Divergence Of Wiebesia Spp. And Their Infection Of Intracellular Bacteria Wolbachia

In the researches before, Webesia spp. distributed in China mainland have divergenced into3cryptic species. Geographic events may probably be the reason of their speciation. With people know more and more about intracellular bacteria Wolbachia, it was thought that Wolbachia may lead to speciation of their hosts in some instance. In some studies, different fig wasps cryptic species were found harbored different Wolbachia strains.Analysis of female samples which were distributed in Japan showed deep divergence of mtDNA gene COI and lower divergence in nuclear gene EFl-il. Neighbor-joining, maximum likelihood and Bayesian tree all cluster Webesia spp. samples into4clades with high bootstrap and posterior probability. Japanese Webesia spp. were phylogenetically closer to Webesia sp.3. Interclade phylogenetic distances are10times to the intraclade ones, thus pollator of F. pumila can be identified as4cryptic species.In this study432Webesia spp.(the pollinator of Ficus pumila) from41populations were test and found that only Webesia sp.l harbor Wolbachia with100%infection rate. What's more, sequencing of gene wsp, AsZand16S rRNA showed that only one Wolbachia strain was held in all25tested populations of Webesia sp.l. The results of BLAST showed that the Wolbachia strain which have the same wsp gene infect insects which are not fig wasps. This may suggest that Webesia sp.l harbor Wolbachia through horizontal transmission.Webesia sp.l cluster sp.2and sp.3cluster Japanese samples, divergence time are about3million years ago. These time are coincident with the launch of Wuyi Mountain and the form of the Ryukyu Islands. Webesia spp. are temperature and altitude sensitive species with limited flying ability. Thus considering only Webesi'a sp.l harbor Wolbachia, geographic events may weighted greatly in the speciation of Webesia spp. Actually Webesia sp.l may infected by Wolbachia after speciation.According to DNA barcoding of mtDNA COI gene and nuclear EFla gene identified the female non-pollinating wasps in the syconia of F. pumila as Philotrypesis, which agrees with the morphological identification. Through Wolbachia infection test, it was found that Philotrypesis sp. which were only found dwelling with Wiebesia sp.2harbor the same Wolbachia strain with the Webesia sp.l. All5Philotrypesis sp. samples got positive results of infection test which suggests the most recent common ancestor of sp.l and sp.2probabliy harbor Molbachia. Infection test also showed that Wiebesia sp.2may immune from Wolbachia.