Occurrence of Entomopathogenic Fungi from Natural Ecosystems in Phetchabun, Thailand and Their Virulence Against Brown Planthopper

Main Article Content

Araya Bunsak
Wipa Homhaul
Weerathep Pongprasert
Krissana Ruangrit
Suphannika Intanon*

Abstract

Brown planthopper (BPH), Nilaparvata lugens Stål, is a serious insect pest of rice. The invasion of BPH causes heavy losses in rice production, both quality and quantity. The objective of this research was to search for Metarhizium spp., a genus of entomopathogenic fungi in the Clavicipitaceae family, from forest soil in three districts of Phetchabun Province, Thailand: Khao Kho, Lom Kao and Nam Nao to control BPH. Metarhizium spp. fungi were isolated by soil dilution plate technique and determined their efficacy using conidia suspension at the concentration of 1X108 conidia/mL on BPH at the 2-3th instar nymph fed on seedlings of susceptible rice cultivars, Taichung Native 1 (TN1). The most effective Metarhizium sp. isolate was identified using the ITS region of 18S rDNA sequencing, Basic Local Alignment Search Tool (BLAST) and evolutionary history. The result showed that 126 isolates of Metarhizium spp. were found and coded as PB-01 to PB-126. All isolates infected BPH nymph with 26.7 to 100% mortality within 6 days after contacting conidia suspension. The isolate of PB-75 showed the highest efficacy (100% BPH mortality) with the lethal times of 50% mortality (LT50) within 2.9 days. The species identification showed that the DNA sequence of Metarhizium sp. isolate, PB-75, was 98.6% similar to M. anisopliae Genbank ID JQ889704.1. Evolutionary history based on phylogenetic analysis using neighbor-joining and maximum parsimony methods confirmed that PB-75 formed the same phylogenetic clade with M. anisopliae Genbank ID JQ889704.1 and M. anisopliae var. anisopliae.


Keywords: biological control; brown planthopper; Metarhizium spp.; phylogenetic analysis; rice


*Corresponding author: E-mail: [email protected]

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Original Research Articles

References

Dyck, V.A., and Thomas, B., 1979. The brown planthopper problem. In: International Rice Research Institute, ed. Brown Planthopper: Threat to Rice Production in Asia. Los Baños: International Rice Research Institute, pp. 3-20.

Ruay-aree, S., 2001. Learning Management of Rice Pest by Integrated Method. Bangkok: The Agricultural Cooperative Federation of Thailand Limited. (in Thai)

Hu, G., Cheng, X.N., Guo-Jun, Q., Wang, F.Y., Lu,F., Zhang, X.X. and Zhai, B.P., 2010. Rice planting systems, global warming and outbreaks of Nilaparvata lugens (Stål). Bulletin of Entomological Research, 101, 187-199.

Kari, I. and Danar, D., 2021. A review on brown planthopper (Nilaparvata lugens Stål), a major pest of rice in asia and pacific. Asian Journal of Research in Crop Science, 6, 7-19.

Sōgawa, K., 1982. The rice brown planthopper: feeding physiology and host plant interactions. Annual review of entomology, 27(1), 49-73.

Seo, B.Y., Kwon, Y.-H., Jung, J.K. and Kim, G.-H., 2009. Electrical penetration graphic waveforms in relation to the actual positions of the stylet tips of Nilaparvata lugens in rice tissue. Journal of Asia-Pacific Entomology, 12(2), 89-95.

Rubia-Sachez, E., Suzuki, Y., Arimura, K., Miyamoto, K., Matsumura, M. and Watanabe, T., 2003. Comparing Nilaparvata lugens (Stal) and Sogatella furcifera (Horvath) (Homoptera: Delphacidae) feeding effects on rice plant growth processes at the vegetative stage. Crop Protection, 22, 967-974.

Xu, H.X., He, X.C., Zheng, X.S., Yang, Y.J. and Lu, Z.X., 2014. Influence of rice black streaked dwarf virus on the ecological fitness of non‐vector planthopper Nilaparvata lugens (Hemiptera: Delphacidae). Insect Science, 21(4), 507-514.

Baehaki, S., Iswanto, E.H., Munawar, D., Song, Y.-H., Choi, I. and Park, H.-H., 2017. Abilities of brown planthopper immigrant transmits rice ragged stunt virus on rice of some district of Java-Indonesia. Scholars Journal of Agriculture and Veterinary Sciences, 4(8), 300-310.

Helina, S., Sulandari, S., Hartono, S., and Trisyono, Y., 2019. Detection and analysis of protein profile on rice infected by stunting virus with different severity on ciherang and situ bagendit varieties. Journal Perlindungan Tanaman Indonesia, 23, DOI: 10.22146/jpti.36549.

