Development Of SSR Markers And The Study Of Genetic Diversity Of Tapiseia Sinensis Oliv.

Tapiseia sinensis Oliv is a deciduous tree that belongs to Staphyleaceae family. The study of it has scientific value to the division of Chinese subtropical flora and the systematic development of Staphyleaceae family. The leaves of Tapiseia sinensis Oliv are rich in flavonoids compounds and have significant biological activities of antiulcer, anti-inflammatory, and reducing blood fat, and are thus widely used in medicine. In addition, Tapiseia sinensis Oliv has soft wood material, straight texture, and smooth surface, and thus has high practical value of making furniture, sheet, etc. At the same time, the shape of the tree is beautiful with tall and straight trunk, and yellow leaves in autumn, and can be also used for garden greening.This study is based on a field investigation and group sampling of the rare and endangered plant Tapiseia sinensis Oliv. It makes a specific SSR primer development through magnetic beads enrichment and dual-suppression-PCR technique, and makes an analysis of the genetic diversity of two populations by using the screening of high polymorphic primers. Specific research results are as follows:1. Through magnetic beads enrichment420positive clones were screened out. Through sequencing70pairs of SSR primers were synthesized successfully. By PCR amplification screening, finally13pairs of primers were obtained to have band amplification,5pairs had polymorphism amplification, and8pairs were monomorphism primers, and polymorphic SSR primers occupied only7.15%.220positive clones were screened out through dual-suppression-PCR technique. Through sequencing41pairs of SSR primers were synthesized successfully. By PCR amplification screening, finally14pairs of primers were developed to have band amplification,6pairs had polymorphism amplification, and8pairs were monomorphism primers, and polymorphic SSR primers occupied14.63%.2. Using the Tapiseia sinensis Oliv Pop of the66samples in Shunhuang Mountain in Hunan and Wuyi Mountain in Jiangxi, this study made an analysis of testing11pairs of SSR primers with polymorphism amplification got through the above two methods,60allele bands were amplified, among which42were polymorphic bands and occupied70%. Calculating its genetic diversity parameters with GenALEx software, The number of alleles was2-9, average allele number was3.818. The observed heterozygosity was0.017-0.602, and the mean observed heterozygosity was0.317; the expected heterozygosity was0.208-0.543, and the average expected heterozygosity was0.378. All SSR loci did not deviate from the Hardy-Weinberg equilibrium, and there was no linkage disequilibrium phenomenon.in the SSR loci of different populations. So,11pairs of SSR primers with polymorphism amplification got through this study were the unique to Tapiseia sinensis Oliv.3. The results of genetic analysis were that in population of Hunan Shunhuang Mountain, the number of alleles was1-3, mean number of allele was1.909, the number of effective allele was1-2.352, and the mean number was1.460. The observed heterozygosity was0～0.621, and the mean observed heterozygosity was0.160; the expected heterozygosity was0～0.575, and the average expected heterozygosity was0.390, and this results showed that genetic diversity of Hunan Shunhuang Mountain was low, and genetic differentiation was small. In population of Jiangxi Wuyi Mountain, the number of alleles was1～8, mean number of allele was3.545, the number of effective allele was1-3.442, and the mean number was2.245. The observed heterozygosity was0～0.750, and the mean observed heterozygosity was0.475; the expected heterozygosity was0～0.709, and the mean expected heterozygosity was0.508, and this results showed that there existed a middle level of genetic diversity in Jiangxi Wuyi Mountain. GenALEx software was used to analyze its genetic structure, and the results showed that there existed a clear genetic differentiation (FST=0.379) in the same population, between Hunan Shunhuang Mountain and Jiangxi Wuyi Mountain, the genetic flow was relatively low (Nm=0.521), and there also existed relatively obvious genetic differentiation.