CAJ | 학술논문

采用正交试验对烟草抗PVYN突变株SN01幼嫩叶片的SRAP-PCR反应体系中5个主要因素（dNTPs、Mg2+、Taq DNA聚合酶、引物和DNA）进行了优化以建立合适的SRAP反应体系。结果表明，在20μL的反应体系中，各因素的最佳加入量分别是：dNTPs（2.5 mmol/L）3.2μL、Mg2+（25 mmol/L）1.6μL、Taq DNA聚合酶（2.5 U/μL）0.16μL、引物（10μmol/L）0.6μL、DNA模板120 ng。对该SRAP-PCR反应程序中的退火温度和循环次数筛选表明，最佳退火温度为57℃，循环次数为35次。该优化体系的建立，为进一步对烟草抗PVYN突变株SN01的抗病基因研究提供了可靠基础。
채용정교시험대연초항PVYN돌변주SN01유눈협편적SRAP-PCR반응체계중5개주요인소（dNTPs、Mg2+、Taq DNA취합매、인물화DNA）진행료우화이건립합괄적SRAP반응체계。결과표명，재20μL적반응체계중，각인소적최가가입량분별시：dNTPs（2.5 mmol/L）3.2μL、Mg2+（25 mmol/L）1.6μL、Taq DNA취합매（2.5 U/μL）0.16μL、인물（10μmol/L）0.6μL、DNA모판120 ng。대해SRAP-PCR반응정서중적퇴화온도화순배차수사선표명，최가퇴화온도위57℃，순배차수위35차。해우화체계적건립，위진일보대연초항PVYN돌변주SN01적항병기인연구제공료가고기출。
In order to establish a suitable SRAP reaction system, the orthogonal experiment was used to establish and optimize the five main factors (dNTPs, Mg2+, Taq DNA polymerase, primer, and DNA template) of sequence-related amplified polymorphism (SRAP) system with young leaves of tobacco mutant SN01 resistant to Potato virus Y-vein necrosis strain (PVYN). The results showed that the best amounts of factors in 20μL of the reaction system were:dNTPs (2.5 mmol/L) 3.2μL, Mg2+(25 mmol/L) 1.6μL, Taq DNA polymerase (2.5 U/μL) 0.16 μL, primer (10μmol/L) 0.6 μL, DNA template 120 ng, respectively. The best annealing temperature of 57℃and cycle times for 35 of the SRAP program were also conducted. This optimized system provides reliable basis for further study on the resistance genes of tobacco resistance mutant SN01.