Studies On Phylogenetic Relationship And Analysis Of Grape In Germplasm Resource By RAPD

Owing of the variation within the species,easily nature cross between species and wide generation,different planting environment in the world,many species populations and variety groups of Grape(vitis) have been forming in the long history of grape evolution and culturing,which make taxonomy and identification of grapes difficult and naming confusing.The study was undertaken to identify grape(vitis) material of several grapes of V.vinifera L. and V.labrusca L. and rootstock by RAPD. This paper mainly studies the genetic diversity of 53 series grapes on RAPD. The aim of the studies on the amplification patterns of RAPD got by NTSYS pc2.10e is to identificate and taxonomy grape,study linkage distance.The main results as below:1 .The methods-CTAB method,SDS method,and high salinity and low PH method were used to extract genomic DNA from leaves of grape.The DNA samples of the three methods were tested by spectrophotometer,agarose gelelectrophoresis and RAPD.The result was that the DNA integrality extract with the three methods was good,the DNA trap was distinct,the number of A260/A280 lied between 1.7 and 2.0,the DNA could conform to the request,which could be used for extracting genomic DNAfrom leaves of grape,and that the CTAB method was the best ones through which the production had the highest purity.So,we chose CTABmethod to conduct following experiments.2.The different PCR program and the ingredients in RAPD reation system such as DNA and primer ,Taq DNA polymerase, dNTP,Mg2+, were sceened by different concentrations. Suitable reaction system and program of the RAPDfor grape genomic DNAwas settled.The volumn of amplified reation was 25ul, containing10× buffer (PH=8. 8) 2.5ul, genomic DNA 50ng, 0.4umol/L primer, 200umol/L dNTP, 1U Taq DNA polyrase 2. 0mM Mg2+,and pure water 15.5ul.The reaction program was devised for 39 cycles,each with 94℃ denaturation for 1 min,36℃ annealing for 1 min ,72℃ extension for 2 min,and 94℃ predenature for 2 min in the former denature and 72℃ postextension for 5 min in the final extension.3. 33 primers which are the better of 280 primers in the experiment are used to analyze the genetic diversity of 53 series grapes using RAPD markers. A total of 280...