[Objective]In order to identify the genetic differences among persimmon(Diospyros kaki)cultivars,a simple and efficient SNP molecular marker based on nrDNA internally transcribed spacer sequence was developed,which may provide reference for the collection,utilization and promotion of germplasm.[Method]The leaves of 18 hexaploid cultivated persimmon cultivars were used as experimental materials,and the ITS regions of 18 cultivars were amplified and sequenced,and sequence differences were analyzed.Finally,Sau96 I restriction endonuclides were used to verify 151 heterozygous sites.[Result]Sequencing results showed that ITS length of 18 persimmon varieties was 730 bp.Six heterozygous loci were found in the sequencing peak map,which were 151,168,205,278,279 and 622 bp.The proportion of base peak area of hexaploid persimmon heterozygous sites had special pattern,that is,C∶T= 2∶1,C∶T= 1∶1,C∶T= 1∶2 and A∶G= 1∶1.The 18 cultivated persimmon varieties could be divided into 11 categories according to the differences of heterozygous sites and the proportion of peak area.[Conclusion]The results of enzyme digestion at 151 sites verified that the concentration of enzyme digestion products was consistent with the proportion of its peak map area.The results showed that the SNP molecular labeling method based on nrDNA ITS sequence could divide 18 cultivated persimmon varieties into 11 groups.
维普
万方数据