Molecular characterization and virulence of the entomopathogenic fungus Beauveria bassiana from Ostrinia nubilalis (Lepidoptera: Pyralidae)

Year 2012, Volume: 2 Issue: 1, 23 - 30, 25.06.2012

İsmail Demir , Emrah Seçil Zihni Demirbağ , Ali Sevim

Abstract

The European corn borer (Ostrinia nubilalis, Lepidoptera: Pyralidae) is one of the most important pest of maize and other crops, such as potato, green pepper, and winter wheat almost all over the world. In this study, we collected larvae from corn fields invaded by O. nubilalis during 2008-2010 in the Eastern Black Sea Region of Turkey and investigated fungal pathogens of this pest. As result of this investigation, one fungal species was isolated from O. nubilalis and it was characterized according to morphological and molecular characteristics. Based on these characteristics, the isolate was identified as Beauveria bassiana. Additionally, insecticidal activity of the isolate was determined on O. nubilalis larvae by testing different concentrations of conidia (1 × 104, 1 × 105, 1 × 106, 1 × 107 and 1 × 108 conidia ml-1). It caused 80% mortality within 13 days after inoculation of 1 × 108 conidia ml-1. Mycosis was calculated as 70% and LC50 value was also calculated as 4.8 × 105 conidia ml-1. These results indicate that Beauveria bassiana Ost3 seems to be a good candidate as biocontrol agent against O. nubilalis.

Keywords

Beauveria bassiana, european corn borer, corn, LC50

Ostrinia nubilalis (Lepidoptera: Pyralidae)’den elde edilen entomopatojenik fungus Beauveria bassiana’nın moleküler karakterizasyonu ve virulansı

Abstract

Mısır kurdu (Ostrinia nubilalis, Lepidoptera: Pyralidae) hemen hemen bütün dünyada mısır, patates, yeşil biber ve kışlık buğday gibi pek çok ürünün en önemli zararlılarından birisidir. Bu çalışmada, 2008-2010 yılları arasında Doğu Karadeniz Bölgesi’nde O. nubilalis tarafından istila edilmiş mısır tarlalarından böcek örnekleri toplanmış ve entomopatojenik fungus yönünden araştırılmıştır. Bu araştırmanın sonucu olarak, O. nubilalis’ten bir fungal örnek izole edildi ve onun morfolojik ve moleküler verilere göre karakterizasyonu yapıldı. Bu karakterlere göre izolat Beauveria bassiana olarak tanımlandı. İlave olarak, bu izolatın O. nubilalis larvaları üzerindeki insektisidal aktivitesi farklı konsantrasyonlarda spor süspansiyonları (1 × 104, 1 × 105, 1 × 106, 1 × 107 and 1 × 108 spor/ml) kullanılarak belirlendi. Uygulama sonrasında, Ost3 1 × 108 spor/ml spor konsantrasyonun uygulanmasından sonra 13 gün içerisinde larvalar üzerinde %80 ölüm oranına neden oldu. Mikoz %70 ve LC50 değeri ise 4.8 × 105 spor/ml olarak hesaplandı. Bu sonuçlar, Beauveria bassiana Ost3’ün O. nubilalis’e karşı biyolojik mücadele etmeni olarak iyi bir aday olduğunu göstermektedir.

