비병원성 Erwinia carotovora subsp. carotovora 9-3의 건조 및 저장의 과정에서 생존에 미치는 염 효과

High Concentration of Sodium Chloride Increases on Survival of Non-pathogenic Erwinia carotovora subsp. carotovora 9-3 during Drying and Storage

  • 박경수 (충북대학교 농업생명환경대학 식물의학과) ;
  • 김건주 (충북대학교 농업생명환경대학 식물의학과) ;
  • 신윤주 (충북대학교 농업생명환경대학 식물의학과) ;
  • 김식 (충북대학교 농업생명환경대학 식물의학과) ;
  • 차재순 (충북대학교 농업생명환경대학 식물의학과)
  • Park, Kyoung-Soo (Department of Plant Medicine, College of Agriculture, Life and Environment Science, Chungbuk National University) ;
  • Kim, Gun-Ju (Department of Plant Medicine, College of Agriculture, Life and Environment Science, Chungbuk National University) ;
  • Shin, Yun-Ju (Department of Plant Medicine, College of Agriculture, Life and Environment Science, Chungbuk National University) ;
  • Kim, Sik (Department of Plant Medicine, College of Agriculture, Life and Environment Science, Chungbuk National University) ;
  • Cha, Jae-Soon (Department of Plant Medicine, College of Agriculture, Life and Environment Science, Chungbuk National University)
  • 발행 : 2008.12.31

초록

길항력이 있는 그람음성균들의 생물적 방제제로 성공적인 개발은 균의 건조와 저장시의 낮은 생존을 때문에 건조제형으로 만드는 것은 제한되어있다. 그람음성균의 배양시 높은 염(NaCl)의 첨가는 삼투압 조절제인 osmolyte의 생성을 유도하여 건조 시 생존율을 높인다고 알려져 있다. 그람음성의 길항력을 내는 비병원성 Erwinia carotovora subsp. carotovora(Ecc) 9-3의 건조와 저장시의 염의 영향에 관해 연구하였다. Ecc 9-3의 생장률은 0.5 M의 염이 첨가되어 있는 배지에서 큰 차이가 없었지만 0.7 M의 염이 첨가된 배지에서는 급격하게 낮아진 것을 확인하였다. 건조와 저장시의 생존율은 0.5 M의 염을 넣은 배지가 염을 넣지 않은 배지보다 상온건조와 동결건조에서 각각 2배 그리고 3배의 높은 생존율을 얻었다. 건조 후 저장에서도 역시 염이 높을수록 Ecc 9-3의 생존율이 더 높았다. 그리고 상온건조와 동결건조 모두 $4^{\circ}C$에 저장하였을 때 생존율에 큰 차이가 없었다. 그러나 생존율은 $27^{\circ}C$, $37^{\circ}C$에 저장하였을 때 염의 농도가 높을수록 생존율이 증가하였다. 첨가제 실험에서는 lactose가 동결건조와 상온건조 모두에 영향을 주었고 dextrin은 상온건조에 큰 영향을 주었다.

Dry formulation is a limiting step for successful development of microbial bio-pesticides with the antagonistic Gram-negative bacteria because their survival rates are too low during drying and storage. The high concentration of sodium chloride (NaCl) in culture medium that induces osmolyte in bacterial cells is known to increase of survival rate during drying in many Gram-negative bacteria. Effect of NaCl on survival of antagonistic non-pathogenic Erwinia carotovora subsp. carotovora 9-3 (Ecc 9-3) during drying and storage was studied. Growth rate of Ecc 9-3 was not much different up to 0.5 M NaCl in NB while it was lower significantly with 0.7 M NaCl. Survival rates were twice and 3 times higher with 0.5 M NaCl than with no additional NaCl during drying at room temperature and freeze-drying, respectively. Survival rate was also higher with high NaCl in culture medium during storage after drying. It was not much different on storage at $4^{\circ}C$ both of drying at room temperature and freeze-drying. However, the survival rate was higher on storage at $27^{\circ}C$ and $37^{\circ}C$ with high NaCl concentration. Among the additives tested, lactose affects most to survival rate both of drying at room temperature and freeze-drying, and dextrin influenced significantly to survival rate of drying at room temperature.

