Email the author 
Alterations in electrophoresis patterns of Proteins, Isozymes and in vitro Protease activity under Thermal Stress in two isolates of Metarhizium anisopliae
Abstract
Two isolates of Metarhizium anisopliae (Metsch.) M33 (thermal tolerant) and M26 (thermal sensitive) were tested for biochemical alterations under thermal stress 37°C (M33T) to optimum temperature 25°C (M33T, M26) growth conditions. SDS-PAGE gel electrophoresis revealed variation in protein production resulting in the emergence of 9 heat shock proteins of different molecular mass by M33T under stress condition, those missing under optimum condition in M33C and M26. There was considerable trek in antioxidant enzymes, catalase and peroxidase by producing increased number of isoforms (2 and 3 isoforms respectively) on native PAGE gel with thermal stress (M33T) in comparison to optimum conditions (M 33C and M26). Other intra cellular enzymes, esterase and acid phosphatase depicted in loss of isoforms at 37°C when compared to M33C isolate. In vitro protease activity was in negative correlation with temperature stress, M33T isolate expressed the lowest protease activity throughout the incubation period of 6 days.
Keywords
Full Text:
PDFReferences
Barbaro, S. E., Trevors, J. T. and Inniss, W. E. 2001. Effects of low temperature, cold shock, and various carbon sources on esterase and lipase activities and exopolysaccharide production by a psychrotrophic Acinetobacter sp. Can. J. Microbiol. 47 (3) : 194–205.
Burger, M., Woods, R. G., McCarthy, C. and Beacham, I. R. 2000. Temperature regulation of protease in Pseudomonas fluorescens LS107d2 by an ECF sigma factor and a transmembrane activator. Microbiology. 146, 3149-3155
Butt, T. M., Jackson, C. and Magan, N. 2001. Introduction-fungal biological control agents:progress, problems and potential. In Fungal Biological Control Agents: Progress, Problems and Potential, pp. 1-8. Edited by T. M. Butt, C. Jackson and N. Magan. Oxford: CAB International.
Celik, E. and Calik, P. 2004. Bioprocess parameters and oxygen transfer characteristics in beta-lactamase production by Bacillus species, Biotechnol. Prog. 20, 491-499
Chen, K. Y. and Chen, Z. C. 2004. Heat shock proteins of thermophilic and thermotolerant fungi from Taiwan. Bot. Bull. Acad. Sin. 45: 247-257
Davidson, G., Phelps, K., Sunderland, K. D., Pell, J. K., Ball, B. V., Shaw, K. E. and Chandler, D. 2003. Study of temperature-growth interactions of entomopathogenic fungi with potential for control of Varroa destructor (Acari: Mesostigmata) using a nonlinear model of poikilotherm development. Journal of Applied Microbiology, 94: 816-825.
Devi, K.U., Sridevi, V., Mohan, Ch.M., Padmavathi, J., 2005. Effect of high temperature and water stress on in vitro germination and growth in isolates of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin. J. Invertebr. Pathol. 88, 181–189.
Duvetter T, Fraeye I, Sila D. N, Verlent I, Smout C, Clynen E, Schoofs L, Schols H, Hendrickx M and Van Loey A. 2006. Effect of temperature and high pressure on the activity and mode of action of fungal pectin methyl esterase. Biotechnol Prog. 22(5): 1313-20
Fernandes, E.K.K., Keyser, C.A., Chong, J.P., Rangel, D.E.N., Miller, M.P. and Roberts, D.W. 2010. Characterization of Metarhizium sps based on molecular analysis, heat tolerance and cold activity. Journal of Applied Microbiology. 108, 115-128.
Fernandes, E.K.K., Rangel, D.E.N., Moraes, A.M.L., Bittencourt, V.R.E.P. and Roberts, D.W. (2008) Cold activity of Beauveria and Metarhizium, and thermotolerance of Beauveria. J Invertebr Pathol 98, 69–78.
Hallsworth, J. E. and Magan, N. 1999. Water and temperature relations of growth of the entomogenous fungi Beauveria bassiana, Metarhizium anisopliae and Paecilomyces farinosus. Journal of Invertebrate Pathology. 74, 261-266
Hu, G. and St. Leger, R. J. 2002. Field studies using a recombinant mycoinsecticide (Metarhizium anisopliae) reveal that it is rhizosphere competent. Appl Environ Microbiol. 68, 6383-6387
Inglis, G. D., Goettel, M.S., Butt, T. M and Strasser, H. 2001. Use of hyphomycetous fungi for managing insect pests. In Fungi as Biocontrol Agents, pp. 23-69. Edited by T.M. Butt, C. Jackson and N. Magan. Wallingford: CAB International
Jernejc, K. and Cimerman, A. 2001. Morphological characteristics, extracellular and intra cellular proteins and enzyme patterns of five Aspergillus species. Food Technology and Biotechnology, 39 (4): 333-340.
