The influence of rhizobia and Arbuscular mycorrhizal fungi co-inoculation on growth and yield of Chickpea (Cicer arietinum) in the greenhouse and the field

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Research Paper 30/07/2022
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The influence of rhizobia and Arbuscular mycorrhizal fungi co-inoculation on growth and yield of Chickpea (Cicer arietinum) in the greenhouse and the field

Hizkiyas Wondosen, Beyene Dobo, Abraham Mikru, Kedir Woliy
Int. J. Biosci.21( 1), 268-286, July 2022.
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The use of chemical inputs in agriculture has a negative influence on the environment. Application of chemical fertilizers (F) can be reduced by using bio-fertilizers like arbuscular mycorrhizal fungus (AMF) and Rhizobium (R). Chick pea was co-inoculated and grown in the greenhouse and in the field to investigate their effect on growth and yield. All assessed plant growth parameters were found to be highest in plants treated with AMF and Rhizobium after three months of growth. Plants treated with R+AMF+F had greater average shoot length, stem girth, shoot dry weight, root dry weight, shoot fresh weight, branch per plant, and number of pods than those treated with R+M. However, R+M had a higher number of nodules than R+AMF+F in the greenhouse, and for co-inoculation in the field, indigenous rhizobium and AMF with fertilizer application had the highest shoot length (82.0±2.3), number of pods (75.0±2.6), dry weight (109.5±11.2), 100 seed weight (14.2±0.2), and husk weight (12.06±0.2) when compared to the control. Furthermore, root colonization ranged from 63.7±2.9% to 85.0±8.1% for all AMF infected treatments. In the green house, the maximum spore density (560.0±17.3 spore/50g dry soil) was recorded for solitary mycorrhiza inoculated chickpea. As a result, it is determined that dual inoculation of the pulse with AMF and Rhizobium, both with and without NPK treatment, increased growth and production. This suggests that smallholder farmers may be able to use bio-inoculants to boost chickpea output.


Abdiani SA, Kakar K, Gulab G, Aryan S. 2019. Influence of bio fertilizer application methods on growth and yield performances of green pepper. SSRN Electron Journal 2, 68 – 74.

Alireza Tavasolee, Nasser Aliasgharzad, Gholamreza SalehiJouzani, Mohsen Mardi, Ahmad Asgharzadeh. 2011. Interactive effects of Arbuscular mycorrhizal fungi and rhizobial strains on chickpea growth and nutrient content in the plant. African Journal of Biotechnology 10, 7585-7591.

Arumugam R, Rajasekaran S, Nagarajan SM. 2010. Response of Arbuscular Mycorrhizal fungi and Rhizobium inoculation on growth and chlorophyll content of Vigna unguiculata (L) Walp Var. Pusa 151. Journal of Applied Science and Environmental Management 14, 113 – 115.

Azcon-Aguilar C, Jaizme-Vega MC, Calvet C. 2002. The Contribution of Arbuscular Mycorrhizal Fungi to the Control of Soil-borne Plant Pathogens. In: Gianinazzi S, Schuepp H, editors. Mycorrhizal Technology: from Genes to Bioproducts-Achievements and Hurdles in Arbuscular Mycorrhiza Research. Basel 187-198.

Bala A, Abaidoo R, Woomer P. 2010. Rhizobia Strain Isolation and Characterization Protocol. URL: www. N2Africa. org (accessed on May 2019).

