The effects of natural substrates on the sporulation and viability of conidia and blastospores of Metarhizium anisopliae

Document Type : Research Paper

Authors

BCPP-1508-49 (R2)

Abstract

In this study the effects of natural grain-based media on the sporulation and germination of conidia and blastospores of Metarhizium anisopliae were investigated. Experiments were conducted with factorial arrangement in a completely randomized design with nine treatments and three replications. The obtained results showed that there was significant difference in the yield of conidia among different treatnents, two weeks after inoculation, (F= 4.66, df= 7, 16, p<0.01). Highest average yields were obtained as 3.1×108 and 3×108 conidia/g in rice flour and Sabouraud’s Dextrose Agar with 1% yeast extract, respectively. The lowest level of conidia yield was 4×106 conidia/g which obtained in the wheat bran medium. There was significant difference between blastospores yields four days after inoculation of the media, (F=­2.57, df= 8, 18, p<0.05). The highest and lowest average yield of blastospores were 6.1×108 and 4.5×108 blastospores/ml and were obtained in the wheat bran+rice bran+yeast extract and rice bran+yeast extract media, respectively. Analysis of variance of nutritional and chemical amendments which were added to blastospores media showed significant differences in the yield among different treatments (F=5.02, df=13, 29, p<0.01). The highest average yield of blastospores were 6.6×108, 6.1×108 and 5.7 ×108 blastospores/ml which were obtained in the rice bran+wheat bran+yeast extract+whey powder, rice bran+wheat bran+yeast extract, and wheat bran+rice bran+magnesium sulfate media, respectively. The lowest average yield of the blastospores was obtained in the media containing grape juice. Results also showed significant difference between the means of germination rate of conidia one, two and three month storage at 4°C and 25°C. The highest germination rate belonged to treatments stored in the refrigerator.

 

