Borah B K, Cheema G S, Gill C K, Dasgupta I
South Campus, Department of Plant Molecular Biology, University of Delhi, Benito Juarez Road, New Delhi, 110021 India.
Indian J Virol. 2010 Oct;21(2):103-9. doi: 10.1007/s13337-010-0018-8. Epub 2011 Jan 11.
A widespread leaf deformity disease of mentha (mint), accompanied by whiteflies, the vectors of begomoviruses, was observed in Punjab in the last few years. The presence of begomovirus was indicated by DNA dot-blot analysis using the conserved coat protein and replication-associated protein genes of another begomovirus, Sri Lankan cassava mosaic virus (SLCMV). A DNA fragment (2.0 kb), representing a partial genomic DNA of a begomovirus, amplified from the symptomatic mentha leaves was used to design end-primers and further amplify an additional 0.9 kb fragment, representing the remaining portion of the resident viral DNA. The two sequences, assembled together (2.7 kb), showed that they represented the complete sequence of an isolate of Tomato leaf curl Karnataka virus (ToLCKV) DNA. Using universal betasatellite primers, a 1.4 kb fragment was amplified from the same sample. This cloned DNA fragment showed complete sequence identity with the previously reported Cotton leaf curl Multan betasatellite (CLCuMB). Majority of the symptomatic mentha leaf samples, collected from four districts of Punjab, showed cross-hybridization in DNA dot-blot using cloned SLCMV and CLCuMB DNA, indicating the presence of one or more begomoviruses related to SLCMV and the betasatellite, CLCuMB. The begomovirus and betasatellite could be mechanically transmitted to Nicotiana benthamiana. Whitefly transmission of the resident begomovirus could also be demonstrated on mentha. The evidence indicates the association of ToLCKV and CLCuMB, a hitherto new combination of a begomovirus and a betasatellite associated with a leaf deformity disease in mentha in Punjab.
在过去几年中,旁遮普邦观察到薄荷出现了一种广泛传播的叶片畸形病害,同时伴有作为双生病毒载体的粉虱。通过使用另一种双生病毒——斯里兰卡木薯花叶病毒(SLCMV)的保守外壳蛋白和复制相关蛋白基因进行DNA斑点杂交分析,表明存在双生病毒。从有症状的薄荷叶片中扩增出一个代表双生病毒部分基因组DNA的2.0 kb DNA片段,用于设计末端引物,并进一步扩增出一个额外的0.9 kb片段,代表驻留病毒DNA的其余部分。将这两个序列组装在一起(2.7 kb),显示它们代表番茄卷叶卡纳塔克病毒(ToLCKV)DNA一个分离株的完整序列。使用通用的卫星DNA引物,从同一样本中扩增出一个1.4 kb的片段。该克隆的DNA片段与先前报道的棉花卷叶木尔坦卫星DNA(CLCuMB)显示出完全的序列同一性。从旁遮普邦四个地区采集的大多数有症状的薄荷叶样本,在使用克隆的SLCMV和CLCuMB DNA进行的DNA斑点杂交中显示出交叉杂交,表明存在一种或多种与SLCMV和卫星DNA CLCuMB相关的双生病毒。双生病毒和卫星DNA可以通过机械接种传播到本氏烟草。驻留双生病毒的粉虱传播也可以在薄荷上得到证实。证据表明ToLCKV与CLCuMB存在关联,这是旁遮普邦一种与薄荷叶片畸形病害相关的双生病毒和卫星DNA的全新组合。