摘要
白背飞虱是水稻重要害虫之一.研究采用L16 （45）正交设计优化试验,构建了白背飞虱ISSR最优反应体系,并对引物退火温度和反应循环次数进行了优化.结果表明：白背飞虱ISSR-PCR最优反应体系为2μL 10×PCRBuffer（10 mmol/L Tris-HCl; 50 mmol/L KCl）,1.5 mmol/L Mg2＋,0.25 mmol/L dNTPs,0.9μmol/L引物,1U TaqPolymerase和50 ng模板DNA,ddH2O补充至20μL.本研究采用的10条引物的最佳退火温度在51.0～54.0℃间,以引物相应最优退火温度和40次反应循环可获得较好的白背飞虱ISSR-PCR结果.
Sogatella furcifera is one of the most important rice pests.To establish and optimize ISSR-PCR reaction system for S.furcifera,the factors influencing ISSR system were studied with L16 （45） orthogonal design,and the optimal annealing temperature and cycle times were proposed.The results showcd that the optimal conditions for ISSR-PCR system were 2μL 10×PCR buffer （10 mmol/L Tris-HC1,50 mmol/L KCl）,1.5 mmol/L Mg2＋,0.25 mmol/L dNTPs mixture,0.9 μmol/L of each primer,1U Taq DNA polymerase （TaKaRa） and 50 ng template DNA in 20μL reaction volume.The optimal annealing temperatures were 51.0-54.0 ℃ for the 10 primers used in the present study.The clear and reproducible DNA bands were obtained by the optimal annealing temperature and 40 reaction cycles.
出处
《植物保护》
CAS
CSCD
北大核心
2014年第2期109-112,共4页
Plant Protection
基金
贵州省农业攻关(黔科合NY字[2010]3064号)
"973"计划前期研究专项(2009CB125909)
贵州省教育厅自然科学研究项目(2010011)
关键词
白背飞虱
正交设计
ISSR
Sogatella furcifera
orthogonal design
ISSR
