CAJ | 학술논문

采用烟草靶斑病菌YC-9, LJT-8和QYS-7为DNA模板，初步筛选SRAP引物组合；采用L16(45)正交试验设计，对烟草靶斑病菌的SRAP-PCR反应体系中的Mg2+、dNTPs, Taq DNA聚合酶、引物和DNA模板浓度等5个因素进行优化试验。结果表明：共筛出13对扩增条带清晰且多态性好的引物组合；烟草靶斑病菌的最佳SRAP反应体系为Mg2+浓度2.0 mmol/L、dNTP浓度200μmol/L、Taq DNA聚合酶0.8 U、引物浓度140 mmol/L、模板DNA 20 ng及1×PCR buffer，反应总体积为20μL；各因素对SRAP-PCR扩增反应结果影响的差异较大，依次为Taq DNA聚合酶>引物> Mg2+> dNTPs=模板DNA。
채용연초파반병균YC-9, LJT-8화QYS-7위DNA모판，초보사선SRAP인물조합；채용L16(45)정교시험설계，대연초파반병균적SRAP-PCR반응체계중적Mg2+、dNTPs, Taq DNA취합매、인물화DNA모판농도등5개인소진행우화시험。결과표명：공사출13대확증조대청석차다태성호적인물조합；연초파반병균적최가SRAP반응체계위Mg2+농도2.0 mmol/L、dNTP농도200μmol/L、Taq DNA취합매0.8 U、인물농도140 mmol/L、모판DNA 20 ng급1×PCR buffer，반응총체적위20μL；각인소대SRAP-PCR확증반응결과영향적차이교대，의차위Taq DNA취합매>인물> Mg2+> dNTPs=모판DNA。
SRAP primer pairs were screened for polymorphism using DNA of Rhizoctonia solaniisolates YC-9, LJT-8 and QYS-7 as templates. An orthogonal design of L16(45) was used to optimize SRAP-PCR reaction system forR. solaniof tobacco with 5 factors, namely Mg2+, dNTPs, primers, Taq DNA polymerase and template DNA. Results showed that a total of 13 polymorphic SRAP primer pairs were screened out of 100 SRAP primer pairs, and a suitable SRAP-PCR reaction system for Rhizoctonia solanifrom tobacco target spot was 2.0 mmol.L-1 Mg2+, 200 mmol.L-1 dNTPs, 0.8U Taq DNA polymerase, 140 mmol.L-1 primer pairs, 20 ng template DNA and 1×PCR buffer. In addition, each factor in SRAP-PCR reaction system had different effects on amplified patterns in descending order of Taq DNA polymerase> primer>Mg2+> dNTPs=DNA.