Plant and Soil

, Volume 377, Issue 1–2, pp 111–126 | Cite as

Alleviation of drought stress in mung bean by strain Pseudomonas aeruginosa GGRJ21

Regular Article


Backgrounds and aims

Mung bean (Vigna radiata (L.) R. Wilczek), a widely cultivated pulse crops in India, experiences severe drought stress during the cultivation period. Apart from the conventional plant breeding and transgenic approaches, the application of plant growth-promoting rhizobacteria (PGPR) has always been a promising approach to improve abiotic stress tolerance in crop plants. The aim of the present study was to investigate the role of mung bean rhizosphere-associated Pseudomonas aeruginosa GGRJ21 strain on drought stress alleviation in the host plant.


Fluorescent pseudomonads were isolated from mung bean rhizosphere by employing a culture-dependent approach. The role of osmotic stress tolerant P. aeruginosa GGRJ21 on drought stress alleviation in host plants was further examined in both the green house and field conditions.


An elevated production of reactive oxygen species scavenging enzymes and cellular osmolytes; increased root length, shoot length, dry weight, relative water content; and a stronger upregulation of three drought stress-responsive genes, i.e., dehydration-responsive element binding protein (DREB2A), catalase (CAT1), and dehydrin (DHN) were observed in GGRJ21 inoculated plants in comparison with the uninoculated control plants tested under drought conditions. The field experimental data show an increase in biomass and better growth and development in inoculated and stressed plants when compared with untreated and stressed plants.


P. aeruginosa GGRJ21 strain was found to elicit water stress tolerance in mung bean plants by accelerating the accumulation of inherent levels of antioxidant enzymes, cell osmolytes, and consistently expediting the upregulation of stress responsive genes in PGPR-treated plants under water stress conditions.


Drought stress Pseudomonas aeruginosa Catalase Peroxidase Superoxide dismutase Proline Reactive oxygen species 



The work is supported by a Network Project (AMAAS), sponsored by the Indian Council of Agricultural Research (ICAR), the Government of India, New Delhi. The authors are thankful to the Director, CSIR-NEIST, and the Head, Biotechnology Division, CSIR-NEIST, for providing necessary facilities to carry out the work. The authors are also thankful to the MAEP Division, CSIR-NEIST for providing necessary area in experimental farm to conduct the field experiment.

Supplementary material

11104_2013_1981_MOESM1_ESM.docx (14 kb)
Table S1 (DOCX 13 kb)
11104_2013_1981_MOESM2_ESM.docx (17 kb)
Table S2 (DOCX 16 kb)
11104_2013_1981_MOESM3_ESM.docx (24 kb)
Fig. S1 (DOCX 23 kb)


