Journal of Crop Protection

Expression analyses of some Beauveria bassiana genes in response to cuticles of four different insects

10.48311/jcp.2015.1249
Volume 4, Issue 5
December 2015
Pages 675-690
Journal of Crop Protection

Authors

M. Kordi 1
N. Farrokhi 2
A. Masoudi 3
A. Derakhshan Shadmehri 3
S. Gharanjik 3

1 Biotechnology Research Department, Shahrood University, Shahrood, Iran.

2 Department of Biotechnology Engineering, Faculty of Energy Engineering and New Technologies, Shahid Beheshti University, Tehran, Iran.

3 Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahrood University, Sahrood, Iran.

Abstract
Beauveria spp. as entomopathogenic fungi have extremely diverse insect host ranges.Here, a comparative transcript analysis of a Bassiasin I (BSNI), a Dipeptidyl peptidase (DPP), a Cytochrom P450 monooxygenase (CYP52), a Mitogen activated protein kinase (MAPK), and a Neuronal calcium sensor 1 (NCS) in two isolates of Beauveria bassiana., with low and high virulence, were carried out. For this, induction media, containing isolated cuticles from four insects, and a check medium lacking any cuticular extract were prepared and isolates were allowed to grow on them. Interestingly, up-regulation of BSNI and DPP in induction media was noted in comparison to the check, following normalization with γ-ACTIN. CYP52 was up-regulated in synthetic medium in comparison to cuticular extract. MAPK expression was greater in most virulent isolate once compared with the low virulent isolate. The expression of MAPK was greater in the presence of Caliptamus italicus cuticles and synthetic medium as opposed to others. NCS expression remained on changed in all media tested and can be suggested as a reference gene in transcriptomics of Beauveria spp. genes in future.