키워드

참고문헌

  1. Bonaterra, A., J. Camps and E. Montesinos (2005) Osmotically induced trehalose and glycine betaine accumulation improves tolerance to desiccation, survival and efficacy of the postharvest biocontrol agent Pantoea agglomerans EPS125. FEMS Microbiol. Lett. 250:1-8 https://doi.org/10.1016/j.femsle.2005.06.028
  2. Bonaterra, A., J. Cabrefiga, J. Camps and E. Montesinos (2007) Increasing survival and efficacy of a bacterial biocontrol agent of fire blight of rosaceous plants by means of osmoadaption. FEMS Microbiol Ecol. 61(1):185-195 https://doi.org/10.1111/j.1574-6941.2007.00313.x
  3. Corcoran, B. M., R. P. Ross, G. F. Fitzgerald, P. Dockery and C. Stanton (2006) Enhanced survival of GroESL- overproducing Lactobacillus paracasei NFBC 338 under stressful conditions induced by drying. Appl. Environ. Microbiol. 72(7):5104-5107 https://doi.org/10.1128/AEM.02626-05
  4. Costa, E., J. Usall, N. Teixido, R. Torres and I. Vinas (2002) Effect of package and storage condition on viability and efficacy of the freeze-dried biocontrol agent Pantoea agglomerans strain CPA-2. J. Appl. Microbiol. 92:873-878 https://doi.org/10.1046/j.1365-2672.2002.01596.x
  5. Dreux, N., C. Albagnac, R. D. Sleator, C. Hill, F. Carlin, C. E. Morris and C. Nguyen-the (2007) Glycine betaine improves Listeria monocytogenes tolerance to desiccation on parsley leaves independent of the osmolyte transporters BetL, Gbu and OpuC. J Appl Microbiol. 104(4):1221-1227 https://doi.org/10.1111/j.1365-2672.2007.03623.x
  6. Eley, A., I. Geary, A. Bahador and H. Hakimi (2006) Effect of storage temperature on survival of Chlamydia trachomatis after lyophilization. J. Clincal Microbiol. 44(7):2577-2578 https://doi.org/10.1128/JCM.00968-06
  7. Emmert, E. A. B. and J. Handelsman (1999) Biocontrol of plant disease: A (gram-)positive perspective. FEMS Microbiol, lett. 171:1-9 https://doi.org/10.1111/j.1574-6968.1999.tb13405.x
  8. Schisler, D. A., P. J. Slininger, R. W. Behle, and M. A. Jackson (2004) Formulation of Bacillus spp. for biological control of plant diseases. Phytopathoiogy, 94:1267-1271 https://doi.org/10.1094/PHYTO.2004.94.11.1267
  9. Honeycutt, E. W. and D. M. Benson. (2001) Fomulation of binucleate Rhizoctonia spp. and biocontrol of Rhizoctonia solani on impatiens. Plant Dis. 85:1241-1248 https://doi.org/10.1094/PDIS.2001.85.12.1241
  10. Prasad, J., P. McJarrow and P. Gopal (2002) Heat and osmotic stress responses of probiotic Lactobacillus rhamnosus HN001 (DR20) in relation to viability after drying. Appl. Environ. Mirobiol. 69(2):917-925
  11. Munoz-Rojas, J., P. Bernal, E. Duque, P. Godoy, A. Segura and J. L. Ramos (2005) Involvement of cyclopropane fatty acid in the response of Pseudomonas putida KT2440 to freeze- drying. Appl. Environ. Microbiol. 72(1):472-477 https://doi.org/10.1128/AEM.72.1.472-477.2006
  12. Marin, K., J. Huckauf, S. Fulda, and M. Hagemann (2002) Salt-dependent expression of glucosylglycerol-phosphate synthase, involved in osmolyte synthesis in the cynobacterium Synechocystis sp. strain PCC 6803. J. Bacteriol. 184(11): 2870-2877 https://doi.org/10.1128/JB.184.11.2870-2877.2002
  13. Zimmerman Leonard (1962) Survival of Serratia marcescens after freeze-drying or aerosolization at unfavorable humidity. J. Bacteriol. 84:1297-1302
  14. Goude, R., S. Renaud, S. Bonnassie, T. Bernard and B. Carlos (2004) Glutamine, glutamate, and -glucosylglycerate are the major osmotic solutes accumulated by Erwinia chrysanthemi strain 3937. Appl. Environ. Microbiol. 70(11):6535-6541 https://doi.org/10.1128/AEM.70.11.6535-6541.2004
  15. Wu, C and T. Hsiang (1998) Pathogenicity and formulation of Typhula phacorrhiza, biocontrol agent of gray snow mold. Plant Dis. 82(9):1003-1006 https://doi.org/10.1094/PDIS.1998.82.9.1003
  16. Silva, Z., S. Alarico, A. Nobre, R. Horlacher, J. Marugg, W. Boos, A. I. Mingote, and M. S. da. Costa. (2003) Osmotic adaptation of Thermus thermophilus RQ-1: lesson from a mutant deficient in synthesis of trehalose. J. Bacteriol. 185(20):5943-5952 https://doi.org/10.1128/JB.185.20.5943-5952.2003
  17. 김두호 (2005) 친환경농업의 현재와 미래. 한국농약과학회 학술발표대회 논문집. 10-13
  18. 김호산, 노종열, 이대원, 장진희, 제연호, 우수동, 김주경, 유용만, 강석권 (1998) 새로운 Bacillus thuringiensis NT0423 균주의 제제화. Kor. J. Appl. Microbiol. Biotechnol. 26(4):358-364
  19. 백수봉, 구항모, 도은수, 천세연 (2002) 식물병의 생물적 방제. 한국미생물생명공학회 15(4): p. 51
  20. 이동희, 조좌형, 이노운 (1992) 동결건조법이 Rifamycin 발효의 starter cell에 미치는 영향. Kor. J. Appl. Microbiol. Biotechnol. 20(4):470-476
  21. 지형진 (2007) 친환경농업과 친환경농자재에 대한 이해. 한국농약과학회 학술발표대회 논문집. 15-20
  22. 하영효 (2003) 한국유기농업학회 학술발표대회 논문집. 3-6