Bioinformatic Tools in the Analysis of Determinants of Pathogenicity and Ecology of Entomopathogenic Fungi Used as Microbial Insecticides in Crop Protection

  • Uma Devi Koduru
  • Sandhya Galidevara
  • Annette Reineke
  • Akbar Ali Khan Pathan


Insect pathogenic fungi have a huge potential as microbial components of biopesticides which serve as benign components in plant protection. The infection cycle of these fungi is well known. Realising their potential and scope to improve their utility in phytomedicine, extensive work on the molecular biology of pathogenesis has been done in the past decade. Wet bench techniques like gene isolation, cloning and characterisation and gene knockout experiments to transcriptomics techniques like cDNA-AFLP, microarray, qPCR, cDNA, EST and SSH library construction, as well as whole genome sequencing and analysis of data with a suite of bioinformatic tools and pipelines integrated with several biological databases, were done to understand the process/processes involved at each stage of the infection cycle of the insect pathogenic fungi. These are in particular adherence of spores to the insect cuticle, factors that aid in coping with the physical stress conditions in the surrounding environment, formation of an infection peg, penetrance into the insect, factors that abet in overcoming insect defence systems and growth in the insect, production of toxic secondary metabolites that lead to insect death and surfacing out from the insect cadaver as well as sporulating to iterate the infection cycle on yet another insect. The picture that emerged is detailed in this chapter. The genes/proteins involved and the analyses that aided in their identification are described. Environmental genomics through multitag 454 pyrosequencing of rRNA sequence reads in deciphering the effect of the inundative application of an entomopathogenic fungus on the native soil fungal diversity is described. The chapter highlights the bioinformatics-bolstered investigation of the factors that influence the affectivity of insect pathogenic fungi as microbial biopesticides.


Entomopathogenic fungi Pathogenicity genes Fungal diversity Pyrosequencing In silico tools and pipelines Databases 



The authors are grateful to DST-DFG and DST-DAAD PPP programmes for financial support in carrying out collaborative research on entomopathogenic fungi. Mrs. Sandhya is thankful to CSIR, New Delhi, for a senior research fellowship.


  1. Altschul SF, Gish W, Miller W et al (1990) Basic local alignment search tool. J Mol Biol 215:403–410PubMedGoogle Scholar
  2. Amiri-Besheli B, Khambay S, Cameron S et al (2000) Inter-and intra-specific variation in destruxin production by insect pathogenic Metarhizium spp. and its significance to pathogenesis. Mycol Res 104:447–4452Google Scholar
  3. Ashelford KE, Chuzhanova NA, Fry JC et al (2005) At least 1 in 20 16S rRNA sequence records currently held in public repositories is estimated to contain substantial anomalies. Appl Environ Microbiol 71:7724–7736PubMedCentralPubMedGoogle Scholar
  4. Bagga S, Hu G, Screen SE, Leger RJ (2004) Reconstruction the diversification of subtilisins in the pathogenic fungus Metarhizium anisopliae. Gene 324:159–169PubMedGoogle Scholar
  5. Baldwin TK, Winnenburg R, Urban M et al (2006) The pathogen-host interactions database (PHI-base) provides insights into generic and novel themes of pathogenicity. Mol Plant Microbe Interact 19:1451–1462PubMedGoogle Scholar
  6. Baratto CM, Dutra V, Boldo JT et al (2006) Isolation characterization and transcriptional analysis of the chitinase chi2 gene (DQ011663) from the biocontrol fungus Metarhizium anisopliae var anisopliae. Curr Microbiol 53:217–221PubMedGoogle Scholar
  7. Billich A, Zocher RJ (1987) Enzymatic synthesis of cyclosporin A. J Biol Chem 262:17258–17259PubMedGoogle Scholar