Keywords

Bassiasin I
Dipeptidyl peptidase
Cytochrome P450 monooxygenase
Mitogen activated protein kinase
Neuronal calcium sensor
Abbott, W. S. 1925. A method of computing the effectiveness of an insecticide. Journal of Economic Entomology,18: 265-267.
Andersen, S. O. 1980. Cuticular sclerotization, In: Miller, T. A. (Eds.), Cuticle Techniques in Arthropods. Springer-Verlag, New York, pp. 185-217.
Bagga, S., Hu, G., Screen, S. E. and St. Leger, R. J. 2004. Reconstructing the diversification of subtilisins in the pathogenic fungus Metarhizium anisopliae. Gene, 324: 159-169.
Charnley, A. K. 1984. Physiological aspects of destructive pathogenesis in insects by fungi: a speculative review, In: Anderson, J. M., Rayner, A. D. M., Walton, D. W. H. (Eds.), Invertebrate–Microbial Interactions. British Mycological Society Symposium 6: Cambridge University Press, London, UK, pp. 229-270.
Dutra, V., Nakazato, L., Broetto, L., Silveira, I.S., Henning, M. V. and Schrank, A. 2004. Application of representational difference analysis to identify sequence tags expressed by Metarhizium anisopliae during the infection process of the tick Boophilus microplus cuticle. Research in Microbiology, 155: 245-251.
El Damir, M. 2006. Variation in germination, virulence and conidial production of single spore isolates of entomopathogenic fungi in response to environmental heterogeneity. Journal of Biological Sciences, 6: 305-315.
Fan, Y., Borovsky, D., Hawkings, C., Ortiz-Urquiza, A. and Keyhani, N. O. 2012a. Exploiting host molecules to augment mycoinsecticide virulence. Nature Biotechnology, 30: 35-37.
Fan, Y., Ortiz-urquiza, A., Kudia, R. A. and Keyhani, N. O. 2012b. A fungal homologue of neuronal calcium sensor-1, Bbcsa1, regulates extracellular acidification and contributes to virulence in the entomopathogenic fungus Beauveria bassiana. Microbiology, 158: 1843-1851.
Faria, M. R. and Wraight, S. P. 2007. Mycoinsecticides and mycoacaricides: a comprehensive list with worldwide coverage and international classification of formulation types. Biological Control, 43: 237-256.
Florez, A. M., Oviedo, A., Cardona, A., Herrera, M., Garcia, E., Restrepo, A. and Mc. Ewen, J. G. 2003. Molecular cloning and characterization of two hsp 70 homologous genes from the dimorphic fungus Paracoccidioides Brasiliensis. Biomedica, 23: 424-436.
Freimoser, F. M., Screen, S., Bagga, S., Hu, G. and St. Leger, R. J. 2003. Expressed sequence tag (EST) analysis of two subspecies of Metarhizium anisopliae reveals a plethora of secreted proteins with potential activity in insect hosts. Microbiology, 149: 239-247.
Freimoser, F. M., Hu, G. and St. Leger, R. J. 2005. Variation in gene expression patterns as the insect pathogen Metarhizium anisopliae adapts to different host cuticle or nutrient deprivation in vitro. Microbiology, 151: 361-371.
Goettel, M. S., St. Leger, R. J., Rizzo, N. W., Staples, R. C. and Roberts, D. W. 1989. Ultrastructural localization of a cuticle-degrading protease produced by the entomopathogenic fungus Metarhizium anisopliae during penetration of host (Manduca sexta) cuticle. Journal of General Microbiology, 135: 2233-2239.
Hackman, R. H. 1984. Cuticle: Biochemistry. Biology of the Integument, In: Bereiter-Hahn, J., Matolts, A. G. and Richards Berlin, K. S (Eds.), Springer-Verlag, New York, pp. 626-637.
Hegedus, D. D. and Khachatourians, G. G. 1995. The impact of biotechnology on hyphomycetous fungal insect biocontrol agents. Biotechnology Advances, 13: 455-490.
Hepburn, H. R. 1985. Structure of the integument. in: Comprehensive insect physiology, biochemistry and pharmacology, In: Kerkut, G. A. and Gilbert, L. I. (Eds.), Pergamon, Oxford, pp. 1-58.
IDT. 2015. Integrated DNA Technologies, Inc., Oligo Analyzer, http://eu.idtdna.com/ analyzer/Applic-ations/OligoAnalyzer.
Jin, K., Zhang, Y., Fang, W., Luo, Z., Zhou, Y. and Pei, Y. 2010. Carboxylate transporter gene JEN1 from the entomopathogenic fungus Beauveria bassiana is involved in conidiation and virulence. Applied and Environmental Microbiology, 76: 254-263.
Jin, K., Ming, Y. and Xia, Y. X. 2012. MaHog1, a hog1-type mitogen-activated protein kinase gene, contributes to stress tolerance and virulence of the entomopathogenic fungus Metarhizium acridum. Microbiology, 158: 2987-2996.
Kirkland, B. H., Westwood, G. S. and Keyhani, N. O. 2004. Pathogenicity of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to ixodidae tick species Dermacentor variabilis, Rhipicephalus sanguineus, and Ixodes scapularis. Journal of Medical Entomology, 41: 705-711.
