title: Quantification of rice blast disease progressions through Taqman real-time PCR.
abstract: Rice blast caused by Magnaporthe oryzae is a major disease in the paddy field and also a representative model system in the investigation of plant-microbe interactions. This study was undertaken to provide the quantitative evaluation method that specifically determines the amount of M. oryzae proliferation in planta. Real-time PCR was used as the detection strategy in combination with the primer pair and Taqman probe specific to MHP1, a unigene encoding HYDROPHOBIN that is indispensable for normal virulence expression. Based on the crossing point values from the PCR reactions containing a series of increasing concentration of cloned amplicon or fungal genomic DNA, correlation among the template's copy number or its amount and amplification pattern was calculated. Reliability of this equation was further confirmed using the DNA samples from the rice leaves infected with compatible or incompatible strains of M. oryzae. The primer pair used in the Taqman real-time PCR reaction can recognize the existence of fungal DNA as low as 1 pg. In sum, our quantitative evaluation system is applicable and reliable in the blast diagnosis and also in the estimation of objective blast disease progression.
authors: Rural Development Administration, National Academy of Agricultural Science, Suwon 441-707, South Korea.
authors: Mukhamad
authors: M
authors: Su'udi
authors: Jinyeong
authors: J
authors: Kim
authors: Jong-Mi
authors: JM
authors: Park
authors: Shin-Chul
authors: SC
authors: Bae
authors: Donghern
authors: D
authors: Kim
authors: Yong-Hwan
authors: YH
authors: Kim
authors: Il-Pyung
authors: IP
authors: Ahn
mesh_terms: D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005656:Fungal Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D018628:Gene Dosage; D020082:Magnaporthe / Q000254:growth & development* / Q000472:pathogenicity; D012275:Oryza / Q000382:microbiology*; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction; D013997:Time Factors; D037521:Virulence Factors / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D005656:Fungal Proteins; D037521:Virulence Factors
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