G- Journal of Environmental Science and Technology 4(6):108-112 (2017) ISSN (Online): 2322-0228 (Print): 2322-021X

G- Journal of Environmental Science and Technology

(An International Peer Reviewed Research Journal)
Available online at http://www.gjestenv.com

The Potentiality of Selected Strain of PGPR: Azotobacter, for Sustainable Agriculture

in India
Ruchi Singh1, Jaspal Singh2, Ravi Deval1, Sudhir Upadhyay3 and Dinesh Kumar1*
1
Department of Biotechnology, Invertis University, Bareilly, U.P. 243123 INDIA
2
Department of Environmental Science, Bareilly College, Bareilly U.P. 243005 INDIA
3
Department of Environmental Science, Purvanchal University, Jaunpur, U.P., INDIA

ARTICLE INFO ABSTRACT

Received: 04 May 2017 In modern agriculture, large numbers of pesticides are being used to inhibit pests to get maximum crop
production .The rapidly growing industrialization along with an increasing population has resulted in the
Revised: 22 July 2017 accumulation of a wide variety of chemicals. One promising treatment method is to exploit the ability of
Accepted: 18 Aug 2017 microorganisms to remove pollutants from contaminated sites. Plant growth promoting rhizobacteria
(PGPR) are a group of free-living bacteria that can be found in the rhizosphere and contribute to enhance
the growth and yield of crop plants. In the last few decades, many microorganisms show positive effect on
Key words: plant development. Azotobacter has been used as a potential nitrogenous fertilizer to increase crop growth.
Azotobacter, Nitrogenous Azotobacter has the capacity of fixing atmospheric nitrogen, it include the production of plant hormones
fertilizer like auxins, gibberellins and cytokinins, nitrogen fixation, These qualities makes Azotobacter the most
successful and widespread group among the PGPR, as well as they produce substances that change the
plant growth and morphology. Co-inoculants of Azotobacter with Rhizobium has been reduced water stress
and dual inoculation of Azospirillium and Azotobacter significantly increased the concentrations of indole-
3-acetic acid (IAA), Potassium, Megnisium, Nitrogen and total soluble sugars (TSS). Azotobacter increases
the tolerance capacity of plants. Azotobacter inoculum with high yielding EPS (exopolysaccharides) and
high nitrogen fixing ability can be utilized to satisfy the future demand of augmented crop production
attributed to increase plant growth promoting agents.

