Dual inoculation of Bradyrhizobium and Enterobacter alleviates the adverse effect of salinity on Glycine max seedling

  • Mona S. AGHA Botany Department, Faculty of Science, Mansoura University, Mansoura, 35516 (EG)
  • Mohamed A. ABBAS Botany Department, Faculty of Science, Mansoura University, Mansoura, 35516 (EG)
  • Mahmoud R. SOFY otany and Microbiology Department, Faculty of Science, Al-Azhar University, 11884 Nasr City, Cairo (EG)
  • Samia A. HAROUN Botany Department, Faculty of Science, Mansoura University, Mansoura, 35516 (EG)
  • Amr M. MOWAFY Botany Department, Faculty of Science, Mansoura University, Mansoura, 35516;Department of Biological Sciences, Faculty of Science, New Mansoura University, New Mansoura city (EG)
Keywords: antioxidant enzymes, endophytic, osmotic pressure, soybean


The aid of beneficial microbes, which is a well-accepted strategy, may improve plant salt tolerance. However, the mechanisms that underpin it are unclear. In this study, seedling experiments were carried out to assess the effect of Bradyrhizobium and Enterobacter on the germination, growth, nonenzymatic and enzymatic content in soybean (Glycine max L.) under salt stress. Water was sprayed on the seeds as a control, and with 75 mM, 150 mM NaCl as salt stress. The findings demonstrate that salt stress (75, 150 mM) caused a significant decrease in germination, morphological criteria, and membrane stability index (MSI) when compared to control seeds but increased lipid peroxidation (MDA), electrolyte leakage (EL), osmotic pressure, proline, citric acid, sugar content, antioxidant enzymes. Furthermore, endophytic Bradyrhizobium and Enterobacter inoculation resulted in a significant rise in all of the above metrics.; however, these treatments resulted in significant reductions in ROS, EL, and MDA in stressed plants. Finally, the findings showed that combining Bradyrhizobium and Enterobacter was the most efficient in reducing the harmful effects of salt on soybean plants by boosting antioxidant up-regulation and lowering membrane leakage and ROS.


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Abu-Shahba MS, Mansour MM, Mohamed HI, Sofy MR (2021). Comparative cultivation and biochemical analysis of iceberg lettuce grown in sand soil and hydroponics with or without microbubbles and macrobubbles. Journal of Soil Science and Plant Nutrition 21(1):389-403. https://doiorg/10.1007/s42729-020-00368-x

Adams JB (1978). The inactivation and regeneration of peroxidase in relation to the high temperature–short time processing of vegetables. International Journal of Food Science & Technology 13(4):281-297. https://doi.org/10.1111/j.1365-2621.1978.tb00806.x

Adhikari B, Dhungana SK, Kim ID, Shin DH (2020). Effect of foliar application of potassium fertilizers on soybean plants under salinity stress. Journal of the Saudi Society of Agricultural Sciences 19(4):261-269. https://doi.org/10.1016/j.jssas.2019.02.001

Agarwal S, Shaheen R (2007). Stimulation of antioxidant system and lipid peroxidation by abiotic stresses in leaves of Momordica charantia. Brazilian Journal of Plant Physiology 19(2):149-161. https://doi.org/10.1590/s1677-04202007000200007

Alexieva V, Sergiev I, Mapelli S, Karanov E (2001). The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant, Cell & Environment 24(12):1337-1344. https://doi.org/10.1046/j.1365-3040.2001.00778.x

Alraey DA, Haroun SA, Omar MN, Abd-ElGawad AM, El-Shobaky AM, Mowafy AM (2019). Fluctuation of essential oil constituents in Origanum syriacum subsp. sinaicum in response to plant growth promoting bacteria. Journal of Essential Oil Bearing Plants 22(4):1022-1033. https://doi.org/10.1080/0972060X.2019.1661794

Arif Y, Singh P, Siddiqui H, Bajguz A, Hayat S (2020). Salinity induced physiological and biochemical changes in plants: An omic approach towards salt stress tolerance. Plant Physiology and Biochemistry 156:64-77. https://doi.org/10.1016/j.plaphy.2020.08.042

