Cultural characteristics of Cordyceps militaris strain ‘Yedang 3’ on various media and nutritional conditions
(Received September 13, 2013 / Revised September 21, 2013 / Accepted September 27, 2013)
Abstract
The effects of environmental and nutritional requirement such as temperature, pH, different media,carbon, nitrogen, and carbon-to-nitrogen ratio on the mycelial growth of Cordyceps militaris strain ‘Yedang 3’ werestudied. The optimum temperature and pH for the growth of mycelium were 20-25℃ and pH 6-7, respectively.Out of ten media tested, mushroom complete media (MCM) was the best medium for fast mycelial growth, andSabouraud’s dextrose agar yeast extract (SDAY), malt extract yeast extract agar (YMA) also were favored. Thecolor and shape of colonies varied in different media. The best carbon sources for mycelial growth were fructose,mannitol, and sucrose, whereas the best nitrogen sources were tryptone and peptone. However, mycelia grew slowlyin inorganic nitrogen compounds such as NH4Cl, (NH4)2SO4, NH4NO3, and NaNO3. The optimum C:N ratioobserved on the culture media was 30-40 range. These results provided basic information on cultural characteristicsof vegetative growth and might be useful for spawn production in Cordyceps militaris.
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Reference
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16.Liang, Z. Q. 2001. current situation and ponderation of Cordyceps Fr. research and exploitation in China. Acta Edulis Fungi 8 : 53-62.
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29.Shrestha, B., Lee, W. H., Han, S. K, and Sung, J. M. 2006. Observations on some of the mycelial growth and pigmentation characteristics of Cordyceps militaris isolates. Mycobiol. 34 : 83-91.
30.Stensrud, Ø, Hywel-Jones, N. L., and Schumacher, T. 2005. Towards a phylogenetic classification of Cordyceps: ITS nrDNA sequence data confirm divergent lineages and paraphyly. Mycol. Res. 109 : 41-56.
31.Sun, J. D., Xiong, S. T., and Wang, P. 2009. Study on biological and cultivated characters of Cordyceps militaris SN3. J. Fungal Res. 7 : 148-152.
32.Sung, G. H., Hywel-Jones, N. L., Sung, J. M. Luangsa-Ard, J. J., Shrestha, B., and Spatafora, J. W. 2007. Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Stud. Mycol. 57 : 50-59.
33.Sung, J. M., Choi, Y. S., Shrestha, B., and Park, Y. J. 2002. Investigation on artificial fruiting of Cordyceps militaris. Kor. J. Mycol. 30 : 6-10.
34.Sung, J. M., Park, Y. J., Lee, J. O., Han, S. K., Lee, W. H., Choi, S. K., and Shrestha, B. 2006. Effect of preservation periods and subcultures on fruiting body formation of Cordyceps militaris in vitro. Mycobiol. 34 : 196-199.
35.Tan, Q., Cai, T., Wei, J., Feng, A. P., Mao, W. J., and Bao, D. P. 2011. Molecular identification of mating type genes in asexual spores of Cordyceps militaris. ICMBMP7 2011 : 52-56.
36.Tong, Y. K., Kuang, T., Wu, Y. X., Zhang, Q. Y., and Ren, J. 1997. Comparison of components of Cordyceps mycelium and natural Cordyceps sinensis. Shi Pin Yan Jiu Yu Kai Fa 18 : 40-42.
37.Turgeon, B. G., and Yoder, O. C. 2000. Proposed nomenclature for mating type genes of filamentous ascomycetes. Fungal Genet. Biol. 31 : 1-5.
38.Wang, L, Zhang, W. M., Hu, B., Chen, Y. Q., and Qu, L. H. 2008. Genetic variation of Cordyceps militaris and its allies based on phylogenetic analysis of rDNA ITS sequence data. Fungal Diversity. 31 : 147-155.
39.Wang, X. Y., Rong, Y. W., Xu, L., Zhang, N. S., Li, C. R., and Fan, M. Z. 2010. Electrophoretic karyotype analysis of Paecilomyces militaris, the anamorph of Cordyceps militaris. J. Anhui Agric. Univ. 37 : 716-719.
40.Wen, T. C., Li, M. F., Kang, J. C., and He, J. 2012. A molecular genetic study on fruiting-body formation of Cordyceps militaris. Afr. J. Microbiol. Res. 6 : 5215-5221.
