CAJ | 학술논문

以提取的苜蓿基因组DNA为模板，通过正交设计，筛选出ISSR-PCR反应体系中最适宜的各组分浓度，即20μL的反应体系中最适添加量分别为2．0 U/μL的Taq DNA聚合酶，0．3 mmol/L的dNTP，1．0 mmol/L的MgCl2，0．1μmol/L的ISSR引物以及30 ng/μL的DNA模板。在此基础上对30份不同苜蓿种质材料进行遗传多样性研究，通过聚类分析研究，将30份不同苜蓿种质材料分为四大类，第一大类为来自加拿大的4个苜蓿品种（系）；第二大类为来自美国的12个苜蓿品种（系）；第三大类为来自中国和荷兰的12个苜蓿品种（系）；第四大类为来自澳大利亚的2个苜蓿品种（系）。
이제취적목숙기인조DNA위모판，통과정교설계，사선출ISSR-PCR반응체계중최괄의적각조분농도，즉20μL적반응체계중최괄첨가량분별위2．0 U/μL적Taq DNA취합매，0．3 mmol/L적dNTP，1．0 mmol/L적MgCl2，0．1μmol/L적ISSR인물이급30 ng/μL적DNA모판。재차기출상대30빈불동목숙충질재료진행유전다양성연구，통과취류분석연구，장30빈불동목숙충질재료분위사대류，제일대류위래자가나대적4개목숙품충（계）；제이대류위래자미국적12개목숙품충（계）；제삼대류위래자중국화하란적12개목숙품충（계）；제사대류위래자오대리아적2개목숙품충（계）。
Template DNA used for ISSR was extracted from alfalfa leaf tissue .The orthogonal design was ap-plied to optimize ISSR amplification system .Anoptimal reaction system was established ,20 μL reaction system con-tained 2.0 U/μL Taq DNA polymerase,0.3 mmol/L dNTP,1.0 mmol/L MgCl2,0.1 μmol/L ISSR primers and 30 ng/μL DNA template .Genetic diversity studies of Medicago L.germplasms were based on the result of the orthogo-nal design .And by cluster analysis ,30 varieties were divied into four groups , one group was consist of four alfalfa varieties coming from Canada , the second group was composed of twelve varieties coming from America , the third group includes twelve alfalfa varieties coming from China and Netherland ,and the last group has two alfalfa varieties coming from Australia .