Floresta e Ambiente
Floresta e Ambiente
Original Article Conservation of Nature

Morphological and Molecular Identification of Entomopathogenic Fungi from Agricultural and Forestry Crops

Gustavo Angel Bich; María Lorena Castrillo; Fernando Luis Kramer; Laura Lidia Villalba; Pedro Darío Zapata

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Abstract A promising alternative for insect pest control in agriculture is the use of entomopathogenic fungi. The objectives of this work were to isolate and identify morphologically and molecularly strains of entomopathogenic fungi from crops of agricultural and forestry interest for the development of insect biocontrol technologies in Argentina. Three strains of Beauveria bassiana, and two strains of Metarhizium anisopliae were isolated and morphologically identified. Identity values greater than 99% with strains of B. bassiana and M. anisopliae were obtained in molecular databases. It was also confirmed by phylogenetic methods that the sequences obtained from these strains were positioned within the clade that contained the sequences of reference of B. bassiana and M. anisopliae respectively. These results deepen the knowledge of the mycobiota of Argentinian crops as a basis for the development of biotechnological products in the area of biological control.


Beauveria, Metarhizium, Microscopy, rDNA, Phylogeny


Becerra VV, Paredes C, Rojo CM, France AL. RAPD e ITS detectan variación molecular en poblaciones chilenas de Beauveria bassiana. Agricultura Técnica. 2007;67:115-25.

Bich GA, Castrillo ML, Villalba LL, Zapata PD. Isolation of the symbiotic fungus of Acromyrmex pubescens and phylogeny of Leucoagaricus gongylophorus from leaf-cutting ants. Saudi Journal of Biological Sciences. 2017;24:851-6.

Boucias DG, Tigano MS, Sosa DR, Glare TR, Inglis PW. Genotypic Properties of the Entomopathogenic Fungus Nomuraea rileyi. Biological Control Journal. 2000;19:124-38.

Coates B, Hellmich R, Lewis L. Allelic variation of a Beauveria bassiana (Ascomycota: Hypocreales) minisatellite is independent of host range and geographic origin. Genome. 2002;45:125-32.

De A, Bose R, Kumar A, Mozumdar S. Targeted Delivery of Pesticides Using Biodegradable Polymeric Nanoparticles. Springer Briefs in Molecular Science. 2014.

Destéfano RH, Destéfano SAL, Messias CL. Detection of Metarhizium anisopliae var. anisopliae within infected sugarcane borer Diatraea saccharalis (Lepidoptera, Pyralidae) using specific primers. Genetics and Molecular Biology. 2004;27:245-52.

Doberski JW, Tribe HT. Isolation of entomogenous fungi elm bark and soil with reference to ecology of Beauveria bassiana and Metarhizium anisopliae. British Mycological Society, Transactions. 1980;74:95-100.

Driver F, Milner RJ, Trueman WH. A taxonomic revision of Metarhizium based on a phylogenetic analysis of rDNA sequence data. Mycological Research. 2000;104:134-50.

Erler F, Ozgur A. Potential of two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae (Coleoptera: Scarabaeidae), as biological control agents against the June beetle. Journal of Insect Science. 2015;15:1-6.

Figueroa L, Varela A, Corredor D. Evaluación de sustratos naturales para la propagación masiva del hongo entomopatógeno Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes). Revista de Investigación. 2007;1(1):127-31.

Folgarait P, Marfetán J, Cafaro M. Growth and Conidiation Response of Escovopsis weberi (Ascomycota: Hypocreales) Against the Fungal Cultivar of Acromyrmex lundii (Hymenoptera: Formicidae). Environmental Entomology. 2011;40:342-9.

Hibbett DS, Ohman A, Glotzer D, Nuhn M, Kirk P, Nilsson RH. Progress in molecular and morphological taxon discovery in Fungi and options for formal classification of environmental sequences. Fungal Biology Reviews. 2011;25(1):38-47.

Humber R. Entomopathogenic fungal identification. 2005.

Humber R. Identification of entomopathogenic fungi. Manual of Techniques in Invertebrate Pathology. 2012.

Hyde K, Abd-Elsalam K, Cai L. Morphology: still essential in a molecular world. 2010;114:439-51.

Izurieta CA. Estrategia forestal de la provincia de misiones instrumentos para el diseño de políticas. 2000.

Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012;28:1647-9.

Kim K-H, Kabir E, Jahan SA. Exposure to pesticides and the associated human health effects. Science of the Total Environment. 2016;575:525-35.

Kindermann J, El-Ayouti Y, Samuels GJ, Kubicek CP. Phylogeny of the Genus Trichoderma Based on Sequence Analysis of the Internal Transcribed Spacer Region 1 of the rDNA Cluster. Fungal Genetics and Biology. 1998;24:298-309.

Kirk PM, Cannon PF, Minter DW, Stalpers JA. Dictionary of the Fungi. 2008.

Lacey LA, Solter LF. Initial handling and diagnosis of diseased invertebrates. Manual of Techniques in Invertebrate Pathology. 2012.

Lecuona RE. Microorganismos patógenos empleados en el control microbiano de insectos plaga. 1996.

Massoia E, Chebez J, Bosso A. Los mamíferos silvestres de la provincia de Misiones, Argentina. 2006.

Pantou MP, Mavridou A, Typas MA. IGS sequence variation, group-I introns and the complete nuclear ribosomal DNA of the entomopathogenic fungus Metarhizium: Excellent tools for isolate detection and phylogenetic analysis. Fungal Genetics and Biology. 2003;38:159-74.

Paz A, González M, Crawford A. Código de barras de la vida: Introducción y perspectiva. Acta Biológica Colombiana. 2011;16:161-76.

Piontelli LE. Manual de microhongos filamentosos communes I. 2015.

Posadas J, Comerio R, Mini J, Lecuona R. A novel dodine-free selective medium based on the use of cetyl trimethyl ammonium bromide (CTAB) to isolate Beauveria bassiana, Metarhizium anisopliae sensu lato and Paecilomyces lilacinus from soil. Mycologia. 2012;104:974-80.

Punja ZK, Utkhede RS. Using fungi and yeasts to manage vegetable crop diseases. Trends in Biotechnology. 2003;21:400-8.

Rehner SA, Buckley E. A Beauveria phylogeny inferred from nuclear ITS and EF1-a sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia. 2005;97:84-98.

Rehner SA, Minnis AM, Sung GH, Luangsa-ard JJ, Devotto L, Humber RA. Phylogeny and systematics of the anamorphic, entomopathogenic genus Beauveria. Mycologia. 2011;103:1055-73.

Samson RA, Evans HC, Latgé JP. Atlas of entomopathogenic fungi. 1988.

Sandoval B.C. Manual Técnico Manejo integrado de Enfermedades en cultivos hidropónicos. 2004.

Schapovaloff ME, Alves LF, Angeli UM, López-Lastra CC. Ocurrencia natural de hongos entomopatógenos en suelos cultivados con yerba mate (Ilex paraguariensis St. Hil.) en Misiones, Argentina. Revista Argentina de Microbiología. 2015;47:138-42.

Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA. Fungal Barcoding Consortium. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proceedings of the National Academy of Sciences. 2012;109:6241-6.

Shenoy BD, Jeewon R, Hyde KD. Impact of DNA sequence-data on the taxonomy of anamorphic fungi. Fungal Diversity. 2007;26:1-54.

Solé M. Caracterización morfológica y molecular de hongos queratinofílicos: el orden Onygenales.. 2002.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution. 2013;30:2725-9.

Teixera R, Faria M. Pequeno manual sobre fungos entomopatogênicos. 2010.

Tigano MS, Gomes CM, Sobral BWS. Genetic Variability among Brazilian Isolates of the Entomopathogenic Fungus Metarhizium anisopliae. Journal of Invertebrate Pathology. 1995;65:206-10.

Toju H, Tanabe AS, Yamamoto S, Sato H. High-Coverage ITS Primers for the DNA-Based Identification of Ascomycetes and Basidiomycetes in Environmental Samples. PlosOne. 2012;7.

Weber E. Densidad básica de madera de Pinus taeda L. Marion de diferentes edades, Misiones. Floresta. 2005;35:487-94.

White TJ, Bruns TD, Lee SB, Taylor JW., Innis MA, Gelfand DH, Sninsky JJ, White TJ. Analysis of phylogenetic relationships by amplification and direct sequencing of ribosomal genes. 1990.

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