In 1967, following the law N. 1318 published in G. U. n. 14 dated 18 January 1968, the Horticultural Experimental station of Milano University until then belonging to the Italian Ministry of Agriculture, was replaced by the Montanaso Lombardo Section of the Research Institute of Vegetable crops. After the application of the law n. 454/1999, the aforesaid Section was joined to “Agricultural Research Counsil (CRA) with the name “CRA – Reserch Unit for Vegetable Crops (CRA-ORL).
CRA-ORL d disposes of a farm (about 17 ha), two sheds (800 m2), four glasshouses (1,800 m2), two labs (200 m2) equipped for in vitro tissue culture activity (four growth chambers, seven positions at laminar flow cabinets; microscope, cytofluorimeter). In the lab there are also: one automatic dispenser of liquids to prepare the reactions, thermocyclers for qRT-PCR e HRM-PCR reactions. Recently an AdvanCE™ FS96 devices allowing an high output automatic quality and quantity analysis of nucleic acids (genomic DNA, total RNA and DNA fragments).
In 2011 the persons with permanent position were: one director incharge, three researchers, three technicians, three tecnical operators, one administrative operator. There were also: four persons with formation contract (felloship, research contract), two Ph. D. student and five seasonal workers.
CRA-ORL carries out vegetable and industrial crops breeding, following conventional and advanced methods; studies bichemical, technological and agronomical methods aimed to lower the environmental impact of crop cultivation with particular emphasis to asparagus, onion, bean, eggplant and pepper for new cultivars development and for genetic and merchandise characterization of local cultivars. CRA-ORL collaborates with the CRA – Genomic and Post –genomic Research Center of Fiorenzuola d’Arda to develop new molecular biologicak tools to be applied to vegetable crop breedind.
The main research activity developed and still in progress at CRA-ORL, concerned/s vegetable crop breeding, which have been performed with the support of different biotechnological methods (in vitro tissue culture and molecular genetics).
Asparagus (Asparagus officinalis). The breeding program began in 1974 and is based on the following steps: production of doubled-haploid (DH) clones following in vitro anther culture; synthesis of F1 all-male hybrids by crossing DH clones; micropropagation of F1 hybrid DH parents for commercial seed production. A collection of 200 DH clones, selected for morphological traits and resistance to diseases, have been selected and utilized as parents of F1 all-male hybrids. At present the best three CRA-ORL hybrids (Eros, Franco, Ercole) represent near the whole asparagus growing area of North Italy; moreover the all-male hybrid Italo in rapidly spread in South Italy. The in vitro anther culture of asparagus has also successful applied for reducing to diploid the tetraploid level of plants derived from interspecific hybridization with A. maritimus and A. acutifolius. Di-haploid clones so obtained have been successfully crossed with DH ones, with the aim to introgress useful traits from the two relatives into the cultivated species. Since 2006 a genetic characterization on the whole CRA-ORL A. officinalis genotypes collection with AFLP molecular markers and a phylogenetic study on wild materials using SNPs markers were performed. Moreover two cDNA libraries were developed, through the use of recent sequencing strategies, from which molecular markers (SNPs and SSRs) putatively associated to agronomic interesting traits (sex determination and Puccinia asparagi resistance) were isolated.
Onion (Allium cepa). During 1975-1990 period the breeding activity of the Italian local varieties “Dorata di Parma” and “Borettana” allowed to release commercially five varieties, out of which “Marica”, “Morgana” and “Sorriso” are still widely spread both in Italy and abroad. Recently two new varieties (Monica and Morena) from the local ecotype “Rossa di Tropea” and some synthetic populations from the local varieties “Rossa di Breme” and “Paglierina di Sermide” were selected for morphological traits. At CRA-ORL the production of doubled haploid lines from in vitro ovule culture was set up. At present a number of DH gynogenic lines are going to be utilized as parent of F1 hybrids in collaboration with a seed company.
Common Bean (P. vulgaris). From 1970 to 1996, breeding activity aimed at the introgression of genetic traits conferring resistance to Bean Common Mosaic Virus (BCMV). Four borlotto- and three canellino-type determinate varieties were released. Two of them, Giulia (borlotto-type) and Montalbano (canellino-type), still maintain commercial interest. Several climbing lines resistant to BCMV and determinate lines resistant to BCMNV (necrotic) strains were also selected. The breeding work done during the last fiftheen years was mainly focused at the nutritional improvement of bean seed as well as the valorization of Italian landraces. Concerning the first objective, the genetic traits “lectin absence”, “low tannin” and “low phytate” seed content, have been introgressed into high yielding lines, previously selected from CIAT accessions. The breeding lines so far obtained are going to be studied by agro-industries for food and feed purposes. The valorization of ten Italian landraces was pursued by performing the following activities: conservative selection, eradication of seed-transmitted pathogens through the elimination of infected plants, analysis of culinary technological parameters, morphological and molecular genetic characterization of the germplasm collections.
