Genetic analysis of tropical Beauveria and Metarhizium associated with sugar cane insects
DOI:
https://doi.org/10.15517/am.v30i1.32307Keywords:
Coleoptera, entomopathogenic fungi, Hemiptera, microsatellites, Saccharum officinarumAbstract
Introduction. Entomopathogenic fungi are used as biopesticides to control invertebrate pests in agriculture. The generation of genetic information and its geographic relationships are important for diversity studies and for the protection of biological resources. Objective. The aim of this study was to analyze the allelic diversity and the genetic and geographic relationships of several isolates of Beauveria and Metarhizium from Departamento de Investigación y Extensión de la Caña de Azúcar (DIECA) of Liga Agrícola Industrial de la Caña de Azúcar (LAICA). Materials and methods. This research was performed in DIECA and Universidad Nacional in 2014 and 2015. Fourteen and thirteen isolates of Beauveria and Metarhizium, respectively, were cultured from various Hymenoptera and Coleoptera insects of Costa Rica and Brazil. The isolates were analyzed using primer pairs that amplified simple sequence repeat (SSR) markers; twelve specific to Beauveria and thirteen specific to Metarhizium. Geographic and genetic correlation matrices were tested using a Mantel test. Results. All Beauveria primer pairs generated products, while only eight of the thirteen Metarhizium pairs generated products. Several sets of Beauveria primers were highly informative, but only one of the pairs was moderately informative in Metarhizium. Five Beauveria primer pairs generated unique genetic profiles in eight of the fourteen isolates (57%). Genetic relationships revealed two major clades among the Beauveria isolates, where one of these clades separated into two smaller groups. Metarhizium isolates were closely related, except for one (Mal12). A linear model explained 26% of the variability of the correlation between the genetic distances and the geographic distances found in Beauveria isolates, however, no relationships were found in Metarhizium isolates. Conclusion. A greater number of markers or different molecular marker technologies would be necessary to distinguish the isolates from the collection. The information in this work is useful for diversity studies and protection of intellectual property. Herein, we also consider search strategies to encounter diverse fungi.
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References
Aquino-De-Muro, M., S. Elliott, D. Moore, B.L. Parker, M. Skinner, W. Reid, and M. El-Bouhssini. 2005. Molecular characterisation of Beauveria bassiana isolates obtained from overwintering sites of Sunn Pests (Eurygaster and Aelia species). Mycol. Res. 109:294-306. doi:10.1017/S0953756204001832
Barbará, T., C. Palma-Silva, G.M. Paggi, F. Bered, M.F. Fay, and C. Lexer. 2007. Cross-species transfer of nuclear microsatellite markers: potential and limitations. Mol. Ecol. 16:3759-3767. doi:10.1111/j.1365-294X.2007.03439.x
Bautista-Glavez, A., J.F. Barrera, E. Payró-de-la-Cruz, S. Salgado-García, J. Gómez-Ruiz, and J.F. Gomez-Leyva. 2012. Genetic characterisation of Metarhizium anisopliae (Metchnikoff) Sorokin isolates from sugarcane fields and their pathogenicity against Aeneolamia postica (Walker) (Hemiptera: Cercopidae). Univ. Cienc. 28:217-229.
Becerra-Velásquez, V., M. Paredes-Cárcamo, C. Rojo-Meriño, A. France-Iglesias, and J. Franco-Durán. 2007. Intraspecific differentiation of Chilean isolates of the entomopathogenic fungi Metarhizium anisopliae var. anisopliae as revealed by RAPD, SSR and ITS markers. Genet. Mol. Biol. 30:89-99. doi:10.1590/S1415-47572007000100017
Bischoff, J.F., S.A. Rehner, and R.A. Humber. 2009. A multilocus phylogeny of the Metarhizium anisopliae lineage. Mycologia 101:512-530. doi:10.3852/07-202
Botstein, D., R.L. White, M. Skolnick, and R.W. Davis. 1980. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am. J. Hum. Genet. 32:314-331.
Bridge, P.D., C. Prior, J. Sagbohan, C.J. Lomer, M. Carey, and A. Buddie. 1997. Molecular characterization of isolates of Metarhizium from locusts and grasshoppers. Biodivers. Conserv. 6:177-189. doi:10.1023/A:1018387918686
Buntjer, J.B., and M. Otsen. 1999. Cross checker provides computer-assisted marker interpretation. J. Agric. Genomics 4:1-6.
