ISOLATION AND CHARACTERIZATION OF MESORHIZOBIUM STRAINS FROM WILD AND CULTIVATED CHICKPEA NODULES IN THE KURDISTAN-IRAQ
DOI:
https://doi.org/10.36103/mvm17v91Keywords:
Mesorhizobium, Chickpea nodule, symbiosis, gram-negative bacteria, salinity tolerance, indigenous rhizobiaAbstract
The study was conducted to investigate symbiosis and node-forming ability of Mesorhizobium bacteria in chickpea (Cicer arietinum L.) growth under challenging environmental conditions. Nodules were collected from wild and cultivated chickpeas in the Kurdistan Region. Endophytic bacteria associated with their roots were isolated and characterized. All the 30 isolates, were found to be gram-negative with smooth, circular colonies. Most isolates (80%) were mucoid, while 20% were not and produced green colonies on bromothymol blue. Half of the isolates 50%) grew best in alkaline pH (6.6-10.5), 26.6% thrived in acidic pH (3.5-5.5), and 20% preferred neutral pH. Regarding salt tolerance, 63% of the isolates grew well in 0-4% NaCl, while 6.67% tolerated up to 6% NaCl. In terms of temperature, 56.67% grew better in the range of 15-35°C, while 36.67% thrived in 25-40°C. Some isolates had slow growth in alkaline conditions, producing blue colonies, while fast-growing isolates in acidic conditions yielded yellow colonies. This research provides insights into the adaptability of these bacteria under harsh conditions of high pH, salinity, and extreme temperatures. The outcomes offering a pathway to improving chickpea resilience and productivity in marginal lands of the Kurdistan Region and similar sites worldwide, through utilizing indigenous candidate rhizobia for adverse environments.
Received: 16/6/2024
Accepted: 23/10/2024
Published: 2026/4/30
References
Abdullaeva, Y., Ratering, S., Ambika Manirajan, B., Rosado-Porto, D., Schnell, S. and Cardinale, M. 2022. Domestication impacts the wheat-associated microbiota and the rhizosphere colonization by seed-and soil-originated microbiomes, across different fields. Frontiers in plant science, 12, 806915. https://doi.org/10.3389/fpls.2021.806915.
Adeleke, B.S., Babalola, O.O. and Glick, B.R. 2021. Plant growth-promoting root-colonizing bacterial endophytes. Rhizosphere, 20, 100433. https://doi.org/10.1016/j.rhisph.2021.100433.
Ali, S.R., Mahadiuzzaman, A., Hossain, R., Nusrin, S. and Yasmin, S. 2019. Morphological and physiological characterization of nitrogen-fixing rhizobia isolated from country bean (Lablaba perpureus) of Narail. Bangladesh. Journal of Bioscience and Biotechnology Discovery, 4 (4), 60-68. https://doi.org/10.31248/JBBD2019.100
Benjelloun, I., Thami Alami, I., Douira, A. and Udupa, S.M. 2019. Phenotypic and genotypic diversity among symbiotic and non-symbiotic bacteria present in chickpea nodules in Morocco. Frontiers in microbiology, 10, 1885. https://doi.org/10.3389/fmicb.2019.01885
Bier, R.L., Daniels, M., Oviedo-Vargas, D., Peipoch, M., Price, J.R., Omondi, E. et al. 2024. Agricultural soil microbiomes differentiate in soil profiles with fertility source, tillage, and cover crops. Agriculture, Ecosystems & Environment, 368, 109002. https://doi.org/10.1016/j.agee.2024.109002
Brasca, R., Kelterer, A.-M., Maeder, M., Alcaráz, M.R. and Culzoni, M.J. 2018. Quantum chemical computation-based strategy for alternating least squares initialization in multivariate curve resolution analysis of spectral-pH data. Microchemical Journal, 140, 183-188. https://doi.org/10.1016/j.microc.2018.04.022
Clúa, J., Roda, C., Zanetti, M.E. and Blanco, F.A. 2018. Compatibility between legumes and rhizobia for the establishment of a successful nitrogen-fixing symbiosis. Genes, 9 (3), 125. https://doi.org/10.3390/genes9030125
Coba de la Pena, T. and Pueyo, J.J. 2012. Legumes in the reclamation of marginal soils, from cultivar and inoculant selection to transgenic approaches. Agronomy for Sustainable Development, 32, 65-91. https://doi.org/10.1007/s13593-011-0024-2
Cordova-Rodriguez, A., Rentería-Martínez, M., López-Miranda, C., Guzmán-Ortíz, J. and Moreno-Salazar, S. 2022. Simple and sensitive spectrophotometric method for estimating the nitrogen-fixing capacity of bacterial cultures. MethodsX, 9, 101917. https://doi.org/10.1016/j.mex.2022.101917
Demissie, S., Natea, G. and Raga, D. 2018. Microbial quality and safety of bread sold in cafeteria, tea and bread shop of Jimma town, Oromia regional state, southwest Ethiopia. International Journal of Advanced Research, 6 (1), 771-779.
