Effectiveness of the entomopathogenic fungal strain Beauveria bassiana against male and female poplar/willow weevil (Cryptorhynchus lapathi) adults

Abstract

Cryptorhynchus lapathi L., a globally distributed wood-boring pest, significantly threatens poplar (Populus spp.) and willow (Salix spp.) trees. Entomopathogenic fungi (EPFs), particularly Beauveria bassiana (Bals.-Criv.) Vuilla, play a crucial role in integrated pest management because of their broad-spectrum insecticidal properties. This study clarifies the pathogenic mechanisms of B. bassiana CFCC81428 treatment and its oil-based formulation against male and female C. lapathi adults, alongside their impacts on cumulative mortality, enzyme activity and nutrient metabolism. The results indicated that the oil-based formulation significantly enhanced B. bassiana pathogenicity, achieving cumulative mortality rates of 91.7% and 83.8% for male and female adults, respectively, with LT₅₀ values of 7.9 and 8.7 d, markedly outperforming the B. bassiana treatment. Scanning electron microscopy revealed that the oil-based formulation improved spore adhesion to the insects’ cuticle, accelerated spore germination and hyphal growth, and significantly improved cuticular penetration efficiency. Within the host, the activities of detoxification enzymes (GST and CarE) were upregulated, whereas the activities of catalase and peroxidase enzymes were suppressed. Superoxide dismutase activity remained elevated throughout most of the observation time points. Polyphenol oxidase activity significantly increased between 24 and 120 h postinfection, indicating its critical role in preventing B. bassiana invasion. The infection also triggered substantial shifts in the host’s nutrient metabolism, with time-dependent changes observed in carbohydrates, free fatty acids, and soluble proteins. The oil-based formulation exacerbated the depletion of these nutrients, ultimately leading to metabolic collapse. This study indicates that the oil-based formulation optimizes spore germination conditions and accelerates infection, thereby significantly increasing B. bassiana pathogenicity in C. lapathi males, whereas female adults exhibited stronger physiological and metabolic responses, providing new insights for the application of B. bassiana in the biological control of pests.

Keywords

Biological control / Beauveria bassiana / Oil-based formulation / Cryptorhynchus lapathi / Gender-specific responses

References

[1] Acheampong MA, Hill MP, Moore SD, Coombes CA. UV sensitivity of Beauveria bassiana and Metarhizium anisopliae isolates under investigation as potential biological control agents in South African Citrus orchards. Fungal Biol, 2020, 124(5): 304-310

[2] An Y, Liu Y, Liu YJ, Lu MZ, Kang XH, Mansfield SD, Zeng W, Zhang J. Opportunities and barriers for biofuel and bioenergy production from poplar. GCB Bioenergy, 2021, 13(6): 905-913

[3] Apirajkamol N, Hogarty TM, Mainali B, Taylor PW, Walsh TK, Tay WT. Virulence of Beauveria sp. and Metarhizium sp. fungi towards fall armyworm (Spodoptera frugiperda). Arch Microbiol, 2023, 205(10 328

[4] Aw KMS, Hue SM. Mode of infection of Metarhizium spp. fungus and their potential as biological control agents. J Fungi, 2017, 3(2): 30

[5] Bateman RP, Carey M, Moore D, Prior C. The enhanced infectivity of Metarhizium flavoviride in oil formulations to desert locusts at low humidities. Ann Appl Biol, 1993, 122(1145-152

[6] Bhadani RV, Gajera HP, Hirpara DG, Kachhadiya HJ. Characterization and bio-efficacy of entomopathogenic Beauveria associated with cuticle-degrading enzymes to restrain sucking pest Bemisia tabaci. Parasitol Res, 2022, 121(7): 2019-2031

[7] Bhattacharyya PN, Sarmah SR, Roy S, Sarma B, Nath BC, Bhattacharyya LH. Perspectives of Beauveria bassiana, an entomopathogenic fungus for the control of insect-pests in tea [Camellia sinensis (L.) O. Kuntze]: opportunities and challenges. Int J Trop Insect Sci, 2023, 43(1): 1-19

