part.6 - CRDC

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116 สารออกฤทธิ์ตานเชื้อราที่ทําใหเกิดโรคพืช ปที่ 40 ฉบับที่ 1 (พิเศษ) มกราคม-เมษายน 2552 ว. วิทยาศาสตรเกษตร Table 1. The antifungal activities of endophytic fungi isolated from Stemona spp. grown in modified MID for 7 and 14 days of cultivation. Isolate No. Inhibition zone (cm) Specie of Ab Penicillium sp. Fo Fs host 7 days 14 days 7 days 14 days 7 days 14 days 7 days 14 days LL-SB 49-1 S. burkillii 2.2±1.1 1.9±0.1 1.4±0.1 ND 1.3±0.1 ND 1.7±0.8 ND MP-SB 41-2 S. burkillii 1.8±0.9 ND ND ND 1.5±0.2 ND 1.7±0.8 ND LL-SB 17-1 S. burkillii 2.3±0.3 1.7±0.0 ND ND 1.4±0.2 ND 1.3±0.0 1.5±0.8 ML-SB 16-3 S. burkillii 1.9±0.1 1.6±0.1 ND ND 1.5±0.1 1.4±0.1 1.5±0.0 1.5±0.0 MP-SB 16-2 S. burkillii 2.1±1.0 1.9±0.9 1.6±0.1 ND 1.4±0.2 ND 1.4±0.0 ND ML-SB 16-1 S. burkillii 2.3±0.1 1.9±0.1 1.6±0.1 ND 1.3±0.1 ND ND ND LL-SB 30-2 S. burkillii 2.2±0.1 1.8±0.1 1.8±0.1 ND 1.5±0.1 1.4±0.1 1.6±0.2 ND SERS-P 1/1-7 S. colinsae 1.6±0.8 1.7±0.1 1.7±0.2 ND ND ND ND ND SERS-L 1/10-3 S. colinsae ND 1.7±0.1 1.5±0.1 ND ND ND ND ND SERS-L 4/7-4 S. colinsae 2.2±1.1 1.9±0.1 ND 1.5±0.0 1.5±0.1 ND ND ND RYP-SB 4-8 S. colinsae 2.1±0.1 1.8±0.2 1.5±0.1 ND 1.3±0.1 ND 1.1±0.1 ND RYR-SB 6-4 S. colinsae 2.0±0.0 1.9±0.1 1.7±0.1 ND ND ND ND ND NHL-L 6/6 S. burkillii 2.7±0.1 2.0±0.3 3.8±0.1 3. 5±0.2 7.5±1.1 5.3±0.2 4.3±0.3 2.6±0.4 Note: ND : Not Detectable Ab : Alternaria brassicola, Fs : Fusarium solani and Fo : Fusarium oxysporum Interestingly, strain NHL-L 6/6, isolated from S. burkillii, showed very high activity against all pathogens. The inhibition zone diameters of 7-day old culture liquid of NHL-L 6/6 against Ab, Penicillium sp., Fo and Fs were found to be 2.7, 3.8, 7.5 and 4.3 cm, respectively (Table 1). Thus this fungus must have a potential for agricultural and biotechnological application. There was no reports on studies of bioactive compounds in leave extracts of Stemona spp., including S. burkillii. Many biologically active compounds were found in root extracts in Stemona spp. (e.g. Pacher et al., 2002; Kostecki et al., 2004; Brem et al., 2002). In 2004, Mungkornasawakul and co-workers reported that dihydrostemofoline alkaloid from S. burkillii roots did not show significant activity on inhibiting spore germination on Cladosporium cladosporiodes. Therefore, structures of the active metabolites in NHL-L 6/6 must be elucidated to study the in vivo production of these active compounds by the fungus and to demonstrate mutalistic relationships of secondary metabolite biosynthesis in host and their symbionts. Based on the sizes of inhibition zones of the filtered culture broth, incubation period played a crucial role in secondary metabolite stability and production in these active strains because fungal inhibitory activities were significantly decreased when incubation period was increased to 14 days. This could be due to the active compounds were chemically modified by the fungi themselves or the environment change such as dissolved oxygen and pH of the culture. Physiological study of some strains such as NHL-L 6/6 for optimization of submerged culture condition should be carried to improve production of the antifungal agents in large-scale production.

