葡萄砧木品种的SSR分析  

Analysis of grape rootstocks by SSR markers

作　　者：王雯染 杨哲[1] 杨航宇 王军[1,2] 

机构地区：[1]中国农业大学食品科学与营养工程学院葡萄与葡萄酒研究中心,北京100083 [2]农业部葡萄酒加工重点实验室,北京100083

出　　处：《果树学报》2018年第1期11-19,共9页Journal of Fruit Science

基　　金：现代农业产业技术体系建设专项基金(CARS-29)

摘　　要：【目的】分析葡萄砧木品种遗传多样性。【方法】利用17对SSR引物对21个葡萄砧木品种、3个欧亚种酿酒葡萄品种进行扩增。【结果】共获得195个等位基因,片段大小为87 bp(VVIV52)~265 bp(Vrzag79),观测杂合度(Ho)为0.46~0.96,期望杂合度(He)为0.57~0.94,Shannon多样性指数(I)为1.10~2.62,多态性百分率为100%,所选引物的遗传多态性高,适宜用作葡萄砧木品种的遗传多样性分析。在单个引物鉴定品种方面,VVMD5效率最高,可鉴定19个品种;VVS3和UDV-058效率最低,仅能鉴别2个品种。选择2对引物组合可以大大提高鉴定效率,选择VVMD5&VVS4等组合可将全部供试种质资源鉴定出来,鉴定效率达100%,体现了SSR标记在品种鉴定上的高效性。对扩增结果进行聚类分析,在遗传相似系数0.74处,欧亚种酿酒葡萄与砧木品种各自聚为一类,且亲缘关系较近的砧木品种聚在一起,聚类结果符合砧木品种选育时的亲缘关系。【结论】利用SSR分子标记能有效地鉴定葡萄砧木品种和进行遗传多样性分析。[Objective] The rootstock varieties of grape commonly used in China are mostly selected from Europe and America, some rootstock varieties have been confused during the course of introduction. Moreover, many grape rootstock varieties have similar genetic background and similar morphology, which are difficult to identify by traditional morphological characters. Therefore, the correct identification of grape rootstock varieties is of great significance for efficient utilization and protection of the rootstock resources. Simple sequence repeat （SSR） is a molecular genetic marker based on PCR technology. SSR marker has been widely used in genetic identification, relationship analysis, genetic map construction, and pedigree reconstruction of grape germplasm resources for its advantages of high polymorphism, good repeatability, codominant inheritance and strong specificity. This study was carried ou to analyze the genetic relation- ship and genetic diversity of grape rootstocks. [ Methods ] 21 grape rootstocks and 3 varieties of Vitis vinifera varieties ‘Cabernet Sauvignon＇ ‘Chardonnay＇ ‘Syrah＇ were amplified by 17 pairs of SSR primers, such as VVS2, VVMD5, VVMD7, VVMD27, Vrzag62, Vrzag79 and so on. Genomic DNA of grape leaf samples was extracted by modified CTAB method. Genomic SSR-PCR was amplified using the following amplification systems and procedures： The system included 2 μL, 10×PCR buffer, 1 U Taq enzyme, 0.2 mmol·L^-1 dNTP, 1.5 mmol·L^-1 Mg^2＋, 0.4 μmol·L^-1 primer （each）, 50 ng DNA template, and ddH20 supple- mentation to 20 μL. The PCR reaction was carried out on the Prime G02 PCR instrument produced by Bibby Scientific. The running amplification reaction program was as follows： an initial denaturation 94℃ for 5 min, denaturation at 94 ℃ for 30 s, annealing at 50-60 ℃ for 30 s, extension at 72℃ for 40 s, reaction of 35 cycles, last extension at 72 ℃ for 10 min. The same template was repeated twice. SSR-PCR am- plification products were analyzed by 8% denatured polyac

关 键 词：葡萄砧木 SSR分子标记 品种鉴定 遗传多样性 

分 类 号：S663.1[农业科学—果树学]