Abstract
Molecular biology methods have elucidated pathogenic processes in several biocontrol agents including one of the most commonly applied entomopathogenic fungi, Metarhizium anisopliae. In this article I will describe how a combination of EST and microarray approaches, gene disruption strategies, manipulation of gene expression and use of marker genes has: (1) identified and characterized genes involved in infection; (2) manipulated the genes of the pathogen to improve biocontrol performance; (3) allowed expression of a neurotoxin from the scorpion Androctonus australis; (4) allowed assessments of environmental risks posed by these modifications and (5) identified differences in genic constituents and gene expression that account for differences between strains.
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T. M. Butt, C. Jackson, and N. Magan, Introduction-fungal biological control agents: Progress, problems and potential, inFungal Biocontrol Agents: Progress, Problems and Potential, edited by T. M. Butt, C. Jackson, and N. Morgan (CAB International, Wallingford, UK, 2001), pp. 1–8.
R. P. Bateman, Controlled droplet application of mycoinsecticides: An environmentally friendly way to control locusts. Antenna 16, 6–13 (1992).
R. J. Milner, Selection and characterization of strains of Metarhizium anisopliae for control of soil insects in Australia, in Biological Control of Locusts and Grasshoppers, edited by C. J. Lomer and C. Prior (CAB International, Wallingford, UK, 1992), pp. 200–207.
C. Prior, Discovery and characterization of fungal pathogens for locust and grasshopper control, in Biological Control of Locusts and Grasshoppers, edited by C. J. Lomer and C. Prior (CAB International, Wallingford, UK, 1992), pp. 159–180.
A. Hajek, S. P. Wraight, and J. D. Vandenberg, Control of arthropods using pathogenic fungi, in Bio-exploitation of Filamentous Fungi, edited by S. P. Pointing and K. D. Hyde (Fungal Diversity Press, Hong Kong, 2001).
R. L. Harrison and B. C. Bonning, Genetic engineering of biocontrol agents for insects, in Biological and Biotechnological Control of Insect Pests, edited by J. E. Rechcigl and N. A. Rechcigl (Lewis Publishers, Boca RAton, FL, 1998), pp. 243–280.
S. P. Wraight, M. A. Jackson, and S. L. deKock, Production, stabilization and formulation of fungal bocontrol agents, inFungal biocontrol agents: Progress, problems and potential, edited by T. M. Butt, C. Jackson, and N. Morgan (CAB International, Wallingford, UK, 2001), pp. 253–287.
M. G. Klein, P. S. Grewal, and T. A. Jackson, Lawn, turf and grassland pests, in Field Manual of Techniques in Invertebrate Pathology, edited by L. A. Lacey and H. K. Kaya (Kluwer Academic Publishers, Dordrecht, 2000), pp. 681–706.
D. W. Roberts and R. J. St. Leger, Metarhizium spp. Cosmopolitan insect-pathogenic fungi: Mycological aspects, Adv. Appl. Microbiol. 54, 1–70 (2004).
M. A. Benjamin, E. Zhioua, and R. S. Ostfeld, Laboratory and field evaluation of the entomopathogenic fungus Metarhizium anisopliae (Deuteromycetes) for controlling questing adult Ixodes scapularis (Acari: Ixodidae), J. Med. Entomol. 39, 723–728 (2002).
M. Wright, A. K. Raina, and A. R. Lax, A strain of the fungus Metarhizium anisopliae for controlling subterranean termites, J. Econ. Entomol. 98, 1451–1458 (2005).
F. M. Freimoser, S. Screen, S. Bagga, G. Hu, and R. J. St. Leger, EST analysis of two subspecies of Metarhizium anisopliae reveals a plethora of secreted proteins with potential activity in insect hosts, Microbiology 149, 239–247 (2003).
F. M. Freimoser, S. Screen, G. Hu, and R. J. St. Leger, EST analysis of genes expressed by the zygomycete Conidiobolus coronatus during optimized secretion of proteins, Microbiology 149, 1893–1900 (2003).
