Beneficial traits of grain-residing endophytic communities in wheat with different sensitivity to Pseudomonas syringae

  • A. Pastoshchuk Taras Shevchenko National University of Kyiv
  • Y. Yumyna Taras Shevchenko National University of Kyiv
  • P. Zelena Taras Shevchenko National University of Kyiv
  • V. Nudha Institute of Microbiology Diagnostics
  • V. Yanovska Institute of Microbiology Diagnostics
  • M. Kovalenko Taras Shevchenko National University of Kyiv
  • N. Taran Taras Shevchenko National University of Kyiv
  • V. Patyka D. K. Zabolotny Institute of Microbiology and Virology of National Academy of Sciences of Ukraine
  • L. Skivka Taras Shevchenko National University of Kyiv
Keywords: basal glume rot of wheat; leaf blight; agrobiotechnology; Bacillus; Paenibacillus; Brevibacillus; wheat grain-residing endophyte.


Deep insight into compositional and functional features of endophytic bacterial communities residing in wheat grains opens the way to the use of their plant growth promoting and biocontrol abilities in agricultural biotechnology. The aim of this work was to compare grain-residing endophytes from winter wheat varieties with different sensitivity to Pseudomonas syringae pv. atrofaciens (McCulloch) and to examine their plant-beneficial traits and antagonistic effects. Grain-residing bacteria were isolated from surface-sterilized grains of three wheat varieties sown in Ukraine following a culture-dependent protocol, and were screened for their plant growth promotion (PGP) and antagonistic properties. Bacterial morphotypes were represented by gram-negative rods, endospore-forming bacilli and gram-positive cocci. Different resistance to phytopathogenic pseudomonads was associated with distinctive quantitative and functional features of grain-residing endophytic communities. High resistance to P. syringae was coupled with the prevalence of gram-negative rods in the endophytic community, the highest proportion of endophytic bacteria possessing three PGP activities (phosphate solubilization, nitrogen fixation and production of indolic compounds) simultaneously, and with the most potent antagonistic activity of grain-residing endospore-forming bacilli. In total, five grain-residing isolates, which were obtained from three wheat varieties (two isolates from varieties with medium and high resistance and one – from a low-resistant variety), demonstrated ability to restrain P. syringae pv. atrofaciens (McCulloch) growth. Two isolates (P6 and P10) which were obtained from the high-resistant wheat variety Podolyanka and were assigned to Paenibacillus and Brevibacillus genera according to their biochemical profiling and MS-DS identification, showed the most potent antagonistic effects as indicated by maximum inhibition zone in agar well diffusion assay. These results shed light on the association of the features of grain-residing endophytic bacteria with wheat resistance to phytopathogenic pseudomonads. Isolates from the high-resistant wheat variety can be recommended for grain dressing as plant growth promoting and biocontrol agents for P. syringae pv. atrofaciens (McCulloch).


Aebi, H. (1984). Catalase in vitro. Methods Enzymology, 105, 121–126.

Afzal, I., Shinwari, Z. K., Sikandar, S., & Shahzad, S. (2019). Plant beneficial endophytic bacteria: Mechanisms, diversity, host range and genetic determinants. Microbiological Research, 221, 36–49.

Agarwal, H., Dowarah, B., Baruah, P. M., Bordoloi, K. S., Krishnatreya, D. B., & Agarwala, N. (2020). Endophytes from Gnetum gnemon L. can protect seedlings against the infection of phytopathogenic bacterium Ralstonia solanacearum as well as promote plant growth in tomato. Microbiological Research, 238, 126503.

Alikhani, H., Saleh-Rastin, N., & Antoun, H. (2006). Phosphate solubilization activity of rhizobia native to Iranian soils. Plant and Soil, 287, 35.

Bates, L. S., Waldren, R. P., & Teare, I. D. (1973). Rapid determination of free proline for water-stress studies. Plant and Soil, 39, 205–207.

Beyer, W. F., & Fridovich, I. (1987). Assaying for superoxide dismutase activity: Some large consequences of minor changes in conditions. Analytical Biochemistry, 161, 559–566.

Bezpal’ko, V. V., Stankevych, S. V., Zhukova, L. V., Zabrodina, I. V., Turenko, V. P., Horyainova, V. V., Poedinceva, А. A., Batova, O. M., Zayarna, O. Y., Bondarenko, S. V., Dolya, M. M., Mamchur, R. M., Drozd, P. Y., Sakhnenko, V. V., & Matsyura, A. V. (2020). Pre-sowing seed treatment in winter wheat and spring barley cultivation. Ukrainian Journal of Ecology, 10(6), 255–268.

Compant, S., Cambon, M. C., Vacher, C., Mitter, B., Samad, A., & Sessitsch, A. (2020). The plant endosphere world – bacterial life within plants. Environmental Microbiology, 23, 1812–1829.