McCoy, C. W., Samson, R. A. and Boucias, D. G. 1988. Entomogenous fungi, In: Ignoffo, C. M. and Mandava, N. B. (Eds.), Handbook of Natural Pesticides. Part A. Entomogenous Protozoa and Fungi. Microbial Pesticides, Inc, Boca Raton, FL: CRC Press, pp. 151-236.
Meyling, N. V. 2007. Method for isolation of entomopatogenic fungi from soil environment. In Department of Ecology (Frederiksberg Denmark University of Copenhagen). Http;// www.orgprints.org/11200. pp. 1-18
Neville, A. C. 1984. Cuticle: organisation. biology of the integument. In: Bereiter-Hahn, J., Matolts, A. G. and Richards Berlin, K. S., (Eds.), Springer-Verlag, New York, pp. 611-625.
Pathan, A. A. K., Uma Devi, K., Vogel, H. and Reineke, A. 2007. Analysis of differential gene expression in the generalist entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin grown on different insect cuticular extracts and synthetic medium through cDNA-AFLPs. Fungal Genetics and Biology, 44: 1231-1241.
Pedrini, N., Zhang, S., Juárez, M. P. and Keyhani, N. O. 2010. Molecular characterization and expression analysis of a suite of cytochrome P450 enzymes implicated in insect hydrocarbon degradation in the entomopathogenic fungus Beauveria bassiana. Microbiology, 156: 2549-2557.
Pendland, J. C., Hung, S. Y. and Boucias, D. G. 1993. Evasion of host defense by in vivo-produced protoplast-like cells of the insect mycopathogen Beauveria bassiana. Journal of bacteriology, 175: 5962-5969.
Plaza, G. A., Upchurch, R., Brigmon, R. L., Whitman, W. B. and Ulfig, K. 2004. Rapid DNA extraction for screening soil filamentous fungi using PCR amplification. Polish Journal of Environmental Studies, 13: 315-318.
Pontecorvo, G. 1953. The genetics of Aspergillus Nidulans. Advances in Genetics, 5: 141-239.
St. Leger, R. J., Bidochka, M. J., Roberts, D. W. 1994. Isoforms of the cuticle degrading Pr1 proteinase and production of a metalloproteinase by Metarhizium anisopliae. Archives of Biochemistry and Biophysics, 313: 1-7.
Sun, B. and Liu, X. 2008. Occurrence and diversity of insect-associated fungi in natural soils in China. Applied soil ecology,39: 100-108.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. and Kumar, S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood evolutionary distance and maximum parsimony methods. Molecular Biology and Evolution, 28: 2731-2739.
Tartar, A. and Boucias, D. G. 2004. A pilot-scale expressed sequence tag analysis of Beauveria bassiana gene expression reveals a tripeptidyl peptidase that is differentially expressed in vivo. Mycopathologia, 158: 201-209.
Vincent, J. F. V. and Shawky, N. A. F. 1978. The proteins of the urea-soluble fraction of locust intersegmental membrane. Insect Biochemistry, 8: 255-261.
Vincent, J. F. and Wegst, U. G. 2004. Design and mechanical properties of insect cuticle.Arthropod Structure & Development, 33: 187-199.
Wanchoo, A., Lewis, M. W. and Keyhani, N. O. 2009. Lectin mapping reveals stage-specific display of surface carbohydrates in in vitro and haemolymph-derived cells of the entomopathogenic fungus Beauveria bassiana. Microbiology, 155: 3121-3133.
Wang, C., Gang, H. and St. Leger, R. J. 2005. Differential gene expression by Metarhizium anisopliae growing in root exudate and host (Manduca sexta) cuticle or hemolymph reveals mechanisms of physiological adaptation. Fungal Genetics and Biology, 42: 704-718.
White, T. J., Bruns, T., Lee, S, Taylor, J. 1990. "Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols: A Guide to Methods and Applications." New York: Academic Press Inc: 315-322.
Zhang, Y., Zhao, J., Fang, W., Zhang, J., Luo, Z., Zhang, M., Fan, Y. and Pei, Y.2009. Mitogen-activated protein kinase Hog1 in the entomopathogenic fungus Beauveria bassiana regulates environmental stress responses and virulence to insects. Applied and Environmental Microbiology, 75: 3787-3795.
Zhang, S., Widemann, E., Bernard, G., Lesot, A., Pinot, F., Pedrini, N. and Keyhani, N. O. 2012. CYP52X1, representing new cytochrome P450 subfamily, displays fatty acid hydroxylase activity and contributes to virulence and growth on insect cuticular substrates in entomopathogenic fungus Beauveria bassiana. Journal of Biological Chemistry, 287: 13477-13486.
Zimmermann, G. 1986. The galleria bait method for detection of entomopathogenic fungi in soil. Journal of Applied Entomology-Zeitschrift Fur Angewandte Entomologie, 102: 213-215.

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