Molecular genetic characterization of bacterial isolates causing brown blotch on cultivated mushrooms in Japan |
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Authors: | Thorn Greg Tsuneda Akihiko |
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Affiliation: | (1) Center for Microbial Ecology, Michigan State University, 48824 East Lansing, MI, USA;(2) Tottori Mycological Institute, 211 Kokoge, 689-11 Tottori, Japan;(3) Present address: Department of Botany, University of Wyoming, 8207 Laramie, WY, USA |
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Abstract: | The polymerase chain-reaction (PCR) was used to amplify 16S ribosomal DNA (16S rDNA) from bacteria, identified asPseudomonas tolaasii orP. fluorescens, causing brown blotch on cultivated mushrooms in Japan. PCR-amplified 16S rDNA was analyzed on the basis of nucleotide sequence and restriction fragment length polymorphisms (RFLP) to determine the specific identity of isolates. Banding patterns obtained through PCR using primers corresponding to repetitive extragenic palindromic sequences of enteric bacteria (REP-PCR) were used to determine the relatedness of conspecific isolates. AllP. tolaasii isolates and a mushroom pathogen identified asP. fluorescens had identical RFLP patterns and partial 16S sequences, and are considered conspecific. An isolate ofP. fluorescens from creamery wastes (IFO 3507) differed slightly from isolates ofP. tolaasii in both 16S sequence (0.8%) and RFLP patterns (d=0.08), and had almost entirely different REP-PCR bands (d=0.88–1.0). Phylogenetic analyses based on 16S sequences indicated thatP. tolaasii andP. fluorescens are close members ofPseudomonas sensu stricto. REP-PCR shows promise in characterizing isolates pathogenic on different mushroom crops. Two isolates ofP. tolaasii pathogenic onPleurotus ostreatus had identical banding patterns, but three isolates fromLentinula edodes showed the greatest diversity. Contribution No. 312 of the Tottori Mycological Institute, Totori, Japan. |
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Keywords: | Agaricus brunnescens bacterial disease Lentinula edodes Pseudomonas fluorescens Pseudomonas tolaasii REP-PCR |
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