Diversity of Acid-Tolerant Epiphytic Bacterial Communities on Plant Leaves in the Industrial Area and the Natural Forest Area Based on 16S rDNA

16S rDNA 염기서열에 의한 청정지역 및 공단지역 내 식물잎권의 내산성세균 군집의 다양성

  • 정필문 (대전대학교 생명과학부) ;
  • 신광수 (대전대학교 생명과학부, 대전대학교 동서생명과학연구원) ;
  • 임종순 (대전대학교 동서생명과학연구원, 대전대학교 한의학과) ;
  • 이인수 (한남대학교 미생물학과) ;
  • 박성주 (대전대학교 생명과학부, 대전대학교 동서생명과학연구원)
  • Published : 2001.12.01

Abstract

The diversity of acid-tolerant epiphytic bacterial communities on deciduous oak tree (Quercus dentate Thunb.) leaves was examined both in the natural forest area with a clean air and in the industrial estate to assess effects of acidic depositions to the phyllosphere using 16S rDNA sequence data. A total of 444 acid-tolerant epiphytic bacterial clones were obtained, resulting in 17 phylotypes by performing a analysis of restriction fragment length polymorphism (RFLP) for PCR-amplified 16S rDNA products. A very low diversity of dominating acid tolerant bacterial communities in both areas was found, just 2 subphyla groups, $\gamma$-Proteobacteria and low-G+C gram-positive bacteria. As tree leaves grow older, diversities of acid-tolerant bacteria on them significantly increased. The community structure of acid-tolerant epiphytic bacteria consisted of Pseudomonas and Enterobacteriaceae groups in the $\gamma$-Proteobacteria subphylum, and Streptococcaceae and Staphylococcus groups in the low-G+C gram-positive bacteria subphylum. The direct influence of acidic depositions on bacterial phylogenetic composition could not be detected especially when higher taxonomic levels such as subphylum, but at narrower or finer levels it could be observed by a detection of Xanthomonadales group belonged to the $\gamma$-Proteobacteria only in the industrial area and of Acetobacteraceae group belonged to the $\alpha$-Proteobacteria. There remains that these specific acid-tolerant epiphytic bacterial groups could be used as indicators for assessing effects of acidic depositions on the phyllosphere.

깨끗한 대기가 유지되는 청정지역 및 산성강하물의 영향을 받는 공단지역에서 자라는 떡갈나무(Quercus dentate Thunb.) 잎표면에서 분리한 내산성세균 배양으로부터 16S rDNA를 추출하여 모두 444개의 clone을 얻었으며, 이를 대상으로 Restriction fragment length polymorphism (RFLP)을 실시한 결과 총 17종류의 계통형 (phylotype)이 나타났다. 두 지역에서 나타난 대표적인 내산성 잎권세균 군집은 매우 단순하여 ${\gamma}$-Proteobacteria와 low-G+C gram-positive bacteria의 2개 group이었다. 식물 잎의 나이가 들수록 내산성 잎권세균 계통형의 다양성은 현저하게 증가하였다. 내산성 잎권세균 군집 구조는 $\gamma$-Proteobacteria에 속하는 Pseudomonas group과 Enterobacteriaceae, 그리고 low-G+C gram-positive bacteria 즉 Bacillus/Clostridium group에 속하는 Streptococcaceae와 Staphylococcus group이 우점하였다. 산성강하물에 따른 내산성 잎권세균 군집 구조의 변화는 상위 계통분류군(subphylum 수준)에서는 뚜렷이 볼 수 없었지만 보다 하위분류군에서 볼 때 $\gamma$-Proteobacteria의 Xanthomonadales group은 공단지역에서만, 그리고 $\alpha$-Proteobacteria의 Acetobacteraceae는 청정지역에서만 각각 검출되었으므로 이들 세균집단을 산성강서만, 그리고 $\alpha$-Proteobacteria의 Acetobacteraceae는 청정지역에서만 각각 검출되었으므로 이들 세균집단을 산성강 하물에 대한 지표세균으로서 사용할 수 있는 가능성을 남겨 놓았다.

