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Grapevine (Vitis vinifera L.) is one of the oldest cultivated plants, with wide genetic diversification and phenotypic plasticity that remains largely under-explored. Since ancient times, Apulia has served as a central hub for commercial exchanges that have resulted in a rich heritage of grape genetic resources adapted to the pedoclimate conditions of the region. Information on the genetic variation and knowledge of the structure of V. vinifera populations is essential to manage and preserve gene pools and to recover alleles left behind by selective breeding. In this study, we applied the genotyping-by-sequencing (GBS) technique on a large grapevine collection including Apulian autochthonous cultivars and clones of some important local cultivars. Work is in progress to generate a high-quality single-nucleotide polymorphism (SNP) dataset that will be used to assess genetic diversity, determine population structure and estimate intraclonal variability. In addition, SNP markers will be combined with phenotypic data via genome-wide association studies in order to discover genetic loci associated with key agronomic traits.

Italian spumante has a long tradition, deeply rooted in the Piedmont region since the 19th century. A fundamental aspect of spumante production is berry acidity, since it influences both the quality and taste of spumante. The acidity of the berry results from the accumulation of H+ ions in the vacuole of the cell, and this process involves complex biochemical changes not yet completely understood. Furthermore, acidification is strongly influenced by environment; in particular, higher temperature is associated with lower berry acidity. As part of a project aimed at evaluating and promoting Apulian cultivars for spumante production, six autochthonous grapevine cultivars (‘Bianco d’Alessano’, ‘Bombino Nero’, ‘Maresco’, ‘Minutolo’, ‘Negramaro’ and ‘Uva di Troia’) and two international cultivars (‘Pinot Blanc’ and ‘Pinot Noir’) were characterized both genetically, to constitute molecular fingerprints, and biochemically, to detect berry acidity values. The results obtained allowed us to identify the autochthonous cultivar ‘Maresco’ as the best candidate for comparison with the most commonly used cultivar for spumante production at the international scale, ‘Pinot Blanc’.

Seedlessness represents a highly appreciated trait in table grapes. Based on an interesting case of seedless fruit production described in the crop speciesAnnona squamosa, we focused on theVitis vinifera INNER NO OUTER (INO)gene as a candidate. This gene encodes a transcription factor belonging to theYABBYfamily involved in the determination of abaxial identity in several organs. InArabidopsis thaliana, this gene was shown to be essential for the formation and asymmetric growth of the ovule outer integument and its mutation leads to a phenotypic defect of ovules and failure in seed formation. In this study, we identified in silico theV. viniferaorthologue and investigated its phylogenetic relationship toINOgenes from other species and its expression in different organs in seeded and seedless varieties. Applying cross-species complementation, we have tested its functionality in the Arabidopsisino-1 mutant. We show that theV. viniferaINO successfully rescues the ovule outer integument growth and seeds set and also partially complements the outer integument asymmetric growth in the Arabidopsis mutant, differently from orthologues from other species. These data demonstrate that VviINO retains similar activity and protein targets in grapevine as in Arabidopsis. Potential implications for grapevine breeding are discussed.

Cultivation of faba bean (Vicia faba L.) in Tunisia is largely based on improved varieties of the crop. However, a few farmers continue to produce local cultivars or landraces. The National Gene Bank of Tunisia (NGBT) recently launched a collection project for faba bean landraces, with special focus on the regions of the North West, traditionally devoted to cultivating grain legumes, and where around 80% of the total national faba bean cultivation area is located. The seed phenotypic features of the collected samples were studied, and the genetic diversity and population structure analyzed using simple sequence repeat markers. The genetic constitution of the present samples was compared to that of faba bean samples collected by teams of the International Center for Agricultural Research in the Dry Areas (ICARDA) in the 1970s in the same region, and stored at the ICARDA gene bank. The results of the diversity analysis demonstrate that the recently collected samples and those stored at ICARDA largely overlap, thus demonstrating that over the past 50 years, little genetic change has occurred to the local faba bean populations examined. These findings suggest that farmers serendipitously applied international best practices for in situ conservation of agricultural crops.

