Impact of photoperiod and soybean genotypes (E-genes) on the composition of root exudates, growth and biofilm formation of rhizosphere microbiota of soybean isogenic lines

D. V. Hlushach; O. O. Avksentieva; Y. G. Kot

doi:10.15421/0225079

D. V. Hlushach V. N. Karazin Kharkiv National University
O. O. Avksentieva V. N. Karazin Kharkiv National University
Y. G. Kot V. N. Karazin Kharkiv National University

Keywords: Glycine max, rhizobacteria, photoperiod sensitivity, biofilm, root secretion, flavonoids, phenolic compounds, amino acids, carbohydrate.

Abstract

The study is dedicated to analyzing the impact of photoperiod on the quantitative and qualitative composition of root exudates in Glycine max (L.) Merr. with varying photoperiod sensitivity, as well as their influence on the growth and biofilm formation of the soil bacteria Bradyrhizobium japonicum and Bacillus subtilis . Soybean near-isogenic lines (NILs) differing in the allelic state of photoperiod sensitivity genes ( E1–E3 ) were used. The p lants were grown for 14 days in a controlled environment chamber under two light conditions: a short day (9 hours) and a long day (16 hours). Root exudates were collected for biochemical analysis of carbohydrate, protein, amino acid, phenol, and flavonoid content, and to evaluate their effects on bacterial growth and biofilm formation. The results demonstrate that the photoperiod significantly affects the composition of root exudates, with the effect depending on the allelic state of the E genes in the plant genotype. Under short-day conditions, short-day lines (Clark and L80-5879) and the day-neutral line L63-3117 exhibited an increase in monosaccharide content. In root exudates of all lines, short days led to a decrease in soluble protein content, and in lines with dominant E1–E3 genes, a reduction in amino acid content was observed. An exception was the day-neutral line L71-920 ( e1e2e3 ), in which amino acid content increased under short-day conditions. Short days significantly increased phenol content in the exudates of L63-3117 but decreased it in L71-920. In lines with the dominant E3 gene (Clark and L63-3117), short days led to an increase in flavonoid content, while in other lines it decreased. R oot exudates of the cv. Clark under short-day conditions inhibited the growth of B. japonicum , likely due to reduced amino acid and protein content, which serve as nitrogen sources, and an increase in flavonoids, which may exert an inhibitory effect. Exudates from line L80-5879 did not affect rhizobial growth, while those from L63-3117 inhibited it, likely due to low amino acid content. In contrast, exudates of L71-920 under short days stimulated rhizobial growth, which correlated with increased amino acid content, potentially acting as chemoattractants. Biofilm formation by Br. japonicum was suppressed by exudates of short-day lines regardless of photoperiod, whereas in day-neutral lines (L63-3117 and L71-920), short-day conditions significantly stimulated biofilm formation. For B. subtilis , exudates of all lines under long-day conditions inhibited biofilm formation, but under short days, lines with dominant E1–E3 genes promoted it, likely due to increased monosaccharide content as a carbon source. In co-cultivation of rhizobia and bacilli, exudates from most lines under short days enhanced biofilm formation, except in L71-920, where a decrease in monosaccharide content was observed. These results highlight the importance of gen o typic photoperiod sensitivity in forming the composition of root exudates and their influence on the rhizosphere microbiota in response to photoperiod. This has important implications for understanding the mechanisms regulating symbiotic interactions between soybean and microorganisms, and for optimizing agronomic practices for soybean cultivation across different latitudes.

References

Atlas, R. M. (2005). Handbook of media for environmental microbiology. 2nd ed. CRC Press, Boca Raton.

Badri, D. V., & Vivanco, J. M. (2009). Regulation and function of root exudetes. Plant, Cell and Environment, 32(6), 666–681.

Barbour, W. M., Hattermann, D. R., & Stacey, G. (1991). Chemotaxis of Bradyrhizobium japonicum to soybean exudates. Applied and Environmental Microbiology, 57(9), 2635–2639.

Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1–2), 248–254.

Canarini, A., Kaiser, C., Merchant, A., Richter, A., & Wanek, W. (2019). Root exudation of primary metabolites: Mechanisms and their roles in plant responses to environmental stimuli. Frontiers in Plant Science, 10, 157.

Cesari, A., Paulucci, N., López-Gómez, M., Hidalgo-Castellanos, J., Plá, C. L., & Dardanelli, M. S. (2019). Restrictive water condition modifies the root exudates composition during peanut-PGPR interaction and conditions early events, reversing the negative effects on plant growth. Plant Physiology and Biochemistry, 142, 519–527.

Chamam, A., Sanguin, H., Bellvert, F., Meiffren, G., Comte, G., Wisniewski-Dyé, F., Bertrand, C., & Prigent-Combaret, C. (2013). Plant secondary metabolite profiling evidences strain-dependent effect in the Azospirillum–Oryza sativa association. Phytochemistry, 87, 65–77.

Chatterton, N. J., & Silvius, J. E. (1979). Photosynthate partitioning into starch in soybean leaves. Plant Physiology, 64(5), 749–753.