Carracedo, A., and P. Sánchez-Diz. 2005. Forensic DNA typing technologies. In: A. Carracedo, editor, Forensic DNA typing protocols. Humana Press, Totowa, NJ, USA. p. 1-11.
Castle, A., D. Speranzini, N. Rghei, G. Alm, D. Rinker, and J. Bissett. 1998. Morphological and molecular identification of Trichoderma isolates on North American mushroom farms. Appl. Environ. Microb. 64:133-137.
Castrillo, L.A., J.D. Vandenberg, and S.P. Wraight. 2003. Strain-specific detection of introduced Beauveria bassiana in agricultural fields by use of sequence-characterized amplified region markers. J. Invertebr. Pathol. 82(2):75-83. doi:10.1016/S0022-2011(02)00190-8
Decroocq, V., M.G. Fave, L. Hagen, and L. Bordenave. 2003. Development and transferability of apricot and grape EST microsatellite markers across taxa. Theor. Appl. Genet. 106:912-922. doi:10.1007/s00122-002-1158-z
Fernandes, É.K., Á.M. Moraes, R.S. Pacheco, D.E. Rangel, M.P. Miller, V.R. Bittencourt, and D.W. Roberts. 2009. Genetic diversity among Brazilian isolates of Beauveria bassiana: Comparisons with non-Brazilian isolates and other Beauveria species. J. Appl. Microbiol. 107:760-774. doi:10.1111/j.1365-2672.2009.04258.x
FitzSimmons, N.N., C. Moritz, and S.S. Moore. 1995. Conservation and dynamics of microsatellite loci over 300 million years of marine turtle evolution. Mol. Biol. Evol. 12:432-440. doi:10.1093/oxfordjournals.molbev.a040218
Freed, S., F.L. Jin, and S.X. Ren. 2011. Determination of genetic variability among the isolates of Metarhizium anisopliae var. anisopliae from different geographical origins. World J. Microbiol. Biotechnol. 27:359-370. doi:10.1007/s11274-010-0466-8
Garrido-Jurado, I., M. Márquez, A. Ortiz-Urquiza, C. Santiago-Álvarez, E.A. Iturriaga, E. Quesada-Moraga, E. Monte, and R. Hermosa. 2011. Genetic analyses place most Spanish isolates of Beauveria bassiana in a molecular group with word-wide distribution. BMC Microbiol. 11:84. doi:10.1186/1471-2180-11-84
Ghikas, D.V., V.N. Kouvelis, and M.A. Typas. 2010. Phylogenetic and biogeographic implications inferred by mitochondrial intergenic region analyses and ITS1-5.8S-ITS2 of the entomopathogenic fungi Beauveria bassiana and B. brongniartii. BMC Microbiol. 10:174. doi:10.1186/1471-2180-10-174
Hadapad, A., A. Reineke, and C.P. Zebitz. 2006. Genetic variability among Beauveria brongniartii (Saccardo) Petch isolates from various geographical and host origins based on AFLP analysis. Mitt. Dtsch. Ges. Allg. Angew. Ent. 15:71-76.
Hernández-Domínguez, C., A.W. Guzmán-Franco, M.G. Carrillo-Benítez, R. Alatorre-Rosas, E. Rodríguez-Leyva, and J.A. Villanueva-Jiménez. 2016. Specific diversity of Metarhizium isolates infecting Aeneolamia spp. (Hemiptera: Cercopidae) in sugarcane plantations. Neotrop. Entomol. 45:80-87. doi:10.1007/s13744-015-0337-y
Hu, J.B., X.Y. Zhou, and J.W. Li. 2009. Development of novel chloroplast microsatellite markers for Cucumis from sequence database. Biol. Plant 53:793-796. doi:10.1007/s10535-009-0146-4
Hughes, W.O.H., L. Thomsen, J. Eilenberg, and J.J. Boomsma. 2004. Diversity of entomopathogenic fungi near leaf-cutting ant nests in a neotropical forest, with particular reference to Metarhizium anisopliae var. anisopliae. J. Invertebr. Pathol. 85:46-53. doi:10.1016/j.jip.2003.12.005
Keswani, C., K. Bisen, V. Singh, B.K. Sarma, and H.B. Singh. 2016. Formulation technology of biocontrol agents: Present status and future prospects. In: N.K. Arora et al., editors, Bioformulations: For sustainable agriculture. Springer, New Delhi, IND. p. 35-52.
