Genetic Diversity In Grape Varieties Based On Randomly Amplified Polymorphic DNA (RAPD) Analysis

Vitis vinifera L. is the predominant commercial specie in all of Vitaceae Lindl plants, including many high-grade varieties (over 5000). Most of high quality table grapes,wine grapes and other processing varieties are belong to this specie or its hybrids. During long history of evolution and grapevine growth, many specie groups and variety groups has formed. Because aberrance in specie occurres all along and grapevines are propagated by cutting, so these often make grape varieties identification difficult. Genetic diversity of 68 grape varieties and 21grape clones has been researched and the results are as flows: I. Compared two methods of DNA extraction:SDS and CTAB. DNA sample extracted by these two methods were all of good quality for RAPD. Comparatively, the method of CTAB was simpler,more efficient and DNA sample obtained has higher purity. II. Optimized RAPD reaction system and suitable conditions was obtained: 1×buffer, 0.2mM dNTP, 2.0mM Mg²⁺, 1.5U Taq DNA polymerase, 0.2μM primer, 20～40ng template DNA;reaction condition:94 ℃7min; 94℃1min,36℃1min,72 ℃2min,45 cycles; 72℃,10min。III. 56 primers were used for amplification, among which 18 primers were selected for RAPD analysis. These primers could amplify stable and polymorphic RAPD bands. IV. Tree diagram was constructed based on Euclidean distance matrix using UPGMA (unweighted pair group method using arithmetic average) cluster analysis. It was showed on the dendrogram that 68 varieties belong to 9 groups: V. davidii Foex is the first group; V. amurensis Rupr. is the second group; V. quinquangularis Rehb is the third group; Zana, Jingxiu, Muscat Hamburg, Zao jinxiang, Zao Meigui, Queen of Vineyard, Shandong Zhaohong and so on (8 varieties) formed fourth group;Sauvignon Blanc, Gewurztraminer, Carignan, Riesling, Chenin Blanc, Ugni Blanc and so on (18 varieties) formed fifth group, these blong to Proles occidentalis Negr; Zexiang, Baijixin, Kuibao, Feihong, Rizamat, Manaizi, Blush Seedless and so on (16 varieties) formed sixth group, these blong to Proles orientalis Negr; Red Gamay, Muscat Rose, Camay Blanc, Muscat Blanc (4 varieties) formed seventh group, these blong to Proles pontica Negr ; Tensyu, Blank Olympia, Beniizu, Shina bailey, Kuroshio, Zi Zhenxiang, Campbell Early, Maciji, Honey Red, Ikawa 1014, Ikawa 1025, Benizuiho, Jingya and so on (13 varieties) formed eighth group; Kyoho, Sato, Jingchao, White fuji (4 varieties) formed ninth group. V. Using morphological method and RAPD method respectively to clustering analysis 21 grape clones, the results showed that only using morphological method can't distinguish these clones, but using RAPD method can distinguish them. clustering analysis indicated that Cabernet Franc, Cabernet Sauvignon and Molet have close relationship.