  8. Bingle LEH, Simpson TJ, Lazarus CM (1999) Ketosynthase domain probes identify two subclasses of fungal polyketide synthase genes. Fungal Gen Biol 26:209–223Google Scholar
  9. Bisset J (1983) Notes on Tolypocladium and related genera. Can J Bot 61:1311–1329Google Scholar
  10. Boucias DG, Pendland JC, Latge JP (1988) Nonspecific factors involved in attachment of entomopathogenic deuteromycetes to host insect cuticle. Appl Environ Microbiol 54:1795–1805PubMedCentralPubMedGoogle Scholar
  11. Buée M, Reich M, Murat C et al (2009) 454 pyrosequencing analyses of forest soils reveal an unexpectedly high fungal diversity. New Phytol 184:449–456PubMedGoogle Scholar
  12. Butt TM, Goettel MS (2000) Bioassays of entomogenous fungi. In: Navon A, Ascher KRS (eds) Bioassays of entomopathogenic microbes and nematodes. CABI Publishing, Wallingford, pp 141–195Google Scholar
  13. Butt TM, Ibrahim L, Ball BY et al (1994) Pathogenicity of the entomogenous fungi Metarhizium anisopliae and Beauveria bassiana against crucifer pests and the honey bee. Biocontrol Sci Technol 4:207–214Google Scholar
  14. Butt TM, Jackson C, Magan N (2001) Production, stabilization and formulation of fungal biocontrol agents. In: Butt TM, Jackson C, Magan N (eds) Fungi as biocontrol agents: progress, problems and potential. CABI Publishing, Wallingford, pp 1–8Google Scholar
  15. Butterrs JA, Devi KU, Mohan MC et al (2003) Screening for tolerance to Bavistin, a Benzimidazole fungicide containing methyl benzimidazol-2-yl carbamate (MBC) among strains of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin: sequence analysis of the Beta – tubulin gene to identify mutations conferring tolerance. Mycol Res 107:260–266Google Scholar
  16. Caporaso JG, Kuczynski J, Stombaugh J et al (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336PubMedCentralPubMedGoogle Scholar
  17. Carlos LM, Palmieri DA, Souza VQ et al (2008) SSR locator: tool for simple sequence repeat discovery integrated with primer design and PCR simulation. Int J Plant Genomics 4:363–374Google Scholar
  18. Castlebury LA, Sutherland JB, Tanne LA et al (1999) Use of a bioassay to evaluate the toxicity of beauvericin to bacteria. World J Microbiol Biotechnol 15:131–133Google Scholar
  19. Cerenius L, Thornqvist PO, Vey A et al (1990) The effect of the fungal toxin destruxin E on isolated crayfish hemocytes. J Insect Physiol 36:785–789Google Scholar
  20. Chao A (1984) Non-parametric estimation of the number of classes in a population. Scand J Stat 11:265–270Google Scholar
  21. Chao A, Lee SM (1992) Estimating the number of classes via sample coverage. J Am Stat Assoc 87:210–217Google Scholar
  22. Charnley AK (1989) Mechanisms of fungal pathogenesis in insects. In: Whipps JM, Lumsden RD (eds) Biotechnology of fungi for improving plant growth. Oxford University Press, London, pp 86–125Google Scholar
  23. Charnley AK (2003) Fungal pathogens of insects: cuticle degrading enzymes and toxins. Adv Bot Res 40:241–321Google Scholar
  24. Charnley AK, Leger RJ (1991) The role of cuticle-degrading enzymes in fungal pathogenesis in insects. In: Cole RT, Hoch HE (eds) Fungal spore disease initiation in plants and animals. Plenum Press, New York/London, pp 267–287Google Scholar
  25. Chu M, Mierzwa R, Truumees I et al (1993) 2 novel diketopiperazines isolated from the fungus Tolypocladium sp. Tetrahedron Lett 34:7537–7540Google Scholar
  26. Clarkson JM, Charnley AK (1996) New insights into mechanisms homology modeling and protein engineering strategy of subtilases of fungal pathogenesis in insects. Trends Microbio 4:197–203Google Scholar
  27. Claros MG, Vincens P (1996) Computational method to predict mitochondrial proteins and their target sequences. Eur J Bio 241:779–786Google Scholar
  28. Clowee RK (2006) Estimates: statistical estimation of species richness and shared species from samples.version8, user guide and application published at