Keywords

Beauveria bassiana, european corn borer, corn, LC50

References

• Abbott, W. S., 1925. A method of computing the effectiveness of an insecticide. Journal of Economic Entomology, 18: 265-267.
• Anonymous, 2008. The Technical guide of agricultural control. Başak Publisher, Ankara, 262-265 pp.
• Bing, L. A. & L. C. Lewis, 1993. Occurence of the entomopathogen Beauveria bassiana (Bals.) Vuill. in different tillage regimes and in Zea mays L. and virulence towards Ostrinia nubilalis (Hbn.). Agriculture, Ecosystems and Environment, 45: 147-156.
• Cagan, L. & V. Uhlik, 1999. Pathogenicity of Beauveria bassiana strains isolated from Ostrinia nubilalis Hbn. (Lepidoptera: Pyrapidae) to original host and to ladybirds (Coleoptera: Coccinelidae). Plant Protection Science, 35: 108-112.
• Campbell, R. R., T. E. Anderson, M. Semel & D. W. Roberts, 1985. Management of the Colorado potato beetle using the entomogenous fungus Beauveria bassiana. American Potato Journal, 61: 29-37.
• Carruthers, R. I., Z. Feng, D. S. Robson & D. W. Roberts, 1985. In vivo temperature-dependent development of Beauveria bassiana (Deuteromycotina: Hyphomycetes) mycosis of the European corn borer, Ostrinia nubilalis (Lepidoptera: Pyralidae). Journal of Invertebrate Pathology, 46: 305-311.
• Chandler, D., G. Davidson, J. K. Pell, B. V. Ball, K. Shaw & K. D. Sunderland, 2000. Fungal biocontrol of acari. Biocontrol Science and Technology, 10: 357-384.
• Feng, Z., R. I. Carruthers, D. W. Roberts & D. S. Robson, 1985. Age-specific dose-mortality effects of Beauveria bassiana (Deuteromycotina: Hyphomycetes) on the European corn borer, Ostrinia nubilalis (Lepidoptera: Pyralidae). Journal of Invertebrate Pathology, 46: 259-264.
• Feng, Z, R. I. Carruthers & T. S. Larkin, 1988. A phenology model and field evaluation of Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) mycosis of the European corn borer, Ostrinia nubilalis (Hbn.) (Lepidoptera: Pyralidae). Canadian Entomology, 120: 133-144.
• Fuxa, J. R., 1987. Ecological considerations for the use of Entomopathogens in IPM. Annual Review of Entomology, 32: 225-51.
• Gaspers, C., 2009. The European corn borer (Ostrinia nubilalis, Hbn.), its susceptibility to the Bt-toxin Cry1F, its pheromone races and its gene flow in Europe in view of an Insect Resistance Management. PhD thesis, RWTH Aachen University, 113 p.
• Glare, T. R. & A. Inwood, 1998. Morphological and genetic characterization of Beauveria spp. from New Zealand. Mycological Research, 102: 250-256.
• Glare, T. R., 2004. Molecular characterization in the entomopathogenic fungal genus Beauveria. Laimburg Journal, 1: 286-298.
• Glare, T. R., S. D. Reay, T. L. Nelson & R. Moore, 2008. Beauveria caledonica is a naturally occurring pathogen of forest beetles. Mycological Research, 112: 352-360.
• Goettel, M. S., J. Eilenberg & T. Glare, 2005. “Entomopathogenic fungi and their role in regulation of insect populations, 361-405 ”. In: Comprehensive Molecular Insect Science (Eds. L.I. Gilbert, K. Iatrou & S.S. Gill), Amsterdam) 3300pp.
• Gökçe, A. & M. K. Er, 2005. Pathogenicity of Paecilomyces spp. to the glasshouse whitefly, Trialeurodes vaporariorum, with some observations on the fungal infection process. Turkish Journal of Agriculture and Forestry, 29: 331-339.
• Hall, T.A, 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium, 41: 95-98.
• Hoffmann, M. P., M. G. Wright, S. A. Pitcher & J. Gardner, 2002. Inoculative releases of Trichogramma ostriniae for suppression of Ostrinia nubilalis (European corn borer) in sweet corn: field biology and population dynamics. Biological Control, 25: 249-258.
• Humber, R., 1997. “Entomopathogenic Fungal Identification,153-185”. In: Manual of Techniques In Insect Pathology, (Eds. L.A. Lacey), San Diego, 409 pp.