Leal, C.M., D.J. Bertioli, T.M. Butt, J.H. Carder, P.R. Burrows, and J.F. Peberdy. 1997. Amplification and restriction endonuclease digestion of the Pr1 gene for the detection and characterization of Metarhizium strains. Mycol. Res. 101:257-265. doi:10.1017/S0953756296002560
Meyling, N.V., and J. Eilenberg. 2007. Ecology of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae in temperate agroecosystems: Potential for conservation biological control. Biol. Control 43:145-155. doi:10.1016/j.biocontrol.2007.07.007
Meyling, N.V., M. Lübeck, E.P. Buckley, J. Eilenberg, and S.A. Rehner. 2009. Community composition, host range and genetic structure of the fungal entomopathogen Beauveria in adjoining agricultural and seminatural habitats. Mol. Ecol. 18:1282-1293. doi:10.1111/j.1365-294X.2009.04095.x
Meyling, N.V., C. Pilz, S. Keller, F. Widmer, and J. Enkerli. 2012. Diversity of Beauveria spp. isolates from pollen beetles Meligethes aeneus in Switzerland. J. Invertebr. Pathol. 109:76-82. doi:10.1016/j.jip.2011.10.001
Nishi, O., K. Hasegawa, K. Iiyama, C. Yasunaga-Aoki, and S. Shimizu. 2011. Phylogenetic analysis of Metarhizium spp. isolated from soil in Japan. Appl. Entomol. Zool. 46:301-309. doi:10.1007/s13355-011-0045-y
Oulevey, C., F. Widmer, R. Kölliker, and J. Enkerli. 2009. An optimized microsatellite marker set for detection of Metarhizium anisopliae genotype diversity on field and regional scales. Mycol. Res. 113:1016-1024. doi:10.1016/j.mycres.2009.06.005
Peakall, R., and P.E. Smouse. 2006. GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol. Ecol. Notes 6:288-295. doi:10.1111/j.1471-8286.2005.01155.x
R-Core-Team. 2016. R: A language and environment for statistical computing. Foundation for Statistical Computing, Vienna, AUT.
Rehner, S.A., and E.P. Buckley. 2003. Isolation and characterization of microsatellite loci from the entomopathogenic fungus Beauveria bassiana (Ascomycota: Hypocreales). Mol. Ecol. Notes 3:409-411. doi:10.1046/j.1471-8286.2003.00464.x
Sánchez-Peña, S.R., J.S. Lara, and R.F. Medina. 2011. Occurrence of entomopathogenic fungi from agricultural and natural ecosystems in Saltillo, México, and their virulence towards thrips and whiteflies. J. Insect Sci. 11(1):1-10. doi:10.1673/031.011.0101
Schuelke, M. 2000. An economic method for the fluorescent labeling of PCR fragments. Nat. Biotechnol. 18:233-234. doi:10.1038/72708
Shete, S., H. Tiwari, and R.C. Elston. 2000. On estimating the heterozygosity and polymorphism information content value. Theor. Popul. Biol. 57:265-271. doi:10.1006/tpbi.2000.1452
St-Leger, R.J., B. May, L.L. Allee, D.C. Frank, R.C. Staples, and D.W. Roberts. 1992. Genetic differences in allozymes and in formation of infection structures among isolates of the entomopathogenic fungus Metarhizium anisopliae. J. Invertebr. Pathol. 60:89-101. doi:10.1016/0022-2011(92)90159-2
Toledo, A.V., M.E. Simurro, and P.A. Balatti. 2013. Morphological and molecular characterization of a fungus, Hirsutella sp., isolated from planthoppers and psocids in Argentina. J. Insect Sci. 13(18):18-11. doi:10.1673/031.013.1801
Wang, C., F.A. Shah, N. Patel, Z. Li, and T.M. Butt. 2003. Molecular investigation on strain genetic relatedness and population structure of Beauveria bassiana. Environ. Microbiol. 5:908-915. doi:10.1046/j.1462-2920.2003.00485.x
Zhang, S., X. Chen, F. Luan, L. He, S. Pu, and Z. Li. 2016. Genetic diversity and population structure of the Chinese fungus Metarhizium rileyi causing green muscardine in silkworm. J. Invertebr. Pathol. 140:16-24. doi:10.1016/j.jip.2016.08.005
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