  29. Cole JR, Chai B, Farris RJ et al (2007) The ribosomal database project (RDP-II): introducing my RDP space and quality controlled public data. Nucleic Acids Res 35:D169–D172PubMedCentralPubMedGoogle Scholar
  30. Cook RJ (2000) Advances in plant health management in the 20th century. Annu Rev Phytopathol 38:95–116PubMedGoogle Scholar
  31. De Faria MR, Wraight SP (2007) Mycoinsecticides and mycoacaricides: a comprehensive list with worldwide coverage and international classification of formulation types. Biol Control 43:237–256Google Scholar
  32. De Santis TZ, Hugenholtz P, Larsen N et al (2006) Greengenes, a chimera checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol 72:5069–5072Google Scholar
  33. Dias BA, Neves PMOJ, Furlaneto Maia L et al (2008) Cuticle-degrading proteases produced by the entomopathogenic fungus Beuaveria bassiana in the presence of coffee berry borer cuticle. Braz J Microbiol 39:301–306PubMedCentralPubMedGoogle Scholar
  34. Donatti AC (2008) Production and regulation of cuticle degrading proteases from Beauveria bassiana in the presence of Rhammatocerus schistocercoides cuticle. Curr Microbiol 56:256–260PubMedGoogle Scholar
  35. Doytchinova IA, Flower DR (2008) Bioinformatic approach for identifying parasite and fungal candidate subunit vaccines. Open Vaccine J 1:22–26Google Scholar
  36. Dumas C, Robert P, Pais M et al (1994) Insecticidal and cytotoxic effects of natural and hemi synthetic destruxins. Comp Biochem Physiol 108:195–203Google Scholar
  37. Edgar RC (2004) MUSCLE: a multiple sequence alignment method with reduced time and space complexity. BMC Bioinform 5:113Google Scholar
  38. El-Sayed GN, Coudron TA, Ignoffo CM (1989) Chitinolytic activity and virulence associated with native and mutant isolates of an entomopathogenic fungus Nomuraea rileyi. J Invertebr Pathol 54:394–403Google Scholar
  39. El-Sayed GN, Ignof LTD et al (1993) Cuticular and non-cuticular substrate influence on expression of cuticle degrading enzymes non-cuticular substrate influence on expression of cuticle degrading enzymes. Mycopathologia 122:79–87Google Scholar
  40. Enkerli J, Widmer F, Gessler C et al (2001) Strain-specific microsatellite markers in the entomopathogenic fungus Beauveria brongniartii. Mycol Res 105:1079–1087Google Scholar
  41. Faircloth BC (2008) MSATCOMMANDER: detection of microsatellite repeat arrays and automated, locus-specific primer design. Mol Ecol Resour 8:92–94PubMedGoogle Scholar
  42. Falgueras J, Lara AJ, Fernandez-Pozo N et al (2010) SeqTrim: a high-throughput pipeline for pre-processing any type of sequence read. BMC Bioinformatics 11:38PubMedCentralPubMedGoogle Scholar
  43. Fang W, Leng B, Xiao Y et al (2005) Cloning of Beauveria bassiana chitinase gene Bbchitl and its application to improve fungal strain virulence. Appl Environ Microbiol 71:363–370PubMedCentralPubMedGoogle Scholar
  44. Fang W, Pava-Ripoll M, Wang SB et al (2009) Protein kinase A regulates production of virulence determinants by the entomopathogenic fungus Metarhizium anisopliae. Fungal Genet Biol 46:277–285PubMedGoogle Scholar
  45. Fang W, Fernandes EKK, Roberts DW et al (2010) A laccase exclusively expressed by Metarhizium anisopliae during isotropic growth is involved in pigmentation, tolerance to abiotic stresses and virulence. Fungal Genet Biol 42:602–607Google Scholar
  46. Freimoser FM, Screen S, Bagga S et al (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 149:239–247PubMedGoogle Scholar
  47. Freimoser EM, Hu G, Leger RJ (2005) Variation in gene expression patterns as the insect pathogen Metarhizium anisopliae adapts to different host cuticles or nutrient deprivation in vitro. Microbiology 151:361–371PubMedGoogle Scholar
  48. Gao Q, Jin K, Ying SH et al (2011) Genome sequencing and comparative transcriptomics of the model entomopathogenic fungi Metarhizium anisopliae and M. acridum. PLoS Genet 7:1–18Google Scholar
