Diseases caused by begomoviruses are becoming the major limiting factors for the production of watermelon in India. Survey for the incidence of plants showing symptoms typical to begomovirus infection was conducted in watermelon fields. The study revealed that 40% of the watermelon plants were showing the yellowing and downward curling symptoms. Twenty infected samples were collected from the different farmer's fields to know the association of begomoviruses. The PCR amplification using begomovirus-specific primers resulted in an expected 1.2 kb PCR product indicating the begomovirus association with the watermelon samples. The sequence comparison results of 1.2 kb representing partial genome revealed that all sequences obtained from watermelon samples have a nucleotide (nt) identity of more than 98% among them and are maximum homology with Tomato leaf curl New Delhi virus (ToLCNDV). One watermelon sample (WM1) was selected for complete genome amplification using RCA method (rolling-circle amplification). Amplification of DNA B and no amplification of betasatellites and alphasatellite indicated this virus as bipartite. Sequence Demarcation Tool (SDT) analysis of the DNA A component of the WM1 isolate showed the maximum nt identity of 94.6-97.9% and 85.2-95.8% with ToLCNDV infecting cucurbits. The recombinant analysis showed that the genome was likely to be derived from the recombination of already reported begomoviruses (ToLCNDV, ToLCPalV, and MYMIV) infecting diverse crops. The whitefly cryptic species predominant in the begomovirus-infected watermelon fields were identified as Asia-II-5 group. The LAMP assay developed based on coat protein gene sequence was able to detect the ToLCNDV in the infected samples. Visual detection of the LAMP-amplified products was observed with the hydroxy naphthol blue. LAMP assay was also validated with ToLCNDV infected sponge gourd, spine gourd, ivy gourd, ridge gourd, and cucumber. This is the first report of ToLCNDV association with leaf curl and yellowing disease of watermelon from India and World based on complete genome sequencing.

译文

由beg病毒引起的疾病正在成为印度西瓜生产的主要限制因素。在西瓜田中进行了显示出典型贝古病毒感染症状的植物发病率的调查。研究表明,40% 西瓜植物都表现出发黄和向下卷曲的症状。从不同农民的田地收集了20个受感染的样本,以了解begomovirus的关联。使用begomovirus特异性引物的PCR扩增产生预期的1.2 kb PCR产物,表明begomovirus与西瓜样品的关联。代表部分基因组的1.2 kb的序列比较结果表明,从西瓜样品中获得的所有序列具有超过98% 的核苷酸 (nt) 同一性,并且与番茄卷曲新德里病毒 (ToLCNDV) 具有最大的同源性。使用RCA方法 (滚动环扩增) 选择一个西瓜样品 (WM1) 进行完整基因组扩增。DNA B的扩增和 β 卫星和 α 卫星的未扩增表明该病毒为二分病毒。WM1分离物的DNA A组分的序列分界工具 (SDT) 分析显示,与感染葫芦的ToLCNDV具有94.6-97.9% 和85.2-95.8% 的最大nt同一性。重组分析表明,基因组很可能来自已经报道的begomovirus (ToLCNDV,ToLCPalV和MYMIV) 感染多种作物的重组。在begomovirus感染的西瓜田中占主导地位的粉虱隐种被确定为Asia-II-5组。基于外壳蛋白基因序列开发的LAMP测定法能够检测感染样品中的ToLCNDV。用羟基萘酚蓝观察到灯放大产物的视觉检测。LAMP试验也用ToLCNDV感染的海绵葫芦、脊葫芦、常春藤葫芦、脊葫芦和黄瓜进行了验证。这是基于完整基因组测序的ToLCNDV与印度和世界西瓜的叶卷曲和黄化病的首次报道。

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