Information on the distribution of genetic variation is essential to preserve olive germplasm from erosion and to recover alleles lost through selective breeding. In addition, knowledge on population structure and genotype–phenotype associations is crucial to support modern olive breeding programs that must respond to new environmental conditions imposed by climate change and novel biotic/abiotic stressors. To further our understanding of genetic variation in the olive, we performed genotype-by-sequencing on a panel of 94 Italian olive cultivars. A reference-based and a reference-independent SNP calling pipeline generated 22,088 and 8,088 high-quality SNPs, respectively. Both datasets were used to model population structure via parametric and non parametric clustering. Although the two pipelines yielded a 3-fold difference in the number of SNPs, both described wide genetic variability among our study panel and allowed individuals to be grouped based on fruit weight and the geographical area of cultivation. Multidimensional scaling analysis on identity-by-state allele-sharing values as well as inference of population mixtures from genome-wide allele frequency data corroborated the clustering pattern we observed. These findings allowed us to formulate hypotheses about geographical relationships of Italian olive cultivars and to confirm known and uncover novel cases of synonymy.

The morphology of olive flowers allows either self- or cross-pollination that could partially explain the existence of both reproductive features in this species. However, a high degree of self-incompatibility is reported for many olive genotypes, that could be an important reproductive barrier influencing olive yield. Due to the strong environmental influence, results of compatibility tests are often contradictory, making cultivar classification quite imprecise. In this study, the self-incompatibility value has been determined for four olive genotypes (Bella di Spagna, Coratina, Leccino, and Ogliarola barese) widespread in the Mediterranean basin. Moreover, the incompatibility relationships of cultivar Coratina with some suitable pollinizers (Leccino, Oliastro, and Picholine) have been studied in controlled crosses: the in vitro germination potential of progenies has been evaluated and the selection of cross-derived embryos has been indirectly performed by the molecular characterization of the corresponding endosperm. The results increase knowledge on factors affecting self-compatibility in olive. Moreover, they provide useful information to farmers about the most effective cultivars for the set-up of new olive grove or for graft planning. Finally, they provide a new strategy and procedure based on endosperm analysis by SSRs for an accurate, fast, and relatively cheap screening of embryos/seedlings.

Cyclic AMP plays important roles in different physiological processes, including plant defence responses. However, as little information is known on plant enzymes responsible for cAMP production/degradation, studies of cAMP functions have relied, to date, on non-specific pharmacological approaches. We therefore developed a more reliable approach, producing transgenic Arabidopsis thaliana lines overexpressing the 'cAMP-sponge' (cAS), a genetic tool that specifically buffers cAMP levels. In response to an avirulent strain of Pseudomonas syringae pv. tomato (PstAvrB), cAS plants showed a higher bacterial growth and a reduced hypersensitive cell death in comparison with wild-type (WT) plants. The low cAMP availability after pathogen infection delayed cytosolic calcium elevation, as well as hydrogen peroxide increase and induction of redox systems. The proteomic analysis, performed 24 h post-infection, indicated that a core of 49 proteins was modulated in both genotypes, while 16 and 42 proteins were uniquely modulated in WT and cAS lines, respectively. The involvement of these proteins in the impairment of defence response in cAS plants is discussed in this paper. Moreover, in silico analysis revealed that the promoter regions of the genes coding for proteins uniquely accumulating in WT plants shared the CGCG motif, a target of the calcium-calmodulin-binding transcription factor AtSR1 (Arabidopsis thaliana signal responsive1). Therefore, following pathogen perception, the low free cAMP content, altering timing and levels of defence signals, and likely acting in part through the mis-regulation of AtSR1 activity, affected the speed and strength of the immune response.