  49. Gibson DM, Krasnoff SB, Churchill ACL (2007) Searching for polyketides in insect pathogenic fungi. In: Rimando AM, Baerson SR (eds) Polyketides: biosynthesis, biological activity, and genetic engineering, vol 955. American Chemical Society, Washington, DC, pp 48–67Google Scholar
  50. Giongo A, Crabb DB, Davis-Richardson AG et al (2010) PANGEA: pipeline for analysis of next generation amplicons. ISME J 4:854–861Google Scholar
  51. Goettel MS, Hajek AE, Siegel JP et al (2001) Safety of fungal biocontrol agents. In: Butt TM, Jackson C, Magan N (eds) Fungi as biocontrol agents: progress, problems and potential. CABI Publishing, Wallingford, pp 347–376Google Scholar
  52. Guengerich FP (2001) Common and uncommon cytochrome P450 reactions related to metabolism and chemical toxicity. Chem Res Toxicol 14:611–650PubMedGoogle Scholar
  53. Gupta S, Roberts DW, Renwick JAA (1989) Preparative isolation of destruxins from Metarhizium anisopliae by high performance liquid chromatography. J Liq Chromatogr 12:383–395Google Scholar
  54. Gupta S, Krasnoff SB, Roberts DW et al (1992) Structure of efrapeptins from the fungus Tolypocladium niveum: peptide inhibitors of mitochondrial ATPase. J Org Chem 57:2306–2313Google Scholar
  55. Gupta S, Montillot C, Hwang YS (1994) Isolation of novel beauvericin analogues from the fungus Beauveria bassiana. J Nat Prod 58:733–738Google Scholar
  56. Hagn A, Pritsch K, Ludwig W et al (2003) Fungal diversity in agricultural soil under different farming management systems, with special reference to biocontrol strains of Trichoderma spp. Biol Fert Soils 38:236–244Google Scholar
  57. Hamill RL, Higgens CE, Boaz HE et al (1969) The structure of beauvericin, a new depsipeptide antibiotic toxic to Artemia salina. Tetrahedron Lett 49:4255–4258Google Scholar
  58. Hammer Ø, Harper DAT, Ryan PD (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontol Electron 4:9Google Scholar
  59. Hane JK, Lowe RG, Solomon PS et al (2007) Dothideomycete plant interactions illuminated by genome sequencing and EST analysis of the wheat pathogen Stagonospora nodorum. Plant Cell 19:3347–3368PubMedCentralPubMedGoogle Scholar
  60. Hartman AL, Riddle S, McPhillips T et al (2010) Introducing W.A.T.E.R.S: a workflow for the alignment, taxonomy, and ecology of ribosomal sequences. BMC Bioinformatics 11:317PubMedCentralPubMedGoogle Scholar
  61. Hirsch J, Galidevara S, Strohmeier S et al (2013) Effects on diversity of soil fungal community and fate of an artificially applied Beauveria bassiana strain assessed through 454 pyrosequencing. Microb Ecol. doi: 10.1007/s00248-013-0249-5 Google Scholar
  62. Holder DJ, Kirkland BH, Lewis MW et al (2007) Surface characteristics of the entomopathogenic fungus Beauveria (Cordyceps) bassiana. Microbiology 153:3448–3457PubMedGoogle Scholar
  63. Huber T, Faulkner G, Hugenholtz P (2004) Bellerophon: a programme to detect chimeric sequences in multiple sequence alignments. Bioinformatics 20:2317–2319PubMedGoogle Scholar
  64. Humber RA (1991) Fungal pathogens of aphids. In: Peters DC, Webster JA, Chlouber CS (eds) Proceedings on aphid plant interactions: populations to molecules still water. Okla State University, pp 45–56Google Scholar
  65. Hurlbert SH (1971) The non-concept of species diversity: a critique and alternative parameters. Ecology 52:577–586Google Scholar
  66. Huse SM, Huber JA, Morrison HG et al (2007) Accuracy and quality of massively parallel DNA pyrosequencing. Genome Biol 8:R143–R143.9PubMedCentralPubMedGoogle Scholar
  67. Huson DH, Mitra S, Ruscheweyh HJ et al (2011) Integrative analysis of environmental sequences using MEGAN4. Genome Res 21:1552–1560PubMedCentralPubMedGoogle Scholar
  68. Jackson CW, Heale JB, Hall RA (1985) Traits associated with virulence to the aphid Macrosiphoniella sanborni in eighteen isolates of Verticillium lecanii. Ann Appl Biol 106:39–48Google Scholar
