中国部分地区柑桔木虱体内感染Wolbachia的检测及其系统发育分析

Wolbachia是一种广泛分布于节肢动物体内的共生细菌, 该共生菌经寄主卵的细胞质传播并参与多种寄主生殖活动调控, 对节肢动物的物种进化有着重要意义并可能应用于害虫的生物防治。本研究以柑桔黄龙病（Huanglongbing, HLB）的主要传播媒介——柑桔木虱Diaphorina citri为研究对象, 通过Wolbachia的16S rDNA、 细胞分裂基因（ftsZ）、 表面蛋白基因（wsp）的特异引物对来自于中国广西北海、 广西柳州、 广东深圳、 广东阳春、 福建福州5个地区的柑桔木虱进行PCR扩增, 并对扩增产物进行克隆测序, 与GenBank中相关序列进行比对分析, 建立了柑桔木虱共生Wolbachia的系统发育树。结果显示上述5个地区的柑桔木虱均存在Wolbachia感染, 通过Wolbachia的16S rDNA, ftsZ基因和wsp基因DNA序列的系统发育分析表明, 5个地区的柑桔木虱体内的Wolbachia均属于B组; 基于wsp基因的系统发育分析进一步表明5个地区的亚洲柑桔木虱体内的Wolbachia属于B组的Con亚组, 且不同地区柑桔木虱体内的Wolbachia的16S rDNA, ftsZ基因和wsp基因序列差异不明显。研究结果为认识柑桔黄龙病传播媒介昆虫的进化及其综合防治提供了重要的参考依据。

Detection and phylogenetic analysis of Wolbachia in the Asian citrus psyllid (Diaphorina cirri) (Homoptera: Psylloidea) populations in partial areas in China

 Wolbachia is a common and widespread group of symbiotic bacteria found in arthropods, which is transmitted through the egg cytoplasm and can manipulate reproduction in their hosts in various ways. So, it has important potential significance on the research of evolution of arthropods and may be used in pest bio-control. Taking the Asian citrus psyllid (Diaphorina citri), the vector of the Huanglongbing(HLB) pathogen, as the research object, we detected the natural populations of Diaphorina citri from five areas, i.e., Beihai and Liuzhou in Guangxi, Shenzhen and Yangchun in Guangdong, and Fuzhou in Fujian, based on PCR amplification, cloning and sequencing of 16S rRNA, ftsZ and wsp genes. The obtained sequences were aligned with the published ones in database of NCBI, and the phylogenetic relationships among Wolbachia types found in D. citri and other insect species were analyzed. The results indicated that all the five populations of D. citri were infected by Wolbachia. The three gene sequences of Wolbachia and the phylogenetic tree analysis showed that Wolbachia population in D. citri belongs to supergroup B.  Further phylogenetic analysis based on the wsp gene sequences showed that the Wolbachia of D. citri belongs to Con subgroup. The 16S rRNA, ftsZ and wsp gene sequences were almost identical in the five geographic populations, respectively, and the identity is up to 99%, showing that these three gene sequences of Wolbachia in different geographic populations of D. citri had no significant difference. The results can provide useful information for understanding evolution of citrus psyllids and their integrated control.