显微注射青霉素G获得单感染Wolbachia和单感染Cardinium白背飞虱品系

Wolbachia和Cardinium都是广泛存在于节肢动物体内的一类母系遗传的共生细菌， 可以通过不同方式操纵寄主的生殖行为。Wolbachia和Cardinium感染同一寄主在自然界比较常见， 但是在某些可以同时感染Wolbachia和Cardinium的寄主中其单感染品系较难发现。本研究检测了云南文山(YN)、 海南三亚(HN)这2个不同地理种群中Wolbachia和Cardinium的感染情况； 以双感染Wolbachia和Cardinium的白背飞虱Sogatella furcifera海南种群为实验材料， 运用显微注射方法对双感染Wolbachia和Cardinium的白背飞虱若虫注射不同浓度青霉素G以获得单感染品系。结果表明： 白背飞虱自然种群中单感染Wolbachia比率极低， 本实验用到的海南种群未检测到单感染Wolbachia成虫； 通过显微注射青霉素G的方法可以从白背飞虱双感染品系中筛选获得单感染品系， 当青霉素G注射浓度为0.2%(w/v)， 注射龄期为5龄时得到单感染品系效率最高； F5代的检测结果显示显微注射得到的单感染品系可以稳定遗传。本研究为获得单感染品系白背飞虱提供了快捷方法， 同时为其他双感染Wolbachia和Cardinium节肢动物不同感染品系的筛选提供参考。

Establishment of singly Wolbachia- and singly Cardinium-infected whitebacked planthopper ( Sogatella furcifera ) lines by microinjecting penicillin G

Wolbachia and Cardinium are both maternally inherited intracellular bacteria that infect a wide range of arthropods and are associated with various reproductive abnormalities in their hosts. Infection with both Wolbachia and Cardinium is reasonably common, yet it is relatively difficult to find the individuals infected with single symbiont in doubly infected hosts naturally. In this study, Wolbachia and Cardinium were detected in two populations of the whitebacked planthopper, Sogatella furcifera (Horváth), collected from Wenshan of Yunnan (YN) and Sanya of Hainan (HN). Using the Hainan population of doubly infected S. furcifera as the material, we developed a new technique of microinjecting different concentrations of penicillin G into nymphs of the planthopper. The results showed that the rate of individuals infected with only Wolbachia was very low in natural populations of S. furcifera, and no singly Wolbachia-infected individual was found in the Hainan population. We got singly-infected individuals by using microinjection of penicillin G in the laboratory. The singly infected lines can be produced most efficiently by injecting 0.2% (w/v) penicillin G into the 5th instar nymphs. The result of PCR detection of F5 progenies showed that the singly infected lines obtained by microinjection can be inherited stably. These results here will provide a speedy method to get singly infected lines, and this method can be used to establish lines with different infection type for other arthropods doubly infected with Wolbachia and Cardinium.