Conflicting results obtained by RAPD-PCR and large-subunit rDNA sequences in determining and comparing yeast strains isolated from flowers: a comparison of two methods.
Sixty-six yeast strains isolated from the nectar of various plant species in Central Europe were characterized by randomly amplified polymorphic DNA PCR (RAPD-PCR) and by sequencing of the variable D1/D2 domain of large-subunit (26S) rDNA. The usefulness of both methods for the determination and comparison of unknown ascomycetous and basidiomycetous yeast strains was compared and evaluated. The reproducibility of RAPD-PCR was shown to be low and the information obtained by this method was clearly not as precise as that obtained from sequence analysis. Numerous imponderables make RAPD-PCR analysis unreliable, at least as a means of identifying yeasts in ecological studies. The lack of standard protocols for RAPD-PCR analysis and the absence of a general database of banding patterns made it impossible to identify unknown yeast strains or to recognize new species. In contrast to RAPD-PCR, sequence analysis of the D1/D2 domain was found to be a fast and reliable method for the rapid identification of yeast species and was also shown to be an invaluable tool for the discovery of new species.
Herzberg
Michael
M
Fischer
Reinhard
R
Titze
Andreas
A
D001203:Ascomycota / Q000145:classification* / Q000235:genetics; D001487:Basidiomycota / Q000145:classification* / Q000235:genetics; D004275:DNA, Ribosomal / Q000032:analysis; D035264:Flowers / Q000382:microbiology*; D019684:Magnoliopsida / Q000382:microbiology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012335:RNA, Ribosomal / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA
D004275:DNA, Ribosomal; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S