Antimicrobial mechanism and biocontrol effect of Bacillus cereus XZ30-2 on Aspergillus niger

Main Article Content

Yanjie Yi
Zhipeng Hou
Qian Yang
Liuqing Cui
Heng Lu
Ruifang Li
Yang Liu
Yuyanqiao Zhang
Yuan Chen

Keywords

Aspergillus niger, Bacillus cereus XZ30-2, culture filtrate, antimicrobial mechanism, biocontrol effect

Abstract

Aspergillus niger is a major mold-causing spoilage in cereals, fruits and vegetables. Controlling of mold in stored grains is essential for safety of food. Currently, application of microorganisms to control A. niger is a safer and more effective method. In this study, strain XZ30-2 against A. niger was isolated and identified as Bacillus cereus according to morphological and biochemical characteristics as well as 16 Svedberg ribosomal ribonucleic acid (16S rRNA) gene sequence analysis. The investigation of action mechanism showed XZ30-2 culture filtrate caused the mycelia inflated or contract, increasing the membrane permeability, leading to the intracellular leakage and nucleic acids release, disrupting the proton pump, decreasing the ergosterol content, inducing the membrane lipid peroxidation and reactive oxygen species (ROS) accumulation in A. niger. Moreover, B. cereus XZ30-2 culture filtrate could produce hydrolases and lipopeptides, including iturin, surfactin and fengycin. This work also evaluated the control effect of XZ30-2 on A. niger in wheat grains, and indicated that 40 μL/g of culture filtrate significantly controlled the infection of A. niger. Therefore, B. cereus XZ30-2 can be developed as a biological agent for controlling A. niger in stored grains.

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References

Agriopoulou, S., Stamatelopoulou, E. and Varzakas, T., 2020. Advances in occurrence, importance, and mycotoxin control strategies: prevention and detoxification in foods. Foods 9(2): 137. 10.3390/foods9020137

Alkuwari, A., Hassan, Z.U., Zeidan, R., Al-Thani, R. and Jaoua, S., (2022) Occurrence of mycotoxins and toxigenic fungi in cereals and application of Yeast volatiles for their biological control. Toxins (Basel), 14(6). 10.3390/toxins14060404

Ayangbenro, A.S. and Babalola, O.O., 2020. Genomic analysis of Bacillus cereus NWUAB01 and its heavy metal removal from polluted soil. Scientific Reports 10(1): 19660. 10.1038/s41598-020-75170-x

Báez-Astorga, P.A., Cázares-Álvarez, J.E., Cruz-Mendívil, A., Quiroz-Figueroa, F.R., Sánchez-Valle, V.I. and Maldonado-Mendoza, I.E., 2022. Molecular and biochemical characterisation of antagonistic mechanisms of the biocontrol agent Bacillus cereus B25 inhibiting the growth of the phytopathogen Fusarium verticillioides P03 during their direct interaction in vitro. Biocontrol Science and Technology 32(9): 1074–1094. 10.1080/09583157.2022.2085662

Ben Khedher, S., Mejdoub-Trabelsi, B. and Tounsi, S., 2021. Biological potential of Bacillus subtilis V26 for the control of Fusarium wilt and tuber dry rot on potato caused by Fusarium species and the promotion of plant growth. Biological Control 152: 104444. 10.1016/j.biocontrol.2020.104444

de Andrade Santiago, J., Cardoso, M.D.G., Batista, L.R., Santiago, W.D., Passamani, F.R.F., Rodrigues, L.M.A. and Nelson, D.L., 2018. Effect of the essential oils from Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on the synthesis of ochratoxin A by Aspergillus niger and Aspergillus carbonarius isolated from tropical wine grapes. Journal of Food Science and Technology-Mysore 55(1): 418–423. 10.1007/s13197-017-2857-4

Duan, W., Zhang, S., Lv, Y., Zhai, H., Wei, S., Ma, P., Cai, J. and Hu, Y., 2023. Inhibitory effect of (E)-2-heptenal on Aspergillus flavus growth revealed by metabolomics and biochemical analyses. Applied Microbiology and Biotechnology 107(1): 341–354. 10.1007/s00253-022-12320-3

Durval, I.J.B., Meira, H.M., de Veras, B.O., Rufino, R.D., Converti, A. and Sarubbo, L.A., 2021. Green synthesis of silver nanoparticles using a biosurfactant from Bacillus cereus UCP 1615 as stabilizing agent and its application as an antifungal agent. Fermentation (Basel) 7(4): 233. 10.3390/fermentation7040233

Fleurat-Lessard, F., 2017. Integrated management of the risks of stored grain spoilage by seedborne fungi and contamination by storage mould mycotoxins–an update. Journal of Stored Products Research 71: 22–40. 10.1016/j.jspr.2016.10.002

