摘要
建立和优化了花生疮痂病菌ISSR-PCR反应体系,为研究花生疮痂病菌遗传多样性及遗传结构提供理论基础。采用正交试验和细调性单因素试验对模板DNA用量、最适引物、引物浓度、Mg^(2+)浓度、Taq酶浓度、d NTPs浓度以及退火温度等影响ISSR反应结果的各因素进行了研究。最优体系中各成分浓度分别为Mg^(2+) 2. 0 mmol/L,dNTPs 0. 2 mmol/L,Taq酶1. 5 U,引物0. 88μmol/L,模板DNA 30～40 ng/25μL;筛选出8条适用于花生疮痂病菌遗传多样性分析的ISSR引物。
In order to provide a basis for the research of genetic diversity and genetic structure of peanut scab,ISSR-PCR reaction system of Elsino■arachidis was established and optimized.The factors influencing ISSR-PCR reaction results,including template DNA dosage,optimum primer,primer concentration,Mg2+concentration,Taq enzyme concentration,dNTPs concentration and annealing temperature,were studied by fine-tuning single factor test and orthogonal test.The concentration of each component in the optimal system was as follows:2.0 mmol/L Mg2+,0.2 mmol/L dNTPs,1.5 U Taq DNA polymerase,0.88μmol/L primer,30-40 ng template DNA.Eight ISSR primers were screened for genetic diversity analysis of Elsino■arachidis.
作者
李元杰
刘璐
周如军
赵杰锋
付祎玮
傅俊范
LI Yuanjie;LIU Lu;ZHOU Rujun;ZHAO Jiefeng;FU Yiwei;FU Junfan(College of Plant Protection,Shenyang Agricultural University,Shenyang 110866,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2019年第2期142-148,共7页
Journal of Jilin Agricultural University
基金
辽宁省农业领域青年科技创新人才培养计划项目(2014056)
