Skip to content
BY 4.0 license Open Access Published by De Gruyter Open Access July 19, 2019

Isolation, characterization and purification of Rhizobium strain to enrich the productivity of groundnut (Arachis hypogaea L.)

  • Akbar Hossain EMAIL logo , Sunil Kumar Gunri , Manashi Barman , Ayman EL Sabagh and Jaime A. Teixeira da Silva
From the journal Open Agriculture


Groundnut (Arachis hypogaea L.) is an important food legume in tropical and subtropical areas because of its ability to adapt to a wide range of agro-climatic regions. Groundnut is usually cultivated in nutrient-poor soil and rain-fed conditions, so average yield tends to be very low relative to potential yield. Even though the nitrogen (N) requirement of groundnut is much higher than cereals due to its high protein content, it has the capacity to meet 60-80% of N-based requirements through symbiotic N fixation via its root nodules. In its symbiotic relationship with legumes, Rhizobium fixes N, thereby positively impacting the content of this nutrient. This study aimed to isolate, characterize and purify microbial strains of Rhizobium specific to groundnut in a bid to increase this legume’s productivity. The research was conducted in the AICRP-Groundnut laboratory and greenhouse of the Directorate of Research, BCKV, in Kalyani, India during October 2016 to March 2017. Two Rhizobium isolates (RhBC and NRA1) were isolated and selected from groundnut pot cultures. After 45 days, NRA1 produced higher plant biomass, longer roots and shoots, more nodules and higher nodule dry weight than RhBC. NRA1 was selected for a future field trial. The two isolated microbial strains will aid in the screening of additional local isolates to test their effectiveness when co-cultured with local groundnut cultivars to increase yield in soil with low fertility.


[1] Andrews M., Andrews M.E., Specificity in legume-rhizobia symbioses. Int. J. Mol. Sci., 2017, 18(4), 705. doi:10.3390/ijms1804070510.3390/ijms18040705Search in Google Scholar

[2] Arora D.R., The Text Book of Microbiology. New Delhi: CBS Publisher, 2003, pp. 41-48Search in Google Scholar

[3] Badawi F.S.F., Biomy A.M.M., Desoky A.H., Peanut plant growth and yield as influenced by co-inoculation with Bradyrhizobium and some rhizo-microorganisms under sandy loam soil conditions. Ann. Agric. Sci., 2011, 56(1), 17–2510.1016/j.aoas.2011.05.005Search in Google Scholar

[4] Badigannavar A.M., Kale D.M., Murty G.S.S., Assessment of yielding ability of Trombay groundnut varieties through growth analysis. J. Oilseeds Res., 2002, 19, 38–43Search in Google Scholar

[5] Bailey W.K., Hammons R.O., Registration of Chico Peanut Germplasm1 (Reg. No. GP 2). Crop Sci., 1975, 15(1), 105–10510.2135/cropsci1975.0011183X001500010050xSearch in Google Scholar

[6] BCKV (Bidhan Chandra Krishi Viswavidyalaya)., AICRP-Groundnut, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya Kalyani, Nadia, West Bengal, India., 2016Search in Google Scholar

[7] Bhuiyan M.A.H., Rahman M.M., Khanam D., Khaleque M.A., Nodulation and yield of groundnut in relation to strains of Bradyrhizobium. Bangladesh J. Sci. Indust. Res., 1998, 33(3), 323–326Search in Google Scholar

[8] Biswas P., Hosain D., Ullah M., Akter N., Bhuiya M.A.A., Performance of groundnut (Arachis hypogaea L.) under different levels of bradyrhizobial inoculum and nitrogen fertilizer. SAARC J. Agric., 2003, 1, 61–68Search in Google Scholar

[9] Brahmaprakash G.P., Girisha H.C., Navi V., Laxmipathy R., Hegde SV., Liquid Rhizobium inoculant formulations to enhance biological nitrogen fixation in food legumes. J. Food Legumes, 2007, 20(1), 75–79Search in Google Scholar

[10] de Rosalia C.E., Santos S., da Silva V.S., de Freitas A.D., da Silva A.F., de Vasconcelos Bezerra R., da Silva Ferreira J., Prospecting of efficient rhizobia for peanut inoculation in a Planosol under different vegetation covers. African J. Microb. Res., 2017, 11(4), 123–131Search in Google Scholar

[11] Deshwal V.K., Dubey R.C., Maheshwari D.K., Isolation of plant growth-promoting strains of Bradyrhizobium (Arachis) sp. with biocontrol potential against Macrophomina phaseolina causing charcoal rot of peanut. Curr Sci., 2003, 84(3), 443–448Search in Google Scholar

[12] Dinesh R., Anandaraj M., Kumar A., Bini Y.K., Subila K.P., Aravind R., Isolation, characterization, and evaluation of multi-trait plant growth promoting rhizobacteria for their growth promoting and disease suppressing effects on ginger. Microbiol. Res., 2015, 173, 34–4310.1016/j.micres.2015.01.014Search in Google Scholar