Frisvad, J.C., Moller, L.L.H., Larsen, T.O., Kumar, R. and Arnau, J., 2018. Safety of the fungal workhorses of industrial biotechnology: update on the mycotoxin and secondary metabolite potential of Aspergillus niger, Aspergillus oryzae, and Trichoderma reesei. Applied Microbiology and Biotechnology 102(22): 9481–9515. 10.1007/s00253-018-9354-1

Georgopapadakou, N.H. and Tkacz, J.S., 1995. The fungal cell wall as a drug target. Trends in Microbiology 3(3): 98–104. 10.1016/S0966-842X(00)88890-3

Gherbawy, Y.A., Maghraby, T.A., Hamza, L.H.A. and El-Dawy, E., 2021. New morphological criteria and molecular characterization of black aspergilli aggregate from corn, sorghum and wheat grains. Archives of Microbiology 203(1): 355–366. 10.1007/s00203-020-02024-5

Gil-Serna, J., Garcia-Diaz, M., Vazquez, C., Gonzalez-Jaen, M.T. and Patino, B., 2019. Significance of Aspergillus niger aggregate species as contaminants of food products in Spain regarding their occurrence and their ability to produce mycotoxins. Food Microbiology 82: 240–248. 10.1016/j.fm.2019.02.013

He, R., Yang, Y., Hu, Z., Xue, R. and Hu, Y., 2021. Resistance mechanisms and fitness of pyraclostrobin-resistant isolates of Lasiodiplodia the obromae from mango orchards. PLoS One 16(6): e0253659. 10.1371/journal.pone.0253659

Hernandez-Huerta, J., Tamez-Guerra, P., Gomez-Flores, R., Delgado-Gardea, M.C.E., Robles-Hernandez, L., Gonzalez-Franco, A.C. and Infante-Ramirez, R., 2023. Pepper growth promotion and biocontrol against Xanthomonas euvesicatoria by Bacillus cereus and Bacillus thuringiensis formulations. PeerJ 11: e14633. 10.7717/peerj.14633

Hu, J., Dong, B., Wang, D., Meng, H., Li, X. and Zhou, H., 2023. Genomic and metabolic features of Bacillus cereus inhibiting the growth of Sclerotinia sclerotiorum by synthesizing secondary metabolites. Archives of Microbiology 205(1): 1–13. 10.1007/s00203-022-03351-5

Ju, J., Lei, Y., Guo, Y., Yu, H., Cheng, Y. and Yao, W., 2023. Eugenol and citral kills Aspergillus niger through the tricarboxylic acid cycle and its application in food preservation. Food Science and Technology (LWT) 173: 114226. 10.1016/j.lwt.2022.114226

Ju, J., Xie, Y., Yu, H., Guo, Y., Cheng, Y., Zhang, R. and Yao, W., 2020. Synergistic inhibition effect of citral and eugenol against Aspergillus niger and their application in bread preservation. Food Chemistry 310: 125974. 10.1016/j.foodchem.2019.125974

Ke, Y., Ding, B., Zhang, M., Dong, T., Fu, Y., Lv, Q., Ding, W. and Wang, X ., 2022. Study on inhibitory activity and mechanism of chitosan oligosaccharides on Aspergillus Flavus and Aspergillus Fumigatus. Carbohydrate Polymers 275: 118673. 10.1016/j.carbpol.2021.118673

Li, T., Li, L., Du, F., Sun, L., Shi, J., Long, M., and Chen, Z., 2021. Activity and mechanism of action of antifungal peptides from microorganisms: a review. Molecules 26(11): 3438. 10.3390/molecules26113438

Li, S., Xu, X., Zhao, T., Ma, J., Zhao, L., Song, Q. and Sun, W., 2022. Screening of Bacillus velezensis E2 and the inhibitory effect of its antifungal substances on Aspergillus flavus. Foods 11(2): 140. 10.3390/foods11020140

Li, F., Zeng, Y., Zong, M., Yang, J. and Lou, W., 2020. Bioprospecting of a novel endophytic Bacillus velezensis FZ06 from leaves of Camellia assamica: production of three groups of lipopeptides and the inhibition against food spoilage microorganisms. Journal of Biotechnology 323: 42–53. 10.1016/j.jbiotec.2020.07.021

Mannaa, M. and Kim, K.D., 2017. Influence of temperature and water activity on deleterious fungi and mycotoxin production during grain storage. Mycobiology 45(4): 240–254. 10.5941/MYCO.2017.45.4.240

Mesterházy, Á., Oláh, J. and Popp, J., 2020. Losses in the grain supply chain: causes and solutions. Sustainability 12(6): 2342. 10.3390/su12062342

