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为明确黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)在葫芦上的种传规律,以西瓜砧木葫芦种子为材料,采用双抗体夹心酶联免疫吸附测定法(DAS-ELISA)、反转录聚合酶链式反应(RT-PCR)与生物学检测相结合的方法研究葫芦种子的带毒率和传毒率的关系,并评价了干热处理对病毒的钝化效果。结果表明,DAS-ELISA灵敏度检测种子时,在感染种子研磨液∶健康种子研磨液为1∶1 000时,带毒量仍能检测出阳性,RT-PCR和DAS-ELISA两种方法均能准确检测葫芦种子的带毒情况;6个批次的葫芦种子有4个批次呈阳性,带毒率在0~100%之间,贮藏1年后的传毒率在0~5.6%之间;4个为CGMMV阳性的种子批经干热处理后,仅1株实生苗呈阳性。研究表明,CGMMV在葫芦作物上的隐性带毒现象非常普遍;种子的带毒率高而传毒率低,以表面带毒为主,且非常稳定;72℃72 h干热处理葫芦种子能有效地钝化CGMMV。
In order to clarify the law of seed germination of cucumber green mottle mosaic virus (CGMMV) on gourd, the water hyacinth rootstock gourd seed was used as material and the double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) Transcriptional Polymerase Chain Reaction (RT-PCR) and biological detection methods were used to study the relationship between the virulence rate and the virus transmission rate of gourd seeds, and the passivation effect of dry heat treatment on the virus was evaluated. The results showed that when the sensitivity of DAS-ELISA was tested, the virulence of the infected seed slurry was still positive when the seed slurry was inoculated at 1: 1,000, and both RT-PCR and DAS-ELISA were accurate The gourd seeds were tested for the poisoning status. Four batches of gourd seeds from six batches were positive with the rate of 0-100%, and the rate of one year after storage was 0-5.6%. Four of the CGMMV-positive seeds were approved for dry heat treatment and only one was positive for seedlings. The results showed that the CGMMV was very common in the gourd crops. The high virulence rate of the CGMMV and the low rate of virus poisoning on the surface of the gourd were mainly caused by the surface poisoning, and the seeds were stable at 72 ℃ for 72 h. Passivating CGMMV.