The Relation Between The Wolbachia In Drosophila Melanogaster Of Three Areas In China And The Influence Mechanism Analysis Of Wolbachia On Host Reproduction

The wolbachia is a kind of common intracellular symbiotic bacterial of arthropods and nematodes. It is a kind of gram negative bacteria,matrilineal interitance, the amounts of the arthropod species infected with wolbachia is about70%in the world wide. The geographical conditions are complicated in our country, and the area is wide, but the studies that the infection of wolbachiain Drosophila melanogasterof our country are very few. Wolbachia can manipulate the reproduction of host to improve its transmission, the ways of manipalative including cytoplasmic incompatibility(CI), killing male offspring,feminising and parthenogenesis. And CI is the best konwn phenotype of these alterations. CI occurs when the wolbachia-infected male host mate with the wolbachia-uninfected female host, or the wolbachia-infected male host mate with the female host which is infected with different kinds of wolbachia. It will result in low rate of viable offspring or no hatch. But the female mate with the m.ale infect the same specie wolbachia or uninfected male can protece the embryos of normal development. At present, the researches about CI are very mach,but the exact mechanism of CI remains unclearly. We detected the relationship of the wolbachia in the Drosophila melanogasterof our country3areas, then studied the influence of Wolbachia to Drosophila melanogaster reproductive and the mechanism.1、our research group has previously detected Wolbachiainfection of six distinct geographical populations of Drosophila melanogaster in China, we found that populations from Wuhan in Hubei Province, Liuku in Yunnan Province and Tianjin are infected by Wolbachia. We analysis the gene of wolbachia which are infecting the Drosophila melanogaster of the3place through MLST. It shows that the Drosophila melanogaster of the3place belong to different kinds of ST. The ST of wolbachia which is infecting Drosophila melanogaster of wuhan is398,and the wolbachia of tianjing is397, liuku is1. The ST of wuhan and tianjing are new emergence.In order to further realize the relationship between the wolbachia of the3places,and the influence to flies reproduce, We detected the CI of the wolbachia which are infecting theDrosophila melanogaster of hubei wuhan, yunnan liuku, tianjing. the results showed that the CI of the wolbachia infected wuhan and tianjing files are very strong, but the CI of liuku is weak. Beacause the distance of3areas is far, so it is the reason of the different CI the different files. We explore the relationship between the fruit files in3regions through crossing. The result shows that the genetic relationship is more close between the fruit flies of wuhan and tianjing. And the genetic relationship is further between the fruit flies of wuhan and liuku. We research the relationship between the3region wolbachia through crossing, and statisticing the hatching rate of offspring. The result shows that the wolbachia in WuhanDrosophila melanogaster could not rescue the CI phenotype which was caused by wolbachia in Tianjing Drosophila melanogaster. And the wolbachia in Tianjing Drosophila melanogaster could not rescue the CI phenotype which was caused by wolbachia in Wuhann Drosophila melanogaster. The results showed that there are some influences to the evolution of Drosophila melanogaster because of wolbachiamutualistic symbiosis.2、our research group has previously detected the CG10433expression quantity in testis of third instar larvae of Dmel wMel (wolbachia-infected) is higher than Dmel T (wolbachia-uninfected) by microarry. And we By quantitative RT-PCR(qRT-PCR), and we has obtained the fruit flies that has induced overexpression of CGI0433. We found the expression levels of gene CGI0433were significantly increased in CGI0433-overexpression Drosophila by quantitative RT-PCR(qRT-PCR). We tested the CI strength in1-day-old and5-day-old CGI0433-overexpression Drosophila to detect whether the CI phenotype exists. The result of crossing experiments shows that1-day-old CG10433-overexpressionDrosophila can induce CI phenotype. Another the mating between the CGI0453-overexpression male with Dmel T female resulted in that the laying amount of the female decrease. It may be the reason why the number of offspring decrease in the CI mating group. But CG10433-overexpression could not effect the fertility of female,because the embryo hatching rate after CG10433-overexpression female mating with Dmel T male is normal. The results showed that the gene CG10433expression quantity in the flies testis increase may be the one of reasons of CI.