The Development Of EST-SSR Marker And Genetic Relationship In Vitis Adenoclada Hand. -Mazz

Vitis adenoclada Hand.-Mazz is the China's unique kinds of wild grapes,which widely distribute in the areas out of the south of Changjiang River,such as Hunan, and so on. In the study, taking these grapes growing in the teach base of HunanAgriculture University and Zhengzhou Fruit Research Institute for materials, Preliminary observations and record characteristics of its morphological characters.Also,we study the phylogenetic relationship among v.adenoclada Hand.-Mazz and several other wild grae species and cultivars by using EST-SSR marker and clustering analysis. The main results are as below:(1)Adopted materials by 19 grape leaves according to 8 traits were clustered, there were significant differences in most morphological traits among populations. Fome 0.0705 to 2.3762.(2)Established a effective and reproductive PCR reaction system for EST-SSR. Each 25μl PCR:30ng of DNA,2.5mMof Mgcl20.2μMof each primer,10mM of Tris-Hcl,50 mMof KCl,lunit of Amplitaq Gold and 0.2mMof dNTPs,and they were cycled at 94℃for 1 min,50℃for 1 min,and 73℃for 1 min repeated for 45 cycles.(3)Adopted large morphological differences grapes,8 pairs of primers were selected from 16 pairs of EST-SSR primers which produced the whole of 49 bands,of which 37 were polymophic with a polymophicsm of 75.5%.(4)Study of 23 grape and EST-SSR related species for analysis,clustering resultsshow that:similar to coefficient of 0.8 for the critical value, can be divided into 8 groups.(5)The test on the gland at the molecular level of the relationship between the classification of Vitis adenoclada Hand.-Mazz.The results showed that the genetic coefficient (0.9556)bewteen Vitis quinquangularis Rehd.It Shows more recent genetic relationship,So it verified the Taxonomic result made by Niu Lixin.