Assaha DV, Ueda A, Saneoka H, Al-Yahyai R, Yaish MW (2017). The role of Na+ and K+ transporters in salt stress adaptation in glycophytes. Frontiers in Physiology 8:509. https://doi.org/10.3389/fphys.2017.00509

Bates LS, Waldren RP, Teare ID (1973). Rapid determination of free proline for water-stress studies. Plant and Soil 39(1):205-207. https://doi.org/10.1007/BF00018060

Brilli F, Pollastri S, Raio A, Baraldi R, Neri L, Bartolini P, Podda A, Loreto F, Maserti BE, Balestrini R (2019). Root colonization by Pseudomonas chlororaphis primes tomato (Lycopersicum esculentum) plants for enhanced tolerance to water stress. Journal of plant physiology 232:82-93. https://doi.org/10.1016/j.jplph.2018.10.029

Cappuccino JG, Natalie S (2005). Microbiology: A Laboratory Manual.

Cen H, Wang T, Liu H, Tian D, Zhang Y (2020). Melatonin application improves salt tolerance of alfalfa (Medicago sativa L.) by enhancing antioxidant capacity. Plants 9(2):220. https://doi.org/10.3390/plants9020220.

Chung YS, Kim K-S, Hamayun M, Kim Y (2020). Silicon confers soybean resistance to salinity stress through regulation of reactive oxygen and reactive nitrogen species. Frontiers in Plant Science 10:1725. https://doi.org/10.3389/fpls.2019.01725

Desoky E-SM, Saad AM, El-Saadony MT, Merwad A-RM, Rady MM (2020). Plant growth-promoting rhizobacteria: Potential improvement in antioxidant defense system and suppression of oxidative stress for alleviating salinity stress in Triticum aestivum (L.) plants. Biocatalysis and Agricultural Biotechnology 30:101878. https://doi.org/10.1016/j.bcab.2020.101878

Egamberdieva D, Wirth S, Jabborova D, Räsänen LA, Liao H (2017). Coordination between Bradyrhizobium and Pseudomonas alleviates salt stress in soybean through altering root system architecture. Journal of Plant Interactions 12(1):100-107. https://doi.org/10.1080/17429145.2017.1294212

El-Beltagi HS, Mohamed HI, Megahed BM, Gamal M, Safwat G (2018). Evaluation of some chemical constituents, antioxidant, antibacterial and anticancer activities of Beta vulgaris L. root. Fresenius Environmental Bulletin 27(9):6369-6378.

El-Beltagi HS, Mohamed HI, Safwat G, Gamal M, Megahed B (2019). Chemical composition and biological activity of Physalis peruviana L. Gesunde Pflanzen 71:113-122.

El-Beltagi HS, Sofy MR, Aldaej MI, Mohamed HI (2020). Silicon alleviates copper toxicity in flax plants by up-regulating antioxidant defense and secondary metabolites and decreasing oxidative damage. Sustainability 12(11):4732. https://doi.org/10.3390/su12114732

Etesami H (2020). Enhanced phosphorus fertilizer use efficiency with microorganisms’ nutrient dynamics for sustainable crop production. Springer, 215-245. https://doi.org/10.1007/978-981-13-8660-2_8

Etesami H, Alikhani HA (2019). Halotolerant plant growth-promoting fungi and bacteria as an alternative strategy for improving nutrient availability to salinity-stressed crop plants saline soil-based agriculture by halotolerant microorganisms. Springer, 103-146. https://doi.org/10.1007/978-981-13-8335-9_5

Gebresenbet F, Wondemagegnehu DY (2021). New dimensions in the Grand Ethiopian Renaissance dam negotiations: ontological Security in Egypt and Ethiopia. African Security 1-27. https://doi.org/10.1080/19392206.2021.1905921

Gomez KA, Gomez AA (1984). Statistical Procedures for Agricultural Research. John Wiley & Sons Inc, Singapore.

