DOI QR코드

DOI QR Code

Anemarrhena asphodeloides Extract Inhibits the Mycelial Growth of Magnaporthe oryzae and Controls the Rice Blast Disease

  • Joo, Myoung Ho (Department of Plant Biotechnology, Chonnam National University) ;
  • Yeo, Yu Mi (Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration) ;
  • Choi, Pil Son (Department of Oriental Pharmaceutical Development, The Nambu University) ;
  • Lee, Jae Hyeok (Department of Oriental Pharmaceutical Development, The Nambu University) ;
  • Yang, Kwang-Yeol (Department of Plant Biotechnology, Chonnam National University) ;
  • Lee, Young Jin (Department of Oriental Pharmaceutical Development, The Nambu University)
  • 투고 : 2018.11.26
  • 심사 : 2018.12.19
  • 발행 : 2018.12.31

초록

Previously, we have reported a plant extract isolated from Lysimachia foenum gracum Herba as a new environment friendly biopesticide that has the mycelial growth inhibition effect on Magnaporthe oryzae, the pathogenic fungus of the rice blast disease. For the finding of additional biopesticide candidate, we tested the mycelial growth inhibitory effects about 700 species of plant extracts on PDA media. Among them, the extract of Anemarrhena asphodeloides showed prominent inhibitory effect of which $IC_{50}$ was $139.7{\mu}g/ml$. Mycelial radii of M. oryzae were measured on PDA medium containing the four organic solvent fractions isolated from total extract from A. asphodeloides. Ethyl acetate fraction showed the impressive inhibitory effect of $IC_{50}$, $54.12{\mu}g/ml$. In the subsequent rice field test for the total extract of A. asphodeloides, we obtained encouraging 62.0% control rate of rice blast disease without any phytotoxicity. It is almost equivalent to that of chemical pesticides implying the applicability of the extract as a new biopesticide. In further study, the analysis of active ingredients of the extract would be necessary for the development of a new biopesticide and for the verification of cellular mechanism by which the mycelial growth of M. oryzae inhibited.

키워드

JOSMBA_2018_v31n6_695_f0001.png 이미지

Fig. 1. Massive screening for the mycelial growth inhibitory effects of 700 kinds of plant extracts on M. oryzae. Plant extracts provided by plant extract bank (Korea) were tested on PDA solid media in 24 well plates by empirical concentration of 500 ㎍/㎖ of each plant extracts.

JOSMBA_2018_v31n6_695_f0002.png 이미지

Fig. 2. Mycelial growth inhibitory effect of extract of A. asphodeloides on M. oryzae in dose dependent treatment. (A) None treatment. Extract was treated by the concentrations of 10, 50, 100, 150, 200, 300, 400, 500, 600, and 700 ㎍/㎖, respectively. Mycelia of M. oryzae cut by cork borer were incubated at 28℃ on the PDA solid media for 5 days. (B) The diameters or radii were determined by mean value of individual measured values. Those experiments were performed in triplicate (p<0.05, Anova test).

JOSMBA_2018_v31n6_695_f0003.png 이미지

Fig. 3. Fractionation of methanol total extract of A. Asphodeloides extract. Each fraction was prepared by orderly treatment of n-Hexan, chloroform, ethyl acetate and n-butanol and were vaporized for the treatment of mycelial growth inhibitory test.

JOSMBA_2018_v31n6_695_f0004.png 이미지

Fig. 4. Mycelial growth inhibitory effect of fraction. (A) Hexane, Chloroform, Ethyl acetate, and n-butanol fraction were tested by the concentration of 200 ㎍/㎖ of each fraction. (B) Mycelial radii of M. oryzae were measured on the PDA solid media for 5 days after the treatment of ethyl acetate fraction. Those experiments were performed in triplicate (p<0.05, Anova test).

JOSMBA_2018_v31n6_695_f0005.png 이미지

Fig. 5. View of test port of rice blast disease field test of rice plant, Chucheng. (A) whole view of testport of none treatment, (B) Symptoms of rice blast disease in none treatment test port by M. oryzae, (C) Whole view of treatment testport, (D) Test port of A. asphodeloides extract treatment. Those experiments were performed in triplicate.

