Abstract
Four PCR primers that are useful to determine the nucleotide sequences of the rDNA of the powdery mildew fungi were newly designed. These primers provide both enough stability to work on a wide range of powdery mildews and enough specificity to eliminate contaminating DNA by PCR. DNA sequences of the rDNA ITS region were successfully obtained from specimens that were contaminated by other fungi. In addition, sequence results of the 18S and 28S rDNA were dramatically improved by using these primers in most of the specimens examined.
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Cunnington, J. H., Takamatsu, S., Lawrie, A. C. and Pascoe, I. G. 2001. Molecular identification of anamorphic powdery mildews (Erysiphales). Mycol. Res.105 (In press).
Hirata, T. and Takamatsu, S. 1996. Nucleotide sequence diversity of rDNA internal transcribed spacers extracted from conidia and cleistothecia of several powdery mildew fungi. Mycoscience37: 283–288.
Kusaba, M. and Tsuge, T. 1995. Phylogeny ofAlternaria fungi known to produce host-specific toxins on the basis of variation in internal transcribed spacers of ribosomal DNA. Curr. Genet.28: 491–498.
Mai, J. C. and Coleman, A. W. 1997. The internal transcribed spacer 2 exhibits a common secondary structure in green algae and flowering plants. J. Mol. Evol.44: 258–271.
Mori, Y., Sato, Y. and Takamatsu, S. 2000a. Evolutionary analysis of the powdery mildew fungi (Erysiphales) using nucleotide sequences of the nuclear ribosomal DNA. Mycologia92: 74–93.
Mori, Y., Sato, Y. and Takamatsu, S. 2000b. Molecular phylogeny and radiation time of Erysiphales inferred from the nuclear ribosomal DNA sequences. Mycoscience41: 437–447.
Müller, E. M., Bahnweg, G., Sandermann, H. and Geiger, H. H. 1992. A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues. Nucleic Acids Res.20: 6115–6116.
Schlötter, C., Hauser, M.-T., von Haeseler, A. and Tautz, D. 1994. Comparative evolutionary analysis of rDNA ITS regions inDrosophila. Mol. Biol. Evol.11: 513–522.
Takamatsu, S., Hirata, T. and Sato, Y. 1998. Phylogenetic analysis and predicted secondary structures of the rDNA internal transcribed spacers of the powdery mildew fungi (Erysiphaceae). Mycoscience39: 411–453.
Van der Sande, C. A. F. M., Kwa, M., Van Nues, R. W., Van Heerikhuizen, H., Raué, H. A. and Planta, R. J. 1992. Functional analysis of internal transcribed spacer 2 ofSaccharomyces cerevisiae ribosomal DNA. J. Mol. Biol.223: 899–910.
Van Nues, R. W., Rientjes, J. M. J., Morré, S. A., Mollee, E., Planta, R. J., Venema, J. and Raué, H. A. 1995. Evolutionarily conserved structural elements are critical for processing of internal transcribed spacer 2 fromSaccharomyces cerevisiae precursor ribosomal RNA. J. Mol. Biol.250: 24–36.
White, T. J., Bruns, T. D., Lee, S. and Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal genes for phylogenetics. In: PCR protocols: a guide to methods and applications, (ed. by Innis, M. A., Gelfand, D. H., Sninsky, J. J. and White, T. J.), pp. 315–322. Academic Press, San Diego.
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Takamatsu, S., Kano, Y. PCR primers useful for nucleotide sequencing of rDNA of the powdery mildew fungi. Mycoscience 42, 135–139 (2001). https://doi.org/10.1007/BF02463987
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DOI: https://doi.org/10.1007/BF02463987