象山港养殖大黄鱼寄生新贝尼登虫成虫形态学和28S rDNA,ITS1分子鉴定
28S rDNA, ITS1 and adults morphological identification of Neobenedenia in cultured Pseudosciaena crocea at Xiangshan Harbor
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摘要: 贝尼登类单殖吸虫是象山港海水养殖鱼类中一类危害严重的寄生虫。应用PCR扩增及DNA序列分析的分子生物学手段,并结合对成虫的形态学分析,对象山港养殖大黄鱼体表寄生的贝尼登类单殖吸虫(记作XSp)进行了种类鉴定,结果表明XSp从形态特征上属于新贝尼登虫属,与梅氏新贝尼登虫高度相似。扩增得到XSp的28S rDNA和ITS1序列长度分别为393和427 bp,与梅氏新贝尼登虫和鱾新贝尼登虫的5个28S rDNA 序列、2个ITS1序列的比对分析显示相似性除1个为97.4%外其余均大于99%,提示XSp与这几个鱾新贝尼登虫和梅氏新贝尼登虫为种内关系,而XSp与贝尼登虫的3个28S rDNA 和1个ITS1序列相似性分别为84.3%~89.5%和60.2%。系统进化树显示该吸虫与梅氏新贝尼登虫和鱾新贝尼登虫形成一个紧密的簇,而与贝尼登虫亲缘关系较远。根据普通生物学和序列特征分析,将XSp定种为梅氏新贝尼登虫,并且支持Whittington和Horton (1996)提出的梅氏新贝尼登虫和鱾新贝尼登虫为同种异名的分类观点。Abstract: In recent years, Neobenedenia has caused serious damage on cage-cultured fish at Xiangshan Harbor, Zhejiang Province, China. The Neobenedenia adults (denoted by XSp) were collected from cultured Pseudosciaena crocea. The XSp sequences of 28S coding region of rRNA gene (28S rDNA) and the first internal transcribed spacer (ITS1) as well as morphology were studied for identification. The morphological characteristics showed that XSp belongs to Neobenedenia which was morphologically similar to Neobenedenia melleni. In XSp, the length of 28S rDNA was 393 bp and ITS1 was 427 bp. In phylogenetic analysis, Neobenedenia and Benedenia were split into two clades, and the XSp was in clade Neobenedenia. The sequences identities of 28S rDNA and ITS1 were 84.3%~89.5% and 60.2% among XSp and Benedenia, respectively. With one exception of 97.4%, the sequences identiteis of 28S rDNA and ITS1 among XSp, N. melleni and N. girellae were above 99%, which indicated that they are the same species. The results of two identification methods showed that the Neobenedenia at Xiangshan Harbor was N. melleni, and supported the opinion in which Whittington & Horton (1996) proposed that N. melleni and N. girellae were synonymy.
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Key words:
- Capsalidae /
- Neobenedenia melleni /
- morphological characteristics /
- 28S rDNA /
- ITS1 /
- taxonomy
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杨文川,李立伟,石磊,等. 福建海水养殖鱼类寄生贝尼登虫病原学研究[J]. 台湾海峡,2001,20(2):205-209. WHITTINGTON I D,HORTON M A. A revision of Neobenedenia Yamaguti,1963 (Monogenea:Capsalidae) including a redescription of N. melleni (MacCallum,1927) Yamaguti,1963[J]. Journal of Natural History,1996,30:1113-1156. YAMAGUTI S. System Helminthum:Vol.Ⅳ. Monogenea and Aspidocotylea[M]. New York:John Wiley and Dons Interscience Publishers,1963. PERKINS E M,DONNELLAN S C,BERTOZZI T,et al. Looks can deceive:molecular phylogeny of a family of flatworm ectoparasites (Monogenea:Capsalidae) does not reflect current morphological classication[J]. Molecular Phylogenetics and Evolution,2009,52:705-714. PERKINS E. Family ties:molecular phylogenetics,evolution and radiation of flatworm parasites (Monogenea:Capsalidae) . South Australia:The University of Adelaide,2010. MOLLARET I,JAMIESON B G M,ADLARD R D,et al. Phylogenetic analysis of the Monogenea and their relationships with Digenea and Eucestoda inferred from 28S rDNA sequences[J]. Molecular and Biochemical Parasitology,1997,90(2):433-438. LITTLEWOOD T J,JOHNSTON D A. Molecular phylogenetics of the four schistosoma species groups determined with partial 28S ribosomal RNA gene sequences[J]. Parasitology,1995,111(02):167-175. MORGAN J A T,BLAIR D. Nuclear rDNA ITS sequence variation in the trematode genus Echinostoma:an aid to establishing relationships within the 37-collar-spine group[J]. Parasitology,1995,111(05):609-615. VAN HERWERDEN L,BLAIR D,AGATSUMA T. Intra- and inter-specific variation in nuclear ribosomal internal transcribed spacer 1 of the Schistosoma japonicum species complex[J]. Parasitology,1998,116(04):311-317. HOSTE H,CHILTON N B,BEVERIDGE I,et al. A comparison of the first internal transcribed spacer of ribosomal DNA in seven species of Trichostrongylus (Nematoda:Trichostrongylidae)[J]. International Journal for Parasitology,1998,28(8):1251-1260. OLSON P D,LITTLEWOOD D T J. Phylogenetics of the Monogenea-evidence from a medley of molecules[J]. International Journal for Parasitology,2002,32(3):233-244. HASSOUNA N,MICHOT B,BACHELLERIE J P. The complete nucleotide sequence of mouse 28S rRNA gene:implication for the process of size increase of the large subunit rRNA in higher eukaryotes[J]. Nucleic Acids Res,1984,12:3563-3583. LI A X,WU X Y,DING X J,et al. PCR-SSCP as a molecular tool for the identification of Benedeniinae (Monogenea:Capsalidae) from marine fish[J]. Molecular and Cellular Probes,2005,19(1):35-39. TAMURA K,DUDLEY J,NEI M,et al. MEGA4:Molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Mol Bio Evol,2007,24(8):1596-1599. 张纹,王军,苏永全,等.两种新贝尼登虫28S rRNA序列分析及其亲缘关系探讨[J]. 海洋学报,2004,26(5):116-122. BONDADREANTASO M G,OGAWA K,FUKUDOME M. Reproduction and growth of Neobenedenia girellae (Monogenea,Capsalidae):a skin parasite of cultured marine fishes of Japan[J]. Fish Pathology,1995,30(3):227-231. YOSHINAGE T,NAGAKURA T,OGAWA K,et al. Attachmeng-inducing capaties of fish tissue extracts on oncomiracidia of Neobenedenia girellae (Monogenea,Capsalidae)[J]. Journal of Parasitology,2000,86(2):214-219. DEVENEY M R,CHISHOLM L A,WHITTINGTON I D. First published record of the pathogenic monogenean parasite Neobenedenia melleni (Capsalidae) from Australia[J]. Dis Aquat Org,2001,46:79-82. 丁雪娟,李安兴,张剑英,等. 海水鱼类寄生本尼登虫的ITS1序列[J]. 华南师范大学学报:自然科学版,2003,2:85-90. WHITTINGTON I D,DEVENEY M R,MORGAN J A T,et al. A preliminary phylogenetic analysis of the Capsalidae (Platyhelminthes:Monogenea:Monopisthocotylea) inferred from large subunit rDNA sequences[J]. Parasitology,2004,128(5):511-519. 杨文川,李立伟,石磊,等. 魢新本尼登虫(单殖目:多室科)的发育[J].动物学报,2002,48(1):75-79.
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