Background: The olive tree is a typical crop of the Mediterranean basin where it shows a wide diversity, accounting for more than 2,600 cultivars. The ability to discriminate olive cultivars and determine their genetic variability is pivotal for an optimal exploitation of olive genetic resources. Methods: We investigated the genetic diversity within 128 olive accessions belonging to four countries in the Mediterranean Basin (Italy, Algeria, Syria, and Malta), with the purpose of better understanding the origin and spread of the olive genotypes across Mediterranean Basin countries. Eleven highly polymorphic simple sequence repeat (SSR) markers were used and proved to be very informative, producing a total of 179 alleles. Results: Cluster analysis distinguished three main groups according to their geographical origin, with the current sample of Maltese accessions included in the Italian group. Phylogenetic analysis further differentiated Italian and Maltese olive accessions, clarifying the intermediate position of Maltese accessions along the x/y-axes of principal coordinate analysis (PCoA). Model-based and neighbor clustering, PCoA, and migration analysis suggested the existence of two different gene pools (Algerian and Syrian) and that the genetic exchange occurred between the Syrian, Italian and Maltese populations. Discussion: The close relationship between Syrian and Italian and Maltese olives was consistent with the historical domestication and migration of olive tree from the North Levant to eastern Mediterranean basin. This study lays the foundations for a better understanding of olive genetic diversity in the Mediterranean basin and represents a step toward an optimal conservation and exploitation of olive genetic resources., peer-reviewed

The infection of Arabidopsis thaliana plants with avirulent pathogens causes the accumulation of cGMP with a biphasic profile downstream of nitric oxide signalling. However, plant enzymes that modulate cGMP levels have yet to be identified, so we generated transgenic A. thaliana plants expressing the rat soluble guanylate cyclase (GC) to increase genetically the level of cGMP and to study the function of cGMP in plant defence responses. Once confirmed that cGMP levels were higher in the GC transgenic lines than in wild-type controls, the GC transgenic plants were then challenged with bacterial pathogens and their defence responses were characterized. Although local resistance was similar in the GC transgenic and wild-type lines, differences in the redox state suggested potential cross-talk between cGMP and the glutathione redox system. Furthermore, large-scale transcriptomic and proteomic analysis highlighted the significant modulation of both gene expression and protein abundance at the infection site, inhibiting the establishment of systemic acquired resistance. Our data indicate that cGMP plays a key role in local responses controlling the induction of systemic acquired resistance in plants challenged with avirulent pathogens.

Glutathione, a tripeptide constituted by glutamate, cysteine, and glycine, is an abundant metabolite that functions as a master regulator of intracellular redox homeostasis. Under optimal conditions, glutathione is mostly present in the reduced form (GSH), with a free thiol group. The link of two molecules of GSH, via a disulfide bond, leads to the formation of glutathione disulfide (GSSG). GSH can be oxidized, directly or indirectly, by reactive oxygen species, working as a scavenger that prevents excessive oxidation of cellular environment. GSH can also react with different thiols to form mixed disulfides. These reversible redox reactions are responsible for many GSH functions.

In this paper we describe the morphological, histological and chemical characters of 20 native Helichrysum italicum (Roth) G. Don ssp. italicum genotypes collected from different locations in Italy and Corsica (France) and grown under similar edaphic and climatic conditions. An AFLP technique was used to examine the level of genetic variability among the genotypes with the aim to disclose a possible correlation between the genetic and chemical data. The chemical analysis recognizes at least three different chemotypes based on the major constituents in the essential oils. This was confirmed by the AFLP analysis resulting in a dendrogram divided into three main clades. Because of the polymorphic trait of Helichrysum species, an efficient micropropagation protocol was established for the 20 H. italicum (Roth) G. Don ssp. italicum genotypes to guarantee the availability of stable genetic material with defined chemical profiles for industrial applications. The genotypes were found to influence the in vitro performance and the chemical composition of the essential oils, but not the phenotypic traits of the plants.

Olive (Olea europaea L.) is an important fruit species in Italy and Mediterranean basin constituted by a wide germplasm with a large number of cultivars present in all the Mediterranean area. SSRs are becoming the markers of choice for variability studies in olives as they are simple to perform, transferable, hypervariable, highly polymorphic and show a high information content. Olive genetic diversity was studied using 16 SSR markers on 30 cultivars diffused in Southern-Italy. Resolving Power (RP) and Power of Discrimination (PD) were calculated to evaluate the efficiency of the SSR markers investigated in studies of cultivars fingerprinting. Based on their high efficiency, two SSR markers, UDO43 and DCA16 were chosen to set up an identification key to distinguish the entire set of cultivars, confirming the high biodiversity of the Southern-Italian olive germplasm and the suitability of SSR markers in studies of cultivar diagnosis.

