한반도 중부지방의 벼 뿌리로부터 내생 세균의 분리와 특성 분석

Isolation and Characterization of Endophytic Bacteria from Rice Root Cultivated in Korea

  • 박수영 (한국생명공학연구원 시스템 미생물연구센터) ;
  • 양성현 (한국생명공학연구원 시스템 미생물연구센터) ;
  • 최수근 (한국생명공학연구원 시스템 미생물연구센터) ;
  • 김지현 (한국생명공학연구원 시스템 미생물연구센터) ;
  • 김종국 (경북대학교 미생물학과) ;
  • 박승환 (한국생명공학연구원 시스템 미생물연구센터)
  • Park, Soo-Young (Laboratory of Microbial Genomics, Systems Microbiology Research Center, KRIBB) ;
  • Yang, Sung-Hyun (Laboratory of Microbial Genomics, Systems Microbiology Research Center, KRIBB) ;
  • Choi, Soo-Keun (Laboratory of Microbial Genomics, Systems Microbiology Research Center, KRIBB) ;
  • Kim, Ji-Hyeon (Laboratory of Microbial Genomics, Systems Microbiology Research Center, KRIBB) ;
  • Kim, Jong-Guk (Department of Microbiology, College of Nature Science, Kyungpook National University) ;
  • Park, Seung-Hwan (Laboratory of Microbial Genomics, Systems Microbiology Research Center, KRIBB)
  • 발행 : 2007.03.28

초록

한반도 중부지방인 충청남북도 7개 지역에서 재배되고 있는 벼 시료 21점을 채집하여 이들의 뿌리를 표면살균 한 후 내생균을 44주 분리하고 내생성 검정 시스템을 통해 정착력이 상대적으로 우수한 균주를 최종 16주 확보 하였다. 이들의 분리빈도는 뿌리 생체중 1g당 $10^{3-5}$ CFU로 나타났다. 흥미롭게도 이중 7주가 Eurkholderia 속 균으로 동정되어 기존의 다른 벼 내생세균 연구 결과와는 다른 특징을 보였다. 또한 GFP tagging 방법을 이용하여 분리균주 중 하나인 Enterobacter sp. KJ001 균주에 대해 뿌리조직 내 colonization 위치를 확인해본 결과 뿌리 조직 중 관다발 주변에 군락을 이루고 있음을 관찰하였다. Burkholderia 분리주들은 국내 재배 벼에서 높은 빈도로 분리되며 in vitro상에서 광범위한 진균성 식물병원균에 대해 우수한 길항력과 더불어 대부분 질소고정 관련 유전자인 nifH를 가지는 점으로 보아 질소고정에 의해 식물생육에 도움을 줄 수 있을 것으로 예측되며 실제로 오이 유묘의 생장을 30% 이상 촉진하는 효과를 보여 식물병 억제 및 감소와 더불어 작물의 생장 촉진 및 생산성 증대에 활용가치가 높은 내생균으로 사료된다.

The 44 endophytic bacterial strains were isolated from surface-sterilized root of rice cultivated in seven different locations of Chungcheong province, Korea. Each isolate was introduced into rice seedlings grown gnotobiotically by inoculating scissor-cut first true leaf with cell suspensions, and the colonization capacity of each isolate in root tissue was analyzed at 7 days after inoculation. Sixteen out of 44 isolates were re-isolated from root successfully with the frequency of $10^{3-5}$ CFU/g tissue. Interestingly, seven out of 16 isolates were identified as Burkholderia species. The identity between inoculated strains and re-isolates was confirmed by genomic finger-printing and 16S rDNA sequence analysis. By a confocal laser scanning microscopic observation it was revealed that KJ001 strain, one of the sixteen isolates tagged with gfp colonized in root tissue especially around xylem. Six out of seven Burkholderia strains obtained in this study showed antagonizing activities against seven different fungal pathogens, contain nifH gene, and five of them enhanced growth of cucumber over 30%. The isolates showed no hypersensitive response on tobacco leaves and no pathogenecity in rice. From these results it was found that the endophytic Burkholderia strains will be useful in agriculture to develop a biocontrol agent or a bio-fertilizer.

키워드

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