1) INTRODUCTION Azotobacter is an aerobic, free living nitrogen fixer. They

Agricultural sustainability, food security and energy multiply rapidly and develop a thick population in
renewability depends on a healthy and fertile soil. However, rhizosphere, when applied as seed treatment or seedling root-
rapid acceleration of desertification and land degradation by dip or as soil application [5].
numerous anthropogenic activities leads to an estimated loss For sustainable agriculture, the best alternative of chemical
of 24 billion tons of fertile soil from the world’s crop lands fertilizer is necessary because of its adverse effect on the soil
[1]. health. Many alternatives available to enhance the soil fertility
Plant growth promoting rhizobacteria (PGPR) are a group of for better crop production, one promising treatment method is
free-living bacteria that can be found in the rhizosphere and to exploit the ability of microorganisms to remove pollutants
contribute to enhance the growth and yield of crop plants. The from contaminated soils, one of them is Azotobacter. It is an
rhizosphere is the zone that are surrounding by the plant roots important component of integrated nutrient management
in which complex relations occur among the plant, the soil system due to its significant role in soil sustainability [6].
microorganisms and the soil itself. In the last few decades, canal).
many microorganisms show positive effect on plant 1. Taxonomy, Morphology and Distribution of Azotobacter
development. Besides the well-known plant growth promoting The genus Azotobacter belongs to the subclass of the
rhizobacteria, present in the rhizosphere can stimulate the Proteobacteria and comprises seven
plant growth or reduce the damage that are caused by soil- Species: A.chroococcum, A. vinelandii, A. beijerinckii, A.
borne plant pathogens, a process is known as bio-control paspali, A.armeniacus, A. nigricans and A. salinestri [7, 8]
activity [2]. Bacteria identified as PGPR have diverse The taxonomic classification of Azotobacter is shown below:
taxonomy and include strains of the genera Azospirillum, Domain: Bacteria
Azotobacter, Bacillus, Enterobacter, Gordonia, Klebsiella, Kingdom: Bacteria
Paenibacillus, Pseudomonas, Serratia, among others [3]. Phylum: Proteobacteria
Azotobacter has been used as a potential nitrogenous fertilizer Class: Gammproteobacteria
to increase crop growth. The Azotobacter genus was Order: Pseudomonas
discovered in 1901 by Dutch microbiologist and botanist
Beijerinck (founder of environmental microbiology). A
chroococcum is the first aerobic free-living nitrogen fixer [4]. * Corresponding Author: Dr. Dinesh Kumar
Email address: [email protected] 108
Family: Pseudomonadaceae/Azotobaceriaceae fixation. They are extremely tolerant to oxygen while fixing
Genus: Azotobacter nitrogen and this is due to respiration protection of nitrogenase
Azotobacter species are gram negative, free living aerobic [13, 14].
bacteria [9]. The shape of Azotobacter is oval or spherical that Azotobacter species are non-symbiotic heterotrophic bacteria
form thick-walled cysts (means of asexual reproduction under capable of fixing an average 20 kg N/ha/per year [15].
favorable condition) [10].The population of Azotobacter is Bacterization helps to improve plant growth and to increase
generally low in the rhizosphere of the crop plants in soil nitrogen through nitrogen fixation by utilizing carbon for
uncultivated soils. Jensen’s N-free medium is used for rapid its metabolism [16].
multiplication of Azotobacter. Azotobacter grows well at an (b) Role of Azotobacter in growth substances production
optimum temperature range between 20 and 30 °C and grows and promotion
best in neutral to alkaline soil (pH of 6.5-7.5), but does not Azotobacter directly or indirectly affects the plant growth and
develops when the pH is below 6 and hence not present in microbial activity by producing different growth hormone
acidic soil. This rhizobacteria has been reported to present in (IAA and other auxin, such as gibberellins and cytokinins)
the rhizosphere of a number of crop plants such as rice, maize, [17]. Growth substances are natural substances that are
sugarcane, bajra, vegetables and plantation crops [11]. produced by microorganisms. Azotobacter is the genus of
Azotobacter is generally used in any non-legume crop [12]. great interest in agricultural application due to their free
nitrogen fixing ability. Here, emphasis is given on the role of
exo-polysaccharide in sustainable agriculture system and also
to the survival in its own environments [18] Substances like
amino acid produced by Azotobacter are involved in many
processes that explain plant-grown promotion. Biochemical
analysis of chlorophyll, nitrogen, phosphorous, potassium and
protein content was higher in Azotobacter inoculated plants as
compared to non inoculated control plants [19]. Many strains
of Azotobacter also exhibited fungi static properties against
plant pathogens such as Fusarium, Alternaria and
Helminthosporium. The occurrence of this organism has been
reported from the rhizosphere of a number of crop plants such
IOSR Journal of Environmental Science Toxicology and as rice, maize, sugarcane, bajra, vegetables and plantation
Food Technology ,3:67-72 crops, [20].
2. Mode of Action (c) Response of Crops to Growth Promoting Substances
Azotobacter has the capacity of fixing atmospheric nitrogen. Large number of field trials and various experiments carried
Azotobacter include the production of plant hormones like throughout India and whole world has convincingly
auxins, gibberellins and cytokinins, These qualities makes established the importance of Azotobacter as microbial
