Abstract
We examined the usefulness of primer sets designed to amplify introns within conserved genes in filamentous ascomycetes to differentiate 35 isolates representing six different species of Fusarium commonly found in association with conifer seedlings. We analyzed restriction fragment length polymorphisms (RFLP) in five amplified PCR products from each Fusarium isolate. The primers used in this study were constructed on the basis of sequence information from the H3, H4, and (beta)-tubulin genes in Neurospora crassa. Primers previously developed for the intergenic transcribed spacer region of the ribosomal DNA were also used. The degree of interspecific polymorphism observed in the PCR products from the six Fusarium species allowed differentiation by a limited number of amplifications and restriction endonuclease digestions. The level of intraspecific RFLP variation in the five PCR products was low in both Fusarium proliferatum and F. avenaceum but was high in a population sample of F. oxysporum isolates. Clustering of the 35 isolates by statistical analyses gave similar dendrograms for H3, H4, and (beta)-tubulin RFLP analysis, but a dendrogram produced by intergenic transcribed spacer analysis varied in the placement of some F. oxysporum isolates.
Full Text
The Full Text of this article is available as a PDF (275.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Crowhurst R. N., Hawthorne B. T., Rikkerink E. H., Templeton M. D. Differentiation of Fusarium solani f. sp. cucurbitae races 1 and 2 by random amplification of polymorphic DNA. Curr Genet. 1991 Nov;20(5):391–396. doi: 10.1007/BF00317067. [DOI] [PubMed] [Google Scholar]
- Glass N. L., Donaldson G. C. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl Environ Microbiol. 1995 Apr;61(4):1323–1330. doi: 10.1128/aem.61.4.1323-1330.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glass N. L., Smith M. L. Structure and function of a mating-type gene from the homothallic species Neurospora africana. Mol Gen Genet. 1994 Aug 15;244(4):401–409. doi: 10.1007/BF00286692. [DOI] [PubMed] [Google Scholar]
- Gower J. C. A comparison of some methods of cluster analysis. Biometrics. 1967 Dec;23(4):623–637. [PubMed] [Google Scholar]
- Guadet J., Julien J., Lafay J. F., Brygoo Y. Phylogeny of some Fusarium species, as determined by large-subunit rRNA sequence comparison. Mol Biol Evol. 1989 May;6(3):227–242. doi: 10.1093/oxfordjournals.molbev.a040548. [DOI] [PubMed] [Google Scholar]
- Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
- O'Donnell K. Ribosomal DNA internal transcribed spacers are highly divergent in the phytopathogenic ascomycete Fusarium sambucinum (Gibberella pulicaris). Curr Genet. 1992 Sep;22(3):213–220. doi: 10.1007/BF00351728. [DOI] [PubMed] [Google Scholar]
- Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Zyl-Schalekamp C. Self-medication in three Orange Free State communities. S Afr Med J. 1993 May;83(5):345–346. [PubMed] [Google Scholar]