Study of Morphology, Mycelial Growth and Exopolysaccharides Production of Cordyceps sp. Selected from Several Farms in Central Region of Thailand

Authors

  • Samart Taikhao Biotechnology Innovation Program, Division of Science, Faculty of Science and Technology, Rajamangala University of Technology Suvarnabhumi, Mueang Nonthaburi, Nonthaburi 11000, Thailand.
  • Kanya Kokaew Microbiology Program, Division of Science, Faculty of Science and Technology, Rajamangala University of Technology Suvarnabhumi, Phra Nakhon Si Ayutthaya, Phra Nakhon Si Ayutthaya 13000, Thailand.
  • Dounghatai Rattanasatchatham Microbiology Program, Division of Science, Faculty of Science and Technology, Rajamangala University of Technology Suvarnabhumi, Phra Nakhon Si Ayutthaya, Phra Nakhon Si Ayutthaya 13000, Thailand.
  • Suntara Fueangfung Biotechnology Innovation Program, Division of Science, Faculty of Science and Technology, Rajamangala University of Technology Suvarnabhumi, Mueang Nonthaburi, Nonthaburi 11000, Thailand.

Keywords:

Cordyceps sp., mycelial growth, mycelial morphology

Abstract

Cordyceps militaris is an entomopathogenic fungus that can produce high levels of bioactive compounds such as cordycepin, adenosine, and polysaccharides. This research aimed to study mycelial morphology, growth, biomass, and exopolysaccharides production of Cordyceps sp. strains selected from several farms in central Thailand. Eleven samples of Cordyceps sp. were selected from six farms in Thailand. They were isolates CM-Sp01, CM-Sp02, CM-Sp03, CM-Sp04, CM-Sp05, CM-Sp06, CM-Sp07, CM-Sp08, CM-Sp09, CM-Sp10 and CM-Sp11. The hyphae morphology using the slide culture technique and microscopic examination were investigated. The results revealed that all isolates had septate hyphae. A round and an oval shape of conidia at the end of hyphae were also found. However, the size of conidia was different between Cordyceps sp. isolates. In addition, the study of morphology and density of mycelial colony of C. militaris isolates on PDA medium were determined. The results found that vegetative and aerial mycelium in all isolates were white. The isolate CM-Sp02 showed the highest mycelium density. Under determining the growth of mycelial colony by measuring colony diameter on PDA medium, isolate CM-Sp08 showed the highest growth rate (5.06 ± 0.65 mm/day) and length of diameter of mycelial colony (66.12 ± 1.45 mm) after 21 days of cultivation in PDA medium under temperature of 22 oC in dark condition. Mycelial biomass and exopolysaccharides in PDB medium were investigated. The result found that isolate CM-Sp08 showed the highest biomass production (55.20 ± 1.65 g/L) and the highest exopolysaccharide content (55.20 ± 1.65 g/L). This indicated that the isolate CM-Sp08 had high potential for growth, biomass production, and exopolysaccharide production.

References

Chen, B., Sun, Y., Luo, F. and Wang, C. 2020a. Bioactive metabolites and potential mycotoxins produced by Cordyceps fungi: A review of safety. Toxins 12(6): 410-422.

Chen, B.X., Wei, T., Xue, L.N., Zheng, Q.W., Ye, Z.W., Zou, Y., Yang, Y., Yun, F., Guo, L.Q. and Lin, J.F. 2020b. Transcriptome analysis reveals the flexibility of cordycepin network in Cordyceps militaris activated by L-Alanine addition. Frontiers in Microbiology 11: 577.

ChokeUmnuay, N. and Owatworakit, A. 2021. Effect of successive subculture of Cordyceps militaris on growth, metabolites production and stability of Rhf1 gene. Agriculture and Natural Resources 55(4): 537-546.

