新德里番茄曲叶病毒温州番茄分离物分子鉴定OA

Samples of land tomato with curled and obviously dwarfed leaves were collected from Wenzhou,Zhejiang Province,China.In order to clarify the pathogenic species,RNA of the disease samples was extracted,reverse transcribed into cDNA,and amplified by RT-PCR with the Geminiviridae viral universal primers PA/PB,and its amplified product was identified by sequencing as tomato leaf curl New Delhi virus(ToLCNDV).In order to clarify the evolutionary status of the Wenzhou isolate of tomato leaf curl New Delhi virus,the whole genome sequences of viral DNA-A and DNA-B were obtained by RT-PCR using virus-specific primers.Phylogenetic analysis was performed using Tbtools software,and the results showed that the tomato leaf curl New Delhi virus Wenzhou isolate clustered with the Chinese isolates,which was closest in affinity,and distant from the foreign isolates.The use of virus-encoded proteins to construct transgenic plants is an effective measure to prevent and control viruses.In this study,the genes of AV1(encoding the capsid protein)and BV1(encoding the nucleus shuttling protein)of the Wenzhou isolate of tomato leaf curl New Delhi virus were further cloned,and the transient expression of the tobacco showed that AV1-YFP and BV1-YFP were localized in the nucleus.The results of this study will provide experimental data for further understanding of the distribution of tomato leaf curl New Delhi virus in China,and lay a molecular foundation for the use of virus-encoded proteins to construct transgenic plants for virus prevention and control.