F. M. Freimoser, G. Hu, and R. J. St. Leger, Variation in gene expression patterns as the insect pathogen Metarhizium anisopliae adapts to different host cuticles or nutrient deprivation in vitro, Microbiology 151, 361–371 (2005).
C. Wang, G. Hu, and R. J. St. Leger, Differential gene expression by Metarhizium anisopliae growing in root exudate and host (Manduca sexta) cuticle or hemolymph reveals mechanisms of physiological adaptation, Fungal Genet. Biol. 42, 704–718 (2005).
C. Wang, T. M. Butt, and R. J. St. Leger, Colony sectorization of Metarhizium anisopliae is a sign of ageing, Microbiology 151, 3223–3236 (2005).
C. Wang and R. J. St. Leger, A collagenous protective coat enables Metarhizium anisopliae to evade insect immunity, Proc. Natl. Acad. Sci. 103, 6647–6652 (2006).
R. J. St. Leger, L. Joshi, M. J. Bidochka, and D. W. Roberts, Construction of an improved mycoinsecticide over-expressing a toxic protease, Proc. Natl. Acad. Sci. 93, 6349–6354 (1996).
S. E. Screen, G. Hu, and R. J. St. Leger, Transformants of Metarhizium anisopliae sf. anisopliae overexpressing chitinase from Metarhizium anisopliae sf. acridum show early induction of native chitinase but are not altered in pathogenicity to Manduca sexta, J. Invertebr. Pathol. 78, 260–266 (2001).
A. Leclerque, H. Wan, A. Abschultz, S. Chen, G. V. Mitina, G. Zimmerman, and H. U. Scharer, Agrobacterium-mediated insertional mutagenesis (AIM) of the entomopathogenic fungus Beauveria bassiana, Curr. Genet. 45, 111–119 (2004).
R. J. St. Leger, B. May, L. Allee, D. C. Frank, D. W. Roberts, and R. C. Staples, Genetic differences in allozymes and in formation of infection structures among isolates of the entomopathogenic fungus Metarhizium anisopliae, J. Invertebr. Pathol. 60, 89–101 (1992).
M. J. Bidochka, R. J. St. Leger, and D. W. Roberts, Differentiation of species and strains of entomopathogenic fungi by random amplification of polymorphic DNA (RAPD), Curr. Genet. 25, 107–113 (1994).
G. H. Sung, J. W. Spatafora, R. Zare, K. T. Hodge, and W. Gams, A revision of Verticillium sect. Prostrata, II: Phylogenetic analysis of SSU and LSU nuclear rDNA sequences from anamorphs and teleomorphs of the Clavicipitaceae, Nova Hedwigia 72, 311–328 (2001).
F. Driver, R. J. Milner, and J. W. H. Trueman, A taxonomic revision of Metarhizium based on a phylogenetic analysis of rDNA sequence data, Mycol. Res. 104, 134–150 (2000).
J. Fargues, Adhesion of the fungal spore to the insect cuticle in relation to pathogenicity, in Infection Processes of Fungi, Conference Report, edited by D. W. Roberts and J. R. Aist (Rockefeller Foundation, 1984), pp. 90–110.
R. J. St. Leger, M. J. Bidochka, and D. W. Roberts, Germination triggers of Metarhizium anisopliae, Microbiology 140, 1651–1660 (1994).
B. Amiri-Besheli, B. Khambay, S. Cameron, M. L. Deadman, and T. M. Butt, Inter- and intra-specific variation in destruxin production by insect pathogenic Metarhizium spp., and its significance to pathogenesis, Mycol. Res. 104, 447–452 (2000).
T. M. Butt and M. Brownbridge, Increasing the efficacy of entomogenous fungi, in Enhancing Biocontrol Agents and Handling Risks, edited by M. Vurro, J. Gressel, T. Butts, G. Harman, A. Pilgeram, R. J. St. Leger, and D. Nuss (IOS Press, Amsterdam, The Netherlands, 2001), pp. 52–63.
G. D. Inglis, M. S. Goettel, T. M. Butt, and H. Strasser, Use of hyphomycetous fungi for managing insect pests, in Fungi as Biocontrol Agents, edited by T. M. Butt, C. Jackson, and N. Magan (CAB International, Wallingford, UK, 2001), pp. 23–69.