Table 1. Effect of A. asphodeloides extract on control rate of rice in field test

JOSMBA_2018_v31n6_695_t0001.png 이미지

Table 2. Effect of A. asphodeloides extract on phytotoxicity of rice in field test

JOSMBA_2018_v31n6_695_t0002.png 이미지

참고문헌

  1. Chen, S., L. Wang, Z. Que and R. Pan. 2005. Genetic and physical mapping of Pi37(t), a new gene conferring resistance to rice blast in the famous cultivar St. No. 1. Theor. Appl. Genet. 111(8):1563-1570. https://doi.org/10.1007/s00122-005-0086-0
  2. Couch, B.C., I. Fudal, M.H. Lebrun, D. Tharreau, B. Valent, P. van Kim, J.L. Notteghem and L.M. Kohn. 2005. Origins of host-specific populations of the blast pathogen Magnaporthe oryzae in crop domestication with subsequent expansion of pandemic clones on rice and weeds of rice. Genetics 170(2):613-630. https://doi.org/10.1534/genetics.105.041780
  3. Cho, J.S., L.W. Chi, B.K. Jang, H.S. Jeong and C.H. Lee. 2018 Anti-microbial activities of ten Lauraceae species against Propionibacterium acnes. Korean J. Plant Res. 31(5):423-432. https://doi.org/10.7732/KJPR.2018.31.5.423
  4. Davidson, A.L., E. Dassa, C. Orelle and J. Chen. 2008. Structure, function, and evolution of bacterial ATP-binding cassette systems. Microbiol. Mol. Biol. Rev. 72(2):317-364. https://doi.org/10.1128/MMBR.00031-07
  5. Dean, M., A. Rzhetsky and R. Allikmets. 2001. The human ATP-binding cassette (ABC) transporter superfamily. Genome Res. 11(7):1156-1166. https://doi.org/10.1101/gr.GR-1649R
  6. Dean, R.A., N.J. Talbot, D.J. Ebbole, M.L. Farman, T.K. Mitchell, M.J. Orbach, M. Thon, R. Kulkarni, J.R. Xu, H. Pan, N.D. Read, Y.H. Lee, I. Carbone, D. Brown, Y.Y. Oh, N. Donofrio, J.S. Jeong, D.M. Soanes, S. Djonovic, E. Kolomiets, C. Rehmeyer, W.Li,M. Harding, S. Kim, M.H. Lebrun, H. Bohnert, S. Coughlan, J. Butler, S. Calvo, L.J. Ma, R. Nicol, S. Purcell, C. Nusbaum, J.E. Galagan and B.W. Birren. 2005. The genome sequence of the rice blast fungus Magnaporthe grisea. Nature 434(7036):980-986. https://doi.org/10.1038/nature03449
  7. Di Pietro, A., G. Dayan, G. Conseil, E. Steinfels, T. Krell, D. Trompier, H. Baubichon-Cortay and J. Jault. 1999. P-glycoprotein-mediated resistance to chemotherapy in cancer cells: using recombinant cytosolic domains to establish structure-function relationships. Braz. J. Med. Biol. Res. 32(8):925-939. https://doi.org/10.1590/S0100-879X1999000800001
  8. Dixon, R.A., M.J. Harrison and C.J. Lamb. 1994. Early events in the activation of plant defense responses. Annu. Rev. Phytopathol. 32(1):479-501. https://doi.org/10.1146/annurev.py.32.090194.002403
  9. Dixon, R.A. and C.J. Lamb. 1990. Molecular communication in interactions between plants and microbial pathogens. Annu. Rev. Phytopathol. 41(1):339-367.
  10. Froyd, J.D., C.J. Paget, L.R. Guse, B.A. Dreikorn and J.L. Pafford. 1976. Tricyclazole: a new systemic fungicide for control of Pyricularia oryzae on rice. Phytopathology 66(1):135-131. https://doi.org/10.1094/Phyto-66-1135
