TOMATO GEN omic S [29-30 Maggio 2008,Viterbo – Università...

TOMATO GEN omic S [29-30 Maggio 2008,Viterbo – Università della Tuscia] etic

TOMATO GEN omic S [29-30 Maggio 2008,Viterbo – Università della Tuscia] 2etic

Università degli Studi della Tuscia Facoltà di Agraria Dipartimento di Agrobiologia e Agrochimica Proprietà letteraria riservata Grafica e lavoro editoriale: Andrea Mazzucato, Mario A. Pagnotta, Maurizio E. Picarella, Pietro Mosconi Stampa: Centro Stampa Università della Tuscia Finito di stampare nel maggio 2008 Con il contributo di: Università degli Studi della Tuscia (Dipartimento di Agrobiologia e Agrochimica), Società Italiana di Genetica Agraria, Società di Ortoflorofrutticoltura Italiana (Sezione Orticoltura), Barilla S.p.A.

DABAC


“Genetica e genomica del pomodoro:

il contributo italiano”

“Tomato genetics and genomics: the Italian contribution”

Giornata di Studio in occasione del pre-pensionamento del Prof. Gian Piero Soressi

A workshop organized for the retirement of

Prof. Gian Piero Soressi

29-30 maggio 2008

Facoltà di Agraria, Aula del Consiglio - Università degli Studi della Tuscia

The ItalianContributionThe Italian

ContributionThe Italian

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The ItalianContributionThe Italian

ContributionThe Italian

Contribution

SGENTOMATO

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PROGRAMMA Giovedì 29 maggio 14.30 Registrazione 15.00 saluto del Rettore dell’Università degli Studi della Tuscia saluto del Preside della Facoltà di Agraria saluto della Società di Ortoflorofrutticoltura Italiana saluto della Società Italiana di Genetica Agraria Il pomodoro: una prospettiva internazionale Chair: Enrico Porceddu 15.30 Lorenzo Maggioni

BIOVERSITY International “Il gruppo di lavoro Solanaceae di ECPGR: attivita' di cooperazione in Europa per le risorse genetiche”

16.00 Fernando Nuez, Jaime Prohens, María José Díez Università Politecnica Valencia - Spagna “The Institute for the Conservation and Improvement of Agrodiversity (COMAV) input to tomato genetics and breeding”

16.40 Silvana Grandillo CNR IGV, Napoli “International SOL Genome Project: il contributo italiano”

17.10 Coffee break Il pomodoro: una prospettiva italiana I Chair: Carlo Lorenzoni 17.30 Agostino Falavigna

CRA ISPORT Montanaso Lombardo “Contributo di Gian Piero Soressi all'avvio ed alla crescita scientifica a Montanaso Lombardo”

17.50 Andrea Mazzucato Università della Tuscia “Il Codice ‘Soressi’ ”

18.10 Alberto Pardossi Università di Pisa “Incontro tra breeder e agronomi: il caso delle colture idroponiche”

18.30 Roberto Ranieri, Marco Silvestri Barilla S.p.A.


“L'esperienza Barilla nella selezione del pomodoro da industria” 20.30 Cena sociale Venerdì 30 maggio Il pomodoro: una prospettiva italiana II Chair: Mario Pagnotta 9.00 Luigi Ricciardi, Concetta Lotti

Università di BARI “Il gene ol-2 di resistenza all'oidio”

9.30 Gian Piero Soressi Università della Tuscia “Mutagenesi classica e variabilità somaclonale in pomodoro: metodologie sperimentali a confronto e loro sinergie”

10.00 Pierdomenico Perata Scuola Sup.re Sant’Anna, Pisa “Produzione di antociani in Arabidopsis e pomodoro”

10.30 Carlo Rosati ENEA Casaccia, Roma “Carotenoids: key players in tomato fruit ripening”

11.00 Coffee break Il pomodoro: una prospettiva italiana III 11.20 Francesco Cellini, Filomena Carriero

METAPONTO Agrobios “Uso di mutanti per studi di genetica funzionale in pomodoro: l’esperienza di Agrobios dal Blind al TILLING”

11.50 Luigi Monti Università Federico II, Napoli “Il progetto GenoPOM”

12.20 Francesco Salamini Università di Milano “Conclusioni e prospettive”

12.50 Gian Piero Soressi “Chiusura dei lavori”


ABSTRACTS “The Institute for the Conservation and Improvement of Agrodiversity

(COMAV) input to tomato genetics and breeding”

Nuez, F.; Prohens, J.; Díez, M.J. Instituto de Conservación y Mejora de la Agrodiversidad Valenciana (COMAV).

Universidad Politécnica de Valencia, Valencia, Spain. [email protected]. The tomato is the most economically important vegetable crop in Spain. The Institute for the Conservation and Improvement of Valentian Agrodiversity (COMAV) leads the Spanish tomato breeding in the following research areas: genetic resources, breeding for resistance to diseases, breeding for quality, parthenocarpic fruit set. The COMAV holds a collection of almost 4000 accessions of tomato and wild relatives mainly from Spain and LatinAmerica, but also from many other countries. Intensive studies are being conducted on the morphological and molecular diversity of both the cultivated tomato and its wild relatives. Research on resistance to viral diseases is being conducted with Tomato yellow leaf curl virus, Tomato spotted wilt virus, Tomato mosaic virus, Pepino mosaic virus, and Parietaria mosaic virus. Development of resistant cultivars and research on diagnostic methods and molecular aspects is being carried out. Studies on organoleptic and nutritional quality have focused on the search of sources with high content of sugars, acids and antioxidants, and in the development of rapid and accurate methods to measure the content of these constituents. The environmental effect and the genetic control of some of these components have been elucidated. Genetic control of parthenocarpy, as well as physiological and molecular aspects have been also studied. We are comparing the transcriptome of parthenocarpic and non-parthenocarpic fruit set in several tomato lines to identify the main pathways implicated in this process. A strategy to identify the genes responsible for parthenocarpy in these lines has been started using a positional cloning and candidate genes approaches.


“Contribution of Gian Piero Soressi to scientific grow of the Montanaso Lombardo Research Section”

Agostino Falavigna

CRA- Unità di Ricerca per l’Orticoltura di Montanaso Lombardo (LO) Gian Piero Soressi started his scientific activity at the Montanaso Lombardo Section of the Research Institute for Vegetable Crops of Pontecagnano (SA) on May 1968 as a researcher. Since 1973 till October 1983 he was Director of the Section. During this period he started breeding programs on tomato, asparagus, onion, bean and an agronomic project on the utilization in agriculture of waste heat from electric power stations. Gian Piero Soressi addressed each one of his young collaborators to specialize on a single crop and spent a big care to their scientific growth. Although not very common at that time and for the Research Centres of the Ministry of Agriculture, he pushed the collaborators to spend a period abroad in order to be trained in outstanding International Research Institutions. Modern plant breeding may help food branded companies in the daily fight in the market because the new tools offered by genomics allow a very efficient selection of the traits required for some end product innovations.


“The Soressi’s code”

Andrea Mazzucato Dipartimento di Agrobiologia e Agrochimica, Università degli Studi della Tuscia, via

S.C. de Lellis, s.n.c., 01100 Viterbo (Italy) – [email protected] The Soressi’s code does not refer to the genetic code, as would be expected in a workshop of geneticists, nor to professional or ethical codes, as would be expected in celebrating the career of a honest and enthusiastic worker. More simply, we refer to the code that every geneticist has to give to his materials, accessions and generations. Soressi is first a ‘code maker’: since the beginning of his career in the early 60s’ he obtained by mutagenesis a considerable number of mutants that were coded, named, multiplied, described and studied in his research activity. Beside genetic codes, Soressi invented the phenotypic codes, that are a list of symbols used in labels to phenotype more effectively large collections in field trials. Second, Soressi is a ‘code collector’: he did one of the first collections of tomato landraces some forty years ago, when the ‘hybrid era’ was not yet commenced, without forgetting to single out spontaneous mutants segreganting in swards. Then, he collected materials from colleagues having the most important germplasm accessions in the tomato breeding history, as the collection of ‘color genes’ from L. Butler and L.A. Darby, the mutant collections of H. Stubbe and C.R. Rick, the near isogenic lines from J. Philouze, the introgression lines selected by D. Zamir. Beside collecting, Soressi has been studying all his entries as a ‘code user’. For many mutations he deeply studied the expressivity, allelism and potentialities in breeding. In some experiments, he put together sets of dozens of mutations, as a prodrome of the modern ‘-omics’ approach. Finally, he has been and is a ‘code disseminator’, as he distributed the materials he characterized to the scientific community, together with the information and many times with related ideas and suggestions. This sowing of the Soressi’s code has been really extensive and is germinated in many of the advances on tomato genetics and genomics nowadays.


“A joint venture between breeders and agronomist: the case of soilless culture”

Alberto Pardossi

Dipartimento di Biologia delle Piante Agrarie – Università di Pisa Viale delle Piagge, 23 – 56124 Pisa

[email protected] The presentation illustrates the long cooperation between Prof. G.P. Soressi (University of Tuscia) and Prof. A. Pardossi (University of Pisa), which was initiated more than 20 years ago. At first, the work focused on the development of new tomato genotypes suitable for greenhouse soilless culture, in particular for high density ((10-14 plant/m2), short-cycle (1-2 trusses per plant) cropping systems to be carried out without the use of auxin treatment for fruit set. With this goal, a series of experiments were conducted between 1987 and 1992 to investigate the adaptation of different monostem (to-2) and parthenocarpic (pat-2) genotypes to NFT (nutrient film technique). Three main types were identified for their vegetative and reproductive characteristics. It was also found that the monostem genotypes developed normal axillary shoots when grown with high nitrogen content in the nutrient solution. More recently, the cooperation between Proff. Soressi and Pardossi was extended to Prof. R. Izzo research team (also at University of Pisa) to investigate the influence of salinity on fruit qualità of tomato plants grown in closed-loop rockwool culture. In this study, conducted in the framework of PRIN projects, three near-isogenic lines of cv. Gimar differing, with respect to wild-type, in ethylene production and fruit ripening pattern were used: gf (green flesh), nor2 (non-ripening2) and Nr (Never ripe). The same response was observed in all genotypes: the higher salinity growth solution increased dry matter content, total soluble solids (TSS), titratable acidity and ascorbic acid, but reduced crop yield. Our data confirm the lack of any relationship between the larger accumulation of TSS and ethylene production in the fruits of salt-treated plants. Salinity induced an oxidative stress, and the sensitivity to salt treatment was genotype-dependent; for instance, Gimar gf was much less sensitive.


“Barilla Experience in Processing Tomato Selection”

Roberto Ranieri e Marco Silvestri Barilla G. e R. Fratelli – Via Mantova 166 – 43100 Parma

Corresponding author: [email protected] The main areas where branded companies are active to deliver innovative end products are: technology/processing, service/packaging, function/usages and raw materials, which represent a powerful tool of differentiation. Barilla’s approach on a strategic raw material is to know in depth the whole production chain and to understand the critical points to study and to develop research projects. Often, the answer is breeding. Conventional and molecular assisted breeding. Adopting breeding on durum wheat (Triticum turgidum var. durum Desf.) and on processing tomatoes (Solanum lycopersicum L.), Barilla has been able to differentiate some of its products in terms of cost, texture, nutrition and appearance. For tomatoes, a good example is the “Scarpariello case”, a tailor made tomato variety selected by Barilla and Peotec Seed company and used by Barilla to produce outstanding and challenging branded sauces. Industrial sauces business has great potentialities to further develop since, in Italy, for less than 10 % of the pasta dishes industrial sauces are used but usually sauces are still home made. People are still not much confident in adopting industrial sauces because the common idea is that “sauces are all the same”, “all sauces have industrial taste” and “ingredients are not natural”. Sauces containing “Scarpariello” are a challenge to change this perspective. “Scarpariello” variety was selected for outstanding and taste which withstands during the thermal applications to guarantee the best raw material quality. It allows obtaining innovative and distinctive sauces that have been “branded” with the variety name too. In the future, through breeding, it could be possible to differentiate even more among sauces in terms of taste and why not, color and nutritional traits too. The raw material relevance on final product innovation is so distinctive that “Scarpariello story” become the core of the TV advertising campaigns of both pasta and sauces categories. Modern plant breeding may help food branded companies in the daily fight in the market because the new tools offered by genomics allow a very efficient selection of the traits required for some end product innovations.


“The tomato ol-2 gene confers resistance to Oidium neolycopersici”

Ricciardi L.*, Lotti C.**,Pavan S.*, Visser R.***, Bai Y.*** *) Department of Agroforestry, Enviromental Biology and Chemistry, Genetics and Plant Breeding

Unit, University of Bari, Bari 70126, Italy **) Department of Agro-Environmental Science, Chemistry and Crop Protection, University of

Foggia, Via Napoli 25, Foggia 71100, Italy ***) Laboratory of Plant Breeding, Plant Sciences Group, Wageningen University, 6700 AJ

Wageningen, The Netherlands The tomato recessive gene ol-2 (conferring resistance to Oidium neolycopersici) was identified at University of Bari in an accession of Solanum lycopersicum L. var. cerasiforme and localized on chromosome 4 by means of AFLP, RAPD and SCAR markers, using a F2 population obtained crossing the resistant accession, named LC-95 and the susceptible cultivar SuperMarmande. Further studies were carried out in co-operation with Wageningen University in order to study the resistance mechanism of ol-2, observing that ol-2 confers papilla-associated and race-non-specific resistance to tomato powdery mildew caused by O. neolycopersici. Based on homology with recessive genes conferring resistance to powdery mildew in other species, such us barley and Arabidopsis, the tomato candidate gene (named LeMLO1) was identified in Solgenes data base and the comparison of nucleotide sequences of resistant and susceptible genotypes revealed a 19 bp deletion in the resistant parent. In the F2 of a cross between a resistant line bearing ol-2 and the susceptible tomato cultivar (cv.) Moneymaker, a chromosomal fragment disrupting the LeMLO1 coding region co-segregated with resistance. This polymorphism results in a frame shift and thus a truncated non-functional LeMLO1 protein. As both the genetic and the phytopathological characteristics of the ol-2-mediated resistance are reminiscent of powdery mildew immunity conferred by loss-of-function of mlo (mildew resistance locus o) alleles in barley and Arabidopsis, we initiated a candidate gene approach to clone Ol-2. The LeMLO1 was mapped in the chromosomal region harbouring the Ol-2 locus by using both genetic and cytogenetic tools. Complementation experiments, using transgenic tomato lines and virus-induced gene silencing (VIGS) assays, suggested that loss of LeMLO1 function is responsible for powdery mildew resistance conferred by ol-2. Our researches reveal the second example of a natural mlo mutant, suggesting that natural mlo polymorphisms may compensate for the man-made poverty of resistance gene diversity in the gene pool of cultivated species.


“Classical mutagenesis and somaclonal variation in tomato: confronted procedures and synergies”

Gian Piero Soressi

Dipartimento di Agrobiologia e Agrochimica, Università degli Studi della Tuscia, via S.C. de Lellis, s.n.c., 01100 Viterbo (Italy) – [email protected]

This presentation aims, at the same time, to be a sign of gratitude and warning: a deep thanks to my first scientific leaders, proff. Angelo Bianchi and Gian Tommaso Scarascia Mugnozza, scientists in radiogenetics respectively for maize and wheat; a warning for the young researchers under way of tomato genetics and breeding. What I have learnt since the preparation of my graduation thesis on the effects of x-rays on tomato seeds, were the terms “gametic” and “intrasomatic selection”, “true control” and the symbols indicating the different plant progenies issued from the non treated, X0, X0S1, and x-rayed, X1, X2, seeds. Remembering that X1 is analogous to F1, I was surprised in realizing that in most published papers on in vitro induced somaclonal variation, R0 took place of R1 (like X1), the segregating progeny became R1 (instead of R2, like X2), therefore the true R0 like X0 disappeared. Such an inconsistent change, also disapproved by prof. Francesco D’Amato, led to underestimate the importance of the “true control”, so that some pre-existing mutation of the seeds used for obtaining the explant were de facto misinterpreted as somaclones. Unfortunately (or fortunately), I saw with my own eyes some of such mistakes when I was asked to check a few very interesting, putative somaclones. With such a store of experience, when later we also decided to evaluate the efficiency and quality of the in vitro inducible variation, we took care of having a “true control”, by setting up a procedure that permit the recovery of the explant donor seedling. The same procedure was applied in the experiments where we combined chemical with in vitro mutagenesis. In this way we were able to distinguish and separately screen all the plant progenies under study: M0R0 [from non-treated (M0) and non-regenerated (R0) seedlings]; R0M1 (from EMS treated seeds); R1M0 (from cotyledon explant regeneration only); R1M1 (regenerated plantlets from cotyledons of EMS-treated seeds). Through this research we succeeded in obtaining somaclonal variants induced by chemical mutagenesis but recovered via cotyledon regeneration. A proof of this event was an anthocyaninless mutant singled out in a R1M1 progeny. This mutant likely induced by EMS in a cotyledon cell, should have been lost without cotyledon regeneration because it was induced in a tissue that do not give contribution to sexual lineages. No anthocyaninless somaclone has been up to now reported as a result of the sole in vitro cotyledon regeneration.


“Arabidopsis thaliana MYB75/PAP1 Transcription Factor induces Anthocyanin Production in Transgenic Tomato Plants”

Pierdomenico Perata Scuola Superiore S. Anna, Pisa

Tomato (Solanum lycopersicum L.) cv. Micro-Tom plants were transformed with the Arabidopsis thaliana (L.) Heyhn MYB75/PAP1 (PRODUCTION OF ANTHOCYANIN PIGMENT 1) gene. This gene codifies a transcription factor which is involved in anthocyanin production and is modulated by light and sucrose. The transgenic tomato plants expressing AtMYB75 were characterized by a significantly higher anthocyanin production under normal growth conditions in leaves, stems, roots, flowers and, interestingly, in fruits. Anthocyanin accumulation was not widespread but took place in specific groups of cells located in epidermal or cortical regions, or in proximity to vascular bundles. In the vegetative organs of the transgenic plants, where AtMYB75 overexpression was determined, a clear up-regulation of all the main genes involved in flavonoid pathway was also detected. On the contrary, no effect was produced on the expression of the tomato MYB-gene ANT1 (ANTHOCYANIN1) that had previously been identified as a transcriptional regulator of anthocyanin biosynthesis. Additionally, induction of many but not all the structural genes of the biosynthetic pathway was observed in the fruits. The higher basal content of anthocyanins in the leaves of the transgenic plants could be further increased in the presence of high light conditions and contributed to mitigate photobleaching damages under high irradiance. Transformations of Anthocyanin fruit (Aft) and Ailsa Craig tomato genotypes were also performed obtaining similar results. Molecular characterizations of these transgenic plants are in progress.


“Carotenoids: key players in tomato fruit ripening”

Carlo Rosati1, Gianfranco Diretto2, Raffaela Tavazza2, Florence Bouvier3, Bilal Camara3, Einat Bar4, Efraim Lewinsohn4, Nicholas

Schauer5, Alisdair Fernie5, Giovanni Giuliano2 1 ENEA Trisaia, S.S.106 km 419+500, 75026 Rotondella (MT) 2 ENEA Casaccia, via Anguillarese, 00060 S. Maria di Galeria (RM) 3 CNRS, Plant Molecular Biology Institute, 12 rue du Général Zimmer, 67084 Strasbourg, Francia 4 Institute of Plant Sciences, Newe Ya’ar Research Center, Agricultural Research Organization,

P.O. Box 1021, Ramat Yishay, 30095 Israele 5 Max-Planck Institute for Molecular Plant Physiology, Am Muehlenberg 1, 14476 Golm, Germania Carotenoids are C40 plastid-associated molecules which impart nutritional properties and visual appeal to tomato. Due to the strong accumulation of carotenoids in the fruit and the availability of mutants, tomato has long been used as a model species for the carotenoid pathway. The biosynthetic route has been extensively characterized also by metabolic engineering, which have revealed some critical regulatory steps of the carotenoid content in tomato fruits and successfully increased pro-vitamin A and xanthophyll content. Comprehensive and targeted gene expression and metabolite analyses of tomato transformants showed that carotenoid cyclization/hydroxylation is also able to induce global changes in fruit ripening, by modulating fruit transcript levels and metabolic profiles, thereby affecting important traits such as fruit firmness, ABA levels, and the emission of ethylene and volatile compounds.


“Uso di mutanti per studi di genetica funzionale in pomodoro: esperienza di Agrobios dal Blind al TILLING”

Francesco Cellini, Filomena Carriero

Metapontum Agrobios, S.S. Jonica 106, km 448.2, 75010 Metaponto (MT) Italy La disponibilità di mutanti è di straordinaria importanza per il miglioramento genetico di colture in ambito agrario e per la ricerca della funzione di geni di interesse. Le tecnologie per la generazione e lo studio di mutanti hanno registrato notevoli sviluppi, grazie al progresso delle tecniche molecolari ed alle conoscenze acquisite attraverso il sequenziamento del genoma delle piante superiori. La disponibilità di particolari strumenti molecolari ha consentito di mettere a punto, nel corso degli anni, sofisticati approcci di genetica funzionale, quali il transposon tagging eterologo, il gene walking che hanno evidenziato successi e fallimenti in modo variabile. Un esempio dell’evoluzione di tali tecnologie è fornito dall’approccio seguito da Agrobios per l’isolamento e la caratterizzazione del gene blind, e della sua variante allelica torosa,in pomodoro, un caso che dimostra l’importanza di disporre di mutanti ben caratterizzati. Negli ultimi anni è apparsa una nuova potente tecnologia che consente di identificare nuove varianti alleliche di geni, la cui sequenza nucleotidica è nota, e di ottenere nuovi genotipi migliorati nel carattere di interesse. Questa tecnologia, ad oggi applicata su diverse specie, è conosciuta con il nome di TILLING (Targeting Induced Local Lesions In Genomes). Precisamente il TILLING utilizza la mutagenesi chimica (es. EMS) per indurre variabilità (mutazioni) nel genoma di un organismo e tecniche di biologia molecolare (PCR) per individuare mutazioni utili e quindi varianti alleliche di geni di interesse. Attraverso questa tecnologia possono essere selezionati genotipi con nuove caratteristiche agronomico-nutrizionali direttamente trasferibili verso uno sviluppo commerciale. Presso la Metapontum Agrobios, l’applicazione della tecnologia TILLING ha avuto inizio con la produzione, mediante mutagenesi chimica (EMS), di una collezione di mutanti di pomodoro (cv Red Setter) costituita da 6677 Famiglie M2 e 5508 Famiglie M3. Dati sullo sviluppo della piattaforma TILLING relativi sia alla produzione della collezione di mutanti di pomodoro che agli screening molecolari verranno presentati e discussi.


SINOPSI DELL’ATTIVITÀ SCIENTIFICA DEL PROF. GIAN PIERO SORESSI

L’attività scientifica di Gian Piero Soressi, pur mantenendo il comune denominatore dell’utilizzo della variabilità naturale e indotta per il miglioramento genetico di specie orticole (il pomodoro innanzitutto), ha affrontato un gran numero di argomenti, sotto diverse prospettive e, soprattutto, con l’interazione di tantissimi studenti, collaboratori e colleghi. Crediamo che questo prezioso lavoro, a volte rimasto al di fuori della letteratura internazionale più diffusa, meriti una sintesi più puntuale rispetto a una semplice lista di pubblicazioni.

SYNOPSIS OF PROF. GIAN PIERO SORESSI’S

SCIENTIFIC ACTIVITY The research activity of Gian Piero Soressi, although maintaining the common denominator of using natural and induced variation for the genetic improvement of vegetable (mainly tomato) crops, has addressed a myriad of research topics, under a number of perspectives and, most notably, interacting with a great number of students, collaborators and colleagues. We believe that this valuable work, sometimes remained outside of the most diffused international literature, deserves a presentation somehow more detailed than a simple list of publications.


7B-1 mutant Andrea Mazzucato (Università della Tuscia, Viterbo Italy) The conditional male sterile mutant 7B-1, showing sterility in LD and

fertility in SD, has been the object of a collaborative research with Prof. V.K. Sawhney of the University of Saskatoon, Canada. The expressivity of the mutation was characterized in the Italian conditions at Viterbo and sterility was very good in spring-summer (see Figure). However, the recovering of fertility in our autumn conditions was not satisfying (10 seeds per fruit on average). Creating artificial SD in the spring season by covering the plants was proposed as a promising alternative. A possible allelism between 7B-1 and the

class B MADS-box gene DEFICIENS was excluded by mapping experiments (Stamegna 2008).

Aloe arborescens

Chiara Bedini, Antonio Tiezzi (Università della Tuscia, Viterbo Italy) A micropropagation protocol for Aloe arborescens has been developed in order to maximize the multiplication index and to minimize the cycle length. Explants were sterilized in NaOCl and subcultured weekly to

overcome the effects of released polyphenol that, otherwise, caused browning of the cut surface, tissue damage and death. Five substrates having Murashige & Skoog salts as a basis and differing in the type and concentration of auxins and cytokinins were tested. Explants cultured on the substrate containing 1.0 mg l-1 NAA and 2.0 mg l-1 BA showed abundant sprouting from their basement and from axillary meristems, with a mean multiplication index of 3.5. Biological effects of leaf extracts from micropropagated

plants of A. arborescens on proliferation of the murine myeloma cell line P3X were evaluated. Preliminary results confirmed the presence of bioactive molecules and their activity in inhibiting cell proliferation; such

Sterile phenotype of 7B-1 in spring LD conditions (left) and partial recovering of fertility in SD in autumn (right).

Aloe explant with anaxillary sprout obtainedin vitro.


activity was lower than that of analogous leaf extracts from three-year old plants (Bedini et al. submitted).

Anthocyanins Mariagrazia Antonelli, Andrea Mazzucato, Enrico Santangelo (Università della Tuscia, Viterbo Italy) Anthocyanin pigments belong to the class of phenolic compounds named flavonoids and are biosynthesized via the shikimic acid pathway. They provide for red, blu or purple colour in flowers, fruits, stems, roots and leaves, playing an important role in plant reproduction by recruiting pollinators and seed dispersors. Anthocyanins in plants act also in protection against various stresses such as UV irradiation, hight light intensity, ROS, insect and pathogen attacks. In the last years, interest in anthocyanins has intensified because of their health benefits as dietary antioxidants and their pharmacological properties for therapeutic purposes. In tomato, flavonoids are accumulated in a tissue-specific way depending

on the fruit developmental stage. They are mainly synthesized in the epicarp where naringenin, a precursor in anthocyanin biosynthesis, is the main component reaching a peak at the fruit turning stage. Soressi’s collection of tomato germplasm contains accessions of spontaneous or artificially induced anthocyanin mutants, obtained since the 60s, and genetic stocks from the C.M. Rick Tomato Genetics Resource Center. This collection includes monomendelian mutants characterized by a remarkable increment (ag, atv, atvc, atvsc, Aft, Abg), decrement (al) or absence (a, aa, asc, ac, af, ah, aw, bls) of anthocyanins in stem, leaf and fruit.

a bTomato plant originated from the cross atv x Aft and Aft x atvsc: a) “sunblack” with high anthocyanin fruit pigmentation; b) selected plant suitablefor mechanical harvesting with typical Aft pigmentation.