J. Drummond, J. B. Heale, and A. T. Gillespie, Germination and effect of reduced humidity on expression of pathogenicity in Verticillium lecani, Ann. Appl. Biol. 111, 193–201 (1987).
M. J. Bidochka, A. M. Kamp, T. M. Lavender, J. Dekoning, and J. N. A. De Croos, Habitat association in two genetic groups of the insect-pathogenic fungus Metarhizium anisopliae: Uncovering cryptic species? Appl. Environ. Microbiol. 67, 1335–1342 (2001).
D. E. N. Rangel, G. U. L. Braga, A. J. Anderson, and D. W. Robert, Influence of growth environment on tolerance to UV-B radiation, germination speed, and morphology of Metarhizium anisopliae var. acridum conidia, J. Invetebrate Pathol. 90, 55–58 (2005).
G. U. L. Braga, D. E. N. Rangel, S. D. Flint, A. J. Anderson, and D. W. Roberts, Conidial pigmentation is important to tolerance against solar-simulated radiation in the entomopathogenic fungus Metarhizium anisopliae, Photochem. Photobiol. 82, 418–422 (2006).
H. De Grotte, O. Douro-Kpindou, Z. Ouambama, C. Gbongboui, D. Müller, S. Attignon, and Chris Lomer, Assessing the feasibility of biological control of locusts and grasshoppers in West Africa: Incorporating the farmers’ perspective, Agricu. Human Values 18, 413–428 (2001).
Z. Long and D. M. Hunter, Laboratory and field trials of Green Guard® (Metarhizium anisopliase var. acridium) (Deuteromycotina: Hyphomycetes) against the oriental migratory locust (Locusta migratoria manilensis) (Orthoptera: Acrididae), China 14, 27–30 (2005).
E. J. Scholte, K. Ng’habi, J. Kihonda, W. Takken, K. Paaijmans, S. Abdulla, G. F. Killeen and B. G. J. Knols, An entomopathogenic fungus for control of adult African malaria mosquitoes, Science 308, 1641–1642 (2005).
S. Blanford, B. H. K. Chan, N. Jenkins, D. Sim, R. J. Turner, A. F. Read, and M. B. Thomas, Fungal pathogen reduces potential for malaria transmission, Science 308, 1638–1641 (2005).
A. Flores, Saving bees: Fungus found to attack Varroa mites, Agric. Res. Mag. 52, 18 (2004).
M. S. Goettel, and G. D. Inglis, Fungi: Hyphomycetes, in Manual of Techniques in Insect Pathology, edited by L. Lacey (Kluwer Academic Publishers, 1997), pp. 213–249.
M. J. Bidochka, Monitoring the fate of biocontrol fungi, inFungal Biocontrol Agents: Progress, Problems and Potential, edited by T. M. Butt, C. Jackson, and N. Morgan (CAB International, Wallingford, UK, 2001), pp. 193–218.
K. P. Barley, The configuration of the root system in relation to nutrient uptake, Adv. Agron. 22, 159–201 (1970).
T. F. Duda and S. R. Palumbi, Molecular genetics of ecological diversification: Duplication and rapid evolution of toxin genes of the venomous gastropod Conus, Proc. Natl. Acad. Sci. 96, 6820–6823 (1999).
G. E. Harman and B. G. G. Dozelli, Enhancing crop performance and pest resistance with genes from biocontrol agents, in Enhancing Biocontrol Agents and Handling Risks, edited by M. Vurro, J. Gressel, T. Butts, G. Harman, A. Pilgeram, R. St. Leger, and D. Nuss (IOS Press, Amsterdam, The Netherlands, 2001), pp. 114–125.
J. M. Whipps, Microbial interactions and biocontrol in the rhizosphere, J. Exp. Bot. 52, 487–511 (2001).
G. Hu and R. J. St. Leger, Field studies of a recombinant mycoinsecticide (Metarhizium anisopliae) reveal that it is rhizosphere competent. Appl. Environ. Microbiol. 68, 6383–6387 (2002).
