Phenotyping Groundnut Rhizobia Native to Phosphorus Deficient Soils in Western Kenya

Velma Okaron^1, , Beatrice A. Were¹ and Benson O. Nyongesa¹

¹Department of Biological Sciences, University of Eldoret, P.O. Box 1125-30100, Eldoret, Kenya

Cite this paper:
Velma Okaron, Beatrice A. Were and Benson O. Nyongesa. Phenotyping Groundnut Rhizobia Native to Phosphorus Deficient Soils in Western Kenya. American Journal of Microbiological Research. 2017; 5(1):15-24. doi: 10.12691/ajmr-5-1-3

Abstract

Most soils in western Kenya are characterized by high acidity level and phosphorus (P) deficiency, which affect nodulation and nitrogen fixation of groundnut (Arachis hypogaea L.). This study aimed at characterising rhizobia capable of nodulating groundnut in P deficient soils in Western Kenya. Sixty four isolates out of the 68 were confirmed to be rhizobia due to their ability to nodulate groundnut. Ninety six percent of the isolates exhibited semi-globose to globose colony shape on yeast extract mannitol agar (YEMA). Groundnut was nodulated by both fast and slow growing rhizobia isolates with 81% being fast growers. Fifty one isolates representing 75% produced acid on YEMA medium supplemented with bromothymol blue (BTB). The isolates varied in their response to pH with 39 and 61 growing at pH 4.0 and 5.5, respectively. All the isolates grew at pH 7.0 and 8.5. YEMA medium containing glucose, sucrose, starch and citrate had 64, 61, 56 and 5 isolates growing, respectively. Sixty four isolates exhibited clear zone of solubilization on medium containing dicalcium phosphate as source of inorganic phosphate. Solubilization index (SI) varied from 1.1 to 6.8. Fast-growing rhizobia isolates N01, B02, I06, Q01, F05, C02, E01, Q03, I01 and B01 recorded the highest solubilization index of 3.8, 4.5, 4.6, 4.6, 4.7, 5.0, 5.1, 6.1, 6.1 and 6.8, respectively. Groundnut rhizobia showed variation in their potential to solubilize inorganic phosphate and effectively nodulate the host. The most promising isolates from this study would be used as bio-fertilizer upon further validation in the greenhouse and field.

Keywords:
legume nodulating bacteria characterization phosphorus deficiency arachis hypogaea inoculants