Azotobacter the most successful and widespread group among inoculant. In India various crops like barley, maize, sugerbeet,
the PGPR, as well as they produce substances that change the carrot, cabbage, potato, wheat, rice, onion, brinjal, tomato and
plant growth and morphology. cabbage were inoculated with Azotobacterin , it increased
(a) Nitrogen fixation germination and growth of seedlings [21].Thus Azotobacter
Azotobacteria is used for nitrogen fixation and inoculation of produced three growth-promoting substances which were
plants due to its rapid growth and high level of nitrogen gibberellin-like in character in that they induced shoot
elongation of dwarf mutant plants whose growth responses are
109
thought to be specific to the gibberellins [22]. Azotobacter regulator that stimulates development of foliage, roots,
produces indole acetic acid (IAA), gibberllic acid (GA) which branching, flowering and fruiting which triggered by fixed
are important plant growth hormones and these hormones will nitrogen [30]. One of the species of Azotobacter vinellandii
helps in seed germination and plant growth considerably [23]. has a potential of salinity stress tolerance in rice. A. vinellandii
Pesticide tolerant Azotobacter species was isolated from paddy sustains growth and improve compatible solutes that results
soils and are known to produce IAA in media supplemented crop improvement.
with 5% pesticide.[24] Dual inoculation of Azotobacter and 5. Effects of Azotobacter on growth and yield of crops
Azospirillum showed synergistic effects by improving growth Azotobacter is urea adaptive bacteria. It is concluded that
prompting hormones, controlling pathogenesis and growth when Azotobacter alone and combination of different conc. of
reducing agents due to producing fungicide antibiotics and urea is applied to rice seeds, the co-inoculation of chemical
compounds (antagonistic effect) and also air molecular and biofertilizers improves the growth of plants [31]. Seedling
Nitrogen fixing and also producing growth prompting of tomato were treated with dual inoculation of Azotobacter
hormones such as auxin, cytokinin and gibberellins and and Azospirillum it showed high performance in whole plant
solving mineral compound [25]. (Plant height, no. of leaves, no. of leaves per plants, no. of
3. Relation to other PGPRs fruits, yield per plants, chlorophyll and protein content). It all
(a) Interaction with Rhizobium treatments co-inoculation shows maximum yield when
Water stress condition has reduced nodule number and nodule compared with single inoculation and control [32].
dry weight. It is observed in many studies that a synergistic The effect of vermicompost and biofertilizers (Azotobacter) on
relation of Azotobacter with Rhizobium interaction as co- the performance of cabbage among the various levels of bio-
inoculants of Azotobacter with Rhizobium has been reduced fertilizers inoculation, Azotobacter resulted in maximum
water stress. Co-inoculants increased most of growth number of leaves/head, while the length of head and head
parameters, water and nutrient uptake under the deficit yield/plant were maximum with Azotobacter/ha. [33].
irrigation because alleviated effect of shortage of water [26].
(b) Interaction with Azospirillum Table: Effect of Azotobacter inoculation on yield of some
The beneficial effects of Azotobacter and Azospirillum
interaction on plants are mainly attributed to improvements in
root development, an increase in the rate of water and mineral
uptake by roots, the displacement of fungi and plant
pathogenic bacteria and to a lesser extent, biological nitrogen
fixation [27].
Some of the studies have shown that a relationship of
Azospirillum and Azotobacter counts in dual inoculations.
These decreases may be due to the antagonistic activity.
Single and dual inoculations stimulated plant growth,
significantly increased the concentrations of indole-3-acetic
acid (IAA), Potassium, Magnesium, Nitrogen and total soluble
sugars (TSS).The significance of IAA, nitrogen fixation and
antibacterial substances, produced by such agronomical
beneficial bacteria [28].
4. Possibility of using Azotobacter in crop Production
Azotobacter has been used as a potential nitrogenous fertilizer important crop plants
to increase crop growth and has the capacity of fixing
atmospheric nitrogen, thus Azotobacter is a best alternative of Average yield in control
Yield
chemical fertilizer. Crop (metric cwt/ha) after
Increase
S Increase in yield over yield incubation
.No. Crops obtained with chemical Barley 21.0 9.1
fertilizers Maize 36.2 8.1
1. Wheat 8-10 Oats 17.2 12.0
2. Rice 5 Potatoes 178.0 8.0
3. Sorghum 15-20 Spring wheat 15.0 8.3
4. Maize 15-20 Sugar beet 283.1 7.2
5. Potato 13 Winter Wheat 21.3 9.8
6. Carrot 16
7. Cauliflower 40 Source -: Microorganism and Plant Growth
8. Tomato 2-24 (Manisha Garg)
9. Cotton 7.24 Azotobacter have shown that the yields of some important
10. Sugarcane 9-24 crop plants can be substantially increased by Azotobacter
inoculation
Bhattacherjee and Dey 2014. [29]
(a) Stress tolerance characteristics 6. CONCLUSION
In the scarcity of water, under adverse condition Azotobacter Azotobacter could be one of the bio-fertilizer options for
increase the tolerance capacity of plants. It produced growth sustainable and environmental eco-friendly. Azotobacter may
110
be further enhanced with the optimization and acclimatization 12)Singh, M.S., Dutta, S. 2006. Mustard and Rapeseed
according to the prevailing soil conditions. In future, they are Response to Azotobacter – A Review. Agric Review,
expected to replace the chemical fertilizers, pesticides and 3:232–234