R. J. St. Leger and S. Screen, Prospects for strain improvement of fungal pathogens of insects and weeds, in Fungal Biocontrol Agents: Progress, Problems and Potential, edited by T. M. Butt, C. Jackson, and N. Morgan (CAB International, Wallingford, UK, 2001), pp. 219–238.
R. J. St. Leger, M. J. Bidochka, and D. W. Roberts, Co-transformation of Metarhizium anisopliae by electroporation or the gene gun to produce stable GUS transformants, FEMS Microbiol. Lett. 131, 289–294 (1995).
B. R. Kerry, Rhizosphere interactions and the exploitation of microbial agents for the biological control of plant-parasitic nematodes, Annu. Rev. Phytopathol. 38, 423–441 (2000).
R. Baker, Diversity in biological control, Crop Protection 10, 85–94 (1991).
D. W. Roberts and A. E. Hajek, Entompathogenic fungi as bioinsecticides, in Frontiers in Industrial Mycology, edited by G. F. Leatham (Chapman & Hall, New York, 1992), pp. 144–159.
J. M. Barea, M. J. Pozo, R. Azcon, and C. Azcon-Aguilar, Microbial co-operation in the rhizosphere, J. Exp. Bot. 56, 1761–1778 (2005).
E. B. Nelson, Microbial dynamics and interactions in the spermosphere, Annu. Rev. Phytopathol. 42, 271–309 (2004).
P. Liang and A. B. Pardee, Analyzing differential gene expression in cancer, Nat. Rev. Cancer 3, 869–876 (2003).
Y. Ben-Shahar, A. Robichon, M. B. Sokolowski, and G. E. Robinson, Influence of gene action across different time scales on behavior, Science 296, 741–744 (2002).
W. Enard, P. Khaitovich, J. Klose, S. Zollner, F. Heissig, P. Giavalisco, K. Nielset-Struwe, E. Muchmore, A. Varki, R. Ravid, G. M. Doxiadis, R. E. Bontrop, and S. Paabo, Intra and inter specific variation in primate gene expression patterns, Science 296(5566), 233–235 (2002).
R. B. Brem, G. Yvert, R. Clinton, and L. Kruglyak, Genetic dissection of transcriptional regulation in budding yeast, Science 296, 752–755 (2002).
T. L. Ferea, D. Botstein, P. O. Brown, and R. F. Rosenzweig, Systematic changes in gene expression patterns following adaptive evolution in yeast, Proc. Natl. Acad. Sci. 96, 9721–9726 (1999).
J. P. Townsend, D. Cavalieri, and D. L. Hartl, Population genetic variation in genome-wide gene expression, Mol. Biol. Evol. 20, 955–963 (2003).
M. V. Olson, When less is more: Gene loss as an engine of evolutionary change, Am. J. Hum. Genet. 64, 18–23 (1999).
G. Hu and R. J. St. Leger, A phylogenomic approach to reconstructing the diversification of serine proteases in fungi, J. Evol. Biol. 17, 1204–1214 (2004).
R. H. Waterston, K. Lindblad-Toh, E. Birney, et al., Initial sequencing and comparative analysis of the mouse genome, Nature 420, 520–562 (2002).
D. G. Boucias and J. C. Pendland, Detection of protease inhibitors in the hemolymph of resistant Anticarsia gemmatalis inhibitory to the entomopathogenic fungus Nomuraea rileyi, Experientia 43, 336–3439 (1997).
S. Bagga, S. E. Screen, and R. J. St. Leger, Reconstructing the diversification of subtilisins in the pathogenic fungus Metarhizium anisopliae. Gene, 324, 159–169 (2004).
C. B. Michielse, P. J. Hooykaas, C. A. van den Hondel, and A. F. Ram, Grobacterium-mediated transformation as a tool for functional genomics in fungi, Curr. Genet. 48, 1–17 (2005).
L. Hoyer, The ALS gene family of Candida albicans, Trends Microbiol. 9, 176–180 (2001).
K. Al-Aidroos and D. W. Roberts, Mutants of Metarhizium anisopliae with increased virulence toward mosquito larvae, Can. J. Genet. Cytol. 20, 211–220 (1978).
