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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1μg的带毒叶片;实时荧光RT-PCR可分别检测相当于1ng、1ng和100pg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%。
A multiplex RT-PCR and real-time fluorescence RT-PCR for the simultaneous detection of three viruses of Gerbera jamesonii were established by using the gerbera leaves infected with tomato black ring virus, tobacco mosaic virus and cucumber mosaic virus. Multiplex RT-PCR The DNA bands of the three viruses were increased with the detection of 1 μg of infected leaves at the lowest level. Real-time fluorescence RT-PCR was used to detect the infected leaves at 1 ng, 1 ng and 100 pg respectively. At the same time, the virus-free detoxification method was used to detoxify the germ cells inoculated with three kinds of viruses, and the detoxification method was established. The detoxification effects were detected by multiple RT-PCR and real-time fluorescence RT-PCR. The detoxification rates of stem tips were 75.0% and 66.7%, respectively, and the detoxification rates of flowers were 52.0% and 54.5% respectively.