扶桑绵粉蚧共生细菌 Delftia 的多样性与侵染动态  

Diversity and infection dynamics of Delftia in the cotton mealybug,Phenacoccus solenopsis(Hemiptera:Pseudococcidae)

作　　者：郑奕然 倪美虹 杨欣怡 李紫成 蒋明星[1] ZHENG Yi-Ran;NI Mei-Hong;YANG Xin-Yi;LI Zi-Cheng;JIANG Ming-Xing(Key Laboratory of Agricultural Entomology,Ministry of Agriculture,Key Laboratory of Biology of Crop Pathogens and Insects,Zhejiang Province,Institute of Insect Sciences,Zhengjiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学昆虫科学研究所,农业部农业昆虫学重点实验室,浙江省作物病虫生物学重点实验室,杭州310058

出　　处：《昆虫学报》2024年第7期932-942,共11页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(32072489)。

摘　　要：【目的】本研究旨在明确昆虫共生细菌伯克氏菌目(Burkholderiales)丛毛单胞菌科(Comamonadaceae)Delftia在扶桑绵粉蚧Phenacoccus solenopsis中的多样性、丰度动态和感染部位。【方法】克隆并测序扶桑绵粉蚧5个不同地理种群[浙江兰溪(寄主:木芙蓉Hibiscus mutabilis)、浙江东阳(寄主:辣椒Capsicum annuum)、浙江杭州西湖区(寄主:大花马齿苋Portulaca grandiflora)、浙江杭州临平区(寄主:大花马齿苋)和广西防城港(寄主:朱槿Hibiscus rosa-sinensis)]3龄若虫肠道中细菌的16S rRNA基因片段,对Delftia株系进行鉴定和系统发育分析;采用qPCR定量分别取食棉花和番茄的扶桑绵粉蚧不同发育阶段(1-3龄若虫、初羽化成虫及2,4和6日龄成虫以及开始产蚧后2和4 d的成虫)整虫和其中取食番茄的上述各发育阶段成虫肠道和2,4和6日龄成虫卵巢中Delftia 16S rRNA基因的拷贝数;借助荧光原位杂交技术(fluorescence in situ hybridization,FISH)检测扶桑绵粉蚧初羽化成虫肠道和马氏管中Delftia的侵染动态;分析扶桑绵粉蚧从寄主植物获得Delftia的可能性。【结果】扶桑绵粉蚧各地理种群3龄若虫所感染细菌Delftia的16S rRNA基因片段序列极为相似(相似度>99%),为同一个种Delftia sp.的不同菌株,且与部分其他昆虫、植物中及根际的Delftia系统发育关系十分接近。在若虫期,取食棉花和番茄扶桑绵粉蚧Delftia丰度均随龄期增大逐渐上升,但成虫期Delftia丰度变化因寄主植物而异,取食棉花扶桑绵粉蚧2日龄成虫内Delftia丰度较高6日龄成虫内Delftia丰度明显下降,开始产蚧后4 d成虫内Delftia丰度也有下降趋势;取食番茄扶桑绵粉蚧2日龄成虫内Delftia丰度显著下降,6日龄成虫内Delftia丰度明显上升,开始产蚧后4 d成虫内Delftia丰度也上升。初羽化成虫肠道和卵巢中Delftia丰度与扶桑绵粉蚧发育期关系密切,其中肠道中丰度随成虫发育推进呈缓慢下降趋势,卵巢中【Aim】The aim of our study is to determine the diversity,abundance dynamics and infection site of the symbiotic bacterium Delftia(Burkholderiales:Comamonadaceae)in the cotton mealybug,Phenacoccus solenopsis.【Methods】By cloning and sequencing the bacterial 16S rRNA gene fragment,Delftia strains in the gut of the 3rd instar nymphs of five geographical populations[Lanxi of Zhejiang(host:Hibiscus mutabilis),Dongyang of Zhejiang(host:Capsicum annuum),Xihu District,Hangzhou of Zhejiang(host:Portulaca grandiflora),Linping District,Hangzhou of Zhejiang(host:P.grandiflora)and Fangchenggang of Guangxi(host:Hibiscus rosa-sinensis)]of P.solenopsis were identified and phylogenetically analyzed.The copy number of Delftia 16S rRNA gene of Delftia in the whole body of P.solenopsis at different developmental stages(1st-3rd instar nymphs,newly emerged adult,and 2-,4-and 6-day-old adults,and adults at 2 and 4 d after beginning to produce nymphs)fed on cotton and tomato,and in the gut of adults at the above developmental stages and in the ovary of the 2-,4-and 6-day-old adults fed on tomato were quantified using qPCR.Fluorescence in situ hybridization(FISH)was taken to detect the infection dynamics of Delftia in the gut and Malpighian tubules of the newly emerged adult of P.solenopsis.The possibility that P.solenopsis obtained Delftia from host plants was analyzed.【Results】The bacteria of Delftia from the 3rd instar nymphs of various geographical populations of P.solenopsis had quite similar 16S rRNA gene sequences(similarity>99%),being different strains of the same Delftia species,and they had close phylogenetic relationships with those found in some of other insects,plants and rhizosphere.During the nymphal stage,the abundance of Delftia in P.solenopsis fed on cotton and tomato increased gradually with instars.During the adult stage,however,the abundance of Delftia varied with the types of host plants.The abundance of Delftia in the 2-day-old adult of P.solenopsis fed on cotton was high,that in the 6-day-old adult fed o

关 键 词：扶桑绵粉蚧 共生细菌 Delftia 细菌多样性 细菌丰度 侵染动态 

分 类 号：S433.3[农业科学—农业昆虫与害虫防治]