Studies On The Application Of RAPD Molecular Markers To The Relationship And Classification Of Watermelon Materials

In this study, the genetic diversity of 42 watermelon materials from Sichuan Agricultural University were studied, Pass the covariance analysis of the biology form that the in the field, indoor inquisition carries on the watermelon; Adoption the improvement CTAB method to extract watermelon genomic DNA; Established the optimal RAPD-PCR system in watermelon; The sieving is applicable to experiment originally of primer; Compute the genetic distance between materials and combines to carry on gathering an analysis to the material. The results as following:1,Carry on the observation covariance through in the field and indoor 42 biologies of watermelons materials of rightness forms. The results showed that these materials at the period of pour rattan , bloom the period and sit the fruit stanza, the result number, plant diseases and insect pests, fruit type, fruit heavy, the fruit peel degree of hardness, fruit peel color, flesh of fruits color, fruit peel is thick, flesh boundary, the seed size how much color, TSS etc. all exist the certain difference2,The improved CTAB method developed in this study was the best one to extract watermelon genomic DNA. DNA quality was high, and OD260/OD280 of DNA was 1.8-1.9.And DNA without getting rid of RNA could be used for RAPD amplification.3,The optimal RAPD-PCR system in watermelon was established with orthogonal design.In a total volume of 25μL RAPD-PCR system ,it contains 2.5μL 10×reaction buffer,4mmol/L MgCl₂,250μmol/L dNTP ,2.5U Taq DNA polymerase and 0.4 μmol/L. The suitable PCR procedure was one cycle denaturing at 94 ℃ 5min,40 cycles which involves denaturing at 94 ℃ for 30 s, annealing at 36 ℃ for 30 s and extending at 72 ℃ for 1 min, one cycle extending at 72 ℃ for 7 min.4, 92 Repeatable loci (including 76 polymorphic loci) were amplified using 9 random primers in 42 watermelon resources. The percentage of polymorphic bands was 82.6%, 10.2 polymorphic bands per primer were amplified 5-16repeatable loci were amplified with each primer.5,As analyzed by NTSYSpc version 2.10, the genetic distance was between 0.0655-0.5081 .UPGMA clustering analysis based on genetic similarity showed that 42 materials could be classified into 8 groups which were one Russia group, one Japan group, one American group, one south of china group, one Northwest group and three medial groups. They were similar with those classified by traditional watermellon method.