Gordon SA, Weber RP (1951). Colorimetric estimation of indoleacetic acid. Plant Physiology 26(1):192-195. https://doi.org/10.1104/pp.26.1.192

Gupta S, Pandey S (2020). Enhanced salinity tolerance in the common bean (Phaseolus vulgaris) plants using twin ACC deaminase producing rhizobacterial inoculation. Rhizosphere 16:100241. https://doi.org/10.1016/j.rhisph.2020.100241

Harman GE, Doni F, Khadka RB, Uphoff N (2019). Endophytic strains of Trichoderma increase plants’ photosynthetic capability. Journal of Applied Microbiology 130(2):529-546. https://doi.org/10.1111/jam.14368

Hernández JA, Almansa MS (2002). Short-term effects of salt stress on antioxidant systems and leaf water relations of pea leaves. Physiologia Plantarum 115(2):251-257. https://doi.org/10.1034/j.1399-3054.2002.1150211.x

Hmaeid N, Wali M, Metoui-Ben Mahmoud O, Pueyo JJ, Ghnaya T, Abdelly C (2019). Efficient rhizobacteria promote growth and alleviate NaCl-induced stress in the plant species Sulla carnosa. Applied Soil Ecology 133:104-113. https://doi.org/10.1016/j.apsoil.2018.09.011

Holbrook AA, Edge WJW, Bailey F (1961). Spectrophotometric method for determination of gibberellic acid gibberellinS. Advances in Chemistry 28:159-167.

Ilangumaran G, Smith DL (2017). Plant growth promoting rhizobacteria in amelioration of salinity stress: a systems biology perspective. Frontiers in Plant Science 8:1768. https://doi.org/10.3389/fpls.2017.01768

Irigoyen JJ, Einerich DW, Sánchez-Díaz M (1992). Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiologia Plantarum 84(1):55-60. https://doi.org/10.1111/j.1399-3054.1992.tb08764.x

Kolthoff IM, Sandell EB (1948). Colorimetric methods of analysis. The Journal of Physical and Colloid Chemistry 52(7):1265-1266. https://doi.org/10.1021/j150463a020

Komaresofla BR, Alikhani HA, Etesami H, Khoshkholgh-Sima NA (2019). Improved growth and salinity tolerance of the halophyte Salicornia sp. by co–inoculation with endophytic and rhizosphere bacteria. Applied Soil Ecology 138:160-170. https://doi.org/10.1016/j.apsoil.2019.02.022.

Kumar A, Singh S, Gaurav AK, Srivastava S, Verma JP (2020). Plant growth-promoting bacteria: biological tools for the mitigation of salinity stress in plants. Frontiers in Microbiology 11:1216. https://doi.org/10.3389/fmicb.2020.01216

Lastochkina O (2019). Bacillus subtilis-mediated abiotic stress tolerance in plants. In: Islam MT, Rahman MM, Pandey P, Boehme MH, Haesaert G (Eds). Bacilli and Agrobiotechnology: Phytostimulation and Biocontrol: Volume 2. Springer International Publishing, Cham, pp 97-133. https://doi.org/10.1007/978-3-030-15175-1-6

Lastochkina O, Pusenkova L, Garshina D, Yuldashev R, Shpirnaya I, Kasnak C, … Aliniaeifard S (2020). The effect of endophytic bacteria Bacillus subtilis and salicylic acid on some resistance and quality traits of stored Solanum tuberosum L. tubers infected with Fusarium dry rot. Plants 9(6):738. https://doi.org/10.3390/plants9060738

Li X, Sun P, Zhang Y, Jin C, Guan C (2020). A novel PGPR strain Kocuria rhizophila Y1 enhances salt stress tolerance in maize by regulating phytohormone levels, nutrient acquisition, redox potential, ion homeostasis, photosynthetic capacity and stress-responsive genes expression. Environmental and Experimental Botany 174:104023. https://doi.org/10.1016/j.envexpbot.2020.104023