[8] Broberg CL, Borden JH. Hybrid poplar clones with Populus maximowiczii parentage demonstrate postoviposition antibiosis to Cryptorhynchus lapathi (Coleoptera: Curculionidae). J Econ Entomol, 2005, 98(6): 2254-2259

[9] Broberg CL, Borden JH, Humble LM. Distribution and abundance of Cryptorhynchus lapathi on Salix spp. in British Columbia. Can J for Res, 2002, 32(3): 561-568

[10] Broberg CL, Borden JH, Gries R. Olfactory and feeding preferences of Cryptorhynchus lapathi L. (Coleoptera: Curculionidae) among hybrid clones and natural poplars. En, 2005, 34(6): 1606-1613

[11] Broberg CL, Borden JH, Gries R. Olfactory and feeding preferences of Cryptorhynchus lapathi among hosts and nonhosts. Can Entomol, 2006, 138(3): 357-366

[12] Broberg CL, Howard Inkster JA, Borden JH. Phenological and chemical differences among hybrid poplar clones (Salicaceae) varying in resistance to Cryptorhynchus lapathi (L.) (Coleoptera: Curculionidae). Biochem Syst Ecol, 2010, 38(1): 29-48

[13] Cao QJ, Yu J, Ran YL, Chi DF. Effects of plant volatiles on electrophysiological and behavioral responses of Cryptorrhynchus lapathi. Entomol Exp Appl, 2015, 156(2): 105-116

[14] Cao GC, Jia M, Zhao X, Wang L, Tu XB, Wang GJ, Nong XQ, Zhang ZH. Different effects of Metarhizium anisopliae strains IMI330189 and IBC200614 on enzymes activities and hemocytes of Locusta migratoria L. PLoS ONE, 2016, 11(5 e0155257

[15] Chang CC, Rahmawaty A, Chang ZW. Molecular and immunological responses of the giant freshwater prawn, Macrobrachium rosenbergii, to the organophosphorus insecticide, trichlorfon. Aquat Toxicol, 2013, 130: 18-26

[16] Ding SY, Li HY, Li XF, Zhang ZY. Effects of two kinds of transgenic poplar on protective enzymes system in the midgut of larvae of American white moth. J for Res, 2001, 12(2): 119-122

[17] Ding JN, Zhang HH, Chi DF. Effects of a pathogenic Beauveria bassiana (Hypocreales: Cordycipitaceae) strain on detoxifying and protective enzyme activities in Xylotrechus rusticus (Coleoptera: Cerambycidae) larvae. Fla Entomol, 2015, 98(4): 1148-1156

[18] Dubovskiy IM, Slyamova ND, Kryukov VY, Yaroslavtseva ON, Levchenko MV, Belgibaeva AB, Adilkhankyzy A, Glupov VV. The activity of nonspecific esterases and glutathione-S-transferase in Locusta migratoria larvae infected with the fungus Metarhizium anisopliae (Ascomycota, Hypocreales). Entomol Rev, 2012, 92(1): 27-31

[19] Feng MG, Pu XY, Ying SH, Wang YG. Field trials of an oil-based emulsifiable formulation of Beauveria bassiana conidia and low application rates of imidacloprid for control of false-eye leafhopper Empoasca vitis on tea in Southern China. Crop Prot, 2004, 23(6): 489-496

[20] Greenfield BPJ, Lord AM, Dudley E, Butt TM. Conidia of the insect pathogenic fungus, Metarhizium anisopliae, fail to adhere to mosquito larval cuticle. R Soc Open Sci, 2014, 1(2 140193

[21] Guo HR, Jia NY, Chen HW, Xie D, Chi DF. Preliminary analysis of transcriptome response of Dioryctria sylvestrella (Lepidoptera: Pyralidae) larvae infected with Beauveria bassiana under short-term starvation. InSects, 2023, 14(5): 409

[22] Gupta A, Nair S. Dynamics of insect-microbiome interaction influence host and microbial symbiont. Front Microbiol, 2020, 11: 1357

[23] Gürlek S, Sevim A, Sezgin FM, Sevim E. Isolation and characterization of Beauveria and Metarhizium spp. from walnut fields and their pathogenicity against the codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae). Egypt J Biol Pest Contr, 2018, 28(150

