RT-PCR方法同步检测兰州百合(Lilium davidii var.unicolor)两种主要病毒  

Simultaneously Detection of Two Main Lanzhou Lily (Lilium davidii var. unicolor) Viruses by RT-PCR

作　　者：张玉宝[1] 谢忠奎[1] 王亚军[1] 郭志鸿[1] 童勋章[1] 

机构地区：[1]中国科学院寒区旱区环境与工程研究所皋兰生态与农业综合试验站,兰州730000

出　　处：《武汉植物学研究》2010年第6期744-749,共6页Journal of Wuhan Botanical Research

基　　金：中国科学院农业项目办公室项目(CASN-115-06-08);中国科学院科技支甘工程专项(KJZG-2008-3)资助

摘　　要：建立并优化了以18S rRNA为内参照的特异性检测兰州百合(Lilium davidii var.unicolor)两种主要病毒:黄瓜花叶病毒(Cucumber mosaic virus,CMV)和百合无症病毒(Lily symptomless virus,LSV)的三重RT-PCR体系。结果表明,52.5°C的退火温度、0.025 U/μL的Taq DNA聚合酶、0.6 mmol/L的dNTP浓度、4 mmol/L的Mg2+浓度、0.4μmol/L的各引物浓度以及30个循环等是三重PCR体系扩增的最佳条件;同时用该优化体系检测了兰州百合不同取样部位的病毒差异,发现LSV在不同取样部位的特异扩增条带的强度比较一致,而CMV差距相对较大,外鳞片CMV相对含量在整个感病植株中最高。为兰州百合主要病毒检测、脱毒组培快繁提供了技术支撑。In this research,triplex PCR systems were established and optimized to specifically detect Cucumber mosaic virus（CMV） and Lily symptomless virus（LSV） of two main Lanzhou lily viruses with 18S ribosomal RNA used as an internal control.Results showed that an Annealing temperature of 52.5°C,Taq DNA polymerase of 0.025 U/μL,dNTP of 0.6 mmol/L,Mg2＋ of 4 mmol/L,three primer pairs of 0.4 μmol/L,and 30 cycles were the optimal amplification conditions for triplex PCR systems.Differences in viruses from different parts of Lanzhou lily were detected by the optimal triplex PCR systems.The intensity of specific amplified bands of LSV from the different sample parts was more uniform than the intensity of CMV,and the relative contents of CMV was maximum outside of the bulb for the whole viruses infected plants.Our results provide technical support for the detection of virus and virus-free tissue culture and rapid propagation of Lanzhou lily.

关 键 词：兰州百合 黄瓜花叶病毒 百合无症病毒 三重RT-PCR 

分 类 号：S432.1[农业科学—植物病理学] S644.1[农业科学—农业昆虫与害虫防治]