[13] El-Ghandour I.A., Galal Y.G.M., Soliman S.M., Yield and N2 fixation of groundnut (Arachis hypogaea L.) in response to inoculation with selected Bradyrhizobium strains and mycorrhizal fungi. Egyptian J. Microbiol., 1997, 32(4), 467–480Search in Google Scholar

[14] Faddin M.J.F., Biochemical tests for identification of medical bacteria, 3rd ed. Lippincott Williams&Wilkins, Philadelphia, PA, 2000, pp. 363–367Search in Google Scholar

[15] Erum S., Asghari B., Variation in phytohormone production in Rhizobium strains at different altitudes of northern areas of Pakistan. Int. J. Agric. Biol., 2008, 10, 536–540Search in Google Scholar

[16] FAO (Food and Agricultural Organization)., Data., 2018. Accessed on June 25, 2019Search in Google Scholar

[17] Fentahun M., Akhtar M.S., Muleta D., Lemessa F., Isolation and characterization of nitrogen deficit Rhizobium isolates and their effect on growth of haricot bean. African J. Agric. Res., 2013, 8(46), 5942–5952Search in Google Scholar

[18] Gouda S., Kerry R.G., Das G., Paramithiotis S., Shin H.S., Patra J.K., Revitalization of plant growth promoting rhizobacteria for sustainable development in agriculture. Microbiol. Res., 2017, 206, 131–14010.1016/j.micres.2017.08.016Search in Google Scholar

[19] Gunri S.K., Nath R., Gaintait S., Barman M., Paul S., Puste A.M., Bera, P.S., Optimization of kharif groundnut (Arachis hypogaea L.). In: Proceedings of International Symposium on “Eco-Efficiency in Agriculture and Allied Research”, 20–23 January, 2017. Organized by CWSS-BCKV at FACC, Kalyani, Nadia, WB, India, 2017, pp. 281–282Search in Google Scholar

[20] Hasan M., Sahid I.B., Evaluation of Rhizobium inoculation in combination with phosphorus and nitrogen fertilization on groundnut growth and yield. J. Agron., 2016, 15(3), 142-14610.3923/ja.2016.142.146Search in Google Scholar

[21] Hemraj V., Diksha S., Avneet G., A review on commonly used biochemical test for bacteria. Innovare J. Life Sci., 2013, 1(1), 1-7Search in Google Scholar

[22] IRRI (International Rice Research Institute)., Biometrics and Breeding Informatics: Products STAR Statistical Tool for Agricultural Research. Los Baños, Laguna, Philippines, 2014. Accessed on 07 April 2019Search in Google Scholar

[23] Ishihara M., Matsunaga M., Hayashi N., Tišler V., Utilization of D-xylose as carbon source for production of bacterial cellulose. Enzyme Microbial. Technol., 2002, 31(7), 986–99110.1016/S0141-0229(02)00215-6Search in Google Scholar

[24] Kale D.M., Mouli C., Murty G.S.S., Rao M.V.P., Development of a new groundnut variety, TG-26, by using induced mutations in cross breeding. Mutation Breed. Newslett., 1997, 43, 25–27Search in Google Scholar

[25] Kale DM, Murty GSS, Badigannavar AM, Dhal JK (2009) New Trombay groundnut variety TG 51 for commercial cultivation in India. J. SAT Agric. Res., 2009, 7. Accessed on June 25, 2019Search in Google Scholar

[26] Kenasa G., Jida M., Assefa F., Characterization of phosphate solubilizing faba bean (Vicia faba L.) nodulating rhizobia isolated from acidic soils of Wollega, Ethiopia. Sci. Technol, Arts Res. J., 2014, 3(3), 11–1710.4314/star.v3i3.2Search in Google Scholar

[27] Kneen B.E., LaRue T.A., Congo red absorption by Rhizobium leguminosarum. Appl. Environ. Microbiol., 1983, 45(1), 340-34210.1128/AEM.45.1.340-342.1983Search in Google Scholar PubMed PubMed Central

[28] Koser S.A., Utilization of the salts of organic acids by the colonaerogenes group. J. Bacteriol., 1923, 8(5), 493–52010.1128/JB.8.5.493-520.1923Search in Google Scholar PubMed PubMed Central

[29] Kumar A., Meena R.N., Yadav L., Gilotia Y.K., Effect of organic and inorganic sources of nutrient on yield, yield attribute and nutrient uptake of rice cv. PRH-10. The Bioscan, 2014, 9(2), 595–597Search in Google Scholar

[30] Küpper F.C., Feiters M.C., Olofsson B., Kaiho T., Yanagida S., Zimmermann M.B., Carpenter L.J., Luther III G.W., Lu Z., Jonsson M., Kloo L., Commemorating two centuries of iodine research: an interdisciplinary overview of current research. Angew. Chemie Int. Ed., 2011, 50(49), 11598–1162010.1002/anie.201100028Search in Google Scholar PubMed