artificial growth regulators which have numerous side-effects 13)Robson, R.L., Postgate, J.R. 1980 Oxygen and Hydrgen
to sustainable agriculture. Azotobacters are unique Biological Nitrogen Fixation. Ann Review Microbiology,
biofertilizers to maintain the N level in agriculture soil. Co- 34:183–207
inoculants increased most of growth parameters, water and 14)Hakeem, K.R., Akhtar, M.S., Abdullah SNA. 2016. Plant,
nutrient uptake under the deficit irrigation. Azotobacter has a Soil and Microbes-vol.1, Implication in Crop Science
potential to stress tolerance that results crop improvement. Springer International Publishing AG, Gewerbestrasse
Azotobacter shows high performance in whole plant like Plant 11,6330 Cham,Switzerland. 6:366
height, no. of leaves, no. of leaves per plants, no. of fruits, 15)Kizilkaya, R. 2009. Nitrogen Fixation Capacity of
yield per plants, chlorophyll and protein content. Hence, Azotobacter spp. Strains Isolated from Soils in Different
understanding and manipulating this feature may be of great Ecosystems and Relationship between them and The
agro-ecological interest for future crop improvement. Microbiological Properties of Soils. J Environment
Biology, 1: 73-82.
REFERENCE 16)Monib, M., Hosny, I., Fayez, M. 1979a. Effect of
1) FAO 2011. The State of the World’s Land and Water Azotobacter Inoculation on Plant Growth and Soil
Resources for Food and Agriculture (SOLAW)— Nitrogen. Zentralbl Bakteriol Naturwiss, 2:140-148
Managing Systems at Risk. Food and Agriculture 17)Barea, J.M., Brown, M.E. 1974. Effect on Plant growth
Organization of the United Nations, Rome and Earthscan, Produced by A paspali related to the Synthesis of Plant
London. Growth Regulating Substances.J Appl Bacterial ,37: 583-
2) Kloepper, J.W., Schroth, M.N. 1978. Plant Growth 593
Promoting Rhizobacteria on Radishes In Proc Int. Conf. 18)Ishwaq Peerzada, A., Vijai, K., Mohd. Malik, F. 2009.
Plant Pathogen Bacteria 4th Angers, France2: 879–882 Effect of Bio-Organic Fertilizer on the Performance of
3) Glick, B.R. 1995. The Enhancement of Plant Growth by Cabbage under Western U.P. Conditions, Annals of
Free Living Bacteria. Can J Microbiology ,2:109–117 Horticulture, 2(2):204-206
4) Beijerinck, M.W. 1901. Ueber Oligonitophile Mikroben, 19)Kravchenko, L.V., Makarova, N.M., Azarova, T.S. 2002.
Zentralblattfiir Bakteriologie, Parasitenkunde, In Isolation and Phenotypic Characterization of Plant Growth-
Fektionskrankheiten and Hygiene, Abteilung, 7:561-582. Promoting Rhizobacteria with High Anti-phyto pathogenic
5) Maryenko, V.G. 1964. Zavisimost Lurozaja Kukuruzyl Activity and Root-Colonizing Ability. Microbiology
Balansa Azota Azotobacter Chroococcum V Usbvijah ,71:444–448
monobakterialznoj Kultury (Dependence of Maize 20)Elgala, A.M., Ishac, Y.Z., Abdel, M.M., El, Ghandour IAI
Kleyield and nitrogen balance kon Azotobacter 1995. Effect of single and combined inoculation with
Chroococcum in the Jkoodnitions of Monobacterial Azotobacter and VA mycorrhizal fungi on growth and
Lcultivations). Doktl ISHA, 99:399-406. mineral nutrient contents of maize and wheat plants. In:
6) Dev, A. J., Ojha, R.B. and Marahatta, S. 2015. Role of Huang, P. M. et al. (eds.) Environmental Impact of Soil
Azotobacter in Soil Fertility and Sustainability, 6: 6-7. Component Interactions. Metals, Other Inorganics, and
7) Tchan, Y.T. New, P.B. 1984. Genus I Azotobacter, Microbial Activities. Volume II, Lewis Publishers, Boca
Beijerinck 1901, 567AL. In: Krieg, N.R. Holt, J.G. (eds.). Raton, London, Tokyo. 109.
Bergey’s Manual of Determinative Bacteriology, vol. 1, 21)El Shanshoury, A.R. 1995. Interaction of Azotobacter
Williams & Wilkins, Baltimore, USA,1: 220–229. Chroococcum, Azospirillium Brasilense and Streptomyces
8) Becking, J. 2006. The Family Azotobacteraceae. The Butabilis, in Relation to their Effect on Wheat
Prokaryotes. In: Dworkin M, Falkow S, Rosenberg E, Development, 175: 119-127.
Schleifer K-H, Stackebrandt E (eds). Springer New York, 22)Phinney, B.0. and West, C.A. 1960. Gibberellins as Native
pp 759-783 Plant Growth Regulators. A. Review. PI.Physiol, 11: 41, 1.
9) Gandora, V., Gupta, R.D. and Bhardwaj, K.K.R. 1998. 23)Asma, M.T., Pallavi, R.B., Sonal, G.C., Jai, S.G. and
Abundance of Azotobacter in great soil groups of North – Prakash, D.R. 2012. Effect of Endosulfan on Indole Acid
West Himalayas. J. of Indian Society of Soil and Gibberllin Secretion by Azospirillum spp NCIM-2548
Science.3: 379 – 383. and Azotobacter spp NCIM-2452. Int. Res. J. Environment
10)Salhia, M. B. 2010. The Effect of Azotobacter Science. 3:1-4.
Chrococcumas Nitrogen on the Growth and Yield of 24)Chennappa, G., Adkar-Purushothama, C.R., Suraj, U.,
Cucumis sativus. The Islamic University Gaza, Deanery of Tamilvendan, K. and Sreenivasa, M.Y. 2014. Pesticide
Higher Education Faculty of Science, Master of Biological Tolerant Azotobacter Isolates from Paddy Growing Areas
Sciences,Botany. of Northern Karnataka, India. W. J. Microbiol. Biotechnol.
http://library.iugaza.edu.ps/thesis/94335.pdf DOI 10.1007/s11274-013-1412-3,1:1-7
11)Srivastava R. Joshi M, Kumar A. Pachauri Sharma A.K., 25)Naseri, R., Moghadam, A., Darabi, F., Hatami, A.,
Bio-fertilizers for Sustainable Agriculture,(A.K Sharma, Tahmasebei, G.R. 2013. The Effect of Deficit Irrigation
Seema Wahab & Rashmi Srivastava (Eds.) Agriculture and Azotobacter Chroococcum and Azospirillum
Diversification: Problems and Perspectives, pp 58–73) I.K. brasilense on Grain Yield, Yield Components of Maize
International Publishing House Pvt.Ltd. New Delhi. India (S.C.704) as a Second Cropping in Western. Iran Bull
Environ Pharmacology Life Science, 10:104–112