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Mokgehle, S. N., Dakora, F. D., & Mathews, C. Variation in N2 fixation and N contribution by 25 groundnut (Arachis hypogaea L.) varieties grown in different agro-ecologies, measured using 15 N natural abundance. Agriculture, Ecosystems & Environment, 195, 161-172. 2014.
[2]	Taurian, T., Ibañez, F., Fabra, A., & Aguilar, O. M. Genetic diversity of rhizobia nodulating Arachis hypogaea L. in central Argentinean soils. Plant and soil, 282(1-2), 41-52.2006.
[3]	Nekesa, P., Maritim, H. K., Okalebo, J. R., & Woomer, P. L. Economic analysis of maize-bean production using a soil fertility replenishment product (PREP-PAC) in western Kenya. African Crop Science Journal, 1999; 7(4): 585-590.
[4]	Owino, C. O., Owuor, P. O., & Sigunga, D. O. Elucidating the causes of low phosphorus levels in ferralsols of Siaya County, Western Kenya. Journal of Soil Science and Environmental Management, 2015; 6(9): 260-267.
[5]	Kisinyo, P. O., Othieno, C. O., Gudu, S. O., Okalebo, J. R., Opala, P. A., Ng'etich, W. K., & Opile, W. R. Immediate and residual effects of lime and phosphorus fertilizer on soil acidity and maize production in western Kenya. Experimental Agriculture, 2014; 50(1): 128-143.
[6]	Khan, M. S., Zaidi, A., Ahemad, M., Oves, M., & Wani, P. A. Plant growth promotion by phosphate solubilizing fungi–current perspective. Archives of Agronomy and Soil Science, 2010.; 56(1): 73-98.
[7]	Zhang, D., Zhang, C., Tang, X., Li, H., Zhang, F., Rengel, Z., & Shen, J. Increased soil phosphorus availability induced by faba bean root exudation stimulates root growth and phosphorus uptake in neighbouring maize. New Phytologist, 2016; 209(2): 823-831.
[8]	Marra, L. M., Oliveira, S. M., Soares, C. R. F. S., & Moreira, F. M. S. Solubilisation of inorganic phosphates by inoculant strains from tropical legumes. Science Agriculture (Piracicaba, Braz.), 2011; 68(5):.603-609.
[9]	Divito, G. A., & Sadras, V. O. How do phosphorus, potassium and sulphur affect plant growth and biological nitrogen fixation in crop and pasture legumes. A meta-analysis. Field Crops Research, 2014; 156:161-171.
[10]	Mathu, S., Herrmann, L., Pypers, P., Matiru, V., Mwirichia, R., & Lesueur, D. Potential of indigenous bradyrhizobia versus commercial inoculants to improve cowpea (Vigna unguiculata L. walp.) and green gram (Vigna radiata L. wilczek.) yields in Kenya. Soil science and plant nutrition, 2012; 58 (6):750-763.
[11]	Benson, O., Beatrice, A., Regina, N., Koech, P. K., Skilton, R. A., & Francesca, S. Morphological, genetic and symbiotic characterization of root nodule bacteria isolated from Bambara groundnuts (Vigna subterranea L. Verdc) from soils of Lake Victoria basin, western Kenya. Journal of Applied Biology and Biotechnology; 2015; 3(01): 1-10.
[12]	Jaetzold, R., Schmidt, H., Hornetz, B., & Shisanya, C.. Ministry of Agriculture Farm Management Handbook of Kenya VOL. II-Part C Subpart C1. Nairobi, Kenya: Ministry of Agriculture, 2006.
[13]	Muhati, S. I., Shepherd, K. D., Gachene, C. K., Mburu, M. W., Jones, R., Kironchi, G. O., & Sila, A. Diagnosis of soil nutrient constraints in small-scale groundnut (Arachis hypogaea L.) production systems of Western Kenya using infrared spectroscopy. Journal of Agricultural Science and Technology 2011; 111-127.
[14]	Okalebo, J. R., Gathua, K. W., & Woomer, P. L. Laboratory methods of plant and soil analysis:a working manual. Tropical Soil Biology and Fertility Programme, Nairobi. 2002
[15]	Hue, Andrade DS, Chueira LM. Isolation and characterization of new efficient and competitive bean (Phaseolus vulgaris L.) rhizobia in Brazil. Soil Biology and Biochemistry, 2000; 32: 1515-1528.
[16]	Vincent, J. M. A manual for the practical study of the root-nodule bacteria. (IBP Handbuch No. 15 des International Biology Program, London . XI u. 164 S., 10 Abb., 17 Tab., 7 Taf. Oxford-Edinburgh: Blackwell Scientific Publ., 45 s.1970.
[17]	Gupta, A., Gupta, A. K., Mahajan, R., Singh, D., Khosla, K., Lal, R., & Gupta, V. Protocol for isolation and identification of Agrobacterium Isolates from Stone Fruit plants and sensitivity of native A. tumefaciens isolates against agrocin produced by A. radiobacter strain K84. National Academy Science Letters, 2013; 36(1): 79-84.
[18]	Huang, B,., Lü, C., Wu, B. & Fan, L. A rhizobia strain isolated from root nodule of gymnosperm Podocarpus macrophyllus. Science China Serie. C: Life Science,2007; 50:1-6.
[19]	Mehta, S., & Nautiyal, C. S. An efficient method for qualitative screening of phosphate-solubilizing bacteria. Current microbiology, 2001; 43(1): 51-56.
