Published date: Jun 07 2022

Journal Title: KnE Life Sciences

Issue title: The First Asian PGPR Indonesian Chapter International e-Conference 2021

Pages: 648–663

DOI: 10.18502/kls.v7i3.11169

Yuda Purwana Roswanjayayuda.purwana.roswanjaya@brin.go.idResearch Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Indonesia

Nur Alfi SaryanahResearch Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Indonesia

Lukita DevyResearch Center for Horticultural and Estate Crops, National Research and Innovation Agency (BRIN), Indonesia

Beauveria bassiana is an important entomopathogenic fungus that is widely used as a bioinsecticide around the world. Conidia production is a critical step in the production of high-quality bioinsecticide. This study investigated three liquid culture mediums and five combinations of solid substrates to enhance conidia production by B. bassiana. The fungus was isolated from infected insects in the cocoa plantation of PT. Perkebunan Nusantara XII in Kediri, East Java, Indonesia. The three culture mediums were malt extract broth (MB), potato dextrose broth (PDB), and yeast and malt extract broth (YMB). Five combinations of solid substrate were used: 100% rice, 100% maize, 75%:25% rice:maize, 50%:50% rice:maize, and 25%:75% rice:maize. The biphasic system was used in this study, in which the fungus was first grown under submerged conditions and then was allowed to conidiate in solid-state conditions. The data showed that PDB was the optimum culture medium to produce blastophore and beauvericin, the active compound that acts as a mycoinsecticide. In the selection test, 100% rice was the optimum solid substrate to produce high amounts of conidia, and the consistency and production tests yielded the same results, with conidia counts of 1.93x109, 1.78x109, and 2.08x109, respectively. In a rice storability test, B. bassiana conidia numbers remained stable for up to 105 days of storage at room temperature.

Keywords: Beauveria bassiana, culture medium, solid-substrate, conidia, biphasic system

[1] Naqqash MN, Gökçe A, Bakhsh A, Salim M. Insecticide resistance and its molecular basis in urban insect pests. Parasitology Research. 2016;115(4):1363–1373.

[2] Umina PA, McDonald G, Maino J, Edwards O, Hoffmann AA. Escalating insecticide resistance in Australian grain pests: Contributing factors, industry trends and management opportunities. Pest Management Science. 2019;75(6):1494–1506.

[3] Kim K-H, Kabir E, Jahan SA. Exposure to pesticides and the associated human health effects. Science of The Total Environment. 2017;575:525–535.

[4] Sabarwal A, Kumar K, Singh RP. Hazardous effects of chemical pesticides on human health – Cancer and other associated disorders. Environmental Toxicology and Pharmacology. 2018;63:103–114.

[5] Keyhani NO. Lipid biology in fungal stress and virulence: Entomopathogenic fungi. Fungal Biology. 2018;122(6):420–429.

[6] Olson S. An analysis of the biopesticide market now and where it is going. Outlooks on Pest Management. 2015;26(5):203–206.

[7] Damalas CA, Koutroubas SD. Current status and recent developments in biopesticide use. Agriculture. 2018;8(1):1-6.

[8] Arthurs S, Dara SK. Microbial biopesticides for invertebrate pests and their markets in the United States. Journal of Invertebrate Pathology. 2019;165:13–21.

[9] Vega FE, Goetell MS, Blackwell M et al. Fungal entomopathogens: New insights on their ecology. Fungal Ecology. 2009;2(4):149–159.

[10] Garrido-Jurado I, Marquez M, Ortiz-Urquiza A et al. Genetic analyses place most Spanish isolates of Beauveria bassiana in a molecular group with word-wide distribution. BMC Microbiology. 2011;11(1):84–94.

[11] McKinnon AC, Saari S, Moran-Diez ME, Meyling NV, Raad M, Glare TR. Beauveria bassiana as an endophyte: A critical review on associated methodology and biocontrol potential. BioControl. 2017;62(1):1–17.

[12] Ownley BH, Griffin MR, Klingeman WE, Gwinn KD, Moulton JK, Pereira RM. Beauveria bassiana: Endophytic colonization and plant disease control. Journal of Invertebrate Pathology. 2008;98(3):267–270.

[13] Quesada-Moraga E, López-Díaz C, Landa BB. The hidden habit of the entomopathogenic fungus Beauveria bassiana: First demonstration of vertical plant transmission. PLOS ONE. 2014;9(2):e89278– e892713

[14] Garrido-Jurado I, Fernández-Bravo M, Campos C, Quesada-Moraga E. Diversity of entomopathogenic hypocreales in soil and phylloplanes of five mediterranean cropping systems. Journal of Invertebrate Pathology. 2015;130:97–106.

[15] Mascarin GM, Jaronski ST. The production and uses of Beauveria bassiana as a microbial insecticide. World Journal of Microbiology and Biotechnology. 2016;32(11):177–202.

[16] Hegedus DD, Bidochka MJ, Miranpuri GS, Khachatourians GG. A comparison of the virulence, stability and cell-wall-surface characteristics of three spore types produced by the entomopathogenic fungus Beauveria bassiana. Applied Microbiology and Biotechnology. 1992;36(6):785–789.

[17] Lopez-Perez M, Rodriguez-Gomez D, Loera O. Production of conidia of Beauveria bassiana in solid-state culture: Current status and future perspectives. Critical Reviews in Biotechnology. 2015;35(3):334–341.

[18] Mascarin GM, Jackson MA, Behle RW, Kobori NN, Júnior ID. Improved shelf life of dried Beauveria bassiana blastospores using convective drying and active packaging processes. Applied Microbiology and Biotechnology. 2016;100(19):8359– 8370.