Chatterton, N. J., & Silvius, J. E. (1980). Acclimation of photosynthate partitioning and photosynthetic rates to changes in length of the daily photosynthetic period. Annals of Botany, 46(6), 739–745.

Chuiko, N. V., Antonyuk, T. S., & Kurdish, I. K. (2002). The chemotactic response of Bradyrhizobium japonicum to various organic compounds. Microbiology, 71(4), 391–396.

Coronado, C., Zuanazzi, J. A. S., Sallaud, C., Quirion, J. C., Esnault, R., Husson, H. P., Kondorosi, A., & Ratet, P. (1995). Alfalfa root flavonoid production is nitrogen regulated. Plant Physiology, 108(2), 533–542.

Danhorn, T., & Fuqua, C. (2007). Biofilm formation by plant-associated bacteria. Annual Review of Microbiology, 61, 401–422.

Du, J., Li, Y., Ur-Rehman, S., Mukhtar, I., Yin, Z., Dong, H., Wang, H., Zhang, X., Gao, Z., Zhao, X., Xin, X., & Ding, X. (2021). Synergistically promoting plant health by harnessing synthetic microbial communities and prebiotics. iScience, 24(8), 102918.

Fattahi, S., Zabihi, E., Abedian, Z., Pourbagher, R., Motevalizadeh Ardekani, A., Mostafazadeh, A., & Akhavan-Niaki, H. (2014). Total phenolic and flavonoid contents of aqueous extract of stinging nettle and in vitro antiproliferative effect on HeLa and BT-474 cell lines. International Journal of Molecular and Cellular Medicine, 3(2), 102–107.

Gage, D. J. (2004). Infection and invasion of roots by symbiotic, nitrogen-fixing rhizobia during nodulation of temperate legumes. Microbiology and Molecular Biology Reviews, 68(2), 280–300.

Gendron, J. M., & Staiger, D. (2023). New horizons in plant photoperiodism. Annual Review of Plant Biology, 74(1), 481–509.

Guo, M., Yang, G., Meng, X., Zhang, T., Li, C., Bai, S., & Zhao, X. (2023). Illuminating plant–microbe interaction: How photoperiod affects rhizosphere and pollutant removal in constructed wetland? Environment International, 179, 108144.

Hayat, R., Ahmed, I., & Sheirdil, R. A. (2012). An overview of plant growth promoting rhizobacteria (PGPR) for sustainable agriculture. In: Ashraf, M., Öztürk, M., Ahmad, M. S. A., & Aksoy, A. (Eds.). Crop production for agricultural improvement. Springer, Dordrecht. Pp. 557–579.

Hlushach, D., & Zhmurko, V. (2021). Vplyv tryvalosti fotoperiodu na biolohichni vlastyvosti bakteriy hrupy PGPR ryzosfery soi kulturnoyi (Glycine max (L.) Merr.) [Influence of the photoperiod duration on the biological properties of PGPR-bacteria of the soybean rhizosphere (Glycine max (L.) Merr.)]. The Journal of V. N. Karazin Kharkiv National University, Series Biology, 37, 87–94 (in Ukrainian).

Jaiswal, D. K., Verma, J. P., Prakash, S., Meena, V. S., & Meena, R. S. (2016). Potassium as an important plant nutrient in sustainable agriculture: A state of the art. In: Meena, V. S., Verma, J. P., Verma, J. K., & Meena, R. S. (Eds.). Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi. Pp. 21–29.

Kape, R., Parniske, M., & Werner, D. (1991). Chemotaxis and nod gene activity of Bradyrhizobium japonicum in response to hydroxycinnamic acids and isoflavonoids. Applied and Environmental Microbiology, 57(1), 316–319.

Kirakosyan, A., Kaufman, P., Nelson, R. L., Kasperbauer, M. J., Duke, J. A., Seymour, E., Chang, S. C., Warber, S., & Bolling, S. (2006). Isoflavone levels in five soybean (Glycine max) genotypes are altered by phytochrome-mediated light treatments. Journal of Agricultural and Food Chemistry, 54(1), 54–58.

Krutylo, D. V., & Nadkernychna, O. V. (2023). Features of local bradyrhizobia populations after long-term period in the soil without a host plant. Mikrobiolohichnyi Zhurnal, 85(5), 20–30.

Kwasny, S. M., & Opperman, T. J. (2010). Static biofilm cultures of Gram-positive pathogens grown in a microtiter format used for anti-biofilm drug discovery. Current Protocols in Pharmacology, 50(1), 13A.8.1–13A.8.23.

Lee, H. I., Lee, J. H., Park, K. H., Sangurdekar, D., & Chang, W. S. (2012). Effect of soybean coumestrol on Bradyrhizobium japonicum nodulation ability, biofilm formation, and transcriptional profile. Applied and Environmental Microbiology, 78(8), 2896–2903.

Li, Y., Hou, Z., Li, W., Li, H., Lu, S., Gan, Z., Du, H., Li, T., Zhang, Y., Kong, F., Cheng, Y., He, M., Ma, L., Liao, C., Li, Y., Dong, L., Liu, B., & Cheng, Q. (2021). The legume-specific transcription factor E1 controls leaf morphology in soybean. BMC Plant Biology, 21(1), 531.

