灰飞虱体内沃尔巴克氏体的检测

Molecular Detection of Wolbachia in Small Brown Planthopper(Laodelphax striatellus)

采用沃尔巴克氏体(Wolbachia)表面蛋白基因(wsp基因)的通用引物81F和691R对河北曲阳和容城灰飞虱(Laodelphax striatellus)体内的Wolbachia进行PCR检测,结果表明,从灰飞虱DNA样品中可扩增出600 bp大小的wsp目的基因片段,证实河北曲阳和容城田间的灰飞虱种群有Wolbachia感染;对wsp目的基因片段的序列测定和同源性分析表明,河北的2个灰飞虱种群感染的Wolbachia与来自白背飞虱的wFur品系、灰飞虱的wStri品系亲缘关系较近,同属于WolbachiaB大组Con组。

Wolbachia bacteria is an obligate intracellular bacterium found in a wide range of invertebrate hosts and cause alterations in host reproductive phenotypes, including cytoplasmic incompatibility, feminization, parthenogenesis, male killing, fitness alterations, and other phenomena. Small brown planthoppers（ Laodelphax striatellus ）was the most important vector of some plant virus transmission and the populations were trended to increase in recent years which caused serious damage to rice and maize plant. Thus the infection of Wolbachia in small brown planthoppers from different regions was detected by polymerase chain reaction（PCR）using the universal primer 81F and 691Rs of the surface protein gene of Wolbachia（ wsp gene, encoding the outer membrane protein of the symbiotic bacteria）in this paper. The results demonstrated that planthoppers from Quyang county and Rongcheng county were infected with Wolbachia and the infection rate was 85.6 % and 88.4 % respectively. The PCR products were sequenced and analyzed. It was showed that a region of 601 - 603 bp of Wolbachia wsp gene were amplified from the total DNA of small brown planthopper and there was 100% nucleotide identity with other strains derived from Sogatella fureifera and Laodelphax striatellus. It indicated that the wolbachia infected in small brown planthopper from Hebei province had close relatives with the strain wFur（derived from Sogatellafureifera） and wStri （derived from Laodelphax striatellus ）. They all belong to the Con group of B supergroup. Therefore, it was possible that wolbachia infected Laodelphax striatellus derived from the same group.