[24] Gutiérrez Y, Alarcón KA, Ortiz C, Santos-Holguín JM, García-Riaño JL, Mejía C, Amaya CV, Uribe-Gutiérrez L. Isolation and characterization of a native strain of the entomopathogenic fungus Beauveria bassiana for the control of the palm weevil Dynamis borassi (Coleoptera: Curculionidae) in the neotropics. World J Microbiol Biotechnol, 2024, 40(9260

[25] Hannon ER, Kittelson NT, Eaton JA, Brown JJ. Screening hybrid poplar clones for susceptibility to Cryptorhynchus lapathi (Coleoptera: curculionidae). J Econ Entomol, 2008, 101(1199-205

[26] Hassuba MMM, Gad HA, Atta AAM, Abdelgaleil SAM. Efficacy of entomopathogenic fungi for the management of Trogoderma granarium Everts on wheat grains. Int J Trop Insect Sci, 2024, 44(31367-1374

[27] Islam SMN, Chowdhury MZH, Mim MF, Momtaz MB, Islam T. Biocontrol potential of native isolates of Beauveria bassiana against cotton leafworm Spodoptera litura (Fabricius). Sci Rep, 2023, 13(1): 8331

[28] Ismail SM. Effect of sublethal doses of some insecticides and their role on detoxication enzymes and protein-content of Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae). Bull Natl Res Cent, 2020, 44(1 35

[29] Jia HR, Camara I, Zhang Z, Gao YR, Yang XQ, Sangbaramou R, Zhen CA, Shi WP, Tan SQ. Effect of ultraviolet radiation on Beauveria bassiana virulence and development of protective formulations. Arch Microbiol, 2023, 205(4112

[30] Jiang WJ, Peng YF, Ye JY, Wen YY, Liu GX, Xie JQ. Effects of the entomopathogenic fungus Metarhizium anisopliae on the mortality and immune response of Locusta migratoria. InSects, 2019, 11(136

[31] Kaiser D, Handschin S, Rohr RP, Bacher S, Grabenweger G. Co-formulation of Beauveria bassiana with natural substances to control pollen beetles–synergy between fungal spores and colza oil. Biol Control, 2020, 140 104106

[32] Khamis WM, El-Sabrout AM, Shahin R, Abdel-Rahim EF. Field efficacy, sub-lethal, and biochemical effects of certain biorational insecticides against the new intruder, Spodoptera frugiperda in Bani-Suef, upper Egypt. Neotrop Entomol, 2023, 52(5963-973

[33] Khun KK, Ash GJ, Stevens MM, Huwer RK, Wilson BAL. Transmission of Metarhizium anisopliae and Beauveria bassiana to adults of Kuschelorhynchus macadamiae (Coleoptera: Curculionidae) from infected adults and conidiated cadavers. Sci Rep, 2021, 11(1): 2188

[34] Kraaijeveld AR, Barker CL, Godfray HCJ. Stage-specific sex differences in Drosophila immunity to parasites and pathogens. Evol Ecol, 2008, 22(2217-228

[35] Lebrun M, Miard F, Nandillon R, Léger JC, Hattab-Hambli N, Scippa GS, Bourgerie S, Morabito D. Assisted phytostabilization of a multicontaminated mine technosol using biochar amendment: early stage evaluation of biochar feedstock and particle size effects on As and Pb accumulation of two Salicaceae species (Salix viminalis and Populus euramericana). Chemosphere, 2018, 194: 316-326

[36] Lee KP, Cory JS, Wilson K, Raubenheimer D, Simpson SJ. Flexible diet choice offsets protein costs of pathogen resistance in a caterpillar. Proc Biol Sci, 2006, 273(1588): 823-829

[37] Leemon DM, Jonsson NN. Laboratory studies on Australian isolates of Metarhizium anisopliae as a biopesticide for the cattle tick Boophilus microplus. J Invertebr Pathol, 2008, 97(1): 40-49

[38] Lei JH, Sun TF, Feng MG, Ying SH. Functional insights of three RING-finger peroxins in the life cycle of the insect pathogenic fungus Beauveria bassiana. Curr Genet, 2023, 69(4–6): 267-276

