柳杉无性系指纹图谱的构建及遗传多样性分析

Analysis of genetic diversity and construction of DNA fingerprinting of clones in Cryptomeria fortune

【目的】为了构建柳杉无性系的DNA指纹图谱,准确区分柳杉无性系。【方法】利用SSR标记分析了柳杉89个无性系的遗传多样性和指纹图谱。【结果】11对引物共检测出53个等位点,平均每个SSR位点有5个,变化范围为3~6个;有效等位基因数(Ne)变化范围为1.887 0~4.295 6,平均为3.041 1;Shannon信息指数(I)变化范围为0.774 9~1.556 3,平均为1.208 1。表明SSR标记具有较高的多态性。根据SSR标记特点及引物的顺序,将SSR引物扩增统计的"0"、"1"转换成基因型,通过不同基因型组合,构建了柳杉89个无性系的DNA分子指纹图谱,使每个无性系都获得了1个22位数的指纹图谱号码,同时基于个体间的遗传距离将柳杉89个无性系分为5类。【结论】SSR标记适用于柳杉无性系遗传多样性分析及鉴定,柳杉的遗传多态性处于中等偏高水平。

【Objective】Molecular fingerprinting and genetic diversity analysis are important for protecting and managing superior germplasm resources,as well as for identifying superior clones.【Method】In the present study,11 SSR primer pairs were used to assess the genetic diversity and DNA fingerprints of 89 C.fortunei clones.【Result】A total of 53 putative alleles were detected from 11 pairs of primers,with a mean of five putative alleles per locus,ranging from three to six.The number of effective alleles（Ne） ranged from 1.887 0 to 4.295 6,with a mean of 3.041 1,and the Shannon＇s information index ranged from 0.774 9 to 1.556 3,with a mean of 1.208 1.These findings indicate that SSR markers in C.fortunei exist a relative high polymorphism.Furthermore,according to the characteristics of the SSR markers and primer sequences,the present（recorded as 1） and absent（recorded as 0） collected from amplification by SSR markers were transferred into genetic types.By combining different genetic types code,we established DNA fingerprints of 89 C.fortunei clones,and attached a 22-digit fingerprint number to each clone.The 89 clones were also separated into five clusters based on genetic distance.【Conclusion】SSR markers are suitable for analyzing and identifying the genetic diversity of C.fortune,and C.fortune shows moderate to high genetic diversity.