Using spectrophotometric analysis, Santangelo et al. (2002), studied the modifications occuring in tomato fruit pigments during ripening. They found that in the Aft mutant, anthocyanin content decreased during fruit ripening as for chlorophyll. Subsequently, by using fluorescent non-destructive techniques, Lai et al. (2007) analized changes in anthocyanins, chlorophyll, carotenoids in epicarp of tomato during fruit ripening, identifying the main fluorescent bands of these pigments. These results could give important information about the optimal conditions for fruit harvesting using non-destructive techniques. Considering the biological relationship between anthocyanin induction and insect attack, experiments have been carried out in order to verify the effect of tomato leaves with different anthocyanin content (atvsc, a and the control UC82) in the diet of Spodoptera littoralis. Results suggested that high anthocyanin content could give sub-letal effects to the physiology of the larvae (see Spodoptera littoralis). TOMANTHO, a MIUR-National Project (2006), coordinated by Prof. P. Perata from the Sant'Anna School for Advanced Studies of Pisa, involving Soressi’s group as partner, aims to investigate different aspects connected with anthocyanin content in tomato leaves and fruits by using atv, atvc, atvsc and Aft mutants. Objectives of this project are: (i) development of tomato breeding lines with high anthocyanin content in leaf and fruit, (ii) identification of molecular markers linked to the genes atv and Aft, (iii) evaluation of the response of tomato anthocyanin mutants to the phytopathogenic baterium Xanthomonas vesicatoria, (iv) study of phenotipic and agronomic interaction between anthocyanin genes and pigment enhancer genes (see Tomato, “high-pigment genes”). Today, F4 and F5 progenies derived from the atv x Aft and Aft x atvsc crosses have been obtained, and the other objectives are being realized. Within the F4 progenies some double recessive recombinants named “sun black” for their unusual fruit pigmentation, were singled out (see Figure). A previous MIUR-National Project (2005) in collaboration with Prof. M. Badiani at the “Mediterranea” University of Reggio Calabria on the response of some tomato mutants to ozone exposure (see Ozone) showing that the atv mutant seems to be more resistant to O3 than other tomato genotypes, confirming a role of anthocyanins as ROS scavengers.


Artichoke Enrico Santangelo (Università della Tuscia, Viterbo Italy) In the 70’s, through multivariate analysis of 40 relevant phenotypic traits of the artichoke traditional varieties cultivated in Italy, four major affinity groups namely “Spinosi”, “Catanesi”, “Violetti”, “Romaneschi” were recognized. In the framework of the breeding activity developed in our country by R. Tesi and A. Graifenberg from the beginning of the 80’s and relatively more recently by F. Saccardo’s team, rightly enters the enthusiastic, 15 years-selection work of Pietro Papalini. Soressi and Papalini met for the first time in the 60’s for their contribution in trying to solve the problem of the mechanical harvesting of the tomato crop in Italy. The selection work on artichoke by Papalini has been mainly based on

open-hand crosses between clones of different cultivars from the “Romanesco” and “Violetto” groups. By applying such a methodology, he selected a number of clones well characterized for plant vigour, yielding period, shape, size, commercial and organoleptic quality of the heads. To continue his work of open-hand crossing and selection, Papalini created a basic “genetic pool” named

by him “Club della Biodiversità” formed by his best selected clones (Gigante, Moro di Corneto, Pacific, Marengo, Chiarostar, Etrusco, Apollo, Exploter, Roma99), integrated afterwards with new entries endowed with special traits required by fresh market or industry. At this point Papalini’s experience met the Soressi’s one as geneticist and breeder (Soressi 2003). From this cooperation, research on artichoke genetics continued in the frame of the experimental PRAL program “Artichoke Varieties Innovation and Cultivation Planning” supported by the Lazio Region and coordinated by the phytopathologist Prof. Paolo Magro. The activity carried out in two years of research included the selection of new F1-open clones and a bio-agronomic field trial in which the traditional cv. Castellammare, new clones of Romanesco type (Grato1, Grato2, C3), cv. Apollo (Vitroplant), Exploter and Giove (both from Papalini) were compared. This field trial proved the earliness of C3 compared with the other clones. Each clone showed distinct plant habit and head features (size, shape and colour nuance). Apollo exhibited a peculiar minor leaf insertion angle possibly responsible of its

Plant of the cv Giove showing the nearlyhomogeneous ripening of 1st and 2nd orderheads.


higher tolerance to cold and to other unfavourable climatic events. Giove reached the maximum concentration of head emission permitting to harvest all the 1st and 2nd rank heads only 9 days after the main head (see Figure); thus it appeared particularly suitable for artichoke production destined either to fresh market or to the industry. The selection activity carried out on artichoke appears promising even considering the difficulties in putting together in few clones all the desired traits. The “gene-pool” established with the various types of the artichoke clones enriched by the best F1-open clones selected by Papalini in 2005 is a valid premise for selecting promising F1-clone combinations among the new F1-open plants from the open-hand crosses obtained in 2006 and observed during the 2008, first year of growth.

Asparagus Agostino Falavigna (CRA – Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) In Italy the unique breeding program on asparagus, promoted and started in 1974 by Soressi, at that time head of the ISPORT Section (Soressi and Falavigna 1979; Falavigna and Soressi 1981a), is still working at CRA – Research Unit for Vegetable Crops of Montanaso Lombardo. Such a program has been constantly financed by the Emilia Romagna Region (through CRPV in Cesena), Veneto Region (through Veneto Agricoltura) and the Italian Ministry of Agriculture in the sub-project “Asparagus and Onion” (Soressi 1986b; 1988). Since the conventional breeding methods based on selfing cannot be easily applied to asparagus, a dioecious and perennial species, the research activity progressed through the “Diego” F1 hybrid (Falavigna et al. 1982, XXVI SIGA Annual Congress), obtained by crossing two heterozygous plants (selected few years for their spears yield and quality), until to the double haploid (DH) clones; in both cases the parent clones were micropropagated (Falavigna and Soressi 1981b; Falavigna et al. 1984a). To obtain DH clones, in vitro anther culture was sistematically applied (Fig. 1, 2). The “super-male” androgenetic DH clones originate all-male hybrids (AMH) of two kinds: two-ways AMH, when both parents are DH clones; three-ways AMH, when the seed parent is an heterozygous clone. All DH clones were evaluated in fields naturally infested by Fusarium spp. (the most important soil born disease of asparagus) and selected for plant survival ratio, fertility and morphological traits. During thirty years of work, about 300,000 anthers of Italian asparagus landraces and hybrids from abroad were cultured. The in vitro culture technique on average yielded 1 androgenetic clone out of 100 anthers and


the percentage varied from 0.1 to 30 according to the anther donor genotype. Out of 3,000 androgenetic clones obtained, 58% were DH and the female/male ratio was near 1/1. The selection pressure over the DH clones was about 95%; therefore 103 and 100 respectively female and male clones were selected and systematically utilized as parents of all male F1 hybrids (Falavigna et al. 1983; 1984b).

Seedling growth analysis was carried out by comparing 3 new AMHs with commercial hybrids and local populations (Stancanelli 1989; Stancanelli et al. 1990). Moreover, a collection of asparagus wild species germplasm in Sicily, previous botanical classification, was tested for seed germination and in vitro vegetative propagation (Fig. 3; Venezia 1991; Venezia et al. 1993). Out of 350 F1 all male hybrids tested so far, ten have been included in the National Variety Catalogue and four (Eros, Marte, Ercole, Zeno, Fig. 4) are grown at present on almost the whole green

asparagus cultivation area and on a great part of that cultivated for white spear production in Northern Italy (about 2,000 ha). Besides, the AMH Italo is risen in Southern regions (Falavigna 2007). Further improvements for spears yield and quality, plant disease resistance and salt tolerance are expected

on the basis of multifaceted investigations under way by the Falavigna research group (Falavigna et al. 2008).

Fig.1 Callus proliferatingfrom a pollen grain inasparagus anther.

Fig.2 Androgenetic calli regeneratingasparagus stems ready formicropropagation.

Fig.3 Asparagusspears of the twomost important F1all-male hybrids.

Fig.4 A plant of thewild species A.maritimus.


Association genetics Andrea Mazzucato, Maurizio Enea Picarella (Università della Tuscia, Viterbo Italy) The characterisation of very diversified materials with molecular markers offers a unique opportunity to define significant marker-trait associations of biological and agronomic interest. In this research, carried out in collaborations with colleagues of the university of Ancona (prof. Papa and Nanni) and Perugia (prof. Negri and Veronesi), 50 tomato landraces (mainly collected in central Italy) nine vintage and modern cultivars, and two wild outgroups were characterised for 15 morpho-physiological traits and 29 simple sequence repeat (SSR) loci. The markers were selected to include a group of loci in regions harbouring reported quantitative trait loci (QTLs) that affect fruit size and/or shape (Q-SSRs) and a group of markers that have not been mapped or shown to have a priori known linkage (NQ-SSRs). Compared to the low molecular polymorphism reported in tomato modern cultivars, our data reveal a high level of molecular diversity in landraces. Such diversity has allowed the inference of the existence of a genetic structure that was factored into the association analysis. As the proportion of significant associations is higher between the Q-SSR subset of markers and the subset of traits related to fruit size and shape than for all of the other combinations, we concluded that this approach is valid for establishing true-positive marker-trait relationships in tomato (Mazzucato et al. 2008a).

atroviolaceum (atv) see Anthocyanins

Auxin

see Reporter gene; parthenocarpic fruit (pat); Tomato, leaf peroxidases

Bean, biotechnological approach

Emilio Mendoza de Gyves, Enrico Santangelo (Università della Tuscia, Viterbo Italy) Andrea Allavena (Res. Unit for Floriculture and Ornamental Species, Sanremo Italy) Ali Akbar Habashy (Agricultural Biotechnology Research Institute, Karaj Iran) Prof. Soressi’s activity on in vitro tissue culture, somatic embryogenesis and genetic transformation of bean (Phaseolus spp.) begun in the 80’s at the Research Institute for Vegetable Crops of Montanaso Lombardo (Lodi) (Castelnuovo 1984; Angelini Rota 1987). During the 90’s the efforts continued in the “In vitro Culture Laboratory” at DABAC (Tuscia


University). At the end of the 90’s, the group participated to an European Concerted Action (see Phaselieu Concerted Action) and to the Phaseolus Italian Network (PIN). Studies were conducted to determine the effect of growth regulators on the micropropagation aptitude of several genotypes of common (P. vulgaris) and running (P. coccineus) bean coming from different countries (Mexico, Peru, Iran, Italy, Colombia, etc.). Axillary and apical bud explants were cultured to ascertain any different response to in vitro culture conditions. Multiple buds production and shoot elongation, with different level of response, were obtained from all genotypes: BAP was able to give a satisfactory percentage of multiplication, but, at all concentrations used, it induced abundant callus at the base of the explants therefore reducing adventitious bud formation in all cultivars (Allavena et al. 1994; Mendoza de Gyves 1995). A protocol for efficient plant propagation via axillary bud explants was proposed for cv. Montecarlo and with some adjustments was successfully utilized for other genotypes such as Giulia, Pinto, Clio, etc., including the running bean, which is more amenable to tissue culture than common bean (Mendoza de Gyves 1995). Mature seeds of the Italian cvs Montecarlo (P. vulgaris) and Streamline-770 (P. coccineus) were utilized for further studies. The joint axes between cotyledons and embryos, a tissue rich of meristematic cells with high aptitude to differentiate, were used. Abundant shoots were produced from the explants in presence of different concentration of BA and/or TDZ. The results showed that the absence or presence of cotyledons (a rich source of nourishment and growth stimulation) is a determining factor to affect individual tendencies and characteristics about the number of buds (Mendoza de Gyves 1995). A first attempt of regeneration from somatic tissues was made starting from leaf-derived calli induced on a medium containing 2,4-D. After transferring on liquid secondary medium with 2,4-D replaced by NAA, KIN, ABA and GA3 the embryo-like structures reached the heart stage but did not became plantlets (Allavena et al. 1994). Focusing the efforts on P. coccineus, a species that gives fertile offspring when crossed to P. vulgaris, direct somatic embryogenesis and organogenesis mediated by small glossy calluses were obtained from immature cotyledon explants on a modified half-strength MS medium containing various concentrations of growth regulators. The availability of an efficient protocol for plant regeneration from somatic tissues and the susceptibility of the species or cultivar to infection by Agrobacterium are required for successful transformation (Parson 2000). A compatible reaction between Phaseolus and Agrobacterium was demonstrated in our works. The presence of a detectable level of nptII


activity in calli derived from several bean genotypes confirmed that kanamycin resistance can be utilized as selectable marker in experiments aimed to obtain transgenic plants (Allavena et al. 1994). A home-made high-velocity microprojectiles bombardment device was constructed and set up in Soressi’s lab. The efficiency of biolistic gene

delivery was tested in cotyledons and embryo axes of immature and mature bean seeds using plasmid pBI121, carrying the ß-glucuronidase (GUS) gene under the control of the CaMV 35S promoter. In P. coccineus, 400±178 units transiently expressing the GUS gene were found per shot. Recovery of transgenic plants was attempted by bombarding the apical meristem of both P. coccineus and P.

vulgaris embryo axes to avoid cell re-differentiation from somatic tissues. On average, 60% of the meristems showed at least one GUS expressing unit after three consecutive bombardments on the same target (see Figure). Chimaeric shoots expressing the GUS gene were sporadically found by histochemical assay (Allavena et al. 1994; Mendoza de Gyves 1995).

(a) Transient GUS expression on Phaseolusembryos after two days of bombardmentusing tungsten microparticles covered withDNA. (b) Stable expression on Phaseolusshoots after one month of infection with Agrobacterium. In both images, arrows pointto blue spots showing the expression of GUS(Mendoza de Gyves 1995).


Bean breeding Andrea Allavena (CRA - Research Unit for Floriculture and Ornamental Species, Sanremo, Imperia Italy) Bruno Campion (CRA - Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) At the beginning of 70th Prof. Salamini together with Prof. Soressi set up a robust breeding project mostly committed to develop bean cultivars resistant to bean common mosaic virus (BCMV) and suitable for both direct

consumption and processing (canning and freezing). Two were the procedures adopted: crossing and selection; seed mutagenesis induction. Along the first way with Fadda and Allavena collaboration, the I gene conferring hypersensitivity resistance to BCMV was transferred from USA cv. Royal Red Kidney (RRK) to Borlotto-type local varieties. Mary, Bea, Inge were the first BCMV resistant varieties released in Italy (Salamini et al. 1978). With the second approach, the RRK red coloured dry seeds were treated with EMS

mutagen and M2 populations screened for seedling, plant and seed mutations (Fig. 1; Motto et al. 1975). One of these white seed-coat mutants, after 1 backcross to RRK, led to release BCMV resistant, Lisa P71 (Motto et al. 1979b). By inter-crossing the best BCMV resistant Borlotto bush breeding lines and subsequent selection, new BCMV resistant varieties improved for agronomic and market characters were obtained: Giulia, Lena and Minia (Fig. 2; Allavena et al. 1982a). Then, a new series of BCMV resistant varieties were developed: white seeded Montebianco and Niveo, by inter-varietal crosses (Allavena et al. 1985a); Montalbano, marble white seeded and Mogano, beige seeded, by EMS treatment (Allavena et al. 1985b). A new breeding programme, started in 1975, was focused on the introgression of the genetic tolerance to Pseudomonas phaseolicola (from the PI150414 accession) into the best BCMV resistant lines issued from the previous programmes. Highly tolerant offsprings were

Fig.1 Seed-coat colourmutations from RRKred seeds undergone toEMS treatment.

Fig.2 Borlotto-type bushplant at waxy pod maturityof the BCMV resistant cv.“Lena”, suitable tomechanical harvesting.


backcrossed to BCMV resistant lines and selected for resistance to both diseases following artificial infection with both the pathogens. Monviso and

Morena were released as the best performing bush breeding lines for fresh and dry grain products (Allavena and Soressi 1982; Allavena et al. 1982b). Other halo blight partial resistant lines were Montecarlo and Monterosa (Allavena et al. 1989). Grazia and Patrizia (Fig. 3) were new bush, BCMV resistant, yellow-podded cultivars, developed from [Meraviglia di Venezia x (RRK and Kinghorn wax)] crosses’ breeding lines (Allavena et al. 1986). The breeding work started by Prof. Salamini and Soressi in 1971 for dwarfing P. coccineus through inter-specific cross with P. vulgaris went on with Campion, leading

to the development and release of the white seeded Venere and the mottled seeded Alarico cultivars (Figg. 4 and 5; Campion et al. 1981; Campion 1995). Moreover, the Salamini and Soressi research group studied seed size

inheritance in a cross of wild and cultivated common bean (Motto et al. 1979b), performed growth analysis in a reduced leaf bean mutant (Motto et al. 1979a), contributed to understand sucrose metabolism (Odoardi et al. 1976a) and detected multiple forms of phosphoesoisomerase enzyme in bean seed during its development in the pod (Odoardi et

al. 1976b). Carbohydrates metabolism was also investigated in smooth and wrinkled pea developing seed (Odoardi et al. 1976c).

bitter fruit (bt)

see Glycoalkaloids blind (bl)

see Monostem for tomato processing

Bowman Birk gene see Helicoverpa armigera

Fig.3 Extremely bushyplant with yellow podsof the BCMV resistantcv. Patrizia.

Fig.4 Seeds of the cv.Venere.

Fig.5 Seeds of the cv. Alarico.


brown seed (bs) see Tomato, genetic markers

Bruschetta see Tomato, “fruit colour genes”

Cladosporium fulvum-2 (Cf-2)

see “Lesion mimic” mutants Cystein proteinase inhibitor

see Helicoverpa armigera DDRT-PCR

Giuseppe Testa, Andrea Mazzucato (Università della Tuscia, Viterbo Italy) Differential display RT-PCR has been adopted to study differencies in gene expression in two experimental systems. In garlic, explants have been

compared before and after the induction of in vitro proliferation in order to identify genes involved in adventitious sprouting. The results have been included in the PhD thesis of Dr. Paula Bima (Bima 1997). In tomato, RNA populations extracted from ovaries of the parthenocarpic fruit (pat) tomato mutant and from the corresponding near isogenic wild-type were compared. The sequencing and the characterization of three transcripts that were differentially displayed in pat mutant ovaries compared to the wild-type were described in detail (Testa et al. 2002). The protein predicted by the full-

length cDNA reconstructed from Clone 66 showed homology to peptides encoded by self-incompatibility alleles of Papaver rhoeas and had a strict ovary specific expression. The up-regulation seen at fruit set was an event specific to the expression of parthenocarpy, because it was also found in ovaries of a parthenocarpic fruit-2 line, a mutation which is not allelic to pat. The putative polypeptide deduced from the cDNA reconstructed from Clone 72 showed high homology with plant H2A histones and preferential transcription in ovaries. A third fragment (Clone 91) was identical to the 3’ region of GAD3, a tomato mRNA sharing similarities to short-chain alcohol dehydrogenases. RT-PCR-based expression analysis revealed that this gene is preferentially

Subepidermic placentalexpression of GAD3 inthe tomato ovary (Testaet al. 2002).


transcribed in ovaries, sepals and leaves. The tenet that this enzyme family has important functions in developmental pathways controlled by hormones, and in particular gibberellins, is supported by its increased transcription at the time of fruit set, by its positive response to treatments with gibberellin A3, and by its expression localized in the mitotic cell layers of the placenta (see Figure).

Dieffenbachia spp. Enrico Santangelo (Università della Tuscia, Viterbo Italy) Claudio Carrai (ARSIA, Pisa Italy) Dieffenbachia is an ornamental species of relevant importance grown in Europe and Usa. A five-years research program was set up in order to define the heritable basis of leaf variegation and create new varieties by means of intervarietal and interspecific crosses.

To make available a crossing procedure efficient for genetic investigations and breeding programmes in Dieffenbachia spp., some experiments were carried out. The degree of receptivity of the stigmas and the fertilizing capacity of the pollen were studied in relation with different flower phenological stages. Attention was also given to seed extraction and conditions favourable to seed germination and seedling growth. The crosses involved the commercial cultivars ‘Alix’, ‘Camilla’, ‘ Candida’,

‘Compacta’, ‘Jeanette’, ‘Katharina’, ‘Marianne’, ‘Nelly’, ‘Tropic white’, ‘Veerle’. On some of these cultivars both selfing and sib-cross were accomplished. Fruit setting, seed germination and albino seedling segregation were evaluated (Carrai et al. 1990; see Figure). Usually the environmental conditions of the greenhouse where the Dieffenbachia is grown hamper the flowering, that remain sporadic, poor and progressive. Flowering has no value in the commercial production of plants, but it is of basic importance for breeding programmes. With the purpose of increasing the number of flowers and synchronizing their emission, the effect of GA3 treatments was studied (Fioravanti et al. 1991). Two different periods of GA3 application (December and February) and four cultivars with different ploidy levels (8n and 4n) were tested. All the cultivars responded positively

Dieffenbachia ripening fruits(Carrai et al. 1990).


to the treatment and showed an interaction between treatment period and genotype for the number of flowers per plant and their concentration of emission. The number of days from GA3 application to flower appearance was lower in December than in February. The ploidy level of the genotype seemed to influence both cross compatibility and response to GA3 treatments (Carrai et al. 1993). The number of flowers induced by GA3 was dependent on plant age, vigour and branching.

easy peeling (ep) see “Fruit skin” mutants

Ethylene

Enrico Santangelo, Valentina Fonzo, Mariagrazia Antonelli, Maurizio Enea Picarella (Università della Tuscia, Viterbo Italy) Although considered as the “ripening hormone”, ethylene plays a role in many metabolic processes. The tomato “never ripe” group includes a number of mutants with a direct or inderect impact on the synthesis and/or perception of ethylene, displaying a phenotype with delayed and incomplete ripening (Soressi 1975). Soressi’s activity has been concentrated on the study of the tomato “never ripe” mutants Nr, rin and nor2, encompassing

also aspects of plant physiology other than fruit ripening. Studies of such mutants have been developed during the 70’s (see “Never ripe” mutants). The availability of such ethylene-related genes in the same or

different backgrounds has extended concerns towards other physiological aspects as seed germination and the role of ethylene in response to some abiotic stresses. In Nr, rin and nor2, a more prompt and higher germination percentage associated with a lower level of ethylene emission per seed compared to the WT was observed (Lai et al. 2002, XLVI SIGA Annual Congress; see Figure). By taking into account that 20 year-old seeds of these ethylene mutants are still able to germinate, it can be speculated that in these genotypes ethylene accumulation during seed senescence occurs slowly and some processes, in which other hormones like auxin are involved, are somehow altered during germination.

nor2 rin WT

Germination of 5-year-old seeds from nor2, rin and WT 3 days after sowing.


Considering the role of ethylene as a signal molecule, the study of its effect following treatments with abiotic constrains like high temperature, luminosity, ozone and salt has been undertaken. The presence of Nr, rin and nor2 genes has altered and modulated the leaf necrotic lesions occurring in a tomato line displaying acropetalous autonecrosis at high temperature and increasing luminosity (Fonzo et al. 2003, XLVI Annual SIGA Congress; see “Lesion mimic” mutants). When Nr and rin mutants have been exposed to a single, relatively high dose of O3, an abiotic elicitor in plants, the results suggested that the relationship between ethylene emission and the severity of O3-induced foliar symptoms might be less straightforward than expected (Antonelli et al. 2005, XV Meeting of the Eucarpia Tomato Working Group; see Ozone). Finally, research on salt tolerance started in the first 90’s, has recently involved the use of the nor mutant with the aim of analysing the interaction between ethylene and abscisic acid (Picarella et al. 2007; see Salt tolerance).

Fagiolo Badda see Landraces, bean

Fagiolo del Purgatorio

see Landraces, bean

Fen gene Mariagrazia Antonelli, Enrico Santangelo, Giorgio Mariano Balestra, Leonardo Varvaro, Margherita De Biasi, Stefania Astolfi (Università della Tuscia, Viterbo Italy) In tomato, the resistance to race 0 of the phytopathogenic bacterium

Pseudomonas syringae pv tomato (Pst) is governed by Pto, a member of a clustered gene family that also includes the Fen gene that confers sensitivity to the organophosphorous insecticide Fenthion (see Figure). Therefore, tomato cultivars that contain Pto also exhibit a HR-like response to the insecticide. Other components of the Pto resistance signaling pathway are the Prf and PtiI genes. In vitro regeneration on a medium containing increasing doses of the insecticide was employed to obtain regenerants resistant to Fenthion. Cotyledon explants of the hybrid derived from the cross Rimone (resistant to Pst

Necroses on leaves oftomato cv. Rimoneafter Fenthion spray.


and sensitive to Fenthion) and Riogrande (resistant to Fenthion and sensitive to Pst) were used. Santangelo et al. (1998) reported that the methods allowed to induce variation in the Pto-Fen-Prf genic complex and the selection of R2 plants with a different degree of tolerance to both the agents (bacterium and/or insecticide) (Santangelo et al. 2000). In order to explore the functional relationships among the expression of the Pto/Fen gene family and the GOX transgene (see Glucose oxidase, GOX), four tomato NILs (Riogrande and Rimone and their respective GOX-transformants, RC332 and RC131) were exposed to Fenthion, evaluating the production of H2O2 and the activities of ROS-scavenging enzymes in foliar tissue. Accumulation of H2O2 and transient enhancement of ROS-related enzymatic activities could together confirm that Fenthion does elicit an oxidative burst in the tissues of the tested tomato lines driven by either Fen gene and GOX transgene. The Fen gene conferred sensitivity to Fenthion, regardless of the expression of GOX. A prolonged accumulation of H2O2 was observed in the leaves of Rimone and RC131, which was instead transient in Riogrande and in RC332. In all the tomato lines, exposure to Fenthion induced rapid transient changes in the activities of most enzymes, except for peroxidase activity in the leaves of Rimone and RC131, that steadily increased until the end of the sampling period. It is suggested that the sensitivity of Rimone to Fenthion might be due to the sustained activity of a H2O2-forming peroxidase (De Biasi et al. 2003). In a study involving the tomato autonecrotic mutant V20368 (see “Lesion mimic” mutants), treatments with Fenthion were performed to determine whether the insecticide induced foliar lesions. Though the V20368 line possesses the recessive pto and fen alleles, a necrotic phenotype mimiking autonecrosis was observed after Fenthion treatment evidencing a lower threshold of stress perception of this tomato line (Santangelo et al. 2003a). The appearence of necroses on V20368 after Fenthion treatment was confirmed by trypan blu staining followed by stereomicroscopic observations (Antonelli et al. 2006, XIII SIPAV Annual Meeting).

