三种百合种球病毒RT-PCR检测

Detection of Three Kinds of Lily Mother Bulb Virus by Using RT-PCR Techniques

本研究以4个百合品种为试材进行种球病毒分子生物学检测。研究结果表明,通过设计兼容性好、性状稳定的三对特异性引物,提高了PCR扩增的效率,改良了RNA的提取方法;改进Trizol法使其适合于内层鳞片及脱毒苗叶片总RNA的提取,而中外层鳞片则采用改进的CTAB法,得到高纯度、高质量以及完整度好的RNA;各反应成分的扩增条件得到了优化,建立了灵敏、快速、低成本的种球病毒RT-PCR检测反应体系及程序。

In this research, four lily cultivars were used as experimental materials to be studied for molecular biological detection of virus in Lily mother bulbs. The results showed that efficiency of PCR by designing three specific primers with good compatibility and stability was improved, as well as the method of RNA extraction was also modified. The improved Trizol method was much more suitable for extracing RNA from lily inner scales and virus-elimination leaves, and the improved CTAB method was better for extracing RNA from middle scales and outer scales with high-purity, good-quality and complete RNA. Therefore, we established a set of virus detection procedures of lily mother bulb by optimizing the PCR amplification conditions, which would be the sensitive, fast and low cost for lily mother bulbs virus detection.