A study of genetic profiles of grape plants preserved under the name of Dagestan variety ‘Khatmi’

E. T.  Ilnitskaya, M. V. Makarkina, R. E. Kazahmedov, E. A. Kozhevnikov, T. D. Kozina
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Abstract

Background . Traditionally, the description of grape varieties is a task of ampelographic studies. However, several different grape cultivars have similar phenotypic traits. Molecular genetic characterization is the most accurate tool for cultivar identification. The development of DNA fingerprinting of varieties is the first step in this direction. An extensive database of DNA profiles of grape genotypes makes it possible to determine the varietal affiliation of unknown forms, confirm or refute the varietal correspondence of planting material. ‘Khatmi’ is an autochthonous grape variety from Dagestan. The profile of ‘Khatmi’ is presented in the V IVC international database of DNA fingerprints for grape varieties. However, an application of DNA analysis methods in grape variety studies has determined that several ancient varieties were cultivated under one name, while for others a certain variability of genotypes was observed. The objectives of the work were to study samples of ‘Khatmi’ plants from different places of growth in Dagestan by standard microsatellite loci used for grape genotyping, to assess the level of genetic similarity of the samples, and to refine the DNA profile of ‘Khatmi’. Materials and methods. Molecular genetic study was carried out on 10 samples from different ‘Khatmi’ populations. The material was picked from the collection sites of Dagestan breeding experimental station of viticulture and vegetable growing and the Dagestan Experiment Station of VIR, as well as from production plantations. DNA was extracted from herbarium specimens of young grape shoot tips by the CTAB method. The samples were genotyped by polymerase chain reaction using a standard set of primers for 9 microsatellite markers VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62 and VrZAG79 recommended by the International Organization of Grapes and Wine (OIV) for grapevine DNA fingerprinting. Amplification products were separated, and their sizes were assessed using capillary electrophoresis on an ABI Prism 3130 genetic analyzer. Results. Genotyping was done for 10 samples of grapes growing in Dagestan under the name ‘Khatmi’, including samples from different collections and places of industrial cultivation, as well as clonal variations of this variety and putative clonal variations. The two base pair differences within one of the loci distinguished the DNA profiles of the analyzed samples from that of ‘Khatmi’ presented in the international grape varieties database V IVC. It was determined that the sample under the name ‘Khatmi krupnoyagodnyi’ is closely related to ‘Khatmi’ variety by its genotype, but probably represents a clonal variation of ‘Koz uzyum’, another local variety of Dagestan. Conclusion. The DNA profile of the local Dagestan grape variety ‘Khatmi’ has been refined.
达吉斯坦葡萄品种Khatmi的遗传图谱研究
背景。传统上,葡萄品种的描述是地形学研究的一项任务。然而,几种不同的葡萄品种具有相似的表型性状。分子遗传鉴定是品种鉴定最准确的工具。品种DNA指纹图谱的开发是这一方向的第一步。一个广泛的葡萄基因型DNA图谱数据库,可以确定未知形式的品种归属,证实或反驳种植材料的品种对应。Khatmi是达吉斯坦的一种本地葡萄品种。“Khatmi”的基因图谱已在vivc国际葡萄品种DNA指纹数据库中公布。然而,DNA分析方法在葡萄品种研究中的应用表明,几个古老的品种是在一个名称下栽培的,而对于其他品种,则观察到一定的基因型变异性。这项工作的目标是通过用于葡萄基因分型的标准微卫星位点研究来自达吉斯坦不同生长地点的“Khatmi”植物样本,评估这些样本的遗传相似性水平,并完善“Khatmi”的DNA图谱。材料和方法。对来自不同“哈特米”种群的10个样本进行了分子遗传学研究。材料采自达吉斯坦葡萄栽培和蔬菜种植育种实验站和VIR达吉斯坦实验站的采集点,以及生产种植园。采用CTAB法从植物标本室葡萄幼芽尖标本中提取DNA。采用国际葡萄与葡萄酒组织(OIV)推荐的9种葡萄DNA指纹图谱微卫星标记(VVS2、VVMD5、VVMD7、VVMD25、VVMD27、VVMD28、VVMD32、VrZAG62和VrZAG79)的标准引物,采用聚合酶链反应对样品进行基因分型。扩增产物分离,在ABI Prism 3130基因分析仪上进行毛细管电泳测定。结果。对在达吉斯坦种植的10个以“Khatmi”命名的葡萄样本进行了基因分型,包括来自不同采集地和工业栽培地的样本,以及该品种的克隆变异和假定的克隆变异。其中一个位点内的两个碱基对差异将分析样品的DNA图谱与国际葡萄品种数据库V IVC中的“Khatmi”区分开来。经确定,以“Khatmi krupnoyagodnyi”命名的样品与“Khatmi”品种的基因型密切相关,但可能代表达吉斯坦另一个当地品种“Koz uzyum”的克隆变异。结论。达吉斯坦当地葡萄品种“Khatmi”的DNA图谱已经得到了改进。
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