Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Structural Identification of Antibiotics from Pseudomonas sp. RRj 228, a Antifungal Activity of Collectotrichum acutatum Causing Anthracnose on Pepper

Pseudomonas sp. RRj 228이 분비하는 항균물질의 동정과 고추탄저균 C. acutatum에 대한 항균활성

  • Jeon, Sang-Yoon (Department of Life Science & Environmental Biochemistry, Pusan National University) ;
  • Kim, Yong-Gyun (Department of Life Science & Environmental Biochemistry, Pusan National University) ;
  • Lee, Sang-Mong (Department of Life Science & Environmental Biochemistry, Pusan National University) ;
  • Son, Hong-Joo (Department of Life Science & Environmental Biochemistry, Pusan National University) ;
  • Park, Hyean-Cheal (Department of Life Science & Environmental Biochemistry, Pusan National University) ;
  • Kim, Sun-Tae (Department of Plant Bioscience, Pusan National University) ;
  • Park, Ki-Do (National Institute of Crop Science, RDA.) ;
  • Kang, Ui-Gum (National Institute of Crop Science, RDA.) ;
  • Kim, Keun-Ki (Department of Life Science & Environmental Biochemistry, Pusan National University)
  • Received : 2010.07.07
  • Accepted : 2010.08.18
  • Published : 2010.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

Microorganisms near the plant rhizosphere usually inhabit the surface or the inside of the plant roots and have a direct effect on plant growth by secreting plant growth promoters or antagonistic materials which protect the root zone system from various pathogens. This study was carried out to identify and isolate the antagonistic materials after isolation of microorganisms showing high antagonistic activities, in hopes of contributing to the development of sustainable agriculture and the preservation of agricultural environments. A number of antagonistic bacteria were isolated from paddy soil. Among isolates, RRj 228 showed plant growth promotion and antagonistic activity. RRj 228 was identified as Pseudomonas sp. according to the results of physiological properties and genetic methods. On the basis of the results of anti-fungal spectrum against several pathogens by RRj 228, the antagonistic effect of the isolate against Botrytis cinerea, Pythium ultimum, Phytopthola capsici, and Rhizoctonia solani, especially against red-pepper anthracnose caused by Colletotrichum acutatum, was remarkable. The experiment evaluating the biological control effect by RRj 228 revealed that the $ED_{50}$ value by the RRj 228 culture against C. acutatum, R. solani and P. ultimum were 0.14 mg/ml, 0.16 mg/ml and 0.29 mg/ml, respectively. An antagonistic substance was isolated and purified by several chromatographies from the RRj 228 culture. The $^1H$ and $^{13}C$ assignment of the antagonistic substance was achieved from two-dimensional $^1H-^1H$ COSY, HMQC, and HMBC. Finally, the antagonistic substance was identified as Phenazine-1-carboxylic acid ($C_{13}H_8N_2O_2$, M.W.=224).

식물 근권미생물은 식물의 뿌리 표면과 내부에 정착하면서 식물생장 촉진물질을 분비하여 식물생육에 직접적인 영향을 주거나, 항균물질을 분비하여 병원균으로부터 식물 근권환경을 보호하여 식물의 생육을 촉진하게 된다. 이러한 활성을 갖고 있는 미생물을 분리하여 동정하며, 항균물질을 분리, 동정하여 지속농업의 발전과 농업환경의 보전을 위하여 연구를 실시하였다. 논토양으로부터 식물생육촉진활성과 항균활성을 갖는 RRj 228균주를 분리하여, 생리 생화학적방법과 유전학적 방법으로 동정한 결과, Pseudomonas sp.로 동정되었다. RRj 228균주는 B. cinerea, P. ultimum, P. capsici와 R. solani에도 높은 항균활성을 보였으며, 특히 고추탄저병원균인 C. acutatum에 강력한 항균활성을 나타냈다. RRj 228균의 배양여액을 C. acutatum, R. solani 및 P. ultimum에 대한 $ED_{50}$값을 측정한 결과 0.14, 0.16, 0.29 mg/ml로 나타났다. 항균활성물질은 RRj 228 배양여액으로부터 각종 크로마토그라피법으로 순수분리하고, NMR과 GC/MS 등의 기기분석을 실시하여 구조를 Phenazine-1-carboxylic acid (분자량 224)로 동정하였다.

