SSR Research On Genetic Diversity Of Germplasm Resources Of Lilium Distributed In The Middle Reach Of Yangtze River

Since there are the rich wild lily germplasm resources in China, including55native ones in115lily speceies, it has a great significance to study the genetic diversity of Chinese Lilium. In recent years, more and more studies on it have been made, however, studies which choose the wild lily germplasm resources from the middle reaches of Yangtze River is really rare. In this study, we have used SSR marker to analyze the genetic diversity of the253samples of10Lillium species collected from the middle reaches of the Yangtze River, hoping to provide some help for the germplasm utilization and conservation in this region.The main results are as follows:1. The establishment of the lily SSR test procedureA set of SSR electrophoresis procedure of Lilium based on denaturing polyacrylamide gel was founded, at the same time, we succeed to choose the optimizing SSR-PCR system. The study has found that:the quality of DNA template has litter effect on the amplification; the tough-down PCR procedure is not obviously better than the ordinary PCR procedure; the bands are clear when the sample volume in2.5μl/hole to4μl/hole; the separation effect of denaturing polyacrylamide gel electrophoresis is better than the non-denaturing; the optimizing SSR-PCR system:Mg2+3.0mmol.L-1, DNA100ng, Taq1.0U.μl-1, dNTP0.2mmol.L-1, adding pure water to20μl.2. The genetic diversity analysis of the wild lily germplasm resources from the middle reaches of the Yangtze River.There are6pairs of SSR primers screened,42alleles detected, each pair of primer has amplified7alleles on average.The polymorphic percentage is100%. The genetic diversity sequence from high to low is L. brownii, L. brownie var. viridulum, L. concolor, L. lancifolium, L. davidii, L. leucanthum, L. henryi, L. distichum, L. rosthornii, L. speciosum.The observed heterozygosity among10lily species is volatile, mean of0.4877, the heterozygosity is high. Genetic differentiation has appeared between specieses. Only the heterozygosity of L. concolor is excess. Overall, the genotype is very rich. The genetic variation within populations is less than it between populations, the gene flow Nm is less than1, and the genetic differentiation of Lilium from the Yangtze River middle reaches is serious.Clustering analysis showed10lily specieses was divided into three groups, the first group is L. henryi, L. rosthornii, L. speciosum; the second group is L. concolor, L. lancifolium; the remaining5species together into the third group.3. The genetic diversity analysis of L. browniiThe mean of the number of alleles (A), the effective number of alleles (Ae), the Shannon information index (I) of5L. brownii populations respectively are3.7667,2.7758,1.0607. The genetic diversity sequence is WS>SNJ>BD>ZG>YC.L. brownii has a low heterozygosity, the heterozygosity is deficient.82genotypes have been found among91samples, the genotype is very rich. The genotypic diversity (k) is from0.8000to0.9630, SNJ is the highest.The genetic variation of the L.brownii within the population is higher than it among the population. The gene exchange exists between populations. Clustering analysis showed5populations have been divided into2groups, YC alone into one group, the rest4populations into another.4. The genetic diversity analysis of L. leucanthumThe mean of the number of alleles (A), the effective number of alleles (Ae), the Shannon information index (Ⅰ) of4L. leucanthum populations respectively are2.7084,1.7699,0.6489. The genetic diversity sequence is DSN>BJP>CBD>AJP.L. leucanthum has a low heterozygosity, and CBD is the highest population, and heterozygosity is deficient.40genotypes have been found among44samples, the genotype is rich. The genotypic diversity (k) is from0.8000to1.0000, CBD and DSN is the highest, while AJP is the lowest.The genetic variation of the L. leucanthum within the population is higher than it among the population. The mean of Nm is1.1116, and gene flow exists between populations. Clustering analysis showed4populations have been divided into2groups, DSN alone into one group; the genetic distance between AJP and BJP is the closest, first together into a small branch, then clustered with CBD into another group.