Chloroplast SSR markers to assess DNA diversity in wild and cultivated grapevines

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Genetic relationships among grapevine cultivars grown in Oltrepò Pavese (Italy)

In order to evaluate the genetic distance among 47 grapevine accessions, including major and minor local cultivars grown in Oltrepò pavese (Pavia, Italy), DNA has been analysed with the AFLP approach (VOS et al. 1995). The electrophoretic analysis of the products amplified by 3 primer combinations showed high polymorphism.Furthermore, considering that DNA markers for haploid, uniparentally inherited genomes, such as the chloroplast DNA, are important indicators of pedigree, CpSSR analysis was also performed and this identified cases of maternal common origin among the analysed cultivars. The results show that these molecular tools allow univocal genotype identification and that the analysed germplasm has a wide genetic dissimilarity. Results are coherent with the postulation of a polycentric origin for the Oltrepò pavese cultivars and of a multiple varietal flow from different viticultural regions. The molecular information gathered in this research is essential for the establishment of an appropriate presentation programme of autochthonous varieties

Uptake and effects of a mixture of widely used therapeutic drugs in Eruca sativa L. and Zea mays L. plants

Pharmaceutically active compounds (PACs) are continuously dispersed into the environment due to human and veterinary use, giving rise to their potential accumulation in edible plants. In this study, Eruca sativa L. and Zea mays L. were selected to determine the potential uptake and accumulation of eight different PACs (Salbutamol, Atenolol, Lincomycin, Cyclophosphamide, Carbamazepine, Bezafibrate, Ofloxacin and Ranitidine) designed for human use. To mimic environmental conditions, the plants were grown in pots and irrigated with water spiked with a mixture of PACs at concentrations found in Italian wastewaters and rivers. Moreover, 10 7 and 100 7 concentrations of these pharmaceuticals were also tested. The presence of the pharmaceuticals was tested in the edible parts of the plants, namely leaves for E. sativa and grains for Z. mays. Quantification was performed by liquid chromatography mass spectroscopy (LC/MS/MS). In the grains of 100 7 treated Z. mays, only atenolol, lincomycin and carbamazepine were above the limit of detection (LOD). At the same concentration in E. sativa plants the uptake of all PACs was >LOD. Lincomycin and oflaxacin were above the limit of quantitation in all conditions tested in E. sativa. The results suggest that uptake of some pharmaceuticals from the soil may indeed be a potential transport route to plants and that these environmental pollutants can reach different edible parts of the selected crops. Measurements of the concentrations of these pharmaceuticals in plant materials were used to model potential adult human exposure to these compounds. The results indicate that under the current experimental conditions, crops exposed to the selected pharmaceutical mixture would not have any negative effects on human health. Moreover, no significant differences in the growth of E. sativa or Z. mays plants irrigated with PAC-spiked vs. non-spiked water were observed

Genomic stability in Arabidopsis thaliana transgenic plants obtained by floral dip

. The occurrence of DNA modification is an undesired phenomenon accompanying plant cell transformation. The event has been correlated with the stress imposed by the presently utilised transformation procedures, all depending on plant differentiation from in vitro cell culture, but other causes have not been excluded. In this work, transgenic Arabidopsis thaliana plants have been produced by an approach that does not require cell dedifferentiation, being based on in planta Agrobacterium-mediated gene transfer by flower infiltration, which is followed by recovery and selection of transgenic progeny. Genomic DNA changes in transgenic and control plants have been investigated by AFLP and RAMP analysis. Results show no statistically relevant genomic modifications in transgenic plants, as compared with control untreated plants. Variations were observed in callus-derived A. thaliana plants, thus supporting the conclusion that somaclonal variation is essentially correlated with the stress imposed by the in vitro cell culture, rather than with the integration of a foreign gene

Phylogenetic analysis of grapevine cv. Ansonica growing on the island of Giglio, Italy, by AFLP and SSR markers

