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Microbiology and Biotechnology Letters

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Microbiol. Biotechnol. Lett. 2009; 37(1): 1-9

Multiplex PCR을 이용한 생물방제균 Bacillus subtilis AH18의 토양내 Genetic Monitoring

Genetic Monitoring of Plant Growth Promoting Rhizobacterium (PGPR), Bacillus subtilis AH18 using Multiplex PCR in Field Soil

Sang-Min Woo , Jong-Hui Lim , Hee-Young Jeong and Sang-Dal Kim

Department of Applied Microbiology, Yeungnam University, 1Department of Applied Microbiology, Yeungnam University, 2Department of Applied Microbiology, Yeungnam University, 3Department of Applied Microbiology, Yeungnam University

Abstract

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The genetic monitoring method was developed for the rapid detection of the PGPR and biocontrol agent, B. subtilis AH18 in red-pepper field soil by multiplex PCR using sid, aec and cel gene primers. The monitoring of B. subtilis AH18 in the soil was carried by amplified a 2,3-dihydro-2,3-dihydroxy benzoate dehydrogenase [EC: 1. 3. 1. 28]gene (sid - 794 bp : EF408238) which is a key enzyme of siderophore synthesis, an auxin efflux carrier gene (aec - 1,052 bp : EF408239) and a cellulase gene (cel - 1,582 bp : EF070194). The natural un sterilized soil was inoculated with B. subtilis AH18 to determine the sensitivity ($1.8 imes10^5$ cfu/g) of multiplex PCR for the rapid dectection and then the strain was monitored successfully in rhizosphere or non-rhizosphere soil of red-pepper cultural soil. At 3 weeks after the treatment, density of the strain was monitored more abundantly in rhizosphere soil.

Keywords: Genetic monitoring, multiplex PCR (polymerase chain reaction), Bacillus subtilis AH18, PGPR, biocontrol

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