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| 苦荞SRAP分子标记体系优化与遗传多样性分析 |
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令狐斌1, 侯思宇1,2,3, 孙朝霞1, 黄可盛1, 路阳1, 韩渊怀1,2,3, 许冬梅1
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| (1.山西农业大学 农学院, 山西 太谷 030801;2.山西农业大学 农业生物工程研究所, 山西 太谷 030801;3.山西省农业科学院 农业部黄土高原作物基因资源与种质创制重点实验室, 太原 030031) |
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| 摘要: |
| 通过正交试验设计,比较苦荞序列相关多态性-聚合酶链式反应(Sequence related amplified polymorphism-polymerase chain reaction,SRAP-PCR)扩增反应体系的组合,选出最佳扩增体系用于苦荞SRAP分子标记进行遗传多样性分析。其中最佳SRAP-PCR反应扩增体系为Taq DNA聚合酶2 U、Mg2+ 1.5 mmol/L、DNA模板75 ng、引物0.5 μmol/L。从238对SRAP引物组合中筛选出20对多态性较高的引物组合,并对15个苦荞品种进行遗传多样性分析。结果表明:20对引物组合共扩增出128个位点,其多态性信息量(Polymorphism information content,PIC)值在0.66~0.95,每对引物组合扩增位点为7~13个,多态性比率平均值为81.6%,其中多态性信息量值为0.95的引物组合为me7em11、me9em4和me12em8。基于UPGMA法聚类(GS=0.78)可将15个苦荞品种划分为3大类,但这3类划分的品种没有明显的地域分布特征。 |
| 关键词: 苦荞 SRAP-PCR 正交试验设计 遗传多样性 |
| DOI：10.11841/j.issn.1007-4333.2015.01.005 |
| 投稿时间：2014-05-06 |
| 基金项目:国家自然科学基金(NSFC:31301385); 山西省青年科技研究基金(2011021032-1,2011021032-3); 山西农业大学科技创新育种基金(2010028); 高等学校科技创新项目(2013115); 山西省高校重点学科建设专项资金 |
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| Optimization of SRAP-PCR amplification system and its application in the analysis of genetic diversity in Tartary buckwheat |
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LINGHU Bin1, HOU Si-yu1,2,3, SUN Zhao-xia1, HUANG Ke-sheng1, LU Yang1, HAN Yuan-huai1,2,3, XU Dong-mei1
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| (1.College of Agriculture, Shanxi Agricultural University, Taigu 030801, China;2.Institute of Agricultural Biotechnology, Shanxi Agricultural University, Taigu 030801, China;3.Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau of Ministry of Agriculture, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China) |
| Abstract: |
| Through orthogonal experimental design,the optimum SRAP-PCR(Sequence related amplified polymorphism-polymerase chain reaction) amplification system was developed to estimate the genetic diversity by comparing the effect of four main PCR reaction components.The best SRAP-PCR amplification system comprised of 2 U Taq DNA polymerase,1.5 mmol/L Mg2+,75 ng DNA template,0.5 μmol/L primer.Twenty pairs of primers from the 238 pairs of primer combinations were selected by PCR product polymorphism among 15 Tartary buckwheat cultivars.The results showed that 128 loci were amplified by 20 pairs of primer combinations.The polymorphism information content (PIC) values of these primer combinations were between 0.66 to 0.95.The amplified loci for each pair of primer combination were 7-13,and polymorphism average ratio was 81.6%.The information value for primer combination of me7em11,me9em4 and me12em8 was 0.95.The 15 Tartary buckwheat cultivars could be divided into three categories based on UPGMA (GS=0.78),but no obvious geographical distribution characteristics were observed.We optimized SRAP-PCR molecular marker system to analyze genetic diversity among 15 Tartary buckwheat cultivars.It laid the theoretical foundation for research on the germplasm resources of Tartary buckwheat. |
| Key words: Tartary buckwheat SRAP-PCR orthogonal experimental design genetic diversity |