Dang, H.N., Wang, C.L. and Lay, H.L. 2018. Effect of nutrition, vitamin, grains, and temperature on the mycelium growth and antioxidant capacity of Cordyceps militaris (strains AG-1 and PSJ-1). Journal of Radiation Research and Applied Sciences 11: 130-138.

DuBois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A. and Smith, F. 1956. Colori- metric method for determination of sugars and related substances. Analytical Chemistry 28(3): 350-356.

Ha, S.Y., Jung, J.Y., Park, H.M. and Yang, J.K. 2022. Comparison of the metabolic profile of the mycelia and fruiting bodies of artificially cultured Cordyceps militaris. Journal of Mushrooms 20(1): 13-21.

Kontogiannatos, D., Koutrotsios, G., Xekalak, S. and Zervakis, G.I. 2021. Biomass and cordycepin production by the medicinal mushroom Cordyceps militaris-A review of various aspects and recent trends towards the exploitation of a valuable fungus. Journal of Fungi 7(11): 986.

Kunhorm, P., Chaicharoenaudomrung, N. and Noisa, P. 2019. Enrichment of cordycepin for cosmeceutical applications: culture systems and strategies. Applied Microbiology and Biotechnology 103(4): 1681-1691.

Lee, B.J., Lee, M.A., Kim, Y.G., Lee, K.W., Choi, Y.S., Lee, B.E. and Song, H.Y. 2013a. Cultural characteristics of Cordyceps militaris strain ‘Yedang 3’ on various media and nutritional conditions. Journal of Mushroom Science and Production 11(3): 124-130.

Lee, S.Y., Debnath, T., Kim, S.K. and Lim, B.O. 2013b. Anti-cancer effect and apoptosis induction of cordycepin through DR3 pathway in the human colonic cancer cell HT-29. Food and Chemical Toxicology 60: 439-447.

Lou, H.W., Lin, L.F., Guo, L.Q., Wang, X.W., Tian, S., Liu C., Zhao, Y. and Zhao, R. 2019b. Advances in research on Cordyceps militaris degeneration. Applied Microbiology and Biotechnology 103(19): 7835-7841.

Lou, H.W., Zhao, Y., Tang, H.B., Ye, Z.W., Wei, T., Lin, J.F. and Guo, L.Q. 2019a. Transcriptome analysis of Cordyceps militaris reveals genes associated with carotenoid synthesis and identification of the function of the cmtns Gene. Frontiers in. Microbiolology 10: 2105.

Maftoun, P., Malek, R., Abbas, M., Aziz, R. and Enshasy, H.E. 2013. Bioprocess for semi-industrial production of immunomodulatory polysaccharide pleuran by Pleurotus ostreatus in submerged culture. Journal of Scientific and Industrial Research 72(11): 655-662.

Nurmamat, E., Xiao, H., Zhang, Y. and Jiao, Z. 2018. Effects of different temperatures on the chemical structure and antitumor activities of polysaccharides from Cordyceps militaris. Polymers 10(4): 430.

Shih, I.L., Tsai, K.L. and Hsieh, C. 2007. Effects of culture conditions on the mycelial growth and bioactive metabolite production in submerged culture of Cordyceps militaris. Biochemical Engineering Journal 33(3): 193-201.

Shrestha, B., Han, S., Yoon, K. and Sung, J. 2005. Morphological characteristics of conidiogenesis in Cordyceps militaris. Mycobiology 33(2): 69-76.

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. Mycobiology 34(2): 83-91.

Singpoonga, N., Sang-on, B. and Chaiprasart, P. 2019. Effects of preservation method on fruiting body formation and cordycepin production of Cordyceps militaris culture. Agriculture and Natural Resources 53(2): 106-113.

Soltani, M., Al-Ali, M., Othman, N.Z., Malik, R., Elmarzugi, N., Aziz, R. and El-Enshasy, H.A. 2015. Medium composition effects on growth kinetic of Cordyceps militaris cells using agar plate method. Journal of Pharmacy and Biological Sciences 10(1): 79-82.