Barea JM. 2000. Rhizosphere and Mycorrhizae of Field Crops. In: Toutant P, Balazs E, Galante E, et al., editors. Biological Resource Management, Connecting Science and Policy (OECD) (Berlin, Heidelberg, New York: INRA and Springer. DOI: 10.1631/jzus.2006.B0503

Beaty RD, Kerber JD. 1993. Concepts, Instrumentation, and Techniques in Atomic Absorption Spectrophotometry. 2nd Edition. The Perkin-Elmer Corporation, Norwalk, CT, USA. DOI: 10.1080/03650340.2016.114764

Beyene Dobo, Fasil Asefa, Zerihun Asfaw. 2016. Response of Selected Perennial Crops and Trees to AMF Inoculation and NPK Application in Pot Culture. Academic Journal of Biotechnology 4, 325-336. DOI: 10.15413/ajb.2016.0119

Bhattacharjee S, Sharma GD. 2012. Effect of dual inoculation of Arbuscular Mycorrhiza and Rhizobium on the Chlorophyll, Nitrogen, and Phosphorus Contents of Pigeon Pea. Advances in Microbiology 2, 561-564. DOI: 10.4236/aim.2012.24072

Black C, Evans AD, White JL, Ensminger LE, Clark FE. 1965. Methods of Soil Analysis Part 1. Physical and Mineralogical Properties Including Statistics of Measurement and Sampling, American Society of Agronomy, Madison, Wis, USA.

Botir Khaitov, Akhmad Kurbonov, Anvar Abdiev, Maksud Adilov. 2016. Effect of chickpea in association with Rhizobium to crop productivity and soil fertility. Eurasian Journal of Soil Science 5(2), 105-112. DOI: 10.18393/ejss.2016.2.105-112

Bournaud C, James EK, Faria SM, Lebrun M, Melkonian R, Duponnois R, Tisseyre P, Moulin L, Prin Y. 2018. Interdependency of efficient nodulation and arbuscular mycorrhization in Piptadenia gonoacantha, a Brazilian legume tree. Plant, Cell & Environment 41, 2008 – 2020.

Cavell AJ. 1995. The colorimetric determination of phosphorus in plant materials. Journal of the Science of Food and Agri­culture 6, 479-480.

Chalk PMR, Souza S, Urquiaga BJ, Alves R, Boddey RM. 2006. The role of Arbuscular mycorrhizae in legume symbiotic performance. Soil Biology and Biochemistry 38, 2944-2951.

Egamberdieva D, Shurigin V, Gopalakrishnan S, Sharma R. 2014. Growth and symbiotic performance of chickpea (Cicer arietinum) cultivars under saline soil conditions. Journal of Biological and Chemical Research 31(1), 333-341.

Gerdemann JW, Nicolson TH. 1963. Spores of mycorrhizal Endogone species extracted from soil by wet sieving and decanting. Transactions of the British Mycological Society 46, 235-244.

Hassen AI, Bopape FL, Trytsman M. 2014. Nodulation Study and Characterization of Rhizobial Microsymbionts of Forage and Pasture Legumes in South Africa. World 2, 93-100. DOI: 10.12691/wjar-2-3-2

Jordan DC. 1982. Transfer of Rhizobium japonicum Buchanan 1980 to Bradyrhizobium genus of slow-growing, root nodule bacteria from leguminous plants. International Journal of Systematic Bacteriology 32, 136-139.

Kaschuk G, Kuyper TW, Leffelaar PA, Hungria M, Giller KE. 2009. Are the rates of photosynthesis stimulated by the carbon sink strength of rhizobial and arbuscular mycorrhizal symbioses?. Soil biology & biochemistry 41, 1233 – 1244.

Kathiresan K, Selvam MM. 2006. Evaluation of beneficial bacteria from mangrove soil. Botanica Marina 49, 86-88. DOI: 10.1515/BOT.2006.011

Khan IA, Ayub N, Mirza SN, Nizam SM, Azam M. 2008. Synergistic effect of dual inoculation (Vesicular-Arbuscular Mycorrhizae) on the growth and nutrient uptake of Medicago sativa. Pakistan Journal of Botany 40, 939-945.

