淡水鱼类粘孢子虫的18S rDNA分子系统学研究
PHYLOGENETIC ANALYSES OF THE MYXOSPOREANS PARASITIC IN FRESHWATER FISH OF CHINA BASED ON 18S RIBOSOMAL DNA
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摘要: 利用两个通用引物myxoF('-CGCGGTAATTCCAGCTCCAGTAG-3')和myxoR('-ACCAGGTAAGTTTTCCCGTGTTGA-3')成功扩增出圆形碘泡虫、全圆碘泡虫、武汉单极虫、微山尾孢虫和库班碘泡虫种粘孢子虫的部分18S rDNA序列,其GenBank登录号为:AY16179-AY16183.并结合GenBank其他13个相关序列构建了18个物种的分子系统树.结果表明,碘泡虫,尾孢虫和单极虫较Tetracapsula bryozoides和"PKX"分化晚,它们形成了2个聚类:T.wuhanensis-T.hovorkai-M.rotundatus-M.rotundus-M.bibullatus-M.pellicides-M.pendula-H.salminicola聚类和H.exilis-H.ictaluri-M.spinacurvatura-M.osburni-H.lesteri-H.weishanensis-M.kubanicum的聚类;同一属内,采自国内的种类并没有因为地域关系而形成独立的分支,而是与国外的种类交叉在一起,这意味着粘孢子虫种类之间的地域差别并不大;碘泡虫和尾孢虫在进化上都不是单系的,而且分子数据难以将这两个属的种类分开,因此尾孢虫的尾突可能并不是有效的分类依据,而是和碘泡虫的壳片突起同源的一种附属结构.Abstract: Partial 18S rDNA sequences of five myxosporean species, Myxobolus rotundus,M.rotundatus,Thelohanellus wuhanensis,Henneguya weishanensis and M.cubanicum were amplified utilizing two common primers myxoF(5′-CGCGGTAATTCCAGCTCC AGTAG-3′)and myxoR(5′-ACCAGGTAAGTTTTCCCGTGTTGA-3′)and stored in the GenBank with the access number AY165179-AY165183 By combining the 18S rDNA sequences of other 13 species available in the GenBank a parsimonious tree of 18 species was built with Polypodium hydriforme used as the outgroup.Results show Tetracapsula bryozoides and “PKX"organism lie on the root within the Myxozoa before the divergence of other major groups which form two clades as T.wuhanensis-T. hovorkai-M.rotundatus-M.rotundus-M.bibullatus-M.pellicides-M.pendula-H.salminicola and H.exilis-H. ictaluri-M.spinacurvatura-M.osburni-H.lesteri-H.weishanensis-M.kubanicum. Among the genus myxobolus, 3 species found in China did not form a clade but grouped with the European species,indicating that the geographic divergence of Myxobolus may not be distinct.Neither Myxobolus nor Henneguya was monophyletic and these two genera can not be distinguished using the molecular data.It is further proposed that the caudal protrusion of Henneguya may not be a valid taxonomic character but an appending structure homologous with the “V"projections in Myxobolus.
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Keywords:
- Myxosporean /
- phylogenetic tree /
- 18S ribosomal DNA /
- Myxobolus /
- Henneguya /
- Thelohanellus
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D,Smith,et al.Molecular evidence that the myxozoan protists are metazoans[J].Science,1994,265:1719-1721[5] Walliker L.The nature of the iodinophilous vacuole of myxosporidian spores and a proposal to synonymize the genus Myxosoma Thelohan,1892 with the genus Myxobolus Butschli,1882[J].J Protozool,1968,15:571-575[6] Lom J.On a new taxonomic character in Myxosporidia,as demonstrated in description of two new species of Myxobolus[J].Folia Parasitol,1969,16:97-103[7] Desser S S,Paterson W B.Ultrastructural and cytochemical observations on sporogenesis of Myxobolus sp.(Myxosporida:Myxobolidae)from the common shiner Notropis cornutus[J].J Protozool,1978,25:314-326[8] Lom J.Hoferellus cyprini(Doflein,1898)Berg,1898(syn:Mitraspora cyprini Fujita,1912),Myxobilatus nostalgicussp.c.and related species:partial revision of two Myxosporean genera[J].Folia Parasitol,1986,33:289-296[9] Lom J.Hoferellus cyprini Doflein,1898 from carp kidney:a well established Myxosporean species or a sequence in the developmental cycle of Sphaerospora renicola Dykova and Lom,1982[J]?Protistol,1985,2:195-206[10] Andree K B,Gresoviac S J,Hedrick R P.Small subunit ribosomal RNA sequences unite alternate actinosporean and myxosporean stages of Myxobolus cerebralis,the causative agent of whirling disease in salmonid fish[J].J Euk Microbiol,1997,44:208-215[11] Anderson C L,Canning E U,Schafer S M,et al.Molecular confirmation of the life cycle of Thelohanellus hovorkai Achmerov,1960(Myxozoa:Myxosporea)[J].Bull Eur Ass Fish Pathol,2000,20:111-114[12] Andree K B,El-Matbouli M,Hoffman R W,et al.Comparison of 18S and ITS-1 sequences of selected geographic isolates of Myxobolus cerebralis[J].Int J Parasitol,1999,29:771-775[13] Andree K B,Szekely C,Molnar K,et al.Relationships among members of the genus Myxobolus(Myxozoa:Bivalvulida)based on small subunit ribosomal DNA sequences[J].J Parasitol,1999,85:68-74[14] Chen Q L,Ma C L.Myxozoa,Myxosporea.Fauna sinica[M].Beijing:Science Press.1998[陈启鎏,马成伦.中国动物志,粘体动物门,粘孢子纲.北京:科学出版社.1998][15] Kent M L,Andree K B,Bartholomew J L,et al.Recent advances in our knowledge of the Myxozoa[J].J Euk Microbiol,2001,48:395-413[16] Sambrook J,Fritsch E F,Maniatis T.Molecular cloning:a laboratory manual,2nd ed[M].New York:Cold Spring Harbor Laboratory Press.1989[17] DNASTAR incorporation.Windows 32 Primerselect,Version 4.01[P].Madison,Wisconsin.1999[18] Thompson J D,Gibson T J,Plewniak F,et al.The ClustalX windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nuc Acids Res,1997,24:4876-4882[19] Felsenstein J.PHYLIP-Phylogeny Inference Package,Version 3.6(alpha3)[P],Seattle,Washington:Department of Genome Sciences,University of Washington.2002[20] Anderson C L,Canning E U,Okamura B.18S rDNA sequences indicate that "PKX"organism parasitizes Bryozoa[J].Bull Eur Ass Fish Pathol.1999,19:94-97[21] Anderson C L,Canning E U,Okamura B.Molecular data implicate bryozoans as host for "PKX"(Phylum Myxozoa)and identify a clade of bryozoan parasites within the Myxozoa[J].Parasitology.1999,119:556-561[22] Canning E U,Okamura B,Curry A.Development of a myxozoan parasite Tetracapsula bryozoides gen.c.et sp.n.in Cristatella mucedo(Bryozoa:Phylactolaemata)[J].Folia Parasitol,1996,43:249-261
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