[19] Roswanjaya YP, Saryanah NA, Nawfetrias W, Rosdayanti H, Putri AL. Selection of culture medium and incubation time for growth and production of beauvericin by local Beauveria bassiana. Advances in Biological Sciences Research 2021;13(5):59– 65.

[20] Pham TA, Kim JJ, Mm SG, Kim K. Production of blastospore of entomopathogenic Beauveria bassiana in a submerged batch culture. Mycobiology. 2009;37(3):218– 224.

[21] Puzari KC, Sarmah DK, Hazarika LK. Medium for mass production of Beauveria bassiana (balsamo) vuillemin. Journal of Biological Control. 1997;11(1&2): 97–100.

[22] Xie L, Chen HM, Yang JB. Conidia production by Beauveria bassiana on rice in solidstate fermentation using tray bioreactor. Advanced Materials Research. 2013;610– 613:3478–3482.

[23] Lacey LA. Manual of techniques in insect pathology. Academic Press, Wapato USA; 1997.

[24] Moretti A, Logrieco A, Bottalico A, Ritieni A, Randazzo G, Corda P. Beauvericin production by Fusarium subglutinans from different geographical areas. Mycological Research. 1995;99(3):282–286.

[25] Antonio L, Antonio M, Ritieni A et al. Natural occurrence of beauvericin in preharvest Fusarium subglutinans infected corn ears in Poland. Journal of Agricultural and Food Chemistry. 1993;41(11):2149–2152.

[26] Pham TA, Kim JJ, Kim K. Selection of solid-state fermentation for improved conidia production of Beauveria bassiana as a mycoinsecticide. Mycobiology. 2010;38(2):137–143.

[27] Posada-Flórez FJ. Production of Beauveria bassiana fungal spores on rice to control the coffee berry borer, Hypothenemus hampei, in Colombia. Journal of Insect Science. 2008;8(1): 1–13.

[28] Iskandarov US, Guzalova AG, Davranov KD. Effects of nutrient medium composition and temperature on the germination of conidia and the entomopathogenic activity of the fungi Beauveria bassiana and Metarhizium anisopliae. Applied Biochemistry and Microbiology. 2006;42(1):72–76.

[29] Strasser H, Vey A, Butt TM. Are there any risks in using entomopathogenic fungi for pest control, with particular reference to the bioactive metabolites of Metarhizium, Tolypocladium and Beauveria species? Biocontrol Science and Technology. 2000;10(6):717–735.

[30] Lee HS, Song HH, Ahn JH, Shin CG, Lee GP, Lee C. Statistical selection of growth medium for the production of the entomopathogenic and phytotoxic cyclic depsipeptide beauvericin from Fusarium oxysporum KFCC 11363P. Journal of Microbiology and Biotechnology. 2008;18(1):138–144.

[31] Jenkins NE, Heviefo G, Langewald J, Lomer CJ. Development of mass production technology for aerial conidia for use as mycopesticides. Biocontrol News and Information 1998;19(1):21–32

[32] Tarocco F, Lecuona RE, Couto AS, Arcas JA. Selection of erythritol and glycerol accumulation in conidia of Beauveria bassiana by solid-state fermentation, using response surface methodology. Applied Microbiology and Biotechnology. 2005;68(4):481–488.

[33] Ye SD, Ying SH, Chen C, Feng MG. New solid-state fermentation chamber for bulk production of aerial conidia of fungal biocontrol agents on rice. Biotechnology Letter. 2006;28(11):799–804.

[34] Kreutz J, Vaupel O, Zimmermann G. Efficacy of Beauveria bassiana (Bals.) vuill. against the spruce bark beetle, Ips Typographus L., in the laboratory under various conditions. Journal of Applied Entomology. 2004;128(6):384–389.

[35] Latifian M, Rad B, Amani M, Rahkhodaei E. Mass production of entomopathogenic fungi Beauveria bassiana (Balsamo) by using agricultural products based on liquidsolid diphasic method for date palm pest control. International Journal of Agriculture and Crop Sciences. 2013;19:2337–2341.

[36] Figueroa-Montero A, Esparza-Isunza T, Saucedo-Castañeda G, Huerta-Ochoa S, Gutiérrez-Rojas M, Favela-Torres E. Improvement of heat removal in solid-state fermentation tray bioreactors by forced air convection. Journal of Chemical Technology & Biotechnology. 2011;86(10):1321–1331.

[37] Membrillo I, Sánchez C, Meneses M, Favela E, Loera O. Particle geometry affects differentially substrate composition and enzyme profiles by Pleurotus ostreatus growing on sugar cane bagasse. Bioresource Technology. 2011;102(2):1581–1586.

[38] Sy VE, Schalamuk S, Scorsetti AC, Botto IL. Volcanic materials as carriers for the formulation of mycoinsecticides using the fungus Beauveria bassiana. International Journal of Agriculture and Natural Resources. 2016;43(2): 273–282.

[39] Kim JC, Lee MR, Kim S et al. Long-term storage stability of Beauveria bassiana ERL836 granules as fungal biopesticide. Journal of Asia-Pacific Entomology. 2019;22(2):537–542.

[40] Villamizar LF, Nelson TL, Jones SA, Jackson TA, Hurst HRK, Marshall SDG. Formation of microsclerotia in three species of Beauveria and storage stability of a prototype granular formulation. Biocontrol Science and Technology. 2018;28(12):1097–1113.

[41] Faria M, Lopes RB, Souza DA, Wraight SP. Conidial vigor vs. viability as predictors of virulence of entomopathogenic fungi. Journal of Invertebrate Pathology. 2015;125:68–72.

[42] Faria M, Hotchkiss JH, Wraight SP. Application of modified atmosphere packaging (gas flushing and active packaging) for extending the shelf life of Beauveria bassiana conidia at high temperatures. Biological Control. 2012;61(1):78–88.