  69. Jackson MA, Dunlap CA, Jaronski ST (2010) Ecological considerations in producing and formulating fungal entomopathogens for use in insect biocontrol. BioControl 55:129–145Google Scholar
  70. Kamper J, Kahmann R, Bölker M et al (2006) Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis. Nature 444:97–101PubMedGoogle Scholar
  71. Katoh K, Asimenos G, Toh H (2009) Multiple alignment of DNA sequences with MAFFT. Methods Mol Biol 537:39–64PubMedGoogle Scholar
  72. Khaldi N, Seifuddin FT, Turner G et al (2010) SMURF: genomic mapping of fungal secondary metabolite clusters. Fungal Genet Biol 47:736–741PubMedCentralPubMedGoogle Scholar
  73. Khan AAP (2006) A comparative transcriptomic analysis of the generalist entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin grown on different insect cuticles and synthetic medium through cDNA-AFLP display. Dissertation, Andhra University, Visakhapatnam, IndiaGoogle Scholar
  74. Khan AAP, Uma Devi K, Vogel H et al (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 Genet Biol 44:1231–1241Google Scholar
  75. Khan S, Guo L, Maimaiti Y et al (2012) Entomopathogenic fungi as microbial biocontrol agent. Mol Plant Breed 3:63–79Google Scholar
  76. Kim HK, Hoe HS, Suh D et al (1999) Gene structure and expression of the gene from Beauveria bassiana encoding bassiasinI, an insect cuticle-degrading serine protease. Biotechnol Lett 21:777–783Google Scholar
  77. Krasnoff SB, Reátegui RF, Wagenaar MM et al (2004) Cicadapeptins I and II: new Aib-containing peptides from the entomopathogenic fungus Cordyceps heteropoda. J Nat Prod 68:50–55Google Scholar
  78. Kulkarni RD, Kelkar HS, Dean RA (2003) An eight-cysteine-containing CFEM domain unique to a group of fungal membrane proteins. Trends Biochem Sci 28:118–121PubMedGoogle Scholar
  79. Kumar S, Carlsen T, Mevik BH et al (2011) CLOTU: an online application for processing and clustering of 454 amplicon reads into OTUs followed by taxonomic annotation. BMC Bioinformatics 12:1–9Google Scholar
  80. Lacey LA, Goettel M (1995) Current development in microbial control of insect pests and prospects for the early 21st century. Entomophaga 40:3–27Google Scholar
  81. Lacey LA, Frutos R, Kaya HK et al (2001) Insect pathogens as biological control agents: Do they have a future? Biol Control 21:230–248Google Scholar
  82. Lafon A, Han KH, Seo JA et al (2006) G-protein and cAMP mediated signaling in aspergilli: a genomic perspective. Fungal Genet Biol 43:490–502PubMedGoogle Scholar
  83. Li Y, Kelly WG, Logsdon JM Jr et al (2004) Functional genomic analysis of the ADP-ribosylation factor family of GTPases: phylogeny among diverse eukaryotes and function in C. elegans. FASEB J 18:1834–1850PubMedGoogle Scholar
  84. Lin L, Fang W, Liao X et al (2011) The MrCYP52 Cytochrome P450 Monooxygenase Gene of Metarhizium robertsii Is Important for Utilizing Insect Epicuticular Hydrocarbons. PLoS ONE 6:e28984PubMedCentralPubMedGoogle Scholar
  85. Liu S, Peng G, Xia Y (2012) The adenylate cyclase gene MaAC is required for virulence and multi-stress tolerance of Metarhizium acridum. BMC Microbiol 12:163PubMedCentralPubMedGoogle Scholar
  86. Lozupone C, Knight R (2005) UniFrac: a new phylogenetic method for comparing microbial communities. Appl Environ Microbiol 71:8228–8235PubMedCentralPubMedGoogle Scholar
  87. Mahé S, Duhamel M, Le Calvez TL (2012) PHYMYCO-DB: a curated database for analyses of fungal diversity and evolution. PLoS ONE 7:e431117Google Scholar
  88. McCune B, Mefford MJ (2011) PC-ord. Multivariate analysis of ecological data, version 6.MjM Software, Gleneden Beach, Oregon, U.S.AGoogle Scholar
  89. Medema MH, Blin K, Cimermancic P et al (2011) antiSMASH rapid identification, annotation and analysis of secondary metabolite biosynthesis gene clusters in bacterial and fungal genome sequences. Nucleic Acids Res 39:W339–W346PubMedCentralPubMedGoogle Scholar