41.Wong, Y. Y., Moon, A., Duffin, R., Barthet-Barateig, A., Meijer, H. A., Clemens, M. J., and De Moor, C. H. 2010. Cordycepin inhibits protein synthesis and cell adhesion through effects on signal transduction. J. Biol. Chem. 285 : 2610-2621.
42.Wu, F. Y., Yan, H., Ma, X. A., Jia, J. Q., Zhang, G. Z., Guo, X. J., and Gui, Z. Z. 2011. Structural characterization and antioxidant activity of purified polysaccharide from cultured Cordyceps militaris. Afr. J. Microbiol. Res. 5 : 2743-2751.
43.Yokoyama, E., Yamagishi, K., and Hara, A. 2004. Development of a PCR-based mating-type assay for Clavicipitaceae. FEMS Microbiol. Lett. 237 : 205-212.
44.Yokoyama, E., Yamagishi, K., and Hara, A. 2003. Structures of the mating-type loci of Cordyceps takaomontana. Appl. Environ. Microbiol. 69 : 5019-5022.
45.Yoo, H. S., Shin, J. W., Cho, J. H., Son, C. G., Lee, Y. W., Park, S. Y., and Cho, C. K. 2004. Effects of Cordyceps militaris extract on angiogenesis and tumor growth. Acta Pharm. Sinica 25 : 657-665.
46.Yu, H. M., Wang, B. S., Huang, S. C., and Duh, P. D. 2006. Comparison of preventive effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J. Agaric. Food Chem. 54 : 3132-3138.
47.Zheng, P., Xia, Y. L., Xiao, G. H., Xiong, C. H., Hu, X., Zhang, S. W., Zheng, H. J., Huang, Y., Zhou, Y., Wang, S. Y., Zhao, G. P., Liu, X. Z., St Leger, R. J., and Wang, C. S. 2011. Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional Chinese medicine. Genome Biol. 12 : R116
48.Zhou, X. W., Gong, Z. H., Su, Y., Lin, J., and Tang, K. X. 2009. Cordycepin fungi: natural products, pharmacological functions and developmental products. J. Pharm. Pharmacol. 61 : 279-291.
2.Choi, I. Y., Choi, J. S., Lee, W. H., Yu, Y. J., Joung, G. T., Ju, I. O. and Choi, Y. K. 1999. The condition of production of artificial fruiting body of Cordyceps militaris. Kor. J. Mycol. 27 : 243-248.
3.Choi, Y. S., Kim, H. K., Lee, B. J., and Kim, Y. G. 2009. Characteristics and breeding of a new variety Cordyceps militaris 'Yedang 3'. J. Mushroom Sci. Production 7 : 182-186.
4.Das, S. K., Masuda, M., Sakurai, A., and Sakakibara, M. 2010. Medicinal uses of the mushroom Cordyceps militaris: Current state and prospects. Fitoterapia 81 : 961-968.
5.De Julian-Ortiz, J. V., Galvez, J., Munoz-Collado, C., Garcia-Domenech, R., and Gimeno-Cardona, C. 1999. Virtual combinatorial syntheses and computational screening of new potential anti-herpes compounds. J. Med. Chem. 42 : 3308-3314.
6.Dong, J. Z., Lei, C., Ai, X. R., and Wang, Y. 2012. Selenium enrichment on Cordyceps militaris Link and analysis on its main active components. Appl. Biochem. Biotechnol. 166 : 1215-1224.
7.Du, A. L., Zhang, X., and Zhang, H. Z. 2010. A new high cordycepin Cordyceps militaris cultivar 'Haizhou 1'. Acta Hortic. Sin. 37 : 1373-1374.
8.Fan, L., Pan, H., Scoccol, A. T., Pandey, A., and Soccol, C. R. 2006. Advances in mushroom research in the last decade. Food Technol. Biotechnol. 44 : 303-311.
9.Gao, X. H. 2008. Mating system of Cordyceps militaris. Acta Edulis Fungi 15 : 6-10.
10.Gui, Z. Z. and Zhu, Y. H. 2008. Advance on cultivation, bioactive compound and pharmacological mechanism of Cordyceps militaris. Sci. Seri 34 : 178-184.
11.Guo, H. P. and Yang, Z. M. 1999. Progress in research of pharmacological of Cordyceps sinensis. Traditional Herbal Drugs 30 : 231-233.