Keywords

References

  1. Ahn, I. P., K. H. Uhm, S. Kim, and Y. H. Lee. 2003. Signaling pathways involved in preinfection development of Colletotrichum gloeosporioides, C. coccodes, and C. dematium pathogenic on red pepper. Physiol. Mol. Plant Pathol. 63, 281-289. https://doi.org/10.1016/j.pmpp.2004.01.001
  2. Ahn, I. P., S. Kim, W. B. Choi, and Y. H. Lee. 2003. Calcium restores prepenetration morphogenesis abolished by polyamines in Colletotrichum gloeosporioides infecting red pepper. FEMS Microbiol. Lett. 227, 237-241. https://doi.org/10.1016/S0378-1097(03)00686-4
  3. Ahn, S. M., D. S. Lee, M. S. Kim, S. J. Choi, C. S. Choi, J. B. Lee, H. S. Jang, and H. Y. Sohn. 2009. Bioactivity of the extract of Coptis chinensis: In-vitro antifungal activity against Phytophthora capsici and growth-promotion effect in red-pepper. Kor. J. Microbiol. Biotechnol. 37, 280-286.
  4. Bardas, G. A., A. L. Lagopodi, K. Kadoglidou, and T. K. Katina. 2009. Biological control of three Colletotrichum lindemuthianum races using Pseudomonas chlororaphis PCL1391 and Pseudomonas fluorescens WCS365. Biological Control 49, 139-145. https://doi.org/10.1016/j.biocontrol.2009.01.012
  5. Chanchaichaovivat, A., P. Ruenwongsa, and B. Panijpan. 2007. Screening and identification of yeast strains from fruits and vegetables: Potential for biological control of postharvest chilli anthracnose (Colletotrichum capsici). Biological Control 42, 326-335. https://doi.org/10.1016/j.biocontrol.2007.05.016
  6. Chung, J. W., C. Y. Lee, S. C. Yun, and K. Y. Cho. 2008. Screening of myxobacteria inhibiting the growth of Collectotrichum acutatum causing anthracnose on pepper. Kor. J. Microbiol. Biotechnol. 36, 21-27.
  7. Han, K. S, J. H. Park, Y. K. Han, and J. H. Hwang. 2009. Pathogenicity and occurrence of pepper seedling anthracnose caused by Colletotrichum acutatum. Res. Plant Dis. 15, 88-93. https://doi.org/10.5423/RPD.2009.15.2.088
  8. Kang, K., K. Lee, A. Ishihara, S. Park, Y. S. Kim, and K. Back. 2010. Induced synthesis of caffeoylserotonin in pepper fruits upon infection by the anthracnose fungus, Colletotrichum gloeosporioides. Scientia Horticulturae 124, 290-293. https://doi.org/10.1016/j.scienta.2009.12.036
  9. Kim, J. T., S. K. Park, W. B. Choi, Y. H. Lee, and H. T. Kim. 2003. Identification of Colletotrichum spp. associated with pepper anthracnose in Korea. (Abst.) Plant Pathol. J. 19, 331.
  10. Kim, K. K., K. H. Park, S. S. Moon, and K. Y. Kang. 1997. Isolation and structure identification of antifungal substance from Aspergillus terreus. Agri. Chem. Biotechnol. 40, 593-596.
  11. Kim, Y. S., H. H. Lee, M. K. Ko, C. E. Song, C. Y. Bae, Y. H. Lee, and B. J. Oh. 2001. Inhibition of fungal appressorium formation by pepper (Capsicum annuum) esterase. Mol. Plant-Microbe Interact. 14, 80-85. https://doi.org/10.1094/MPMI.2001.14.1.80
  12. Kim, K. K., Y. K. Kim, H. J. Son, Y. W. Choi, and K. Y. Kang. 2005. Biological control of perilla culture by Burkhoderia sp. AK-17. J. Korean Soc. Appl. Biol. Chem. 48, 34-39.
