片突菱纹叶蝉感染枣疯植原体与Wolbachia的检测及系统发育分析  

Infection of Hishimonus lamellatus by Wolbachia(Rickettsiales:Alphaproteobacteria)and jujube witches’-broom phytoplasma,and phylogenetic analysis of these pathogens

作　　者：郭家洛 王金萱 贺光学 哈帕孜·恰合班 代丽珍 黄利斌 张志勇[1] 张铁强[1] 任争光[1] 王进忠[1] GUO Jia-Luo;WANG Jin-Xuan;HE Guang-Xue;Hapazi·Qiaheban;DAI Li-Zhen;HUANG Li-Bin;ZHANG Zhi-Yong;ZHANG Tie-Qiang;REN Zheng-Guang;WANG Jin-Zhong(Key Laboratory of Urban Agriculture(North China),Ministry of Agriculture and Rural Affairs,Beijing University of Agriculture,Beijing 102206,China;Forest Pest Control and Quarantine Station of Guanyang Natural Resources Bureau of Guilin City,Guangxi Province,Guilin 541600,China;Agriculture Promotion Center of Qinghe County,Xinjiang 836200,China)

机构地区：[1]北京农学院,生物与资源环境学院,农业农村部华北都市农业重点实验室,北京102206 [2]广西桂林市灌阳县自然资源局森林病虫防治检疫站,广西541600 [3]新疆青河县农业技术推广中心,新疆836200

出　　处：《应用昆虫学报》2021年第4期949-958,共10页Chinese Journal of Applied Entomology

基　　金：北京市自然科学基金与北京市教育委员会科技计划重点项目(KZ 201810020026);北京市自然科学基金(6182002);国家自然科学基金项目(31272099);北京市科学技术委员会(Z15100002115030-2)。

摘　　要：【目的】探讨片突菱纹叶蝉Hishimonus lamellatus Cai et Kuo不同种群中枣疯植原体与沃尔巴克氏体Wolbachia的感染情况和Wolbachia在不同器官组织分布,明确枣园菱纹叶蝉中Wolbachia的感染类型和分类地位,为研究Wolbachia感染对枣疯植原体潜在介体叶蝉生物学及生态学影响奠定基础。【方法】通过枣疯植原体和Wolbachia的基因特异性引物对片突菱纹叶蝉田间自然种群和实验室种群进行分子检测和鉴定。【结果】田间采集的片突菱纹叶蝉成虫植原体感染率在55%-61%之间,而Wolbachia感染率为3%-4%。田间采集的片突菱纹叶蝉自然种群经室内饲养,在1-4龄若虫中检测到Wolbachia,2-5龄若虫中检测到了植原体。片突菱纹叶蝉实验室饲养无植原体种群在其卵巢、卵和若虫中发现感染Wolbachia,在其唾液腺和消化道也检测到了Wolbachia,感染率在58%-100%之间。基于Wolbachia的wsp基因构建系统发育树,发现片突菱纹叶蝉体内的2个Wolbachia株系同属于B大组,但不同于B大组其他株系,属于新株系w Lam1和w Lam2。【结论】片突菱纹叶蝉成虫采自田间种群可以感染枣疯植原体和Wolbachia,无植原体叶蝉实验室饲养种群成虫感染Wolbachia显著高于田间种群,片突菱纹叶蝉体内2个Wolbachia株系属于B大组。这一研究结果为Wolbachia作为介体叶蝉生物防治剂进一步利用提供了基础信息。[Objectives]To explore the infection of jujube witches’-broom(JWB)phytoplasma and Wolbachia in different populations of the leafhopper,Hishimonus lamellatus Cai et Kuoh and the distribution of Wolbachia in different organs and tissues of the leafhopper,and to clarify the infection types and taxonomic status of Wolbachia in Hishimonus at jujube orchard,which laid a foundation for the study on the effects of Wolbachia infection on the biology and ecology of potential vectors of jujube witches’-broom phytoplasma.[Methods]Molecular detection and identification of field and laboratory populations of Hishimonus lamellatus were carried out by using gene-specific primers of Phytoplasma and Wolbachia.[Results]The infection rate of JWB-phytoplasma of natural populations of Hishimonus lamellatus adults collected in jujube orchards was 55%-61%,while the infection rate of Wolbachia was 3%-4%.When the natural populations of the leafhopper are reared and laid eggs,which hatched to nymphs,Wolbachia was detected in the 1st to 4th instar nymphs,and phytoplasma was detected in the 2nd to 5th instar nymphs.Wolbachia infection was found in ovaries,eggs and nymphs of the laboratory colonies reared the leafhopper phytoplasm-free,and was also detected in salivary glands and alimentary tract,with infection rates was between 58%and 100%.Phylogenetic tree was constructed based on wsp gene of Wolbachia.It was found that two Wolbachia strains belonging to group B belonged to novel strains wLam1 and wLam2,which were different from other strains of group B.[Conclusion]The natural populations of adult leafhoppers could be infected with JWB-phytoplasma and Wolbachia.The colonies of adult leafhoppers phytoplasma-free reared in the laboratory were infected with Wolbachia significantly more than those from the natural populations.Two Wolbachia strains in Hishimonus belonged to group B.The results provided basic information for the further use of Wolbachia as a biocontrol agent for leafhopper vectors.

关 键 词：片突菱纹叶蝉 潜在介体 枣疯植原体 沃尔巴克氏体 系统发育分析 

分 类 号：S436.65[农业科学—农业昆虫与害虫防治]