贵州芸薹黄化病毒的分子检测及基因型鉴定

Molecular detection and genotype identification of Brassica yellows virus in Guizhou

  • 摘要: 【目的】对贵州省白菜、萝卜和甘蓝感染芸薹黄化病毒(Brassica yellows virus,BrYV)进行分子检测及基因型鉴定,为马铃薯卷叶病毒属病毒的防治提供理论参考。【方法】在贵州省遵义市、威宁县、关岭县、平坝县等县(市)共采集284份蔬菜(白菜、萝卜和甘蓝)疑似病毒病样品,利用马铃薯卷叶病毒属的通用引物进行RT-PCR分子检测,并对检测出的16个BrYV分离物进行P0CP基因序列扩增,再与已报道的BrYV基因序列进行比对,从而构建系统发育进化树,鉴定其基因型。【结果】284份疑似病毒病样品中,有43份样品检测出BrYV,检出率为15.14%,其中,感染BrYV白菜样品12份,占白菜总样品数的12.77%;感染BrYV萝卜样品14份,占萝卜总样品数的20.29%;感染BrYV甘蓝样品17份,占甘蓝总样品数的14.05%。贵州16个BrYV分离物P0基因核苷酸序列的相似性为89.9%~100.0%,CP基因核苷酸序列的相似性为93.5%~100.0%,二者编码的氨基酸序列相似性均为100.0%。基于P0基因构建的系统发育进化树可知,BrYV-Pingba-BC-2、BrYV-Zunyi-LB-1和BrYV-Zhijing-BC-1均与BrYV-BJS和BrYV-BBJ聚为一类,其余13个分离物均与BrYV-ABJ和BrYV-AJS聚为一类。基于CP基因构建的系统发育进化树可知,贵州不同地区不同蔬菜作物的16个BrYV分离与BrYV-A和BrYV-B基因型无明显分界,亲缘关系均较近。BrYV P0基因检测到5个重组事件,BrYV CP基因未检测到潜在重组事件。【结论】BrYV在贵州白菜、萝卜和甘蓝上普遍发生,但对萝卜的危害最重,主要存在BrYV-A和BrYV-B 2种基因型。

     

    Abstract: 【Objective】Molecular detection and genotype identification of Brassica yellows virus(BrYV) infection in Chinese cabbage, radish and cabbage in Guizhou were conducted to provide reference for control of potato leaf curl virus.【Method】284 samples of vegetable(Chinese cabbage, radish and cabbage) virus diseases were collected in Zunyi, Weining, Guanling, Pingba and other cities(counties) in Guizhou. RT-PCR molecular detection using the common primers of potato leaf curl virus was conducted and P0 and CP gene sequences amplification was conducted with the 16 BrYV isolates detected, then compared to the reported BrYV gene sequences. Phylogenetic tree was constructed and its genotype was identified.【Result】BrYV was detected in 43 suspected samples out of 284 virus disease samples, and the positive rate was 15.14%. Among them, there were 12 BrYV infected samples in cabbage samples, accounting for 12.77% of the total number of cabbage samples;14 BrYV infected samples in radish samples, accounting for 20.29% of the total number of radish samples;17 BrYV infected samples in cabbage samples, accounting for 14.05% of the total number of cabbage samples. The nucleotide similarity of P0 gene and CP gene of 16 BrYV isolates were 89.9%-100.0% and 93.5%-100.0%, respectively. The consistency of amino acids sequence was 100.0%. Phylogenetic tree based on P0 gene showed that, BrYV-Pingba-BC-2, BrYV-Zunyi-LB-1, BrYV-Zhijing-BC-1, BrYV-BJS and BrYV-BBJ were clustered into one branch, and the other 13 isolates were clustered into one branch with BrYV-ABJ and BrYV-AJS. There was no obvious boundary between 16 BrYV isolates and BrYV-A and BrYV-B genotypes from different vegetable crops in different areas of Guizhou according to Phylogenetic tree based on CP gene, and the genetic relationships were all relatively close. Five recombination events were detected in BrYV P0 gene and no potential recombination events were detected in BrYV CP gene.【Conclusion】BrYV is prevalent in Chinese cabbage, radish and cabbage in Guizhou, but the damage radish is the most serious, there are two genotypes of BrYV-A and BrYV-B.

     

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