[39] Li HP, Dz H, Tang XG. Changes in the contents of proteins and amino acids of Apriona germari larvae infected by Beauveria bassiana. Scientia Silvae Inicae, 2012, 48(5): 159-163

[40] Li XY, Liu QZ, Lewis EE, Tarasco E. Activity changes of antioxidant and detoxifying enzymes in Tenebrio molitor (Coleoptera: Tenebrionidae) larvae infected by the entomopathogenic nematode Heterorhabditis beicherriana (Rhabditida: Heterorhabditidae). Parasitol Res, 2016, 115(12): 4485-4494

[41] Li ZQ, Yu H, Huang GH. Changes in lipid, protein and carbohydrate metabolism in Spodoptera exigua larvae associated with infection by Heliothis virescens ascovirus 3h. J Invertebr Pathol, 2018, 155: 55-63

[42] Litwin A, Nowak M, Różalska S. Entomopathogenic fungi: unconventional applications. Rev Environ Sci Bio/technol, 2020, 19(1): 23-42

[43] Lopes RB, Faria M. Influence of two formulation types and moisture levels on the storage stability and insecticidal activity of Beauveria bassiana. Biocontrol Sci Technol, 2019, 29(5): 437-450

[44] Mann AJ, Davis TS. Plant secondary metabolites and low temperature are the major limiting factors for Beauveria bassiana (Bals.-Criv.) Vuill. (Ascomycota: Hypocreales) growth and virulence in a bark beetle system. Biol Contr, 2020, 141: 104130

[45] Niu F, Xing Y, Jia NY, Ding K, Xie D, Chen HW, Chi DF. Effectiveness of entomopathogenic fungal strains against poplar/willow weevil (Cryptorhynchus lapathi L.) larvae. J Forestry Res, 2022, 33(5): 1691-1702

[46] Pachoute J, dos Santos GR, de Souza DJ. Antagonistic effects of Beauveria bassiana on seed-borne fungi of cowpea (Vigna unguiculata). Biologia, 2024, 79(51487-1495

[47] Paschapur AU, Subbanna ARNS, Singh AK, Jeevan B, Stanley J, Rajashekara H, Mishra KK, Koti PS, Kant L, Pattanayak A. Alternaria alternata strain VLH1: a potential entomopathogenic fungus native to North Western Indian Himalayas. Egypt J Biol Pest Contr, 2022, 32(1): 138

[48] Pavlidi N, Vontas J, Van Leeuwen T. The role of glutathione S-transferases (GSTs) in insecticide resistance in crop pests and disease vectors. Curr Opin Insect Sci, 2018, 27: 97-102

[49] Qu S, Wang SB. Interaction of entomopathogenic fungi with the host immune system. Dev Comp Immunol, 2018, 83: 96-103

[50] Ranesi M, Vitale S, Staropoli A, Di Lelio I, Izzo LG, De Luca MG, Becchimanzi A, Pennacchio F, Lorito M, Woo SL, Vinale F, Turrà D. Field isolates of Beauveria bassiana exhibit biological heterogeneity in multitrophic interactions of agricultural importance. Microbiol Res, 2024, 286 127819

[51] Shahbaz M, Palaniveloo K, Tan YS, Palasuberniam P, Ilyas N, Wiart C, Seelan JSS. Entomopathogenic fungi in crops protection with an emphasis on bioactive metabolites and biological activities. World J Microbiol Biotechnol, 2024, 40(7217

[52] Siva-Jothy MT, Moret Y, Rolff J. Simpson SJ. Insect immunity: an evolutionary ecology perspective. Advances in insect physiology, 2005, Sydney, The University of Sydney1-48 32

[53] Smith BD, Stott KG. The life history and behaviour of the willow weevil Cryptorrhynchus lapathi L. Ann Appl Biol, 1964, 54(1): 141-151

[54] Söderhäll K, Cerenius L. Role of the prophenoloxidase-activating system in invertebrate immunity. Curr Opin Immunol, 1998, 10(1): 23-28

