Study On The Diversity, Transmission Mode And Recombination Of Wolbachia Strains In Einong Butterfly Family In China

Wolbachia is a common endosymbiotic bacteria in arthropods and nematodes.It manipulates the reproductive behavior of its host in a variety of ways,including cytoplasmic incompatibility,parthenogenesis,and feminization.(Feminization)and male killing phenomenon.The transmission pattern of Wolbachia is predominantly vertical through the egg cytoplasm and secondarily horizontal from one species to another.Until now,some species of five butterfly groups have been infected with Wolbachia.so far.Despite a few infected butterfly species having been thoroughly studied,there is no systematic study on this infection in Lepidoptera based on an entire tribe.Thus,in this study,we will evaluate the phylogenetic relationship of Aeromachini infected species in China,combining with the diversity of Wolbachia strains to evaluate its transmission mode.In this study,we collected 125 samples from 46 species and 10 genera,and identified all the individual Wolbachia infection.Based on the Multiple sequence typing technology(MLST)and wsp genotyping technology,a comprehensive investigation of the genetic diversity of Wolbachia in the of the Aeromachini in China.The Aeromachini multiple phylogenetic trees were also constructed with both nuclear and mitochondrial genes,as well as multi-locus sequence typing fragments and wsp of Wolbachia.In addition,based on the relaxed molecular clock method,appropriate fossil records are selected as reference points to estimate the origin and differentiation time of the main groups of the Chinese Aeromachini.We compared the age of Wolbachia divergence with divergence times of host species,and estimated the patterns of Wolbachia transmission across this tribe.To examine recombination among Wolbachia strains from Aeromachini species,each MLST gene and wsp gene were detected using RDP3.In the present report we show : 1)of the butterflies examined by diagnostic PCR,25 %(31/125)of samples were Wolbachia positive and 46 %(21/46)of Aeromachini species in this study were considered infected with Wolbachia.2)Nineteen strains of Wolbachia were divided by multi-locus sequence typing(MLST)in this study.According to phylogenetic reconstructions,most of Wolbachia clustered into supergroup B,and the remainings clustered into supergroup A.3)All Wolbachia strains were species specific except for w Jho which was shared by three host species with a sympatric distribution based on a co-phylogenetic comparison between Wolbachia and Aeromachini species.Two tests of phylogenetic congruence did not find any evidence for cospeciation between Wolbachia strains and their butterfly hosts.4)The Bayesian algorithm was used to estimate the divergence time of all Wolbachia-infected species all species of Aeromachini in this study and found that the initial divergences among Aeromachini were about 43 Ma.5)The recombination analysis of each MLST allele and wsp using RDP3 found intragenic recombination in the fts Z gene in four Wolbachia strains.6)The transmission mode of Wolbachia in Aeromachin is not only vertical transmission,but also horizontal transmission.