The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
권호 표지
DOI QR코드

DOI QR Code

Evaluation of Rhizobacterial Isolates for Their Antagonistic Effects against Various Phytopathogenic Fungi

식물 근권에서 분리한 미생물의 식물병원성 진균에 대한 길항효과 검정

  • Kim, Yun Seok (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture) ;
  • Kim, Sang woo (Division of Biological Resources Sciences, Department of Applied Plant Sciences, Kangwon National University) ;
  • Lamsal, Kabir (Division of Biological Resources Sciences, Department of Applied Plant Sciences, Kangwon National University) ;
  • Lee, Youn Su (Division of Biological Resources Sciences, Department of Applied Plant Sciences, Kangwon National University)
  • 김윤석 (농림수산식품기술기획평가원) ;
  • 김상우 (강원대학교 식물자원응용공학과) ;
  • 거비르 람살 (강원대학교 식물자원응용공학과) ;
  • 이윤수 (강원대학교 식물자원응용공학과)
  • Received : 2015.11.20
  • Accepted : 2016.03.18
  • Published : 2016.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

This study was conducted to evaluate five different strains of rhizobacterial isolates viz. PA1, PA2, PA4, PA5 and PA12 for biological control against Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor and Fusarium sp. In vitro inhibition assay was performed on three different growth mediums, potato dextrose agar (PDA), tryptic soy agar (TSA), and PDA-TSA (1:1 v/v) for the selection of potential antagonistic isolates. According to the result, isolate PA2 showed the highest inhibitory effect with 65.5% against C. coccodes on PDA and with 96.5% against S. minor on TSA. However, the same isolate showed the highest inhibition with 58.5% against C. acutatum on PDA-TSA. In addition, an in vivo experiment was performed to evaluate these bacterial isolates for biological control against fungal pathogens. Plants treated with bacteria were analyzed with phytopathogens and plants inoculated with phytopathogens were treated with isolates to determine the biological control effect against fungi. According to the result, all five isolates tested showed inhibitory effects against phytopathogens at various levels. Mode of action of these rhizobacterial isolates was evaluated with siderophore production, protease assay, chitinase assay and phosphate solubilizing assay. Bacterial isolates were identified by 16S rDNA sequencing, which showed that isolates PA1 and PA2 belong to Bacillus subtilis, whereas, PA4, PA5, and PA12 were identified as Bacilus altitudinis, Paenibacillus polymyxa and Bacillus amyloliquefaciens, respectively. Results of the current study suggest that rhizobacterial isolates can be used for the plant growth promoting rhizobacteria (PGPR) effect as well as for biological control of various phytopathogens.

본 연구는 식물 근권에서 분리한 유용미생물 PA1, PA2, PA4, PA5, PA12 의 식물 생장 촉진능력과 식물 병원성 진균인 Colletotrichum acutatum, C. coccodes, C. gloeosporioides, C. dematium, Botrytis cinerea, Rhizoctonia solani, Sclerotinia minor 그리고 Fusarium sp.에 대한 생장억제능력을 평가하는데 그 목적이 있다. In vitro 실험에서 유용미생물의 식물 병원성 진균의 생장억제 능력을 확인하기 위해 세균배지인 TSA 배지와 곰팡이 배지인 PDA배지, 그리고 TSA와 PDA배지를 각각 50%씩 혼합한 배지(v/v, 1:1)에서 대치배양을 실시하였다. 그 결과 PDA배지에서는 PA2가 C. coccodes에 대해 65.5%로 가장 높은 억제능력을 보였으며, TSA배지에서는 PA2가 S. minor에 대해 96.5%로 가장 높은 억제력을 보였다. 또한 PDA와 TSA를 혼합한 배지에서는 PA2가 C. acutatum에 대해 58.5%로 가장 높은 억제능력을 보였다. 분리한 5균주 모두에서 식물병원성 진균에 대하여 생물적 방제 효과가 있음을 확인하였다. 또한 식물생장 촉진능력을 유발하는 원인물질을 탐색하기 위해 siderophore, protease, chitinase, hydrogen cyanide (HCN) 생성 유무를 확인하였고, phosphate solubilizing 실험을 실시하였다. 본 연구에서 사용된 유용미생물 5균주를 16s rDNA sequencing 결과 PA1, PA2는 Bacillus subtilis, PA4, PA5, PA12 각각 Bacillus altitudinis, Paenibacillus polymyxa, Bacillus amyloliquefaciens로 동정되었다.

Keywords

References

  1. Liu M, Cai QX, Liu HZ, Zhang BH, Yan JP, Yuan ZM. Chitinolytic activities in Bacillus thuringiensis and their synergistic effects on larvicidal activity. J Appl Microbiol 2002;93:374-9. https://doi.org/10.1046/j.1365-2672.2002.01693.x
  2. Kai M, Effmert U, Berg G, Piechulla B. Volatiles of bacterial antagonists inhibit mycelial growth of the plant pathogen Rhizoctonia solani. Arch Microbiol 2007;187:351-60. https://doi.org/10.1007/s00203-006-0199-0
  3. Verhagen BW, Trotel-Aziz P, Couderchet M, Hofte M, Aziz A. Pseudomonas spp.-induced systemic resistance to Botrytis cinerea is associated with induction and priming of defense responses in grapevine. J Exp Bot 2010;61:249-60. https://doi.org/10.1093/jxb/erp295
  4. Niu DD, Liu HX, Jiang CH, Wang YP, Wang QY, Jin HL, Guo JH. The plant growth-promoting rhizobacterium Bacillus cereus AR156 induces systemic resistance in Arabidopsis thaliana by simultaneously activating salicylate- and jasmonate/ethylene- dependent signaling pathways. Mol Plant Microbe Interact 2011;24:533-42. https://doi.org/10.1094/MPMI-09-10-0213
  5. Roberts WK, Selitrennikoff CP. Plant and bacterial chitinases differ in antifungal activity. J Gen Microbiol 1988;134:169-76.
  6. Pikovskaya RI. Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Mikrobiologiya 1948;17:362-70.
  7. Schwyn B, Neilands JB. Universal chemical assay for the detection and determination of siderophores. Anal Biochem 1987; 160:47-56. https://doi.org/10.1016/0003-2697(87)90612-9
  8. Ahmed AS, Ezziyyani M, Perez Sanchez C, Candela ME. Effect of chitin on biological control activity of Bacillus spp. and Trichoderma harzianum against root rot disease in pepper (Capsicum annuum) plants. Eur J Plant Pathol 2003;109:633-7. https://doi.org/10.1023/A:1024734216814
  9. Rodriguez H, Fraga R. Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnol Adv 1999; 17:319-39. https://doi.org/10.1016/S0734-9750(99)00014-2
  10. Mori K, Lee HT, Rapoport D, Drexler IR, Foster K, Yang J, Schmidt-Ott KM, Chen X, Li JY, Weiss S, et al. Endocytic delivery of lipocalin-siderophore-iron complex rescues the kidney from ischemia-reperfusion injury. J Clin Invest 2005;115: 610-21. https://doi.org/10.1172/JCI23056
  11. Devi KK, Seth N, Kothamasi S, Kothamasi D. Hydrogen cyanide-producing rhizobacteria kill subterranean termite Odontotermes obesus (Rambur) by cyanide poisoning under in vitro conditions. Curr Microbiol 2007;54:74-8. https://doi.org/10.1007/s00284-006-0473-z
  12. Kremer RJ, Souissi T. Cyanide production by rhizobacteria and potential for suppression of weed seedling growth. Curr Microbiol 2001;43:182-6. https://doi.org/10.1007/s002840010284