The Application Of ISSR Markers On The Relationship Of Rice Cultivars And Cultivar Identification

In this study, fingerprints of 70 rice cultivars in Jilin province were established by ISSR markers. The results showed that:1. High quality DNA could be extracted from young rice seedlings by SDS method to meet the requirement of ISSR amplification.2. The optimum ISSR reaction system were determined as 0.5μl primer, 0.4 μl 2.5mM dNTPs, 2. 5μl 10× PCR buffer, 2U Taq DNA polymerase. The PCR mixture were denatured at 94℃ for 4 min, Cycling consisted of 30 sec at 94℃, 60 sec at 58℃, and 90 sec at 72℃ for 40 cycles and then followed by elongation at 72℃ for 10 min on a Biometra T Gradient DNA Thermal Cycler.3. 27 ISSR primers were screened out from total 130 primers for they showed polymorphic bands based on the amplification results of 3 cultivars. The fingerprints of 70 rice cultivars in Jilin province were established by 20 ISSR markers.The clustering results of 70 rice cultivars showed that the cultivars with relationship tended to be get together and was similar to that of pedigree analysis. The results also indicated that ISSR markers could be used in the study of rice relationship and rice cultivar identification.