Detection of the pathogen of bacterial grain rot of rice from agroenvironment using membrane filtration and enrichment culture followed by PCR assay
2004
Ohara, T. (National Agricultural Research Center, Tsukuba, Ibaraki (Japan)) | Sawada, H. | Azegami, K.
ABSTRACT OHARA, T. sup(1*), SAWADA, H.2 and AZEGAMI, K.1 (2004). Detection of the pathogen of bacterial grain rot of rice from agroenvironment using membrane filtration and enrichment culture followed by PCR assay. Jpn. J. Phytopathol. 70: 115-122. The detection of the pathogen of bacterial grain rot of rice, Burkholderia glumae, from agro-environmental water using selective media is difficult, because the water generally contains a very low concentration of the targeting pathogen. A new procedure was devised using membrane filtration and enrichment culture followed by PCR. A PCR primer set, PGF1 (5'-TGTCTGACACGGAACACCTGGGTAG-3') and PPR1 (5'-AGGTTGAGTTCTCGCATTTGTGCCG-3'), was designed for specific amplification of the 16S-23SrDNA spacer region in B. glumae. The procedure made possible the detection of B. glumae from 1000 ml of irrigation water inoculated with approximately 10**0 cfu of the pathogen. We tested this procedure for various samples of agro-environmental water. B. glumae was detected from some samples of paddy water, irrigation water, river water and lake water from May through November. When the method was used on 30 species of volunteer plants in or around paddy fields with bacterial grain rot, B. glumae was detected from one plant of Monochoria vaginalis.
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