Divergent responses of growth rate and antioxidative system of ten Bacillus strains to acid stresses

Highlights

● Response of growth rate and antioxidative system of ten Bacillus strains to acid stresses was assayed.

● Strong acid treatment significantly decreased the growth rate of the strains.

● Acid stresses increased the GPX activity and GSSG content of the tested strains.

● Divergent changes occurred in ROS and antioxidative system (SOD, CAT, GR, MDA and GSH).

Abstract

Environmental changes including soil acidification exert obvious stresses on soil ecosystems and influence soil microorganisms. In this study, ten microbial strains were incubated under different acid treatments to investigate responses of microbial growth and antioxidative system to acid stress. All the strains belong to Bacillus genus, but exhibit distinct ecological functions. We observed that these microbial strains had obviously different pH tolerance threshold, in spite of the close phylogenetic classification among strains. Acid stresses exerted significant effects on microbial antioxidative system, including superoxide dismutase (SOD), catalase (CAT) and glutathione transferring enzymes (GPX and GR) and reactants (GSH and GSSH), but the effects were strain specific. Furthermore, we found acid stress effects on total variances of the investigated microbial antioxidative system along the first two principal components (PCs). Activities of CAT and SOD contributed substantially to PC1 that reflected obvious acid effects on NC7 and ZC4, and closely related to intracellular malondialdehyde content. The GSSG activities and GSH/GSSG contributed greatly to PC2 that unveiled acid stress effects on most of the microbial strains. Our results highlight substantially heterogeneous responses of microbial strains to acid stress and support that phylogenetic closeness does not imply functional similarity of soil microorganisms under environmental changes.

Graphical abstract

Keywords

soil acidification / microbial strain / acid tolerance capacity / microbial growth / oxidative stress / antioxidative system

References

[1] Aguirre, J., Rios-Momberg, M., Hewitt, D., Hansberg, W., 2005. Reactive oxygen species and development in microbial eukaryotes. Trends in Microbiology13, 111–118.

[2] Alpert, A.J., Gilbert, H.F., 1985. Detection of oxidized and reduced glutathione with a recycling postcolumn reaction. Analytical Biochemistry144, 553–562.

[3] Alscher, R.G., Erturk, N., Heath, L.S., 2002. Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. Journal of Experimental Botany53, 1331–1341.

[4] Bahram, M., Hildebrand, F., Forslund, S.K., Anderson, J.L., Soudzilovskaia, N.A., Bodegom, P.M., Bengtsson-Palme, J., Anslan, S., Coelho, L.P., Harend, H., Huerta-Cepas, J., Medema, M.H., Maltz, M.R., Mundra, S., Olsson, P.A., Pent, M., Polme, S., Sunagawa, S., Ryberg, M., Tedersoo, L., Bork, P., 2018. Structure and function of the global topsoil microbiome. Nature560, 233–237.

[5] Branson, S.R., Broadbent, J.R., Carpenter, C.E., 2021. Internal pH and acid anion accumulation in Listeria monocytogenes and Escherichia coli exposed to lactic or acetic acids at mildly acidic pH. Frontiers in Microbiology12, 803271.

[6] Bruno-Bárcena, J.M., Azcárate-Peril, M.A., Hassan, H.M., 2010. Role of antioxidant enzymes in bacterial resistance to organic acids. Applied and Environmental Microbiology76, 2747–2753.

[7] Chowdhury, N., Goswami, G., Boro, R.C., Barooah, M., 2021. A pH-Dependent gene expression enables Bacillus amyloliquefaciens MBNC to adapt to acid stress. Current Microbiology78, 3104–3114.

[8] Crowther, T.W., van den Hoogen, J., Wan, J., Mayes, M.A., Keiser, A.D., Mo, L., Averill, C., Maynard, D.S., 2019. The global soil community and its influence on biogeochemistry. Science365, eaav0550.

[9] Dai, H., Wu, B., Chen, B., Ma, B., Chu, C., 2022. Diel fluctuation of extracellular reactive oxygen species production in the rhizosphere of rice. Environmental Science & Technology56, 9075–9082.

[10] Delgado-Baquerizo, M., Eldridge, D.J., Ochoa, V., Gozalo, B., Singh, B.K., Maestre, F.T., 2017. Soil microbial communities drive the resistance of ecosystem multifunctionality to global change in drylands across the globe. Ecology Letters20, 1295–1305.

[11] Fierer, N., 2017. Embracing the unknown: disentangling the complexities of the soil microbiome. Nature Reviews Microbiology15, 579–590.

[12] Gill, S.S., Tuteja, N., 2010. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry48, 909–930.

[13] Guan, N., Liu, L., 2020. Microbial response to acid stress: mechanisms and applications. Applied Microbiology and Biotechnology104, 51–65.

[14] Guan, S., 1986. Soil enzymes and the analytical methods. China Agriculture Press, Beijing

[15] Guo, J.H., Liu, X., Zhang, Y., Shen, J.L., Han, W.X., Zhang, W.F., Christie, P., Goulding, K.W.T., Vitousek, P.M., Zhang, F., 2010. Significant acidification in major Chinese croplands. Science327, 1008–1010.