Eggplant (Solanum melongena). The breeding activity started in 1985. The conventional methods based on intraspecifc crosses, artificial pathogen infection and selection, were successfully integrated with: - in vitro culture of anthers to obtain doubled haploid plants from segregating lines; - protoplast fusion to overcome the sexual incompatibility between S. melongena S. aethiopicum and S. integrifolium - in vitro culture of anthers for obtaining di-haploid lines from somatic hybrids obtained through protoplast fusion; - PCR-based analyses for both structural and functional genomic studies, - expression analysis of genes of interest involved in the plant-pathogen interaction and in other metabolic pathways; - genetic engineering for the introduction in the eggplant genome of genes of interest. The following genetic materials were at present available: - trasgenic parthenocarpic lines for the presence of DefH9-iaam gene (international patent); - transgenic male-sterile lines through miRNA-mediated silencing of TAF10 and TAF13 basal transcription factors (international patent); - androgenetic breeding lines derived from in vitro anther culture; - di-haploid lines regenerated from in vitro anther culture of tetraploid interspecific hybrids; - lines resistant to Fusarium oxysporum f.sp. melongenae from advanced crosses between di-haploid and recurrent diploid breeding lines; - breeding lines tolerant to Verticillium dahliae from sexual crosses with S. sodomaeum and with Asiatic accessions. The selection of lines for agronomical traits as well as for resistance to soilborne pathogens are mainly carried out in the framework of joined projects with seed companies, with the aim of generating F1 hybrids. The first hybrid, ‘Sabelle’, is going to be released commercially. During the last five years molecular markers associated to resistance to Fusarium oxysporum f.sp. melongenae were developed and a panel group of eggplant genes involved in the interaction with Fusarium oxysporum f.sp. melongenae and Verticillium dahliae identified. Moreover, a molecular genetic linkage map based on AFLP, COS, SSR, CAPS e SNP markers was developed and QTLs linked to genes controlling agronomic traits by using both segregation populations and “association” approaches are under study.
Pepper (Capsicum annuum). The breeding programme was started in 1990 with the main purpose to introgress the genetic resistance to virus (TSWV, PVY, CMV), nematodes (Meloidogyne spp.) and fungi (Phytophthora capsici, Verticillium dahliae) into commercially important local Italian varieties: Corno di Carmagnola, Cuneo, Quadrato d’Asti. To this purpose intra- and inter- specific crosses and back-crosses to recurrent parents were carried out. The resistance level of lines were tested after artificial inoculation of pathogen; while agronomical performances of resistant plants were evaluated in the field. In vitro anther culture were extensively applied to stabilize segregating lines. The following genetic material is now available: Breeding lines with multiple resistant to virus, Phytophthora capsici and nematodes. Following selfing or anthers in vitro culture, homozygous lines of the local variety Friariello, Nocera, Senise, Roggianese, Topepo and Cornetto Calabrese, were obtained. Such lines were characterized for morphological and agronomical traits and genetic similarity following molecular markers.
Collaboration with Regions. During the last fifteen years CRA-ORL has mantained the scientific responsability of the projects funded by Lombardia and Piemonte Regions, aimed at the vegetable crops varietal trials; besides collaborated with Veneto Agricoltura e Centro di Ricerche Produzioni Vegetali (CRPV) with reference to similar activity carried out in Veneto and Emilia Romagna Regions.
Collateral activity carried out at CRA-ORL is mainly focused on the maintenance of Italian vegetable crop local varieties and accessions with potential interest for breeding.
Asparagus: - spontaneous species of the Mediterranean Basin; - parental clones of the hybrids released at CRA-ORL; doubled-haploid androgenetic (DH) clones derived from in vitro anther culture; interspecific hybrid obtained for the cross A. officinalis, A. maritimus, A. acutifolius and relative off-springs; di-haploid androgenetic (DI) clones obtained form in vitro anther culture of tetraploid interspecific plants; off-springs obtained by crossing DH and DI clones.
Onion: thirty local varieties from differ areas of Italy; - about fifty doubled-haploid gynogenetic lines, potentially asefull as parents of F1 hybrids; fiftheen synthetic populations and several S1 lines of the ecotypes “Rossa di Tropea”, “Paglierina di Sermide” and “Rossa di Breme”.
Bean: - a number of old landraces coming from different Italian areas; - varieties, developed by CRA-ORL since 1970; - two P. coccineus lines exhibiting determinate growth habit, one white seeded and the second colour seeded (black mottled on wine pink background); - breeding lines developed at CRA-ORL (resistant to BCMV and BCMNV, lectin-null and/or α-Amilase Inhibitor, low phytic acid (lpa), white or coloured seed)
Eggplant: - the following relatives: S. sodomeum, S. aetiopicum, s. integrifolium, S. indicum, S. viarum, S. macrocarpa, S. torvum; - about one-hundred accessions of S. melongena characterized by: tolerance to Verticillium dahliae and Fusarium solani, different shape and fruit color and plant habitus.
Pepper: - the following relatives: C. chinense, C. frutescens, C. baccatum, C. chacoense, C. praetermissum, C. pubescens; - accessions of C. annuum resistant to Phytophthora spp., nematodes and viruses; a number of Italian local varieties from different part of Italy.
Tomate: - ten tablr tomate local varieties with big fruit; - one-tundre-twenty accessions collected in Italy and abroad, characterized by different fruit shape, size, chemical coposition and color; different plant development and cultural cycle.
Melon (Cucumis melo var cantalupensis: - several near homogygous lines belongings to the following traditional tipologies: Viadana “Moscatello Mantovano” “Melone di Cavenzano “; “Banana di Piacenza”, “Zatta”, “Rampichino”, Americano Bianco and other less important; about twenty F1 hybrids derived from combination of Viadana lines.
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