Identification And Functional Analysis Of Interacting Proteins With CP Of Ramie Mosaic Virus In Bemisia Tabaci

As the only known vector of Begomoviruses,whitefly Bemisia tabaci play a critical role in the transmission of these viruses.Begomoviruses are transmitted by the B.tabaci in a persistent circulation manner.Many evidence have suggested that the coat protein(CP)of Begomovirus is important in the vector transmission.The different interaction between virus and whitefly exerts positive,negative or neutral determining by the different combinations of whiteflies,begomoviruses and plants.However,little has been known about the molecular mechanisms of begomoviruses transmission by B.tabaci.Here,the new begomovirus Ramie mosaic virus(RaMoV),the genus Begomovirus in the family Geminiviridae,is dominant in China and transmission by B.tabaci.In this research,Ramie mosaic virus(RaMoV)and its vector B.tabaci MED were used as a model to study direct and indirect interaction and virus transmission by B.tabaci MED of different sexes.The transcriptional response of the B.tabaci MED species to RaMoV was studied with illumina sequencing.The whitefly receptor proteins interacting with RaMoV coat protein were screened and identified.The main results were as following:1.No significant direct or indirect effects of RaMoV were observed in B.tabaci MED females,although RaMoV could shorten the life span of B.tabaci MED females by up to 4 days.The tests of RaMoV acquisition and transmission by different sexes of B.tabaci MED showed that there were higher virus acquisition and transmission efficiency by females than males.It means that RaMoV is transmitted by B.tabaci MED in a sex-dependent manner.2.The viruliferous and non-viruliferous of B.tabaci MED species were selected as the research targets with transcriptome sequencing.The underlying interaction mechanism between RaMoV and B.tabaci was explored and analyzed.A large number of candidate genes involved in virus and vector interaction relationship were acquired.Such as the possible genes associated with glucose metabolism,detoxification metabolism and autophagy pathways.These indicates that the insect target proteins interacted with RaMoV CP and participates in the process of virus acquisition and transmission.3.Yeast two hybrid system had be used to screen proteins in the whitefly interacting with RaMoV CP.Then,yeast two hybrid and GST pull-down will be used to confirm the proteins interaction in vivo and in vitro.The ZNF330 protein of B.tabaci was verified interaction with RaMoV CP protein.ZNF330 proteins were highly conservation in phylogeny and were present throughout evolution from nematode to humans.4.The functions of ZNF330 protein were analyzed by bioinformatics analysis with the signal P-NN,transmembrane helices,glycosylation sites,hydrophobicity or hydrophilicity in B.tabaci.Relative expression of ZFP330 m RNA under RaMoV treatment of B.tabaci by q PCR show positive correlation with the increase of RaMoV DNA accumulation in whiteflies.The functional analyses of ZNF330 in the interaction between insect vector and virus will be identified by RNAi technology.These results suggest that the amounts of virus in ZNF330 silenced whiteflies are significant increase.In conclusion,the results in this research had revealed the transmission ability of B.tabaci to begomoviruses and the interaction between ZNF330 protein in B.tabaci and RaMoV CP protein.The biological function of ZNF330 protein were also studied in this research.This results would provide new theoretical basis and thoughts for further understanding transmission mechanisms of B.tabaci and control of ramie mosaic disease.