[55] Stenberg JA, Sundh I, Becher PG, Björkman C, Dubey M, Egan PA, Friberg H, Gil JF, Jensen DF, Jonsson M, Karlsson M, Khalil S, Ninkovic V, Rehermann G, Vetukuri RR, Viketoft M. When is it biological control? A framework of definitions, mechanisms, and classifications. J Pest Sci, 2021, 94(3): 665-676

[56] Sun R, Hong BK, Reichelt M, Luck K, Mai DT, Jiang XC, Gershenzon J, Vassão DG. Metabolism of plant-derived toxins from its insect host increases the success of the entomopathogenic fungus Beauveria bassiana. ISME J, 2023, 17(10): 1693-1704

[57] Toledo AV, de Remes Lenicov AM, López Lastra CC. Histopathology caused by the entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae, in the adult planthopper, Peregrinus maidis, a maize virus vector. J Insect Sci, 2010, 10: 35

[58] Tseng MN, Chung CL, Tzean SS. Mechanisms relevant to the enhanced virulence of a dihydroxynaphthalene-melanin metabolically engineered entomopathogen. PLoS ONE, 2014, 9(3 e90473

[59] Vertyporokh L, Hułas-Stasiak M, Wojda I. Host-pathogen interaction after infection of Galleria mellonella with the filamentous fungus Beauveria bassiana. Insect Sci, 2020, 27(5): 1079-1089

[60] Wang YC, Chi DF. Screening of high-virulent entomopathogenic fungal strains to infect Xylotrechus rusticus larvae. 3 Biotech, 2019, 9(3 80

[61] Wang YL, Lu Q, Decock C, Li YX, Zhang XY. Cytospora species from Populus and Salix in China with C. davidiana sp. nov. Fungal Biol, 2015, 119(5420-432

[62] Wu JH, Li JY, Zhang C, Yu XT, Cuthbertson AGS, Ali S. Biological impact and enzyme activities of Spodoptera litura (Lepidoptera: Noctuidae) in response to synergistic action of matrine and Beauveria brongniartii. Front Physiol, 2020, 11 584405

[63] Xie T, Jiang L, Li JS, Hong B, Wang XP, Jia YX. Effects of Lecanicillium lecanii strain JMC-01 on the physiology, biochemistry, and mortality of Bemisia tabaci Q-biotype nymphs. PeerJ, 2019, 7 e7690

[64] Yang YC, Ren LL, Xu LL, Wang T, Zong SX. Comparative morphology of sensilla on the antennae, maxillary and labial palps in different larval instars of Cryptorrhynchus lapathi (Linnaeus) (Coleoptera: Curculionidae). Zool Anz, 2019, 283: 93-101

[65] Yıldırım K, Eski A, Biryol S, Erdoğan P, Demir İ. Isolation, characterization, and formulation of indigenous Beauveria bassiana fungus against Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae). Potato Res, 2024, 67(2): 583-601

[66] Zeng T, Teng FY, Wei H, Lu YY, Xu YJ, Qi YX. Aanat1 regulates insect midgut detoxification through the ROS/CncC pathway. Commun Biol, 2024, 7(1): 808

[67] Zhang J, Song DL, Chen JX. Physiological and biochemical changes of the silkworm, Bombyx mori infected by Cordyceps militaris. Acta Entomol Sin, 2003, 46(6674-678

[68] Zhu CH, Sun JX, Tian FF, Tian XT, Liu Q, Pan YX, Zhang YJ, Luo ZB. The Bbotf1 Zn(II)₂Cys₆ transcription factor contributes to antioxidant response, fatty acid assimilation, peroxisome proliferation and infection cycles in insect pathogenic fungus Beauveria bassiana. J Invertebr Pathol, 2024, 204 108083

[69] Zou Y, Zhang LJ, Ge XZ, Guo SW, Li X, Chen LH, Wang T, Zong SX. Prediction of the long-term potential distribution of Cryptorhynchus lapathi (L.) under climate change. Forests, 2020, 11(1): 5

[70] Zuo TT, Wang MY, Zhang J, Li LM, Li DF, Zhang XJ. Antennal sensilla of Cryptorhynchus lapathi L. (Coleoptera: Curculionidae). Entomol News, 2015, 125(3199-213

RIGHTS & PERMISSIONS

Northeast Forestry University