Cheng, X., Wu, J. and Ma, F., 2003. Brown Planthopper: Occurrence and Control. Beijing: China Agricultural Press.

Backus, E.A., Serrano, M.S. and Ranger, C.M., 2005. Mechanisms of hopperburn: an overview of insect taxonomy, behavior, and physiology. Annual Review of Entomology, 50, 125-151.

Cheng, J., 2015. Rice planthoppers in the past half century in China. In: K.L. Heong, J. Cheng and M.M. Escalada, eds. Rice Planthoppers: Ecology, Management, Socio Economics and Policy, Amsterdam: Springer, pp. 1-32.

Ko, K., Liu, Y., Hou, M., Babendreier, D., Zhang, F. and Song, K., 2015. Toxicity of insecticides targeting rice planthoppers to adult and immature stages of Trichogramma chilonis (Hymenoptera: Trichogrammatidae). Journal of Economic Entomology, 108(1), 69-76.

Heong, K.L., 2009. Are planthopper problems caused by a breakdown in ecosystem services? In: K. Heong and B.L. Hardy, eds. Planthoppers: New Threats to the Sustainability of Intensive Rice Production Systems in Asia. Los Baños: International Rice Research Institute, pp. 221-232.

Nagata, T., 2002. Monitoring on insecticide resistance of the brown planthopper and the white backed planthopper in Asia. Journal of Asia-Pacific Entomology, 5(1), 103-111.

Matsumura, M., Takeuchi, H., Satoh, M., Sanada-Morimura, S., Otuka, A., Watanabe, T. and Thanh, D.V., 2009. Current status of insecticide resistance in rice planthoppers in Asia. In: K.L. Heong and B. Hardy, eds. Planthoppers: New Threats to the Sustainability of Intensive Rice Production Systems in Asia. Los Baños: International Rice Research Institute, pp. 233-243.

Matsumura, M., and Sanada-Morimura, S., 2010. Recent Status of Insecticide Resistance in Asian Rice Planthoppers. Japan Agricultural Research Quarterly, 44(3), 225-230.

Ali, M.P., Huang, D., Nachman, G., Ahmed, N., Begum, M.A. and Rabbi, M.F., 2014. Will climate change affect outbreak patterns of planthoppers in Bangladesh? PLoS ONE, 9(3), DOI: 10.1371/journal.pone.0091678.

Jin, S.F., Feng, M.G. and Chen, J.Q., 2008. Selection of global Metarhizium isolates for the control of the rice pest Nilaparvata lugens (Homoptera: Delphacidae). Pest Management Science, 64(10), 1008-1014. DOI: 10.1002/ps.1597.

Litwin, A., Nowak, M. and Różalska, S., 2020. Entomopathogenic fungi: unconventional applications. Reviews in Environmental Science and Biotechnology, 19(1), 23-42.

Zimmermann, G., 2007. Review on safety of the entomopathogenic fungus Metarhizium anisopliae. Biocontrol Science and Technology, 17(9), 879-920.

Shahid, A.A., Rao, Q.A., Bakhsh, A. and Husnain, T., 2012. Entomopathogenic fungi as biological controllers: new insights into their virulence and pathogenicity. Archives of Biological Sciences, 64(1), 21-42.

Gibson, D.M., Donzelli, B.G., Krasnoff, S.B. and Keyhani, N.O., 2014. Discovering the secondary metabolite potential encoded within entomopathogenic fungi. Natural Product Reports, 31(10), 1287-1305.

Maketon, C., Buapha, S., Rungratanaubon, T. and Maketon, M., 2015. Laboratory and field evaluations of Beauveria bassiana (Bals.-Criv.) Vuill. and Metarhizium robertsii (J.F. Bisch, Rehner and Humber) against the brown plant hopper, Nilaparvata lugens Stal and its natural enemies in paddy fields in Thailand. Egyptian Journal of Pest Control, 25, 97-105.

Gul, H., Saeed, S. and Khan, F.Z.A., 2014. Entomopathogenic fungi as effective insect pest management tactic: a review. Applied Sciences and Business Economics, 1(1), 10-18.

Roy, H.E. and Pell, J.K., 2000. Interactions between entomopathogenic fungi and other natural enemies: implications for biological control. Biocontrol Science and Technology, 10(6), 737-752.

Reddy, K.R.K, PraveenKumar, D. and Reddy, K.R.N., 2013. Entomopathogenic fungi: a potential bioinsecticide. Kavaka, 41, 23-32.

Chinniah, C.C.H., Ravikumar, A., Kalyanasundaram, M. and Parthiban, P., 2016. Management of sucking pests, by integration of organic sources of amendments and foliar application of entomopathogenic fungi on chilli. Journal of Biopesticides, 9(1), 34-40.

Hywel-Jones, N.L. and Gillespie, A.T., 1990. Effect of temperature on spore germination in Metarhizium anisopliae and Beauveria bassiana. Mycological Research, 94(3), 389-392.