Maguire JD (1962). Speed of germination-aid in selection and evaluation for seedling emergence and vigor. Crop Science 2(2). https://doi.org/10.2135/cropsci1962.0011183X000200020033x

Mahmoud LM, Dutt M, Shalan AM, El-Kady ME, El-Boray MS, Shabana YM, Grosser JW (2020). Silicon nanoparticles mitigate oxidative stress of in vitro-derived banana (Musa acuminata ‘Grand Nain’) under simulated water deficit or salinity stress. South African Journal of Botany 132:155-163. https://doi.org/10.1016/j.sajb.2020.04.027

Mansour E, Moustafa ES, Desoky E-SM, Ali M, Yasin MA, Attia A, Alsuhaibani N, Tahir MU, El-Hendawy S (2020). Multidimensional evaluation for detecting salt tolerance of bread wheat genotypes under actual saline field growing conditions. Plants 9(10):1324. https://doi.org/10.3390/plants9101324

Matthews S, Noli E, Demir I, Khajeh-Hosseini M, Wagner M-H (2012). Evaluation of seed quality: from physiology to international standardization. Seed Science Research 22(S1):69-S73. https://doi.org/10.1017/S0960258511000365

Mbarki S, Sytar O, Cerda A, Zivcak M, Rastogi A, He X, Zoghlami A, Abdelly C, Brestic M (2018). Strategies to mitigate the salt stress effects on photosynthetic apparatus and productivity of crop plants. Salinity Responses and Tolerance in Plants 1:85-136. https://doi.org/10.1007/978-3-319-75671-4-4

Meena M, Swapnil P, Divyanshu K, Kumar S, Harish, Tripathi YN, Zehra A, Marwal A, Upadhyay RS (2020). PGPR-mediated induction of systemic resistance and physiochemical alterations in plants against the pathogens: Current perspectives. Journal of Basic Microbiology 60(10):828-861. https://doi.org/10.1002/jobm.202000370

Megahed A, El-Dougdoug K, Othman B, Lashin S, Ibrahim M, Sofy A (2012). A new Egyptian satellite strain of cucumber mosaic cucumovirus. International Journal of Virology 8(3):240-257. https://doi.org/10.3923/ijv.2012.240.257

Megahed A, KhA E-D, Othman B, Lashin S, Ibrahim M, Sofy A (2013). Induction of resistance in tomato plants against tomato mosaic tobamovirus using beneficial microbial isolates. Pakistan Journal of Biological Sciences16(8):385-390. https://doi.org/10.3923/pjbs.2013.385.390

Mishra NP, Mishra RK, Singhal GS (1993). Changes in the activities of anti-oxidant enzymes during exposure of intact wheat leaves to strong visible light at different temperatures in the presence of protein synthesis inhibitors. Plant Physiology 102(3):903-910. https://doi.org/10.1104/pp.102.3.903

Mohamed HI, Aly AA, Mansour MTM, El-Samawaty AMA (2012). Association of oxidative stress components with resistance to flax powdery mildew. Tropical Plant Pathology 37(6):386-392.

Mohamed HI, Elsherbiny EA, Abdelhamid MT (2016). Physiological and biochemical responses of Vicia faba Plants to foliar application of zinc and iron. Gesunde Pflanzen 68(4):201-212.

Mohamed HI, El-Beltagi HS, Aly AA, Latif HH (2018). The role of systemic and non systemic fungicides on the physiological and biochemical parameters in Gossypium hirsutum plant, implications for defense responses. Fresenius Environmental Bulletin 7(12):8585-8593.