ว. วิทยาศาสตรเกษตร ปที่ 40 ฉบับที่ 1 (พิเศษ) มกราคม-เมษายน 2552 สารออกฤทธิ์ตานเชื้อราที่ทําใหเกิดโรคพืช 117 Summary This work showed that endophytic fungi are a promising source of new natural bioactive agents for plant disease control. Moreover, plant in family Stemonaceae give good oppurtunities to isolate active endophytic fungi. In this work strain NHL-L 6/6 surprisingly showed very large inhibition zone against all tested pathogens when grown in modified MID medium. The role of NHL-L 6/6 within S. burkillii is still unknown. The structures of bioactive compounds synthesized by this endophyte should be characterized to study the relationship of secondary metabolite production in these two organisms. Benefits to the host plant from antagonism towards pathogenic fungi could be speculated. Investigations on the interactions of S. burkillii and its endophyte would be the next direction for future research. Literature cited Boonmakad, C., A. Supatrakul and T. Vichitsoonthonkul. 2007. Bioactive Compound Produced by Endophytic Fungai of Stemona spp. for Bacterial Plant Pathogen Control. Agricultural Sci. J. 38: 339-343. Brem, B., C. Seger, T. Pacher, O. Hofer, S. Vajrodaya and H. Greger. 2002. Feeding deterrence and contact toxicity of Stemona alkaloid- a source of potent natural insecticides. Journal of Agricultural and Food Chemistry. 50: 6383-6388. Dreyfuss, M.M. and I. Chapela. 1994. Potential of fungi in the discovery of novel, low-molecular weight pharmaceuticals. In The Discovery of Natural Products with Therapeutic Potential (ed. V. P. Gullo), 49-80. Butterworth-Heinemann: Newton, Massachusetts. Kostecki, K., D. Engelmeiera, T. Pachera, O. Hofer, S. Vajrodaya and H. Greger. 2004. Dihydro-phenanthrenes and other antifungal stilbenoids from Stemona cf. pierrei. Phytochemistry. 65: 99–106. Lingham, R.B., K.C. Silverman, G.F. Bills, C. Cascales, M. Sanchez, R.G. Jenkins, S.E. Gartner, I. Martin, M.T. Diez, F. Pelaez, S. Mochales, Y.L. Kong, R.W. Burg, M.S. Meinz, L. Huang, M. Nallin-Omstead, S.D. Mosser, M.D. Schaber, C.A. Omer, D.L., Pompiano, J.B. Gibbs and S.B. Singh. 1993. Chaetomella acutiseta produces chaetomellic acids A and B which are reversible inhibitors of farnesyl- protein transferase. Applied Microbiology and Biotechnology. 40: 370-374. Mungkornasawakul, P., G.S. Pyne, A. Jatisatienr, W. Lie, T.A. Ung, K. Issakul, A. Sawatwanich, D. Supyen and C. Jatisatienr. 2004. Phytochemical Studies on Stemona burkillii Prain: Two New Dihydrostemofoline Alkaloids. Journal of Natural Products. 67: 1740-1743. Pacher, T., C. Seger, D. Engelmeier, S. Vajrodaya, O. Hofer and H. Greger. 2002. Antifungal stilbenoids from Stemona collinsae. J. Nat. Prod. 65: 820-827. Pinkerton, F. and G. Strobel. 1976. Serinol as an activator of toxin production in attenuated cultures of Helminthosporium sacchari. Proc. Natl. Acad. Sci. USA. 73: 4007-4011. Soytong, K., V. Rakvidhvasastra and T. Sommartya. 1985. Effect of some medicinal plants on growth fungi and potential in plant disease control. Abstracts of the 11th Conference of Science and Technology Thailand. Kasetsart University. Bangkok. Thailand. October 24-26. 361 p. Zhao, W.M., G.W. Qin, Y. Ye, R.S. Xu and X.F. Le. 1995. Bibenzyl from Stemona tuberose. Phytochemistry. 38: 711-713.

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crdc

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