대파 뿌리로부터 흑색썩음균핵병균에 길항하는 Serratia plymuthica AL-1의 분리 및 Chitinase의 생산

Chitinase Production and Isolation of Serratia plymuthica AL-1 Antagonistic to White Rot Fungi from Allium fistulosum Roots.

  • 주길재 (경북대학교 농화학과) ;
  • 이익희 (상주대학교 식물자원학과) ;
  • 김진호 (상주대학교 식물자원학과)
  • 발행 : 2002.06.01

초록

대파 뿌리로부터 근권미생물 146종을 분리하여 대파 흑색썩음균핵병균인 Sclerotium cepivorum에 길항하는 AL-1 균주를 최종선별하였다. 분리주 AL-1은 the procaryotes와 Bergey's mannual of systematic bacteriology의 방법과 16S rDNA의 부분염기서열을 결정하여 ribosomal database 에서 상동성 검색 등의 방법으로 Serratia plymuthica로 동정되었다. S. plymuthica AL-1은 흑색썩음균핵병균(Sclerotium cepivorum)에 대해서는 생육저지환의 크기가 15mm로 나타났으며, 고추 검은무늬병(Alternaria altrata)은 9 mm, 고추 탄저병균(Colletotrichum gleosporioids)은 13 ㎜, 도라지 줄기마름병균(Phoma sp.)은 10 ㎜, 고추 잘록병균 (Rhizoctonia solani)은 8 ㎜, 고추 흰별무의병균(Stemphylium solani)은 8 ㎜, 오이 균핵병균(Sclerotinia sclerotiorum)은 7 mm, 수박 덩굴쪼김병(Fusarium oxysporium niveum)은 7 ㎜로 길항력을 나타내었으나 참외 만고병균(Didymella bryoniae)에서는 길항력이 없었다. S. plymuthica AL-1는 1% colloidal chitin을 첨가한 TSB 배지에서 분자량 10 kDa 이상의 분획에서는 chitinase(3.2 units/ml)가 유도 생산되었고 80℃에서 30분간 열처리할 경우 chitinase의 활성은 없어 졌으나 길항력(6.4 ㎜)은 남아있었다. 또한 분자량 10kDa 이하의 분획에서는 chitinase 활성은 없으나 길항력(5.2㎜)은 나타내었고, 80℃에서 열처리하여도 길항력(5.0mm)이 남아있어 효소 이외 다른 생리활성물질이 존재함을 확인하였다.

This study was carried out to isolate antagonistic bacterium against Sclerotium cepivorum causing Allium fistulosum white rot. Total of 146 strains were isolated from A. fistulosum roots. The isolates were screened for antagonism to S. cepivorum and the isolated strain No. AL-1 was selected among these bacteria. It was identified as Serratia plymuthica based on morphological and physiological characteristics according to the Bergey's mannual of systematic bacteriology and 16S rDNA sequences methods. Serratia plymuthica AL-1 showed broad spectrum of antifungal activities against plant pathogenic fungi Alternaria altrata, Colletotrichum gleosporioids, Phoma sp., Rhizoctonia solani, Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium niveum but not inhibited Didymella bryoniae. When S. plymuthica AL-1 cultivated in the TSB medium containing 1% colloidal chitin, the high molecular fraction (>10 kDa) have chitinase activity (3.2 units/ml) and the low molecular fraction (<10 kDa) have not chitinase activity. Oppositely, after heat treatment (80℃ for 30 min) of the cultivation supernatant, the high molecular fractions have not antifungal activity but the low molecular fractions have antifungal activity.