111
26)Dashadi, M., Khosravi, H., Moezzi, A., Habib, N., Heidari,
M. and Radja, R. 2011. Co. Inoculation of Rhizobium and
Azotobacter on Growth Indices of Faba Bean Water Stress
in the Green House Condition, 8: 373-385.
27)Okon, Y., Itzigsohn, R. 1995. The Development of
Azospirillum as a Commercial Inoculants for Improving
Crop Yields. Biotechnology Adv, 13:415–424.
28)Kamil, P., Yami, K.D., Singh, A. 2008. Plant Growth
Promotional Effect of Azotobacter chroococcum,
Piriformospora Indica and Vermicompost on Rice Plant.
Nepal J Science Technology, 9:85–90
29)Bhattacherjee, R., Dey, U. 2014. A Way towards Organic
Farming; A review. Afr J Microbiol Res, 24: 2332–2342.
30)Zena, G.G., Peru, C. 1986. Effect of Different Rates of
Azotobacter and Frequency of Application of Agrispon on
Yield and Quality in the Growing of Onion ( Allium cepa
L.) in Cajamarca. National University of Cajamarca
Faculty of Agriculture Sciences and Forestry S.A. Wani et
al.112:127-133
31)Shrivastava, R., Shrivastava, A.K., Devangan, N. 2015.
Combined Application of Azotobacter and Urea to
Improve Growth of Rice (Oryza Sativum), 3, 67-72.
32)Ramkrishnan, K., Selvkumar, G. 2012. Effect of
Biofertilizers on Enhancement of Growth and Yield on
Tomato (Lyeopersicum esculentum Mill.), 4:20-23.
33)Sharma K, Dak G, Agrawal A, Bhatnagar M, Sharma R.
2007 Effect of phosphate solubilizing bacteria on the
germination of cicer arietinum seeds and seedling growth.
J Herbal Med Toxicol 1, 61–63

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