关于二羧酸-三羧酸盐载体蛋白负调控黄瓜花叶病毒积累的初步研究

Preliminary study on the negative regulation of Cucumber mosaic virus accumulation by dicarboxylate-tricarboxylate carrier

黄瓜花叶病毒(Cucumber mosaic virus,CMV)的复制场所位于液泡膜,但是参与调控CMV复制的寄主蛋白因子有待进一步研究。植物二羧酸-三羧酸盐载体蛋白(Dicarboxylate-tricarboxylate carrier,DTC)为线粒体膜和液泡膜定位的跨膜蛋白,参与二羧酸盐和三羧酸盐的跨膜转运。为了探索DTC在CMV复制中的调控作用,通过基于烟草脆裂病毒(Tobacco rattle virus,TRV)的基因载体下调本氏烟体内DTC2(NbDTC2)的mRNA水平,并通过GFP荧光和Western blot分析DTC2下调表达后对CMVΔ2b-eGFP积累量的影响,发现NbDTC2的下调表达增加CMV的积累水平;同时分析本氏烟DTC2的瞬时过表达对CMVΔCP-eGFP积累的影响,通过观察eGFP荧光和Western blot检测,发现NbDTC2的过表达在一定程度上抑制病毒的积累。以上结果表明:DTC2负调控CMV的积累,可能参与植物对病毒的防御。

The replication site of Cucumber mosaic virus （CMV） is located on vacuolar membrane, but the host protein factors involved in CMV replication need further study. PlantDicarboxylate tricarboxylate carrier （DTC） is the transmembrane protein that locates the mitochondrial membrane and vacuole membrane, and is responsible for transportation of dicarboxylate and tricarboxylate. To investigate the regulation role of DTC in CMV replication, mRNA level of DTC2 （NbDTC2） in Nicotiana benthamian was lowered through the genophore of Tobacco rattle virus （TRV）. The influence of CMVA2b eGFP accumulation after down regulation of DTC2 was analyzed with GFP fluorescence and Western blot. It was found that, the down regulation of NbDTC2 increased the accumulation level of CMV. Meanwhile, the influence of transient overexpression of DTC2 on accumulation of CMVACP eGFP was analyzed. The results of GFP fluorescence and western blot showed that overexpression of NbDTC2 inhibited accumulation of the virus to some extent. The above results indicate that, DTC2 negatively regulates CMV accumulation, which may participate in the defense against CMV.