The suitability of DNA present in olive oil for PCR analysis has been reported by several authors. However, low concentration, degradation, and the possible presence of additional alleles due to paternal contribution in oils extracted from entire drupes, should be taken into consideration when comparing the amplification profiles of leaves with the corresponding oils for varietal traceability purposes. The aim of this work was to assess, by capillary electrophoresis of microsatellite markers, the phenetic relationships among seven certified Olea europaea L. cultivars, and to verify the genomic equality between leaf DNA and the corresponding monovarietal oil DNA. Moreover, the aim was to establish an identification key to distinguish all the types of oil among them with the minimum number of markers. The results referred to oil DNA, obtained in 70 PCR experiments, confirmed the possibility, in 85.7% of cases, of extracting DNA suitable for the analysis of microsatellites. Ninety percent of the successful amplifications led to identical patterns for leaves and oil DNA. Cima di Melfi shared only 20% of the alleles with the other cultivars. Toscanina and Leccino, showed the highest similarity (about 60%). A single microsatellite was able to distinguish the seven types of oil.

Metschnikowia pulcherrima occurs naturally on fruits, buds and floral parts of apple trees. Some strains are effective as biocontrol agents against postharvest decay of apples and other fruits. The usefulness of the amplified fragment length polymorphism (AFLP) technique was evaluated for the genetic analysis of 26 strains of M. pulcherrima, isolated from different sources in different geographical regions. With six AFLP primer pairs, 729 polymorphic bands were scored. The technique showed a high discriminatory power. Genetic relationships between strains were also estimated using AFLP. All the isolates from the carposphere of apple, previously tested as biocontrol agents, were grouped in a single cluster with a high bootstrap value (97), indicating robustness and reproducibility. AFLP patterns could clearly distinguish the different strains and research is in progress to use some putative specific bands for single tag sequence (STS) conversion to develop isolate-specific markers.

In oil-mills, olive-pomace recentrifugation is a common way to reduce pomace moisture and, at the same time, to recover the oil therein. According to current rules, the obtained oil is defined as “crude olive-pomace oil.” The aim of this work is to verify the effect of recentrifugation on specific chemical and molecular parameters of the crude olive-pomace oil, by comparing it with the corresponding virgin olive oil obtained from the same olive lots. In particular, the following were considered: (i) the polar compounds of the oils that include compounds originated from oxidative and hydrolytic degradation, analyzed by high-performance size exclusion chromatography (HPSEC), and (ii) the profile of DNA microsatellite molecular markers that was analyzed by using the High Resolution Melting (HRM) technique. The obtained results evidenced the significantly higher hydrolytic degradation of crude olive-pomace oil, compared with the corresponding virgin olive oil, but at an extent unlikely able to allow the detection of fraudulent admixtures with virgin olive oils. In addition, the findings demonstrated the feasibility of the application of the HRM analysis of DNA microsatellites to crude olive-pomace oil, able to reveal the alteration of the declared varietal profile of a virgin olive oil sample by simply checking the HRM curve profiles.

Cyclic adenosine 3',5'-monophosphate (cAMP) is a recognized second messenger; however, knowledge of cAMP involvement in plant physiological processes originates primarily from pharmacological studies. To obtain direct evidence for cAMP function in plants, tobacco Bright Yellow-2 (BY-2) cells were transformed with the cAMP sponge, which is a genetically encoded tool that reduces cAMP availability. BY-2 cells expressing the cAMP sponge (cAS cells), showed low levels of free cAMP and exhibited growth inhibition that was not proportional to the cAMP sponge transcript level. Growth inhibition in cAS cells was closely related to the precocious inhibition of mitosis due to a delay in cell cycle progression. The cAMP deficiency also enhanced antioxidant systems. Remarkable changes occurred in the cAS proteomic profile compared with that of wild-type (WT) cells. Proteins involved in translation, cytoskeletal organization, and cell proliferation were down-regulated, whereas stress-related proteins were up-regulated in cAS cells. These results support the hypothesis that BY-2 cells sense cAMP deficiency as a stress condition. Finally, many proteasome subunits were differentially expressed in cAS cells compared with WT cells, indicating that cAMP signaling broadly affects protein degradation via the ubiquitin/proteasome pathway.