Keywords

References

  1. Kor. J. Microbiol. v.33 Distribution of epiphytic bacteria and acid-tolerant bacteria on the phyllosphere in the industrial and clean areas Ahn, J.-H.;S.-J. Bang;N.-J. Han;W.-Y. Song;S. Hwang;I.-S. Lee;S.J. Park
  2. Nucleic Acids Res. v.25 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs Altschul, S.F;TL. Madden;A.A. Schoffer;J. Zhang;Z. Zhang;W. Miller;D.J. Lipman
  3. Aerial plant surface microbiology Phyllosphere ecology Andrews, J.H.;C.E. Morris(ed.);P.C. Nicot(ed.);C. Nguyen-The(ed.)
  4. Annu. Rev. Phytopathol. v.33 The secret life of foliar bacterial pathogens on leaves Beattie, G.A.;S.E. Lindow
  5. Bergey's manual of determinative bacteriology(9th ed.) Williams & Wilkins
  6. Appl. Environ. Microbiol. v.66 Increase in bacterial community diversity in subsurface aquifers receiving livestock wastewater input Cho, J.-C.;S.-J. Kim
  7. Methods in microbiology v.22 Methods for enumerating microorganisms and determining biomass in natural environments Hebert, R.A.;R. Grigorova(ed.);J. R. Norris(ed.)
  8. FEMS Microbiol. Ecol. v.18 A review of issues related to the quantification of bacteria from the phyllosphere Jacques, M.-A.;C.E. Morris
  9. Appl. Environ. Microbiol. v.61 Population sizes, immigration, and growth of epiphytic bacteria on leaves of different ages and positions of field-grown endive (Cichorium endivia var. latifolia) Jacques, M.-A.;L.L. Kinkel;C.E. Morris
  10. Mammalian protein metabolism Evolution of protein molecules Jukes, T.H.;C.R. Cantor;H.N. Munro(ed.)
  11. Nucleic acid techniques in bacterial systematics 16S/23S rRNA sequencing Lane, D.J.;E. Stackebrandt(ed.);M. Goodfellow(ed.)
  12. Appl. Environ. Microbiol. v.62 Nonradioactive method to study genetic profiles of natural bacterial communities by PCR single-strand-conformation polymorphism Lee, D.-H.;Y.-G. Zo;S.J. Kim
  13. Appl. Environ. Microbiol. v.44 Plants as sources of airborne bacteria, including ice nucleation-active bacteria Lindemann, J.;H.A. Constantinidou;W.R. Barchett;C.D. Upper
  14. Appl. Environ. Microbiol. v.56 Microflora partially processed lettuce Magnuson, J.A.;A.D. King, Jr.;T. Torok
  15. Molecular ecology of aquatic communities Diversity of bacterial communities in Adirondack lakes: do species assemblages reflect lake water chemistry? Methe, B.A.;J.P. Zehr;J. P. Zehr(ed.);M. A. Voytek(ed.)
  16. Mol. Ecol. v.6 Microbial aggregates in leaf surfaces: characterization and implications for the ecology of epiphytic bacteria Morris, C.E.;M.-A. Jacques;P.C. Nicot
  17. Eur. J. Soil. Biol. v.29 Succession of bacteria attached to evergreen oak leaf surfaces Perissol, C.;M. Roux;J.L. Petit
  18. Mol. Biol. Evol. v.4 The neighbor-joining nethod: a new method for reconstruction phylogenetic trees Saitou, N.;M. Nei.
  19. The prokaryotes v.1 Chapter 62: The genus Xanthomonas Starr, M.P.;M.P. Starr(ed.);H. Stolp(ed.);H.G. Truper(ed.);A. Balows(ed.);H.G. Schlegel(ed.)
  20. The prokaryotes v.Ⅰ Chapter 64: The genera Gluconobacter and Acetobacter Swings, J;J.D. Ley;M.P. Starr(ed.);H. Stolp(ed.);H.G. Truper(ed.);A. Balows(ed.);H.G. Schlegel(ed.)
  21. Mol. Ecol. v.6 Phenotypic and genotypic diversity of fluorescent pseudomonads isolated from the phyllosphere of field-grown sugar beet Thompson, I.P.;M.J. Bailey;P.B. Rainey
  22. Nucleic Acids Res. v.22 CLUSTALW; improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice Thompson, J.D.;D.G. Higgins;T.J. Gibson