https://doi: 10.4172/2572-4134.1000122
Emitaro, W.O., Kawaka, F., Musyimi, D.M. and Adienge, A. 2024. Diversity of endophytic bacteria isolated from leguminous agroforestry trees in western Kenya. AMB Express, 14 (1), 18.
https://doi.org/10.1186/s13568-024-01676-6
Forti, L., Pezzotta, A., Zebari, M. and Zerboni, A. 2023. Geomorphology of the Central Kurdistan Region of Iraq: landscapes of the Erbil Plain between the Great Zab and Little Zab Rivers. Journal of Maps, 19 (1), 2164527. https://doi.org/10.1080/17445647.2022.2164527.
Gebremedhin, W., Assefa, F., Thuita, M.N. and Masso, C. 2018. Nutritionally versatile, abiotic stress resistant and symbiotically effective chickpea (Cicer arietinum L.) root nodulating rhizobial isolates from eastern, southeastern and southern Ethiopia. Electronic Journal of Biology.
Giongo, A., Beneduzi, A., Ambrosini, A., Vargas, L.K., Stroschein, M.R., Eltz, F.L. 2010. Isolation and characterization of two plant growth-promoting bacteria from the rhizoplane of a legume (Lupinus albescens) in sandy soil. Revista Brasileira de Ciência do Solo, 34, 361-369. https://doi.org/10.1590/S0100-06832010000200009
Goyal, R.K., Schmidt, M.A. and Hynes, M.F. 2021. Molecular biology in the improvement of biological nitrogen fixation by rhizobia and extending the scope to cereals. Microorganisms, 9 (1), 125. https://doi.org/10.3390/microorganisms9010125.
Granada Agudelo, M., Ruiz, B., Capela, D. and Remigi, P. 2023. The role of microbial interactions on rhizobial fitness. Frontiers in Plant Science, 14, 1277262. https://doi.org/10.3389/fpls.2023.1277262
Habib-ur-Rahman M, Ahmad A, Raza A, Hasnain MU, Alharby HF, Alzahrani YM, Bamagoos AA, Hakeem KR, Ahmad S, Nasim W, Ali S, Mansour F and EL Sabagh A 2022. Impact of climate change on agricultural production; Issues, challenges, and opportunities in Asia. Frontiers in Plant Science, 13, 925548. https://doi.org/10.3389/fpls.2022.925548
Hamza, T.A. and Alebejo, A.L. 2017. Isolation and characterization of rhizobia from rhizospher and root nodule of cowpea, elephant and lab plants. Int. J. Nov. Res. Interdiscip. Stud, 4, 1-7. Vol. 4, Issue 4, pp: (1-7), Month: July – August 2017, Available at: www.noveltyjournals.com
Igolkina, A.A., Noujdina, N.V., Vishnyakova, M., Longcore, T., von Wettberg, E., Nuzhdin, S.V. et al. 2023. Historical routes for diversification of domesticated chickpea inferred from landrace genomics. Molecular Biology and Evolution, 40 (6), msad110. https://doi.org/10.1093/molbev/msad110
Jadhav, K., Chavan, A., More, S., Kulkarni, S. and Karande, R. 2021. To study the seasonal incidence of gram pod borer (Helicoverpa armigera Hubner) in chickpea (Cicer arietinum L.). Journal of Pharmacognosy and Phytochemistry, 10 (6), 186-189.