[16] Hallbäcken, L., Tamm, C.O., 1986. Changes in soil acidity from 1927 to 1982–1984 in a forest area of south-west Sweden. Scandinavian Journal of Forest Research1, 219–232.

[17] Hao, T., Liu, X., Zhu, Q., Zeng, M., Chen, X., Yang, L., Shen, J., Shi, X., Zhang, F., de Vries, W., 2022. Quantifying drivers of soil acidification in three Chinese cropping systems. Soil & Tillage Research215, 105230.

[18] Hassan, Z.U., Al Thani, R., Alsafran, M., Migheli, Q., Jaoua, S., 2021. Selection of Bacillus spp. with decontamination potential on multiple Fusarium mycotoxins. Food Control127, 108119.

[19] Jamieson, D.J., 1998. Oxidative stress responses of the Yeast Saccharomyces cereisiae. Yeast (Chichester, England)14, 1511–1527.

[20] Jensen, L.E., Kragulund, L., Nybroe, O., 1998. Expression of a nitrogen regulated lux gene fusion in Pseudomonas fluorescens DF57 studied in pure culture and in soil. FEMS Microbiology Ecology25, 23–32.

[21] Joshi, S.G., Cooper, M., Yost, A., Paff, M., Ercan, U.K., Fridman, G., Friedman, G., Fridman, A., Brooks, A.D., 2011. Nonthermal dielectric-barrier discharge plasma-induced inactivation involves oxidative DNA damage and membrane lipid peroxidation in Escherichia coli. Antimicrobial Agents and Chemotherapy55, 1053–1062.

[22] Komives, T., Gullner, G., Király, Z., 1998. Role of glutathione and glutathione-related enzymes in response of plants to environmental stress. Annals of the New York Academy of Sciences851, 251–258.

[23] Lund, P., Tramonti, A., De Biase, D., 2014. Coping with low pH: molecular strategies in neutralophilic bacteria. FEMS Microbiology Reviews38, 1091–1125.

[24] Luo, Y., Vilain, S., Voigt, B., Albrecht, D., Hecker, M., Brozel, V.S., 2007. Proteomic analysis of Bacillus cereus growing in liquid soil organic matter. FEMS Microbiology Letters271, 40–47.

[25] Markewitz, D., Richter, D.D., Allen, H.L., Urrego, J.B., 1998. Three decades of observed soil acidification in the Calhoun Experimental Forest: Has acid rain made a difference?. Soil Science Society of America Journal62, 1428–1439.

[26] Mittler, R., 2002. Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science7, 405–410.

[27] Nilsson, J.F., Castellani, L.G., Draghi, W.O., Mogro, E.G., Wibberg, D., Winkler, A., Hansen, L.H., Schluter, A., Puhler, A., Kalinowski, J., Torres Tejerizo, G.A., Pistorio, M., 2021. Global transcriptome analysis of Rhizobium favelukesii LPU83 in response to acid stress. FEMS Microbiology Ecology97, fiaa235.

[28] Noctor, G., Gomez, L., Vanacker, H., Foyer, C.H., 2002. Interactions between biosynthesis, compartmentation and transport in the control of glutathione homeostasis and signalling. Journal of Experimental Botany53, 1283–1304.

[29] Nunan, N., Schmidt, H., Raynaud, X., 2020. The ecology of heterogeneity: soil bacterial communities and C dynamics. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences375, 20190249.

[30] Owens, C.W.I., Belcher, R.V., 1965. A colorimetric micro-method for the determination of glutathione. Biochemical Journal94, 705–711.

[31] Quivey Jr., R.G ., Faustoferri, R., Monahan, K., Marquis, R., 2000. Shifts in membrane fatty acid profiles associated with acid adaptation of Streptococcus mutans. FEMS Microbiology Letters189, 89–92.

[32] Puissant, J., Jones, B., Goodall, T., Mang, D., Blaud, A., Gweon, H.S., Malik, A., Jones, D.L., Clark, I.M., Hirsch, P.R., Griffiths, R., 2019. The pH optimum of soil exoenzymes adapt to long term changes in soil pH. Soil Biology and Biochemistry138, 107601.

[33] Raza, S., Miao, N., Wang, P., Ju, X., Chen, Z., Zhou, J., Kuzyakov, Y., 2020. Dramatic loss of inorganic carbon by nitrogen-induced soil acidification in Chinese croplands. Global Change Biology26, 3738–3751.

[34] Rousk, J., Baath, E., Brookes, P.C., Lauber, C.L., Lozupone, C., Caporaso, J.G., Knight, R., Fierer, N., 2010. Soil bacterial and fungal communities across a pH gradient in an arable soil. ISME Journal4, 1340–1351.

[35] Tullio, L.D., Gomes, D.F., Silva, L.P., Hungria, M., Batista, J.S.S., 2019. Proteomic analysis of Rhizobium freirei PRF 81(T) reveals the key role of central metabolic pathways in acid tolerance. Applied Soil Ecology135, 98–103.

