Korean Journal of Soil Science and Fertilizer (한국토양비료학회지)

Korean Society of Soil Science and Fertilizer (한국토양비료학회)
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Phenotypic Characterization of Methylotrophic N2-Fixing Bacteria Isolated from Rice (Oryza sativa L.)

벼(oryza sativa L.)에서 분리한 Methylotrophic N2-Fixing Bacteria의 형태학적 특성

  • Madhaiyan, Munusamy (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Park, Myoung-Su (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Lee, Hyoung-Seok (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Kim, Chung-Woo (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Lee, Kyu-Hoi (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Seshadri, Sundaram (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Sa, Tong-Min (Department of Agricultural Chemistry, Chungbuk National University)
  • Received : 2003.12.22
  • Accepted : 2004.02.06
  • Published : 2004.02.29
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Abstract

In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf, stem, grain, root and rhizosphere soil sainples of four rice cultivars collected from three regions of Korea. Thirty five pigmented and five non-pigmented isolates showing characteristic growth on methanul were obtained. When phylotypes were defined by performing numerical analysis of 42 characteristics, four distinct clusters were formed. While two clusters, I and IV diverged on the basis of nitrate and nitrite reduction, other two clusters, comprising only pink pigmented colonies, diverged on the basis of cellulase activity. Out of the two reference strains used in the analysis, Methyhbacterium extorquens AM1 diverged from all the clusters and M. fujisawaense KACC 10744 grouped under cluster III. All the isolates were positive for urease, oxidase, catalase and pectinase activity and negative for indole production, MR and VP test, $H_2S$ production, starch, and casein hydrolysis. No clusters were found to possess thermotolerant isolates, as no growth of the isolates was observed at $45^{\circ}C$. Two strains in cluster I were found to possess gelatin hydrolysis and methane utilizing properties respectively. Most of the isolates in all the four clusters utilized monosaccliarides, disaccharide and polyols as carbon source. Six isolates showed considerable nitrogenase activity ranging from 86.2 to $809.9nmol\;C_2H_4\;h^{-1}\;mg^{-1}$ protein.

벼(Oryza sativa L.)에 서식하고 있는 메탄올 자화세균(methylotrophic bacteria)의 군집구조를 분석하기 위하여, 국내 3지역(청원, 익산, 밀양)의 경작지 논에서 재배되고 있는 4품종(일미, 동진, 남평, 오대) 벼의 잎, 줄기, 이삭, 뿌리 및 근권토양을 수집하였다. Methanol이 유일한 탄소원으로 첨가된 선택배지를 이용하여, 분홍색 색소체를 갖는 35균주와 무색소체의 5균주를 선별하였으며, 선별균주들의 형태학적, 생리 생화학적 특성을 조사하여 4개의 군집으로 구분하였다. 군집 I 및 IV 는 각각 nitrate와 nitrite reduction 특성에 의해 구별되었으며, pink pigment colony를 형성하는 또 다른 두 개의 군집들은 cellulase 생성유무에 의하여 구분되었다. 표준균주인 Methylobacterium extorquens AM1은 분리균주들과는 다른 군집으로 구분되었으며, M. fujisawaense KACC10744는 III 군집에 속하는 것으로 분석되었다. 분리된 모든 균주들은 urease, oxidase, catalase, pectinase 활성시험에서 양성반응을 나타냈으며, indole, MR-VP, $H_2S$, starch, casein 시험에서는 음성반응을 나타내었다. 또한, 모든 분리 균주들의 열 내성은 없었으며, $45^{\circ}C$ 이상에서는 성장하지 못하였다. 군집 I에서 두개의 분리균주가 각각 gelatin 가수분해와 methane 이용능을 나타내었으며, 대부분의 균주들은 탄소원으로 monosaccharides, disaccharide, polyols를 이용하여 성장하였다. 분리 및 선별되어진 균주들 중 6 균주만이 $86.2-809.9nmol\;C_2H_4\;h^{-1}\;mg^{-1}$ protein 범위의 질소고정능을 나타내었다.

Keywords

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