Khachatourians, G. G., Qazi, S. S., and Reid, W. R. 2007. Biochemistry and molecular biology of entomopathogenic fungi. In: Brakhage, A. A., Zipfer, P. F. (Eds.), The Mycota VI Human and animal relationships. Springer-Verlag, Berlin Heidelberg, Germany, pp. 1-42, in press.
Li, J. and Feng, M.G. (2009) Intraspecific tolerance of Metarhizium anisopliae conidia to the upper thermal limits of summer with a description of a quantitative assay system. Mycol Res 113, 93–99.
Machwe, A., Senzik, A. M. and Kapoor, M., 2002. Induction profiles and properties of a novel stress induced peroxidase in N. crassa. Mycoscience 43:103-111.
Martins, V., Manfredini, V. and Benfato, M. S. 2005. High levels of Catalase in SOD mutants of Saccharomyces cervisiae in high aeration conditions. Braz. Journ. Microb. 36: 347-351
McKellar, R. C. and Cholette, H. 1987. Effect of temperature shifts on extracellular proteinase-specific mRNA pools in Pseudomonas fluorescens B52. Applied and Environmental Microbiology. 53, 1973-1976.
Moradas-Ferreira, P., Costa, V., Piper, P and Mager, W. 1996. The molecular defences against reactive oxygen species in yeast. Mol Microbiol. 19: 651-658
Noventa-Jordão, M. A., Couto, R. M., Goldman, M. H. S., Aguirre, J., Iyer, S., Caplan, A., Terenzi, H. F. and Goldman, G. H. 1999. Catalase activity is necessary for heat-shock recovery in Aspergillus nidulans germlings. Microbiology. 145: 3229-3234.
Oh, Y. S., Shih, I. L., Tzeng, Y. M. and Wang, S. L. 2000. Protease produced by Pseudomonas aeroginosa K-187 and its application in the deproteinization of shrimp and carb shell wastes. Enzyme Microb. Technol. 27, 3-10
Qazi, S. S. and Khachatourians, G. G. Hyderated conidia of Metarhizium anisopliae release a family of metalloproteases. Journal of Invertebrate Pathology (2007), doi: 10.1016/j.jip. 2006.12.002
Quesada-Moraga, E., Maranhao, E. A. A., Valverde-García, Santiago-Álvarez, C 2006. Selection of Beauveria bassiana isolates for control of the whiteflies Bemisia tabaci and Trialeurodes vaporariorum on the basis of their virulence, thermal requirements, and toxicogenic activity. Biological Control, 36: 274-287.
Rangel, D. E. N., Braga, G. U. L., Anderson, A. J. and Roberts, D. W. 2005. Variability in conidial thermotolerance of Metarhizium anisopliae isolates from different geographical origins. Journal of Invertebrate Pathology, 88, 116-125
Rao, Y. K., Lu, S. C., Liu, B. L. and Tzeng, Y. M. 2006. Enhanced production of an extracellular protease from Beauveria bassiana by optimization of cultivation processes. Biochemical Engineering Journal., 28, 27-66
Roberts, D.W. and St. Leger, R. J. 2004. Metarhizium spp., cosmopolitan insect-pathogenic fungi: mycological aspects. Adv Appl Microbiol 54, 1-70
Sadasivam, S. and Manickam, A.1991. Biochemical Methods.2nd Eds. New Age International (P) Ltd., Publishers, New Delhi.
Senczuk, A. M., Machwe, A. and Kapoor, M. 2003. High constitutive peroxidase activity and constitutive thermotolerance in Neurospora crassa. Mycoscience 44: 129-137.
Söderhäll, K. and Unestam, T. 1975. Properties of extracellular enzymes from Ampharomyces astaci and their relevance in the penetration process of crayfish cuticle. Physiol. Plant. 35: 140-146
Sujatha, K. 2009. Investigations on entomopathogenic fungal isolates Metarhizium anisopliae (metschinikoff) for detection of molecular markers. Submitted to department of Botany, Andhra University, Visakhapatnam.
Sumantha, A., Larroche, C. and Pandey, A. 2006. Microbiology and industrial biotechnology of food-grade proteases: A perspective. Food Technol. Biotechnol. 44 (2) 211-220
Tereshina, V. M. 2005. Thermotolerance in Fungi: The role of heat shock proteins and trehalose. Microbiology, 74, 247-257
Urtz, B. E. and Rice, W. C. 2000. Purification and characterization of a novel extracellular protease from Beauveria bassiana. Mycol. Res. 104, 180-186
Walstad, J. D., Anderson, R. F., Stambaugh, W. J. 1970. Effects of environmental conditions on two species of muscardine fungi (Beauveria bassiana and Metarhizium anisopliae). Journ. Invertebr. Pathol. 16, 221-226
Wyss, M., Pasamontes, L., Rémy, R., Kohler, J., Kusznir, E., Gadient, M., Müller and Van Loon, A. P. G. M. 1998. Comparison of the thermostability properties of three acid phosphotases from molds: Aspergillus fumigatus phytase, A. niger phytase, and A. niger pH 2.5 acid phosphatase. Applied and Environmental Microbiology. 4446-4451.
Copyright (c) 2014 Annals of Plant Sciences
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
