ATP6蛋白影响异沙叶蝉对小麦蓝矮植原体的传播效率

ATP6 protein affects the transmission efficiency of wheat blue dwarf phytoplasma by the leafhopper Psammotettix alienus(Hemiptera:Cicadellidae)

【目的】探索异沙叶蝉Psammotettix alienus ATP6蛋白对小麦蓝矮植原体(wheat blue dwarf phytoplasma,WBDp)传播效率的影响。【方法】将异沙叶蝉mRNA反转录合成cDNA,构建分离泛素酵母双杂交膜系统cDNA文库;用WBDp的虫传相关蛋白P2-4为诱饵,从异沙叶蝉cDNA文库杂交筛选互作蛋白;通过RNAi沉默ATP6基因,验证ATP6蛋白对异沙叶蝉3龄若虫传播WBDp的影响。【结果】构建了异沙叶蝉分离泛素酵母双杂交膜系统cDNA文库,用P2-4从cDNA文库中杂交筛选获得了ATP6等多种互作蛋白。RNAi结果发现沉默ATP6基因后异沙叶蝉3龄若虫的WBDp传播率比对照组显著下降了23.33%±3.33%,表明ATP6是异沙叶蝉参与WBDp传播的关键蛋白。【结论】明确了异沙叶蝉ATP6是参与WBDp传播的一个关键蛋白。为进一步探明异沙叶蝉传播WBDp的分子机制和植原体病害的田间防治提供了理论基础。

【Aim】To investigate the effect of ATP6 protein of the leafhopper Psammotettix alienus on the transmission efficiency of wheat blue dwarf phytoplasma(WBDp).【Methods】cDNA was synthesized by reverse transcription from the mRNA of P.alienus,and then a cDNA library was constructed with cDNA using a split-ubiquitin yeast two-hybrid membrane system.Protein P2-4 related to the transmission of WBDp was used as bait to screen interacting proteins by hybridizing with the cDNA library of P.alienus.The effect of ATP6 protein on the WBDp transmission by the 3rd instar nymphs of P.alienus was verified by RNAi of ATP6 gene.【Results】A cDNA library of P.alienus using a split-ubiquitin yeast two-hybrid membrane system was constructed.Several interacting proteins including ATP6 were obtained in cDNA library using P2-4 as the bait.Silencing of ATP6 gene by RNAi significantly reduced the transmission rate of WBDp by the 3rd instar nymphs of P.alienus by 23.33%±3.33%as compared with the control group,indicating that ATP6 is a key protein involved in WBDp transmission by P.alienus.【Conclusion】ATP6 of P.alienus has been identified as a key protein involved in the transmission of WBDp.This study provides a theoretical basis for further understanding the molecular mechanisms of WBDp transmission by P.alienus and controlling phytoplasma diseases in fields.