Lin, X., Liu, B., Weller, J. L., Abe, J., & Kong, F. (2021). Molecular mechanisms for the photoperiodic regulation of flowering in soybean. Journal of Integrative Plant Biology, 63(6), 981–994.

Meena, V. S., Bahadur, I., Maurya, B. R., Kumar, A., Meena, R. K., Meena, S. K., & Verma, J. P. (2016). Potassium-solubilizing microorganism in evergreen agriculture: An overview. In: Meena, V. S., Verma, J. P., Verma, J. K., & Meena, R. S. (Eds.). Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi. Pp. 1–20.

Pettigrew, W. T. (2008). Potassium influences on yield and quality production for maize, wheat, soybean and cotton. Physiologia Plantarum, 133(4), 670–681.

Porro, M., Viti, S., Antoni, G., & Neri, P. (1981). Modifications of the Park-Johnson ferricyanide submicromethod for the assay of reducing groups in carbohydrates. Analytical Biochemistry, 118(2), 301–306.

Pramanik, M. H. R., Nagai, M., Asao, T., Matsui, Y., & Matsuyama, T. (2000). Effects of temperature and photoperiod on phytotoxic root exudates of cucumber (Cucumis sativus) in hydroponic culture. Journal of Chemical Ecology, 26(8), 1953–1967.

Qin, Y., Angelini, L. L., & Chai, Y. (2022). Bacillus subtilis cell differentiation, biofilm formation and environmental prevalence. Microorganisms, 10(6), 1108.

Raghavendra, M. P., Chandra Nayaka, S., & Nuthan, B. R. (2016). Role of rhizosphere microflora in potassium solubilization. In: Meena, V. S., Verma, J. P., Verma, J. K., & Meena, R. S. (Eds.). Potassium solubilizing microorganisms for sustainable agriculture. Springer, New Delhi. Pp. 43–59.

Rinaudi, L. V., & Giordano, W. (2010). An integrated view of biofilm formation in rhizobia. FEMS Microbiology Letters, 304(1), 1–11.

Roeber, V. M., Schmülling, T., & Cortleven, A. (2022). The photoperiod: Handling and causing stress in plants. Frontiers in Plant Science, 12, 781988.

Shah, A., Nazari, M., Antar, M., Msimbira, L. A., Naamala, J., Lyu, D., Rabileh, M., Zajonc, J., & Smith, D. L. (2021). PGPR in agriculture: A sustainable approach to increasing climate change resilience. Frontiers in Sustainable Food Systems, 5, 667546.

Stiefel, P., Rosenberg, U., Schneider, J., Mauerhofer, S., Maniura-Weber, K., & Ren, Q. (2016). Is biofilm removal properly assessed? Comparison of different quantification methods in a 96-well plate system. Applied Microbiology and Biotechnology, 100(9), 4135–4145.

Tasma, I. M., & Shoemaker, R. C. (2003). Mapping flowering time gene homologs in soybean and their association with maturity loci. Crop Science, 43(1), 319–328.

Upadhyay, S. K., Srivastava, A. K., Rajput, V. D., Chauhan, P. K., Bhojiya, A. A., Jain, D., Chaubey, G., Dwivedi, P., Sharma, B., & Minkina, T. (2022). Root exudates: Mechanistic insight of plant growth promoting rhizobacteria for sustainable crop production. Frontiers in Microbiology, 13, 916488.

Vives-Peris, V., de Ollas, C., Gómez-Cadenas, A., & Pérez-Clemente, R. M. (2020). Root exudates: From plant to rhizosphere and beyond. Plant Cell Reports, 39(1), 3–17.

Wang, L., Chen, M., Lam, P. Y., Dini-Andreote, F., Dai, L., & Wei, Z. (2022). Multifaceted roles of flavonoids mediating plant-microbe interactions. Microbiome, 10, 233.

Watt, M., & Evans, J. R. (1999). Linking development and determinacy with organic acid efflux from proteoid roots of white lupin grown with low phosphorus and ambient or elevated atmospheric CO2 concentration. Plant Physiology, 120(3), 705–716.

White, L. J., Ge, X., Brözel, V. S., & Subramanian, S. (2017). Root isoflavonoids and hairy root transformation influence key bacterial taxa in the soybean rhizosphere. Environmental Microbiology, 19(4), 1391–1406.

Yemm, E. W., & Cocking, E. C. (1955). The determination of amino-acids with ninhydrin. Analyst, 80(948), 209–213.

Zhalnina, K., Louie, K. B., Hao, Z., Mansoori, N., da Rocha, U. N., Shi, S., Cho, H., Karaoz, U., Loqué, D., Bowen, B. P., Firestone, M. K., Northen, T. R., & Brodie, E. L. (2018). Dynamic root exudate chemistry and microbial substrate preferences drive patterns in rhizosphere microbial community assembly. Nature Microbiology, 3(4), 470–480.