12.Hung, L. T., Keawsompong, S., Hanh, V. T., Sivichai, S., and Hywel-Jones, N. L. 2009. Effect of temperature on cordycepin production in Cordyceps militaris. Thai J. Agaric. Sci. 42 : 219-225.
13.Jiang, X. L. and Sun, Y. 1999. The determination of active components in various Cordyceps militaris strains. Acta Edulia Fungi 6 : 47-50.
14.Jiang, Y. and Yao, Y. J. 2003. Anamorphic fungi related to Cordyceps sinensis. Mycosystema 22 : 161-176.
15.Li, M. F. 2007. Molecular biology studies on the different phenotypes of fruiting-body forming of Cordyceps militaris. Master Thesis. Guizhou Univ., Guiyang, China.
16.Liang, Z. Q. 2001. current situation and ponderation of Cordyceps Fr. research and exploitation in China. Acta Edulis Fungi 8 : 53-62.
17.Liu, X. M., Cheng, Y. H., and Tian, D. 1999. Progress in the research of pharmacological Cordyceps in China. Natl. Product Res. Development 11 : 87-91.
18.Liu Z. Y. 1999. Studies on relationship between Cordyceps spp. and their anamorphs. PhD dissertation. Huazhong Agric. Univ., China.
19.Ma, T. Feng, Y., Wu, X. P., Zhang, Y. H., Ma, Y., and Wang, Z. L. 2007. Primary investigation of a host insect of Cordyceps militaris and analysis of its main ingredients. For. Res. 20 : 63-67.
20.Mao, X. B., Eksriwong, T., Chauvatcharin, S., and Zhong, J. J. 2005. Optimization of carbon source and C:N ratio for cordycepin production by submerged cultivation of medicinal mushroom Cordyceps militaris. Process Biochem. 40 : 1667-1672.
21.Mao, X. B. and Zhong, J. J. 2004. Hyperproduction of cordycepin by two-stage dissolved oxygen control in submerged cultivation of medicinal mushroom Cordyceps militaris in bioreactors. Biotechnol. Prog. 20 : 1408-1413.
22.Mao, X. B. and Zhong, J. J. 2006. Significant effect of NH4+ on cordycepin production by submerged cultivation of medicinal mushroom Cordyceps militaris. Enzyme Microbiol. Technol. 38 : 343-350.
23.Masuda, M., Urabe, E., Honda, H., Sakurai, A., and Sakakibara, M. 2007. Enhanced production of cordycepin by surface culture using the medicinal mushroom Cordyceps militaris. Enzyme Microbiol. Technol. 40 : 1199-1205.
24.Oh, S. W., Kim, S. H., Song, H. N., and Han, D. S. 2003. Comparative chemical compositions of four kinds of Tochkaso. Kor. J. Food Sci. Technol. 35 : 15-22.
25.Park, J. P., Kim, S. W., Hwang, H. J., and Yun, J. W. 2001. Optimization of submerged culture conditions for the mycelial growth and exo-biopolymer production by Cordyceps militaris. Lett. Appl. Microbiol. 33 : 76-81.
26.Patel, K. J. and Ingalhalli, R. S. 2013. Cordyceps militaris (L.: Fr.) Link An important medicinal mushroom. J. Pharm. Phytochem. 2 : 315-319.
27.Sato, H. and Shimazu, M. 2002. Stroma production for Cordyceps militaris (Clavicipitales: Clavicipitaceae) by injection of hyphal bodies to alternative host insects. Appl. Entomol. Zool. 37 : 85-92.
28.Shih, I. L., Tsai, K. L., and Hsieh, C. Y. 2006. Effects of culture conditions on the mycelial growth and bioactive metabolite production in submerged culture of Cordyceps militaris. Biochem. Eng. J. 33 : 193-201.
29.Shrestha, B., Lee, W. H., Han, S. K, and Sung, J. M. 2006. Observations on some of the mycelial growth and pigmentation characteristics of Cordyceps militaris isolates. Mycobiol. 34 : 83-91.
30.Stensrud, Ø, Hywel-Jones, N. L., and Schumacher, T. 2005. Towards a phylogenetic classification of Cordyceps: ITS nrDNA sequence data confirm divergent lineages and paraphyly. Mycol. Res. 109 : 41-56.
31.Sun, J. D., Xiong, S. T., and Wang, P. 2009. Study on biological and cultivated characters of Cordyceps militaris SN3. J. Fungal Res. 7 : 148-152.
