生姜脱毒快繁与病毒检测技术研究

Study on Virus-free and Detection Technology of Ginger

本文以感染TMV和CMV的湘西本地小黄姜为试材,采用热处理、茎尖培养结合添加病毒唑的方法进行脱毒处理以获得无病毒的优质生姜种苗,运用RT-PCR方法检测病毒,以期探讨出生姜最适宜的脱毒快繁及病毒检测技术。结果表明,将生姜块茎置于40℃条件下催芽,显微镜下剥取0.2~0.5 mm的茎尖进行培养,最佳诱导培养基的配方为MS+6-BA 2 mg/L+NAA 0.5 mg/L+病毒唑40 mg/L,继代培养基中将MS培养基中的蔗糖浓度调整为80 g/L,即可实现生姜试管苗的增殖、生根及壮苗的一步培养,培养周期以30~40 d为宜。建立了一套生姜CMV和TMV 2种病毒的RT-PCR检测体系,运用该检测方法对生姜脱毒试管苗进行检测,病毒检出率为0,生姜脱毒率达100%。

In order to obtain high quality ginger seedlings without virus,the yellow ginger of Xiangxi three main virus detoxification methods werestudied.Taking TMV and CMV infection Yellow ginger of Xiangxi as experimental materials,the heat treatment,stem tip cultivation method of addingribavirin combination,in the process of test using RT-PCR method to detect virus,the most suitable detoxification and virus detection technology ofginger were discussed.The results showed that while putting the ginger tuber for germination at 40 ℃,and then wring 0.2-0.5 mm size stem tip underthe microscope for culturing.The optimal medium for induction was MS＋6-BA 2 mg/L＋NAA 0.5 mg/L＋virazole 40 mg/L.In the subculture medium,when the sucrose concentration in the MS medium was adjusted to 80 g/L,the ginger test tube忆s proliferation,taking root and strong seedlings can berealized by one step,and the suitable culture cycle is 30-40 days.A RT-PCR detection system for CMV and TMV viruses was established.The virusdetection rate of ginger detoxification test tube was zero by RT-PCR,ginger detoxication rate is 100%.