“Fruit skin” mutants Antonella Portesi (Università Cattolica Sacro Cuore, Piacenza Italy – presently different address) Piero Frangi (Fondazione Minoprio, Como Italy) Giorgio Chioccia (Università della Tuscia, Viterbo Italy) The tomato fruit, highly variable in weight and shape, according to genotype and growth conditions, is endowed with a protective skin or exocarp. Beneath this outer epidermal layer, there are 2 to 4 layers of hypodermal cells with thicker walls and collenchyma-like thickenings. A


layer of cutin is upon the thin (4-10 μm) cuticle, that covers the epidermal cells. Cutinization extends into the radial walls of epidermal cells and may also involve the hypodermal ones. The trichomes developed from epidermis disappear at fruit maturity. Heavy cutinization contributes to the fruit cracking resistance. Genotypic differences in pericarp and skin anatomy exist and some of them are of possible interest in tomato breeding. The mutants up to date described are: sticky peel (pe), peach (p), easy peeling (ep), waxy fruit (pe-2) and non-ripening-3 (nor3). pe gives crack-resistant fruits but with a shiny, extremely elastic and thin skin, possibly lacking of hypodermis. pe-2, mimicking pe, appeared in cv. Manapal. The p fruit has a dull, opaque surface with increased hairiness when still immature. The ep phenotype is caused by the disintegration of parenchyma cells with intercellular air spaces below the hypodermis, thus permitting an ‘easy peeling’ of the fruit. This mutant is of possible use in tomato breeding for domestic or industrial peeled products. nor3 is a spontaneous mutation appeared in the Italian “Tondo liscio di Pescara” landrace with colorless and dull skin fruit, slow in ripening (Soressi 1975). Studies on the genetic relationships among pe, pe-2, p and nor3 revealed that pe-2 is allelic to pe and that nor3 is allelic to p. As a consequence and according to the TGC rules the new symbols are pe2 instead of pe-2 and p2 instead of nor3 (Portesi

1987; Portesi et al. 1987). Observations by scanning electron microscope and histological analyses have evidenced a clear-cut difference in the cuticle of the mutants: it results practically absent in pe2, thicker and likely rugose (dull) in p2, thick and smooth on the WT cv. Gimar (Fig. 1). Such differences are recognizable from an early stage of fruit development. Likely, quali-quantitative variations in cuticle waxes produced by the

epidermal fruit cells are involved. Another feature peculiar to p2 mutant is an early abscission of the fruit at the pedicel joint (Fig. 2) in relation with water availability. In addition the p2 fruit is resistant to cracking disorder

Fig.2 Gimar fruit with p2

phenotype, showing precociousjoint-pedicel abscission.

a dcb

Fig.1 Cuticole morphology of pe2 and p2

mutant fruits compared with cv. GimarWT: breaker (a) and red ripe (b) stagesof the p2 mutant; red ripe stage of pe2

mutant (c) and WT (d) (Portesi 1987).


on-vine and slowly shrivels during storage due to a higher permeability of its skin to gases (CO2, O2, ethylene) and water. The p phenotype can also ave some interests in tomato breeding by reducing fruit water content, with lower pathogenic fungi attacks and also resulting in cracking resistance. To avoid an early abscission of the fruit, it would be valuable to study the combination of p2 with jointless mutants (j, j-2), thus improving the performance against the mechanical harvesting shakes. The lower water content of p mature fruits, besides requiring less heating power in concentrating tomato puree, makes them particularly suitable for preparing the Viterbese “Panzanella” food, made by spreading tomato red pulp onto a slice of dry bread, previously soaked in fresh water, and pouring on it a spill of extra-virgin olive oil.

Genetics and breeding Andrea Mazzucato (Università della Tuscia, Viterbo Italy) Despite his preference for following lab research and field trials, Soressi was also engaged in writing contributions of general and/or didactic interest, in the field of the teaching of genetics (Bianchi et al. 1973), plant breeding (Soressi 1986a; Blanco and Soressi 1988), genetic resources conservation (Maggioni and Soressi 1992) and agronomy (Soressi et al. 1981; Caporali and Soressi 1993).

Gibberellins Andrea Mazzucato, Irene Olimpieri, Francesca Siligato (Università della Tuscia, Viterbo Italy) The role of gibberellins (GAs) in the development of male and female tomato floral organs has been addressed in the study of the parthenocarpic fruit (pat) phenotype (see parthenocarpic fruit). Whereas early research showed that GA was able to rescue the anther phenotype but not parthenocarpy in the mutant (Mapelli et al. 1979), a revision of the sensivity of the mutation showed that gibberellic acid (GA3) was only effective in restoring carpelloid anthers to the wild-type phenotype (Mazzucato et al. 1999). Further reports indicated novel pat phenotypes that parallel those reported in plants repeatedly treated with gibberellic acid or having a GA-constitutive response. The retained sensitivity to the GA-biosynthesis inhibitor paclobutrazol revealed that this condition is dependent on GA biosynthesis. Expression analysis of genes encoding key enzymes involved in GA biosynthesis shows that in normal tomato ovaries the GA20ox1


transcript is in low copy number before anthesis and only pollination and fertilization increase its transcription levels and, thus, GA biosynthesis. In

the unpollinated ovaries of the pat mutant, this mechanism is de-regulated and GA20ox1 is constitutively expressed, indicating that a high GA concentration could play a part in the parthenocarpic phenotype. The levels of endogenous GAs measured in the floral organs of the pat mutant support such a hypothesis. Collectively the data indicated that transcriptional regulation of GA20ox1 mediates pollination-induced fruit set in tomato and that parthenocarpy in pat results from the mis-regulation of this mechanism. As genes involved in the control of GA

synthesis (LeT6, LeT12 and LeCUC2) and response (SPY) are also altered in the pat ovary, it is suggested that the pat mutation affects a regulatory gene located upstream of the control of fruit set exerted by GAs (Olimpieri et al. 2007).

Glycoalkaloids Andrea Mazzucato (Università della Tuscia, Viterbo Italy) The tomato, as almost all species of the Solanaceae family, produces glycoalkaloids (GLA, alpha-tomatine and dyhydrotomatine), secondary plant metabolites that show toxicity against vertebrates. The desirability of such metabolites in the tomato fruit is still a debated question; if GLAs are of concern for consumers because of their general cellular toxicity to vertebrates, they have proven to exert toxicity towards bacteria, fungi, viruses and even arthropods, thereby serving as a potential source of resistance to plant pathogens and pests. Recently, it was demonstrated that assumption of alpha-tomatine in the mice diet is 20-fold less toxic than that of other glyco-alkaloids and that the molecule also shows antitumoral and anti cholesterol action. Genetic knowledge on the control of GLA levels is fragmentary. In wild cherry tomatoes, two accessions with bitter ripe fruits were found in some Peruvian districts and the bitterness of their fruits was related to very high content of GLAs (Rick et al. 1994). In Italy, a variant with highly bitter fruit was discovered in a survey for somaclonal variation in the background of cv. UC82 (Cunico, 1991; GP Soressi, unpublished results). The trait, that was given the provisional name of bitter fruit (bf), proved to be recessive and monogenically controlled

Polycotiledony of a patseedling, a phenotypeconnected with alteredGA synthesis and/or GAresponse regulation.


(Cunico 1991). Although the causative relationship between the bitter taste and a high tomatine content was not definitely proved, the phenotype strongly recalls an impairment in GLA composition and represents a valuable material being nearly isogenic to UC82.

Glucose oxidase, GOX Riccardo Caccia, Stefania Astolfi (Università della Tuscia, Viterbo Italy) Transgenic tomato (Solanum lycopersicum L.) plants containing the Gox gene encoding glucose oxidase from Aspergillus niger were obtained in two near isogenic lines (NILs) except for the Pto resistance gene (Riogrande +/+ and Rimone Pto/Pto). T3 transgenic plants possessing a single Gox gene copy were tested for resistance to Pseudomonas syringae pv. tomato

to ascertain the role of H2O2 in plant pathogenesis response (Caccia et al. 1996, XL SIGA Annual Congress). Reactive oxygen species such as H2O2 are generated following pathogen recognition, and they act as both cellular signalling molecules and direct antimicrobial agents. The Gox transgene was regularly inherited and expressed in the tomato plants. The level of GOX activity was one- to five-fold higher than in the non-transgenic WT (see Figure). The H2O2-generating, trans-Gox plants

proved to confer to the susceptible NIL the capacity to respond actively to P. s. pv. tomato by reducing the number of viable bacterial cells in the inoculated leaves and to limit the disease symptoms in terms of number and size of leaf necrotic spots (Caccia 1989; Caccia et al. 1999; Balestra et al. 2000). To deeply explore the activation modes and timing of the plant enzymatic systems involved in biotic and abiotic stress responses, the effects of the Gox gene were studied by considering the interaction between the Pto and Fen genes in the abovementioned NILs, taking into consideration that the Rimone line is susceptible to the Fenthion molecule based insecticide (see Fen gene). These transgenic NILs were also investigated for assessing the damage level caused by polyphagous spider mites (see Tetranychus urticae).

green flesh (gf)

see Tomato, “high-pigment genes”

Different blue color intensityof leaf disks from transgenicGox left) and control (right)plants following anhystochemical assay.


Helicoverpa armigera Stefano Speranza, Riccardo Caccia, Valentina Fonzo, Elia Poerio, Claudio Pucci (Università della Tuscia, Viterbo Italy) Helicoverpa armigera (Hübner) is a cosmopolite Lepidoptera Noctuidae able to injury several plant species among which tomato. The young larva is

yellow with black head and, at the mature stage, it becomes green-yellowish with typical lateral light stripes. It accomplishes its

life cycle in about a month, but in Central-Southern Italy, it performs 2-4 generations per year, over wintering as pupa in the soil (Fig. 1). In the last years, this noctuid has provoked infestations to tomato of so high degree to become a key pest in Central Italy and in several other world regions (Speranza 2001). On the tomato plant, the larvae start eating the leaves, then feed themselves with flower buds and berries, by causing their dropping to the ground or rotting (Fig. 2). At harvest (August in Italy), non marketable fruits exhibiting holes and tunnels without or with eating larvae, can be observed. Proteinase inhibitors are able to interfere in several animal digesting processes and to horn on their growing

(Brodway and Duffey 1986; Ryan 1990). Thus, plant proteinaceous proteinase inhibitors have potential for increasing resistance of crop to insect pests. With the aim of obtaining tomato transgenic plants resistant to

Fig.1 Life cycle of Helicoverpa armigera (Hüb.).

Fig.2 Tomatoes damaged by H. armigera.

Fig.3 Inhibition of cystein proteinaseactivity in the raw extract of H. armigeralarvae according with increasing quantityof proteins extracted by BG24 (—) andRIG (—) leaves.


H. armigera larvae, we transformed - the cultivar Riogrande with the gene KTI3 (coding for a soybean Kunitz Inhibitor) and the cultivar UC-82 with the gene Pi-IV (coding for a soybean Bowman-Birk Inhibitor). Leaf extracts of transgenic plants contained significant levels of inhibitory activities towards both bovine pancreatic trypsin and insect trypsin-like enzyme, on average 187- and 5-fold higher-than controls, respectively. The trypsin inhibiting activity was found stable at different plant phenological phases in fruits and leaves; this activity resulted stable for 24 h in detached leaf discs used to feed insects. Both inhibiting activities were able to contrast development of H. armigera larvae, by interferring with midgut trypsin-like activity that was found higher in III-IV ages than other ages (Caccia et al. 1999, work presented at the workshop “Resistenza durevole a stress biotici nelle piante: contributo delle biotecnologie”, Tuscia University, Viterbo, 20-21 May 1999; Schettino 2004). The introduction of the CpTI (cowpea trypsin inhibitor) gene into tobacco, showed that its expression in the transgenic plants resulted in a significant reduction of damage by another lepidopteran Noctuidea Heliothis virescens. The mean weight was generally higher when larvae were fed with BG-106 transformed leaves. No statistical difference in mortality was observed between larvae reared with control (RIG) and with Atcys transgenic tomato (BG-106) leaves. The percentage of adults emerged from the cocoon was 81% and 76% for the control and BG-106 respectively. The sex ratio (males/females) was in favour of the female sex both for the RIG (0.94) and BG-106 (0.79) cocoons. On average, the number of laid eggs of the BG-106 fed females was 33% lower than the control. By considering the percentage of hatched eggs (emerged larvae), the value obtained was 6.8% for BG-106 against 11% for RIG. According to these data, in Atcys transgenic tomato a level of cystein proteinase inhibition double than the untransformed control is sufficient to negatively influence the H. armigera biological cycle in particular on number of eggs laid and on fertility (percentage of emerged larvae per female), even if the weight of the larvae fed with the BG-106 leaves is on average higher than the control (RIG). The last datum is in agreement with similar experiments reported in literature where the effect of proteinase inhibitors is tested in different host-pest systems (Ryan 1990). Tomato plants transgenic for genes coding for protein inhibitors, were tested in order to evaluate their capability to inhibit the commercial proteinase activities as well as those present in the midgut of the phytophagous insect H. armigera. Leaf extracts of Pi-IV (gene from soybean coding for a Bowman-Birk-like trypsin inhibitor) transgenic T3 plants (cv. UC-82) were able to inhibit bovine pancreatic trypsin and trypsin-like activity of the phytophagous insect four times more than control plant extracts. Extracts of KTI3 (gene from soybean coding for a


Kunitz-like tripsine inhibitor) transgenic T3 plants (cv. Riogrande) were able to inhibit bovine pancreatic trypsin and trypsin-like activity of the insect 200 times more than control plant extracts; leaf extracts of AtCys (gene from Arabidopsis coding for a cysteine protease inhibitor) transgenic T2 plants (cv. Riogrande) were able to inhibit commercial papain and cysteine-like activity of the insect four times more than control plant extracts (Dirollo 1998). On the basis of the data it would be interesting, for both speculative and applicative purposes, to use these transgenic plants in order to verify in vivo the action on the mortality and/or on the reproduction ability of the H. armigera, particularly dangerous to the tomato crop (Farisei et al. 2002, XIX National Congress of Entomology, Catania 10-15 June 2002). Aim of a further work was to perform a bioassay on the cotton bollworm, feeding the larva with Kunitz proteinase inhibitor coded by the KTI3 gene transgenic plant leaves. The molecules belonging to the Kunitz family have a molecular weight of 21-24 kDa and inhibit the serine proteinase, resulting active in the pH interval 9-11, that is the distinctive interval of Lepidoptera larva midgut. Cv Riogrande tomato plants were transformed with the KTI3 gene through Agrobacterium tumefaciens. The analysis of the data obtained and their elaboration show a direct negative effect on the Lepidoptera biology, particularly in the second generation; this result could have, in the future, a direct containment action of the phytophagous populations in nature, causing a relevant decrease of the marketable production cull percentage (Fig. 4; Speranza et al. 2002 XIX National Congress of Entomology, Catania 10-15 June 2002).

high pigment (hp) see Tomato, “high-pigment genes”

high pigment-2 (hp-2)

see Tomato, “high-pigment genes” Induced variation, physical and chemical

Andrea Mazzucato (Università della Tuscia, Viterbo Italy) Francesco Salamini (Parco Tecnologico Padano, Lodi Italy) Soressi’s research activity begun in the early ‘60s, when he worked on his thesis on the effect of X rays as mutagens for tomato seeds (Soressi 1964). The first publications described this mutagenic approach considering different genotypes and irradiation dosages on traits such as germination, seedling height, time to flowering and ripening, plant height, fruit size and number of seeds per fruit (Bianchi et al. 1963; 1964). The effect of gametic and of intrasomatic (diplontic) selection was described in detail (Bianchi et


al. 1963). The first mutants derived from these experiments were described in detail, such as gamosepalous (fused sepals), typically sterile (aborted inflorescence), skin finely wrinkled (rough fruit epidermis), cabbage leaf-2 (large and blistered leaves), finely netted (leaves paler and netted) (Soressi et al. 1966; Cravedi 1966; Soressi and Cravedi 1967). Soon after, experiments with chemical mutagenesis were started; treatments with ethylmethanesulphonate (EMS) were used on San Marzano and cv Sioux seeds and the general effects on the traits mentioned above described (Marchesi and Soressi 1965). Interesting mutants were described such as variegated (then renamed ghost-2, leaves with regions pale-green of yellowish), turbinate corolla (petal tips turned), lazy-2 (stems prostrated), reticulate virescent-2 (leaves yellowish with green veins), wrinkled leaflets, male sterile-42 (potato leaf male sterile), short anthers (reduced anther segments, see parthenocarpic fruit), hp-2 (high pigment) (Soressi et al. 1966; Cravedi 1966; Soressi and Cravedi 1967; Cravedi and Soressi 1969; Bianchi and Soressi 1969; Soressi 1970a; Soressi 1975). Later, the effects of combining chemical mutagenesis with in vitro induced variation was studies (Mensurati 1994, see Induced variation, somaclonal) In Phaseolus vulgaris, dormant seeds were treated with different doses of EMS and several variants in chlorophyll content and seed morphology and colour obtained. The M1 plants were chimerical, often having only a small mutated sector. Based on the segregation ratios obtained, the number of initial cells present in the apical meristem ranges from three to eight for the whole plant and from one to three for the primary branches. The experimental results indicate a post-treatment origin of the primary branch initials. No evidence was found for the occurrence of diplontic selection during the ontogeny of a mutated plant in the variety studied (Motto et al. 1975).

Induced variation, somaclonal

Francesco Mensurati (Università degli Studi della Tuscia, Viterbo Italy) Antonella Portesi, Cristina Cunico, Elena Perri, Giuseppe Stancanelli (Università Cattolica del Sacro Cuore di Piacenza, Italy) Roberto Ranieri (Stuard experimental farm, Parma Italy) The genetic changes occurring in somatic cells during adventitious meristem formation and shoot regeneration is referred as somaclonal variation. The first approach for setting up the regeneration procedure, with Soressi’s supervision, started in the 80’s at the Piacenza Botany and Plant Genetic Institute (Antonelli et al. 1987; Portesi et al. 1989, XXXIII SIGA Annual Congress, Sassari). Perri (1991) studied the effect of culture medium composition, explant type and the starting genotype on the


frequency of regenerants and induced variations. Cunico (1991) described and characterized the most interesting somaclones in particular the first “bitter fruit” (bf) mutant, singled out in S. lycopersicum. A contribution for innovating nursery in horticulture is reported in Soressi et al. (1991). To have an effective comparison between the segregant progenies of the regenerated plant and its true control plant, the rescue of the cotyledon explant donor seedling was done by in vitro rooting of the root-deprived hypocothyl. Following the cv. Alice cotyledon explants regeneration, at the University of Parma, the percentage of R1 progenies segregating mutants, including seedling lethals, was 11% of the control and 48% of the R2 from regenerated somaclones (Tiburtini 1992). The unusual frequency of twins (from polyembryonic seeds) observed in the R2 progenies, was attributed to an hormonal influence during the sexual embryogenesis in the R1 regenerated plant (Cunico et al. 1992, XXXVI SIGA Annual Congress). A further experiment (1988-1989) in collaboration with the University of Parma and the support of the Stuard farm was accomplished by plant regeneration from hypocothyl cotyledon and leaf callus of cvs. Alice and S. Marzano. The data confirmed somaclonal variation as a powerful tool for creating genetic variability (Ranieri et al. 1996). By excluding polyploids, 26% of Alice progenies and 9% of S. Marzano showed mutations which were consistent through generations. Among the chlorophyll mutants the mottled green fruit shoulder (um) was described for the first time. Certainly, highly informative data on somaclonal variation came from the PhD thesis of F. Mensurati (1994). In this work, by recovering the cotyledon explant donor seedlings, the mutagenic effect of EMS seed treatment and cotyledon regeneration of EMS treated and non treated seeds, was evaluated in a monostem (to-2) line and in the cv. Alice. The procedure adopted permitted to compare, per each genotype, the distinct segregating progenies from the control (R0S1), the EMS treated seeds (M2), the cotyledon regenerated control (R2) and the regenerated cotyledons of seedlings from EMS treated seeds (R2M2). The main results are dealing with the M1 and R1 plant chimaerism, the low (5%) frequency of polyploid phenotypes among the regenerated plants, the mono-Mendelian recessive mutant frequency that resulted significantly lower in the cotyledon explant regeneration than that following EMS seed treatment. The combination of EMS seed treatment with regeneration of cotyledon-derived seedlings from EMS-treated seeds allowed to recover mutants that would otherwise be lost by merely selfing the M1 plants. This combined procedure succeeded in the screening of a cv. Alice anthocyaninless mutant induced by EMS seed treatment, but recovered through its cotyledon regeneration. In fact, in the literature on tomato somaclonal variation via regeneration no


anthocyaninless mutant has been found in opposition to that occurred following EMS seed treatments (see Figure).

In vitro culture in vegetable species

Enrico Santangelo, Riccardo Caccia, Emilio Mendoza-de Gyves (Università della Tuscia, Viterbo Italy) Paula Bima (Facultad de Ciencias Agropecuarias, Universidad de Córdoba, Argentina) In the present Laboratory of Biotechnology of Vegetable Species at DABAC, tissue culture has become a routine approach for breeding

purposes. The contributions here cited played a relevant role in achieving not negligible results. Efforts were made to develop more efficient methodologies by refining culture media for plant micropropagation,

Fig.1 Tomato shoot regeneration fromhypocotyl explants.

Fig.2 Tertiary axillary shootemerged from nodalmicrocutting; arrows:cutting sections of theprimary and secondaryshoots previously formed.

Specimen of recessive mutants recovered in segregating progeniesfollowing cotyledon regeneration, EMS seed treatment and theircombination: sublethal yellow and WT seedlings, a; sublethal twins frompolyembryonic seeds, b; adult plant of the bitter fruit (bf) mutant withyellow calyx trait, c; seedlings without (left) or with (middle and right)anthocyanin, d.


regeneration and transformation of different plant species such as tomato, garlic, bean, chickpea and others. Moreover, in vitro cultures were utilized also in basic research. Most of the micropropagation, cotyledon explant regeneration and transformation procedures applied, also involving physiological analyses, were carried out on tomato. Sources of tolerance to abiotic stresses were screened among shoots of genotypes differing in salt sensitivity regenerated on media with various NaCl concentrations (Habashy et al. 1993, XXXVII SIGA Annual Congress). Plants tolerant to Pseudomonas syringae pv. tomato not showing the typical necrotic leaf spots due to Fenthion spray, were induced by regenerating shoots on media containing this insecticide molecule (Santangelo et al. 1995-ISPAV, Roma, Italy). Chemical mutagenesis through EMS seed treatment coupled with cotyledon explant regeneration and rescue of the apical meristem from the cotyledon-excised seedlings were successfully employed for increasing the range of somaclonal variants obtainable in tomato (Mensurati et al. 1995, XXIX SIGA Annual Congress). GA3 supplied to culture medium was effective in inducing axillary meristems in shootless monostem tomato (to-2) (Bima et al. 1995). To reduce the polyploids frequency among the shoots regenerated in media supplied with hormones, the capacity of hypocotyl explants to regenerate in hormoneless media was assessed, confirming that both regenerating ability and polyploids frequency are highly dependent on the genotype, not omitting that hypocotyl is a mixoploid tissue (Fig. 1; Bima et al. 1993). On the basis of such indications, to develop triploid tomato plants, the tetraploid parents crossed with the diploid ones have been obtained not by colchicine treatment (Allavena et al. 1976) but through hypocotyl explant regeneration (Habashy et al. 2004). Cotyledonary explant regeneration and vegetative propagation via nodal microcuttings were steps routinely adopted in our experiments of genetic transformation (Schettino et al. 1998). In tomato, each in vitro nodal microcutting produces only one axillary shoot; aiming to obtain supernumerary axillary shoots per nodal microcutting, different media, with or without plant growth regulators, were tested. Both IAA and zeatin were able to gradually induce the proliferation of supernumerary axillary shoots when the previously formed were removed. To avoid the occurrence of polyploid variants among the plants derived from supernumerary shoots, auxin (IAA) is to be excluded in all the steps of shoot proliferation. The sole cytokinin (zeatin) when added to the basal medium gave the highest proliferation efficiency (Fig. 2; Cammareri 2003; Soressi et al. 2008). Efforts for in vitro organogenesis and embryogenesis in Phaseolus vulgaris were made by Allavena and Rossetti (1983, 1986). In the 90’ s in many labs


numerous experiments were planned in bean to set up tissue culture and genetic transformation protocols. To investigate their morphogenetic potentiality, apical meristems, from 24 h water-soaked dry seeds, were induced to form adventitious shoots in presence of 1-2 mM of thidiazuron (TDZ). The data obtained confirm a competence of P. coccineus better than P. vulgaris even dependent on the intraspecific variability; the best proliferating genotypes tested resulted to be cv. Venere and cv. Montecarlo respectively (Ranalli et al. 2001). The need for a reliable in vitro vegetative propagation protocol in garlic encouraged P. Bima, tutored by Soressi, to set up an efficient methodology for micropropagating garlic clones of different origin (Bima 1997).

Insect resistance

Stefano Speranza (Università della Tuscia, Viterbo Italy) The interest of GP Soressi towards insect resistance begun in the early ‘90s when experiments aimed at exploiting the introgression of morphological characters from wild species into cultivated potato were carried out (Remotti 1990). Later, this topic was expecially developed in tomato where the genetic transformation with protease inhibitor genes (Kunits, Bowman Birk and cystatine) was tested as a method for conferring insect resistance. A long and fruitful collaboration with Prof C. Pucci and Dr S. Speranza of the Department of Plant Protection at the Tuscia University allow to test the effect of this genetic improvement towards the tolerance against important tomato noxious insects (see Helicoverpa armigera; see Spodoptera littoralis).

