PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp.

  • 발행 : 2000.03.01

초록

To investigate the genetic relationship among 12 species belonging to the Fusarium section Martiella, Dlaminia, Gibbosum, Arthrosporiella, Liseola and Elegans, the internal transcribed spacer(ITS) regions of ribosomal DNA (rDNA) were amplified with primer pITS1 and pITS4 using the polymerase chain reaction(PCR). After the amplified products were digested with 7 restriction enzymes, restriction fragment length polymorphism (RFLP) patterns were analyzed. The partial nucleotide sequences of the ITS region were determined and compared. Little variation was observed in the size of the amplified product having sizes of 550bp or 570bp. Based on the RFLP analysis, the 12 species studied were divided into 5 RFLP types. In particular, strains belonging to the section Martiella were separated into three RFLP types. Interestingly, the RFLP type of F. solani f. sp. piperis was identical with that of isolates belonging to the section Elegans. In the dendrogram derived from RFLP analysis of the ITS region, the Fusarium spp. examined were divided into two major groups. In general, section Martiella excluding F. solani f. sp. piperis showed relatively low similarity with the other section. The dendrogram based on the sequencing analysis of the ITS2 region also gave the same results as that of the RFLP analysis. As expected, 5.8S, a coding region, was highly conserved, whereas the ITS2 region was more variable and informative. The difference in the ITS2 region between the length of F. solani and its formae speciales excluding F. solani f. sp. piperis and that of other species was caused by the insertion/deletion of nucleotides in positions 143-148 and 179-192.

키워드

참고문헌

  1. Ann.Rev.Phytopathol. v.13 The present status of Fusarium Taxonomy Booth,C.
  2. Ann. Rev. Ecol. Syst. v.22 Fungal molecular systematics Bruns, T.D.;T.J. White;J.W. Taylor
  3. Mycol. Res. v.102 Variation in the ITS and IGS regions of ribosomal DNA among the biological species of Eurpean Armillaria Chillali, M.;H. Ldder-Ighili;J.J. Guillaumin;C. Mohammed;B. Lung Escarmant;B. Botton
  4. Mycopathol. v.140 Ribosomal variation in six species of Fusarium Duggal,A.;M.T.Dumas;R.S.Jeng;M.Hubbes
  5. Mol.Biol.Evol. v.6 Phylogeny of some Fusarium species, as determined by large-subunit rRNA sequence comparison Gaudet,J.;J.Julien;J.F.Lafay;Y.Brygoo
  6. Phytopathol. v.82 Restriction fragment length polymorphism groups and physical map of mitochondrial DNA from Fusarium oxysporum f. sp. niveum Kim,D.H.;R.D.Martyn;C.W.Magill
  7. Phytophathol. v.87 Genetic diversity in the plant-pathogenci Fungus Fusarium oxysporum Kistler,H.C.
  8. Jpn.J.Genet. v.70 Restriction fragment length polymorphisms of mitochondrial DNAs from seven Fusarium species causing fusarium head blight Kodo,N.;C.Nakamura;H.Kato;T.Yoshizawa;N.Mori;C.Kaneda
  9. K.J.Mycol. v.22 Isozyme patterns of section Elegans, section Liseola and similar species in the genus Fusarium Kweon,O.Y.;B.R.Min
  10. Grzyby(Mycota), tomⅩⅩⅡ. Sierpik(Fusarium). Kwasna,H.;J.Chelkowski;P.Zajkowski
  11. Phytopathology v.63 Use of morphology and mating populations in the identification of formae speciales in Fusarium solani Matuo, T.;W.C. Synder
  12. Plant Pathology v.43 Restriction fragment length polymorphisms, races and vegetative compatibility groups within a worldwide collection of Fusarium oxysporum f. sp. gladioli Mes, J.J.;J. Van Doorn;E.J. A Roebroeck;E. Van Egmond;J, Van Artrijk;P.M. Boonekamp
  13. Kor.J.Microbiol. v.33 Comparison of Electrophoretic Karyotypes in Fusarium Min,B.R.
  14. Kor. J. Microbiol. v.36 Electrophoretic karyotypes of Fusarium oxysporum formae speciales Min, B.R.;K.A. Kim;Y.K. Choi
  15. Kor.J.Mycol. v.26 Electrophoretic Karyotyping by PFGE in the Genus Fusarium Min,B.R.;J.S.Jung;Y.K.Choi
  16. Mycoscience v.39 Differentiation of Melampsora rust species on willows in Japan using PCR-RFLP analysis of ITS regions of ribosomal DNA Nakamura H.;S. Kaneko;Y. Yamaoka;M. Kakishima
  17. Curr.Genet. v.22 Ribosomal DNA internal transcribed spacers are highly divergent in the phytopathogenic ascomycetes Fusarium sambucinum (Gibberella pulicaris) O'Donnell,K.
  18. MPMI v.8 Phylogenetic relationships of the soybean sudden death syndrome pathogen Fusarium solani f. sp. phaseoli inferred from rDNA sequence data and PCR primers for its identification O'Donnell, K.;L.E. Gray
  19. Mycologia v.90 Molecular systematics and phylogeography of the Gibberella fujikuroi species complex O'Donnell,K.;E.Cigelnik;H.I.Nirenberg
  20. Studies on the intraspecific groups in Fusarium solani(Ⅰ) On mating populations and morphologic groups in the species v.10 Sakurai, Y.;T. Matuo
  21. Ann.Rev.Phytopathol. v.33 The impact of molecular characters on systematics of filamentous ascomycetes Samuels,G.J.;K.A.Seifert
  22. Numerical taxonomy Sneath,P.H.;R.R.Sokal
  23. J.Microbiol.Meth. v.12 Grouping Fusarium Section Discolor isolates by statistical analysis of quantitative high performance liquid chromatographic data on secondary metabolite production Thrane,U.
  24. Mycologia v.88 Discordant groupings of Fusarium spp. from sections Elegans, Liseola and Dlaminia based on ribosomal ITS1 and ITS2 sequences Waalwijk,C.;R.P.Baayena;W.Gams
  25. PCR protocols Amplification and direct sequencing of fungal ribosomal genes form phylogenetics White,T.J.;T.D.Bruns;S.Lee;J.Taylor;Innis,M.A(ed.);Gelfrand,D.H.(ed.);Sninsky,J.J.(ed.);;White,T.J.(ed.)
  26. Zentralbl Bakteriol parasitendkd. Infdektionskr Ⅱ v.106 Fusarium Monographie Ⅱ. Fungi parasitici et saprophytici Wollenweber,H.W.