The Impact Of Wolbachia Infection On Gene Expression In Testis And Male Accessory Glands Of Drosophila Melanogaster

Wolbachia pipientis is an obligate intracellular symbiotic strain, found in a wide range of filarial nematodes and arthropods. A great number of arthropods up to70%in insects are infected with Wolbachia. This symbiont manipulates its host’s reproduction via various effects, including male-killing, feminization, parthenogenesis induction (PI) or cytoplasmic incompatibility (CI). CI in all effects is the one more common, which formed a way more conducive to female by altering the offspring survival rate. CI will be induced in the fertilized eggs if infected males and uninfected or different types Wolbachia-infected female mate. Currently there are many researches about Wolbachia, and most of them are on molecular detection and phylogenesis. But the molecular mechanism of CI causing by Wolbachia is unknown.In this study, one hand we picked4genes(CG7194,mdg4,chico and Sgs7) from earlier gene microarray, which got from three instars testis of Wolbachia-infected and uninfected Drosophila melanogaster, and validated by qRT-PCR. The expression of these genes is up-regulated consistent with from microarray. This shows that Wolbachia may affect the early development of fly’s testis. The expression of CG7194is then detected in each development stage of Drosophila through qRT-PCR. In the determination, this gene expresses in whole developmental stages from embryo to3days age of adult fly. The expression level of CG7194in adulthood is higher than others, and it is a partial-male gene. In addition, there is evidence that this gene have a high level in testis according to a paper. So we conjecture that the change of CG7194caused by Wolbachia may have a major impact on male reproductive system of Drosophila. On the other hand, based on the prior outcomes from comparison of proteomics data, we selected6genes whose translation is marked changed in sperm proteins and spermatines. Their expression was tested and compared in testis and male accessory glands of Dmle T and Dmle wMel. The result shows that amount of genes in male accessory glands have no change more than in testis. It could be caused by the regulation of genes during mating briefly or the little impact of genes.4genes (CG7194、Sgs7、CG11037and lectin-30A) are selected from above two detections, and successful got their vectors. Through microinjection technique these vectors were injected into earlier developmental fertilized eggs. Then the function of these genes could be studied by selected transgenic flies.