Establishment Of SRAP Reaction System And Analysis Of Genetic Diversity Of Grape Cultivars

Grape is one of the world’s most widely cultivated fruit trees of one.During long history of evolutionand grapevine growth,many specie groups and variety groups has formed.Because aberrance in specieoccurres all along and grapevines are propagated by cutting,so these often make grape varietiesidentification difficult.In this paper in the establishment and optimization of grapes of a molecular markerreaction system, based on the application of SRAP molecular markers for156grape germplasm wereanalyzed, and the evaluation of selected grapes the genetic relationship and genetic diversity of germplasmresources.The main results from this study are as follows:1. Compared three methods of DNA extraction,DNA sample extracted by these three methods were allof good quality for SRAP. Comparatively,method of the modified CTAB was simpler,more efficient andDNA sample obtained has higher purity.The result showed that the modified CTAB method found in thispaper could cffectively eliminate the affections of the secondary materials to extracted genome DNA fromgrape leaves,and which be suitable for SRAP analysis．2. Constructed the SRAP reaction system,respectively.The best SRAP reaction mixtures had a totalvolume of25ul, which containing1μL templateDNA(50ng)1μL, primer （10μmol·L-1）,0.2μL TaqDNAPolymerase，（2.5U·μL-1），0.5μL dNTPs（10mmol·L-1）。3. From100pairs of SRAP can extract the highly polymorphic primers, stable and clear amplificationof10primers, the156grape germplasm genetic diversity analysis, the results show that by10primeramplification out174bands, including polymorphism with173, the polymorphism proportion is99.43%.Using POPGEN version1.32software to calculate the selected grapes of Shannon index was0.4434~0.5859, the range of Nei’s genetic diversity for the range of0.2863~0.4062.4. Use NTSYSpc2.10t software on156samples of grapes of a tag amplification results werestatistically analyzed, grape samples between the genetic similarity coefficient variation between0.259~1.000, by the weighted average (UPGMA) set the rule of law as a genetic relationship between tree, SRAPmarkers in the similarity coefficient of0.522will all the materials into three categories, respectively.