Keywords

References

Arcas, J.A., Diaz, B.M., & Lecuona, R.E. 1999. Bioinsecticidal activity of conidia and dry mycelium preparations of two isolate of Beauveria bassiana against the sugarcane borer, Diatraea saccharalis. Journal of Biotechnology, 67: 151-158.
Assaf, A., Escobar, F. & de la Torre, M. 2009. Culture medium improvement for Isaria fumosorosea submerged conidia production. Biochemical Engineering Journal, 47: 87-92.
Bena-Molaei, P., Talaei-Hassanloui, R., Askary, H., and Kharazi-Pakdel, A. 2011. Study on potential of some solid natural substances in production of Beauveria bassiana (Ascomycota, Cordycipitaceae) conidia. Journal of Entomological Society of Iran, 30(2): 1-15. (In Farsi).
Dorta, B., Bosch, A., Arcas, J.A., & Ertola, R.J. 1990. High level of sporulation of Metarhizium anisopliae in a medium containing by-products. Applied Microbiology Biotechnology, 33: 712-715.
EL Damir, M. 2006. Effect of growing media and water volume on conidial production of Beauveria bassiana and Metarhizium anisopliae. Journal of Biological Sciences, 6(2): 269-274.
Fargus, J., Smits, N., Vidal, C., Vega, F., & Mercadier, G. 2001. Effect of liquid culture media on morphology, growth, propagule production, and pathogenic activity of the hyphomycete, Metarhizium flavoviridae. Mycopathologica, 154: 127-138.
Goettel, M.S. & Inglis, G.D. 1997. Fungi: Hyphomycetes. pp. 213-249. In: Lacey L.A. (ed.), Manual of Techniques in Insect Pathology. Academic Press, San Diego, USA.
Farsi, M.J., Askary, H., Talebi Jahromi, KH., & Kharazi Pakdel, A. 2005. Effect of important ecological factors on blastospore production of Verticillium lecanii DAOM 198499 in liquid medium. Iranian, Journal of Agriculture Science, 36(1):109-119. (In Persian).
Feng, M.G., Liu, B.L., & Tzeng, Y.M. 2000. Verticillium lecanii spore production in solid-state and liquid-state fermentations. Bioprocess and Biosystems Engineering,23: 25-29.
Ibrahim, L., Butt, T., & Jenkinson, P. 2002. Effect of artificial culture media on germination, growth, virulence and surface properties of the enthomopathogenic Hyphomycete Metarhizium anisopliae. Mycological Research, 106: 705-715.
Issaly, N., Chauveau, H., Aglevor, F., Fargues, J., & Durand, A. 2005. Influence of nutrient, pH and dissolved oxygen on the production of Metarhizium flavoviride Mf189 blastospores in submerged batch culture. Process Biochemistry, 40: 1425-1431.
Jackson, M.A. 1997. Optimizing nutritional conditions for the liquid culture production of effective fungal biological control agents. Journal of Industrial Microbiology & Biotechnology, 19: 180-187.
Jackson, M.A., Erhan, S., & Poprawski, T.J. 2006. Influence of formulation additives on the desiccation tolerance and storage stability of blastospores of the entomopathogenic fungus Paecilomyces fumosoroseus (Deuteromycotina:Hyphomycetes).Biocontrol Science and Technology, 16(1): 61-75.
Jackson, M.A., Cliquet, S. & ItenL, B. 2003. Media and fermentation processes for the rapid production of high concentrations of stable blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus, Biocontrol Science and Technology, 13: 23-33.
Jenkins, N.E., & Prior, C. 1993. Growth and formation of true conidia by Metarhiziumflavoviride in a simple liquid medium. Mycological Research, 97(12): 1489-1494.
Jenkins, N.E., Heviefo, G., Langewald, J., Cherry, A.J., & Lomer, C.J. 1998. Development of mass production technology for aerial conidia for use as mycopesticides. Biocontrol News and Information, 19: 21-31.
Jenkins, N.E., Ali, B.S. & Moore, D. 2007. Mass production of entomopathogenic fungi for biological control of insect pest. pp. 287-293. In: Parker, B.L., Skinner, M., EL Bouhssini, M. & S. G. Kumair (eds), Sunn Pest Management, A Decade of Progress 1994-2004. ICARDA, Aleppo, Syria, Arab Society for Plant Protection.
Kassa, A., Brownbridge, M., Parker, B., Skinner, M, Gouli, V., Gouli, T., Guo, S.M., & Lee, F. 2007. Evaluation of whey powder as a basal media ingredient for mass production of Beauveria bassiana and Metarhizium anisopliae. Proceedings of the 40th Annual meeting of the society for invertebrate pathology and 1st international forum on the entomopathogenic nematodes and symbiotic bacteria, 12-16 Aug., Quebec City, Universite Laval, Canada, 97.
Mascarin, G.M., Alves, S.B., & Lopes, R. 2010. Culture media selection for mass production of Isaria fumosorosea and Isaria farinose. Brazilian Archives of Biology and Technology, 53(4): 753-761.
Ortiz-Urquiza, A., Keyhani, N.O., & Quesada-Moraga E. 2013. Culture conditions affect virulence and production of insect toxic proteins in the entomopathogenic fungus Metarhizium anisopliae. Biocontrol Science and Technology, 23(10): 1199-1212.
Pham,T.H., Kim, J.J., Kim, G.S., & Kim, K. 2009. Production of blastospores of entomopathogenic fungus Beauveria bassiana in a submerged batch culture. Mycobiology, 37(3): 218-224.
Safavi, S.A., Shah, F.A., Pakdel, A.K., Rasoulian, G.R., Bandani, A.R., & Butt, T.M. 2007. Effect of nutrition on growth and virulence of the entomopathogenic fungus Beauveria bassiana, FEMS Microbiology Letters, 270: 116-123.
Rangel, D.E.N., Alston, D.G., & Roberts, D.W. 2008. Effects of physical and nutritional stress conditions during mycelial growth on conidial germination speed, adhesion to host cuticle, and virulence of Metarhizium anisopliae, an entomopathogenic fungus. Mycological Research, 112: 1355-1361.
Roshandel, S., Talaei Hassanlouei, R., Askary, H., Allahyari, H. & Marzban, R. 2013. Effect of culture substrates on virulence of Metarhizium anisopliae conidia and blastospores against Sunn pest, Eurygaster integriceps. Iranian Journal of Plant Protection Science, 44(2): 225-234.
Shah, F., Wang, C.S., & Butt, T.M. 2005. Nutrition influences growth and virulence of the insect-pathogenic fungus Metarhizium anisopliae FEMS. MicrobiologyLetters, 251(2): 259-266.
Tlecuital-Beristain, S., Gonzalez, G.V., Diaz-Godinez, G., & Lovea, O. 2010. Medium selection and effect of higher Oxygen concentration pulses on Metarhizium anisopliae Var Lepidiotum conidial production and quality. Mycopathologia, 169: 367-394.
Thomas, K.C., Khachatourians, G., & Ingledew, W.M. 1987. Production and properties of Beauveria bassiana conidia cultivated in submerged culture. Canadian Journal of Microbiology, 33: 12-20.
Wraight, S.P., Jackson, M.A. & Kock, S.L. 2001. Production, stabilization and formulation of fungal biocontrol agents. pp. 253-287. In: Butt, T.M., Jackson, C.W. & Magan, N. (eds), Fungi as Biocontrol Agents: Progress, Problems and Potential. CABI publishing, Wallingford,
Wu, J.H., Ali, S., Huang, Z., Ren, S. X., & Cai, S.J. 2010. Media composition influences growth, enzyme activity and virulence of the entomopathogen Metarhizium anisopliae (Hypocreales: Clavicipitaceae). Pakistan Journal of Zoology, 42: 451-459.
Wyss, G.S., Charudattan, R., & DeValerio, J.T. 2001. Evaluation of agar and grain media for mass Production of conidia of Dactylaria higginsii. Plant Disease, 85: 1165-1170.

Zimmermann, G. 1993. The entomopathogenic fungus Metarhizium anisopliae and its potential as a biocontrol agent. Pesticide Science, 37(4): 375-379.

BioControl in Plant Protection
Volume 4, Issue 1 - Serial Number 7
January 2017
Pages 94-104

Files

Share

How to cite

Statistics