  11. Gulshan, K. and W.S. Moye-Rowley. 2007. Multidrug resistance in fungi. Eukaryot. Cell 6(11):1933-1942. https://doi.org/10.1128/EC.00254-07
  12. Iida, Y., K.B. Oh, M. Saito, H. Matsuoka, H. Kurata, M. Natsume and H. Abe. 1999. Detection of antifungal activity in Anemarrhena asphodeloides by sensitive BCT method and isolation of its active compound. J. Agric. Food Chem. 47(2):584-587. https://doi.org/10.1021/jf980707t
  13. Jones, P.M. and A.M. George. 2004. The ABC transporter structure and mechanism: perspectives on recent research. Cell Mol. Life Sci. 61(6):682-699. https://doi.org/10.1007/s00018-003-3336-9
  14. Jang B.K., L.W. Chi, J.S. Cho and C.H. Lee. 2018. Antimicrobial activity screening of sixty-four evergreen woody species according to extraction conditions against Trichophyton mentagrophytes. Korean J. Plant Res. 31(4):330-341. https://doi.org/10.7732/KJPR.2018.31.4.330
  15. Koh, Y.J., B.K. Hwang and H.S. Chung. 1986. Screening of rice cultivars for adult-plant resistance to Pyricularia oryzae. Korean J. Plant Pathol. 2(2):69-81.
  16. Lee, Y.J. 2016. Lysimachia foenum-graecum Herba extract, a novel biopesticide, inhibits ABC transporter genes and mycelial growth of Magnaporthe oryzae. Korean J. Plant Pathol. 32(1):8-15. https://doi.org/10.5423/PPJ.OA.08.2015.0157
  17. Lee, Y.J., K. Yamamoto, H. Hamamoto, R. Nakaune and T. Hibi. 2005. A novel ABC transporter gene ABC2 involved in multidrug susceptibility but not pathogenicity in rice blast fungus, Magnaporthe grisea. Pestic. Biochem. Phys. 81(1):13-23. https://doi.org/10.1016/j.pestbp.2004.07.007
  18. Li, G. and Y.Q. Wang. 2006. Progress in the research on the ABCA gene family of vertebrates. Yi Chuan. 28(8):1015-1022.
  19. Magaldi, S., S. Mata-Essayag, C. Hartung de Capriles, C. Perez, M.T. Colella, C. Olaizola and Y. Ontiveros. 2004. Well diffusion for antifungal susceptibility testing. Int. J. Infect. Dis. 8(1):39-45. https://doi.org/10.1016/j.ijid.2003.03.002
  20. Park, H.J., J.Y. Lee, S.S. Moon and B.K. Hwang. 2003. Isolation and anti-oomycete activity of nyasol from Anemarrhena asphodeloides rhizomes. Phytochemistry 64(5):997-1001. https://doi.org/10.1016/S0031-9422(03)00462-X
  21. Ribot, C., J. Hirsch, S. Balzergue, D. Tharreau, J.L. Notteghem, M.H. Lebrun and J.B. Morel. 2008. Susceptibility of rice to the blast fungus, Magnaporthe grisea. Plant Physiol. 165(1):114-124. https://doi.org/10.1016/j.jplph.2007.06.013
  22. Skamnioti, P. and S.J. Gurr. 2009. Against the grain: safeguarding rice from rice blast disease. Trends Biotechnol. 27(3):141-150. https://doi.org/10.1016/j.tibtech.2008.12.002
  23. Valgas, C., S.M.d. Souza, E.F. Smania and A. Smania Jr. 2007. Screening methods to determine antibacterial activity of natural products. Braz. J. Microbiol. 38(2):369-380. https://doi.org/10.1590/S1517-83822007000200034
  24. Zolnerciks, J.K., E.J. Andress, M. Nicolaou and K.J. Linton. 2011. Structure of ABC transporters. Essays Biochem. 50(1):43-61. https://doi.org/10.1042/bse0500043