Taikhao, S., Fueangfung, S., Leasen, S. and Lorwongtragool, P. 2018. Study of cultivation substrate materials on growth and bioactive compounds production in Cordyceps militaris, pp. 434-443. In Watjanatepin, N., ed. The 3rd National Conferences Rajamangala University of Technology Suvarnabhumi. Rajamangala University of Technology Suvarnabhumi. (in Thai)

Wang, C.C., Wu, J.Y., Chang, C.Y., Yu, S.T. and Liu, Y.C. 2019. Enhanced exopolysaccharide production by Cordyceps militaris using repeated batch cultivation. Journal of Bioscience and Bioengineering 127(4): 499-505.

Wang, F., Song, X., Dong, X., Zhang, J. and Dong, C. 2017. DASH- type cryptochromes regulate fruiting body development and secondary metabolism differently than CmWC-1 in the fungus Cordyceps militaris. Applied Microbiology and Biotechnology 101(11): 4645-4657.

Wang, L., Xu, N., Zhang, J., Zhao, H., Lin, L., Jia, S. and Jia, L. 2015. Antihyperlipidemic and hepatoprotective activities of residue polysaccharide from Cordyceps militaris SU-12. Carbohydrate Polymers 131: 355-362.

Werapan, B., Nutaratat, P., Ariyaphuttarat, S. and Prathumpai, W. 2022. Cordycepin production by the potential fungal strains Cordyceps militaris BCC 2819 and Cordyceps cicadae BCC 19788 in submerged culture during batch and Fed-batch fermentation. African Journal of Biotechnology 21(10): 483-503.

Wijedasa, M.H. and Liyanapathirana, L.V.C. 2012. Evaluation of an alternative slide culture technique for the morphological identification of fungi species. Sri Lanka Journal of Infectious Diseases 2(2): 47-52.

Wu, H., Rao, Z.C., Cao, L., De Clercq, P. and Han, R.C. 2020. Infection of Ophiocordyceps sinensis fungus causes dramatic changes in the microbiota of its Thitarodes host. Frontiers in Microbiology 11: 577268.

Xia, Y., Luo, F., Shang, Y., Chen, P., Lu, Y. and Wang, C. 2017. Fungal cordycepin biosynthesis is coupled with the production of the safeguard molecule pentostatin. Cell Chemical Biology 24(12): 1479-1489.

Yin, J., Xin, X.D., Weng, Y.J., Li, S.H., Jia, J.Q. and Gui, Z.Z. 2018. Genotypic analysis of degenerative Cordyceps militaris cultured in the pupa of Bombyx mori. Entomological Research 48(3): 137-144.

Zhang, J., Wen, C., Duan, Y., Zhang, H. and Ma, H. 2019. Advance in Cordyceps militaris (Linn) Link polysaccharides: Isolation, structure, and bioactivities: A review. International Journal of Biological Macromolecules 132: 906-914.

Zhao, H., Lai, Q., Zhang, J., Huang, C. and Jia, L. 2018. Antioxidant and hypoglycemic effects of acidic-extractable polysaccharides from Cordyceps militaris on type 2 diabetes mice. Oxidative Medicine Cellular Longevity 2018: 9150807.

Zheng, P., Xia, Y., Xiao, G., Xiong, C., Hu, X., Zhang, S., Zheng, H., Huang, Y., Zhou, Y., Wang, S., Zhao, G., Liu, X., Leger, R.J.S. and Wang, C. 2011. Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional chinese medicine. Genome Biology 12(11): 1-21.

Published

2024-08-28

How to Cite

Taikhao, S. ., Kokaew, K., Rattanasatchatham, D., & Fueangfung, S. (2024). Study of Morphology, Mycelial Growth and Exopolysaccharides Production of Cordyceps sp. Selected from Several Farms in Central Region of Thailand . Recent Science and Technology, 16(3), 743–761. Retrieved from https://li01.tci-thaijo.org/index.php/rmutsvrj/article/view/254628