Ming, S., Chen, X., Zhang, N., Li, S., Zhu, Z. and Cheng, S., 2022. Structure and stability analysis of antibacterial substance produced by selenium enriched Bacillus cereus BC1. Archives of Microbiology 204(3): 196. 10.1007/s00203-022-02798-w

Molina-Hernández, J.B., Scroccarello, A., Della Pelle, F., De Flaviis, R., Compagnone, D., Del Carlo, M. and Lόpez, C. C., 2022. Synergistic antifungal activity of catechin and silver nanoparticles on Aspergillus niger isolated from coffee seeds. Food Science and Technology (LWT) 169: 113990. 10.1016/j.lwt.2022.113990

Niu, A., Wu, H., Ma, F., Tan, S., Wang, G. and Qiu, W., 2022. The antifungal activity of cinnamaldehyde in vapor phase against Aspergillus niger isolated from spoiled paddy. Food Science and Technology (LWT) 159: 113181. 10.1016/j.lwt.2022.113181

Nxumalo, C.I., Ngidi, L.S., Shandu, J.S.E. and Maliehe, T.S., 2020. Isolation of endophytic bacteria from the leaves of Anredera cordifolia CIX1 for metabolites and their biological activities. BMC Complementary Medicine and Therapies 20(1): 300. 10.1186/s12906-020-03095-z

Oztopuz, O., Pekin, G., Park, R.D. and Eltem, R., 2018. Isolation and evaluation of new antagonist Bacillus strains for the control of pathogenic and mycotoxigenic fungi of fig orchards. Applied Biochemistry and Biotechnology 186(3): 692–711. 10.1007/s12010-018-2764-9

Ponsone, M.L., Chiotta, M.L., Combina, M., Dalcero, A. and Chulze, S., 2011. Biocontrol as a strategy to reduce the impact of ochratoxin A and Aspergillus section Nigri in grapes. International Journal of Food Microbiology 151(1): 70–77. 10.1016/j.ijfoodmicro.2011.08.005

Prakash, B., Singh, P., Goni, R., Raina, A.K.P. and Dubey, N.K., 2014. Efficacy of Angelica archangelica essential oil, phenyl ethyl alcohol and α-terpineol against isolated molds from walnut and their antiaflatoxigenic and antioxidant activity. Journal of Food Science and Technology 52(4): 2220–2228. 10.1007/s13197-014-1278-x

Rahayu, E.S., Triyadi, R., Khusna, R.N.B., Djaafar, T.F., Utami, T., Marwati, T. and Hatmi, R. U., 2021. Indigenous yeast, lactic acid bacteria and acetic acid bacteria from cocoa bean fermentation in Indonesia can inhibit fungal-growth-producing mycotoxins. Fermentation (Basel ) 7(3): 192. 10.3390/fermentation7030192

Rani, M., Weadge, J.T. and Jabaji, S., 2020. Isolation and characterization of biosurfactant-producing bacteria from oil well batteries with antimicrobial activities against food-borne and plant pathogens. Frontiers in Microbiology 11: 64. 10.3389/fmicb.2020.00064

Rutenberg, R., Bernstein, S., Fallik, E., Paster, N. and Poverenov, E., 2018. The improvement of propionic acid safety and use during the preservation of stored grains. Crop Protection 110: 191–197. 10.1016/j.cropro.2017.09.005

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. 10.1093/oxfordjournals.molbev.a040454

Santra, H.K. and Banerjee, D., 2020. Natural products as fungicide and their role in crop protection. Springer, Singapore. 10.1007/978-981-15-3024-1_9

Tao, N., OuYang, Q. and Jia, L., 2014. Citral inhibits mycelial growth of Penicillium italicum by a membrane damage mechanism. Food Control 41: 116–121. 10.1016/j.foodcont.

2014.01.010

Thorpe, G.R., 2008. The application of computational fluid dynamics codes to simulate heat and moisture transfer in stored grains. Journal of Stored Products Research 44(1): 21–31. 10.1016/j.jspr.2007.07.001

Tian, J., Ban, X., Zeng, H., He, J., Chen, Y. and Wang, Y., 2012. The mechanism of antifungal action of essential oil from dill (Anethum graveolens L.) on Aspergillus flavus. Plos One, 7(1): e30147. 10.1016/j.foodchem.2011.07.061

Tian, J., Huang, B., Luo, X., Zeng, H., Ban, X., He, J. and Wang, Y., 2012. The control of Aspergillus flavus with Cinnamomum jensenianum Hand.-Mazz essential oil and its potential use as a food preservative. Food Chemistry 130(3): 520–527. 10.1016/j.foodchem.2011.07.061