Mowafy AM, Fawzy MM, Gebreil A, Elsayed A (2021). Endophytic Bacillus, Enterobacter, and Klebsiella enhance the growth and yield of maize. Acta Agriculturae Scandinavica, Section B - Soil & Plant Science 71(4):237-246. https://doi.org/10.1080/09064710.2021.1880621

Nawaz A, Shahbaz M, Imran A, Marghoob MU, Imtiaz M, Mubeen F (2020). Potential of salt tolerant PGPR in growth and yield augmentation of wheat (Triticum aestivum L.) under saline conditions. Frontiers in Microbiology 11:2019. https://doi.org/10.3389/fmicb.2020.02019

Ngalimat MS, Mohd Hata E, Zulperi D, Ismail SI, Ismail MR, Mohd Zainudin NAI, Saidi NB, Yusof MT (2021). Plant growth-promoting bacteria as an emerging tool to manage bacterial rice pathogens. Microorganisms 9(4):682. https://doi.org/10.3390/microorganisms9040682

Oktay M, Küfreviolu I, Kocaçalişkan I, Şaklrolu H (1995). Polyphenoloxidase from Amasya apple. Journal of Food Science 60(3):494-496. https://doi.org/10.1111/j.1365-2621.1995.tb09810.x

Rajendra L, Samiyappan R, Raguchander TG, Saravanakumar D (2006). Endophytic bacterial induction of defence enzymes against bacterial blight of cotton. Phytopathologia Mediterranea 45(3):203-214.

Reyhaneh A, Hassan F, Mohammad KH (2013). Can bulk and nanosized titanium dioxide particles improve seed germination features of wheatgrass (Agropyron desertorum). Notulae Scientia Biologicae 5(3). https://doi.org/10.15835/nsb539072

Sairam RK (1994). Effect of moisture stress on physiological activities of two contrasting wheat genotypes. Indian Journal of Experimental Biology 3:584-593.

Sandrasagaran UM, Subramaniam S, Murugaiyah V (2014). New perspective of Dendrobium crumenatum orchid for antimicrobial activity against selected pathogenic bacteria. Pakistan Journal of Botany 46(2):719-724.

Schwyn B, Neilands JB (1987). Universal chemical assay for the detection and determination of siderophores. Analytical Biochemistry 160(1):47-56. https://doi.org/10.1016/0003-2697(87)90612-9

Sharaf AEM, Farghal II, Sofy MR (2009). Response of broad bean and lupin plants to foliar treatment with boron and zinc. Australian Journal of Basic and Applied Sciences 3:2226-2231.

Silva PCC, Azevedo Neto ADd, Gheyi HR, Ribas RF, Silva CRdR, Cova AMW (2020). Salt tolerance induced by hydrogen peroxide priming on seed is related to improvement of ion homeostasis and antioxidative defense in sunflower plants. Journal of Plant Nutrition 44(8):1207-1221. https://doi.org/10.1080/01904167.2020.1862202

Sofy AR, Sofy MR, Hmed AA, Dawoud RA, Alnaggar AE-AM, Soliman AM, El-Dougdoug NK (2021a). Ameliorating the adverse effects of tomato mosaic tobamovirus infecting tomato plants in Egypt by boosting immunity in tomato plants using zinc oxide nanoparticles. Molecules 26(5):1337. https://doi.org/10.3390/molecules26051337

Sofy AR, Sofy MR, Hmed AA, Dawoud RA, Refaey EE, Mohamed HI, El-Dougdoug NK (2021b). Molecular characterization of the Alfalfa mosaic virus infecting Solanum melongena in Egypt and the control of its deleterious effects with melatonin and salicylic acid. Plants 10(3):459. https://doi.org/10.3390/plants10030459

Sofy M, Mohamed H, Dawood M, Abu-Elsaoud A, Soliman M (2021c). Integrated usage of arbuscular mycorrhizal and biochar to ameliorate salt stress on spinach plants. Archives of Agronomy and Soil Science 1-22. https://doi.org/10.1080/03650340.2021.1949709

Sofy MR, Aboseidah AA, Heneidak SA, Ahmed HR (2021d). ACC deaminase containing endophytic bacteria ameliorate salt stress in Pisum sativum through reduced oxidative damage and induction of antioxidative defense systems. Environmental Science and Pollution Research 1-21. https://doi.org/10.1007/s11356-021-13585-3