Programmed cell death (PCD) is a process of genetically encoded and actively controlled cellular suicide. PCD is associated with different phases of plant life and also represents a defence mechanism that plants activate against different kinds of biotic and abiotic stresses. Reactive oxygen species (ROS) and nitric oxide (NO) have been proposed as key factors in the control of both developmentally and environmentally induced PCD. In this chapter, we give an overview of ROS and NO interplay in the signaling leading to PCD induced in senescence, self-incompatibility, hypersensitive response and under cadmium and heat stress. The data presented indicate that ROS and NO interact through different pathways, depending on the level and the timing of production of these reactive species, the cellular redox state and the plant species. Despite the complicate network of signals, some common points of NO/ROS crosstalk in the different kinds of PCD can be identified. First, during PCD, ROS and NO biosynthesis is mutually regulated with feedback mechanisms. The NO-dependent S-nitrosylation of proteins controlling ROS levels represents a key point of interaction of the two species. Furthermore, NO and ROS interplay could amplify the induction of PCD executors, such as caspase-3-like. Finally, NO and/or ROS might control the ubiquitin–proteasome pathway involved in PCD induction.

Olive oil traceability becomes instrumental to ensure the consumer’s protection, particularly for extra virgin olive oil, the quality of which is highly related to the cultivars employed. The aim of this investigation was to evaluate the possibility of identifying the cultivar used to obtain the derived olive oil by the analysis of amplified fragment length polymorphism (AFLP) markers. To this purpose, ten virgin olive oils were prepared in the laboratory from ten different very common Italian cultivars and were then analysed using six AFLP primer combinations. The technique was optimised for fragmented DNA of oil in order to enhance the intensity of the bands in the AFLP patterns. The obtained results indicated a percentage of polymorphism ranging from 16% for Pst-AGC/Mse-AGT to 43% for Pst-AGC/Mse-ACA. The diversity index was comprised between 90.2% for Pst-AGC/Mse-AGT and 95.2% for Pst-AGG/Mse-AGG. One of the six primer combination analysed, namely Pst-AGG/Mse-AGG, was able to distinguish all the olive oil examined, and a similarity tree of the samples was elaborated.

The aim of the research was to verify the applicability of microsatellite (SSR) markers in High Resolution Melting (HRM) analysis for the identification of the olive cultivars used in the “Terra di Bari” PDO extra virgin olive oil. A panel of nine cultivars, widespread in Apulia region, was tested with seventeen SSR primer pairs and the PCR products were at first analysed with a Genetic Analyzer automatic sequencer. An identification key was obtained for the nine cultivars, which showed an unambiguous discrimination among the varieties constituting the “Terra di Bari” PDO extra virgin olive oil: Cima di Bitonto, Coratina, and Ogliarola. Subsequently, an SSR based method was set up with the DCA18 marker, coupled with HRM analysis for the distinction of the Terra di Bari olive oil from non-Terra di Bari olive oil using different mixtures. Thus, this analysis enabled the distinction and identification of the PDO mixtures. Hence, this assay provided a flexible, cost-effective, and closed-tube microsatellite genotyping method, well suited to varietal identification and authentication analysis in olive oil.