• Inglis, G. D., M. S. Goettel, T. M. Butt & H. Strasser, 2001. “Use Of Hyphomycetous Fungi For Managing Insect Pests, 23-69 ”. In: Fungi As Biocontrol Agents (Eds. T. M. Butt, C. Jackson & N. Magan), Oxon, 416 pp.
• Kimura, M., 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16: 111-120.
• Leathers, T. D. & S. C. Gupta, 1993. Susceptibility of the eastern tent caterpillar (Malacosoma americanum) to the entomogenous fungus Beauveria bassiana. Journal of Invertebrate Pathology, 61: 217–219.
• Legaspi, J. C., T. J. Poprawski & B. C. Legaspi, 2000. Laboratory and field evaluation of Beauveria bassiana against sugarcane stalkborers (Lepidoptera: Pyralidae) in the lower Rio Grande Valley of Texas. Journal of Economic Entomology, 93: 54-9.
• Lewis, L. C., 1975. Natural regulation of crop pests in their indigenous ecosystems and in Iowa agrosystems: bioregulation of economic insect pests. Iowa State Journal of Research, 49: 435-445.
• Meyling, N. V. & J. Eilenberg, 2007. Ecology of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae in temperate agroecosystems: Potential for conservation biological control. Biological Control, 43: 145-155.
• Padmaja, V. & G. Kaur, 2001. Use of the fungus Beauveria bassiana (Bals.) Vuill (Moniliales: Deuteromycetes) for controlling termites. Current Science, 81: 25.
• Rehner, S. A. & E. Buckley, 2005. A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia, 97: 84-98.
• Sevim, A., I. Demir & Z. Demirbag, 2010a. Molecular Characterization and Virulence of Beauveria spp. from the Pine Processionary Moth, Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae). Mycopathologia, 170: 269–277.
• Sevim, A., I. Demir, M. Höfte, R. A. Humber & Z. Demirbag, 2010b. Isolation and characterization of entomopathogenic fungi from hazelnut-growing region of Turkey. BioControl, 55: 279-297.
• Shah, P. A., & J. K. Pell, 2003. Entomopathogenic fungi as biological control agents. Applied Microbiology and Biotechnology, 61: 413–423.
• St Leger, R. & D. W. Roberts, 1997. Engineering improved mycoinsecticides. Trends Biotechnology, 15: 83-85.
• Tafoya, F., M. Zuniga-Delgadillo, R. Alatorre, J. Cibrian-Tovar & D. Stanley, 2004. Pathogenicity of Beauveria Bassiana (Deuteromycota: Hyphomycetes) against the Cactus Weevil, Metamasius Spinolae (Coleoptera: Curculionidae) under laboratory conditions. Florida Entomology, 87: 4.
• Tamura, K., J. Dudley, M. Nei & S. Kumar, 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24: 1596-1599.
• Todorova, S. I., C. Cloutier, J. C. Cote & D. Coderre, 2002. Pathogenicity of six isolates of Beauveria bassiana (Balsamo) Vuillemin (Deuteromycotina, Hyphomycetes) to Perillus bioculatus (F) (Hem., Pentatomidae). Journal of Applied Entomology, 126: 182-185.
• Wagner, B. & L. C. Lewis, 2000. Colonization of corn, Zea mays, by the entomopathogenic fungus Beauveria bassiana. Applied and Environmental Microbiology, 66: 3468–3473.
• White, T. J., T. Bruns, S. Lee & J. Taylor, 1990. “Amplification And Direct Sequencing Of Fungal Ribosomal Rna Genes For Phylogenetics, 315-322”. In: PCR Protocols: A Guide to Methods and Applications (Eds: M. A. Innis, D. H. Gelfand, J. J. Sninsky & T. J. White), San Diego, 482 pp.
• Wraight, S. P., M. E. Ramos, P. B. Avery, S. T. Jaronski & J. D. Vandenberg, 2010. Comparative virulence of Beauveria bassiana isolates against lepidopteran pests of vegetable crops. Journal of Invertebrate Pathology, 103: 186–199.
• Zurek, L. & B. A. Keddie, 2000. Beauveria bassiana (Balsamo) Vuillemin - a promising microbial control agent of the satin moth (Lepidoptera: Lymantriidae). Biocontrol Science and Technology, 10: 641-644.