  13. Lee, J. B., J. H. Shin1, J. O. Jang, K. S. Shin, C. S. Choi, K. W. Kim, M. S. Jo, C. P. Jeon, Y. H. Kim, and G. S. Kwon. 2008. Antifungal activity of Bacillus sp. AM-651 against Phytophthora capsici. Kor. J. Microbiol. Biotechnol. 36, 227-232.
  14. Lee, K. H. 2008. Improvement of chemical control method of anthracnose, Rural Development Administration, Region Specialization Project Report.
  15. Liu, W. H., M. G. Li, Y. Q. Li, J. Y. Zhao, Z. G. Ding, P. W. Yang, X. L. Cui, and M. L. Wen. 2008. Cytotoxic metabolites of Streptimonospora salina. Chem. Nat. Comp. 44, 503-505. https://doi.org/10.1007/s10600-008-9102-3
  16. Ozgonen, H. and A. Erkilic. 2007. Growth enhancement and phytophthora blight (Phytophthora capsici Leonian) control by arbuscular mycorrhizal fungal inoculation in pepper. Crop. Protection 26, 1682-1688. https://doi.org/10.1016/j.cropro.2007.02.010
  17. Paik, S. B. and D. W. Kim. 1995. Screening for phyllospheral antagonistic microorganisms for control of red-pepper anthracnose (Collectotrichum gloeosporioides). Kor. J. Mycol. 23, 190-195.
  18. Palleroni, N. J. 1989. Family I. Pseudomonadaceae, In BERGEY's Manual Systemic Bacteriology. 1. pp. 141-218. N. R. KRIEG, (eds.) Williams & Wilkins.
  19. Park, K. S. and C. H. Kim. 1992. Identification, distribution and etiological characteristics of anthracnose fungi of red pepper in Korea. Kor. J. Plant Pathol. 8, 61-69.
  20. Park, S. M., H. J. Jung, and T. S. Yu. 2006. Screening of an antagonistic bacterium for control of red-pepper anthracnose, Colletotrichum gloeosporioides. J. Life Science 16, 420-426. https://doi.org/10.5352/JLS.2006.16.3.420
  21. Rawlings, S. L., R. J. O'Connell, and J. R. Green. 2007. The spore coat of the bean anthracnose fungus Colletotrichum lindemuthianum is required for adhesion, appressorium development and pathogenicity. Physiol. Mol. Plant Pathol. 70, 110-119. https://doi.org/10.1016/j.pmpp.2007.07.007
  22. Sang, M. K., S. C. Chun, and K. D. Kim. 2008. Biological control of phytophthora blight of pepper by antagonistic rhizobacteria selected from a sequential screening procedure. Biological Control 46, 424-433. https://doi.org/10.1016/j.biocontrol.2008.03.017
  23. Woo, S. M. and S. D. Kim. 2008. Structural identification of siderophore (AH18) from Bacillus subtilis AH18, a biocontrol agent of phytophthora blight disease in red-pepper. Kor. J. Microbiol. Biotechnol. 36, 326-335.
  24. Yucel, S. 1995. A study on soil solarization and combined with fumigant application to control phytophthora crown blight (Phytophthora capsici Leonian) on peppers in the East Mediterranean region of Turkey. Crop Protection 14, 653-655. https://doi.org/10.1016/0261-2194(95)00057-7
  25. Zhang, S., T. L. White, M. C. Martinez, J. A. McInroy, J. W. Kloepper, and W. Klassen. 2010. Evaluation of plant growth-promoting rhizobacteria for control of phytophthora blight on squash under greenhouse conditions. Biological Control 53, 129-135. https://doi.org/10.1016/j.biocontrol.2009.10.015

Cited by

  1. Antagonistic Activities of Bacillus spp. Strains Isolated from Tidal Flat Sediment Towards Anthracnose Pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea vol.31, pp.2, 2015, https://doi.org/10.5423/PPJ.OA.03.2015.0036