华北农学报 ›› 2009, Vol. 24 ›› Issue (5): 107-112. doi: 10.7668/hbnxb.2009.05.023

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新麦草种质遗传多样性的ISSR分析

刘永财1,2, 孟林1, 张国芳1, 毛培春1, 张德罡2   

  1. 1. 北京市农林科学院北京草业与环境研究发展中心, 北京 100097;
    2. 甘肃农业大学草业学院, 甘肃兰州 730070
  • 收稿日期:2008-12-23 出版日期:2009-10-28
  • 通讯作者: 孟林(1966-),男,内蒙古乌兰察布人,副研究员,博士,主要从事草业科学研究。
  • 作者简介:刘永财(1981-),男,吉林松原人,硕士,主要从事牧草种质资源研究。
  • 基金资助:
    北京市科技计划项目(209090501040902);农业部牧草种质资源保护项目

Genetic Diversity Analysis of 15 Psathyrostachys juncea Germplasm Resources by ISSR Molecular Marker

LIU Yong-cai1,2, MENG Lin1, ZHANG Guo-fang1, MAO Pei-chun1, ZHANG De-gang2   

  1. 1. Beijing Research and Development Center for Grasses and Environment, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;
    2. Pratacultural College, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2008-12-23 Published:2009-10-28

摘要: 利用ISSR标记对15份新麦草(Psathyrostachys juncea)种质资源进行遗传多样性检测。为获得稳定的新麦草ISSR反应体系,对反应条件中的dNTP、Taq酶浓度、引物浓度、Mg2+浓度等因子进行了优化,建立起的最佳反应体系为:25μL的反应体系中含有dNTP 2.5μL、Taq酶0.2μL、Primers 1μL、Mg2+1.5μL、2.5μL 10×Buffer、2μL模板DNA和ddH2O 15.3μL。ISSR标记结果显示,筛选出的重复性好、谱带清晰的ISSR引物8条,共获得84个扩增位点,其中多态性位点72个,多态性比率(PPB)为85.7%。通过遗传相似系数(GS)的计算,供试种质材料间的GS值在0.4405~0.9048之间,表现出丰富的遗传多样性。通过聚类分析,将15份新麦草种质材料划分为四大类,且呈现出一定的地域性分布规律。

关键词: 新麦草, ISSR分子标记, 遗传多样性, 聚类分析

Abstract: The genetic diversities of 15 Psathyrostachys juncea germplasm resources were analyzed by ISSR molecular marker.Based on the basic system,four factors of Taq DNA polymerase,random primers,Mg2+,and dNTPs which affect ISSR-PCR amplifying results of P.juncea were optimized.An optimal ISSR-PCR system including 2.5μL dNTPs,2μL template DNA,0.2μL Taq DNA polymerase,1μL random primers,1.5μL Mg2+,2.5μL 10×PCR Buffer,15.3μL ddH2O,and the total volume was 25μL,was established.The results of ISSR marker showed that 8 primers,which could show clearer bands and had good repetition,were selected from 25 primers and detected a total of 84 bands,among which 72 bands were polymorphic.The average percentage of polymorphic bands (PPB) was 85.7%.According to 0.4405-0.9048 of the GS values,the germplasm and materials of P.juncea showed rich genetic diversity.Based on cluster analysis of the genetic characteristics,15 P.juncea germplasm could be divided into 4 groups according to the nearest phylogenetic relationship.In most cases,the germplasm and materials from the same country were in the same group and showed the definite rules of geographical distribution.

Key words: Psathyrostachys juncea, ISSR molecular marker, Genetic diversity, Cluster analysis

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引用本文

刘永财, 孟林, 张国芳, 毛培春, 张德罡. 新麦草种质遗传多样性的ISSR分析[J]. 华北农学报, 2009, 24(5): 107-112. doi: 10.7668/hbnxb.2009.05.023.

LIU Yong-cai, MENG Lin, ZHANG Guo-fang, MAO Pei-chun, ZHANG De-gang. Genetic Diversity Analysis of 15 Psathyrostachys juncea Germplasm Resources by ISSR Molecular Marker[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(5): 107-112. doi: 10.7668/hbnxb.2009.05.023.

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