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Received July 11, 2013
Accepted December 13, 2013
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Quantitative analysis of the three main genera in effective microorganisms using qPCR
1Department of Biological Engineering, Inha University, Incheon 402-751, Korea 2ERC for Advanced Bioseparation Technology, Inha University, Incheon 402-751, Korea
Korean Journal of Chemical Engineering, May 2014, 31(5), 849-854(6), 10.1007/s11814-013-0274-6
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Abstract
Effective microorganism (EM) cultures have been applied in many research fields such as agriculture, environment and bioremediation. EM is a mixed culture of microorganisms including predominant populations of lactic acid bacteria and yeasts with smaller numbers of photosynthetic bacteria, actinomycetes and other types of microorganisms. Quantitative analysis of EM is requisite for the applications of EM, as its efficiency varies depending on the composition of its main genera of EM. In this study, Rhodobacter sphaeroides, Rhodopseudomonas palustris, Lactobacillus plantarum, and Saccharomyces cerevisiae, the main genera of EM were quantified by quantitative real time polymerase chain reaction, (qRT-PCR). By using selected specific primers, photosynthetic bacteria, lactic acid bacteria and yeast were quantified with high sensitivity and specificity. The ability of viable cell count by qRT-PCR was compared with agar plate cell count, showing linear relationship. Thus, PCR based quantification system is a rapid and highly specific and sensitive tool for the quantification of EM.
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References
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Deiana M, Dessi MA, Ke B, Liang YF, Higa T, Gilmour PS, Jen LS, Rahman I, Aruoma OI, Biochem. Biophys. Res. Commun., 296(5), 1148 (2002)
Herias MV, Hessle C, Telemo E, Midtvedt T, Hanson LA, Wold AE, Clin. Exp. Immunol., 116, 283 (1999)
Jin M, Wang XW, Gong TS, Gu CQ, Zhang B, Shen ZQ, Li JW, Appl. Microbiol. Biotechnol., 69(2), 229 (2005)
Ke B, Liang YF, Zhong ZX, Higa T, Aruoma OI, Environ. Toxicol. Pharmacol., 20, 313 (2005)
Mukred AM, Abd-Hamid A, Hamzah A, Yusoff WM, Pak. J. Biol. Sci., 11, 1708 (2008)
Zhou S, Wei CH, Liao CD, Wu HZ, J. Environ. Sci.(China), 20, 1514 (2008)
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Higa T, Parr JF, Beneficial and effective microorganisms for a sustainable agriculture and environment, International Nature Farming Research Center Atami, Japan (1994)
Harper-Owen R, Dymock D, Booth V, Weightman AJ, Wade WG, J. Clin. Microbiol., 37, 1469 (1999)
Valasek MA, Repa JJ, Adv. Physiol. Educ., 29, 151 (2005)
Higuchi R, Dollinger G, Walsh PS, Griffith R, Biotechnology(N Y), 10, 413 (1992)
Ririe KM, Rasmussen RP, Wittwer CT, Anal. Biochem., 245, 154 (1997)
Achenbach LA, Carey J, Madigan MT, Appl. Environ. Microb., 67, 2922 (2001)
Byun R, Nadkarni MA, Chhour KL, Martin FE, Jacques NA, Hunter N, J. Clin. Microbiol., 42, 3128 (2004)
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Herias MV, Hessle C, Telemo E, Midtvedt T, Hanson LA, Wold AE, Clin. Exp. Immunol., 116, 283 (1999)
Hierro N, Esteve-Zarzoso B, Gonzalez A, Mas A, Guillamon JM, Appl. Environ. Microbiol., 72, 7148 (2006)
