Milovanov Alexander, Zvyagin Andrey, Daniyarov Asset, Kalendar Ruslan, Troshin Leonid
Department of Viticulture, Kuban State Agrarian University, Krasnodar, 350044, Russia.
Laboratory of Plants Genomics and Bioinformatics, RSE "National Center for Biotechnology", Astana, 010000, Kazakhstan.
Genetica. 2019 Feb;147(1):91-101. doi: 10.1007/s10709-019-00055-5. Epub 2019 Feb 19.
Cultivated grapevine (Vitis vinifera L. ssp. sativa D.C.) is one of the oldest agricultural crops, each variety comprising an array of clones obtained by vegetative propagation from a selected vine grown from a single seedling. Most clones within a variety are identical, but some show a different form of accession, giving rise to new divergent phenotypes. Understanding the associations among the genotypes within a variety is crucial to efficient management and effective grapevine improvement. Inter-primer binding-site (iPBS) markers may aid in determining the new clones inside closely related genotypes. Following this idea, iPBS markers were used to assess the genetic variation of 33 grapevine genotypes collected from Russia. We used molecular markers to identify the differences among and within five grapevine clonal populations and analysed the variation, using clustering and statistical approaches. Four of a total of 30 PBS primers were selected, based on amplification efficiency. Polymerase chain reaction (PCR) with PBS primers resulted in a total of 1412 bands ranging from 300 to 6000 bp, with a polymorphism ratio of 44%, ranging from 58 to 75 bands per group. In total, were identified seven private bands in 33 genotypes. Results of molecular variance analysis showed that 40% of the total variation was observed within groups and only 60% between groups. Cluster analysis clearly showed that grapevine genotypes are highly divergent and possess abundant genetic diversities. The iPBS PCR-based genome fingerprinting technology used in this study effectively differentiated genotypes into five grapevine groups and indicated that iPBS markers are useful tools for clonal selection. The number of differences between clones was sufficient to identify them as separate clones of studied varieties containing unique mutations. Our previous phenotypic and phenological studies have confirmed that these genotypes differ from those of maternal plants. This work emphasized the need for a better understanding of the genotypic differences among closely related varieties of grapevine and has implications for the management of its selection processes.
栽培葡萄(Vitis vinifera L. ssp. sativa D.C.)是最古老的农作物之一,每个品种都包含一系列通过营养繁殖从单株实生苗培育出的选定葡萄藤获得的克隆。一个品种内的大多数克隆是相同的,但有些表现出不同的类型,产生新的不同表型。了解一个品种内基因型之间的关联对于高效管理和有效改良葡萄至关重要。引物结合位点间(iPBS)标记可能有助于确定亲缘关系密切的基因型内的新克隆。基于这一想法,iPBS标记被用于评估从俄罗斯收集的33个葡萄基因型的遗传变异。我们使用分子标记来识别五个葡萄克隆群体之间和内部的差异,并使用聚类和统计方法分析变异。根据扩增效率,从总共30个PBS引物中选择了4个。用PBS引物进行聚合酶链反应(PCR)共产生1412条带,大小在300至6000 bp之间,多态性比率为44%,每组有58至75条带。在33个基因型中总共鉴定出7条私有带。分子方差分析结果表明,总变异的40%出现在群体内,仅60%出现在群体间。聚类分析清楚地表明,葡萄基因型高度分化且具有丰富的遗传多样性。本研究中使用的基于iPBS PCR的基因组指纹技术有效地将基因型分为五个葡萄组,并表明iPBS标记是克隆选择的有用工具。克隆之间的差异数量足以将它们识别为含有独特突变的研究品种的不同克隆。我们之前的表型和物候学研究证实,这些基因型与母本植物不同。这项工作强调了更好地理解葡萄亲缘关系密切的品种间基因型差异的必要性,并对其选择过程的管理具有启示意义。