  90. Molnar I, Gibson DM, Krasnoff SB (2010) Secondary metabolites from entomopathogenic Hypocrealean fungi. Nat Prod Rep 27:1241–1275PubMedGoogle Scholar
  91. Needleman SB, Wunsch CD (1970) A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol 48:443–453PubMedGoogle Scholar
  92. Nelson DR (1999) Cytochrome P450 and the individuality of species. Arch Biochem Biophys 369:1–10PubMedGoogle Scholar
  93. Oksanen J, Kindt R, Legendre P et al (2007) vegan: Community Ecology Package. R package version 1.8-8. Online at:
  94. Ortiz de Montellano PR (2005) Cytochrome P450: structure, mechanism, and biochemistry, 3rd edn. Kluwer Academic/Plenum Press, New York, p 689Google Scholar
  95. Pandey RV, Nolte V, Schlotterer C (2010) CANGS: a user-friendly utility for processing and analyzing 454 GS-FLX data in biodiversity studies. BMC Res Notes 3:3PubMedCentralPubMedGoogle Scholar
  96. Pedrini N, Zhang S, Juarez MP et al (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 156:2549–2557PubMedGoogle Scholar
  97. Petrosino JF, Highlander S, Luna RA et al (2009) Metagenomic pyrosequencing and microbial identification. Clin Chem 55:856–866PubMedCentralPubMedGoogle Scholar
  98. Pollard A, Wyn Jones KG (1979) Enzyme activities in concentrate solutions of glycine-betaine and other solutes. Planta 144:291–298PubMedGoogle Scholar
  99. Quast C, Pruesse E, Yilmaz P et al (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:D590–D596PubMedCentralPubMedGoogle Scholar
  100. Quince C, Lanzen A, Curtis TP et al (2009) Accurate determination of microbial diversity from 454 pyrosequencing data. Nat Methods 6:639–641PubMedGoogle Scholar
  101. Rappleye CA, Goldman WE (2008) Fungal stealth technology. Trends Immunol 29:18–24PubMedGoogle Scholar
  102. Rawlings ND, Barrett AJ, Bateman A (2012) MEROPS: the database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Res 40:D343–D350PubMedCentralPubMedGoogle Scholar
  103. Rehner SA, Buckley E (2005) A Beauveria phylogeny inferred from nuclear 1 ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97:84–98PubMedGoogle Scholar
  104. Roberts DW, St Leger RJ (2004) Metarhizium spp., cosmopolitan insect pathogenic fungi: mycological aspects. Adv Appl Microbiol 54:1–70PubMedGoogle Scholar
  105. Samuels RI, Charnley AK, Reynolds SE (1988) The role of destruxins in the pathogenicity of 3strains of Metarhizium anisopliae for the tobacco hornworm Manduca sexta. Mycopathologia 104:51–58Google Scholar
  106. Scheepmaker JWA, Butt TM (2010) Natural and released inoculum levels of entomopathogenic fungal biocontrol agents in soil in relation to risk assessment and in accordance with EU regulations. Biocontrol Sci Technol 20:503–552Google Scholar
  107. Schloss PD, Handelsman J (2005) Introducing DOTUR, a computer programme for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71:1501–1506PubMedCentralPubMedGoogle Scholar
  108. Schloss PD, Handelsman J (2006) Introducing Tree Climber, a test to compare microbial community structures. Appl Environ Microbiol 72:2379–2384PubMedCentralPubMedGoogle Scholar
  109. Screen S, Bailey A, Charnley K et al (1997) Carbon regulation of the cuticle-degrading enzyme PR1 from Metarhizium anisopliae may involve a trans-acting DNA-binding protein CRR1, a functional equivalent of the Aspergillus nidulans CREA protein. Curr Genet 31:511–518PubMedGoogle Scholar
  110. Shah FA, Wang CS, Butt TM (2005) Nutrition influences growth and virulence of the insect-pathogenic fungus Metarhizium anisopliae. FEMS Microbiol Lett 251:259–266PubMedGoogle Scholar
  111. Singleton DR, Furlong MA, Rathbun SL et al (2001) Quantitative comparisons of 16S rRNA gene sequence libraries from environmental samples. Appl Environ Microbiol 67:4374–4376PubMedCentralPubMedGoogle Scholar
