Rapid detection of Rhizoctonia species, causal agents of rice sheath diseases, by PCR-RFLP analysis using an alkaline DNA extraction method

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Abstract

Restriction fragment length polymorphism (RFLP) analysis for DNA products amplified by the polymerase chain reaction (PCR) was used for the direct detection of Rhizoctonia solani AG 1 IA and AG 2-2 IIIB, R. oryzae, R. oryzae-sativae and R. fumigata from the diseased rice sheaths. A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract parasite DNA from diseased rice sheaths. 28S ribosomal DNA (rDNA) derived from fungal genomic DNA extracted by the alkaline method was specifically PCR-amplified. The results of PCR-RFLP analysis for DNA samples from artificially inoculated rice sheath tissues with each Rhizoctonia spp. and the corresponding culture on the medium using two restriction enzymes. HhaI and MspI, showed identical polymorphisms. PCR-RFLP analysis using DNA samples from naturally infected rice sheath tissues also revealed the possibility of direct diagnosis of R. solani AG 1 IA, R. oryzae and R. oryzae-sativae.

Original languageEnglish
Pages (from-to)451-454
Number of pages4
JournalMycoscience
Volume38
Issue number4
DOIs
Publication statusPublished - Jan 1 1997

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Rhizoctonia
extraction method
polymerase chain reaction
restriction fragment length polymorphism
polymorphism
rice
Oryza
DNA
Thanatephorus cucumeris
methodology
sodium hydroxide
ribosomal DNA
analysis
detection
hydroxide
genetic polymorphism
genomics
parasite
parasites
sampling

All Science Journal Classification (ASJC) codes

  • Ecology, Evolution, Behavior and Systematics

Cite this

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title = "Rapid detection of Rhizoctonia species, causal agents of rice sheath diseases, by PCR-RFLP analysis using an alkaline DNA extraction method",
abstract = "Restriction fragment length polymorphism (RFLP) analysis for DNA products amplified by the polymerase chain reaction (PCR) was used for the direct detection of Rhizoctonia solani AG 1 IA and AG 2-2 IIIB, R. oryzae, R. oryzae-sativae and R. fumigata from the diseased rice sheaths. A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract parasite DNA from diseased rice sheaths. 28S ribosomal DNA (rDNA) derived from fungal genomic DNA extracted by the alkaline method was specifically PCR-amplified. The results of PCR-RFLP analysis for DNA samples from artificially inoculated rice sheath tissues with each Rhizoctonia spp. and the corresponding culture on the medium using two restriction enzymes. HhaI and MspI, showed identical polymorphisms. PCR-RFLP analysis using DNA samples from naturally infected rice sheath tissues also revealed the possibility of direct diagnosis of R. solani AG 1 IA, R. oryzae and R. oryzae-sativae.",
author = "Matsumoto Masaru",
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AU - Masaru, Matsumoto

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N2 - Restriction fragment length polymorphism (RFLP) analysis for DNA products amplified by the polymerase chain reaction (PCR) was used for the direct detection of Rhizoctonia solani AG 1 IA and AG 2-2 IIIB, R. oryzae, R. oryzae-sativae and R. fumigata from the diseased rice sheaths. A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract parasite DNA from diseased rice sheaths. 28S ribosomal DNA (rDNA) derived from fungal genomic DNA extracted by the alkaline method was specifically PCR-amplified. The results of PCR-RFLP analysis for DNA samples from artificially inoculated rice sheath tissues with each Rhizoctonia spp. and the corresponding culture on the medium using two restriction enzymes. HhaI and MspI, showed identical polymorphisms. PCR-RFLP analysis using DNA samples from naturally infected rice sheath tissues also revealed the possibility of direct diagnosis of R. solani AG 1 IA, R. oryzae and R. oryzae-sativae.

AB - Restriction fragment length polymorphism (RFLP) analysis for DNA products amplified by the polymerase chain reaction (PCR) was used for the direct detection of Rhizoctonia solani AG 1 IA and AG 2-2 IIIB, R. oryzae, R. oryzae-sativae and R. fumigata from the diseased rice sheaths. A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract parasite DNA from diseased rice sheaths. 28S ribosomal DNA (rDNA) derived from fungal genomic DNA extracted by the alkaline method was specifically PCR-amplified. The results of PCR-RFLP analysis for DNA samples from artificially inoculated rice sheath tissues with each Rhizoctonia spp. and the corresponding culture on the medium using two restriction enzymes. HhaI and MspI, showed identical polymorphisms. PCR-RFLP analysis using DNA samples from naturally infected rice sheath tissues also revealed the possibility of direct diagnosis of R. solani AG 1 IA, R. oryzae and R. oryzae-sativae.

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