A. K. Charnley, Fungal pathogens of insects: cuticle degrading enzymes and toxins, Adv. Bot. Res. 40, 241–321 (2003).
R. J. St. Leger, Biology and mechanisms of invasion of deuteromycete fungal pathogens, in: Parasites and Pathogens of Insects, edited by N. C. Beckage, S. N. Thompson, and B. A. Federici (Academic Press, New York, 1993), vol. 2, pp. 211–229.
R. J. St. Leger, R. M. Cooper, and A. K. Charnley, The effect of melanization of Manduca sexta cuticle on growth and infection by Metarhizium anisopliae, J. lnvertebr. Pathol. 52, 459–470 (1988).
C. E. Gongora, Transformacion de Beauveria bassiana cepa Bb9112 con les genes de la proteina verde fluorescente y la protease pr1A de M. anisopliae, Rev. Colomiana Entomol. 30, 1–5 (2004).
A. Flores, I. Chet, and A. Herrera Estrella, Improved biocontrol activity of Trichoderma harzianum by overexpression of the proteinase encoding gene, prb1, Curr. Genet. 31, 30–37 (1997).
J. Ahman, T. Johansson, M. Olsson, P. J. Punt, A. M. J. Cees, van den Hondel, and A. Tunlid, Improving the pathogenicity of a nematode-trapping fungus by genetic engineering of a subtilisin with nematotoxic activity, Appl. Environ. Microbiol. 68, 3408–3415 (2002).
A. C. Rath, The use of entomopathogenic fungi for control of termites, Biocont. Sci. Technol. 10, 563–581 (2000).
M. G. Villani, S. R. Krueger, P. C. Schroeder, F. Consolie, N. H. Console, L. M. Preston-Wisey, and D. W. Roberts, Soil application effects of Metarhizium anisopliae on Japanese beetle (Coleoptera: Scarabaeidae) behavior and survival in turfgrass microcosms, Environ. Entomol. 23, 502–513 (1994).
R. L. Samuels, S. E. Reynolds, and A. K. Charnley, Calcium-channel activation of insect muscle by destruxins, insecticidal compounds produced by the entomopathogenic fungus Metarhizium anisopliae. Comp. Biochem. Physiol. C 90, 403–412 (1988).
Vey, A., Matha, V., and Dumas, C. Effects of the peptide mycotoxin destruxin E on insect hemocytes and on dynamics and efficiency of the multicellular immune reaction, J. Invertebr. Pathol. 80, 177–187 (1988).
S. Pal, R. J. St. Leger, and L. P. Wu, Fungal peptide destruxin A plays a specific role in suppressing the innate immune response in Drosophila melanogaster, J. Biol. Chem. (2007) in press.
E. Zlotkin, Y. Fishman, M. Elazer, AaIT: From neurotoxin to insecticide, Biochemie 82, 869–881 (2000).
X. Sun, X. Chen, Z. Zhang, H. Wang, F. J. Bianchi, H. Peng, J. M. Vlak, and Z. Hu, Bollworm responses to release of genetically modified Helicoverpa armigera nucleopolyhedroviruses in cotton, J. Invertebre. Pathol. 81, 63–69 (2002).
M. Isaka, P. kittakoop, K. Kirtikra, N. Hywel-Jones, and Y. Thebtaranonth, Bioactive substances from insect pathogenic fungi, Acc. Chem. Res. 38, 813–823 (2005).
C. Wang and R. J. St. Leger, Developmental and transcriptional responses to host and non hostcuticles by the specific locust pathogen Metarhizium anisopliae sf. Acridum, Eukaryotic Cell 4, 937–947 (2005).
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Leger, R.J. (2007). METARHIZIUM ANISOPLIAE AS A MODEL FOR STUDYING BIOINSECTICIDAL HOST PATHOGEN INTERACTIONS. In: Vurro, M., Gressel, J. (eds) Novel Biotechnologies for Biocontrol Agent Enhancement and Management. NATO Security through Science Series. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-5799-1_9
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