Eevers, N., Gielen, M., Sánchez-López, A., Jaspers, S., White, J. C., Vangronsveld, J., & Weyens, N. (2015). Optimization of isolation and cultivation of bacterial endophytes through addition of plant extract to nutrient media. Microbial Biotechnology, 8(4), 707–715.

Eid, A. M., Fouda, A., Abdel-Rahman, M. A., Salem, S. S., Elsaied, A., Oelmüller, R., Hijri, M., Bhowmik, A., Elkelish, A., & Hassan, S. E.-D. (2021). Harnessing bacterial endophytes for promotion of plant growth and biotechnological applications: An overview. Plants, 10(5), 935.

Geisen, S., Kostenko, O., Cnossen, M. C., Ten Hooven, F. C., Vreš, B., & van der Putten, W. H. (2017). Seed and root endophytic fungi in a range expanding and a related plant species. Frontiers in Microbiology, 8, 1645.

Herrera, D., Grossi, C., Zawoznik, M., & Groppa, M. D. (2016). Wheat seeds harbour bacterial endophytes with potential as plant growth promoters and biocontrol agents of Fusarium graminearum. Microbiological Research, 186, 37–43.

Imlay, J. A. (2008). Cellular defenses against superoxide and hydrogen peroxide. Annual Review of Biochemistry, 77, 755–776.

Karthikeyan, G., Rajendran, L., Sendhilvel, V., Prabakar, K., & Raguchander, T. (2021). Diversity and functions of secondary metabolites secreted by epi-endophytic microbes and their interaction with phytopathogens. In: Sudisha, J. (Ed.). Biocontrol agents and secondary metabolites: Applications and immunization for plant growth and protection. Woodhead Publishing, Cambridge. Pp. 495–517.

Kazempour, M., Kheyrgoo, M., Pedramfar, H., & Rahimian, H. (2010). Isolation and identification of bacterial glum blotch and leaf blight on wheat (Triticum aestivum L.) in Iran. African Journal of Biotechnology, 9, 2860–2865.

Kumar, G., & Knowles, N. R. (1993). Changes in lipid peroxidation and lipolytic and free-radical scavenging enzyme activities during aging and sprouting of potato (Solanum tuberosum) seed-tubers. Plant Physiology, 102(1), 115–124.

Kuźniar, A., Włodarczyk, K., Grządziel, J., Woźniak, M., Furtak, K., Gałązka, A., Dziadczyk, E., Skórzyńska-Polit, E., & Wolińska, A. (2020). New insight into the composition of wheat seed microbiota. International Journal of Molecular Sciences, 21(13), 4634.

Lamichhane, J. R., Messean, A., & Morris, C. E. (2015). Insights into epidemiology and control of diseases of annual plants caused by the Pseudomonas syringae species complex. Journal of General Plant Pathology, 81, 331–350.

Maheshwari, D. K. (2017). Endophytes: Biology and biotechnology. Springer International Publishing AG. Cambridge.

Makar, O., Kuźniar, A., Patsula, O., Kavulych, Y., Kozlovskyy, V., Wolińska, A., Skórzyńska-Polit, E., Vatamaniuk, O., Terek, O., & Romanyuk, N. (2021). Bacterial endophytes of spring wheat grains and the potential to acquire Fe, Cu, and Zn under their low soil bioavailability. Biology, 10(5), 409.

Meena, M., Divyanshu, K., Kumar, S., Swapnil, P., Zehra, A., Shukla, V., Yadav, M., & Upadhyay, R. S. (2019). Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions. Heliyon, 5(12), e02952.

Mishra, A., Singh, S. P., Mahfooz, S., Singh, S. P., Bhattacharya, A., Mishra, N., & Nautiyal, C. S. (2018). Endophyte-mediated modulation of defense-related genes and systemic resistance in Withania somnifera (L.) Dunal under Alternaria alternata stress. Applied and Environmental Microbiology, 84(8), e02845–17.

Morales-Cedeño, L. R., Orozco-Mosqueda, M. D. C., Loeza-Lara, P. D., Parra-Cota, F. I., de Los Santos-Villalobos, S., & Santoyo, G. (2020). Plant growth-promoting bacterial endophytes as biocontrol agents of pre- and post-harvest diseases: Fundamentals, methods of application and future perspectives. Microbiological Research, 242, 126612.

Mougou, I., & Boughalleb-M’hamdi, N. (2018). Biocontrol of Pseudomonas syringae pv. syringae affecting citrus orchards in Tunisia by using indigenous Bacillus spp. and garlic extract. Egyptian Journal of Biological Pest Control, 28, 60.

Muthukumar, A., Regunathan, U., & Ramasamy, N. (2017). Role of bacterial endophytes in plant disease control. In: Maheshwari, D. K., & Annapurna, K. (Eds.). Endophytes: Crop productivity and protection. Vol. 2. Springer International Publishing AG. Cham. Pp. 131–161.