doi: https://dx.doi.org/10.22271/phyto
Jain, D., Kumari, A., Saheewala, H., Sanadhya, S., Maheshwari, D., Meena, R.H. 2020. Biochemical, functional and molecular characterization of pigeon pea rhizobia isolated from semi-arid regions of India. Archives of Microbiology, 202, 1809-1816. https://doi.org/10.1007/s00203-020-01904-0
Kenasa, G., Jida, M. and Assefa, F. 2014. Characterization of phosphate solubilizing faba bean (Vicia faba L.) nodulating rhizobia isolated from acidic soils of Wollega, Ethiopia. Science, Technology and Arts Research Journal, 3 (3), 11-17.
https://doi 10.4314/star.v3i3.2
Khdir SA, Ahmad N, Hama-Ali E, Abdullah SM .2023. Genetic Diversity and Population Structure of Common Bean Genotypes Using Morphological Traits and SSR. Iraqi Journal of Agricultural Sciences 54:792-805. https://doi.org/10.36103/ijas.v54i3.1762
Kumar, A., Singh, R., Yadav, A., Giri, D., Singh, P. and Pandey, K.D. 2016. Isolation and characterization of bacterial endophytes of Curcuma longa L. 3 Biotech, 6, 1-8. Doi
L’taief, B., Sifi, B., Zaman-Allah, M., Drevon, J.-J. and Lachaâl, M. 2007. Effect of salinity on root-nodule conductance to the oxygen diffusion in the Cicer arietinum–Mesorhizobium ciceri symbiosis. Journal of plant physiology, 164 (8), 1028-1036. https://doi.org/10.1016/j.jplph.2006.05.016
Lacerda-Júnior, G.V., Noronha, M.F., Cabral, L., Delforno, T.P., De Sousa, S.T.P., Fernandes-Júnior, P.I. 2019. Land use and seasonal effects on the soil microbiome of a Brazilian dry forest. Frontiers in microbiology, 10, 648.https://doi.org/10.3389/fmicb.2019.00648
Laurette, N.N., Maxémilienne, N.B., Henri, F., Souleymanou, A., Kamdem, K., Albert, N. et al. 2015 Isolation and screening of indigenous Bambara groundnut (Vigna subterranea) nodulating bacteria for their tolerance to some environmental stresses. American Journal of Microbiological Research, 3 (2), 65-75. https://doi:10.12691/ajmr-3-2-5 58. Somasegaran, P. and Hoben, H.J. 2012. Handbook for rhizobia: methods in legume-Rhizobium technology. Springer Science & Business Media. https://doi: 10.1007/978-4613-8375-8
Maataln gn vylah, J., Sanjuan, J. and Lluch, C. 2002. Phenotypic characterization of rhizobia isolated from chickpea (Cicer arietinum) growing in Moroccan soils. Agronomie, 22 (3), 321-329.
https://DOI: 10.1051/agro:2002013
Mabrouk, Y., Hemissi, I., Salem, I.B., Mejri, S., Saidi, M. and Belhadj, O. 2018 Potential of rhizobia in improving nitrogen fixation and yields of legumes. Symbiosis, 107 (73495), 1-16. https://dx.doi.org/10.5772/intechopen.73495.