Kunitz gene see Helicoverpa armigera

Landraces, bean

Lucia Lioi (Istituto di Genetica Vegetale, CNR, Bari Italy) Bruno Campion (Unità di Ricerca per l’Orticoltura, CRA, Montanaso Lombardo Italy) Andrea Mazzucato (Università della Tuscia, Viterbo Italy) From the hillside to the shores surrounding the Bolsena lake (Viterbo), within the Municipalities of Gradoli and Acquapendente, since long ago has been cultivated the bean landrace “Fagiolo del Purgatorio”, so called because traditionally dined during the Purgatory luncheon of Ash Wednesday. This crop covers an area of ∼ 20 ha, yielding 8-10 q/ha of dry grain. The research activity, carried out in the framework of the MIUR-


MIPAF project “Valorisation of the Italian vegetable germplasm” and of a research contract with the Comunità Montana Alta Tuscia Laziale, aims to characterize both the bean plant and its product together with the phytosanitary healing. Bio-agronomic trials were developed by comparing the seed accessions provided by local farmers. On the initial seed samples and on those reproduced in open field at Viterbo, biochemical (seed storage proteins), nutritional and molecular-genetic (SSR and AFLP markers) analyses were carried out (Santangelo et al. 2006; Lioi et al. 2007). The results revealed the extent of genetic diversity in this bean landrace and permitted to select homogeneous, representative accessions. Moreover they have evidenced the Meso-american gene-pool origin of the “Fagiolo del Purgatorio” and the presence of two plant growth habits (bush determinate, bush indeterminate) highly correlated with the seed storage protein phytohaemagglutinin (PHA) electrophoretic variants SG2 and MG2 respectively. The two distinct growth habits differ in plant height, number of nodes and pods, pod length, seeds per pod, earliness and dry grain yield and in addition in sensitivity to pathogens. In collaboration with the “Gian Pietro Ballatore” Consortium and in the framework of the “Piano per la produzione di proteine vegetali (3P-Sicilia)”, a morphological and molecular analysis has been carried out on the landrace from Sicily “Fagiolo Badda”. Compared with other Italian landraces, this characteristic bi-coloured bean showed a very late flowering time and a ripening of pods in October. Molecular analyses (ISSRs) indicated that the Badda bean belongs to the Andean gene-pool and is distinguishable from other borlotto types or bi-coloured varieties (Paniconi 2006).

Landraces, lentil Andrea Mazzucato, Pietro Mosconi (Università della Tuscia, Viterbo Italy) Research on lentil has been recently carried out in collaboration with two public consortia. The Comunità Montana Alta Tuscia Laziale committed a molecular characterization of the landrace “Lenticchia di Onano” from Northern Latium, whereas the “Gian Pietro Ballatore” Consortium in the framework of the “Piano per la produzione di proteine vegetali (3P-Sicilia)” commited the characterization of three landraces from Sicily (“Lenticchia di Linosa”, “Lenticchia di Ustica”, “Lenticchia di Villalba”). Several accessions were fingerprinted by ISSR markers; all the landraces clustered separately (Gianfilippi 2006). For the “Lenticchia di Onano” landrace, the molecular analyses confirmed the suspicion that actually two different morphological and molecular typologies are cultivated and commercialized


as “Onano” landrace. Only further molecular and historical investigation could reveal which is the true original type.

Landraces, tomato Andrea Mazzucato, Giorgio Chioccia (Università della Tuscia, Viterbo Italy) Research has been carried out on several tomato landraces with particular focus on those cultivated in Latium such as Scatolone di Bolsena, Pantano romanesco, Spagnoletta del golfo di Gaeta e Formia (Mazzucato et al. 2006). Scatolone is a landrace cultivated on the hills surrounding the Bolsena lake with very good organoleptic qualities and a characterisc boxiness (scatolatura) of the fruit. An extended germplasm collection has been done in 1994 and a mild selection has been done among 42 accession from ten different farms (Pepponi 1995). The landrace is characterized by the occurrence of vegetative, “male” individuals that correspond to the “rogue” phenotype described in some British and Dutch cultivars; such character shows a weak genetics basis and the only way to reduce the occurrence of rogues is to keep low the germination temperature (Mazzucato et al. 1998a). A selection of Italian landraces showing high differentiation for fruit shape, size and utilization has recently been used to identify marker-trait relationships in an association mapping approach (Mazzucato et al. 2008a; see Association genetics). Also very recent is a collaboration with the CRA-Unità di Ricerca per l’Orticoltura of Monsampolo del Tronto (AP) for the molecular characterization of the “A pera abruzzese” landrace. The Soressi’s group has been referenced by the Regional Agency for Agricultural Development for the inclusion of tomato landraces from Latium in the “Registro Volontario Regionale delle risorse genetiche autoctone della regione Lazio”.

“Lesion mimic” mutants Enrico Santangelo, Mariagrazia Antonelli (Università della Tuscia, Viterbo Italy) “Lesion-mimic” mutants, mimicking cell death, are a powerful tool for the comprehension of mechanisms activated during plant stress response. Soressi singled out in a progeny under selection of Dr. Svetlana, Moscow, a tomato plant (V20368) that, at increasing temperature and high light intensity, exhibited leaf necrotic lesions, with acropetal progression. The autonecrosis resulted due to the interaction of the Cf-2 resistance gene


(from Solanum pimpinellifolium) with Rcr3esc gene, encoding for a cysteine protease, of S. lycopersicum (Santangelo et al. 2003a). The mutant revealed to be a valuable tool for studying molecular processes common to different stressing agents. The Cf2/Rcr3esc interaction is already

active in asymptomatic leaves of the autonecrotic line V20368, causing in turn a strong oxidative burst (Santangelo et al. 2003a; Santangelo et al. 2007a). A delay, or a lacking, in the activation of the defensive barrier has been observed by Marabottini et al. (2007, LI SIGA Annual Congress): scavenging enzymes of H2O2 (catalase) were functioning, but not all those committed to superoxide removal; the system of

ascorbate-glutathione cycle was at levels of non-stressed controls; markers of PCD and HR were both activated during the autonecrotic process. Chloroplasts and mitochondria appeared directly affected by oxidative stress. Considering the pivotal function of ethylene signalling in response to stresses, Fonzo et al. (2003, XLVI SIGA Annual Congress) investigated its role in modulating the necrosis formation by crossing the V20368 line with the ethylene tomato mutants Nr, rin and nor (see Ethylene). An ethylene burst resulted evident when the autonecrotic process was triggered in V20368, but in the F1 hybrid with the WT plant modifications in some steps of the ethylene metabolism altered and modulated the phenotypic expression of the autonecrosis. A consistent maternal effect appeared, thus supporting the hypothesis of an involvement of mitochondria in the triggering and spreading of the autonecrosis. In V20368, Santangelo et al. (2007b) observed a remarkably higher level of HCN associated with an ethylene burst and supposed an overwhelming of the detoxifying capacity of β-CAS (enzyme of mitochondrial fraction detoxifying cytosolic CN-) and AOX1b (mitochondrial alternative oxidase) that led to an inhibition of mithocondria respiration, ROS overproduction and cell death. When treated with ozone, the mutant line V20368 (but not its control) developed, 96 h after the fumigation, O3-dependent necrotic lesions associated with a significant increase in both foliar ethylene release and CN- levels (Antonelli et al. 2007). Finally, the histochemical analysis of V20368 leaves treated with Fenthion revealed, the presence of dead cells

Sympomatic leaves of the V20368autonecrotic line (Santangelo et al.2003a).


(absent in the control RioGrande) thus confirming its greater sensitivity to abiotic stresses (Antonelli et al. 2006, XIII SIPAV Annual Meeting).

lutescent-2 (l-2) see Tomato, “high-pigment genes”

MADS-box genes

see parthenocarpic fruit; stamenless-2

Maize, hybrids Carlo Lorenzoni (Università Cattolica del Sacro Cuore, Piacenza Italy) Compared to that in tomato or other vegetables, Soressi’s activity in maize genetics and breeding has been limited. In a report dated 1972, comparison data for 725 opaque-2 hybrids are presented. The yield trials were located in the Po Valley at Cremona, Castelnuovo B.A. (Milano), Pozzuolo (Udine). As a result of these experiments, the authors identified some pedigrees that reduce the well known disadvantages of the opaque-2 maize (Maggiore et al. 1972).

Male sterility see 7B-1 mutant; pistillate; positional sterile-2; stamenless-2, parthenocarpic fruit

Microgravity Andrea Mazzucato (Università della Tuscia, Viterbo Italy) In a colaborative research with the group of Prof. F. Saccardo, a horizontal uniaxial clinostat has been used to grow tomato plants under simulated microgravity conditions. The study was carried out to evaluate the vegetative and reproductive behaviour of the dwarf tomato variety ‘Micro-Tom’ in simulated microgravity conditions and to determine if tomato plants would complete their life cycle (‘seed-to-seed’). Plants grown under simulated microgravity exhibited a spreading growth and an increased internode length. Total fruit yield, leaf area, leaf dry weight, fruit dry weight, total dry weight and shoot/root ratio were lower in the clinorotated tomato plants than those grown in the control treatment. The amount of foliar carotenoids, and chlorophyll a and b were also substantially reduced under simulated microgravity conditions. Quality parameters (total soluble solids and fruit dry matter) of tomato plants were also negatively affected by clinorotation. Although the number of flowers was increased, fruit setting was reduced by 46% under clinorotation. However, no significant


difference was recorded for the pollen fertility and the seed number per fruit. Seeds produced by clinorotated plants were germinated proving to be biologically and functionally normal, thus indicating that ‘Micro-Tom’ plants could realize complete ontogenesis, from seed to seed, in microgravity (Colla et al. 2007).

Monostem for tomato processing Bruno Campion, Massimo Schiavi (CRA- Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) Piero Frangi (Fondazione Minoprio, Como Italy) Giuseppe Gandolfi (Università Cattolica del Sacro Cuore di Piacenza, Italy) According to Soressi, the persistent problem of processing tomato mechanical harvesting in Italy during the the ‘60-‘80s could be got over by

putting in practice the monostem ideotype represented by a short, possibly erect plant without lateral shoots, bearing only one truss with 3-5 firm fruits, to be grown at high density population (Soressi 1974, 1975a, 1976). Such an ideal plant should result in a tomato crop with an early “simultaneous” agronomic ripening due to the short flowering period and the

reduced number of fruits per plant counterbalanced by a high number of plants per unit area. By trying to select such a kind of plant, the known mutants lacking or having few lateral shoots were collected and characterized. Some of them, namely blind (bl), lateral suppressor (ls), torosa-2 (to-2), eramosa (era), unicaulis (uni), were crossed with some breeding lines and commercial cultivars (Petomech, CaJ, XP1030). Up-right plants, reduced in size, without lateral shoots, bearing 1 truss with 1-2 seeded fruits, therefore close to the monostem ideotype, were found in the F2, from the crosses involving only bl or to-2. Plants with analogous habit werw found in the F2 progeny segregating ls but because of

Fig.1 High density field plot withmonostem plants at fruit turning stage.

Fig.2 Monostem early, ripe plant ofthe 2nd selection cycle.


their smaller fruits, variable in seed number and size, they were not considered. Following crosses between the best to-2 lines and in few cases backcross to seven commercial cultivars, different monostem (STU) lines were chosen. On the basis of an allelismo test (Schiavi et al. 1980), to-2 and bl mutants resulted to be attributable to same locus and their expression influenced by the background effect. The F2 to-2 plants judged particularly interesting for the programmed breeding purposes were crossed and backcrossed to 13 commercial processing varieties. Among these pre-breeding stocks, the first 17 S4 monostem (STU) lines were selected (Schiavi, 1977) and agronomically evaluated for earliness, concentrated ripening, adaptability to high density population (Fig. 1) in comparison with the cvs. Ventura and UC105 (Campion 1977). Some of them were used for studying growth parameters (LAI, LAR, NAR, HI, SL, DW) in comparison with cv. UC82 (Frangi 1984; Frangi and Soressi 1986) and ones as NILs for investigating physiological and biochemical aspects (Soressi and Mapelli 1992). The same monostem line was used to evaluate its response to various plant population density, ranging from 2.75 to 8.8 plants/sqm (Soressi et al. 1986). The breeding activity went on by selecting the progenies from the inter-crosses of the best monostem lines (Fig. 2) derived by the previous crosses with 13 commercial varieties (Bruzzone et al. 1984). The build-up and distribution of the dry matter and some of its components in both monostem and normal genotypes were also examined (Chiappari 1993). Potential and limits of the monostem type to be exploited for the tomato breeding was analyzed by Scolari (1987) who evaluated earliness and quality of the raw and processed product. In order to increase the number of flowers/fruits per truss, the interaction of to-2 and s (compound inflorescence) genes was examined in segregating progenies, remarking an epistatic effect of to-2, so impairing next efforts. Altogether, three cycles of selection were carried out by combining the best monostem lines from the 2nd cycle. At the end of the 3rd selection cycle some of the best performing STU lines were evaluated during 1988-90. Gandolfi (1992) characterized the most interesting monostem (STU) breeding lines, giving also their pedigree through 4 cycles of subsequent selection. All these breeding lines are earlier (10-15 days) than Red Setter, with equal or superior quality (pH, ° Brix and Bostwick) but inferior for yield. Therefore, none of the breeding lines selected was suitable for wide field cultivation in relation to the high plant density needed (250-300 thousand/ha). For reducing such a high density plant population, an higher fruit number per plant (from 2-3 to 4-5) should be required. However, some indications have emerged about the possibility of using the earliest monostem lines for a 2nd tomato field culture (after pea or barley) without yield reduction. A parallel selection work followed crosses between to-2 and pat-2 (Poma 1992). Poma contributed to select, evaluate and describe a number of monostem


(to-2), parthenocarpic (pat-2) lines of possible interest for both table and early processing tomato.

Monostem in protected cultivation

Pardossi Alberto, Tognoni Franco (University of Pisa, Italy)

The breeding work with monostem breeding lines for open field cultivation prompted Soressi to elaborate a new plant model for energy saving in greenhouse (Soressi and Scarascia Mugnozza 1988) and for growing tomato plants particularly suitable to soilless culture techniques (Soressi et al. 1988). Taking into account the difficulties of pollen dehiscence and fertilization in flowers of

monostem plants in NFT culture (Pardossi et al. 1988), in collaboration between Tuscia University and Pisa University, glasshouse trials with monostem parthenocarpic (pat-2) lines were carried out by adopting the NFT culture system. In fact, in Mediterranean winter-spring condition, the pat-2 gene interacts favourably with to-2 not only improving flower set but also increasing fruit size (Baroni 1994). Positive features, constraints and perspectives are reported in Soressi et al. 1992.

“Never ripe” mutants Enrico Santangelo, Maurizio Enea Picarella (Università della Tuscia, Viterbo Italy) The tomato “Never ripe” group includes the mutants Never ripe (Nr),

ripening inhibitor (rin), non-ripening (nor) and nor2 (Soressi 1975b) with an altered synthesis and/or perception of ethylene, displaying a phenotype with delayed ripening process and a

modified fruit pigment synthesis and production. The dominant mutation Nr encodes for an ethylene receptor, determining the production of fruits

Fruits of wild type (+/+) and ripening mutants at full ripening.

Monostem, non parthenocarpic, poorfertile line (a), correspondingparthenocarpic pat-2 line with improvedfertility and fruit size (b).


that do not fully ripen. rin results from a deletion in the last exon of a tomato MADS-box transcription factor gene designated LeMADS-RIN; the ripening process is blocked before the climateric burst and the fruit has a slow ripening becoming lastly light yellow. The nor locus harbors a gene with structural features suggestive of a transcription factor. The nor2 mutant, allelic to nor, had been collected in 1968 by Soressi in a vegetable garden of a grower who considered it as a “falling-in-love” type for an unexplicable (to him) occurrence of a plant with red fruits among the yellow-orange nor2 “winter type”. Both rin and nor genes behave as incompletely recessive for both fruit colour and biochemical level. Indeed, while positively affecting the fruit shelf-life, these heterozygous in opposition to the homozygous genotypes, do not affect so negatively the fruit colour. In Italy, the first biochemical investigation on fruits of the “never ripe” mutants (Nr, rin and nor2) and high pigment (hp and hp-2) and green flesh (gf) has been promoted by Soressi (Martiniello 1975; Palmieri et al. 1975; Palmieri and Soressi 1978). In these papers the authors stressed the theoretical and practical interest in particular of hp-2 and nor2 to raise the vitaminic content and to naturally extend fruit shelf-life and storage without loss of nutritional value. The influence of Nr and mainly nor2 alleles on quality and storage of fruits from different table and processing cultivars and their F1 hybrids with the same two mutants was studied by Soressi’s ISPORT group and IVTPA, Milan (Palmieri et al. 1976; Rossi 1977; Sozzi et al. 1981). A key contribution in this topic is represented by the two field experiments in F. Rossi’s dissertation (1976) with fruit storage trials at room and controlled (12°C, 80-90% R.H.) conditions. Productive and quality field data revealed the nor2 allele affecting, even when heterozygous in F1 hybrids, the shelf-life of the on-vine tomato fruits, so allowing a greater ripening concentration of commercial yield in once-over harvest. Shelf-life and storage values, colour, lycopene, beta-carotene, chlorophyll, sucrose, reducing sugars and vitamin C contents of 9 F1 hybrids (4 Nr/+ and 5 nor2/+) were reported in Palmieri et al. (1976) and Sozzi et al. (1981). These authors put in evidence notable advantages expected by exploiting the nor2 heterozygote in F1 tomato hybrids suitable to mechanical harvesting and processing with high quality fruits. The effect of the nor2 mutant in Gimar background on the fruit quality during on-vine ripening, over-ripening and post-harvest was investigated in Lancellotti’s graduation thesis, then published in Allavena et al. (1978). The recorded traits were: days from flowering to green mature, turning stage, ripening and four subsequent overripening stages; °Brix, dry matter, acidity, and the same abovementioned traits (total carotenoids, etc.). These authors stated that the better shelf-life, firmness and quality of the Gimar nor2/+ fruits, either on-vine or post-harvest, offers encouraging perspectives for delaying in time and space the consumption of fresh fruits or their processing.


On the basis of such results and perspectives discussed with Soressi, Nira Retig supplied with nor2 seeds, stimulated breeders from Israel to develop F1 tomato hybrids, heterozygous for nor2 or rin, producing firm, long-storing fruits, both on-vine and post-harvest. Naomi (Hazera Seed company) was the first F1 nor2/+ cherry tomato hybrid, rapidly spread and grown from Sicily to the North of Italy. These Israelian nor2/+ and rin/+ F1 hybrids changed the way of harvesting tomato, from picking fruit by fruit at turning stage, into truss by truss with red fruits, thus opening the era of the long shelf-life red-ripe tomatoes.

non ripening (nor) see “Never ripe” mutants

non-ripening3 (nor3) see “Fruit skin” mutants

Onion Massimo Schiavi, Bruno Campion (CRA- Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) At the beginning of the 1970’s the authorities of Parma Province supported the formulation of a breeding program for the onion variety “Dorata di Parma” by the Institute of Botany and Plant Genetics, run by Prof. Lorenzoni, at Piacenza University. The aims were to solve the main problems found in the populations grown locally (high sensitivity to Fusarium basal-rot and lack of bulb uniformity). The Emilia Romagna Region’s Department of Agriculture also recognized the importance of this crop in its territory and agreed a separate program for onion genetic improvement with the Experimental Institute for Vegetable Crops of Montanaso Lombardo (LO) under the Soressi guide. Considering the similarity of the two programmes, both the Institutes recognized the benefits from joining the two forces and requesting the collaboration of the Institute of Plant Pathology, University of Bologna, that had already begun studies on onion phytopathological constraints. Afterwards, the Italian Ministry of Agriculture increased the funds for the onion programme within the “Orticoltura” project (Soressi and Porcelli 1988), sub-project “Asparagus and onion” (Soressi 1988a; 1988b). The selection methods adopted were: mass selection on naturally infected soil and three systems of recurrent selection with evaluation of S1, S2 and half-sib progenies both on infected soil and by means of artificial infection (Schiavi et al. 1984d; 1984e; 1988a). Within these activities a different pronubous efficacy of honey bee towards blowfly in controlled onion pollination was ascertained (Fig. 1; Schiavi et al. 1988b). After three cycles


of selection the improved populations showed a percentage of rotten bulbs twice lower than that of the commercial populations of Dorata di Parma and similar to tolerant commercial hybrids (Fantino and Schiavi, 1987). Progress was also observed in bulb characteristics: uniformity of shape, colour, shelf life and scale covering. In addition, some researches were carried out to study methods for onion androgenesis (Campion et al. 1985, XXVIII SIGA Annual Congress), here reminding

Campion’s later success with onion gynogenesis. At the end of the breeding program, five new cultivars were released: “ERSO 1”, “ERSO 2”, “Marica” and “Morgana” (Fig. 2) of the type “Dorata di Parma” and “Sorriso” derived from the “Borettana” variety (Schiavi et al. 1991). Two graduation theses were produced on evaluation of different techniques of onion seed production including “seed to seed” procedure: Moretti (1982) and Donati (1980).

Ozone Mariagrazia Antonelli, Enrico Santangelo (Università della Tuscia, Viterbo Italy) Maurizio Badiani (Università Mediterranea di Reggio Calabria Italy) Ozone (O3) is the most widely spread atmospheric pollutant in the

industrialized world. When O3 concentration exceeds the tolerance threshold for a given plant, it results in the formation of visible damage, which has been described as necrotic lesions, bifacial flecking, bronzing, and "waterlogging". At full development, these symptoms physically resemble HR lesions. O3 requires entry

One hundred nL L-1 ozone for 5 h causesvisible symptoms in the lesion-mimicmutant V20368 (left) but not in itsphenotypic control line MP-22 (right).

Fig.2 Morgana: an improved onionvariety tolerant to Fusarium basal-rot.

Fig.1 Caged blowflies for onion selfing.


into the apoplast through the stomata to exert its toxicity. Once inside the leaf, O3 reacts with the cell wall, the plasma membrane and apoplastic fluids, to produce a burst of reactive oxygen species, such as superoxide radicals, hydrogen peroxide, and hydroxyl radical. Therefore, O3 behaves as an abiotic elicitor in plants, thus suggesting the possibility of using such photochemical oxidant as an experimental tool for studying plant defence responses towards biotic and abiotic stresses. A fruitful collaboration among the universities of Reggio Calabria, Pisa and Perugia, supported by MIUR from 2003 to 2007, elucidated some molecular events and the complex cross-talk among hormonal signalling routes, deployed in tomato upon O3 exposure. Tomato mutant lines impaired in ethylene perception Nr (Never ripe) or biosynthesis rin (ripening inhibitor) (see Ethylene), the tomato autonecrotic line V20368 (see “Lesion mimic” mutants) and anthocyanin mutants (see Anthocyanins) were used as model systems. In a first contribution, it was showed that both the Nr and rin mutations affect activation timing and relative transcripts abundance of the ethylene biosynthetic enzymes 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase, as well as of the ethylene receptor NR, in response to realistically elevated O3. Since both mutants and their near isogenic WTs were equally damaged by the O3 treatment, a sort of isoform "compensation" among ethylene biosynthetic genes could be supposed, with the common aim of producing an ethylene burst triggering defence responses. Such a burst was indeed observed, and showed similar time patterns in the three genotypes, although the peak of ethylene emission levels were different (Antonelli et al. in press). Also the autonecrotic line V20368 revealed to be a valuable tool for studying common molecular responses to different stressing agents in tomato. In response to O3 fumigation, V20368 plants showed a high level of ethylene release and increasing CN- level compared with the control line. Such results support once more the link between an O3-dependent ET burst and the development of foliar symptoms, and suggest that CN- overproduction might take part in the launch of O3–induced PCD (Antonelli et al. 2006). The anthocyanin atv mutant (see Anthocyanins) seems to be more resistant to O3 than other tomato genotypes in terms of foliar symptoms, confirming a role of anthocyanins as ROS scavengers.

‘Panzanella’ see “Fruit skin” mutants


parthenocarpic fruit (pat) Andrea Mazzucato, Luigi Selleri (Università della Tuscia, Viterbo Italy) Dario Beraldi (University of Edinburgh, Edinburgh UK) The parthenocarpic fruit (pat) mutant is the first source of parthenocarpy (fruit set in absence of pollination) described in tomato and was obtained by

Bianchi and Soressi (1969) with mutagenesis with ethyl methanesulfonate (EMS). The mutant phenotype, characterised by a high tendency to parthenocarpy and abnormalities in anther development, was firstly ascribed to the action of two tightly linked genes, sha (short anthers) and pat (parthenocarpic fruit) (Soressi and Salamini 1975). Data demonstrating the practical interest of the sha pat trait for selecting parthenocarpic varieties to be cultivated in open field for processing are reported by De Logu (1974). The discovery of a spontaneous sha-pat mutation

in the line ‘Montfavet 191' (Pecaut and Philouze 1978) proved that the described phenotype was caused by a single recessive mutation with pleiotropic effects and the gene was finally named pat (Soressi 1985; Philouze and Pecaut 1986). The mutant shows an enhanced ovary growth rate during the first 10 days after anthesis, which correlates with a precocious onset of cell divisions in the pericarp and a higher auxin, gibberellin and DNA content in ovaries (Mapelli et al. 1978). At maturity,parthenocarpic pat fruits are typically about two-thirds of the normal size (Falavigna et al. 1978), probably due to decreased cell enlargement, rather than to a lower number of cell layers (Mapelli et al. 1978). Compared to the near isogenic WT, the mutant shows a general better earliness and higher fruit set (Falavigna and Soressi 1987), and a higher content in total soluble solids (Falavigna et al. 1978), therefore raising practical agronomic interest. In addition to parthenocarpy, the pat gene causes aberrations that affect male floral organs, i.e. short, irregular and apparently unfused anthers that leave the stigma exerted (Bianchi and Soressi 1969; Philouze and Pecaut

Fig.1 A fruitful triploid parthenocarpic pat plant.

Fig.2 Variety Partheno plant, showingthe exuberant amout of ripe cherry-typefruits, mostly seedless.