Kossmann and Valentine. 2014. The reallocation of carbon in P deficient lupins affects biological nitrogen fixation. Journal of Plant Physiology 171, 1619-1624. DOI: 10.1016/j.jplph.2014.07.017

Larimer AL, Clay K, Bever JD. 2014. Synergism and context dependency of interactions between arbuscular mycorrhizal fungi and rhizobia with a prairie legume. Ecology 95, 1045-1054. DOI: 10. 1890/13-0025.1

Lupwayi N, Haque I. 1994. Legume-Rhizobium Technology Manual. Environmental Sciences Division International Livestock Center for Africa, Addis Ababa, Ethiopia, 1-93.

Malina SF, Sharma GD. 2013. Effects of Mycorrhizal Fungi and Rhizobia Inoculation on the Growth of Crotalaria Pallida- An Approach to Tripartite Symbiosis. Life Science 2(6), 112 – 114.

McGonigle TP, Miller HM, Evans GD, Fairchild LG, Swan AJ. 1990. A new method that gives an objective measure of colonization of roots by vesicular-arbuscular mycorrhizal fungi. New Phytologist 115, 495-501. DOI: 10.1111/j.1469-8137. 1990.tb00476.x

Mensah JA, Koch AM, Antunes PM, Kiers ET, Hart M, Bücking H. 2015. High functional diversity within species of arbuscular mycorrhizal fungi is associated with differences in phosphate and nitrogen uptake and fungal phosphate metabolism. Mycorrhiza 2, 1-14. DOI: 10.1007/s00572-015-0631-x

Mhango J, Mahamadi D, Ndusa B, Woomer PL, Abaidoo R, Karanja NK, Uwizerwa M. 2013. Rhizobium collection, testing, and the identification of candidate elite strains.

Mortimer PE, Pe´rez-Ferna´ ndez MA, Valentine AJ. 2008. The role of arbuscular mycorrhizal Colonization in the carbon and nutrient economy of the tripartite symbiosis with Nodulated Phaseolus vulgaris. Soil Biology and Biochemistry 40, 1019-1027. DOI: 10.1016/j.soilbio.2007.11.014

Murat E, Demir S, Tufenkci EOS, Oguz F, Akkopru A. 2011. Effects of Rhizobium, Arbuscular mycorrhizae, and whey applications on some properties in chickpea (Cicer Arietinum L.) under irrigated and rainfed conditions. Field Crops Research 122, 14-24. DOI: 10.1016/j.fcr.2011.02.002

Muthini Morris, Maingi John, Muoma John, Amoding Alice. 2014. Morphological Assessment and Effectiveness of Indigenous Rhizobia Isolates that Nodulate P. vulgaris in water Hyacinth Compost Testing Field in Lake Victoria Basin. Journal of Applied Science and Technology 4, 718-738. DOI: 10.9734/BJAST/2014/5757

Nabrzyski M, Gajewska R. 1998. Aluminium determination in foods by using Spectrophotometric oxine and flame AAS methods. Molecular Nutrition and Food Research 42, 109-111. DOI: 10.1002/(SICI)1521-3803(199804)42:02

Nandwani R, Dudeja SS. 2009. Molecular diversity of a native Mesorhizobial the population of nodulating chickpea (Cicer arietinum L.) in Indian soils. Journal of Basic Microbiology 49, 463-470. DOI: 10.1002/jobm. 200800355

Plenchette CA, Fortin A, Forlan N. 1983. Growth response of several plant species to Mycorrhiza in a soil of moderate P-fertility. I. Mycorrhizae under field conditions. Plant Soil 70, 199-203.

Püschel D, Janoušková M, Hujslová M, Slavíková R, Gryndlerová H, Jansa J. 2017. Plant-fungus competition for nitrogen erases mycorrhizal growth benefits of Andropogon gerardii under limited nitrogen supply. Ecology and Evolution 6, 4332-4346. DOI: 10.1002/ece3.2207

Rodriguez-Navarro D, Buendia A, Camacho M, Lucas M, Santamaria C. 2000. Characterization of Rhizobium spp. bean isolates from South-West Spain. Soil Biology and Biochemistry 32, 1601-1613. DOI: 10.1016/S0038-0717(00)00074-2

Sharma K, Kumawat D. 2011. A study on evaluation of nitrogen fixation potential in soybean cultivar using commercial and indigenous strains. European Journal of Experimental Biology 1, 93-97.

