Isolation of Indigenous Azotobacter from the Soil of Different Regions of Tehran and Investigating the Effect of Their Inoculation on Tomato (Solanum lycopersicum L.) Plant Growth

Subject Areas : Journal of Crop Ecophysiology

Shaghayegh Golchin Irani ¹ , Gholamreza Taheri Sangsari ² , Akram Sadat Tabatabaee Bafroee ³ , Mohammad Javad Avesta ⁴

1 - MS.c. Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran
2 - Faculty Member, Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran
3 - Assistant Professor, Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran
4 - Member of Royan Tisan Sabz Knowledge-Based Company, Tehran, Iran

Received: 2020-10-06 Accepted : 2021-08-22 Published : 2022-09-23

Keywords: Optimization, Biological fertilizer, Azotobacter, isolation, nif H, Tomato plant,

Abstract :

Azotobacter is an aerobic, gram negative and chemoorganotrophic bacterium, that is able to stabilize molecular nitrogen nonsymbiotically. The role of Azotobacter in plant growth is due to the production of growth-promoting hormones, the ability to dissolve insoluble phosphates, nitrogen fixation, increase stress resistance and biocontrol of plant pathogens. The aim of this study was to isolate and identify the indigenous Azotobacter from the soil of different areas of Tehran. The effect of tomato plant inoculation with isolates on growth promoting was also investigated. Finally, the growth conditions of the superior isolate were optimized.  Azotobacter isolates were obtained from soil samples using serial dilution method and identified by conventional biochemical tests. The nif H gene, encoding nitrogenase enzyme, was identified in isolates using real-time PCR technique. Then the tomato seeds were inoculated with isolates and seedling growth rate including stem and root length were measured during 34 days. The parameters of temperature, pH, aeration rate, and carbon and nitrogen sources were optimized for superior isolate. In this study, 27 isolates were identified as nitrogen fixing Azotobacter. Considering the results, all isolates showed a significant increase (p <0.05) in stem and root length of tomato plants compared to standard strain and negative control. Among them, isolate No. 21 had the greatest effect during 34 days of study and its best growth conditions in the presence of mannitol carbon source, peptone nitrogen source, 200 rpm rotation, 30°C and pH 7were acquired. According to the results of this study, the obtained indigenous isolates particularly isolate No.21 have the potential to be used as biological fertilizer after further investigations.

References:

• Aasfar, A., A. Bargaz, K. Yaakoubi, A. Hilali, I. Bennis, Y. Zeroual, and I. Meftah Kadmiri. 2021. Nitrogen fixing azotobacter species as potential soil biological enhancers for crop nutrition and yield stability. Frontiers in Microbiology. 12:


• Alalaf, A. H. 2020. The role of biofertilization in improving fruit productivity: a review. International Journal of Agricultural and Statistical Sciences. 16: 107-112.


• Ardakani, M.R., D. Mazaheri, and G. Nourmohammadi. 2001. Effect of azospirillum, mycorrhiza and streptomyces with manure utilization on yield and yield component of wheat (mahdavi var.). Journal of Agriculturan Science. 7: 1- 16.


• Arora, M., P. Saxena, M.Z. Abdin, and A. Varma. 2018. Interaction between Piriformospora indica and Azotobacter chroococcum governs better plant physiological and biochemical parameters in Artemisia annua plants grown under in vitro conditions. Symbiosis. 75: 103–112.


• Aseri, G.K., N. Jain, J. Panwar, A.V. Rao, and P.R. Meghwal. 2008. Biofertilizers improve plant growth, fruit yield, nutrition, metabolism and rhizosphere enzyme activities of pomegranate (Punica granatum) in Indian Thar Desert. Scientia Horticulturae. 117: 130–135.


• Chavada, N.B., R. Patel, S. Vanpuria, B.P. Raval, and P.V. Thakkar. 2010. A study on isolated diazotrophic (non-symbiotics) bacteria from saline desert soil as a biofertilizer. International Journal of Pharmaceutical Sciences and Researches. 1: 52–54.


• Dadok, M., M. Beglarian, S. Mehrabian, H. Zali, M. Zamanian azodi, and M. Salehi. 2013. Phylogenetic identification of nitrogen-fixing bacteria isolated from the rhizosphere of asparagus plants using 16s rRNA and the effect of zinc on isolated strains. Scientific Journal of Ilam University of Medical Sciences. 20(5): 112–20.


• Dadok, M., Mehrabian, S., Salehi, M., and Irian, S. 2014. Morphological, biochemical and molecular characterization of twelve nitrogen-fixing bacteria and their response to various zinc concentration. Jundishapur Journal of Microbiology, 7:


• Das, K., R. Dang, and T.N. Shivananda. 2008. Influence of bio-fertilizers on the availability of nutrients (N, P and K) in soil in relation to growth and yield of Stevia rebaudiana grown in South India. International Journal of Applied Research in Natural Products. 1: 20-24.


• Dupin, S.E., R. Geurts, and E.T. Kiers. 2020. The non-legume Parasponia andersonii mediates the fitness of nitrogen-fixing rhizobial symbionts under high nitrogen conditions. Frontiers in Plant Science. 10: 1779-1789.