  112. Small CLN, Bidochka MJ (2005) Up-regulation of Pr1, a subtilisin-like protease, during conidiation in the insect pathogen Metarhizium anisopliae. Mycol Res 109:307–313PubMedGoogle Scholar
  113. Solomon PS, Tan KC, Sanchez P et al (2004) The disruption of a G alpha subunit sheds new light on the pathogenicity of Stagonospora nodorum on wheat. Mol Plant Microbe Interact 17:456–466PubMedGoogle Scholar
  114. St Leger RJ, Wang C (2009) Entomopathogenic fungi and the genomic era. In: Stock SP, Vandenberg J, Glazer I, Boemare N (eds) Insect pathogens: molecular approaches and techniques. CABI Publishing, Wallingford, pp 366–400Google Scholar
  115. St Leger RJ, Joshi L, Bidochka MJ et al (1995) Protein synthesis in Metarhizium anisopliae growing on host cuticle. Mycol Res 99:1034–1040Google Scholar
  116. St Leger RJ, Joshi L, Bidochka MJ et al (1996) Characterization and ultra structural localization of chitinases’ from Metarhizium anisopliae, Metarhizium. flavoviride and Beauveria bassiana during fungal invasion of host (Manduca sexta) cuticle. Appl Environ Microbiol 62:907–912Google Scholar
  117. St Leger RJ, Joshi L, Roberts DW (1997) Adaptation of proteases and carbohydrates of saprophytic, phytopathogenic and entomopathogenic fungi to the requirements of their ecological niches. Microbiology 143:1983–1992PubMedGoogle Scholar
  118. Sun Y, Cai Y, Liu L et al (2009) ESPRIT: estimating species richness using large collections of 16S rRNA pyrosequences. Nucleic Acids Res 37:e76PubMedCentralPubMedGoogle Scholar
  119. Thomas MB, Read AF (2007) Fungal bioinsecticide with a sting. Nat Biotechnol 25:1367–1368PubMedGoogle Scholar
  120. Uma Devi K, Padmavathi J, Uma Maheswara Rao C et al (2008) A study of host specificity in the entomopathogenic fungus Beauveria bassiana (Hypocreales, Clavicipitaceae). Biocontrol Sci Technol 18:975–989Google Scholar
  121. Uma Devi K, Reineke G, Sandhya G et al (2012) Pathogenicity genes in entomopathogenic fungi used as biopesticides. In: Gupta VK, Ayyachamy M (eds) Biotechnology of fungal genes. Science Publishers, Enfield, pp 343–367Google Scholar
  122. Vega FE, GoettelMS BM et al (2009) Fungal entomopathogens: new insights on their ecology. Fungal Ecol 2:149–159Google Scholar
  123. Vestergaard S, Gillespie AT, Butt TM et al (1995) Pathogenicity of the hyphomycete fungi Verticillium lecanii and Metarhizium anisopliae to the western flower thrips, Frankliniella occidentalis. Biocontrol Sci Technol 5:185–192Google Scholar
  124. Wang C, St Leger RJ (2005) Developmental and transcriptional responses to host and non host cuticles by the specific locus pathogen Metarhizium anisopliae var.acridum. Eukaryot Cell 4:937–947PubMedCentralPubMedGoogle Scholar
  125. Wang C, St Leger RJ (2006) A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses. Proc Natl Acad Sci 103:6647–6652PubMedCentralPubMedGoogle Scholar
  126. Wang C, St Leger RJ (2007a) The Metarhizium anisopliae perilipin homologMPL1regulates lipid metabolism, appressorial turgor pressure, and virulence. J Biol Chem 282:21110–21115PubMedGoogle Scholar
  127. Wang C, St Leger RJ (2007b) The MAD1 adhesin of Metarhizium anisopliae links adhesion with blastospore production and virulence to insects and the MAD2 adhesin enables attachment to plants. Eukaryot Cell 6:808–816PubMedCentralPubMedGoogle Scholar
  128. Wang CS, Hu G, St Leger RJ (2005) Differential gene expression by Metarhizium anisopliae growing in root exudate and host (Manduca sexta) cuticle or hemolymph reveals mechanisms of physiological adaptation. Fungal Genet BioI 42:704–718Google Scholar
  129. Wang CS, Duan ZB, Leger RJ (2008) MOS1osmosensor of Metarhizium anisopliae is required for adaptation to insect host hemolymph. Eukaryot Cell 7:302–309PubMedCentralPubMedGoogle Scholar
  130. Webb CO, Ackerly DD, Kembel SW (2008) Phylocom: software for the analysis of phylogenetic community structure and trait evolution Bioinformatics 24:2098–2100Google Scholar