1986). Moreover, while other sources of genetic parthenocarpy are usually facultative (they also can produce normal seeded fruits; Philouze, 1985), female fertility is greatly reduced in pat and seed production is more difficult. More recent research showed that the female sterility of the pat mutant is due to the aberrant development that affects a fraction of the ovules (Mazzucato et al. 1998b) and to overall defects in pollen guidance that occur in the ovary (Mazzucato et al. 2003). Physiological aspects of the mutant phenotype have addressed the role of plant growth regulators, such as polyamines (Antognoni et al. 2002; see Polyamines) and gibberellins (Mapelli et al. 1979; Mazzucato et al. 1999); studies on gibberellin regulation have been extended to a more general perspective on fruit set (Olimpieri 2006; Olimpieri et al. 2007; see Gibberellins). In addition, the pat system has served for studying the interaction with the lateral suppressor mutation (Ziliotti 1984), for the identification of transcripts differentially expressed in parthenocarpic ovaries (testa 1998; Testa et al. 2002; see DDRT-PCR), to synthesize polyploid parthenocarpic lines (Fig.1) (Allavena and Soressi 1978; Habashy et al. 2004; see Polyploidy) and to study the relationship between parthenocarpy and the activity of class B MADS-box genes (Mazzucato et al. 2008b, see stamenless-2). Parallel breeding work led to the constitution and inscription in 1982 of the new variety “Partheno”, a cherry type homozygous for the pat gene, in the National register of varieties (Fig.2). pat is also the first mutation for parthenocarpy in tomato that has been mapped; it is located on the distal end of the long arm of tomato chromosome 3 (Mazzucato et al. 1998c; Beraldi 2002; Beraldi et al. 2004). The ongoing positional cloning project has presently reduced the genomic window containing the Pat locus to a couple of BAC clones, spanning about 170 kb (Selleri et al. 2007, contribution to the LI Annual SIGA Congress). While the list of loci of the ‘pat series’ has nowadays reached the description of pat-9 (Gourguet et al. 2008), the founder member of this family still deserves much attention and promises interesting results to researchers.


parthenocarpic fruit-2 (pat-2) Enrico Santangelo (Università della Tuscia, Viterbo Italy) The fruit setting in tomato early production under cold plastic-tunnel is strongly hindered in Mediterranean areas by low-night and wide fluctuating temperatures, high light intensity and growing daylenght, environmental conditions occurring during spring season. The introduction of

parhenocarpy genes into cultivars of value may encompass such constraints and avoid the use of chemicals. One of the most important source of parthenocarpy is the parthenocarpic fruit (pat-2) gene found in the Russian cultivar Severianin and frequently used in breeding programmes (Santangelo and Soressi, 1990). The interest for such a gene is due to its simple heredity and its capacity to form seedless fruits of comparable size respect the normal ones when the environmental conditions could hamper it, allowing a high stability

and uniformity of production. This mutation has been deeply studied in France by the group of J. Philouze and in Spain by the group of F. Nuez. In Italy, experiments were planned to study pat-2 expression and its effects on productivity in conditions of temperature fluctuating during growth. Different French hybrids were compared in their normal and parthenocarpic pat-2 near isogenic versions (Santangelo 1988; Cantoni 1991). The pat-2 gene allowed an improvement of the fruit setting, when relatively low-night temperature (1st truss) or high-day temperature (4th truss) occurred. Therefore, more stable production and greater commercial fruit size uniformity were obtained. These advantages were more or less pronounced considering both microclimatic conditions or the genetic background where the pat-2 gene is introduced (Bricchi 1993; Mapelli et al. 1995). Because the average number of seeds per fruit never achieves that of the correspondent normal version, such a traits represents a good parameter for selecting parthenocarpic plants (Santangelo 1988; Santangelo et al. 1990). An interaction between the hormonal systems governed by the two different genic situations (WT pat-2) has been observed for the average number of seeds per fruit and the mean fruit weight and diameter. When the environmental conditions are favourable to pat-2 expression, parthenocarpic seedless fruits of optimal size are produced. When the

Mature fruits of a tomato hybrid inhis normal (+/+) and parthenocarpic(pat-2/pat-2) versions.


conditions are more advantageous for seed production, also the parthenocarpic genotypes produce seed, but in a lower number, leading to a light decreasing of fruit diameter. Efforts for mapping the Pat-2 locus have been undertaken by using segregating populations in purely cultivated genetic background (Marras 2002), after crosses with introgression lines (Olimpieri 2006) and with S. pennellii plants (unpublished results).

Pathogenic microorganism culture filtrates for the induction of aspecific plant resistance

Francesco Mensurati (Università degli Studi della Tuscia, Viterbo Italy) Paola Crinò (ENEA, C.R. Casaccia, Roma Italy) Tomato crop cultivations are still requiring chemicals applications to control Phytophtora infestans, causing heavy loss of production, being partial the genetic resistance available for this pathogen. In vitro selection with toxins or culture filtrates of plant pathogenic microorganisms has allowed, in some cases, to improve the efficiency of plant breeding programmes by contributing to isolate disease resistant plants. The use of toxins should be preferred to that of culture filtrates because they are pure substances which do not cause artefacts and do not falsify the results of in vitro selection (Crinò et al. 1996). Under the Soressi’s guidance, and tutored by P. Crinò, Mensurati (1991) attempted to increase through in vitro plant regeneration in presence of the pathogen culture filtrate, the resistance level of tomato cultivars possessing the Ph-2 gene which confers partial resistance to P. infestans. The inoculation essays on seedlings or foliar discs did not evidence any kind of resistance due to the Ph2 gene in comparison to the susceptible genotype, lacking of such a gene. For the experiments planned with culture filtrate or toxin for improving the plant response to P. infestans, the cv. Piline was chosen for its relatively higher number of shoots per explant regenerated in vitro. Therefore, 1290 cotiledonary explants of Piline were cultured on selective medium containing the toxic metabolites of P. infestans. After two subsequent transferrings on substrate containing fresh culture filtrate, 10 non-susceptible plantlets of cv. Piline were in vitro regenerated, rooted and propagated. The work proved a relatively fair efficiency of the in vitro selection procedure in presence of toxic compounds as a simple and rapid way for the identification of resistant or tolerant plants.

peach (p)


see “Fruit skin” mutants Phaselieu Concerted Action

Enrico Santangelo (Università della Tuscia, Viterbo Italy) Ali Akbar Habashy (Agricultural Biotechnology Research Institute, Karaj Iran) The overall aim of the Concerted Action PHASELIEU was to coordinate the on-going research on Phaseolus and to elaborate an integrated strategy model for the improvement of Phaseolus production in Europe for human consumption. The 13 partners participating to the project were representative of Universities and Research Centers of Spain, France, Germany, Belgium, Austria, United Kingdom, Portugal, Israel and Italy. Prof Soressi was the responsible of the Italian group and spokesperson for Italian “Phaseolus workers”: B. Campion (Istituto Sperimentale per l’Orticoltura, Montanaso Lombardo Lodi), D. Bollini (CNR Biosynthesis Lab, Milano) and P. Ranalli (Istituto Sperimentale per le Colture Industriali, Bologna). Collaborative links with R. Papa (Università Politecnica delle Marche, Ancona) and within the framework of the European Cooperative Programme for Crop Genetic Resources Networks (coordinated by L. Maggioni of Bioversity) were established. Within this collaboration, 403 accessions of the Phaseolus collection originally maintained at the University of Cambridge were evaluated and regenerated, by coordinating Italian institutions engaged in bean breeding or research. Inside the Phaselieu network, the Soressi’s team focused its efforts on setting up in vitro protocols for multiple shoots induction and genetic transformation via Agrobacterium or particle gun (see Bean, biotechnological approach). The morphogenic capacity of apical meristems excised from dry seeds was evaluated by inducing them to form adventitious shoots on media containing several combination of TDZ and 2,4-D (in P. coccineus, cv. Venere) or BAP (in P. vulgaris, cv. Montecarlo). The results confirmed a higher regeneration competence of P. coccineus than P. vulgaris. During 5-6 subcultures, 10-15 shoots per explant were obtained for cv. Venere, whereas the cv. Montecarlo did not exceed 4 shoots. The possible use of thin layers of cotyledonary nodes of P. coccineus (cv. Venere) was also investigated. The results showed a good capacity of proliferation allowing to obtain 8-9 shoots per explant after 4 subcultures and indicated this procedure as particularly suitable for genetic transformation by Agrobacterium tumefaciens (Parsons 2000). Provided the induction of multiple shoots from apical meristems in both P. vulgaris and P. coccineus, the group investigated the potentialities of the biolistic system for bean genetic transformation. In 11 independent experiments, 1178 explants were bombarded, whose 193 (16.4%) rooted in selective medium


containing kanamicin and 15 (7.7%) resulted positive to PCR (Santangelo et al. 2000; V Phaselieu Workshop, Hannover). Soressi’s group contributed to the five workshops held during the Concerted Action and to the Final Dissemination Meeting. The Third Workshop on “Biotic and abiotic stresses in bean” was held at Viterbo on 4-5 June 1999.

pistillate (pi)

Andrea Mazzucato, Irene Olimpieri (Università della Tuscia, Viterbo Italy) In this research, we characterized the pistillate (pi) mutant, a genotype that directly recalls mutations affecting class B MADS-box genes, but that has not been further studied after the first description in the middle of the last century. Plants homozygous for the pi allele appear with Mendelian proportions in segregating populations and, compared to wild-type, show delayed flowering, a frequently modified sympodial segment, higher occurrence of compound inflorescences, and reversion of floral meristem to

vegetative identity. The most striking aberration in pi mutant flowers is the homeotic transformation of stamens into carpels. More rarely, homeotic conversions are reported also in the second floral whorl, with staminoid and carpelloid petals. Ultrastructural analysis reveals more or less subtle sepaloid features in the three inner floral whorls, mainly based on the presence, distribution and amount of glandular and non glandular trichomes. In the ovary, a ‘flower within flower’

phenotype was seldom observed; in one instance such phenotype was coupled with the setting of a parthenocarpic fruit, that reiterated the differentiation of a new flower (see Figure; Olimpieri and Mazzucato, submitted). Mapping information positioned Pi on the distal end of the long arm of chromosome 3. According to published maps, this position is not compatible with any of the known class B and E MADS-box tomato genes (Mazzucato et al. 2008b; http://www.sgn.cornell.edu); thus the identity of

Multiple shoots induction in apicalmeristems of cv SL770 (left) andVenere (right) (Parsons 2000).

Parthenocarpic fruit set on a piplant showing a secondaryectopic fruit and sepal or shoot-like outgrowths.


Pi has to be searched among those genes that act upstream to the ABC(DE) system of determination of the identity of floral organs.

Poliploidy Andrea Mazzucato, Enrico Santangelo, Maurizio Enea Picarella (Università della Tuscia, Viterbo, Italy) Andrea Allavena (Res Unit for Floriculture and Ornamental Species, Sanremo Italy) Ali Akbar Habashy (Agricultural Biotechnology Research Institute, Karaj Iran) Polyploid crops are a valuable resource for horticulturists owing to their peculiar advantages with respect to the diploid ones. In tomato, the exploitation of either natural or artificially induced polyploidy turned out to be scarcely promising for the high sterility of triploids and the low yield and high variability in size of the autotetraploid fruits, extremely variable in seed number which is positively correlated to fruit size in this species. Soressi’s research on tomato polyploidy began in the 70’s when he first obtained data evidencing the potential of polyploidy coupled with parthenocarpy for tomato breeding. As reported elsewhere (see parthenocarpic fruit and parthenocarpic fruit-2), parthenocarpy represents

a useful trait for genetic improvement since it allows good yields in environments unfavourable for pollination and fertilization. Soressi and co-workers (Allavena 1975; Allavena et al. 1976; Allavena and Soressi 1978) obtained tetraploid seeds from hand-pollination of polyploid shoots following colchicine treatment of WT and pat/pat F3 seedlings. These seeds produced 4x

plants either parthenocarpic, as fruitful as the corresponding diploid pat/pat, or WT almost completely sterile in greenhouse. The 4x parthenocarpic fruits were larger and heavier (30%) than their corresponding 2x pat/pat. In addition, their soluble solids content and pH did not significantly differ from the pat/pat diploids’, while the ascorbic acid content was higher (20%). Since hormonal treatments of spontaneous tomato triploids gave high fruit yield and quality (Kagan-Zur et al. 1991), a programme to develop genetically parthenocarpic triploid tomato plants through hand-crossing was planned. To this aim Soressi’s team regenerated in vitro, from hypocotyl explants, diploid and tetraploid somaclones belonging to lines carrying pat or pat-2 mutantions and to the respective WTs. The yield recorded in open field plots proved the parthenocarpic polyploid (4x) fruits to be larger than

Representative fruits ofdiploid (left), triploid (centre)and tetraploid parthenocarpicplants (Habashy et al. 2004).


the parthenocarpic diploids (2x), despite their almost complete seedlessness (Kiferle 1998; Habashy et al. 2000, V Giornate Scientifiche SOI). These new 4x and 2x somaclones, in near-isogenic background except than for the pat mutation, were hand crossed each other to obtain triploid (3x) plants. In greenhouse, 3x pat/pat/pat plants yielded seedless and high quality fruits differently from the non-parthenocarpic triploids, completely unfruitful and sterile (Santangelo et al. 2003b; Habashy et al. 2004). This unique, successful result offers a valuable contribution to practically exploit polyploidy in tomato by combining it with genetic parthenocarpy. To tomato breeders, it offers the challenge of the synthesis of fruitful triploid F1 commercial hybrids, with high-quality, seedless, uniformly sized fruits diversified between fresh consumption and processing. Such triploid hybrids, by considering their complete apireny, shall necessarily be vegetatively propagated. Furthermore, the triploid tomato genotypes, even transgenic, unable to produce fertile pollen and seeds, will nullify the risk of gene flow to conventional tomato cultivars (Santangelo et al. 2005, 2nd Solanaceae Genome Workshop, Ischia, Italy).

Polyamines

Andrea Mazzucato (Università della Tuscia, Viterbo Italy) Polyamine (PA) pattern was investigated in tomato floral organs plants at four developmental stages from floral bud until 2 days after anthesis. Differences in putrescine, spermidine and spermine content, both free and conjugated, were evaluated in a wild-type line (WT) and in the parthenocarpic fruit (pat) mutant. PA content was quite similar in WT and pat mutant ovaries, but the two lines showed a different PA pattern throughout flower development. A net accumulation of PAs was observed in concomitance with pre-anthesis floral stages in pat mutant ovaries, while in WT plants the PA pattern did not change significantly during the developmental stages considered. These differences seem strictly related to the physiological events that initiate ovary development, which have been shown to be precocious in the mutant ovaries and occurring at pre-anthesis stages (Antognoni et al. 2002). A decrease in free PA levels and an increase in the conjugated pool were associated with the initiation of cell expansion. The pattern of ornithine decarboxylase, arginine decarboxylase and S-adenosylmethionine decarboxylase gene expression, as well as the respective enzyme activities, were also analysed during development. The lack of correlation between transcript levels and enzyme activity suggests that post-transcriptional control mechanisms must be acting in the regulation of biosynthetic genes (Antognoni et al. 2002).


Pollen-pistil interaction

see parthenocarpic fruit positional sterile-2 (ps-2)

Fabrizio Ruiu (Università della Tuscia, Viterbo Italy) For the production of F1 hybrids, the hybridization of tomato plants is done by manual emasculation and pollination. This is a labour-intensive process and does not guarantee 100% hybridity. There are many genic male sterility systems available in tomato (Badino and Soressi 1976); however they are generally not used on commercial scale because of difficulties with the incorporation into seed parent lines and their maintenance. The recessive mutation positional sterile-2 (ps-2) has been studied from this perspective (Ruiu 2006). The flowers of male sterile plants do not differ from those of

isogenic male fertile line except for the dehiscence phenotype; in the mutant, the stomium does not open properly and the pollen, although viable, is not released (see Figure). The mutant is female fertile.

The expressivity of male sterility can vary with the genetic background and with the environmental conditions, but shaking of the inflorescences does not increase seed yield (Ruiu 2006). This mutation offers the possibility to maintain the female line through manual-selfing, enabling to obtain a complete male sterile progeny. Furthermore, comparative studies conducted in Bulgaria, where the mutation has been described for the first time, on the time necessary for the emasculation of floral buds (as practiced when using fertile seed parent) and flowers at anthesis (as practiced when using a ps-2 line as seed parent), made it clear that emasculation at anthesis was easier and almost two times quicker than emasculation applied on floral buds. The Ps-2 locus has recently been finely mapped by Gourguet et al. (2006).

potato leaf (c) see Tomato, genetic markers

Dehiscent anthers in the WT line WT (left) andfunctional sterile phenotype in ps-2 (right).


Programmed cell death Riccardo Caccia, Andrea Mazzucato (Università della Tuscia, Viterbo Italy) We investigated the senescence of flag leaves of durum wheat (Triticum durum) during grain-filling, and of petal-like ray flowers of Jerusalem artichoke (Helianthus tuberosus) at anthesis. In both systems, we observed cleavage of DNA to high molecular weight fragments, followed by further degradation to nucleosomal fragments (laddering), a classical hallmark of apoptosis. We show that DNA fragmentation in such specialised leaves is

triggered early in organ development, before the appearance of visual symptoms of senescence. Our observations support the hypothesis that senescence and programmed cell death (PCD) are part of the plant developmental program, activated by developmental cues (Caccia et al. 2001). Further research addressed the role of PCD in reproductive tissues and organs; TUNEL analysis on such tissues revealed a common

occurrence of PCD in the anther tapetum, in the nucellus and in the transmitting tissue of the style after pollination (see Figure; unpublished results).

Pto

see Glucose oxidase, GOX; Fen gene

Reporter genes Marianna Rossi, Andrea Mazzucato (Università della Tuscia, Viterbo Italy) The characterization of mutants, whose phenotype is putatively determined by altered hormone levels or perception may shed light on the role played by hormones in different aspects of plant developmental biology, physiology, biochemistry and molecular biology. In this research, the auxin accumulation dynamic has been analysed in a number of tomato floral mutations by means of a reporter construct where the auxin-inducible promoter of the Agrobacterium tumefaciens gene 5 was cloned upstream of the uidA gene (GUS) (Rossi 2002; Mazzucato et al. 2006). The mutants selected for the analyses were: uniflora, lateral suppressor, stamenless 2, fasciated, parthenocarpic fruit-2. All the mutants where crossed with plants

Programmed cell death occurring inthe style transmitting tissue afterpollination (left), compared with anunpollinated style (middle) and aDNAse-treated positive control(right). Black spots represent nucleiwith PCD-controlled DNA cleavage.


harbouring the reporter construct in homozygosis and F2 families were screened for the presence of the transgene (km resistance). From WT and mutant plants segregating in the F2 families, plants organs were collected and subjected to the histochemical GUS assay. Non transgenic specimens, used as negative control, were always devoid of reporter signal. In turn, remarkable, organ-specific differences were reported between mutant and WT plant tissues. WT roots showed strong auxin accumulation at the root tip and in correspondence of the adventitious root primordia. The uf, sl-2 and f mutant showed no difference from the WT at the root level. Differently, pat-2 and ls roots showed a lower IAA content. Signal intensity was also overall associated to root vigour. In leaf tissues, for the WT and all mutants the

signal was absent or very weak, with the exception of sl-2 that showed higher IAA content. The WT ovary showed an increasing auxin content through its development into

fruit, up to a maximum that was reached at the mature green stage. While the uf ovary did not show any difference from the WT, pat-2 and f ovaries and their young fruits had a stronger reporter signal, in agreement with the putative role of the hormone in parthenocarpic development and in carpel number specification. However, auxin content in the pat-2 ovary was lower than in the WT at later stages of fruit development, as it was in the partially sterile ls mutants; such observation strengthens the knowledge that fruit IAA is essentially contributed by the seeds. The reported results on floral mutants and on other developmental tomato mutants have been the object of a PhD thesis (Rossi, 2003). The same reporter construct has been used to study the IAA-GA relationship in plants silenced for the GA biosynthesis gene GA20ox-1 (Olimpieri and Mazzucato unpublished).

ripening inhibitor (rin) see “Never ripe” mutants

IAA accumulation as detected by the p5::GUS reporter construct in the tomato root apex (left), the secondary root emergence zone (middle) and the leaflet (right). The upper row is the negative control.


Salt tolerance Stefania Astolfi, Maurizio Enea Picarella, Ali Akbar Habashy, Enrico Santangelo, Sabrina Zuchi, (Università della Tuscia, Viterbo Italy) Alberto Pardossi (Università di Pisa, Italy) In the frame of Prof. Soressi’s work on plant response to biotic and abiotic stresses, some experiments were dedicated to the salinity stress in tomato. This crop, widely cultivated in the Mediterranean Region requires frequent water supply during spring-summer and this may represent a drawback for its cultivation especially in coastal areas (primary salinity) and greenhouse

(secondary salinity). To succeed in breeding for a complex trait like salt tolerance, the availability of as much reliable as

easy-to-score parameters is crucial. In vitro cotyledon regeneration was coupled with salt stress to induce genetic variability to be screened for sources of tolerance to this abiotic constraint. Nuclear

and cytoplasmic genome alterations were induced in seedlings and plants of R2 generation. Attention was also paid to in vitro vegetative growth parameters such as fresh and dry biomass of root and shoot of seedlings grown on media with different NaCl concentrations (0, 50, 100, 150, 200 mM). Among the genotypes tested, cvs. Cal J and Edkawi proved to be the most tolerant while cv. UC82 the most sensitive to salinity (Habashy 1992; Habashy et al. 1993, XXXVII SIGA Annual Congress, Perugia, Italy). The response to NaCl of these genotypes was confirmed in a further trial in which cvs. Edkawi and Cal J performed better, respectively for root and shoot biomass, than cvs. UC82 and Petomech. In parallel, a positive correlation between such growth response and the expression of specific salt-responsive genes (see Figure), was put in evidence (Picarella et al. 1995, ESF Network-Workshop, Maratea, Italy). Analogous results were reached by an indipendent experiment on the same genotypes but considering early growth rate of 21-days-old plants, grown in greenhouse

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Edkawi UC82 Petomech CalJ

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TSW12 gene

18S-rRNA-normalized, hybridization signal values,obtained through northern blot analysis, of TAS14and TSW12 gene probes, on mRNAs derived fromseedlings grown in 150 mM NaCl. Values arerelative to those from mRNAs of non-treatedseedlings.


with soilless system culture, as a tolerance-related parameter (Pardossi et al., unpublished data). Salt tolerance was also investigated in terms of physiological traits. In particular, ethylene (ET), regarded as a key hormone regulating gene expression of environmentally challenged plants, released from 4-week-old plants grown in a versatile controlled hydroponic system, was measured along with parameters connected with plant salt-stress response like root/shoot biomass, chlorophyll content and others related to the plant nitrogen metabolism, like inorganic osmolites (Cl-, NO3

-), free aminoacids contents and nitrate reductase (NR) activity (Astolfi et al. 2004, XXII SICA Annual Congress, Perugia, Italy). Edkawi, as a salt tolerant cultivar, and cv. Gimar (WT) and its two near-isogenic lines (NILs) carrying the nor2 (Gimar NOR) and Nr (Gimar Nr) mutations, respectively involved in the synthesis and perception of ET (see “Never ripe” mutants) were compared in control (10 mM NaCl) and salinity (70 mM NaCl) conditions. An interesting result was the antagonistic relationship between NO3

- and Cl-, suggesting leaf nitrate content as a marker of plant NaCl tolerance. In fact, the salt treatment caused a considerable increase of leaf NO3

- content only in the tolerant cv. Edkawi, but not in the Gimar NILs. Salinity, then, caused a significant inhibition of NR activity in leaves of Gimar Nr, Gimar NOR and Edkawi plants. In particular, the NR activity increased (+23%) in Gimar WT and was reduced (-75%) in Edkawi, where the highest NO3

- accumulation was measured. Therefore, a very small amount of NO3

- appears sufficient for NR induction while this activity is not induced or even inhibited when NO3

- is beyond a certain threshold. Leaf chlorophyll decline is generally exerted by salt exposure of plants, probably correlated to an indirect effect of NaCl on the content of essential nutrients. In salt-treated plants, whilst a significant decrease in leaf chlorophyll was recorded for cv. Edkawi, Gimar WT and Gimar Nr NILs, a 33% increase was seen in Gimar NOR (Astolfi et al. 2005, XXIII SICA Annual Congress, Turin Italy; Astolfi et al. 2005). Unpublished data proved that the nor2 mutation does affect not only the fruit ripening by suppressing the respiratory climateric, but also the vegetative organs, by reducing ET release in leaf and root of seedlings (4th leaf stage). Moreover, in salinity conditions, the nor2 mutation has a beneficial influence on the content of antioxidants by increasing ascorbate and tocopherol in Gimar NOR fruits, thus improving their nutritional value (Sgherri et al. 2007). As recently advanced by some authors, an increase of the abscisic acid (ABA), another stress-responsive hormone, is required to prevent excess ET production from plant tissue under drought stress. In such conditions ABA synthesis would allow the plant to maintain shoot and root growth. A negative correlation between these two hormones was observed only in the


root system of salt stressed, hydroponically grown, plants (Picarella et al. 2007).

Scatolone di Bolsena

see Landrace, tomato Senescence

see Programmed cell death Spodoptera littoralis

Stefano Speranza, Gianluca Petti, Claudio Pucci (Tuscia University, Viterbo Italy) Considering the biological relationship between anthocyanin (see Anthocyanins) induction and insect attack and the availability of tomato lines differing for anthocyanin content in the vegetative tissues of the plant, experiments have been carried out in order to verify the effect of tomato leaves with different anthocyanin content in the diet of Spodoptera littoralis larvae, from the 4th living day to their pupating. To this aim, it has been used a tomato line with high anthocyanin content in leaves and stems (line V711088, somaclonal variant atvsc), a near isogenic line without anthocyanins (line V711232, somaclonal anthocyaninless variant) and a

normal control line (V710040, cv UC-82). The data obtained during the trial in temperature-controlled environment (see Figure) showed no differences during the exponential growth of the three cohorts. A first difference is seen when the weight peak of the larvae fed with high and zero anthocyanin diet is reached two days in advance than the control.

Differences detected among the three cohorts for larvae survival were not significant. These results suggest that the insect detects, at midgut level, changes in the chemical composition of the leaves, both as an increase of anthocyanins or as an absence, but that such condition is not sufficient to induce death. However, the high anthocyanin content, or anthocyanin absence, although not letal for Spodoptera larvae, could give sub-letal effects to their physiology.

spongy seed (ss)

see Tomato, genetic markers

Weels with tomato leaves feedingSpodoptera larvae.


stamenless-2 (sl-2) Andrea Mazzucato (Università della Tuscia, Viterbo Italy) We addressed the study of the stamenless-2 mutant both as a source of male sterility for hybrid seed production (Badino 1976; Badino and Soressi 1976) and to test the relationship with the pat mutation (see parthenocarpic

fruit) because aberration of the anther cone has been associated with parthenocarpy in tomato and in other species. The two loci were not allelic and the pat sl-2 double mutant showed mainly epistatic effects; sl-2 was epistatic to pat for phenotypes related to male reproductive development (anther phenotype and seed production under open pollination), whereas the pat mutation was the limiting factor for aspects related to the female side (ovule phenotype, fruit size and seed production under hand pollination). Thus, the genetic basis for the floral defects in the two inner floral whorls

is different in the two mutants (Mazzucato et al. 2008b). Genetic analysis indicated that the primary aberration of the sl-2 genotype is the specification of stamen identity, supporting the hypothesis that the Sl locus may correspond to the tomato ortholog of DEF. Although our work supported this hypotesis with mapping information (both Sl and SlDEF mapped on the long arm of chromosome 4, Mazzucato et al. 2008b), very recent analysis of segregating progenies would indicate that the two loci are not actually allelic.

sticky peel (pe)

see “Fruit skin” mutants Sundwarf mutants

Agostino Falavigna (CRA – Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) Franco Todisco (Università Cattolica del Sacro Cuore di Piacenza, Italy) Searching for a tomato plant with reduced, compact habit of possible advantage for open field cultivation and mechanical harvesting, Soressi’s attention was drawn by the so called “bird nest” phenotype. The name springs from the plant brachitism with curved leaves encircling the fruits like in a nest. The first “bird nest” variety, with hp, sp, u genes, is “Ottawa 60” released in Canada in 1964 by Loyd Lyall.

An sl-2 mutant anther withectopic ovules andcarpelloid structures.