[31] Lamas F.M., Mercante F.M., Otsubo A.A., Passos D.P., Use of inoculant for groundnuts: effects on productivity. Comunicado Técnico - EMBRAPA Agropecuária Oeste, 2000, 28: 5 pp. (in Portuguese with English abstract).Search in Google Scholar

[32] Lowe G.H., The rapid detection of lactose fermentation in paracolon organism by demonstration of 6-D-galactosidase. J. Med. Lab. Technol., 1962, 19, 21–25Search in Google Scholar

[33] MacFaddin J.F., Biochemical tests for Identification of Medical bacteria. Williams and Wilkins Co., Baltimore, USA, 1976, 51–54Search in Google Scholar

[34] Manasa K., Reddy R.S., Triveni S., Kumar B.K., Priya N.G., Characterization of Rhizobium isolates and their potential PGPR characteristics of different rhizosphere soils of Telangana region, India. Int. J. Curr. Microbiol. Appl. Sci., 2017, 6(5), 2808–281310.20546/ijcmas.2017.605.316Search in Google Scholar

[35] McDevitt S., Methyl red and Voges Proskauer test protocols. American Society for Microbiology, 2009Search in Google Scholar

[36] Ondieki D.K., Nyaboga E.N., Wagacha, J.M., Mwaura F.B. Morphological and genetic diversity of Rhizobia nodulating cowpea (Vigna unguiculata L.) from agricultural soils of lower eastern Kenya. Int. J. Microbiol., 2017, Article ID 8684921, 9 pp. doi:10.1155/2017/868492110.1155/2017/8684921Search in Google Scholar PubMed PubMed Central

[37] Pandeeswari N., Kalaiarasu S., Performance of salt tolerant mutant Rhizobia inoculation with graded levels of nitrogen on the growth and yield attributes of groundnut (Arachis hypogaea L.). Int. J. Recent Sci. Res., 2017, 8(4), 16275–1627910.24327/ijrsr.2017.0804.0117Search in Google Scholar

[38] Pelczar Jr. M.J., Reid R.D., Chan E.C.S., Microbiology, 4th Edition. Tata McGraw-Hill Publishing Company Ltd, New Delhi, 1977Search in Google Scholar

[39] Pulatova D.Z., Saimnazarov U.B., Bakhromov I.U., Kozhemyakov A.P., Provorov N.A., Selection of nodule bacteria strains forming effective symbiosis with the peanut under central Asian conditions. Russian Agric. Sci., 1999, 3, 20–24Search in Google Scholar

[40] Rahman M.A., Effect of calcium and Bradyrhizobium inoculation of the growth, yield and quality of groundnut (Arachis hypogaea L.). Bangladesh J. Sci. Indust. Res., 2006, 41, 181–18810.3329/bjsir.v41i3.288Search in Google Scholar

[41] Russell A.D., Hugo W.B., Ayliffe G., Principles and Practice of Disinfection, Sterilization and Preservation. Blackwell Scientific Publications. Oxford, 1982Search in Google Scholar

[42] Sadowsky M.J., Keyser H.H., Bohlool B.B., Biochemical characterization of fast- and slow-growing rhizobia that nodulate soybeans. Int. J. Syst. Bacteriol., 33(4), 716–72210.1099/00207713-33-4-716Search in Google Scholar

[43] Sajid M., Rab A., Wahid F., Shah S.N.M., Jan I., Khan M.A., Hussain S.A., Khan M.A., Iqbal Z., Influence of rhizobium inoculation on growth and yield of groundnut cultivars. Sarhad J. Agric., 2011, 27(4), 573–576Search in Google Scholar

[44] Shahzad F., Shafee M., Abbas F., Babar S., Tariq M.M., Ahmad Z., Isolation and biochemical characterization of Rhizobium meliloti from root nodules of alfalfa (Medico sativa L.). J. Anim. Plant Sci., 2012, 22(2), 522–524Search in Google Scholar

[45] Sharma P., Sardana V., Kandola S.S., Response of groundnut (Arachis hypogaea L.) to Rhizobium inoculation. Libyan Agric. Res. Center J. Int., 2011, 2, 101–104Search in Google Scholar

[46] Singh A.L., Basu M.S., Singh N.B., Potential of groundnut in North Eastern States of India. National Research Centre for groundnut (ICAR), Junagadh, India, 2003, 75 pp.Search in Google Scholar

[47] Singh B., Kaur R., Singh K., Characterization of Rhizobium strain isolated from the roots of Trigonella foenumgraecum (fenugreek). African J. Biotechnol., 2008, 7(20), 3671–3676Search in Google Scholar

[48] Somasegaran P., Hoben J., Methods in legume-Rhizobium technology, University of Hawaii NifTAL Project and Mircen, Hawaii Institute of Tropical Agriculture and Human Resources, College of Tropical Agriculture, 1985, 367 pp.Search in Google Scholar

Received: 2018-11-16
Accepted: 2019-05-18
Published Online: 2019-07-19

© 2019 Akbar Hossain et al., published De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 Public License.

Downloaded on 30.11.2023 from
Scroll to top button