L"Italia possiede il più ampio pool genico di carciofo coltivato, includendo così una ricca biodiversità cinaricola. Le regioni centro-meridionali conservano una moltitudine di tipi varietali ed ecotipi che si differenziano sulla base di tratti morfologici e per composizione chimica, e di conseguenza presentano diverse proprietà nutrizionali e attività farmacologiche. Il carciofo è una rilevante fonte di polifenoli, composti bioattivi dall"elevata capacità antiossidante e dunque molto salutari per l"uomo. Acquisire informazioni sul contenuto e sulla biosintesi di polifenoli in differenti varietà di carciofo, risulta quindi di fondamentale importanza per la promozione e l"utilizzo di risorse genetiche autoctone di questa specie. A tale scopo sono state effettuate dettagliate analisi biochimiche per caratterizzare sei diverse varietà di carciofo (Mola, Tondo di Paestum, Sant"Erasmo, Bianco di Ostuni, Blanca de Tudela e Violetto di Provenza) diffuse non solo in Italia ma anche in altri Paesi del bacino del Mediterraneo. Analisi qualitative e quantitative (HPLC-DAD-ESI-MS/MS) del profilo polifenolico di ciascuna varietà hanno evidenziato che i polifenoli presenti in quantità maggiore sono acido clorogenico, cinarina, luteolin 7-O-rutinoside e luteolin 7-O-glucoside. Inoltre il contenuto varia a seconda del genotipo e del tessuto preso in considerazione (Negro et al. 2012). Uno studio approfondito è stato da tempo avviato su diversi geni coinvolti nella produzione di polifenoli (Sonnante et al. 2008, Sonnante et al. 2010) e, più di recente, sulla regolazione della biosintesi di tali composti nei diversi tessuti di carciofo. Da una libreria di cloni ricombinanti BAC sono stati isolati due geni codificanti per putativi fattori di trascrizione MYB coinvolti nella biosintesi di flavonoli e proantocianidine. Tali geni sono stati caratterizzati strutturalmente. La loro espressione è stata analizzata in diversi tessuti di carciofo tramite PCR quantitativa Real-Time. Le corrispondenti proteine ricombinanti sono state inoltre espresse in batteri, per stabilirne la funzione in vitro tramite lo studio delle interazioni proteine/DNA mediante saggi di mobilità elettroforetica. Bibliografia: Negro et al. (2012) Journal of Food Science 77: 244-252. Sonnante et al. (2008) Physiologia Plantarum 132: 33-43. Sonnante et al. (2010) Plant Physiology 153: 1224-1238.

The ω-3 fatty acid desaturases (FADs) are enzymes responsible for catalyzing the conversion of linoleic acid to α-linolenic acid localized in the plastid or in the endoplasmic reticulum. In this research we report the genotypic and phenotypic variation of Italian Olea europaea L. germoplasm for the fatty acid composition. The phenotypic oil characterization was followed by the molecular analysis of the plastidial-type ω-3 FAD gene ( fad7 ) (EC 1.14.19), whose full-length sequence has been here identified in cultivar Leccino. The gene consisted of 2635 bp with 8 exons and 5′- and 3′-UTRs of 336 and 282 bp respectively, and showed a high level of heterozygousity (1/110 bp). The natural allelic variation was investigated both by a LiCOR EcoTILLING assay and the PCR product direct sequencing. Only three haplotypes were identified among the 96 analysed cultivars, highlighting the strong degree of conservation of this gene.

Southern Italy has geographical, climatic, historical and traditional peculiarities summarized in a landscape to which olive trees are the backdrop. Olive (Olea europaea L. subsp. europaea) is cultivated since the third millennium B.C. in the Eastern region of the Mediterranean sea (Loukas & Krimbas, 1983) and spread later around the basin following land and maritime routes to Italy, Spain, North Africa and France (Angiolillo et al., 1999). The plant longevity, the cross-pollinating nature of the species and its secular history contributed to determine a wide germplasm biodiversity with more than 1200 cultivars, 9.992 thousand harvested hectares and a total production of 18 million tons (Tab.1; FAO 2009) mostly concentrated in the main olive oil producing countries (Alba et al., 2009a). Europe alone produces the 69% of the total worldwide production (Fig. 1). Italy is one of the main producers and has a rich assortment of olive cultivars, with a total number of 395 registered entries in the national list of O. europaea L. Italian cultivars (Official Journal of the Italian Republic, 1994), covering 1,18 million hectares and a total production of 3.6 million tons (ISTAT, 2010). The 80% of the cultivation is mostly concentrated in Southern Italy, where the production touches the 88% of the whole Italian olive production. In particular, Apulia region alone produces 1/3 of the total (about 1,7 milion tons) (Tab. 2). Nowadays olive-growing has a crucial commercial role in many Italian regions with a richness of biodiversity spread from North to South in terms of cultivars grown, agronomic practices, processes of transformation of the raw material. Therefore, the preservation and protection of Italian olive biodiversity depends upon the use of genetic research tools like molecular markers for a correct cultivar identification. In countries that belong to the “Union Internationale pour la Protection des Obtentions Vegetales” (UPOV) the registration, multiplication, certification, and commercialization of olive varieties rely on the evaluation of morpho-agronomic descriptors. Recently, morphological, biochemical and agronomic traits have been complemented with the large array of DNA molecular marker types