By considering the existence of the “sundwarf” (sd) mutant (described by Rick in 1956) reduced in plant size, with internode progressively reduced in relation to light intensity and soil conditions, and its phenocopies, induced

by EMS and γ-rays respectively in S. Marzano and MoneyMaker, a series of crosses involving sundwarf mutants, two birdnest types and lines and determinate (sp) and indeterminate (sp+) cultivars was planned. The numerous F2 progenies were grown in open field for a not

easy and time consuming classification into distinct phenotypic groups (Todisco 1978). Notwithstanding the wide variation in plant habits ranging from indeterminate normal type to a rosette-like one (Fig. 1), the gathered data revealed the existence of two sundwarf genes (sd-2, incompletely recessive, and sd-3), interacting each other and with the sd, sp and br Mendelian factors. In addition, it was possible to recognize the birdnest phenotypes of cv. Ottawa 60 and H-105 from Canada as due to sd-2, sp and br homozygous genes (Falavigna and Soressi 1975, XIX SIGA Annual Congress; Falavigna and Soressi 1978). The field trial carried out with a sundwarf “cluster” phenotype (Fig. 2) at a population density of 130,000 plants/hectar evidenced a yield potential not significantly different from the best processing cultivars but earlier in fruit ripening (Soressi 1974; Falavigna and Soressi 1975; Soressi 1979). In later work at the Tuscia University in 2001, the pro5::uidA chimaeric reporter construct (see Lateral suppressor) was introduced in a sundwarf line for future studies on the IAA metabolism in relation to the plant habit influenced by different light spectrum and intensity.

Fig.1 Sundwarf tomato plants withextremely reduced habit (rosette).

Fig.2 Well performingsundwarf “cluster” tomatoplant in open field.


Tetranychus urticae Marisa Castagnoli CRA-ABP (ex Istituto Sperimentale per la Zoologia Agraria) Riccardo Caccia (Università della Tuscia, Viterbo Italy – presently different address) A critical aspect dealing with the use of transgenic plants is the global evaluation of their environmental impact. Tomato, because an economical important crop, and Tetranychus urticae (Acari Tetranychidae), a very injurious polyphagous mite, are the case study chosen to investigate unpredictable and undesirable effects on phytophagous arthropods. Three tomato near isogenic lines, i.e. the cv. Riogrande (RIG) and the transgenic lines RC332 [containing the Gox gene (see Glucose oxidase) and showing high glucose oxidase activity], and MS498 [containing the KTI3 gene (see Helicoverpa armigera) and exhibiting a high trypsin inhibition] were considered. Trichomes and contents of C and N of the leaves, differences in development and oviposition of T. urticae and damage caused were evaluated for each line. Laboratory trials evidenced that 1) the intrinsic rate of increase of two strains of T. urticae (T tomato-adapted strain, B non-tomato adapted strain), was significantly lower in RIG than in transgenic lines and doubling time ranged between 6.9 and 11.6 days in the first and between 3.9 and 5.3 days in the latter; 2) the glandular four-lobed trichomes were always higher in RIG than in other genotypes, 3) the N leaf content was from 1.3 to 1.9 fold lower and the C/N ratio from 1.3 to 1.9 fold higher in RIG than in other lines. Greenhouse experiments performed with an initially equal infestation of strain T evidenced: 1) no significant difference between plant lines in the final mite infestation (motile stages per plant), nevertheless an almost double number of spider mites was counted in RC332; 2) a significantly higher percentage of damaged leaves and a significantly higher average damage index on RC332 than on RIG (79% and 2.3 in the former, and 62% and 2.1 in the latter, respectively), even if in both transgenics a higher level of the most severe damages and a shorter time to approach them were observed; 4) a comparable number of mites causing the same damage level in all genotypes and a strong linear relationship between the first four levels of damage and mite infestation. Although in the laboratory studies both transgenic lines enhanced the T. urticae population increase, the glasshouse studies were not as conclusive and they only suggested the possibility of real differences between the transgenic and non-transgenic genotypes (Castagnoli et al. 2003).


Tomato, “fruit colour genes” Andrea Mazzucato, Maurizio Enea Picarella, Pietro Mosconi (Università della Tuscia, Viterbo Italy) The collection of a wide variety of germplasm entries and the selection of new mutant types and alleles allowed to work at the breeding of materials characterized by different colours of the fruit epidermis and flesh and, as such, by different contents of nutritional compounds. This included mutants and genes with altered composition of carotenoids (red, Beta, Delta), anthocyanins (Aubergine, Anthocyanin fruit, atroviolaceum) and chlorophyll (uniform, green flesh), with increased pigment composition (High pigment-1 and –2, Intense pigmentation) or lacking a fraction of pigments (colorless epidermis). A very first mention about the potential of this mutations for diversifying the appearance and the composition of the tomato products was given in early contributions (Soressi 1969b; 1969c). Later, using the germplasm retrieved in foreign collections from Butler and Darby, Soressi started to introgress genes and mutations for “colour genes” in the most important genetic backgrounds at that time (Marmande, San Marzano, Roma, New Yorker and Gimar). Such set of near isogenic lines has been partially lost; only the Gimar and San Marzano series remain active in Soressi’s collection. This breeding lines served as the basis for research aimed at establishing the pattern of pigment composition and

antioxidant activity in different backgrounds (Minoggio et al. 2003) Later, the study of tomato nutritional compounds has been undertaken using a comparison of the spectral characterisation of the tomato fruit surface pigments from the immature to over-ripe stage, using spectroscopy techniques based on visible fluorescence emission upon excitation in the same or ultraviolet spectral regions with the aim of verifying the spectral band for optimal conditions for fruit harvesting using non-destructive techniques. The main fluorescence spectral features belonging to

red, high-pigment-2 and sun black (sb); the last is a recent result of Soressi’sbreeding with “color genes”.


anthocyanins, flavonoids, carotenoids and chlorophyll a after excitation of skin tomato pigments at different laser wavelengths was identified (Lai et al. 2007). Another study investigated the combined effect of diluted seawater and ripening on the beneficial nutritional properties of tomato fruits from an antioxidant point of view. The research was conducted on tomato cv. Jama, Gimar wild type, Gimar gf, and Gimar nor. The genotypes Jama and Gimar gf showed increases in ascorbic acid, lipoic acid, and R-tocopherol during both ripening and salt treatment, whereas total ascorbate and tocopherols decreased in the berries from salt-treated plants of Gimar wild type. Ripening also determined decreases in ascorbate and tocopherol amounts in the Gimar nor line where a positive effect of ripening and salinity was observed (Sgherri et al. 2007). The research activity on “colour genes” has yielded collaborations with private companies (Barilla SpA, Tomato Colors) and initiatives for divulgation as a “bruschetta party” made with tomatoes of different colours (Mazzucato et al. 2007; Mazzucato and Picarella 2007).

Tomato, general Andrea Mazzucato, Maurizio Enea Picarella, Pietro Mosconi (Università della Tuscia, Viterbo Italy) The long and wide research activity resulted in a deal of review articles and book chapters or books focussed on the tomato crop (Soressi 1969a; 1969b; 1974; 1976; 1979; 1992; Porcelli et al. 1979; Amadei et al. 1990; Acciarri et al. 1998; Monti et al. 2004). Such research activity has allowed to put together a collection of tomato germplasm listing more than 650 accessions, that includes landraces, vintage and modern cultivars, genetic stocks and several other materials that, during the years, have undergone considerable characterisation. Compatibly with the absence of dedicated resouces, the collection has been maintained active, used for the internal research activity and distributed worldwide. Recently the collection has been transferred to a database in Microsoft Access format and more than 500 accessions have recently been included in the ECPGR Tomato Database (http://documents.plant.wur.nl/cgn/pgr/tomato/). The more recent activity of the Soressi and Mazzucato group at the Tuscia University has been described in a dedicated web site (http://www.unitus.it/tomato/).


Tomato, genetic markers Andrea Mazzucato (Università della Tuscia, Viterbo Italy) The use of natural or induced morphological variants, recognizable at an early stage of plant development, has been proposed as an aid to the production of hybrid tomato seeds under three different strategies (Soressi 1966; 1977; Sozzi 1969). Visible markers linked to male sterility genes were proposed in the perspective of maintaining genic male sterile parental lines, through early recognition of male sterile segregant; this is the case of anthocyanin absent (aa) and white virescent (wv) linked to male sterile-10 (ms-10, alias ms-35) and of anthocyianin without (aw) liked to ms-15 (Soressi 1974). Genetic markers expressed in the seed have been proposed to distinguish and/or select seeds from selfing or natural (NCP) or hand-outcrossing. Such

markers may involve the seed colour, such as brown seed inherited as an endosperm trait (bs, Soressi 1967a; Bin and Soressi, 1973; Soressi et al. 1975) or the seed coat structure, such as spongy seed inherited as a maternal trait (ss, Soressi 1967b) or both (Soressi 1968). Further, schemes using two, non allelic mutations for the bs character have been described (Soressi 1970b). The bs marker, together with those expressed in the plantlet, such as

potato leaf (c), may also serve in assessing unwanted crosses in the multiplication of seed parent lines with incomplete or conditional male sterility.

Tomato, high temperature tolerance Venkata Rami Reddy Sanampudi (Università della Tuscia, Viterbo Italy) The Intergovernmental panel on climate change (IPCC, 2001) announced that the global atmospheric temperature may raise 2.6°C by 2050 relative to 1990 and 5.8°C by 2010.Atmospheric temperature is the main important factor determining plant growth and productivity, hence many researchers are investigating high temperature effects on plant productivity. Alteration of the unsaturation levels of chloroplast membrane fatty acids could be a

A Soressi’s scheme for hybrid seedproduction aided by two, non allelicmutations for the bs character(Soressi 1970b).


useful method for enhancing the tolerance of plants to environmental stresses especially temperatures (Somerville and Browse 1991). Transgenic tobacco plants low in trienoic fatty acids exhibited high temperature

tolerance (Murakami et al. 2000). Plastid omega-3 fatty acid desaturase catalyzes the conversion of the dienoic fatty acids to trienoic fatty acids in glycerolipids which are the main components of chloroplast membranes of plants. At the University of Tuscia, we produced transgenic tomato plants that express double-stranded RNA (dsRNA) of tomato plastid omega-3 fatty acid desaturase gene which results in silencing of plastid omega-3 fatty acid desaturase gene.

Transformation was confirmed by PCR. Now we got first generation of transformed plants and we are going to test against high temperature and membrane fatty acids analysis.

Tomato hybrids, bs-marker based Giò Batta Bruzzone, Pasquale Martiniello, Agostino Falavigna (CRA – Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) By omitting the use of the male sterile genes (see stamenless-2; positional

sterile-2) we report briefly, the research activity, carried out under Soressi’s guide, focused on the practical utilization of the bs seed marker, in male fertile seed parent lines, which allows to directly recognize the F1 straw coloured seeds (+/bs) among the brown (bs/bs) ones (see Genetic markers).

Transformed tomato plantat flowering stage.

Fig.1 A bombus,one of the mostNCP responsibleinsect in tomato.

Fig.2 Simple vibratingdevice for improvingpollen recovery frommale parent plants.

Fig.3 Plant ofthe “Brown-1”tomato hybridobtained withbs/bs seedparent.


This knowledge has led to the development of theoretical schemes aimed at exploiting the Natural Cross Pollination (NCP) by pronubous insects (Fig. 1) and an optico-mechanical separation of the differently coloured seeds. Due to the lack of an efficient and economically convenient electronic equipment for F1 seed separation, Soressi’s attention was drawn on the technique based on hand pollination of bs/bs autofertile seed parent lines without flower emasculation (Bruzzone 1980; Falavigna et al. 1981). Such a procedure allows to obtain 99-100% F1 seeds in relation with stigma receptivity and pollen amount (Fig. 2). The synthesis of tomato hybrids (+/bs) of agronomic and commercial value requires a skilfull work for incorporating, through cross and backcross, the bs trait into the seed parent line that shall be combined with a pollen donor with normal seed colour. To choose the two parent lines, a previous work for evaluating the General and Specific Combining Ability, in relation with the final product (table or processing tomato), needs to be carried out (Maggiore et al. 1976; Martiniello and Soressi 1977). As the bs is present in homozygous situation in the seed parent lines and in heterozygous one in the F1 hybrids, the possible marker’s effect on plant and fruit characteristics was ascertained. Notwithstanding the delay in germination, initial seedling growth, flowering and fruit ripening of the bs/bs lines, no drawback is expected from using the bs gene, as marker, in the seed parent . In the 1977-79 triennium, a high number of F1 hybrids obtained by hand pollinating bs and bs-2 seed parent lines were included and evaluated in an outdoor comparison trials located in Northern, Central and Southern Italy. One of the best of these hybrids was inscribed, in 1982, in the National Register of Varieties with the name “Brown-1” (Fig. 3).

Tomato, hybrids for mechanical harvesting Carlo Lorenzoni (Università Cattolica del Sacro Cuore, Piacenza Italy) Tommaso Maggiore (Università di Milano, Italy) The possibility of the practical exploitation of hybrid vigour in processing tomato was studied in a period (early ‘70s) when attention at this regard was given mostly to determinate growth types. Two series of hybrids were produced, following a NC2 mating scheme (common mothers-common fathers) with 106 x 4 and 86 x 3 entries, respectively, where nearly all the parents were short vined (homozygous for the self pruning, sp, allele) varieties. The data, collected in two locations (Northern and Southern Italy), allowed an appreciation of the general combining ability of the single varieties for the characters under observation, i.e. flowering and first ripening time, time of picking, total and marketable yield, number of marketable fruits, percent of scrapped fruits, average fruit weight, ratio


between fruit diameters, degree of sun scald, degree of cracking, fruit firmness, soluble solid content (Maggiore et al. 1973; Soressi et al. 1973; Salamini et al. 1975; Maggiore et al. 1976; Conti et al. 1991). A comprehensive evaluation of the results presented and discussed let also draw some general conclusions: - the hybrids are usually earlier than the varieties; - as for the mean yield, the hybrids exceed the varieties by 20%, even though in the best environment some varieties can reach the highest level of production; when cultivated in suboptimal conditions, the F1s always overcome the varieties; - the hybrids seem to possess a higher phenotypic stability than the varieties; - the hybrids are clearly superior with regard to the fruit number per plant, but their fruit size is lower; it must be emphasized that high yielding plants are characterized by small or intermediate fruit size; -the fruit quality is not influenced by the hybrid vigour. High yielding F1s can be developed, by exploiting the genotypes explored: these hybrids should bear 40-60 fruits possessing a length/width ratio of 1.2, a weight of 50-60 g, and good fruit firmness. In a preliminary phase of the research, the efficiency in hybrid seed production was investigated in relation to variables such as plant genotype, location, frequency of pollinating insects, planting design, natural and hand pollination with and without emasculation. The results suggested procedures, combining genetic agronomic and entomological factors able to increase natural cross pollination rates avoiding hand work (Soressi and Lorenzoni 1970; Bin and Soressi 1973). Also suggestions for the use of male sterilty were thoroughly considered (Badino and Soressi 1976, see Male sterility; see stamenless-2).

Tomato, “high-pigment genes” Maurizio Enea Picarella, Enrico Santangelo (Università della Tuscia, Viterbo Italy) Antonella Lai (ENEA- Frascati, Roma Italy) Alberto Pardossi (Università di Pisa, Italy) In the middle of the 70’s, within Soressi’s breeding activity on fruit colour genes to be introduced in different backgrounds, special attention was driven to some mutants collectively referred to as “high-pigment group”. In particular, high pigment (hp), high pigment-2 (hp-2) and green flesh (gf) BC1S4 San Marzano breeding lines were studied. Whilst hp and gf had been provided by prof. Leonard Butler (see Tomato, “fruit colour genes”), hp-2, more recently isolated by C. Bowler (Mustilli et al. 1999), had been induced by EMS seed treatment in cv. Garim, a S. Marzano type (Soressi


1975). These monomendelian, recessive genes were attractive for enhancing leaf and fruit pigments as proved by spectrophotometric and HPLC measurements of chlorophyll a and b and carotenoids (Palmieri et al. 1975; 1978). To better clarify the effects of the “hp genes” on the mechanism of pigment production in leaf and in fruit ripening, their antioxidant value, chlorophyll a and b, total carotenoids and anthocyanins content were determined by spectrophotometric analyses. The assessed

relationship between the pigment content in the leaf and the fruit at different ripening stages led to the opportunity of adopting a more reliable method of analysis (Santangelo et al. 2002). For this reason total polyphenols, flavonols, flavonones, phenolic acid, lycopene and β-carotene

were determined in different tomato mutant lines and cultivars through mass-spectrometry and HPLC analyses; total antioxidant activity was also

measured. Opposite to a reasonable increase of carotenoids in the fruit of hp and hp-2, the level of flavonoids and other phenolics was lower than in the WT; reversely, it increased in mutants [lutescent-2 (l-2), rin/+, u] defective in carotenoids accumulation in the fruit (Minoggio et al. 2003). The biological activities of polyphenols are well known and today they are more appreciated for their beneficial effects on human health. Thus, when defining the total antioxidant activity of fruit and vegetables, polyphenols, besides carotenoid concentrations, should be considered as well. Because in the gf mature fruit some of the chloroplasts are not transformed into chromoplasts, this

mutant was judged particularly suitable in revealing possible levels of lipoic acid (LA), one of the most efficient biological antioxidants that mostly

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Fig.2. Macroscopic effect ofcontinuous light stress on tomatoleaves of green flesh (1), lutescent-2 (2), double recessive (gf, l-2) (3) and WT (4) NILs.

a cb

Fig.1. Different fruit pigmentation at thebreaking stage of S. Marzano NILshomozygous respectively for gf (a), gf, l-2 (b)and l-2 (c).


occurrs in green plant organs. In relation also to this issue, the capacity of salinity stress to increase the nutritional value of tomato (see Salt tolerance), was investigated in the commercial table tomato F1 hybrid Jama and in Gimar NILs (Gimar-WT, Gimar-nor, and Gimar-gf) grown in closed-loop rockwool culture. A striking result was the increase of the antioxidants ascorbic acid, tocopherols (α and γ), and to a minor extent of LA, in Gimar-gf fruits in comparison to Gimar-WT and Jama when plants were exposed to moderate salinity conditions (Sgherri et al. 2007). Now, it is known that hp-1 and hp-2 are isophenotypic mutations with exaggerated light responsiveness. The Hp-1 locus is the tomato ortholog of the gene encoding UV-damaged DNA Binding Protein 1 (DB1) and the phenotype of hp-2 is caused by alteration of DEETIOLATED-1 tomato ortholog. Both are responsible for enhancing the expression of genes involved in the production of fruit pigments via an interaction with chromatin. The gf mutation displays a non-functional stay-green phenotype with delayed chlorophyll degradation in the leaf during senescence and in the fruit during ripening. Efforts are being put forward in the Laboratory of vegetable biotechnology at the Tuscia University aiming to identify the lesion responsible for the gf phenotype. These are carried on by tools such as genetic analysis, microsynteny, physical mapping, functional analysis of candidate genes. Of particular interest has appeared the combination of gf with l-2, a mutant accelerating chlorophyll degradation. In double recessive plants, the presence of gf reduces the progressive de-pigmentation (yellowing) of the green parts, well noticeable in both l-2 leaf and fruit (Fig. 1). Such kind of interaction works also at the physiological level in a beneficial way for plant stress response such as continuous light exposure (Fig. 2). Physiological analyses on mutant lines with a role in plant senescence, like lesion mimic, and in fruit pigmentation enhancing (hp-1, hp-2), combined with gf and l-2, are also in the pipeline.

Tomato, leaf peroxidases

Gian Piero Soressi (Università della Tuscia, Viterbo Italy) The leaf peroxidase activity and the electrophoretic banding pattern of 69 tomato mutants affecting plant morphology have been studied (Soressi et al. 1974). The zymograms of 63 mutants were normal, whereas six mutants (lyrate, mottled, olivacea, monstruosa, extreme dwarf and Curl) were characterized by one or two more intensely stained bands. Their total peroxidase activity was significantly increased reaching the maximum in olivacea (40-folds higher than the WT). It could be inferred that the six abovementioned mutations influence directly or indirectly the peroxidase


activity level without affecting the genes which code for the peroxidase isozymes themselves. Parallels between the mutant effects on specific peroxidase bands and hormonally-mediated control of the peroxidase isozymes are discussed. Later, the activity of IAA oxidase in the mutants monstruosa and olivacea were studied (Palmieri et al. 1978). Analyses of the inheritance of the peroxidase activity and relative electrophoretic isoenzyme bands were olds done in crosses and backcrosses involving L. esculentum mutants and cvs with L. hirsutum accessions (Giovinazzi 1978). More recently, peroxidase activity was studied as a player in cell death responses controlled by the Pto-Fen complex following fenthion treatment (De Biasi et al. 2003, see Fen gene).

Triploidy

see Polyploidy uniform (u)

see Tomato, “high-pigment genes” Waste heat utilization in agriculture

Massimo Schiavi (CRA- Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi Italy) Tommaso Maggiore (Università di Milano, Italy) When Soressi was head of the ISPORT Montanaso Lombardo (Lodi) out Section, CTN-ENEL, Milan planned (1977) and built (1978-1980) an

experimental agro-thermic pilot plant at Tavazzano-Montanaso power station (Fig. 1) availing itself of the scientific and technical collaboration of the ISPORT Section Staff, in this case under M. Schiavi main responsibility and G.P. Soressi and T. Maggiore coordination. The experimental activity, aimed to prove technical and economic feasibility of using waste water heat

energy from the abovementioned station for crop production, was developed in two phases: an optimization term for identifying technical parameters to exploit at the highest level the relatively low (∆t=8°-9°C)

Fig.1 Overview of the experimental agro-thermic pilot plant at Tavazzano-Montanaso power station.


thermic gradient available; a second term for managing the crop cultivation trials for achieving the yield and related economic parameters. To obtain earlier crop production without negatively affecting marketable yield quantity and quality, the following experiments in plastic tunnels or open air were scheduled in relation to the different tested crops (Soressi et al. 1981): • Asparagus (green and white spears): in 3 tunnels with “EVA” plastic

cover; 1 unheated and 2 heated with waste water in 4 polyethylene (PE) pipes (2 buried in the soil under the crowns and 2 over soil (8-10 cm) in open air plot, unheated or heated in the same way.

• Strawberry: “EVA tunnels” (1 unheated, 1 heated in both soil and air). • Melon: “EVA tunnels” (1 unheated, 1 heated soil and air). • Lettuce: for 3 culture cycles (September-April) in 3 glasshouses, 1

unheated, 1 heated in both soil and air and 1 heated in soil, air and with heat water running on the roofs.

• Maize: late-ripening pure lines to be used as parents in F1 hybrid seed production, to get earlier yield seeds (heated and unheated big-plot in open air).

• Germinating cereal seedlings: barley, maize and wheat for integrating diary animal forage.

The best results putting in value the energy waste were obtained with maize pure lines, asparagus and strawberry. Regarding maize, the flowering was 5-8 days anticipated with a yield increment (up to 80%); in asparagus a significant earliness in spear emission and yield in both tunnels (26 days) and open air (18 days) was reached; as to the strawberry, the latest varieties

gave an earlier (20 days) yield increase (11%) (Schiavi et al. 1983; Schiavi et al. 1984b; 1984c; Rotino et al. 1985). By adopting a heat pump in 1983/84, a new trial for evaluating yield of Gerbera cut-flowers was set up in two glass-houses where the cv. Boston performed as the best in the autumn-winter period (Fig. 2) (Schiavi et al. 1984a). The possible advantages connected with the utilization of the water

low thermal level from electric power-plant are strictly dependent on the economic convenience of some technical actions aiming to avoid heat loss, rationalize the heat flux in relation to luminosity variations and to the cultivation techniques. Parallely, an aid system is recommended because

Fig.2 Gerbera cultivation in glasshouse,aerothermically heated through 20°Cwater circulation.


the needed continuous availability of hot water is not guaranteed being dependent on the working cycles of the electric power-plant (Soressi et al. 1985). In the subsequent decennium (1986-96) such a synergistic cooperation, between ISPORT section and CTN-ENEL at Tavazzano-Montanaso electric power plant, prosecuted by carrying out trials with Gerbera, leafy ornamentals, pepper, eggplant and tomato by adopting different soilless culture techniques.

