甘蓝型油菜SSR检测体系的优化  

Optimization of SSR Detection System in Rapeseed(Brassica napus L.)

作　　者：张洁夫[1,2] 戚存扣[3] 浦惠明[3] 陈锋[3] 顾慧[3] 陈新军[3] 高建芹[3] 傅寿仲[3] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室 [2]江苏省农业科学院经济作物研究所,江苏南京210014 [3]江苏省农业科学院经济作物研究所

出　　处：《江苏农业学报》2004年第4期225-229,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目 ( 3 0 170 5 83 ) ;江苏省高技术项目(BG2 0 0 2 3 0 3 )

摘　　要：油菜SSR检测是其主要农艺性状分子标记及标记辅助育种的一项重要技术。以无花瓣品系APL0 1、常规有花瓣抗菌核病品种M 0 83及其回交群体BC1为供试材料 ,研究油菜SSR检测体系中PCR扩增的各主要成分对检测结果的影响 ,比较不同电泳方法及染色方法的效果 ,进一步优化了适用于油菜的SSR检测体系。优化后的PCR反应体系为 :1×Buffer(含 2 0 0mmol/LMgCl2 )、5 0 0 0 μmol/LdNTPs、0 5 0UTaq酶、0 0 8μmol/LSSR引物、6 0 0ng/μl模板DNA ,加ddH2 O至 10 0 0 μl。SSR detection is an important technique of molecular marker for main agronomical traits and marker-(assisted) breeding in rape seed.An apetalous rape seed line APL01,a conventional petalous variety M083 and its backcross population were used as test materials.Effect of major component of PCR amplification on results detected in SSR detection system was studied,different electrophoresis and staining methods were compared,and SSR detection system suitable for rapeseed was established and optimized.The optimized PCR amplification system was 1×Buffer (including 2.00 mmol/L MgCl_2),50.00 μmol/L dNTPs,0.50 U Taq enzyme,0.08 μmol/L SSR primer,6.00 ng/μl template DNA,add ddH_2O to terminal volume 10.00 μl.The effect of polyacrylamide gel electrophoresis was better than that of agarose gel electrophoresis,and silver staining was better than EB staining.

关 键 词：甘蓝型油菜 SSR 检测体系 优化 

分 类 号：S565.4[农业科学—作物学]