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Currently Apulia region accounts about 59 winegrape cultivars authorised for growing, including historical varieties. The existence of autochthonous minor varieties in the region has been recognized and is recently at the centre of a program aiming to their recovery and their further exploitation for commercial use. In the present study, 87 traditional, autochthonous and minor Apulian grapevine accessions were examined for their genetic profiles. Among them, 64 unique genotypes were found. Synonyms among Apulian genotypes and Italian varieties cultivated in Apulia or other Italian regions were identified by searching for genetic matching in different molecular database publicly available and through bibliographic search with published genetic data. The main goal of the current study was to identify, characterise and catalogue local germplasm rich of economical and historical significance.

The ambitious idea for the development of new tablegrape seedless varieties, suitable for growing in the Mediterranean basin and highly competitive on the international markets, has recently led to the foundation (in 2015) of the Italian Variety Club (IVC) a network of 17 enterprises and 2 research Institutions in Southern Italy. This synergic collaboration among private companies and researchers aims to develop and transfer varietal and technical innovation to the growers and to strengthen the Italian table grape supply chain merging high quality products and sustainable agriculture. The breeding program is organized in different interconnected activities such as: "seed x seedless" varieties and "seedless x seedless" varieties controlled crosses, involving Apulian traditional genotypes and élite cultivars; F1seedling production with molecular marker assisted selection (MAS) for an early identification of seedless genotypes; in vitro embryo culture; plant evaluation under field conditions. In the coming years the activities of varietal description and official registration, Patent protection, and establishment of stock fields will be initiated. As result of the 3-year activities, the progenies of 75 "seed x seedless" varieties crosses, 28 "seedless x seedless" varieties crosses and 12 crosses involving germplasm with resistance traits have been obtained and analysed and the field evaluation begun. With regard to progenies derived from "seedless x seedless" varieties crosses, the average embryogenesis percentages (no. of germinated embryos/no. of cultivated ovules) was about 8-9%, varying according to the parents used. Following the extensive use of MAS about 40% of each "seed x seedless" varieties progeny carried allele associated to seedless locus. Actually about 6,000 new genotypes are grown in the first selection field, while further 6,000 seedless plants are going to be planted within 2018.

In recent years, an increasing number of typicality marks has been awarded to high-quality olive oils produced from local cultivars. In this case, quality control requires effective varietal checks of the starting materials. Moreover, accurate cultivar identification is essential in vegetative-propagated plants distributed by nurseries and is a pre-requisite to register new cultivars. Food genomics provides many tools for cultivar identification and traceability from tree to oil and table olives. The results of the application of different classes of DNA markers to olive with the purpose of checking cultivar identity and variability of plant material are extensively discussed in this review, with special regard to repeatability issues and polymorphism degree. The characterization of olive germplasm from all countries of the Mediterranean basin and from less studied geographical areas is described and innovative high-throughput molecular tools to manage reference collections are reviewed. Then the transferability of DNA markers to processed products – virgin olive oils and table olives – is overviewed to point out strengths and weaknesses, with special regard to (i) the influence of processing steps and storage time on the quantity and quality of residual DNA, (ii) recent advances to overcome the bottleneck of DNA extraction from processed products, (iii) factors affecting whole comparability of DNA profiles between fresh plant materials and end-products, (iv) drawbacks in the analysis of multi-cultivar versus single-cultivar end-products and (v) the potential of quantitative polymerase chain reaction (PCR)-based techniques.

The accurate description of plant biodiversity is of utmost importance to efficiently address efforts in conservation genetics and breeding. Herein, we report the successful application of a genotyping-by-sequencing (GBS) approach in chickpea (Cicer arietinum L.), resulting in the characterization of a cultivated germplasm collection with 3,187 high-quality single nucleotide polymorphism (SNP) markers. Genetic structure inference, principal component analysis and hierarchical clustering all together indicated the identification of a genetic cluster corresponding to black-seeded genotypes traditionally cultivated in Southern Italy. Remarkably, this cluster was clearly distinct at both genetic and phenotypic levels from germplasm groups reflecting the commercial chickpea classification in desi and kabuli seed types. Fixation index estimates for individual polymorphisms pointed out loci and genomic regions that might be of significance for the diversification of agronomic and commercial traits. Overall, our findings provide information on genetic relationships within cultivated chickpea and highlight a gene pool of great interest for the scientific community and chickpea breeding, which is limited by the low genetic diversity available in the primary gene pool

In questo lavoro è stata condotta la caratterizzazione genetico-molecolare di circa 100 accessioni di olivo campionate in diverse località del Salento. L'obiettivo è stato la caratterizzazione, mediante marcatori molecolari (microsatelliti), di varietà autoctone del Salento a rischio di erosione genetica, e di altre varietà di olivo che, seppure di diversa origine geografica, risultano coltivate nel territorio salentino.

