The Study On Subcellular Localization Of Citrus Tatter Leaf Virus Coat Protein And Movement Protein In Nicotiana Benthamiana

Citrus tatter leaf virus (CTLV) is one of the most important pathogens that infect citrus induced leaf malformation. It is mainly distributed in citrus production areas in China and some other counties throughout the world, resulting in remarkable economic losses in both yield and quality. Today researchs about CTLV focus on the molecular biological characteristic and genome expression strategy but the virus infection cycle and not clear.Coat protein gene and movement protein gene are two main functional genes of CTLV and play an important role in the process of virus manipulation. In order to identify the subcellular localization of both CP and MP genes, The present study laid the foundation and provided clues for further functional analysis of CP and MP genes.The obtained results are as follows:1. PCR was used to amplify these two genes from CTLV-STO (GenBank accession numbers JQ765412) infectious clone plasmid,Then cloned. Gateway technology was applied to construct plant expression vectors of both CP and MP genes subsequently transformed to agrobacterium tumefaciend strain GV3101by heat shock method.2. Transient expression was performed via Agroinfection assay in Nicotiana.benthamiana. Post three days after infiltration, N.benthamiana leaves expressed the fused protein containing target genes and fluorescent drived from the vector were visualized by laser cofocol scanning microscopy. Agroinfection assay followed by laser confocal microscopy analysis showed that the subcellular localization of CTLV CP localized both in the nucleus and cell membrane (or cell wall).For the CTLV MP the subcellular localization mainly in cell membrane (or cell wall). To confirm this result, two vectors with different fluorescent tag were tested.The localization of CTLV CP in N.benthamiana leaves were consistent even with different vectors.