Nicolè Silvia, Barcaccia Gianni, Erickson David L, Kress John W, Lucchin Margherita
Laboratory of Plant Genetics and Genomics, DAFNAE, University of Padova, Campus of Agripolis - Viale Università 16, 35020 Padova, Legnaro, Italy.
BMC Res Notes. 2013 Dec 3;6:502. doi: 10.1186/1756-0500-6-502.
Vitis vinifera L. is one of society's most important agricultural crops with a broad genetic variability. The difficulty in recognizing grapevine genotypes based on ampelographic traits and secondary metabolites prompted the development of molecular markers suitable for achieving variety genetic identification.
Here, we propose a comparison between a multi-locus barcoding approach based on six chloroplast markers and a single-copy nuclear gene sequencing method using five coding regions combined with a character-based system with the aim of reconstructing cultivar-specific haplotypes and genotypes to be exploited for the molecular characterization of 157 V. vinifera accessions. The analysis of the chloroplast target regions proved the inadequacy of the DNA barcoding approach at the subspecies level, and hence further DNA genotyping analyses were targeted on the sequences of five nuclear single-copy genes amplified across all of the accessions. The sequencing of the coding region of the UFGT nuclear gene (UDP-glucose: flavonoid 3-0-glucosyltransferase, the key enzyme for the accumulation of anthocyanins in berry skins) enabled the discovery of discriminant SNPs (1/34 bp) and the reconstruction of 130 V. vinifera distinct genotypes. Most of the genotypes proved to be cultivar-specific, and only few genotypes were shared by more, although strictly related, cultivars.
On the whole, this technique was successful for inferring SNP-based genotypes of grapevine accessions suitable for assessing the genetic identity and ancestry of international cultivars and also useful for corroborating some hypotheses regarding the origin of local varieties, suggesting several issues of misidentification (synonymy/homonymy).
葡萄(Vitis vinifera L.)是社会上最重要的农作物之一,具有广泛的遗传变异性。基于葡萄形态学特征和次生代谢产物识别葡萄基因型存在困难,这促使人们开发适用于品种遗传鉴定的分子标记。
在此,我们对基于六个叶绿体标记的多位点条形码方法与使用五个编码区域结合基于特征系统的单拷贝核基因测序方法进行了比较,目的是重建品种特异性单倍型和基因型,以用于157份葡萄品种的分子特征分析。叶绿体目标区域的分析证明了DNA条形码方法在亚种水平上的不足,因此进一步的DNA基因分型分析针对所有品种扩增的五个核单拷贝基因的序列。UFGT核基因(UDP-葡萄糖:类黄酮3-O-葡萄糖基转移酶,浆果表皮中花青素积累的关键酶)编码区域的测序发现了判别性单核苷酸多态性(1/34 bp),并重建了130个葡萄不同的基因型。大多数基因型被证明是品种特异性的,只有少数基因型被更多(尽管密切相关)的品种共享。
总体而言,这项技术成功地推断出了基于单核苷酸多态性的葡萄品种基因型,适用于评估国际品种的遗传身份和谱系,也有助于证实一些关于当地品种起源的假设,提示了一些误认(同义/同名)问题。