Hsia, I.C.C., Islam, M.T., Yusof, I., How, T.Y., and Omar, D., 2014. Evaluation of conidial viability of entomopathogenic fungi as influenced by temperature and additive. International Journal of Agriculture and Biology, 16(1), 146-152.

Acheampong, M.A., Hill, M.P., Moore, S.D. and Coombes, C.A., 2020. UV sensitivity of Beauveria bassiana and Metarhizium anisopliae isolates under investigation as potential biological control agents in South African citrus orchards. Fungal Biology, 124(5), 304-310, DOI: 10.1016/j.funbio.2019.08.009.

Sharma, L., Oliveira, I., Gonçalves, F., Raimundo, F., Singh, R.K., Torres, L. and Marques, G., 2021. Effect of soil chemical properties on the occurrence and distribution of entomopathogenic fungi in Portuguese grapevine fields. Pathogens, 10(2), DOI: 10.3390/pathogens10020137.

Chen, W.-H., Han, Y.-F., Liang, J.-D. and Liang, Z.-Q., 2019. Morphological and phylogenetic characterization of novel Metarhizium species in Guizhou, China. Phytotaxa, 419(2), 189-196.

Masoudi, A., Wang, M., Zhang, X., Wang, C., Qiu, Z., Wang, W., Wang, H. and Liu, J., 2020. Meta-analysis and evaluation by insect-mediated baiting reveal different patterns of hypocrealean entomopathogenic fungi in the soils from two regions of China. Frontiers in Microbiology, 11, DOI: 10.3389/fmicb.2020.01133.

Inglis, G.D., Enkerli, J. and Goettel, M.S., 2012. Laboratory techniques used for entomopathogenic fungi: Hypocreales. In: L.A. Lacey, ed. Manual of Techniques in Invertebrate Pathology. San Diego: Academic Press, pp. 189-253.

Fernandes, É.K.K., Keyser, C.A., Rangel, D.E.N., Foster, R.N. and Roberts, D.W., 2010. CTC medium: A novel dodine-free selective medium for isolating entomopathogenic fungi, especially Metarhizium acridum, from soil. Biological Control, 54(3), 197-205.

Goettel, M.S. and Inglis, D.G. 1997. Fungi: Hyphomycetes. In: L. Lacey, ed. Manual of Techniques in Insect Pathology. London: Acadamic press, pp. 213-249.

Gilman, J.C., 1957. A Manual of Soil Fungi. revised 2nd ed. Calcutta: India Oxford and IBH Publishing Company.

Humber, R.A., 2005. Entomopathogenic Fungal Identification. [online] Available at: https://www.ars.usda.gov/ARSUserFiles/80620520/apswkshoprev.pdf.

Sevim, A., Demir, I., Höfte, M., Humber, R.A. and Demirbag, Z., 2010. Isolation and characterization of entomopathogenic fungi from hazelnut-growing region of Turkey. Biocontrol, 55(2), 279-297.

Abbott, W.S., 1925. A method of computing the effectiveness of an insecticide. Journal of Economic Entomology,18, 265-267.

Finney, D.J., 1971. Probit Analysis. 3rd ed. Cambridge: Cambridge University Press.

White, T.J., Bruns, T., Lee, S. and Taylor, J., 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: M.A. Innis, D.H. Gelfand, J.J. Shinsky and T.J. White, eds. PCR Protocols: A Guide to Methods and Applications. New York: Academic Press, Inc., pp. 315-322.

Hall, T.A., 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98.

Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F. and Higgins, D.G., 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25(24), 4876-4882.

Hall, B.G., 2013. Building phylogenetic trees from molecular data with MEGA. Molecular Biology and Evolution, 30(5), 1229-1235, DOI: 10.1093/molbev/mst012.

Tamura, K., Stecher, G. and Kumar, S., 2021. MEGA11: molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution, 38(7), 3022-3027.

Butt, T.M., Wang, C., Shah, F.A. and Hall, R., 2006. Degeneration of entomogenous fungi. In J. Eilenberg and H.M.T. Hockkanen, eds. An Ecological and Societal Approach to Biological Control. Dordrecht: Springer, pp. 213-226.

Keller, S. and Zimmerman, G., 1989. Mycopathogens of soil insects. In: N. Wilding, N.M. Collins, P.M. Hammond and J.F. Webber, eds. Insect-Fungus Interactions. London: Academic Press, pp. 240-270.

Aung, O.M., Oytong, K. and Hyde, K.D., 2008. Diversity of entomopathogenic fungi in rainforests of Chiang Mai Province, Thailand. Fungal Diversity, 30, 15-22.