Background Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource. Results To balance the maximum mutation density with an acceptable plant survival rate, a 'kill curve' analysis was first conducted with different ethylmethanesulfonate (EMS) dosages and different exposure times. According to the germination rate, a treatment with 0.7% EMS for 6 h was chosen. An M2 progeny of 3,651 fertile plants was obtained. Totally, 4.79% of the whole population showed clear aberrant phenotypes. A microsatellite analysis on a representative sample of the original seed stock and mutant lines confirmed the uniformity of the genetic background of plant material. The TILLING procedure was successfully applied to sunflower genome, initially by a CelI-nuclease mismatch cleavage assay coupled with a DNA-pooling level test. To investigate the efficiency of the mutagenic treatment, a pilot screening was carried out on 1,152 M2 lines focusing on four genes, three involved in the fatty acid biosynthetic pathway and one for downy mildew resistance. A total of 9 mutant lines were identified and confirmed by sequencing; thereby, the estimated overall mutation frequency for the pilot assay resulted to be 1/475 kb. Conclusion A first TILLING population for a high throughput identification of EMS-induced point mutations in sunflower genome has been successfully obtained. This represents a powerful tool to a better understanding of gene function in sunflower.

The rationale for the following research was to analyse the response of sunflower hybrids to different sowing dates and to evaluate hybrid response to critical environmental conditions. The data used are from an experiment conducted in a location-year combination over a period of two years (2007–09) in southern Italy. Eleven hybrids were tested following a randomized complete block design with three replications at each location-year combination. Eight agronomic characters including seed oil content were recorded. Classification and ordination procedures were used to investigate hybrid performance in relation to three different sowing dates. Combined analysis of variance showed that hybrids, location-year combination, sowing date and their interactions were highly significant for all characters. Hybrid performances were classified by cluster analysis into groups that were differentiable in terms of means and stability. The first three components accounted for 74%, 82%, and 87% of the total variation for the first, second and third sowing date respectively. Plotting component one against component two along Euclidean axes should therefore provide a reasonable representation of the spatial arrangements of hybrid performances in the original multi-dimensional space. The applied statistical method gives full information on hybrid performances similarity.

Italian industrial pasta and durum wheat typical breads must be prepared using exclusively durum wheat semolina. Previously, a microsatellite sequence specific of the wheat D-genome had been chosen for traceability of soft wheat in semolina and bread samples, using qualitative and quantitative Sybr green-based real-time experiments. In this work, we describe an improved method based on the same soft wheat genomic region by means of a quantitative real-time PCR using a dual-labeled probe. Standard curves based on dilutions of 100% soft wheat flour, pasta, or bread were constructed. Durum wheat semolina, pasta, and bread samples were prepared with increasing amounts of soft wheat to verify the accuracy of the method. Results show that reliable quantifications were obtained especially for the samples containing a lower amount of soft wheat DNA, fulfilling the need to verify labeling of pasta and typical durum wheat breads.

Collina di Brindisi is an Italian extra virgin olive oil that obtained the mark of protected designation of origin (PDO) according to EC Regulation 2081/92. The varietal requirements of the official production protocol of this oil foresee that this oil is prepared from cultivar Ogliarola (minimum 70%) and other Olea europaea L. cultivars that are diffused in the production area, accounting for a maximum of 30%. The aim of this work was to verify the effectiveness of microsatellite analysis in verifying the identity of Collina di Brindisi PDO olive oil. A preliminary assessment of product's quality by means of chemical analyses was also carried out. Microsatellite analysis clarified that the generic name Ogliarola, indicated in the technical sheet of this PDO oil, actually corresponded to the Ogliarola salentina cultivar. Furthermore, the obtained results showed that the examination of a limited number of DNA microsatellites enables the identification of the Ogliarola salentina cultivar in this PDO oil.