• El-Zeiny, O.A.H. 2007. Effect of biofertilizers and root exudates of two weed as a source of natural growth regulators on growth and productivity of bean plants (Phaseolus vulgaris). Journal of Agricultural and Biological Science. 3: 440–446.


• Esbati, M., A. Akhavan Sepahi, A. Asgharzadeh, and M. Khosrow Shahli. 2014. Isolation, identification and population study of Azospirillum In soils around Tehran and evaluation of their growth stimulant effects on tomato plants under greenhouse conditions. Soil Biology. 2(1): 43-54. (In Persian).


• Haghighi, S., T.S. Nejad, and S. Lack. 2011. Calculate the growth dynamics of root and shoot of bean plants. Journal of American Science. 7: 19–26.


• Hajeeboland, R., N. Asgharzadeh, and Z. Mehrfar. 2004. Ecological study of Azotobacter in two pasture lands of the north-west Iran and its inoculation effect on growth and mineral nutrition of wheat (Triticum aestivum cv. Omid) plants. JWSS-Isfahan University of Technology. 8: 75–90. (In Persian).


• Hasanudin, H. 2003. Increasing of the nutrient and uptake avaliability of N and P and through corn yield of inoculation of mycorrhiza, azotobacter and on ultisol organic matter. Journal of Agriculture Sciences of Indonesia. 5: 83–89.


• Khosravi, H., and H. Mohammadi. 2013. Investigation of the effects of inoculation of Tobacteria with fertilizer on dryland wheat. Journal of Soil Management and Sustainable Production. 3(2): 219-205. (In Persian).


• Kumar, V., R.K. Behl, and N. Narula. 2001. Establishment of phosphate-solubilizing strains of Azotobacter chroococcum in the rhizosphere and their effect on wheat cultivars under green house conditions. Microbiological Research. 156: 87–93.


• Kumar, V., and K.P. Singh. 2001. Enriching vermicompost by nitrogen fixing and phosphate solubilizing bacteria. Bioresource Technology. 76: 173–175.


• Kumar, G.P., S.K. Yadav, P.R. Thawale, S.K. Singh, and A.A. Juwarkar. 2008. Growth of Jatropha curcas on heavy metal contaminated soil amended with industrial wastes and Azotobacter –A greenhouse study. Bioresource Technology. 99: 2078–2082.


• Kurdish, I.K., Z.T. Bega, and I.Y. Tsarenko. 2006. The effects of several factors on the growth of pure and mixed cultures of Azotobacter chroococcum and Bacillus subtilis. Applied Biochemistry and Microbiology. 42: 278–283.


• Martin, X.M., C.S. Sumathi, and V.R. Kannan. 2011. Influence of agrochemicals and Azotobacter sp. application on soil fertility in relation to maize growth under nursery conditions. Eurasian Journal of Biosciences. 5: 19–28.


• Mukhtar, H., H. Bashir, A. Nawaz, and I. Haq. 2018. Optimization of growth conditions for Azotobacter species and their use as biofertilizer. Jounnal of Bacteriology and Mycology. 6: 274-278.


• Nosheen, A., A. Bano, and F. Ullah. 2016. Bioinoculants: a sustainable approach to maximize the yield of Ethiopian mustard (Brassica carinata) under low input of chemical fertilizers. Toxicology and Industrial Health. 32: 270–277.


• Rajaee, S., H.A. Alikhani, and F. Raiesi. 2007. Effect of plant growth promoting potentials of Azotobacter chroococcum native strains on growth, yield and uptake of nutrients in wheat. Journal of Crop Production and Processing. 11: 285–297. (In Persian).


• Soleimanifard, A., M. Mojaddam, S. Lack, and M. Alavifazel. 2022. Effect of azotobacter and nitrogen fertilizer levels on agro-physiological traits and yield of safflower (Carthamus tinctorius) genotypes under different moisture conditions. Journal of Crop Ecophysiology. 15: 467-492.


• Soumare, A., A.G. Diedhiou, M. Thuita, and M. Hafidi. 2020. Exploiting biological nitrogen fixation: a route towards a sustainable agriculture. Plants. 9: 1011-1033.


• Romero-Perdomo, F., J. Abril, M. Camelo, A. Moreno-Galván, I. Pastrana, D. Rojas-Tapias, and R. Bonilla. 2017. Azotobacter chroococcum as a potentially useful bacterial biofertilizer for cotton (Gossypium hirsutum): Effect in reducing N fertilization. Revista Argentina de Microbiologia. 49: 377-383.


• Tran, Q., D.T. Pham, and V. Phan. 2017. Using 16S rRNA gene as marker to detect unknown bacteria in microbial communities. BMC Bioinformatics. 18: 155-161.


• Yasari, E., M.A.E. Azadgoleh, S. Mozafari, and M.R. Alashti. 2009. Enhancement of growth and nutrient uptake of rapeseed (Brassica napus) by applying mineral nutrients and biofertilizers. Pakistan Journal of Biological Sciences. 12: 127–133.