Watermelon, germplasm evaluation Carlo Lorenzoni (Università Cattolica del Sacro Cuore, Piacenza Italy) Basilio Borghi (Parco Tecnologico Padano, Lodi Italy) At the beginning of the years ‘70s the observation that new cultivars of watermelon (Citrullus vulgaris L. Schrad) of foreign origin were substituting the local varieties on the Italian market, evidenced the need of an evaluation of the traditional germplasm as a source for breeding programmes. A collection of 43 cvs coming from Italy, Japan, Russia and USA was explored in order to: 1) obtain information about the inheritance of a series of characters (earliness of flowering and ripening; fruits per plant; firmness of rind; rind thickness / transversal diameter of the fruit ratio; sugar content of the ripe fruit juice; number and weight of the seeds per fruit; 1000 seeds weight); 2) evaluate the single cultivars as source of useful traits for the breeding; 3) estimate the possible advantages of the hybrids in comparison with the varieties; 4) put the basis for future breeding programmes (Bergamaschi 1971). The varieties and 22 hybrids, having at least one Italian variety as a parent, were compared in the field. In the trials, Italian varieties appeared outstanding as donors of useful characters for fruit setting and total yield, likely connected with a good environmental adaptation; USA entries were notable for yield and sugar content; the Japanese ones emerged for earliness, sugar content and low seed number; both Italian and Russian cvs resulted good sources for rind firmness. Crosses between Italian and imported genotypes seemed highly promising for selection in the segregating generation. On the contrary, the hybrids per se rarely demonstrated of value for a direct exploitation in cultivation. That does not exclude the possibility of individuating cross combinations convenient for the crop, but likely specific and wider programmes are required in order to find them (Soressi et al. 1974). For hybrid seed production technique in the cucurbitaceae, also the use of Ethrel as a phytoregulator able to suppress male flowers was evaluated (Borghi et al. 1972).


waxy fruit (pe-2) see “Fruit skin” mutants

Xanthomonas (resistance to) see “Lesion mimic” mutants


BIBLIOGRAPHY Acciarri N, Di Candilo M, Sanguineti MC, Soressi GP, (1998) Tomato. In: Italian

Contribution to Plant Genetics and Breeding, G.T. Scarascia Mugnozza & M.A. Pagnotta (eds.), pp. 463-465. University of Tuscia, Viterbo

Allavena A (1975) Induzione di forme polipoidi in L. esculentum Mill. Omozigote per il fattore pat di partenocarpia. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Allavena A, Falavigna A, Soressi GP (1976) Forme polipoidi partenocarpiche artificialmente indotte in Lycopersicon esculentum Mill. Colture Protette V(8-9): 45-46

Allavena A, Soressi GP (1978) Tetraploid parthenocarpic fruitful tomatoes. Tomato Gen Coop Rep 28: 2

Allavena A, Lancelotti M, Palmieri S, Soressi GP (1978) Influenza del gene nor2 su alcune caratteristiche qualitative della bacca di pomodoro durante la maturazione e la post-maturazione della pianta. I Conv. Naz. “Problemi e prospettive del pomodoro da mensa”, Monsampolo del Tronto, 7-8/11/1978 I.SP.ORT., Salerno, 145-164

Allavena A, Soressi GP (1982) Inclusione di tolleranza e/o resistenza alla batteriosi ad alone ed al virus del mosaico comune in fagiolo da granella (Phaseolus vulgaris L.). La Difesa delle Piante 5-6: 405-41

Allavena A, Fadda A, Soressi GP (1982a) New dry bean (Phaseolus vulgaris L.) varieties resistano to BCMV. Genet. Agr. 37: 138-139

Allavena A, Del Zan F, Fadda A, Murgut G, Siviero P, Tonetti T, Soressi GP (1982b) Attività di costituzione di varietà di fagiolo nano resistenti a BCMV e a Pseudomonas phaseolicola. Informatore Agrario 38 : 20625-20639

Allavena A, Fadda A, Soressi GP (1985a) "Niveo” e “Montebianco”: varietà di fagiolo (Phaseolus vulgaris L.) a seme bianco resistenti al virus del mosaico comune (BCMV). Sementi Elette, 31: 39-40

Allavena A, Fadda A, Soressi GP (1985b) “Montalbano” e “Mogano” varietà di fagiolo da granella (Phaseolus vulgaris L.) resistenti al virus del mosaico comune ottenute a seguito di trattamenti con MSE. Sementi Elette, 31: 37-38

Allavena A, Fadda A, Soressi GP, Salamini F (1986) Grazia and Patrizia bush bean. Hortscience 2: 1081-1082

Allavena A, Fadda A, Soressi GP (1989) “Montecarlo” and “Monterosa”, “Borlotto”-type dry beans. Hortscience 24: 1047-48

Allavena A, Angelini R, Genga AM, Pernacchia G and Soressi GP (1994) Advanced bean tissue culture and transformation research in Italy. Proceedings BARN NETWORK- CIAT, Cali, Colombia, 7-10 Settembre, 1993: pp 190-198

Amadei G, Trentini L, Soressi GP (1990) Il pomodoro – The tomato. Enichem agricoltura e Agrimont, Ed. unica Milano pp X + 91

Angelini Rota R (1987) Morfogenesi in vitro da tessuti somatici di species del genere Phaseolus. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Antognoni F, Ghetti F, Mazzucato A, Franceschetti M, Bagni N (2002) Polyamine pattern during flower development in the parthenocarpic fruit (pat) mutant of tomato. Physiol Plant 116: 539-547


Antonelli MM, Ferrari G, Portesi A, Fogher C, Soressi GP (1987) Plant regeneration in L. esculentum UC-82 and torosa-2 genotypes. Eucarpia Tomato Working Group, 2-6 September 1987, Salerno-ISPORT.

Antonelli M, Di Baccio D, Ederli L, Francini A, Marabottini R, Pellegrini E, Ciaffi M, Lorenzini G, Nali C, Pasqualini S, Santangelo E, Sebastiani L, Soressi GP, Badiani M (2005) Ozone as a tool for studying stress responses in plants: signalling and defence in normal, mutant and transgenic tomato lines. Proceedings XV Meeting of the Eucarpia Tomato Working Group, Bari, 20-23 September 2005. In press in Acta Horticulturae

Antonelli M, Marabottini R, Santangelo E, Francini A, Pellegrini E, Di Baccio D, Ederli L, Nali C, Pasqualini S, Soressi GP, Ciuffi M, Lorenzini G, Sebastiani L, Badiani M (2007) Ozone as a tool for studying stress responses in tomato (Solanum lycopersicum L.). III. Ethylene, cyanide and the development of foliar symptoms in the autonecrotic mutant V20368. In: Ramina A et al. (eds.) Advances in Plant Ethylene Research: Proceedings of 7th International Symposium on the Plant Hormone Ethylene (Pisa, 18-22 June 2006): 389-390

Astolfi S, Zuchi S, Picarella ME, Passera C, Soressi GP (2005) Salinity induces changes in nitrogen metabolism in tomato plants differing in salt tolerance. In: Plant nutrition for food security, human healt and environmental protection. C.J. LLi et al., (Eds.), 2005. Tsinghua University Press. Beijing, China; pp. 576-577

Badino M (1976) Il mutante di L. esculentum Mill. "short anthers": Eredità e sua utilizzazione del breeding. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Badino M, Soressi GP (1976) La maschiosterilità nella produzione di ibridi di pomodoro per la serra. Colture Protette 5: 41-45

Balestra GM, Caccia R, Antonelli MG, Soressi GP, Varvaro L (2000) Population dynamics of Xanthomonas visicatoria and Pseudomonas syringae pv. tomato on tomato plants expressing specific (Pto) and aspecific (Gox) resistance genes. J Plant Pathol 82: 77

Baroni D (1994) Confronto tra genotipi di pomodoro differenziati per portamento e partenocarpia in coltura anticipata di piena campo. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Bedini C, Caccia R, Triggiani D, Mazzucato A, Soressi GP, Tiezzi A. Micropropagation of Aloe arborescens Mill.; a step towards efficient production of its valuable leaf extracts. Submitted to Plant Biosystems

Beraldi D (2001) Ricerca di marcatori molecolari associati al gene di partenocarpia parthenocarpic fruit (pat) in pomodoro. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. A Mazzucato

Beraldi D, Picarella ME, Soressi GP, Mazzucato A (2004) Fine mapping of the parthenocarpic fruit (pat) mutation in tomato. Theor Appl Genet 108: 209-216

Bergamaschi G (1971) Valutazione di cultivar di anguria e di loro incroci in funzione del miglioramento genetico. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Bianchi A, Marchesi G, Soressi GP (1963) Some results in radiogenetical experiments with tomato varieties. Rad Bot 3: 333-343

Bianchi A, Marchesi G, Soressi GP (1964) Alcuni risultati in esperimenti di radiogenetica con diverse cv. di pomodoro. Genetica Agraria 18: 317-318


Bianchi A, Soressi GP (1969) Mutanti di pomodoro artificialmente indotti suscettibili di utilizzazione nel miglioramento genetico. Sementi Elette XV (3), 2-6

Bianchi A, Borghi B, Lorenzoni C, Salamini F, Soressi GP (1973) Acquisizioni di genetica vegetale e insegnamento. Atti Ass Genet Ital 18:79-90

Bima PJ, Mensurati F, Soressi GP (1993) Rigenerazione da ipocotile in pomodoro (L. esculentum Mill.) su substrato privo di ormoni. Atti XXXVII Convegno Annuale SIGA, Orvieto, 11-14 Ottobre 1993:184 (abstract)

Bima PJ (1997) Studio dei fattori che condizionano la micropropagazione in diverse cultivar di aglio (Allium sativum L.). PhD Thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Bima PJ, Mensurati F, Soressi GP (1995) Effects of gibberellic acid (GA3) and micropropagation on axillary shoot induction in monostem genotype (to-2) of tomato. Current Issues in Plant Molecular and Cellular Biology. M Terzi, R Cella, A Falavigna (Eds). Kluwer Academic Publisher, London, pp 411-416

Bin F, Soressi GP (1973) Insetti pronubi e produzione di seme ibrido di pomodoro. Genetica Agraria 27: 35-74

Blanco A, Soressi GP (1988) Ibridazione interspecifica ed intergenerica. In: GT Scarascia Mugnozza (ed) Miglioramento genetico vegetale. Collana: Scienza e tecniche delle produzioni vegetali - Patron, Bologna. pp. 298-308

Borghi B, Maggiore T, Soressi GP, Poma Treccani C (1972) Tecnica di produzione di seme ibrido nelle cucurbitacee mediante l’impiego dell’Ethrel. Riv Orto Floro Frutticoltura Italiana 56: 750-759

Bricchi D (1993) Espressione del gene pat-2 e variazione di alcuni regolatori endogeni di crescita in due versioni quasi isogeniche – normale e partenocarpica – di un ibrido di pomodoro allevato in tunnel freddo. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Broadway RM, Duffey SS (1986) Plant proteinase inhibitor: mechanism of action and effect on the growth and digestive physiology of larval Heliothis zea and Spodoptera exigua. J Insect Physiol 32: 827-833

Bruzzone GB (1980) Una nuova tecnica per la produzione su larga scala di seme ibrido di pomodoro: impollinazione, senza emasculazione, di linee portaseme maschiofertili geneticamente marcate. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Bruzzone GB, Falavigna A, Soressi GP (1983) Metodologia per la sintesi di ibridi F1 di pomodoro mediante impiego del marcatore bs del seme. Giornate GOF-SOI, Ferrara, 9 December 1983, Informatore Agrario n.s.: 45-47.

Bruzzone G B, Carnevali G, Frangi P and Soressi GP (1984). Il pomodoro “monostelo”: stato attuale della ricerca. Notiziario tecnico. ERSO, 17, 7-13.

Caccia R (1998) Impiego del transgene codificante l’enzima glucosio-ossidasi (GOD) in pomodoro ai fini della comprensione e manipolazione della tolleranza a stress biotici. PhD thesis. Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi.

Caccia R, Delledonne M, Balestra MG, Varvaro L, Soressi GP (1999) Plant-bacterial pathogen interaction modified in transgenic tomato plants expressing the Gox gene encoding glucose oxidase. In "Genetics and Breeding for Crop Quality and Resistance”. Kluwer Academic Publisher, Netherlands, pp 119-125

Caccia R, Delledonne M, Levine A, De Pace C, Mazzucato A (2001) Apoptosis-like DNA fragmentation in leaves and floral organs precedes their developmental senescence. Plant Biosystems 135: 183-190


Cammareri G (2003). Rigenerazione e trasformazione del mutante lateral suppressor (ls) con un gene artificiale di partenocarpia (DefH9-iaaM) ed incremento dell’efficienza di micropropagazione in pomodoro (Lycopersicon esculentum Mill.). Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutors Proff. GP Soressi and A Tiezzi

Campion B (1977). Potenzialità del mutante torosa-2 nel miglioramento genetico del pomodoro per raccolta meccanica. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Campion B, Fadda A, Soressi GP (1981) Conversione a taglia nana del fagiolo di Spagna rampicante (P. coccineus L.). Proceedings of “Risultati di un quinquennio di lavoro sul miglioramento genetico del fagiolo da granella” Meeting, University of Turin, 3 July, Cuneo Italy

Campion B, Falavigna A, Soressi GP, Schiavi M (1985) Efforts for in vitro androgenesis in onion (Allium cepa L.). Genet Agr 39: 305

Campion B (1995) ‘Venere’ and ‘Alarico’, New Scarlet Runner Bean (Phaseolus coccineus L.) Cultivars with Determinate Growth Habit. HortScience 30: 1483-1484

Cantoni M (1991) Influenza del gene pat-2 su allegagione e caratteristiche produttive delle versioni quasi isogeniche normale e partenocarpica di un ibrido F1 di pomodoro allevato in tunnel freddo. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Caporali F, Soressi GP (1993) Ruolo dell'orticoltura nella prospettiva di un sistema agricolo integrato. Informatore Agrario 49 (7 suppl.): 86-88

Carrai C, Fioravanti L, Tomassini C, Serra G, Soressi GP (1990) Miglioramento genetico della Dieffenbachia. I. Tecnica di incrocio. Colture Protette 11: 107-114

Carrai C, Fioravanti L, Cavallini A, Serra G, Soressi GP (1993) Caratterizzazione citologica di cultivar di Dieffenbachia. Colture protette 2: 101- 104

Castagnoli M, Caccia R, Liguori M, Simoni S, Marinari S, Soressi GP (2003) Tomato transgenic lines and Tetranychus urticae: changes in plant suitability and susceptibility. Exp Appl Acarol 31:177-188

Castelnuovo M (1984) Studio di fattori che influenzano l’embriogenesi somatica e la micropropagazione in vitro del fagiolo. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Chiappari A (1993) Indagine sull’accumulo e la distribuzione della sostanza secca e alcune sue componenti in genotipi monostelo e normale di pomodoro (L. esculentum). Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Colla G, Rouphael Y, Cardarelli M, Mazzucato A, Olimpieri I (2007) Growth, yield and reproduction of dwarf tomato grown under simulated microgravity conditions. Plant Biosystems 141: 75-81

Conti S, Di Candilo M, Soressi GP, Sanguineti MC, Dadomo M, Del Re L, Pasotti PP (1991) Indagini su ibridi di pomodoro da industria. Sementi Elette 37(1): 9-16

Cravedi P (1966) Mutanti artificiali indotti in cultivar di pomodoro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Cravedi P, Soressi GP (1969) Mutagenic effects of EMS in tomato under different treatment conditions. Genetica Agraria 23: 235

Crinò P, Cristinzio G, Gentile A, Haegi A, Rugini E, Soressi GP (1996) Filtrati colturali e tossine nella selezione di piante resistenti a malattie. Petria 6 (suppl. 1):197-217


Cunico C (1991) Caratterizzazione di varianti somaclonali indotti in pomodoro (Lycopersicon esculentum) a seguito di rigenerazione in vitro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

De Biasi MG, Astolfi S, Acampora A, Zuchi S, Fonzo V, Santangelo E, Caccia R, Badiani M, Soressi GP (2003) A H2O2-forming peroxidase rather than a NAD(P)H-dependent O-

2 synthase may be the major player in cell death responses controlled by the Pto-Fen complex following Fenthion treatment. Funct Plant Biol 30: 409-17

De Logu G (1974) Partenocarpia in Lycopersicon esculentum Mill.: dati relativi a progenie F3 omozigoti per un fattore monogenico inducente lo sviluppo di bacche apirene. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Di Rollo B (1998) Trasformazione genetica del pomodoro mediante Agrobacterium tumefaciens utilizzando il gene AtCyS di Arabidopsis codificante una cistatina per indurre resistenza ad insetti dannosi. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Donati P (1990) Produzione di seme ibrido in cipolla mediante maschiosterilità citoplasmatica. Uso di gametocidi e tecnica seed-to-seed con valutazione tecnico-economica. Graduation Thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Falavigna A and Soressi GP (1978) Birdnest phenotypes as related with sundwarf genes. Tomato Genet Coop Rep 28: 6

Falavigna A, Badino M, Soressi GP (1978) Potential of the monomendelian factor pat in the tomato breeding for industry. Genetica Agraria 32, 159-160 (Abstr.)

Falavigna A, Badino M, Bruzzone GB, Soressi GP (1981) A new technique for tomato hybrid seed production: direct hand pollination of male firtile bs lines. Acta Horticulturae 111: 47-54.

Falavigna A, Roversi N, Soressi GP (1982) Brown-1: ibrido F1 di pomodoro da industria realizzato con linee portaseme bs. Informatore Agrario 20: 21017-18.

Falavigna A and Soressi GP (1980) Ibridi di pomodoro da industria: risultati conseguito nel triennio 1977-79 e prospettive. Atti Convegno su Miglioramento Genetico del pomodoro da industria, Sorrento Sant’Agnello, 8-9 November 1972. Arti Grafiche Del

Falavigna A, Soressi GP (1981a) Miglioramento genetico dell’asparago di Altedo: stato attuale delle ricerche. Seminario sulla coltura dell’asparago, Villa Salina (BO) 29 giugno 1979, pp. 9-15

Falavigna A, Soressi GP (1981b) La coltura in vitro di antere: una efficace tecnica per la sintesi di linee pure in Asparagus officinalis L. Genet Agr 35: 62-64

Falavigna A, Tacconi MG, Soressi GP (1983) Recent progress in asparagus (Asparagus officinalis L.) breeding by anther vitro culture. Acta Horticulturae 131: 215-222

Falavigna A, Casali PE, Mazzanti C, Soressi GP (1984a) Metodologia per la sintesi di ibridi tra cloni eterozigoti di asparago. Atti giornate GOF-SOI, 9 December 1983, Ferrara, Italy, pp. 20-21

Falavigna A, Tacconi MG, Casali PE, Soressi GP (1984b) Variability in asparagus (Asparagus officinalis L.) following anther in vitro culture. Genet Agr 38: 330-331

Falavigna A, Soressi GP (1987) Influence of the pat-sha gene on plant and fruit traits in tomato (L. esculentum Mill.). In: Modern trends in tomato genetics and breeding, p. 128. Proceedings of the 10th Meeting of the EUCARPIA Tomato Working Group, 2-6 Sept. 1987, Salerno, Italy


Falavigna A, Soressi GP (1993) Miglioramento genetico per la qualità di specie orticole. Informatore Agrario 49 (6): 61-65

Falavigna A (2007) Asparago: resa e qualità col miglioramento genetico. Agricoltura 19: 85-87 (Regione Emilia Romagna)

Falavigna A, Huaisong W, Alberti P, Casali PE, Mennella G, Toppino L (2008) Interspecific hybridization for asparagus breeding in Italy. Acta Horticulturae 776: 291-298

Fantino MG, Schiavi M (1987) Onion breeding for tolerance to Fusarium oxysporum f. sp. cepae, in Italy. Phytopathol Medit 26: 108-112

Fioravanti L, Carrai C, Tomassini C, Serra G, Soressi GP (1990) Miglioramento genetico della Dieffenbachia. II. Programmazione della fioritura mediante trattamenti con GA3. Colture Protette 11: 103-107

Frangi P (1984) Confronto per parametri di sviluppo e di produzione tra genotipi di pomodoro (L. esculentum Mill.) monostelo e normale. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Frangi P and Soressi GP (1986). Confronto per parametri di sviluppo tra genotipi monostelo e normale di pomodoro (Lycopersicon esculentum Mill.). Annali Facoltà di Agraria, Università Cattolica S. Cuore, Milano 26(1), 15-28.

Gandolfi G (1992) “Breeding lines” monostelo per la costituzione di varietà e ibridi di pomodoro da industria. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Gianfilippi F (2006) Studio della diversità molecolare in popolazioni locali italiane di lenticchia (Lens culinaris Medik) tramite marcatori ISSR. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. A Mazzucato

Giovinazzi F (1978) Controllo dell’attività perossidasica e delle bande in incroci di Lycopersicon esculentum Mill. e L. hirsutum E. Bompi. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Gorguet B, Schipper D, van Heusden AW, Lindhout P (2006) High-resolution Wne mapping of ps-2, a mutated gene conferring functional male sterility in tomato due to non-dehiscent anthers. Theor Appl Genet 113: 1437–1448

Gorguet B, Eggink PM, Ocaña J, Tiwari A, Schipper D, Finkers R, Visser RG, van Heusden AW ( 2008) Mapping and characterization of novel parthenocarpy QTLs in tomato. Theor Appl Genet 116:755-67

Habashy AA (1993) Miglioramento della tolleranza allo stress salino in pomodoro: variabilità' somaclonale indotta a seguito di rigenerazione in presenza di NaCl ed individuazione di parametri in vitro utili nella selezione. PhD Thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Habashy AA, Testa G, Mosconi P, Caccia R, Mazzucato A, Santangelo E, Soressi GP (2004) Parthenocarpy restores fruitfulness in sterile triploid (3x) tomatoes artificially obtained by crossing 4x × 2x somaclones. J Hort Sci Biotechnol 79: 322-328

IPCC (2001) Climate change 2001:impacts,adaptation and vulnerability technical summary (www.ipcc.ch)

Kagan-Zur V, Yaron-Miron D, Mizrahi Y (1991) A study of triploid tomato fruit attributes. J Am Soc Hort Sci 116: 228-31

Kiferle C (1998) Induzione di poliploidi partenocarpici in pomodoro mediante rigenerazione in vitro di espianti da ipocotile di genotipi pat e pat-2. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi


Lai A, Santangelo E, Soressi GP, Fantoni R (2007) Analysis of the main secondary metabolites in tomato (Lycopersicon esculentum Mill.) epicarp tissue during fruit ripening using fluorescence techniques. Postharvest Biol Technol 43: 335-342

Lancellotti M (1978) Effetti del gene “non ripening” (nor2) sulla qualità della bacca di pomodoro durante la maturazione. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Lioi L, Piergiovanni AR, Soressi GP, Nigro C, Tamietti G, Turina M, Campion B (2007) Caratterizzazione, selezione, risanamento e valutazione di cultivar locali di fagiolo comune (Phaseolus vulgaris L.). Italus Hortus 14: 31-40

Maggioni L, Soressi GP (1992) Risorse genetiche delle specie orticole e loro salvaguardia. pp. 64-97. In "Ambiente Italia". Lega per l'Ambiente pp. 501. Arancia blu. Vallecchi editore, Firenze

Maggiore T, Lorenzoni C, Soressi GP, Mariani BM, Salamini F (1972) Ibridi opaco-2 in Italia settentrionale. Maydica 17: 67-94

Maggiore T, Borghi B, Lorenzoni C, Mariani BM, Salamini F, Soressi GP (1973) Determinated hybrid tomatoes. I. Comparison between hybrids and varieties and definition of a standard fruit type for mechanical harvesting. Genet Agr 27: 235-280

Maggiore T, Motto M, Salamini F, Soressi GP (1976) Attitudine combinatoria per l’adattabilità alla coltura protetta in pomodoro (L. esculentum Mill.). Sementi Elette 22: 9-16

Mapelli S, Frova C, Torti G, Soressi GP (1978) Relationship between set, development and activities of growth regulators in tomato fruits. Plant Cell Physiol. 19: 1281-1288

Mapelli S, Torti G, Badino M, Soressi GP (1979). Effects of GA3 on flowering and fruit-set in a mutant of tomato. HortSci 14: 736-737

Mapelli S, Bricchi D, Cantoni M, Soressi GP (1995) Gene pat-2 e livelli di fitoregolatori endogeni, allegagione e caratteristiche produttive in pomodoro. Italus Hortus 2:74-79

Marchesi G, Soressi GP (1965) Risposta di due cultivar di pomodoro al trattamento con metansulfonato di etile. Atti Ass Genet It Pavia 10: 185-198

Marras R (2002) Analisi di espressione della mutazione di pomodoro parthenocarpic fruit-2, costituzione di una popolazione segregante F2 e fasi preliminari della ricerca di marcatori molecolari associati al locus. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. A Mazzucato

Martiniello P (1975) Effetti biochimici di alcuni geni “Never ripe” sulla bacca di pomodoro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor prof. GP Soressi

Martiniello P and Soressi GP (1977) Valutazione dell’attitudine combinatoria generale e specifica di linee bs di pomodoro. Annali ISPORT 8, pubbl. 138, pp.12.

Martiniello P, Falavigna A, Soressi GP (1985) Influence of the tomato (Lycopersicon esculentum Mill.) brown seed (bs) gene on plant and fruit characteristics. Genet. Agr. 39: 417-422.