Maesen, L.v.d., Maxted, N., Javadi, F., Coles, S. and Davies, A. 2007. Taxonomy of the genus Cicer revisited. In Chickpea breeding and management, CABI Wallingford UK, pp. 14-46. https://doi.org/10.1079/9781845932138.002.
Mendoza-Suárez, M., Andersen, S.U., Poole, P.S. and Sánchez-Cañizares, C. 2021. Competition, nodule occupancy, and persistence of inoculant strains: key factors in the rhizobium-legume symbioses. Frontiers in plant science, 12, 690567. https://doi.org/10.3389/fpls.2021.690567
Muleta, A., Tesfaye, K., Haile Selassie, T.H., Cook, D.R. and Assefa, F. 2021. Phosphate solubilization and multiple plant growth promoting properties of Mesorhizobium species nodulating chickpea from acidic soils of Ethiopia. Archives of Microbiology, 203, 2129-2137.
https://doi.org/10.1007/s00203-021-02189-7
Nabintu, N.B., Ndemo, O.R., Sharwasi, N.L., Gustave, M.N., Esther, M.R. and Okoth, K.S. 2019. Indigenous rhizobia strains: The silver bullet for enhanced biological nitrogen fixation and soybean (Glycine max (L.) Merr.) yield under different soil conditions in South Kivu province, Democratic Republic of Congo. African Journal of Agricultural Research, 14 (35), 2038-2047.
https://doi.org/10.5897/AJAR2019.14457
Nath Bhowmik, S. and Das, A. 2018 Biofertilizers: a sustainable approach for pulse production. Legumes for soil health and sustainable management, 445-485.doi https://doi.org/10.1007/978-981-13-0253-4_14
Negacz, K., Malek, Ž., de Vos, A. and Vellinga, P. 2022. Saline soils worldwide: Identifying the most promising areas for saline agriculture. Journal of arid environments, 203, 104775. https://doi.org/10.1016/j.jaridenv.2022.104775
Nyaombo, J.J. 2022. Pulses Farming; An Adaptive Strategy to Climate Change in Arid and Semi-Arid Regions: A Case Study of Itigi District in Singida Region, Tanzania. In The Nature, Causes, Effects and Mitigation of Climate Change on the Environment, IntechOpen. https://doi.org/10.5772/intechopen.94777
Ondieki, D.K., Nyaboga, E.N., Wagacha, J.M. and Mwaura, F.B. 2017. Morphological and genetic diversity of Rhizobia nodulating cowpea (Vigna unguiculata L.) from agricultural soils of lower eastern Kenya. International Journal of Microbiology, 2017 (1), 8684921. https://doi.org/10.1155/2017/8684921
Rahman, M., Khatun, S., Ali, S., Yasmin, S., Kamruzzaman, M. and Rashid, M. 2018. Morpho-physiological diversity of root nodule rhizobia from mimosa (Mimosa pudica L.) and water mimosa (Neptunia oleracea L.). J Bacteriol Mycol, 5 (1), 1061. | https://www.austinpublishinggroup.com
Rai, R., Dash, P.K., Mohapatra, T. and Singh, A. 2012. Phenotypic and molecular characterization of indigenous rhizobia nodulating chickpea in India. http://nopr.niscpr.res.in/handle/123456789/14128
Raverkar, K., Gupta, S. and Rao, D. 2005. Survival of soybean rhizobia in summer and proliferation during monsoon in vertisols of central India. Journal of the Indian Society of Soil Science, 53 (4), 591-597.
Rebecca, A.I.N., Hemamalini, V., Kumar, D., Srimathi, S., Muthumary, J. and Kalaichelvan, P. 2012. Endophytic Chaetomium sp. from Michelia champaca L. and its taxol production. J. Acad. Ind. Res, 1 (68), e72.