키워드

참고문헌

  1. Plant and Soil v.109 Differential reactions of wheat and pea genotypes to root inoculation with growth affection rhizobacteria Alstrom, B.;B. Gerhardson https://doi.org/10.1007/BF02202093
  2. J. of Plant Disease and Protection v.103 Rhizobacteria of oilseed rape antagonistic to Verticillium dahliae Berg, G.
  3. Agronomy J. v.88 Interaction involving allelopathy in cropping systems Einhelling, F. https://doi.org/10.2134/agronj1996.00021962003600060007x
  4. Mol. Plant-Microbe Interact. v.9 Suppression of Septoria tritici blotch and leaf rust of wheat by recombinant cyanide-producing strains of Pseudomonas putida Flaishman, M. A.;Z. Eyal;A. Zilberstein;C. Voisard;D. Hass https://doi.org/10.1094/MPMI-9-0642
  5. Molecular Approaches in Biological Control, IOBC/wprs Bulletin v.21 Mechanisms involved in the antifungal activity of the rhizobacterium Serratia plymuthuca Frankowski, J.;G. Berg;H. Bahl;B. K. Duffy(eds.);U. Rosenberger(eds.);G. Defago(eds.)
  6. Soil Biol. Biochem. v.25 Biological control of soilborne plant pathogens by a β-1,3-glucanase-producing Pseudomonas cepacia Fridlender, M.;J. Inbar;I. Chet https://doi.org/10.1016/0038-0717(93)90217-Y
  7. J. Gen. Microbiol. v.98 Taxonomy of the genus Serratia Grimmont, P. A. D.;F. Grimmount;H. L. C. Dulong De Ronay;P. H. A. Sneath https://doi.org/10.1099/00221287-98-1-39
  8. Bergey's manual of determinative bacteriology, 9th. Holt, J. G.;N. R. Krieg;P. H. A. Sneath;J. T. Staley;S. T. William
  9. Biological Control of Plant Diseases Progress and Challenges for the Future Nato ASI Series a v.230 Rhizosphere populations dynamics and internal colonization of cucumber by plant growth promoting rhizobacteria which induce systemic resistance to Collectotrichum orbiculare Kloepper, J. W.;G. Wie;S. Tzun;E. C. Tjamos(eds.);G. C. Papavizas(eds.);J. Cook(eds.)
  10. Appl. Environ. Microbiol. v.66 16S rRNA gene-based detection of tetrachoroethene-dechlorinating desulfuromonase and dehaloccoides species Loffler, F. E.;Q. Sun;J. Li;J. Tiedje https://doi.org/10.1128/AEM.66.4.1369-1374.2000
  11. Int. J. Food Microbiol. v.28 Incidence of histamine-forming bacteria and histamine content in scombroid fish species from retail markets in the barcelona area Lopez-Sabater, E. I.;J. J. Rodriguez-Jerez;M. Hemandez-Herrero;M. T. Moria-Ventura https://doi.org/10.1016/0168-1605(94)00007-7
  12. Eur. J. Plant Pathol. v.102 Evaluation of plant growth-promoting rhizobacteria for biological control of Pythium root of cucumbers grown in rockwool and effects on yield McCullagh, M.;R. Utkehde;J. G. Menzies;Z. K. Punja;T. C. Paulitz https://doi.org/10.1007/BF01877149
  13. Plant and Soil v.173 Survey of indigenous bacterial endophytes from cotton and sweet corn McInroy, J. A.;J. W. Kloepper https://doi.org/10.1007/BF00011472
  14. CLAO J. v.22 Microbial contamination of contact lens among medical students Midelfart, J.;A. Midelfart;L. Bevanger
  15. Anal. Chem. v.31 Use of dinitrosalicylic acid reagent for determination of reducing suger Midelfart, G. L. https://doi.org/10.1021/ac60147a030
  16. Plant Pathol. v.40 Assessment of in vivo screening systems for potential biocontrol agents of Gaeumannomyces graminis Renwick, A.;R. Campbell;S. Coe https://doi.org/10.1111/j.1365-3059.1991.tb02415.x
  17. Phytopathology v.72 Effect of Pseudomonas putida and a synthetic iron chelator on induction of soil suppressiveness to Fusarium wilt pathogens Scher, F. M.;R. Baker https://doi.org/10.1094/Phyto-72-1567
  18. Appl. Environ. Microbiol. v.58 Isolation of 2,4-diacetylphloroglucinol from a fluorescent pseudomonad and investigation of physiological paramenters influencing its production Shanahan, P.;D. J. O'Sullivan;P. Simpson;J. D. Glennon;F. O'Gara
  19. Crop Science v.36 Allelopathic effects on germination and seedling growth of clovers by endophyte-free and infected tall fescue Springer, T. L.
  20. Plant Pathology v.43 Biocontrol of post-harvest fungal dieseases on Dutch white cabbage by Pseudomonas and Serratia antagonists in storage trials Stanley, R.;M. Brown;N. Pool;D. Rodgerson;C. Sigee;C. Knight;H. Ivin;A. S. Epton;C. Leifert https://doi.org/10.1111/j.1365-3059.1994.tb01597.x
  21. A handbook and identification of bacteria Starr, M.;H. Stolp;H. Truper;H. G. Schlegel
  22. Soil Biol. & Biochem. v.33 Suppression of Sclerotinia sclerotiorum apothecial formation by the soil bacterium Serratia plymuthica:identification of a chlorinated macrolide as one of the causal agents Thaning, C.;C. J. Welch;J. J. Borowicz;R. Hedman;B. Gerhardson https://doi.org/10.1016/S0038-0717(01)00109-2
  23. FEBS Letters v.441 Proposition for the biochemical mechanism occurring in the surose isomerase active site Veronese, T.;P. Perlot https://doi.org/10.1016/S0014-5793(98)01582-8
  24. Enzyme Microb. Technol. v.24 Mechanism of sucrose conversion by the sucrose isomerase of Serratia plymuthical ATCC15928 Veronese, T.;P. Perlot https://doi.org/10.1016/S0141-0229(98)00115-X
  25. Agronomy J. v.88 Utilization of allelopathy for weed management in agroecosystems Weston L. A. https://doi.org/10.2134/agronj1996.00021962003600060004x
  26. 한국농화학회지 v.34 식물 병원성 사상균에 길항력을 갖는 Serratia marcescens CK-3의 분리 및 효소적 성질 김영일;이영환;김광식;박화성;전우복;이재화;김종현
  27. 뿌리의 활력과 근권미생물 小林達治
  28. 식물병리학 이두형;백수봉
  29. 한국농화학회지 v.42 Chitinase를 생산하는 길항미생물 Serratia sp. 3095의 선발과 Fusarium 속에 대한 항진균성 이은탁;김상달
  30. 한국생물공학회지 v.11 Chitinase를 생성하는 Serraia sp. JM의 분리 및 특성 차지명;진상기;고한철;이인화
  31. 한국원예학회지 v.40 대파의 뿌리 분비물이 국화의 생육에 미치는 영향 최상태;안형근;강석우;박인환;정우윤;장영득