渔业科学进展2012，Vol.33Issue(4)：72-82,11.
仿刺参微卫星标记的筛选及群体遗传结构分析
Development of microsatellite DNA markers of sea cucumber Apostichopus japonicus and their application in population genetic variation and structure analysis
摘要
Abstract
In order to evaluate the genetic diversity of sea cucumber Apostichopus japoni- cus and breed eugenic varieties, a CA microsatellite containing short fragment genomic DNA library of sea cucumber was constructed using the FIASCO (Fast Isolation by AFLP Sequences Containing Repeats)method. In 140 randomly selected and sequenced clones, 118 contained a CA repeat motif. According to Weber(1990) classification rules, the sequences were divided into three categories; 83 perfect repeat sequences without interruptions in the runs of CA or GT dinucleotides (70.4% of total) , 30 imperfect repeat sequences with one or more interruptions in the run of repeats (25. 4%) , and 5 compound repeat sequences with adjacent tandem simple repeats of a different sequence (4. 2%). Twenty pairs of primers were designed and used to investigate the polymorphism and genetic structure of wild A. japonicus collected from China(CH) , Korea(KW, KE) , Japan(JA) and Russia(RU). Cross amplification of twenty loci were also tested in Parastichopus parvimensis which were collected in USA (AM). The results demonstrated that all the markers were polymorphic and 16 loci had high polymorphism (PIC>0. 5). The average observed and expected heterozygosities of the 20 loci were 0. 39 and 0. 69, respectively. In addition, a total of 231 alleles (102 effective alleles) were detected at 20 loci in the total samples and the alleles number of each locus ranges from 3 to 20 with average of 11. 6. The Hardy-Weinberg Equilibrium (HWE) analysis showed that all the loci deviated from genetic e-quilibrium in the six populations and heterozygote deficiency occurred to different extent in total samples (Fis>0). Genetic identity of the five A. japonicus populations was 0.71, while the genetic identity between A. japonicus and P. parvimensis were quiet low. In the UPMGA tree, CH and KW populations formed a cluster, RU, JA and KE populations formed another cluster, while AM felt into a seperate cluster. The result of cluster analysis indicated that the clustering order had significant correlation with geographic distance.关键词
刺参/微卫星标记/FIASCO/遗传多样性/遗传结构Key words
Apostichopus japonicus/Microsatellite DNA markers/FIASCO Genetic diversity/Genetic structure分类
生物科学引用本文复制引用
潘传燕,臧云鹏,廖梅杰,王印庚,荣小军,张正,李彬,陈贵平..仿刺参微卫星标记的筛选及群体遗传结构分析[J].渔业科学进展,2012,33(4):72-82,11.基金项目
国家自然科学基金项目(刺参弧菌抗性相关基因筛选与表达研究)、国家高技术研究发展计划项目(2012AA10A412)、山东省农业良种工程重大课题“速生、抗病、耐高温刺参良种选育”、科研院所技术开发研究专项(2011EG34219)和中央级公益性科研院所基本科研业务费专项资金项目(20603022012006)共同资助 （刺参弧菌抗性相关基因筛选与表达研究）
