Identidication And Characterization Studies Of Wolbachia Effectors With ANK Domains In Trichogramma Dendrolimi

Wolbachia is the most widely distributed and abundant group of symbiotic bacteria in arthropods.Wolbachia can modify changesof host reproductive phenotypes,including cytoplasmic incompatibility(CI),feminization(F),male killing(MK),and parthenogenesis(PI).It is considered to be closely related to evolutionary biological issues such as sex determination,and is expected to have broad prospects in biological control programs against pests.At present,the CI mechanisms of the interaction between Wolbachia and hosts have been studied well.Howerver,and the mechanisms on Wolbahcia induced PI phenotype are poorly understood.ANK domains are a group of certain sequences encoding protein-to-protein interactions,capable of performing a various biological functions,playing an important role in host-microbial interactions and infection evolution.To reveal the molecular mechanisms of Wolbachia regulating the PI phenotype of hosts,this study identified the Wolbachia effector proteins containing the ANK domain in Wolbachia-infected Trichogramma dendrolimi strain,The subcellular localization and expression patterns of these proteins were conducted.The results are as follows:1.Gene cloning of Wolbachia effector proteins containing the ANK domain.This study identified 40 ANK-containing proteins gene in the Wolbachia genome that cause PI in Trichogramma dendrolimi.38 protein gene sequences were identified by gene cloning.2.Wolbachia effector proteins containing ANK was screened for induced expression.Total 33 recombinant plasmids were constructed.The functions of candidate proteins were determined by the growth inhibition in yeast with two pressure sources(sodium chloride and sorbitol).Four effector proteins were identified by the significant inhibition on yeast growth.3.Subcellular localization containing the ANK domain Wolbachia effector proteins.To determine the action of the effectors inner the cells,the subcellular localization of effectors was observed in the yeast cell.The results showed that the effector proteins were all localized on the nucleus of yeast cells.4.Analysis of Wolbachia effector proteins expression patterns containing ANK.RT-q PCR was applied to detect the expression levels of four effector proteins containing ANK domains at different stages of host Trichogramma(larval,pre-pupae,pupae,adult).The results showed that the expressions of four effector proteins were not different among different stages of host Trichogramma.The identification of effector proteins is the first step to gain a further understanding of Wolbachia-induced PI in host Trichogramma dendrolimi.This study lays a certain foundation for the exploration of the interaction mechanism between Wolbachia and the host.