Exploration of indigenous free nitrogen-fixing bacteria from rhizosphere of Vigna radiata for agricultural land treatment


  • Novi Arfarita Universitas Islam Malang (UNISMA)
  • Anton Muhibuddin Faculty of Agriculture, University of Brawijaya
  • Tsuyoshi Imai Graduate School of Science and Engineering, Faculty of Engineering, Yamaguchi University




biofertilizer, indigenous bacteria, nitrogen-fixing bacteria, non-symbiotic


Nitrogen is the most abundant element in the atmosphere, however, most often deficient in agricultural lands. This research was an exploratory to get indigenous non-symbiotic nitrogen-fixing bacteria. Soil samples were collectedfrom rhizosphere of green beans. This study was aimed to determine the bacterial population of the three regions; screening, isolation and selection of free nitrogen-fixing bacteria. Antagonism and pathogenicity tests were performed to observe its potential for a biofertilizer product. The highest number of free nitrogen-fixing bacteria was found from forest soil sample of 2.5x 1011CFU/ml. Screening and isolation process has obtained 10 free nitrogen-fixing isolates. Then was selected into 4 isolates namely SNF4, SNF5, SNF7 and SNF8 according to the ammonia production test qualitatively. When an antagonism activity performed, there was no inhibition zone each other. The pathogenicity test did not show the pathogenic symptom. This study also showed that bacterial isolates obtained significantly affected the germination growth of green beans compared to controls. Possibility, bacteria of this type produced growth hormone for a plant. Strain SNF8 has shown the highest ammonium production then was selected for 16S rRNA identification. Similarity test of genome sequence of strain SNF8 had 99% similarity with Bacillus cereus.

Author Biographies

Novi Arfarita, Universitas Islam Malang (UNISMA)

Senior Lecturer

Anton Muhibuddin, Faculty of Agriculture, University of Brawijaya


Tsuyoshi Imai, Graduate School of Science and Engineering, Faculty of Engineering, Yamaguchi University



Arfarita, N., Hidayati, N., Rosyidah, A., Machfudz, M. and Higuchi, T. 2016. Exploration of indigenous soil bacteria producing-exopolysaccharides for stabilizing of aggregates land potential as biofertilizer. Journal of Degraded and Mining Lands Management 4(1): 697-702.

Arfarita, N., Lestari, M.W., Murwani, I. and Higuchi, T. 2017. Isolation of indigenous phosphate solubilizing bacteria from green bean rhizospheres. Journal of Degraded and Mining Lands Management 4(3): 845-851.

Bhardwaj, D., Ansari, M.W., Sahoo, R.K. and Tuteja, N. 2014. Biofertilizers function as key player in sustainable agriculture by improving soil fertility, plant tolerance and crop productivity. Microbial Cell Factories 13:66.

Esteban, K., María, J.M, Patricia, S. and César, J.R. 2010. Organic and inorganic fertilizer effects on a degraded Patagonian rangeland. Plant and Soil 332:135–145.

Fernandez, L., Mercader, J.M., Planas-Felix, M. and Torrents, D. 2014. Adaptation to environmental factors shapes the organization of regulatory regions in microbial communities. BMC Genomics 15(1):877.

Finlay, W.J.J., Logan, N.A. and Sutherland, A.D. 2000. Bacillus cereus produces most emetic toxin at lower temperatures. Letters in Applied Microbiology 31:385–389.

Handayanto, E. and Hairiah, K. 2007. Soil Biology: Platform for Management of Healthy Soil. Pustaka Adipura, Yogyakarta (in Indonesian).

Husen, E., Simanungkalit, R.D.M. and Irawan. 2007. Characterization and quality assessment of Indonesian commercial biofertilizers. Indonesian Journal of Agricultural Science 8: 31-38.

Leghari, S.J., Wahocho, N.A, Laghari, G.M., Talpur, K.H., Wahocho, S.A. and Lashari, A.A. 2016. Role of nitrogen for plant growth and development: a review. Advances in Environmental Biology 10(9): 209-2018.

McGill, W.B. and Cole, C.V. 1981. Comparative aspects of cycling of organic C, N, S and P through soil organic matter. Geoderma 26:267–268. doi: 10.1016/0016-7061(81)90024-0.

Nuraini, Y., Arfarita, N. and Siswanto, B. 2015. Isolation and characteristic of nitrogen-fixing bacteria and phosphate-solubilizing bacteria from soil high in mercury in tailings and compost areas of artisanal gold mine. Agrivita Journal of Agricultural Science 37(1): 1-7.

Parka, M., Kima, C., Yanga, J., Leea, H., Shina. W., Kimb, S. and Sa, T. 2005. Isolation and characterization of diazotrophic growth promoting bacteria from rhizosphere of agricultural crops of Korea. Microbiological Research 160:127—133.

Prihastuti and Harsono, A. 2012. Decrease in the quality of Rhizobium biofertilizers. Sains & Matematika 1(1): 1–5 (in Indonesian).

Roper, M.M. and Gupta V.V.S.R. 2016.Enhancing non-symbiotic N2 fixation in Agriculture. The Open Agriculture Journal 10:7-27.

Azcón, R. and Barea, J.M. 1975. Synthesis of auxins, gibberellins and cytokinins by Azotobacter vinelandii and Azotobacter beijerinckii related to effects produced on tomato plants. Plant and Soil 43(3 ): 609-619.

Subba-Rao, N.S. 1995. Soil Microorganisms and Plant Growth. Science Pub Inc; 3rd edition (December 1995), 350p.

Torsvik, V. and Øvreås, L. 2002. Microbial diversity and function in soil: from genes to ecosystems. Current Opinion in Microbiology 5: 240-245.

West, A.W., Burges, H.D. and Dixon, T.J. 1985. Survival of Bacillus thuringiensis and Bacillus cereus spore inocula in soil: effects of pH, moisture, nutrient availability and indigenous microorganisms. Soil Biology and Biochemistry 17:657–665. doi: 10.1016/0038-0717(85)90043-4.








How to Cite

Arfarita, N., Muhibuddin, A., & Imai, T. (2019). Exploration of indigenous free nitrogen-fixing bacteria from rhizosphere of Vigna radiata for agricultural land treatment. Journal of Degraded and Mining Lands Management, 6(2), 1617–1623. https://doi.org/10.15243/jdmlm.2019.062.1617



Research Article