Nikolić, I., Berić, T., Dimkić, I., Popović, T., Lozo, J., Fira, D., & Stanković, S. (2019). Biological control of Pseudomonas syringae pv. aptata on sugar beet with Bacillus pumilus SS-10.7 and Bacillus amyloliquefaciens (SS-12.6 and SS-38.4) strains. Journal of Applied Microbiology, 126(1), 165–176.

Papik, J., Folkmanova, M., Polivkova-Majorova, M., Suman, J., & Uhlik, O. (2020). The invisible life inside plants: Deciphering the riddles of endophytic bacterial diversity. Biotechnology Advances, 44, 107614.

Pasichnyk, L. A. (2016). The spectrum of weed phytopathogens in wheat agrophytocenosis. Mikrobiolohichnyi Zhurnal, 78(6), 19–28.

Pastoshchuk, A. Y., Skivka, L. M., Butsenko, L. M., & Patyka, V. P. (2018). Vplyv zbudnyka bazalnoho bakteriozu na prorostannia zeren ta rist parostkiv pshenytsi riznykh sortiv [Effect of causal agent of basal bacteriosis on seed germination and root growth of different wheat varieties]. Mikrobiolohiia i Biotekhnolohiia, 42, 39–48 (in Ukrainian).

Rana, A., Kabi, S. R., Verma, S., Adak, A., Pal, M., Shivay, Y. S., Prasanna, R., & Nain, L. (2015). Prospecting plant growth promoting bacteria and cyanobacteria as options for enrichment of macro- and micronutrients in grains in rice–wheat cropping sequence. Cogent Food and Agriculture, 1, 1037379.

Ridout, M. E., Schroeder, K. L., Hunter, S. S., Styer, J., & Newcombe, G. (2019). Priority effects of wheat seed endophytes on a rhizosphere symbiosis. Symbiosis, 78, 19–31.

Rychert, J., Burnham, C. A., Bythrow, M., Garner, O. B., Ginocchio, C. C., Jennemann, R., Lewinski, M. A., Manji, R., Mochon, A. B., Procop, G. W., Richter, S. S., Sercia, L., Westblade, L. F., Ferraro, M. J., & Branda, J. A. (2013). Multicenter evaluation of the Vitek MS matrix-assisted laser desorption ionization-time of flight mass spectrometry system for identification of gram-positive aerobic bacteria. Journal of Clininical Microbiology, 51(7), 2225–2231.

Sadaf, S., Nuzhat, A., & Khan, N. S. (2009). Indole acetic acid production and enhanced plant growth promotion by indigenous PSBs. African Journal of Agricultural Research, 4(11), 1312–1316.

Smirnov, O., Karpets, L. A., Zinchenko, A., Kovalenko, M., Belava, V., & Taran, N. (2020). Changes of morphofunctional traits of Triticum aestivum and Triticum dicoccum seedlings caused by polyethylene glycol-modeling drought. Journal of Central European Agriculture, 21(2), 268–274.

Tamosiune, I., Baniulis, D., & Stanys, V. (2017). Role of endophytic bacteria in stress tolerance of agricultural plants: Diversity of microorganisms and molecular mechanisms. In: Kumar, V., Kumar, M., Sharma, S., & Prasad, R. (Eds.). Probiotics in agroecosystem. Springer, Singapore. Pp. 1–29.

Umesha, S. (2020). Diversity of seed-borne bacterial phytopathogens. In: Kumar, R., & Gupta, A. (Eds.). Seed-borne diseases of agricultural crops: Detection, diagnosis and management. Springer, Singapore. Pp. 307–328.

Valencia-Botín, A. J., & Cisneros-López, M. E. (2012). A review of the studies and interactions of Pseudomonas syringae pathovars on wheat. International Journal of Agronomy, 2012, 692350.

Wang, Q., Ge, C., Xu, S. Wu, Y., Sahito, Z. A., Ma, L., Pan, F., Zhou, Q., Huang, L., Feng, Y., & Yang, X. (2020). The endophytic bacterium Sphingomonas SaMR12 alleviates Cd stress in oilseed rape through regulation of the GSH-AsA cycle and antioxidative enzymes. BMC Plant Biology, 20(1), 63.

Woźniak, M., Gałązka, A., Tyśkiewicz, R., & Jaroszuk-Ściseł, J. (2019). Endophytic bacteria potentially promote plant growth by synthesizing different metabolites and their phenotypic/physiological profiles in the Biolog GEN III MicroPlateTM test. International Journal of Molecular Sciences, 20(21), 5283.

Zeier, J. (2013). New insights into the regulation of plant immunity by amino acid metabolic pathways. Plant Cell and Environment, 36(12), 2085–103.

How to Cite
Pastoshchuk, A., Yumyna, Y., Zelena, P., Nudha, V., Yanovska, V., Kovalenko, M., Taran, N., Patyka, V., & Skivka, L. (2021). Beneficial traits of grain-residing endophytic communities in wheat with different sensitivity to Pseudomonas syringae . Regulatory Mechanisms in Biosystems, 12(3), 498-505.