[36] Vetrovsky, T., Kohout, P., Kopecky, M., Machac, A., Man, M., Bahnmann, B.D., Brabcova, V., Choi, J., Meszarosova, L., Human, Z.R., Lepinay, C., Llado, S., Lopez-Mondejar, R., Martinovic, T., Masinova, T., Morais, D., Navratilova, D., Odriozola, I., Stursova, M., Svec, K., Tlaskal, V., Urbanova, M., Wan, J., Zifcakova, L., Howe, A., Ladau, J., Peay, K.G., Storch, D., Wild, J., Baldrian, P., 2019. A meta-analysis of global fungal distribution reveals climate-driven patterns. Nature Communications10, 5142.

[37] Wang, Y., Li, J., Wang, J., Li, Z., 2010. Exogenous H₂O₂ improves the chilling tolerance of manilagrass and mascarenegrass by activating the antioxidative system. Plant Growth Regulation61, 195–204.

[38] Wei, H., Shan, X., Wu, L., Zhang, J., Saleem, M., Yang, J., Liu, Z., Chen, X., 2023. Microbial cell membrane properties and intracellular metabolism regulate individual level microbial responses to acid stress. Soil Biology & Biochemistry177, 108883.

[39] Wingsle, G., Hallgren, J.E., 1993. Influence of SO₂ and NO₂ exposure on glutathione, superoxide dismutase and glutathione reductase activties in Scots pine needles. Journal of Experimental Botany44, 463–470.

[40] Winterbourn, C.C., 2008. Reconciling the chemistry and biology of reactive oxygen species. Nature Chemical Biology4, 278–286.

[41] Wu, C., Huang, J., Zhou, R., 2014. Progress in engineering acid stress resistance of lactic acid bacteria. Applied Microbiology and Biotechnology98, 1055–1063.

[42] Wu, X.Y., von Tiedmeann, A., 2002. Impact of fungicides on active oxygen species and antioxidant enzymes in spring barley (Hordeum vulgare L. ) exposed to ozone. Environmental Pollution116, 37–47.

[43] Xu, Y., Zhao, Z., Tong, W., Ding, Y., Liu, B., Shi, Y., Wang, J., Sun, S., Liu, M., Wang, Y., Qi, Q., Xian, M., Zhao, G., 2020. An acid-tolerance response system protecting exponentially growing Escherichia coli. Nature Communications11, 1496.

[44] Yan, P., Wu, L., Wang, D., Fu, J., Shen, C., Li, X., Zhang, L., Zhang, L., Fan, L., Wenyan, H., 2020. Soil acidification in Chinese tea plantations. Science of the Total Environment715, 136963.

[45] Yang, Y., Ji, C., Ma, W., Wang, S., Wang, S., Han, W., Mohammat, A., Robinson, D., Smith, P., 2012. Significant soil acidification across northern China’s grasslands during 1980s-2000s. Global Change Biology18, 2292–2300.

[46] Yin, J., Chen, H., Duan, P., Zhu, K., Li, N., Ma, Y., Xu, Y., Guo, J., Liu, R., Chen, Q., 2023. Soil microbial communities as potential regulators of N₂O sources in highly acidic soils. Soil Ecology Letters5, 230178.

[47] Yoon, Y., Lee, H., Lee, S., Kim, S., Choi, K.H., 2015. Membrane fluidity-related adaptive response mechanisms of foodborne bacterial pathogens under environmental stresses. Food Research International72, 25–36.

[48] Yu, Z., Chen, H.Y.H., Searle, E.B., Sardans, J., Ciais, P., Peñuelas, J., Huang, Z., 2020. Whole soil acidification and base cation reduction across subtropical China. Geoderma361, 114107.

[49] Zhang, H., Huang, Y., An, S., Zeng, Q., Wang, B., Bai, X., Huang, Q., 2023. Decay stages and meteorological factors affect microbial community during leaf litter in situ decomposition. Soil Ecology Letters5, 220160.

[50] Zhou, Z., Wang, C., Luo, Y., 2020. Meta-analysis of the impacts of global change factors on soil microbial diversity and functionality. Nature Communications11, 3072.

Acknowledgments

This study is funded by the National Natural Science Foundation of China (Grant Nos. U1701236 and 32071641), the Joint Team Project of Guangdong Laboratory for Lingnan Modern Agriculture (Grant No. NT2021010), and Guangdong Science and Technology Department (Grant No. 2021A1515012507). We appreciate Prof. Yanfei Cai and Prof. Guixiang Peng for providing microbial strains and helps on strain culture.

Authors’ contribution

HW, XS and JZ conceived the experiment. XS, JC, ZL, and SC finished incubation and analyzed samples to collect data, analyzed and visualized data. XS and HW wrote the draft. JZ reviewed the manuscript and all the authors made significant contributions in result presentation. HW and JZ provided funding, project administration and supervision.

Competing interests

The authors declare no competing financial interests.

Data availability statement

All the data presented in this study are available by consulting to Dr. Hui Wei ([e-mail]).

Electronic supplementary material

Supplementary material is available in the online version of this article at https://doi.org/10.1007/s42832-023-0192-4 and is accessible for authorized users.

RIGHTS & PERMISSIONS

2023 Higher Education Press