  131. Wraight SP, Jackson MA, de Kock SL (2001) Production, stabilization and formulation of fungal biocontrol agents. In: Butt TM, Jackson C, Magan N (eds) Fungi as biocontrol agents: progress, problems and potential. CABI Publishing, Wallingford, pp 253–287Google Scholar
  132. Wraight SP, Ramos ME, Williams JE et al (2003) Comparative virulence and host specificity of Beauveria bassiana isolates assayed against lepidopteran pests of vegetable crops. J Invertebr Pathol 103:186–199Google Scholar
  133. Wu D, Hartman A, Ward N et al (2008) An automated phylogenetic tree-based small subunit rRNA taxonomy and alignment pipeline (STAP). PLoS ONE 3(7):e2566PubMedCentralPubMedGoogle Scholar
  134. Wu GD, Lewis JD, Hoffmann C et al (2010) Sampling and pyrosequencing methods for characterizing bacterial communities in the human gut using 16S sequence tags. BMC Microbiol 30:206Google Scholar
  135. Xia Y, Clarkson JM, Charnley AK (2001) Acid phosphatases of Metarhizium anisopliae during infection of the tobacco hornworm Manduca sexta. Arch Microbiol 176:427–434PubMedGoogle Scholar
  136. Xiao G, Ying SH, Zheng P et al (2012) Genomic perspectives on the evolution of fungal entomopathogenicity in Beauveria bassiana. Sci Rep 2:483PubMedCentralPubMedGoogle Scholar
  137. Xu Y, Orozco R, Wijeratne EM et al (2008) Biosynthesis of the cyclooligomer depsipeptide beauvericin, a virulence factor of the entomopathogenic fungus Beauveria bassiana. Chem Biol 15:898–907PubMedGoogle Scholar
  138. Xu Y, Orozco R, Kithsiri Wijeratne EM et al (2009) Biosynthesis of the cyclooligomer depsipeptide bassianolide, an insecticidal virulence factor of Beauveria bassiana. Fungal Genet Biol 46:53–364Google Scholar
  139. Xue QX, Wang J, Huang BF et al (2010) A new manganese superoxide dismutase identified from Beauveria bassiana enhances virulence and stress tolerance when over expressed in the fungal pathogen. Appl Microbiol Biotechnol 86:1543–1553Google Scholar
  140. Yu Y, Breitbart M, McNairnie P et al (2006) FastGroupII: a web-based bioinformatics platform for analyses of large 16S rDNA libraries. BMC Bioinformatics 7:57PubMedCentralPubMedGoogle Scholar
  141. Zhang W, Yueqing C, Yuxian X (2008) Cloning of the subtilisin Pr1A gene from a strain of locust specific fungus Metarhizium anisopliae and functional expression of the protein in Pichia pastoris. World J Microbiol Biotechnol 24:2481–2488Google Scholar
  142. Zhang Y, Zhao J, Fang W et al (2009) Mitogen-activated protein kinase hog1 in the entomopathogenic fungus Beauveria bassiana regulates environmental stress responses and virulence to insects. Appl Environ Microbiol 75:3787–3795PubMedCentralPubMedGoogle Scholar
  143. Zhang Y, Zhang J, Jiang X et al (2010) Requirement of a mitogen-activated protein kinase for appressorium formation and penetration of insect cuticle by the entomopathogenic fungus Beauveria bassiana. Appl Environ Microbiol 76:2262–2270PubMedCentralPubMedGoogle Scholar
  144. Zheng P, Xia Y, Xiao G et al (2011) Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional Chinese medicine. Genome Biol 12:R116PubMedCentralPubMedGoogle Scholar
  145. Zhu Y, Pan J, Qiu J et al (2008) Isolation and characterization of a chitinase gene from entomopathogenic fungus Verticillium lecanii. Appl Environ Microbiol 39:314–332Google Scholar

Copyright information

© Springer India 2014

Authors and Affiliations

  • Uma Devi Koduru
    • 1
  • Sandhya Galidevara
    • 1
  • Annette Reineke
    • 2
  • Akbar Ali Khan Pathan
    • 3
  1. 1.Department of BotanyAndhra UniversityVisakhapatnamIndia
  2. 2.Institute of PhytomedicineHochschule Geisenheim UniversityGeisenheimGermany
  3. 3.Department of Biochemistry, College of ScienceKing Saud UniversityRiyadhKingdom of Saudi Arabia

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