Abscisic acid (ABA) is associated with regulating plant adaptive responses to various environmental stresses. In particular, drought stress signals are transmitted through at least two pathways: one is abscisic acid (ABA)-dependent, and the other is ABA-independent. In the first case, drought stress increases the cellular ABA levels, which induces the expression of drought stress-responsive genes, such as 9-cis-epoxycarotenoid dioxygenase (NCED) and zeaxanthin epoxidase (ZEP). These genes belong to the carotenoid biosynthesis scenario. To date, most research of grapevines has focused on the physiological mechanisms of ABA during fruit ripening. Our interest is on studying the role of NCED and ZEP genes as candidate genes exhibiting up-regulation upon drought-stressed conditions. At the same time, several plant physiological parameters, such as leaf water status (Ψ1), net assimilation rate (A), stomatal conductance (gs), transpiration rate (E), and soil water potential (Ψs), were monitored. To explain the complex molecular pattern undergoing these physiological changes, we investigated the levels of expression of one candidate gene encoding for VvNCED1. The results provided evidence of a different transcriptional pattern of the gene between the control and stressed plants, leading to a major accumulation of NCED1 transcripts in the stressed plants.

In questo lavoro ci siamo occupati del recupero del germoplasma olivicolo salentino ancora presente in provincia di Lecce, ma a forte rischio di erosione genetica. In particolare, abbiamo studiato il profilo molecolare di varietà antiche, quali: Colozzese, Barone di Monteprofico, Ornella, Usciana, Cornola, Uliva grossa, e di diversi cloni delle cultivar autoctone più diffuse sul territorio, ovvero Cellina di Nardò e Ogliarola di Lecce. Tutte le varietà analizzate in questo studio sono conservate in un campo collezione allestito negli anni '90 in agro di Carpignano salentino.

Nell"ambito del Piano di sviluppo rurale 2007-2013, la regione Puglia ha identificato nella misura 214/4 sub-azione a): "Progetti integrati per la biodiversità" l"importanza di tutelare, caratterizzare e identificare il numeroso germoplasma autoctono regionale. La regione presenta un territorio molto vasto e ricco di germoplasma autoctono vegetale, che corre il rischio di essere soggetto ad una graduale erosione genetica e quindi ad una perdita di biodiversità irreversibile. L"obiettivo principale dei progetti Re.Ge.Vi.P., Re.Ger.O.P., e Re.Ge.Fru.P., occupandosi rispettivamente di vite, olivo e fruttiferi, è di realizzare, attraverso un approccio integrato multidisciplinare, un effettivo recupero e studio del germoplasma propedeutico alla valorizzazione della agrobiodiversità regionale. La prima fase del lavoro consiste in una minuziosa ricerca bibliografica in fonti edite e inedite, ma anche attraverso le testimonianze e la memoria storica degli agricoltori su usi e tradizioni concernenti le vecchie varietà coltivate. La ricerca, l"individuazione sul territorio e georeferenziazione del germoplasma stanno consentendo di recuperare il materiale arricchendo le collezioni ex situ già esistenti. È inoltre previsto che tutto il materiale recuperato sia caratterizzato mediante approcci morfologici, molecolari, agronomici tecnologici e sanitari al fine di verificare l"identità varietale, chiarire l"esistenza di omonimie e sinonimie, scoprire nuova biodiversità intra-varietale, registrare ufficialmente le nuove accessioni di germoplasma nonché valutarne le potenzialità economico-produttive per una possibile ripresa in coltivazione. Nell"ambito dei progetti, oltre a costituire delle collezioni ex situ di riferimento per l"intera regione, secondo le linee guida rilasciate nel 2012 dal Gruppo di lavoro per le "Risorse genetiche vegetali" del MiPAAF, l"inserimento del germoplasma in collezioni private e presso aziende custodi consentirà una conservazione bilanciata tra attività in situ ed ex situ. Infine tutte le nuove informazioni acquisite, catalogate in un archivio elettronico fruibile al pubblico, costituiranno al contempo un dettagliato strumento di conoscenza e divulgazione dell"enorme patrimonio viticolo, olivicolo e frutticolo regionale. Nel prossimo programma di sviluppo rurale pugliese, si auspica che i risultati dei progetti integrati, insieme alla recente legge regionale di "Tutela delle risorse genetiche autoctone" (L.R. n.39 dell"11/12/2013), possano concretizzarsi in reali iniziative di valorizzazione del germoplasma locale.

