灰飞虱体内一种酵母类共生菌的分子鉴定  

Identification of one yeast-like symbiont from the small brown planthopper,Laodelphax striatellus (Fallén) (Homoptera:Delphacidae)

作　　者：白旭[1] 董胜张[1] 庞琨[1] 边亚琳[1] 俞晓平[1] 

机构地区：[1]中国计量学院生命科学学院浙江省生物计量及检验检疫技术重点实验室,杭州310018

出　　处：《昆虫学报》2010年第6期640-646,共7页Acta Entomologica Sinica

基　　金：浙江省自然科学基金项目(Y3080031;Y3080410);瑞典国际科学基金IFS(C/4689-1);国家"十一五"科技支撑计划项目(2008BADA5B06-01)

摘　　要：为明确稻飞虱体内酵母类共生菌(yeast-like symbiont,YLS)的种类,采用超速离心的方法分离纯化灰飞虱Laodelphax striatellus(Falln)体内YLS,用真菌的通用引物对其18SrDNA、5.8S-ITSrDNA全长序列进行扩增。结果得到一条分子量约为2340bp的序列。序列同源性分析表明,该菌与Noda等所报道的类酵母菌的18SrDNA序列差异较大(同源性只有89.6%),而与季也蒙毕赤酵母Pichia guilliermondii有99.8%的同源性。原位杂交(ISH)和巢氏PCR均证明该菌存在于灰飞虱脂肪体和卵内,但数量较少。因此,灰飞虱体内YLS除了Noda等报道的类酵母菌外,尚存在另外一种季也蒙毕赤酵母菌。To determine the species of the yeast-like symbiont （YLS） in the rice planthoppers,the YLS was first isolated and purified by ultracentrifugation from the fat bodies of the small brown planthopper,Laodelphax striatellus （Fallén）,and then 18S rDNA and 5.8S-ITS rDNA sequences of the YLS were amplified with the general primers for fungi.Sequence homology analysis showed that 18S rDNA and 5.8S-ITS rDNA obtained were different from the strain that Noda et al.had identified previously,but showed high homology （99.8%） with those of Pichia guilliermondii.The results of in situ hybridization （ISH） and nested PCR proved that this kind of Pichia-like symbiont existed in the body of L.striatellus and involved in the transovarial transmission.The results suggest that there exist other YLS in L.striatellus besides the kind of YLS identified by Noda et al.

关 键 词：灰飞虱 类酵母共生菌 季也蒙毕赤酵母 分子鉴定 巢氏PCR 原位杂交 

分 类 号：Q965.8[生物学—昆虫学]