Rickert AA, Soria MA, Correa OS .2000. The adaptive acid response in Mesorhizobium sp World Journal of Microbiology and Biotechnology 16:475-480
Rodrigues, C.S., Laranjo, M. and Oliveira, S. 2006. Effect of heat and pH stress in the growth of chickpea mesorhizobia. Current Microbiology, 53, 1-7. https://doi.org/10.1007/s00284-005-4515-8
Sabri YAAA, Dizayee ATR .2023. The DRIS methodology to determine the best balance of Nitrogen, phosphorus and potassium levels on the yield of Chickpea plant (Cicer arietinum L.) Tikrit Journal for Agricultural Sciences 23:208-221. https://doi.org/10.25130/tjas.23.4.16
Sharma, Y.C., Singh, B. and Korstad, J. 2010. High yield and conversion of biodiesel from a nonedible feedstock (Pongamia pinnata). Journal of agricultural and food chemistry, 58 (1), 242-247. https://doi.org/10.1021/jf903227e
Singh, M., Kumar, K., Bisht, I.S., Dutta, M., Rana, M.K., Rana, J.C. 2015. Exploitation of wild annual Cicer species for widening the gene pool of chickpea cultivars. Plant Breeding, 134 (2), 186-192. https://doi.org/10.1111/pbr.12254.
Singh, N., Jain, P., Ujinwal, M. and Langyan, S. 2022. Escalate protein plates from legumes for sustainable human nutrition. Frontiers in nutrition, 9, 977986.
https://doi: 10.3389/fnut.2022.977986
Somasegaran, P. and Hoben, H.J. 2012. Handbook for rhizobia: methods in legume-Rhizobium technology. Springer Science & Business Media. https://doi: 10.1007/978-4613-8375-8
Strobel, G. 2018. The emergence of endophytic microbes and their biological promise. Journal of Fungi, 4 (2), 57. https://doi.org/10.3390/jof4020057.
Swarnalakshmi, K., Yadav, V., Tyagi, D., Dhar, D.W., Kannepalli, A. and Kumar, S. 2020. Significance of plant growth promoting rhizobacteria in grain legumes: Growth promotion and crop production. Plants, 9 (11), 1596. https://doi.org/10.3390/plants9111596.
Tarolli, P., Luo, J., Park, E., Barcaccia, G. and Masin, R. 2024. Soil salinization in agriculture: Mitigation and adaptation strategies combining nature-based solutions and bioengineering. Iscience, 27 (2). https://doi.org/10.3389/fmicb.2019.01885
Teja, M., Kumar, D.M., Bindiya, P. and Sudhakar, G. 2022. Microbial profiling, bioremediation of cadmium and dye decolourization competence of marine bacterial isolates. In New Innovations in Chemistry and Biochemistry, Book Publisher International (a part of SCIENCEDOMAIN International).
https://dio: 10.9734/bpi/nicb/v7/1811B
Vandana, U.K., Rajkumari, J., Singha, L.P., Satish, L., Alavilli, H., Sudheer, P.D. 2021. The endophytic microbiome as a hotspot of synergistic interactions, with prospects of plant growth promotion. Biology, 10 (2), 101. https://doi.org/10.3390/biology10020101.
Varshney, R.K., Roorkiwal, M., Sun, S., Bajaj, P., Chitikineni, A., Thudi, M. 2021. A chickpea genetic variation map based on the sequencing of 3,366 genomes. Nature, 599 (7886), 622-627. https://doi.org/10.6084/m9.figshare.16592819.
Vincent, J. and Humphrey, B. 1970. Taxonomically significant group antigens in Rhizobium. Microbiology, 63 (3), 379-382. https://doi.org/10.1099/00221287-63-3-379
Yadav, A.N. 2020. Plant microbiomes for sustainable agriculture: current research and future challenges. Springer. https://doi.org/10.1007/978-3-030-38453-1_16
Zhang, J., Wang, J., Zhu, C., Singh, R.P. and Chen, W. 2024. Chickpea: Its origin, distribution, nutrition, benefits, breeding, and symbiotic relationship with Mesorhizobium species. Plants, 13 (3), 429. https://doi.org/10.3390/plants13030429
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