Italian law (1) prohibits the manufacture of pasta containing more than 3% soft wheat (Trilicum aestivum L) for the domestic market, while it allows import-export of pasta totally or partially prepared from soft wheat, in this case requiring to clearly indicate it in the label. Consequently, a strong interest towards the detection of soft wheat has stimulated the development of numerous analytical methods, generally aimed at searching specific protein fractions, the first set up as early as at the end of 1960s (2). More recently, a new generation of methods which employ DNA screening for sequences localized in the D-genome. characteristic for common wheat, has become available. Microsatellites are sequences of repeated DNA that can be analyzed by means of a single PCR reaction, providing short-sized arnplicons (3). The small size of generated amplicons (around 200 base pairs) is crucial if analyzing food samples submitted, during their production process, to high temperature and/ or strong mechanical treatments, such in case of pasta. The arm of this work was to evaluate the possibility to apply the analysis of DNA microsateUites to set up a method of detection of soft wheat in semolina and pasta. The first step consisted in DNA extraction from flour and semolina from various cultivars of soft and durum wheat (Triticum lurgidum L. var. durum), and from commercial pasta, by comparing CTAB laboratory protocol and the commercial kits NucleoSpin Plant (Macherey-Nagel), NucleoSpm Food (Macherey-Nagel) and Gene Elute Plant (Sigma). Their protocols have been modified to better adapt to the food sample considered. The better efficiency, in terms of extraction yield and DNA quality and purity, was obtained using NucleoSpin Food, which gave an average yield of 275 ug DNA/g sample. Then, nine primer pairs amplifying microsatellite sequences, chosen for being localized on D-genome according to literature data, were tested on the DNA extracted, with the aim of verifying their effectiveness in distinguishirsgsoft wheat from durum one. The obtained results enabled to select an efficient D-genome-specific microsatellite sequence able to detect common wheat. A polymorphism of the type "presence/absence" was observed, with no amplification in durum wheat (negative control), and an amplicon of 210 base pairs present solely in samples deriving from soft wheat. The first trials were carried out by qualitative PCR, able just to assess presence or absence of soft wheat with a threshold of 3%, but not adequately able to quantify it. Subsequently, semolina and pasta samples were prepared from blends of durum wheat:soft wheat in the ratios 60:40, 70:30, 80:20, 90:10, 95:5, 97:3, and 97.5:2.5, and SYBR Green real-time PCR was used to quantify the soft wheat adulteration. In this way the threshold was lowered to 2.5%. We are currently proceeding to a further improvement of the method by a TaqMan approach.

Italian industrial pasta has to be prepared using exclusively durum wheat (Triticum turgidum L. var. durum) semolina. The Italian law only allows the presence of maximum 3% soft wheat (Triticum aestivum L.) as an accidental contamination during the storage of raw material. Traditionally, the presence of soft wheat contamination is assessed through enzymatic methods, which are fairly cumbersome and do not allow an accurate quantification of the contaminant. Recently, a new generation of methods which employ DNA screening for sequences localized in the D-genome, characteristic for soft wheat, has become available, including real-time quantitative PCR analyses (Alorio et al. 2003; Terzi et al. 2003). In a previous study, a microsatellite sequence specific of the D genome had been chosen, among others, for traceability of soft wheat in qualitative analysis and quantitative Sybr green based real-time experiments (Pasqualone et al. 2007). However, a good quantification was only obtained for semolina samples as compared to processed food. In this work, we describe an improved method based on the same soft wheat genomic region, using an alternative approach. The DNA region was cloned and sequenced, and primers and a dual-labeled (Taqman(TM)) probe were designed nearby the microsatellite repeat. Semolina and pasta samples were prepared from several blends of durum wheat:soft wheat in controlled ratios, and genomic DNA extraction was optimized to recover a high DNA amount. The Taqman(TM) approach allowed to avoid aspecific amplifications due to primer dimer formation, and therefore produced a more accurate estimate of soft wheat contamination in semolina based products.

The detection of common wheat (Triticum aestivum L.) contaminations in durum wheat (Triticum turgidum L. var. durum) semolina has always been the object of interest and stimulated the development of numerous analytical methods, generally by searching specific common wheat protein fractions. These methods were mainly aimed to preserve pasta "purity" and check its adherence to Italian rules. Recently, the typical Altamura bread (Apulia, Italy), has been awarded with the European Protected Designation of Origin (PDO) mark, owing to its typicality. This product should be prepared using only durum wheat semolina, and should not contain soft wheat flour. The aim of this work was to evaluate the possibility to apply DNA microsatellite analysis to set up a method for the detection of soft wheat in durum wheat semolina. This strategy can be applied for checking the raw materials used for Altamura bread, as well as for Italian pasta preparation. A total number of 9 primer pairs amplifying microsatellite sequences was chosen for being localized on D-genome according to literature data, and was tested on the DNA extracted from semolina and flour of various durum wheat and soft wheat cultivars, with the aim of verifying the effectiveness in distinguishing common wheat from durum one. The obtained results allowed to select, among them, an efficient D-genome-specific microsatellite. SYBR Green real-time PCR enabled to successfully detect common wheat in semolina, with a threshold of 2.5%.