Sofy MR, Elhawat N, Alshaal T (2020a). Glycine betaine counters salinity stress by maintaining high K+/Na+ ratio and antioxidant defense via limiting Na+ uptake in common bean (Phaseolus vulgaris L.). Ecotoxicology and Environmental Safety 200:110732. https://doi.org/10.1016/j.ecoenv.2020.110732

Sofy MR, Elhindi KM, Farouk S, Alotaibi MA (2020b). Zinc and paclobutrazol mediated regulation of growth, upregulating antioxidant aptitude and plant productivity of pea plants under salinity. Plants 9(9):1197. https://doi.org/10.3390/plants9091197

Sofy AR, Dawoud RA, Sofy MR, Mohamed HI, Hmed AA, El-Dougdoug NK (2020c). Improving regulation of enzymatic and nonenzymatic antioxidants and stress-related gene stimulation in Cucumber mosaic cucumovirus infected cucumber plants treated with glycine betaine, chitosan and combination. Molecules 25:2341. https://doi.org/10.3390/molecules25102341

Szalai G, Janda T, Páldi E, Szigeti Z (1996). Role of light in the development of post-chilling symptoms in maize. Journal of Plant Physiology 148(3):378-383. https://doi.org/10.1016/S0176-1617(96)80269-0

Valenzuela-Aragon B, Parra-Cota FI, Santoyo G, Arellano-Wattenbarger GL, de los Santos-Villalobos S (2019). Plant-assisted selection: a promising alternative for in vivo identification of wheat (Triticum turgidum L. subsp. Durum) growth promoting bacteria. Plant and Soil 435(1):367-384. https://doi.org/10.1007/s11104-018-03901-1

Vashisth A, Nagarajan S (2010). Effect on germination and early growth characteristics in sunflower (Helianthus annuus) seeds exposed to static magnetic field. Journal of Plant Physiology 167(2):149-156. https://doi.org/10.1016/j.jplph.2009.08.011

Wang M, Ding F, Zhang S (2020). Mutation of SlSBPASE aggravates chilling-induced oxidative stress by impairing glutathione biosynthesis and suppressing ascorbate-glutathione recycling in tomato plants. Frontiers in Plant Science 11:2135. https://doi.org/10.3389/fpls.2020.565701

Yadav AN, Verma P, Sachan SG, Kaushik R, Saxena AK (2018). Psychrotrophic microbiomes: molecular diversity and beneficial role in plant growth promotion and soil health. Microorganisms for green revolution. Springer, pp 197-240. https://doi.org/10.1007/978-981-10-7146-1-11

Yasmin H, Naeem S, Bakhtawar M, Jabeen Z, Nosheen A, Naz R, Keyani R, Mumtaz S, Hassan MN (2020). Halotolerant rhizobacteria Pseudomonas pseudoalcaligenes and Bacillus subtilis mediate systemic tolerance in hydroponically grown soybean (Glycine max L.) against salinity stress. PLoS One 15(4):e0231348. https://doi.org/10.1371/journal.pone.0231348

Zhang F, Xu X, Huo Y, Xiao Y (2019). Trichoderma-inoculation and mowing synergistically altered soil available nutrients, rhizosphere chemical compounds and soil microbial community, potentially driving alfalfa growth. Frontiers in Microbiology 9. https://doi.org/10.3389/fmicb.2018.03241

Zhang Y, Li Y, Hassan MJ, Li Z, Peng Y (2020). Indole-3-acetic acid improves drought tolerance of white clover via activating auxin, abscisic acid and jasmonic acid related genes and inhibiting senescence genes. BMC Plant Biology 20(1):1-12. https://doi.org/10.1186/s12870-020-02354-y

How to Cite
AGHA, M. S., ABBAS, M. A., SOFY, M. R., HAROUN, S. A., & MOWAFY, A. M. (2021). Dual inoculation of Bradyrhizobium and Enterobacter alleviates the adverse effect of salinity on Glycine max seedling. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 49(3), 12461. https://doi.org/10.15835/nbha49312461
Research Articles
DOI: 10.15835/nbha49312461