| 【Objective】The response of SmWRKY65（Solanum melongena L. WRKY65）to bacterial wilt of eggplant was explored by cloning SmWRKY65 gene and analyzing its bio-information and function.【Method】The eggplant disease-resistant plants（E-31）and cDNA from its leaves were used as tested materials and template, respectively. The SmWRKY65 was obtained by PCR technology and bioinformatics analysis was constructed by ExPASy ProtParam tool. The expression analysis of different tissue sites and disease-resistant and susceptible materials（E-32）was performed by quantitative real-time PCR（qRT-PCR）method. Subcellular localization vector was constructed by double digestion method. The pTRV2-SmWRKY65 vector was constructed by virus induced gene silencing（VIGS）technology and the disease-resistant eggplant plants were infected.【Result】The SmWRKY65 gene was successfully cloned in eggplant. The gene contains 834 bp nucleotide sequences, codes 277 amino acid residues in its open reading frame, and concludes a WRKY conserved domain containing 60 amino acid in the regions of 71-131. The analysis of subcellular localization showed that SmWRKY65-GFP fusion protein was found in the cell nucleus. The expression analysis of different tissues revealed that SmWRKY65 had highest expression in eggplant roots, followed by leaves and stems. The qRT-PCR analysis testified that SmWRKY65 was up-regulated in response to the induction of bacterial wilt in eggplant resistant and susceptible plants. The VIGS results showed that the leaves of pTRV2-SmWRKY65 silent plants was observed clear wilting symptoms when compared
with water and pTRV2 control vector. By qRT-PCR profiling, it was observed that pTRV2-SmWRKY65 expression was obviously decreased when compared with control groups.【Conclusion】The expression level in disease-resistant eggplant materials was significantly higher than that in susceptible materials, and the pTRV2-SmWRKY65 plants showed more symptoms of susceptibility when compared with control groups, indicating that the silence of SmWRKY65 reduced the resistance to bacterial wilt of eggplants and SmWRKY65 was possibly related to the process regulation of eggplant resistance to bacterial wilt. |