The geographic origin of the grapevine cultivar Ansonica (Vitis vinifera L.) grown on the island of Giglio, Tuscany, Italy, was investigated with molecular tools, i.e. AFLP and SSR (microsatellite) analysis. The scored polymorphic DNA bands were statistically analysed and the results were expressed in dendrograms showing the degree of genomic similarity among the tested cultivars: three Ansonica vines sampled in some of the oldest vineyards of the island, one Ansonica specimen grown in Tuscany, one Inzolia specimen grown in Sicily, 23 cultivars grown in different Mediterranean regions and 40 Greek cultivars. The results obtained from AFLP or SSR approaches led to equivalent conclusions: the three grapevines sampled in Giglio were indistinguishable and showed genetic similarity with cv. Ansonica grown in the Tuscan mainland and with cv. Inzolia from Sicily, Airen from Spain, Clairette from France and Roditis from Greece. The SSR analysis showed that Sideritis and Roditis have the highest genomic similarity with Ansonica among the 40 tested Greek cultivars. The molecular analysis gives conclusive evidence for the Greek origin of Ansonica grown on the island, as previously proposed on the basis of morphological and historical studies

Genetic studies on Trebbiano and morphologically related varieties by SSR and AFLP markers

Several grapevine varieties are named Trebbiano and have similar phenotypical characteristics (e.g. whitish berries, late ripening, vigorous growth). Other vines share many of the morphological characteristics of Trebbiano but have different names. In order to clarify the relationships between some of these varieties, a study was carried out with 7 Trebbiano varieties and 17 cultivars that are morphologically similar to Trebbiano. AFLP and SSR analyses were performed to define genetic similarity among the Trebbiano cultivars as well as between the Trebbiano cultivars and related varieties. Results show a large degree of variability between most of the cultivars and suggest that they do not share a common ancestor. Genetic analysis revealed that cv. Verdicchio is identical with Trebbiano di Soave which is very similar to Trebbiano di Lugana.

Phylogenetic analysis of grapevine cv. Ansonica growing on the island of Giglio, Italy, by AFLP and SSR markers

The geographic origin of the grapevine cultivar Ansonica (Vitis vinifera L.) grown on the island of Giglio, Tuscany, Italy, was investigated with molecular tools, i.e. AFLP and SSR (microsatellite) analysis. The scored polymorphic DNA bands were statistically analysed and the results were expressed in dendrograms showing the degree of genomic similarity among the tested cultivars: three Ansonica vines sampled in some of the oldest vineyards of the island one Ansonica specimen grown in Tuscany, one Inzolia specimen grown in Sicily, 23 cultivars grown in different Mediterranean regions and 40 Greek cultivars. The results obtained from AFLP or SSR approaches led to equivalent conclusions: the three grapevines sampled in Giglio were indistinguishable and showed genetic similarity with cv. Ansonica grown in the Tuscan mainland and with cv. Inzolia from Sicily, Airen from Spain, Clairette from France and Roditis from Greece. The SSR analysis showed that Sideritis and Roditis have the highest genomic similarity with Ansonica among the 40 tested Greek cultivars. The molecular analysis gives conclusive evidence for the Greek origin of Ansonica grown on the island, as previously proposed on the basis of morphological and historical studies

Biodiversity of grapevines (Vitis vinifera L.) grown in the Aosta Valley

The PCR-based DNA microsatellite analysis has been applied to define genetic relationships among 19 most representative grapevine cultivars in the Aosta valley. Eighteen varieties grown nearby, in the Savoy (France), Valais (Switzerland) and Piedmont (Italy) regions, were also analysed, to verify the correlations with the Aosta cultivars inferred by the analysis of synonyms and/or morphology. Results were obtained by analysing 12 DNA microsatellite loci. High biodiversity has been recorded amongst the analysed grapevines. In some cases cultivars known with different names in the Valley and in the nearby regions displayed the same SSR allele size, proving the occurrence of synonyms. In other case, identical denominations did not correspond with the genomic identity.