Miętkiewski, R., Żurek, M., Tkaczuk, C. and Bałazy, S., 1991. Occurrence of entomopathogenic fungi in arable soil, forest soil and litter. Roczniki Nauk Rolniczych, 21, 61-68.

Sánchez-Peña, S.R., Lara, J.S.-J. and Medina, R.F., 2011. Occurrence of entomopathogenic fungi from agricultural and natural ecosystems in Saltillo, México, and their virulence towards thrips and whiteflies. Journal of Insect Science, 11(1), DOI: 10.1673/031.011.0101.

Ayele, B.A., Muleta, D., Venegas, J. and Assefa, F., 2020. Morphological, molecular, and pathogenicity characteristics of the native isolates of Metarhizium anisopliae against the tomato leafminer, Tuta absoluta (Meyrick 1917) (Lepidoptera: Gelechiidae) in Ethiopia. Egyptian Journal of Biological Pest Control, 30(1), DOI: 10.1186/s41938-020-00261-w.

Tangthirasunun, N., Poeaim, S., Soytong, K., Sommartya, P. and Popoonsak, S., 2010. Variation in morphology and ribosomal DNA among isolates of Metarhizium anisopliae from Thailand. Journal of Agricultural Technology, 6(2), 317-329.

Ma, W.S., Qiao, H.L., Nong, X.Q., Chen, J., Yu, J., Qin, R.M., Xu, C.Q., Liu, S., Li, X.M., and Cheng, H.Z., 2013. Screening for virulence strains of Metarhizium against Dorysthenes hydropicus Pascoes. China Journal of Chinese Materia Medica, 38, 3438-3441.

Aw, K.M.S. and Hue, S.M., 2017. Mode of infection of Metarhizium spp. fungus and their potential as biological control agents. Journal of Fungi (Basel), 3(2), DOI: 10.3390/jof3020030.

Tang, J., Liu, X., Ding, Y., Jiang, W. and Xie, J., 2019. Evaluation of Metarhizium anisopliae for rice planthopper control and its synergy with selected insecticides. Crop Protection, 121, 132-138.

Bunsak, A., Pongprasert, W., Buranapanichpan, S. and Kulsarin, J., 2015. Screening on potential Metarhizium anisopliae (Metschnikoff) Sorokin forcontrolling brown planthopper in paddy field. Journal of Agriculture, 31(3), 291-299. (In Thai)

Mohan, C., Sridhar, R.P., and Nakkeeran, S., 2016. Studies on efficacy of entomopathogenic fungi Metarhizium anisopliae (Metchnikoff) Sorokin against Nilaparvata lugens (Stål). International Journal of Agricultural Science Research, 6(6), 227-234.

Samuels, K.D.Z., Heale, J.B. and Llewellyn, M., 1989. Characteristics relating to the pathogenicity of Metarhizium anisopliae toward Nilaparvata lugens. Journal of Invertebrate Pathology, 53(1), 25-31, DOI: 10.1016/0022-2011(89)90070-0.

Bich, G.A., Castrillo, M.L., Kramer, F.L., Villalba, L.L. and Zapata, P.D., 2021. Morphological and molecular identification of entomopathogenic fungi from agricultural and forestry crops. Floresta e Ambiente, 28(2), DOI: 10.1590/2179-8087-floram-2018-0086.

Henry, T., Iwen, P.C. and Hinrichs, S.H., 2000. Identification of Aspergillus species using internal transcribed spacer regions 1 and 2. Journal of Clinical Microbiology, 38(4), 1510-1515.

Saitou, N. and Nei, M., 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4(4), 406-425.

Tamura, K., Nei, M. and Kumar, S., 2004. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proceedings of the National Academy of Sciences, 101(30), 11030-11035.

Felsenstein, J., 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39(4), 783-791.

Nei, M. and Kumar, S., 2000. Molecular Evolution and Phylogenetics. Oxford: Oxford University Press.

Xu, Y.L., Bao, S.X., Huang, H.Q., Liu, M. and Zhu, J., 2012. Diversity and Distribution of Culturable Fungi in Mangrove Soil in Hainan. [online] Available at: https://www.ncbi.nlm.nih.gov/nucleotide/JQ889704.1report=genbank&log$=nucltop&blast_rank=1&RID=U83FU99601N.

Kershaw, M.J., Moorhouse, E.R., Bateman, R., Reynolds, S.E. and Charnley, A.K., 1999. The role of destruxins in the pathogenicity of Metarhizium anisopliae for three species of insect. The Journal of Invertebrate Pathology, 74(3), 213-223.

Destéfano, R.H.R., Destefano, S.A.L. and Messias, C.L., 2004. Detection of Metarhizium anisopliae var. anisopliae within infected sugarcane borer Diatraea saccharalis (Lepidoptera, Pyralidae) using specific primers. Genetics and Molecular Biology, 27, DOI: 10.1590/S1415-47572004000200020.