32.Sung, G. H., Hywel-Jones, N. L., Sung, J. M. Luangsa-Ard, J. J., Shrestha, B., and Spatafora, J. W. 2007. Phylogenetic classification of Cordyceps and the clavicipitaceous fungi. Stud. Mycol. 57 : 50-59.
33.Sung, J. M., Choi, Y. S., Shrestha, B., and Park, Y. J. 2002. Investigation on artificial fruiting of Cordyceps militaris. Kor. J. Mycol. 30 : 6-10.
34.Sung, J. M., Park, Y. J., Lee, J. O., Han, S. K., Lee, W. H., Choi, S. K., and Shrestha, B. 2006. Effect of preservation periods and subcultures on fruiting body formation of Cordyceps militaris in vitro. Mycobiol. 34 : 196-199.
35.Tan, Q., Cai, T., Wei, J., Feng, A. P., Mao, W. J., and Bao, D. P. 2011. Molecular identification of mating type genes in asexual spores of Cordyceps militaris. ICMBMP7 2011 : 52-56.
36.Tong, Y. K., Kuang, T., Wu, Y. X., Zhang, Q. Y., and Ren, J. 1997. Comparison of components of Cordyceps mycelium and natural Cordyceps sinensis. Shi Pin Yan Jiu Yu Kai Fa 18 : 40-42.
37.Turgeon, B. G., and Yoder, O. C. 2000. Proposed nomenclature for mating type genes of filamentous ascomycetes. Fungal Genet. Biol. 31 : 1-5.
38.Wang, L, Zhang, W. M., Hu, B., Chen, Y. Q., and Qu, L. H. 2008. Genetic variation of Cordyceps militaris and its allies based on phylogenetic analysis of rDNA ITS sequence data. Fungal Diversity. 31 : 147-155.
39.Wang, X. Y., Rong, Y. W., Xu, L., Zhang, N. S., Li, C. R., and Fan, M. Z. 2010. Electrophoretic karyotype analysis of Paecilomyces militaris, the anamorph of Cordyceps militaris. J. Anhui Agric. Univ. 37 : 716-719.
40.Wen, T. C., Li, M. F., Kang, J. C., and He, J. 2012. A molecular genetic study on fruiting-body formation of Cordyceps militaris. Afr. J. Microbiol. Res. 6 : 5215-5221.
41.Wong, Y. Y., Moon, A., Duffin, R., Barthet-Barateig, A., Meijer, H. A., Clemens, M. J., and De Moor, C. H. 2010. Cordycepin inhibits protein synthesis and cell adhesion through effects on signal transduction. J. Biol. Chem. 285 : 2610-2621.
42.Wu, F. Y., Yan, H., Ma, X. A., Jia, J. Q., Zhang, G. Z., Guo, X. J., and Gui, Z. Z. 2011. Structural characterization and antioxidant activity of purified polysaccharide from cultured Cordyceps militaris. Afr. J. Microbiol. Res. 5 : 2743-2751.
43.Yokoyama, E., Yamagishi, K., and Hara, A. 2004. Development of a PCR-based mating-type assay for Clavicipitaceae. FEMS Microbiol. Lett. 237 : 205-212.
44.Yokoyama, E., Yamagishi, K., and Hara, A. 2003. Structures of the mating-type loci of Cordyceps takaomontana. Appl. Environ. Microbiol. 69 : 5019-5022.
45.Yoo, H. S., Shin, J. W., Cho, J. H., Son, C. G., Lee, Y. W., Park, S. Y., and Cho, C. K. 2004. Effects of Cordyceps militaris extract on angiogenesis and tumor growth. Acta Pharm. Sinica 25 : 657-665.
46.Yu, H. M., Wang, B. S., Huang, S. C., and Duh, P. D. 2006. Comparison of preventive effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J. Agaric. Food Chem. 54 : 3132-3138.
47.Zheng, P., Xia, Y. L., Xiao, G. H., Xiong, C. H., Hu, X., Zhang, S. W., Zheng, H. J., Huang, Y., Zhou, Y., Wang, S. Y., Zhao, G. P., Liu, X. Z., St Leger, R. J., and Wang, C. S. 2011. Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional Chinese medicine. Genome Biol. 12 : R116
48.Zhou, X. W., Gong, Z. H., Su, Y., Lin, J., and Tang, K. X. 2009. Cordycepin fungi: natural products, pharmacological functions and developmental products. J. Pharm. Pharmacol. 61 : 279-291.