[20]	Alikhani, H. A., Saleh-Rastin, N., & Antoun, H. Phosphate solubilization activity of rhizobia native to Iranian soils. In First international Meeting on microbial phosphate solubilization , 35-41. Springer Netherlands. 2006.
[21]	Laurette, N. N., Maxémilienne, N. B., Henri, F., Souleymanou, A., Kamdem, K., Albert, N. & François-Xavier, E. Isolation and Screening of Indigenous Bambara Groundnut (Vigna Subterranea) Nodulating Bacteria for their Tolerance to Some Environmental Stresses. American Journal of Microbiological Research, 2015; 3(2): 65-75.
[22]	Somasegaran, P., & Hoben, H. J. (1994). Collecting nodules and isolating rhizobia. In Handbook for Rhizobia (pp. 7-23). Springer New York
[23]	Team, R. C. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. 2015. URL h ttp. www. R-project. org. Accessed September .13, 2016
[24]	Muthini, M., Maingi, J. M., Muoma, J. O., Amoding, A., Mukaminega, D., Osoro, N. & Ombori, O.Morphological assessment and effectiveness of indigenous rhizobia isolates that nodulate P. vulgaris in water hyacinth compost testing field in Lake.
[25]	Sharma, M. P., Srivastava, K., & Sharma, S. K. Biochemical characterization and metabolic diversity of soybean rhizobia isolated from Malwa region of central India. Plant Soil Environ, 56(8), 375-383.2010Victoria Basin. British Journal of Applied Science & Technology,2014; 4(5): 718.
[26]	Sanginga, N., Danso, S. K. A., & Bowen, G. D. Nodulation and growth response of Allocasuarina and Casuarina species to phosphorus fertilization. Plant and Soil, 1989; 118 (1-2): 125-132.
[27]	Teixeira, F.C.P., Borges, W. L., Xavier, G. R, Rumjanek, N. G. Characterization of indigenous rhizobia from Caatinga. Brazilian Journal of Microbiology, 2010; 41: 201-208.
[28]	Boakye, E. Y., Lawson, I. Y. D., & Danso, S. K. A. Characterization and diversity of rhizobia nodulating selected tree legumes in Ghana. Symbiosis, 69 (2), 89-99. 2016
[29]	Kapembwa, R., Mweetwa, A. M., Ngulube, M., & Yengwe, J. Morphological and Biochemical Characterization of Soybean Nodulating Rhizobia Indigenous to Zambia. Journal of Agricultural Research, 2016; 5: 84.
[30]	Sprent, J.I. Evolution and diversity in the legume-rhizobium symbiosis: chaos theory? Plant Soil, 1994; 161:1-10.
[31]	Sayyed, R. Z., Jamadar, D. D., & Patel, P. R. Production of Exo-polysaccharide by Rhizobium sp. Indian journal of microbiology, 2011; 51(3): 294-300.
[32]	Batista, J. S. S., Hungria, M., Barcellos, F. G., Ferreira, M. C., & Mendes, I. C. Variability in Bradyrhizobium japonicum and B. elkanii seven years after introduction of both the exotic microsymbiont and the soybean host in a Cerrados soil. Microbial ecology, 2007; 53(2): 270-284.
[33]	Wahab, A. A., Zahran, H. H., & Abd-Alla, M. H. Root-hair infection and nodulation of four grain legumes as affected by the form and the application time of nitrogen fertilizer. Folia microbiologica, 199 41(4), 303-308. 1996.
[34]	Sobczak, I., & Lolkema, J. S. The 2-hydroxycarboxylate transporter family: physiology, structure, and mechanism. Microbiology and Molecular Biology Review, 2005; 69: 665-695.
[35]	Küçük, Ç., Kivanç, M., & Kinaci, E. Characterization of Rhizobium sp. isolated from bean. Turkish Journal of Biology, 2000; 30(3): 127-132.
[36]	Hungria, M., de S Andrade, D., de O Chueire, L. M., Probanza, A., Guttierrez-Mañero, F. J., & Megı́as, M. Isolation and characterization of new efficient and competitive bean (Phaseolus vulgaris L.) rhizobia from Brazil. Soil Biology and Biochemistry, 2000; 32, 1515-1528.
[37]	Brígido, C., & Oliveira, S. Most acid-tolerant chickpea mesorhizobia show induction of major chaperone genes upon acid shock. Microbial Ecology, 2013; 65: 145-153.
[38]	Rodrigues, C. S., Laranjo, M., & Oliveira, S. Effect of heat and pH stress in the growth of chickpea mesorhizobia. Current Microbiology, 2006; 53(1): 1-7.
[39]	Kumari, B. S., Ram, M. R., & Mallaiah, K. V. Studies on exopolysaccharide and indole acetic acid production by Rhizobium strains from Indigofera. African Journal of Microbiology Research, 2009; 3(1): 10-14.
[40]	Kumar, G. K., & Ram, M. R. Phosphate solubilizing rhizobia isolated from Vigna trilobata. American Journal of Microbiological Research, 2014; 2(3): 105-109.
[41]	Sujatha, E., Grisham, S., Reddy, S. M. Phosphate solubilization by thermophillic microorganisms. Indian Journal of Microbiology, 2004; 44: 101–104.
[42]	Harun, M. Sattar, M. A., Uddin, M. I. and Young, J. P. W. Molecular characterization of symbiotic root nodulating rhizobia isolated ntifrom lentil (Lens culinaris Medik.). Journal of Environmental Agriculture and Food Chemistry, 2009; 8: 602-612.