RT-PCR检测甜菜坏死黄脉病毒及总RNA提取方法研究  

The Studies on RT-PCR Detection Method of Beet Necrotic Yellow Vein Virus(BNYVV) and the Extraction Method of Total RNA

作　　者：牛素清[1] 白晨[1] 张惠忠[1] 李晓东[1] 付增娟[1] 赵尚敏[1] 斯琴巴特尔[2] 轩继雨[1] 李树生[1] 

机构地区：[1]内蒙古农牧业科学院甜菜研究所,内蒙古呼和浩特010031 [2]内蒙古农牧业科学院生物中心,内蒙古呼和浩特010031

出　　处：《华北农学报》2008年第1期207-210,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家“863”计划资助项目(2001AA241193)

摘　　要：为选择较为适合甜菜总RNA的提取方法,构建和优化RT有效体系。以田间甜菜叶片、根毛及根表皮为材料,研究了提取甜菜总RNA的方法以及利用RT-PCR技术进行甜菜坏死黄脉病毒(BNYVV)的检测。研究表明,获得良好的RT-PCR扩增效果,RT体系中dNTPs浓度、引物、AMV、模板RNA的浓度要分别达到1 mmol/L,1μmol/L,0.1 U/μL,0.01μg/μL较好。PCR体系中dNTPs浓度、引物、TaqDNA聚合酶、Mg2+的浓度要分别达到0.1 mmol/L,0.1μmol/L,0.01U/μL,1.48 mmol/L效果较好。RT-PCR法对BNYVV检测,可作为甜菜品种(育种材料)早期抗丛根病性鉴定的依据之一。Two methods of extraction of total RNA from sugarbeet leaves, cortexes and root hair were discussed. Optimized conditions for RT-PCR method for detecting BNYVV were also studied in this paper. The results were as follow, for getting the effective RT-PCR detection system, the better Concentration of RT reaction ingredients dNTPs, primer, AMV and template RNA were 1 mmol/L, 1 μmol/L, 0.1 U/μL, 0.01 μg/μL respectively. The concentration of ingredients dNTPs,primer,Taq DNA polymerase and Mg^2＋ should achieve 0.1 mmol/L,0.1 μmol/L,0.01 U/μL, 1.48 mmol/ L in PCR reaction system.

关 键 词：甜菜 甜菜坏死黄脉病毒 RT-PCR 

分 类 号：S566.3[农业科学—作物学]