水霉菌的形态及ITS区分子鉴定  

SAPROLEGNIA IDENTIFICATION BASED ON THEIR MORPHOLOGICAL CHARACTERISTICS AND ITS rDNA REGION

作　　者：可小丽[1,2] 汪建国[1] 顾泽茂[3] 李明[4] 龚小宁[1] 

机构地区：[1]中国科学院水生生物研究所,武汉430072 [2]中国科学院研究生院,北京100049 [3]华中农业大学水产学院,武汉430070 [4]武汉工业学院饲料科学系,武汉430023

出　　处：《水生生物学报》2010年第2期293-301,共9页Acta Hydrobiologica Sinica

基　　金：湖北省科技攻关计划项目(2007AA203A01);973计划项目(2009CB118700);中国科学院领域前沿项目(053903)资助

摘　　要：研究利用形态学和分子生物学相结合的方法,对从患病黄颡鱼病灶处分离的水霉菌株(HSY)的分类地位进行了研究。结果显示,该菌在18-23℃均能形成游动孢子囊,但只在20℃才大量释放游动孢子;在15℃培养约3周出现大量的藏卵器,多数顶生,形态与Saprolegnia litoralis和Saprolegnia ferax的均非常相似。序列比对分析显示,HSY菌株与S.litoralis、S.bulbosa、S.oliviae、S.longicaulis、S.ferax、S.mixta和S.anomalies的ITS区(包括5.8S rDNA)序列相似性均高于99%。系统发育分析显示,所选的真菌分为3个群,HSY落于第一个群,MP树显示HSY菌株与S.litoralis、S.bulbosa、S.oliviae、S.longicaulis、S.ferax以及S.mixta枝系均互相平行,而NJ树显示其与S.ferax和S.mixta更近。综合形态学、ITS区序列及系统发育分析,将HSY菌株暂定为多子水霉(Saprolegnia ferax)。Saprolegnia is one of the main genus of water molds responsible for fungal infections of freshwater fish and their eggs. These infections are usually termed ＂saprolegniasis＂, which can cause severe losses of freshwater fish in both nature and commercial fish farms. However, identification of the causative agent is often unmanageable, especially to those isolated from fish lesions. Traditionally, the generic definition was mainly based on the asexual characters, espe-cially the mode of zoospores discharge, and species differentiation was mainly on the features of sexual reproductive organs including oogonia, antheridia, antheridial origin and oospore. However, many of these characters may be variable in one species or similar in different species and some strains either lose the ability to produce sexual reproduction or cannot be induced to form them under laboratory conditions. So, it is difficult to make definitive identification only using traditional morphological criteria in species level. In this study, we identified one saprolegnia isolate HSY obtained from yellow catfish （Pelteobagrus fulvidraco） based on the morphological and molecular characteristics. Results showed Isolate HSY grew luxuriantly on hemp seeds in water at 18—23 ℃ and were measured 1.5—4.0cm in diameter. The hyphae were stout, sparingly branched and measured between 20—58μm in diameter. Zoosporangia had formed abundantly and were often fusiform, clavate, straight or bent, lateral or terminal and renewed internally, measured 110—490 × 25—62μm. Primary zoospore dis-charge was typically saprolegnoid, pyriform; secondary zoospores emerged from the encysted, reniform; encysted zoo-spores were globose and measured 9—12μm in diameter. Oogonia were formed abundantly within three weeks at 15℃, not 18 ℃ and 23℃, mainly terminal, rarely lateral, and measured 75—108μm in diameter. Oogonial walls were sooth, and stalks were stout and bent. The attachment of antheridial cells was moderate （mainly androgynous and monoc

关 键 词：卵菌纲 水霉 分类 ITSrDNA 系统发育 

分 类 号：S941.43[农业科学—水产养殖]