Tso, K.H., Lumsangkul, C., Ju, J., Fan, Y. and Chiang, H.I., 2021. The potential of peroxidases extracted from the spent mushroom (Flammulina velutipes) substrate significantly degrade mycotoxin deoxynivalenol. Toxins (Basel) 13(1): 72. 10.3390/toxins13010072

Ul Hassan, Z., Al Thani, R., Alnaimi, H., Migheli, Q. and Jaoua, S., 2019. Investigation and application of Bacillus licheniformis volatile compounds for the biological control of toxigenic Aspergillus and Penicillium spp. ACS Omega 4(17): 17186–17193. 10.1021/acsomega.9b01638

van Schie, L., Borgers, K., Michielsen, G., Plets, E., Vuylsteke, M., Tiels, P., Festjens, N. and Callewaert, N., 2021. Exploration of synergistic action of cell wall-degrading enzymes against mycobacterium tuberculosis. Antimicrobial Agents and Chemotherapy 65(10): e00659–00621. 10.1128/AAC.00659-21

Vos, P., Garrity, G.M., Jones D., Krieg, N.R., Ludwig, W., Rainey, F.A., Schleifer KH. and Whitman, W.B. (Eds.). 2009 Bergey’s manual of systematic bacteriology, 2nd ed. Springer, New York, NY.

Wang, Q., Lin, Q., Peng, K., Cao, J., Yang, C. and Xu, D., 2017. Surfactin variants from Bacillus subtilis natto CSUF5 and their antifungal properities against Aspergillus niger. Journal of Biobased Materials and Bioenergy 11(3): 210–215. 10.1166/jbmb.2017.1665

Wang, Y., Zhang, J., Wang, Y., Wang, K., Wei, H. and Shen, L., 2018. Isolation and characterization of the Bacillus cereus BC7 strain, which is capable of zearalenone removal and intestinal flora modulation in mice. Toxicon 155: 9–20. 10.1016/j.toxicon.2018.09.005

Wilson, D.M., Mubatanhema, W. and Jurjevic, Z., 2002. Biology and ecology of mycotoxigenic Aspergillus species as related to economic and health concerns. Advances in Experimental Medicine and Biology 504: 3–17. 10.1007/978-1-4615-0629-4_2

Yan, H., Meng, X., Lin, X., Duan, N., Wang, Z. and Wu, S., 2023. Antifungal activity and inhibitory mechanisms of ferulic acid against the growth of Fusarium graminearum. Food Bioscience 52: 102414. 10.1016/j.fbio.2023.102414

Yan, P., Zhang, X., Hu, L., Wang, Y., Zhu, M.L., Wu, X. and Chen, F., 2020. Two novel strains, Bacillus albus JK-XZ3 and B. velezensis JK-XZ8, with activity against Cerasus crown gall disease in Xuzhou, China. Australasian Plant Pathology 49(2): 127–136. 10.1007/s13313-020-00682-z

Yassein, A.S. and Elamary, R.B., 2021. Efficacy of soil Paraburkholderia fungorum and Bacillus subtilis on the inhibition of Aspergillus niger growth and its ochratoxins production. Egyptian Journal of Botany 61(1): 319–334. 10.21608/ejbo.2021.68481.1656

Yi, Y., Luan, P., Liu, S., Shan, Y., Hou, Z., Zhao, S., Jia, S. and Li, R., 2022. Efficacy of Bacillus subtilis XZ18-3 as a biocontrol agent against Rhizoctonia cerealis on wheat. Agriculture 12(2): 258. 10.3390/agriculture12020258

Zhai, Y., Zhu, J., Tan, T., Xu, J., Shen, A., Yang, X., Li, J., Zeng, L., and Wei, L., 2021. Isolation and characterization of antagonistic Paenibacillus polymyxa HX-140 and its biocontrol potential against Fusarium wilt of cucumber seedlings. BMC Microbiology 21(1): 75. 10.1186/s12866-021-02131-3

Zhang, D., Qiang, R., Zhou, Z., Pan, Y., Yu, S., Yuan, W., Cheng, J., Wang, J., Zhao, D., Zhu, J. and Yang, Z., 2022a. Biocontrol and action mechanism of Bacillus subtilis lipopeptides’ fengycins against alternaria solani in potato as assessed by a transcriptome analysis. Frontiers in Microbiology 13: 861113. 10.3389/fmicb.2022.861113

Zhang, R.X., Wu, Z.W., Cui, H.Y., Chai, Y.N., Hua, C.W., Wang, P., et al., 2022b. Production of surfactant-stable keratinase from Bacillus cereus YQ15 and its application as detergent additive. BMC Biotechnology 22(1): 1–13. 10.1186/s12896-022-00757-3