Genetic Diversity Of Chimonanthus Praecox (L.) Link Based On ISSR Markers

Chimonanthus praecox (L.) Link,the special plant of china,is one of the famous and precious flower plants which have long cultural history.It has a broad distributing and many cultivars.In this study,29 cuitivars were used to analyse genetic diversity and phylogentic relationship by ISSR and morphological markers.The main results were presented as following:1.Spend part of phenotype diversity 11 characters including principal component analysis to 29 different chimonanthus praecox (L.) Link materials, it is concluded that the "pattern", "colour of inside bloom ", "flower diameter", "bloom color" have important effects on cultivars classincation."colour of inside bloom"should be regarded as the first standard, three other features can be used as the secondary variety classification retrieval standards.2.An optimized PCR reaction system for ISSR was established,optimum ISSR reaction system of Chimonanthus praecox was every 20μl reaction system containing 10×PCR buffer,1.6μl Mg2+ (25mmol/L).0.6μl dNTPs(dATP,dCTP,dGTP,dTTP each 10mmol/L)2.0μl primer,1.5 U Taq DNA polymerase,30ng DNA template.The amplication was 1 cycle after initial denaturation at 94℃for 5 min,follwed by 35 cycle of 1 min at 94℃,l min annealing at 52～59℃(depending on different primers),1 min 1 0 seconds at 72℃,and a final l 0 min extension at 72℃,After which the samples were held at 4℃.3.Ten primers with good,clear bands were got,screening from one hundred ISSR primers.These ten primers were used to amplicated respectively the genomic DNA of 29 different chimonanthus praecox (L.) Link materials,.And a tatal of 75bands were amplined with 38 polymorphic bands and precentage of polymorphic bands(PPB)50.7%.The average value of observed number of alleles average value of observed number of alleles,Nei's gene diversity and Shannon,s information index were 1.5325,1.2956,0.1873,0.2321 respectively.