Shurigin V, Davranov K, Abdiev A. 2015. Screening of salt-tolerant rhizobia for improving growth and nodulation of chickpea (Cicer arietinum) under arid soil conditions of Uzbekistan. Journal of Biological and Chemical Research 32(2), 534-540.

Silveira PD, Cardoso EJ. 2004. Arbuscular Mycorrhizal and kinetic parameters of Phosphorus absorption by bean plants. Agricultural Science 61, 203-209. DOI: 10.1590/S0103-90162004000200013

Simon Z, Mtei K, Gessesse A, Ndakidemi PA. 2014. Isolation and Characterization of Nitrogen-Fixing Rhizobia from Cultivated and Uncultivated Soils of Northern Tanzania. American Journal of Plant Science 5, 4050 – 4056. DOI: 10.4236/ajps.2 014.526423

Singh PK, Singh AK. 2010. Response of mungbean (Vigna radiata L. Wilczek) concerning micronutrients content and yield as affected by sulfur and cobalt application. Environment and Ecology 28, 564-568.

Smith SE, Read DJ. 1979. Mycorrhizal Symbiosis. Academic, San Diego, CA.

Solomon Legesse, Fasil Asefa. 2014. Symbiotic and Phenotypic Characteristics Of Rhizobia Nodulating Faba Bean (Vicia Faba) From Tahtay Koraro, Northwestern Zone Of Tigray Regional State, Ethiopia. International journal of technology enhancements and emerging engineering research 2, 15-23.

Somasegaran P, Hoben HJ. 1994. Handbook for Rhizobia: Methods in Legume Rhizobium Technology. Springer-Verlag New York, USA, 450.

Somasegaran P, Hoben HJ. 2012. Handbook for Rhizobia: Methods in Legume Rhizobium Technology. Springer Science and Business Media, Berlin.

Spaink HP, Kondorosi A, Hooykaas PJJ. 1998. The Rhizobiaceae (Kluwer Academic Publishers, Dordrecht, The Netherlands.

Sylvia DM, Jarstfer AG. 1994. Production of Inoculum and Inoculation with Arbuscular Mycorrhizal Fungi. In: Robson AD, Abbott LK, Malajczuk N, editors. Management of Mycorrhizas in Agriculture, Horticulture, and Forestry 231-238.

Thenua OVS, Singh SP, Shivakumar BG. 2010. Productivity and economics of chickpea (Cicer arietinum) fodder sorghum (Sorghum bicolor) cropping system as influenced by P sources, bio-fertilizers, and irrigation to chickpea. Indian Journal of Crop Science 12, 253-261.

Utobo EB, Ogbodo EN, Nwogbaga AC. 2011. Techniques for Extraction and Quantification of Arbuscular Mycorrhizal Fungi. Libyan Agriculture Research Center Journal International 2, 68-78.

Vincent JM. 1970. A manual for the practical study of root-nodule bacteria, Blackwell Scientific, Oxford. I.B.P handbook.

Yinsuo J, Vincent MG, Colin JS. 2004. The Influence of Rhizobium and Arbuscular Mycorrhizal Fungi on Nitrogen and Phosphorus Accumulation by Vicia faba. Annals of Botany 94, 251-258. DOI: 10.1093/aob/mch135

Zelalem Kassa. 2018. Response of Lentil (Lens culinaris Medik) Varieties to Rhizobium Inoculation and Inorganic Fertilizer Application in Siya Debrina Wayu District, North Shewa Zone, Ethiopia. MSc Thesis. Debre Berhan, Debre Berhan, Ethiopia.