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期 刊 ：植物保护学报 2009年

关键词：番茄黑环病毒(TBRV)；烟草花叶病毒(TMV)；黄瓜花叶病毒(CMV)；RT-PCR；非洲菊脱毒；

Keywords:Tomato black ring virus, Tobacco mosaic virus, Cucumber mosaic virus, RT-PCR, virus-free for Gerbera jamesonii,

摘 要 ：利用感染了番茄黑环病毒、烟草花叶病毒和黄瓜花叶病毒的非洲菊叶片,建立同步检测非洲菊3种病毒的多重RT-PCR和实时荧光RT-PCR,多重RT-PCR可一次性扩增出3种病毒的DNA条带,最低可检测到相当于1μg的带毒叶片;实时荧光RT-PCR可分别检测相当于1ng、1ng和100pg的带毒叶片。同时采用茎尖、叶片和花托组织培养脱毒并结合培养基添加病毒A的脱毒方法,对接种3种病毒的非洲菊进行外植体脱毒试验,获得非洲菊组培苗脱毒方法。用建立的多重RT-PCR和 实 时 荧光RT-PCR检测脱毒效果, 茎尖脱毒率分别为75.0% 和66.7%, 花托脱毒率分别为52.0%和54.5%。

Abstract:We developed multiplex RT-PCR and real-time RT-PCR with Gerbera jamesonii Bolus infected Tomato black ring virus, Tobacco mosaic virus and Cucumber mosaic virus. We could detect the virus as low as 1μg leaf tissue by multiplex RT-PCR and as low as 1ng, 1ng and 100pg the three leaf tissues respectively by real-time RT-PCR. We used several virus-free by tissue culture for the study on Gerbera jamesonii infected these 3 viruses for the first time, verified the effect of virus-free by multiplex RT-PCR and real-time RT-PCR, found that the virus-free rates of tip culture were 75.0% and 66.7% respectively, the virus-free rates of torus culture were 52.0% and 54.5% respectively.

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