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。

关键词: 甜菜, 甜菜坏死黄脉病毒, RT-PCR

Abstract: Two methods of extraction of total RNA from sugarbeet leaves,cortexes and root hairwere discussed. Opti-mized conditions for RT-PCRmethod for detecting BNYVV were also studied in this paper. The resultswere as follow,forgetting the effective RT- PCR detection system, the better concentration of RT reaction ingredients dNTPs, primer,AMVand template RNA were 1 mmol/L, 1 μ mol/L, 0. 1 U/μ L, 0. 01 μ g/μ L respectively. The concentration of ingredientsdNTPs,primer,Taq DNA polymerase and Mg2 +should achieve 0. 1 mmol/L,0. 1 μ mol/L,0. 01 U/μ L,1. 48mmol/L inPCR reaction system.

Key words: Sugarbeet, Bat Necvotic Yellow Vein Virus, RT-PCR

中图分类号: 

• S566.3