Mazzucato A, Pepponi F, Chioccia G, Soressi GP (1998a) Ricostituzione e valorizzazione della varietà locale di pomodoro Scatolone di Bolsena. Italus Hortus 3: 30-35


Mazzucato A, Taddei AR, Soressi GP (1998b) The parthenocarpic fruit (pat) mutant of tomato (Lycopersicon esculentum Mill.) sets seedless fruits and has aberrant anther and ovule development. Development 125: 107-114

Mazzucato A, Caccia R, Soressi GP (1998c) Expression of the pat syndrome and other floral traits in a tomato BC1 population after interspecifìc cross with L. pennelli. Tomato Genet Coop Rep 48: 33-36

Mazzucato A, Testa G, Biancari T, Soressi GP (1999) Effect of gibberellic acid treatments, environmental conditions, and genetic background on the expression of the parthenocarpic fruit mutation in tomato. Protoplasma 208: 18-25

Mazzucato A, Olimpieri I, Ciampolini F, Cresti M, Soressi GP (2003) A defective pollen-pistil interaction contributes to hamper seed set in the parthenocarpic fruit tomato mutant. Sex Plant Reprod 16: 157-164

Mazzucato A, Mosconi P, Siligato F, Picarella ME, Soressi GP (2006) Caratterizzazione di varietà locali di pomodoro coltivate nella regione Lazio. Italus Hortus 13: 735-740

Mazzucato A, Olimpieri I, Rossi M, Caccia R, Soressi GP (2006) A new reporter construct to monitor IAA dynamics during tomato development. Tomato Genetics Cooperative Report 56: 26-28

Mazzucato A, Picarella ME (2007). Tomato bruschetta. SOL Newsletter 17: 10 Mazzucato A, Olimpieri I, Soressi GP (2007) Bruschette a colori? Sì, grazie. Il La 1: 74-

76 Mazzucato A, Papa R, Bitocchi E, Mosconi P, Nanni L, Negri V, Picarella ME, Siligato

F, Soressi GP, Tiranti B, Veronesi F (2008a) Genetic diversity, structure and marker-trait associations in a collection of Italian tomato (Solanum lycopersicum L.) landraces. Theor Appl Genet 116: 657-669

Mazzucato A, Olimpieri I, Siligato F, Picarella ME, Soressi GP (2008b) Characterization of genes controlling stamen identity and development in a parthenocarpic tomato mutant indicates a role for the DEFICIENS ortholog in the control of fruit set. Physiol Plant 132: 526–537

Mendoza de Gyves E (1995) Messa a punto e verifica di metodi di trasformazione in fagiolo (Phaseolus spp.). PhD Thesis, Università dellaTuscia, Viterbo, Italy. Tutor Prof GP Soressi

Mensurati F (1991) Selezione di piante di pomodoro insensibili a filtrato colturale di Phytophtora infestans in varietà parzialmente resistenti al fungo. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Dott. Paola Crinò

Mensurati F (1994) Induzione di variabilità in pomodoro tramite rigenerazione in vitro, mutagenesi chimica e loro combinazioni. PhD thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Minoggio M, Bramati L, Simonetti P, Gardana C, Iemoli L, Santangelo E, Mauri PL, Spigno P, Soressi GP, Pietta PG (2003) Polyphenol pattern and antioxidant activity of different tomato lines and cultivars. Ann Nutr Metab 47:64-69

Monti L, Santangelo E, Corrado G, Rao R, Soressi GP, Scarascia Mugnozza GT (2004) Il “San Marzano”: problematiche e prospettive in relazione alla sua salvaguardia e alla necessità di interventi genetici. Agroindustria 3: 161-169

Moretti F (1982) Valutazione di tecniche di produzione del seme di cipolla. Graduation Thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Motto M, Soressi GP, Salamini F (1975) Mutation frequencies and chimeric formation in Phaseolus vulgaris after EMS treatment of dormant seeds. Rad Bot 15: 291-29


Motto M, Soressi GP, Salamini F (1979a) Growth analysis in a reduced leaf mutant of common bean (Phaseolus vulgaris L.). Euphytica, 28: 593-600

Motto M, Fadda A, Soressi GP, Salamini F (1979b) Lisa P71: varietà nana di fagiolo da granella a seme bianco resistente al virus del mosaico comune (BCMV). Ann. ISPORT 8 (123): 1-5

Murakami Y, Tsuyama M, Kobayashi Y, Kodama H, Iba K (2000) Trienoic fatty acids and plant tolerance of high temperature. Science 287: 476–479

Mustilli AC, Fenzi F, Ciliento R, Alfano F, Bowler C (1999) Phenotype of the tomato high pigment-2 mutant is caused by a mutation in the tomato omolog of DEETIOLATED1. Plant Cell 11: 145–157

Odoardi M, Gentinetta E, Soressi GP, Salamini F (1976a) Metabolismo del saccarosio nel seme di fagiolo a diversi stadi dello sviluppo. Annali ISPORT, Salerno 4: 57-66

Odoardi M, Gentinetta E, Soressi GP, Salamini F (1976b) Forme multiple della fosfoesoisomerasi nel seme di fagiolo durante lo sviluppo. Annali ISPORT, Salerno 4: 69-71

Odoardi M, Palmieri S, Giacomozzi G, Soressi GP, Salamini F (1976c) Enzimi del metabolismo dei carboidrati durante lo sviluppo del seme in pisello liscio e rugoso. Riv. Agron. 10: 233-239

Olimpieri I (2006) Aspetti genetici e molecolari della partenocarpia in pomodoro. PhD thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Olimpieri I, Siligato F, Caccia R, Mariotti L, Ceccarelli N, Soressi GP, Mazzucato A. (2007) Tomato fruit set driven by pollination or by the parthenocarpic fruit allele are mediated by transcriptionally-regulated gibberellin biosynthesis. Planta 226: 877-888

Olimpieri I, Mazzucato A. Phenotypic and genetic characterization of the pistillate mutation in tomato. Submitted to Theor Appl Genet

Palmieri S, Martiniello P, Soressi GP (1975) Effetti di alcuni geni “Never Ripe” e “High Pigment” sui principali pigmenti della bacca di pomodoro (L. esculentum Mill.). Annali Istituto Sperimentale per l’Orticoltura, Salerno, VII: 1-12

Palmieri S, Sozzi A, Gorini FL, Soressi GP (1976) Influenza dei geni Nr e nor sulla qualità e conservabilità del pomodoro (L. esculentum Mill). Annali I.V.T.P.A., 6: 167-178 (oral comunication presented at Convegno Annuale SIGA, 9-11 October, 1975, Milan-Minoprio)

Palmieri S, Martiniello P, Soressi GP (1978) Chlorophyll and carotene content in high pigment and green flesh fruits. Tomato Genet Coop Rep 28: 10

Palmieri S, Soressi GP (1978) Effetti di alcuni geni Never ripe e high pigment su colorazione e valore vitaminico della bacca di pomodoro. I Conv. Naz. “Problemi e prospettive del pomodoro da mensa”, Monsampolo del Tronto, 7-8/11/1978 I.SP.ORT. Salerno, 95-99

Palmieri S, Odoardi M, Soressi GP, Salamini F (1978) Indoleacetic acid oxidase activity in to high peroxidase tomato mutants. Physiol Plant 42: 85-90

Paniconi G (2006) Caratterizzazione morfologica e molecolare di una varietà di fagiolo (Phaseolus vulgaris L.) di origine siciliana. Graduation Thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. A. Mazzucato

Pardossi A, Frangi P, Tognoni F, Soressi GP (1988) Growth analysis of monostem tomato genotype in N.F.T. Acta Hortic 229: 361-369

Parsons EP (2000) Sviluppo di un protocollo di rigenerazione in vitro ai fini della trasformazione genetica del fagiolo (Phaseolus coccineus L. e Phaseolus vulgaris


L.). Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof GP Soressi

Pecaut P, Philouze J (1978) A sha pat line obtained by natural mutation. Rep Tom Genet Coop 28: 12

Pepponi F (1995) Ricostituzione e valorizzazzione dell’ecotipo di pomodoro “Scatolone di Bolsena”. Graduation thesis, Univ. Tuscia, Viterbo. Tutor prof. GP Soressi

Perri E (1991). Effetto della composizione del substrato, del tipo di espianto e del genotipo sulla frequenza di rigenerazione e di variazione indotta in vitro in pomodoro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Philouze J (1985) Parthénocarpie naturelle chez la tomate. II. Etude d'une collection variétale. Agronomie 5: 47-54

Philouze J, Pecaut P (1986) Parthénocarpie naturelle chez la tomate. III Etude de la parthénocarpie due au gène pat (parthenocarpic fruit) de la lignée «Montfavet 191». Agronomie 6: 243-248

Picarella ME, Antonelli M, Astolfi S, Zuchi S, Vernieri P, Soressi GP (2007) Ethylene and ABA cross-communication and plant growth response to salt stress in tomato (Solanum lycopersicum L.). Advances in Plant Ethylene Research: Proceedings 7th International Symposium on the Plant Hormone Ethylene (Pisa, 18-22 June 2006): 113-114

Poma G (1992). Selezione di linee monostelo-partenocarpiche utili ai fini del miglioramento genetico del pomodoro da tavola e da industria. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Porcelli S, Soressi GP, Lunicini L (1979) Miglioramento genetico del pomodoro da mensa. Colture Protette 8: 19-24

Portesi A (1987) Analisi fenotipica e genetica di mutanti della buccia in pomodoro (L. esculentum Mill.). Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Portesi A, Frangi P, Soressi GP (1987) Allelism test between the fruit skin mutants p, nor3, pe and pe-2. Tomato Genet Coop Rep 37: 59

Portesi A, Soressi GP (1988) Analisi fenotipica e genetica di mutanti della buccia in pomodoro. Proceedings of the Symposium on “Physiology of fruit drop, ripening, storage and post-harvest processing of fruits”, Turin, Italy, pp. 128-135

Quattrini E (1989) Moltiplicazione vegetativa in vitro di cloni diplo-aploidi in asparago. Graduation thesis, Università Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Ranalli P, Parisi P, Soressi GP, Habashi AA, Santangelo E (2001) Fagiolo (Phaseolus vulgaris L.) in Leguminose e agricoltura sostenibile: specie da granella e cover crops. Calderoni Ed agricole, Bologna, pp. 447-502

Ranieri R, Stancanelli G, Tonelli A, Ricci A, Soressi GP (1996) Variabilità somaclonale in pomodoro da industria (L. esculentum Mill). Italus Hortus 3: 14-21

Remotti P (1990) Variabilità in popolazioni derivate da incrocio Solanum tuberosum L. x S. Berthaultii per caratteri morfologici e per densità di tricomi ghiandolari correlati con la resistenza agli insetti. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Ryan CA (1990) Proteinase inhibitors in plants: Genes for improving defenses against insect and pathogens. Annu Rev Phytopathol 28: 425-449


Rick CM, Uhlig JW, Jones AD (1994) High alpha-tomatine content in ripe fruit of Andean Lycopersicon esculentum var. cerasiforme: Developmental and genetic aspects. Proc Natl Acad Sci USA 91: 12877-12881

Rossi F (1977) Influenza dei geni “Never ripe” sulla conservabilità delle bacche di pomodoro (L. esculentum Mill.). Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Rossi M (2003) Caratterizzazione fenotipica e istologica di mutanti di pomodoro coinvolti nel metabolismo dell’auxina mediante costrutti reporter auxino-inducibili. PhD Thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Rotino GL, Schiavi M, Manera M, Roversi N, Casarotti D, Soressi GP (1985) Impiego di calore di acque residue da centrali termoelettriche nella coltura di cultivar di fragola neutro-diurna. Colture Protette 14: 51-56

Ruiu F (2006) Integrazione della mutazione positional sterile-2 (ps-2) nella mappa molecolare di pomodoro. Graduation Thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. A Mazzucato

Salamini F, Maggiore T, Lorenzoni C, Soressi GP (1975) Ibridi di pomodoro. III. Attitudine combinatoria di 86 varietà da industria. Sementi Elette 21: 3-23

Salamini F, Fadda A, Allavena A, Motto M, GP Soressi (1978) Nuove varietà di fagiolo resistenti al mosaico comune (BCMV) costituiti in Italia. Informatore Agrario 34 (50): 3990-3

Santangelo E (1988) Espressione del gene pat-2 di partenocarpia in pomodoro in condizioni di fluttuazioni termiche e suo effetto su precocità e stabilità della produzione. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Santangelo E, Soressi GP (1990) La partenocarpia nel pomodoro. Colture Protette 3: 29-33

Santangelo E, Benedettelli S, Tomassini C, Soressi GP (1990) Comportamento in tunnel freddo delle versioni, quasi isogeniche, normale e partenocarpica (pat-2) di tre ibridi di pomodoro. Colture Protette 11: 81-90

Santangelo E, Balestra GM, Varvaro L, Soressi GP (1998) Ottenimento di piante di pomodoro tolleranti sia a Fenthion che a Pseudomonas syringae pv. tomato a seguito di rigenerazione in vitro, in presenza di Fenthion, di espianti cotiledonari eterozigoti per il gene Pto di resistenza al batterio. Atti Giornate Fitopatologiche, Scicli e Ragusa 3-7 Maggio 1998, pp. 719-724

Santangelo E, Balestra GM, Varvaro L, Soressi GP (2000) Efficiency of the in vitro selection for R4 tomato plants resistant or susceptible to both fenthion and Pseudomonas syringae pv. tomato. J Plant Pathol 82: 83

Santangelo E, Lai A, Fonzo V, Mosconi P, Fantoni R, Soressi GP (2002) Studio dell’evoluzione di clorofilla, carotenoidi e polifenoli durante la maturazione della bacca in linee di pomodoro diversificate per geni del colore, mediante analisi ottiche. Proceed VI Giornate Scientifiche (vol. II), April 23-25, Spoleto, Italy, pp 471-472

Santangelo E, Fonzo V, Astolfi S, Zuchi S, Caccia R, Mosconi P, Mazzucato A, Soressi GP (2003a) The Cf-2 / Rcr3esc gene interaction in tomato (Lycopersicon esculentum) induces autonecrosis and triggers biochemical markers of oxidative burst at cellular level. Funct Plant Biol 30: 1117-1125


Santangelo E, Habashy AA, Caccia R, Mosconi P, Picarella ME, Mazzucato A, Soressi GP (2003b) Triploidi partenocarpici di pomodoro: una innovazione nel futuro del vivaismo orticolo. Italus Hortus 10: 191-194;

Santangelo E, Mazzucato A, Picarella ME, Mosconi P, Lioi L, Soressi GP (2006). Caratterizzazione del ‘Fagiolo del Purgatorio’ di Gradoli (VT). Italus Hortus 13: 496-502

Santangelo E, Antonelli A, Picarella ME, Soressi GP (2007a) The Lesion Mimic Mutants as a tool for unveiling the genic network operating during biotic and abiotic plant stresses. In: Thangadurai D, Tang W, Song SQ (eds) Plant Stress and Biotechnology (ISBN 978-81-89473-10-5). Oxford Book Company, Jaipur, India, pp 155-176

Santangelo E, Marabottini R, Antonelli M, Badiani M, Pasqualini S, Soressi GP (2007b) The autonecrosis triggered by Cf-2/Rcr3esc interaction in tomato (Solanum lycopersicum L.) involves an ethylene burst and the enzymatic machinery protecting the respiratory apparatus of mitochondria. In: Ramina A et al. (eds) Advances in Plant Ethylene Research: Proceedings of 7th International Symposium on the Plant Hormone Ethylene (Pisa, 18-22 June 2006): 391-392

Schettino M, Caccia R, Testa G, Soressi GP (1998) Piante di pomodoro transgeniche per l'inibitore di protease Kunitz, finalizzate al controllo dell'Helicoverpa armigera. Atti IV Giornate Scientifiche SOI, Sanremo, Imperia,1–3 Aprile, pp 505-506

Schettino M (2004) Trasformazione genetica del pomodoro con il gene inibitore di proteinasi Kunitz (KTI3) e suo effetto su sviluppo e sulla capacità riproduttiva di Helicoverpa armigera (Lepidoptera noctuidae) Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutors Proff. GP Soressi and C Pucci

Schiavi M (1977). Eredità di alcuni mutanti che determinano assenza di germogli laterali nella pianta di pomodoro (L. esculentum Mill.). Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Schiavi M, Soressi GP, Maggiore T, Casarotti D, Porcini F, Smedile E (1983) Utilizzazione del calore residuo da centrali termoelettriche per produzioni vegetali agrarie: risultati delle esperienze condotte a Tavazzano. Atti 1a Conferenza Internazionale Energia e Agricoltura, Milano 27-28-29 Aprile 1983 vol. 3-28: 1-25

Schiavi M, Rotino F, Parrini F, Casarotti D, Falavigna A, Maggiore T, Smedile E, Soressi GP (1984a) Use of waste heat from an electric generating plant (wet tower cooling) in agricolture: first results with Gerbera cultivation in glasshouse. Working Group “waste heat utilization” of E.S.N.A. Proceeding of the meeting held Piacenza 3-7 September 1984: 39-54

Schiavi M, Rotino GL, Parrini F, Casarotti D, Falavigna A, Maggiore T, Smedile E, Soressi GP (1984b) Use of waste heat from an electric generatine plant (open circuit cooling) in agricolture: two years of experiments with inbreds of maize. Proceedings E.S.N.A. working group “waste heat utilization” Piacenza 3-7 September 1984: 55-67

Schiavi M, Casarotti D, Maggiore T, Rotino GL, Soressi GP (1984c) Utilizzazione del calore delle acque residue della centrale termoelettrica di Tavazzano: un triennio di prove con fragola. Colture Protette 7: 31-38

Schiavi M, Masera R, Fantino MG, Lorenzoni C, Soressi GP (1984d) Metodi di selezione per tolleranza a Fusarium e per caratteristiche merceologiche del bulbo in cipolla. Atti Convegno GOF-SOI, 9 December 1983 Ferrara, pp. 27-30


Schiavi M, Masera R, Nervo G, Mazzola P, Soressi GP (1984e) Different efficiency of blow-flies and honey bees in controlled pollination of onion (Allium cepa L.). Eucarpia 3rd Allium Symposium, Wageningen, The Netherlands 4-6 Sept 1984, p 115

Schiavi M, Masera R, Lorenzoni C, Soressi GP (1988a) Metodi utilizzati per il miglioramento genetico della Dorata di Parma. Agricoltura e Ricerca 82: 21-30

Schiavi M, Masera R, Nerva G, Mazzola P, Soressi GP (1988b) Diversa efficacia pronuba dell’ape e della mosca cartaria in condizioni di impollinazione controllata della cipolla. Agricoltura e Ricerca 82: 35-38

Schiavi M, Vicini E, Tisselli V (1991) Miglioramento genetico: nuove varietà di cipolla. Agricoltura 19: 64-66 (Regione Emilia Romagna)

Schiavi M, Lorenzoni C, Soressi GP (1998) Onion. In: Italian Contribution to Plant Genetics and Breeding, GT Scarascia Mugnozza & MA Pagnotta (eds), pp 477-480. University of Tuscia, Viterbo

Scolari G (1987). Possibilità e limiti del genotipo “monostelo” nel miglioramento genetico del pomodoro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Sgherri C, Navari-Izzo F, Pardossi A, Soressi GP, Rizzo R (2007) The influence of diluted sea water and ripening stage on the content of antioxidants in fruits of different tomato genotypes. J Agric Food Chem 55: 2452-58

Somerville C, Browse J (1991) Plant lipids: metabolism, mutants, and membranes. Science 252: 80–87

Soressi GP (1964) Effetti genetici dei raggi X su alcune cultivar di pomodoro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. A Bianchi

Soressi GP (1966) Eterosi nel pomodoro e possibilità di ottenere ibridi senza demasculazione utilizzando marcatori genetici. Sementi Elette 12: 2-12

Soressi GP, Cravedi P, Marchesi G (1966) Descrizione di mutanti artificialmente indotti in cultivar di pomodoro. Genetica Agraria 20: 204-219

Soressi GP (1967a) Brown seed (bs) a tomato seed character which behaves as an endosperm trait. Tomato Genet Coop Rep 17: 50

Soressi GP (1967b) Spongy seed (ss), a new plant character which modifies the seed hair appearance. Tomato Genet Coop Rep 17: 49-50

Soressi GP, Cravedi P (1967) Tomato mutants obtained by means of X-ray and ethylmethanesulphonate (EMS) treatments. Tomato Genet Coop Rep 17: 51

Soressi GP (1968) Development of hybrid tomato seed making simultaneous use of the genetic markers brown seed and spongy seed. Tomato Genet Coop Rep 18: 37-38

Soressi GP (1969a) Il pomodoro. Trattato di genetica agraria speciale. Ed agricole, Bologna, 47 pp

Soressi GP (1969b) Utilità di alcuni caratteri mutanti nella costituzione di cultivar e di ibridi per la raccolta meccanica della coltura del pomodoro. Sementi Elette 15: 76-81

Soressi GP (1969c) I problemi e le prospettive della meccanizzazione della coltura del pomodoro con particolare riguardo alle operazioni di raccolta. Atti Gionate della Meccanizzazione della coltura del pomodoro, Camera di Comm di Piacenza, pp 5-15

Soressi GP (1970a) Tomato mutants following EMS seed treatments. Tomato Genet Coop Rep 20: 59


Soressi GP (1970b) Tomato hybrid seed production: a further scheme. Tomato Genet Coop Rep 20: 58-59

Soressi GP, Lorenzoni C (1970) Crossing rate after natural and hand pollination with and without emasculation. Tomato Genet Coop Rep 20: 61-63

Soressi GP, Borghi B, Lorenzoni C, Maggiore T, Mariani BM, Salamini F (1973) Ibridi di pomodoro determinati. II. Valutazione dell’attitudine combinatoria. Sementi Elette 19: 3-35

Soressi GP (1974) Pomodoro. In: Aspetti genetici dell’attività sementiera. Monografie di Genetica Agraria. Bianchi A, Pavia, II: 33-53

Soressi GP, Lorenzoni C, Borghi B (1974a) Valutazione di varietà e ibridi di anguria (Citrullus vulgaris L. Schrad) quale premessa al miglioramento genetico della specie. Sementi Elette 20: 9-19

Soressi GP, Gentinetta E, Odoardi M, Salamini F (1974b) Leaf peroxidase activities in tomato mutants affecting plant morphology. Biochem Genet 12: 181-198

Soressi GP (1975a) New spontaneous or chemically induced fruit ripening tomato mutants. Tomato Genet Coop Report 25: 21-22

Soressi GP (1975b) Un nuovo ideotipo di pomodoro per la raccolta meccanica. Atti Giornata orticola sul pomodoro da industria, Grosseto, Settembre 1974. Annali ISPORT, 6, 1-7

Soressi GP, Salamini F (1975) A monomendelian gene inducing parthenocarpic fruits. Rep Tom Genet Coop 25: 22

Soressi GP, Palmieri S, Falavigna A (1975) Una nuova tecnica per la produzione di seme ibrido di pomodoro. Atti della Giornata delle sementi orticole, Ascoli Piceno, 15-18 maggio 1975

Soressi GP (1976) Miglioramento genetico del pomodoro da industria. Conoscere per produrre 3: 7-12

Soressi GP (1977) Utilizzazione dei marcatori genetici. Riv di Agronomia 11: 52-57 Soressi (1979) Potential of mutation breeding in tomato. Israel-Italian joint meeting on

genetics and breeding of crop-plants. Monografia di Genetica Agraria, IV: 245-258 Soressi GP, Falavigna A (1979) Il miglioramento genetico dell’asparago (Precoce

d’Argenteuil). Atti giornate studio Coltura Asparago, Santena (TO), 13 Maggio 1978, pp. 7-11

Soressi GP, Schiavi M, Tosi A, Marchesi E (1981) Utilizzazione in orticoltura di acque calde reflue da centrali termoelettriche. Colture Protette 10: 15-19

Soressi GP, Rotino G, Schiavi M, Casarotti D (1985) Utilizzazione delle energie alternative: le acque reflue tiepide. Italia Agricola 122: 164-171

Soressi GP (1986a) Induced mutations and breeding in vegetable crops. Proc Hungarian-Italian Plant Genet Conf, June 26-30, pp 17-32

Soressi GP (1986b) Obiettivi e linee di sviluppo delle ricerche sull’asparago. Agricoltura Ricerca 8: 7-12

Soressi GP, Bruzzone GB, Frangi P (1986) Influenza della densità di coltivazione sulle caratteristiche morfologiche e produttive di genotipi di pomodoro monostelo e normale. Annali Facoltà di Agraria, Università Cattolica S. Cuore, Milano 26(1), 29-41.

Soressi GP (1988a) Progetto finalizzato MAF Orticoltura: sub-progetto Asparago e Cipolla. Agricoltura e Ricerca 10(92):43-60


Soressi GP (1988b) Progetto finalizzato orticoltura: sub progetto asparago e cipolla. Agricoltura e Ricerca 92: 43-60. Relazione presentata al convegno nazionale "Progetto finalizzato Orticoltura" Paestum (SA), 10-11 Novembre 1988

Soressi GP, Scarascia Mugnozza GT (1988) Energy saving through new plant architecture. Acta Hortic 229: 39-50

Soressi GP, Pardossi A, Portesi A (1988) "Monostem" tomato: a plant model for the high technology Era. International Symposium on High Tecnhology in Protected Cultivation, Hamamatsu, Japan, May 12-15, 1988 (Book of abstract 2-3)

Soressi GP, Porcelli S (1988) Obiettivi e linee di sviluppo delle ricerche sulla cipolla. Agricoltura e Ricerca 82: 1-4

Soressi GP, Schiavi M, Falavigna A (1991) Le colture in vitro e la micropropagazione per il rinnovamento del vivaismo e della ortoflorofrutticoltura. Terra e Vita, Speciale micropropagazione, 1: 78-82

Soressi GP (1992) Origine, evoluzione, importanza del pomodoro coltivato. pp. 129-146. In “Atti Convegno Internazionale "Scambi Floristici fra Vecchio e Nuovo Mondo. Riflessi Agro-Selvicolturali e impatti naturalistico-ambientali e paesaggistici". Genova, 22-23 Aprile 1991. Fondazione Reg. Cristoforo Colombo, Comune - Ente fiera Genova

Soressi GP, Mapelli S (1992). Genetical, physiological and biochemical aspects of a monostem tomato phenotype suitable for mechanization and processing. Acta Horticulturae, 301, 229-236

Soressi GP, Campolmi C, Pardossi A, Tognoni F (1992) Nuovi modelli di piante per le moderne tecniche colturali. Colture Protette 21 (3): 51-57

Soressi GP (2003) Variabilità disponibile e utilizzabile nel miglioramento genetico del carciofo. L’Informatore Agrario 22: 47-50

Soressi GP, Papalini P, Piccioni C, Santangelo E, Mosconi P, Picarella EM, Mazzucato A (2007) Innovazione varietale e programmazione colturale. In atti del convegno “Ricerca ed innovazione per la valorizzazione del carciofo nella regione Lazio”, ARSIAL Tarquinia 30 novembre 2007: 12-23

Soressi GP, Cammareri G, Picarella MP (2008) Improvement of in vitro vegetative propagation technique in tomato (Solanum lycopersicum). Acta Horticulturae (in press)

Sozzi A (1969) Impiego del carattere seme bruno (bs) nella produzione di seme ibrido di pomodoro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Sozzi A, Gorini FL, Soressi GP (1981) Performance of nor2 hybrids of tomato (Lycopersicon esculentum Mill.) during storage at controlled temperature. Acta Hort 116: 223-232

Speranza S (2001) Dinamica di popolazione della Nottua gialla del pomodoro (Helicoverpa armigera (Hüb.) Lepidoptera, Noctuidae) nel Centro Italia mediante applicazione di Traptest® innescate con due diverse miscele feromoniche. Informatore Agrario 22/2001: 59-62

Stamegna L (2008) Analisi genetiche in un mutante maschiosterile condizionale di pomodoro (Solanum lycopersicum L.). Graduation Thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. A Mazzucato

Stancanelli G (1989) Analisi di crescita a livello di plantula in asparago. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi


Stancanelli G, Falavigna A, Soressi GP (1990) Growth analysis of seedlings and spears in different asparagus genotypes. Acta Horticulturae 271: 503-508

Testa G (1998). Ricerca di geni differenzialmente espressi in mutanti partenocarpici di pomodoro a confronto con linee quasi isogeniche normali. Specialization thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Testa G, Caccia R, Tilesi F, Soressi GP, Mazzucato A (2002) Sequencing and characterization of tomato genes putatively involved in fruit set and early development. Sex Plant Reprod 14: 269-277

Tiburtini S (1992) Variabilità somaclonale indotta nella cv. Alice di pomodoro a seguito di rigenerazione in vitro di espianti cotiledonari. Graduation thesis, Università della Tuscia, Viterbo, Italy. Tutor Prof. GP Soressi

Todisco F (1978) Eredità di alcuni mutanti “sundwarf” e loro utilizzazione nel miglioramento genetico del pomodoro. Graduation thesis. Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Venezia A (1991) Raccolta e classificazione botanica di germoplasma di asparago spontaneo in Sicilia: messa a punto delle tecniche di germinazione e moltiplicazione in vitro. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Venezia A, Soressi GP, Falavigna A (1993) Aspetti relativi alla valorizzazione di specie di asparago spontanee in Italia. Agricoltura Ricerca 141: 41-48

Ziliotti A (1984) Interazione tra mutanti di pomodoro che controllano l’emissione di germogli laterali (ls e to-2) e la formazione partenocarpica della bacca (pat e pat-2). Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi


Memorandum The graduation theses tutored by Prof. GP Soressi not included in the contributions are

here reported: Francavilla S (1992) Rigenerazione in vitro e trasferimento di geni marcatori via

Agrobacterium tumefaciens in Solanum spp. per il miglioramento genetico della melanzana. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Ferrari F (1996) Le nuove piante da fronda: problematiche inerenti la moltiplicazione vegetativa con particolare riferimento al Pittosporum tenuifolium cv. silver queen. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Parodi A (1992) Miglioramento genetico dell’Arginanthemum frutescens attraverso mutagenesi. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutor Prof. GP Soressi

Pecchioni N (1980) Analisi della sintesi proteica in condizioni di stress termici in Gerbera jamesonii hybrida H. Bolus. Graduation thesis, Università Cattolica Sacro Cuore, Piacenza, Italy. Tutors Proff. GP Soressi and N Marmiroli

Santangelo E (1994) Rivalutazione e valorizzazione del cece attraverso il ricorso a individuazione di un protocollo di trasformazione in vitro per consentire l’introduzione di geni utili. PhD thesis, Università della Tuscia, Viterbo, Italy. Tutors Proff. GP Soressi and E Porceddu


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