  1. Alexander DB, Zuberer DA (1991) Use of chrome azurol S reagents to evaluate siderophore production by rhizosphere bacteria. Biol Fertil Soils 12:39–45CrossRefGoogle Scholar
  2. Ashraf M, Akram NA (2009) Improving salinity tolerance of plants through conventional breeding and genetic engineering: an analytical comparison. Biotechnol Adv 27:744–752PubMedCrossRefGoogle Scholar
  3. Baki AAA, Anderson JD (1973) Vigour determination in soybean seed by multiple criteria. Crop Sci 13:630–633CrossRefGoogle Scholar
  4. Balcht A, Smith R (1994) Pseudomonas aeruginosa: infections and treatment. Informa Health Care, UK, ISBN 0-8247-9210-6, pp. 83–84Google Scholar
  5. Bashan Y, de-Bashan LE (2005) Bacteria/plant growth-promotion. In: Hillel D (ed) Encyclopedia of soils in the environment, vol 1. Elsevier Oxford, UK, pp 103–115Google Scholar
  6. Bashan Y, Holguin G, de-Bashan LE (2004) Azospirillum-plant relationships: physiological, molecular, agricultural, and environmental advances (1997–2003). Can J Microbiol 50:521–577PubMedCrossRefGoogle Scholar
  7. Bashan Y, Kamnev AK, de-Bashan LE (2013) Tricalcium phosphate is inappropriate as a universal selection factor for isolating and testing phosphate-solubilizing bacteria that enhance plant growth: a proposal for an alternative procedure. Biol Fertil Soils 49:465–479CrossRefGoogle Scholar
  8. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  9. Berg G, Eberl L, Hartmann A (2005) The rhizosphere as a reservoir for opportunistic human pathogenic bacteria. Env Microbio l 7:1673–1685CrossRefGoogle Scholar
  10. Beyer JWF, Fridovich I (1987) Assaying for super oxide dismutase activity: some large consequences of minor changes in conditions. Anal Biochem 161:559–566PubMedCrossRefGoogle Scholar
  11. Bowen GD, Rovira AD (1991) The rhizosphere, the hidden half of the hidden half. In: Waisel Y, Eshel A, Kafkafi U (eds) Plant roots, the hidden half. Marcel Dekker, Inc, New York, USA, pp 641–629Google Scholar
  12. Bradford MM (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  13. Chowdhury SP, Schmid M, Hartmann A, Tripathi AK (2007) Identification of diazotrophs in the culturable bacterial community associated with roots of Lasiurus sindicus, a perennial grass of Thar Desert, India. Microb Ecol 54:82–90PubMedCrossRefGoogle Scholar
  14. D’Souza-Ault MR, Smith LT, Smith GM (1993) Roles of N acetylglutaminylglutamine amide and glycine betaine in adaptation of Pseudomonas aeruginosa to osmotic stress. Appl Env Microbiol 59:473–478Google Scholar
  15. de Zelicourt A, Al-Yousif M, Hirt H (2013) Rhizosphere microbes as essential partners for plant stress tolerance. Mol Plant 6:242–245PubMedCrossRefGoogle Scholar
  16. de-Bashan LE, Hernandez JP, Bashan Y (2012) The potential contribution of plant growth-promoting bacteria to reduce environmental degradation—a comprehensive evaluation. Appl Soil Ecol 61:171–189CrossRefGoogle Scholar
  17. Dworkin M, Foster J (1958) Experiments with some microorganisms which utilize ethane and hydrogen. J Bacteriol 75:592–601PubMedCentralPubMedGoogle Scholar
  18. Edwards U, Rogall TH, Blocker H, Emde M, Bottger EC (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nuc Acids Res 17:7843–7853CrossRefGoogle Scholar
  19. Egamberdieva D (2012) The management of soil quality and plant productivity in stressed environment with rhizobacteria. In: Maheshwari DK (ed) Bacteria in agrobiology: stress management. Springer-Verlag Berlin Heidelberg, Germany, pp 27–40CrossRefGoogle Scholar
  20. Egamberdieva D, Kamilova F, Validov S, Gafurova L, Kucharova Z, Lugtenberg B (2008) High incidence of plant growth-stimulating bacteria associated with the rhizosphere of wheat grown on salinated soil in Uzbekistan. Env Microbiol 10:1–9Google Scholar
  21. Farooq M, Wahid A, Kobayashi N, Fujita D, Basra SMA (2009) Plant drought stress: effects, mechanisms and management. Agron Sustain Dev 29:185–212CrossRefGoogle Scholar
  22. Figueiredo MVB, Burity HA, Martinez CR, Chanway CP (2008) Alleviation of drought stress in common bean (Phaseolus vulgaris L.) by co-inoculation with Paenibacillus polymyxa and Rhizobium tropici. Appl Soil Ecol 40:182–188CrossRefGoogle Scholar
  23. Fiske CH, Subbarow Y (1925) A colorimetric determination of phosphorus. J Biol Chem 66:375–400Google Scholar
  24. Gilbert GS, Parke JL, Clayton MK, Handelsman J (1993) Effects of an introduced bacterium on bacterial communities on roots. Ecology 74:840–854CrossRefGoogle Scholar
  25. Glick BR (2005) Modulation of plant ethylene levels by the bacterial enzyme ACC deaminase. FEMS Microbiol Lett 251:1–7PubMedCrossRefGoogle Scholar
  26. Glick BR, Todorovic B, Czarny J, Cheng Z, Duan J, McConkey B (2007) Promotion of plant growth by bacterial ACC deaminase. Crit Rev Plant Sci 26:227–242CrossRefGoogle Scholar
  27. Gordon SA, Weber RP (1951) Colorimetric estimation of indole acetic acid. Plant Physiol 26:192–195PubMedCentralPubMedCrossRefGoogle Scholar
  28. Grieve CM, Grattan SR (1983) Rapid assay for determination of water soluble quaternary ammonium compounds. Plant Soil 70:303–307CrossRefGoogle Scholar
  29. Gururani MA, Upadhyaya CP, Baskar V, VenkateshJelli, Nookaraju A, Park SW (2013) Plant growth-promoting rhizobacteria enhance abiotic stress tolerance in Solanum tuberosum through inducing changes in the expression of ROS-scavenging enzymes and improved photosynthetic performance. J Plant Growth Regul 32:245–258CrossRefGoogle Scholar
  30. Hayat R, Ali S, Amara U, Khalid R, Ahmed I (2010) Soil beneficial bacteria and their role in plant growth promotion: a review. Ann Microbiol 60:579–598CrossRefGoogle Scholar
  31. Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Circle 347. University of California Agricultural Experimental station Station, BerkleyGoogle Scholar
  32. Honma M, Shimomura T (1978) Metabolism of 1-aminocyclopropane-1-carboxylic acid. Agri Biol Chem 42:1825–1831CrossRefGoogle Scholar
  33. ISTA (1993) Proceedings of the International Seed Testing Association, international rules for seed testing. Seed Sci Technol 21:25–30Google Scholar
  34. Jablasone J, Warrinera K, Griffithsa M (2005) Interactions of Escherichia coli O157:H7, Salmonella typhimurium and Listeria monocytogenes plants cultivated in a gnotobiotic system. Int J Food Microbiol 99:7–18PubMedCrossRefGoogle Scholar
  35. Ji P, Wilson M (2002) Assessment of the importance of similarity in carbon source utilization profiles between the biological control agent and the pathogen in biological control of bacterial speck of tomato. App Environ Microbiol 68:4383–4389CrossRefGoogle Scholar
  36. Kar M, Mishra D (1976) Catalase, peroxidase, and polyphenoloxidase activities during rice leaf senescence. Plant Physiol 57:315–319PubMedCentralPubMedCrossRefGoogle Scholar
  37. Kim Y-C, Glick BR, Bashan Y, Ryu CM (2012) Enhancement of plant drought tolerance by microbes. In: Aroca R (ed) Plant responses to drought stress: from morphological to molecular features. Springer Verlag, Berlin & Heidelberg, Germany pp, pp 383–413CrossRefGoogle Scholar
  38. Kohler J, Hernandez JA, Caravaca F, Roldàn A (2008) Plant-growth-promoting rhizobacteria and abuscularmycorrhizal fungi modify alleviation biochemical mechanisms in water-stressed plants. Funct Plant Biol 35:141–151CrossRefGoogle Scholar
  39. Kremer RJ, Souissi T (2001) Cyanide production by rhizobacteria and potential for suppression of weed seedling growth. Curr Microbiol 43:182–186PubMedCrossRefGoogle Scholar
  40. Krulwich TA, Sachs G, Padan E (2011) Molecular aspects of bacterial pH sensing and homeostasis. Nat Rev Microbiol 9:330–343PubMedCentralPubMedCrossRefGoogle Scholar
  41. Liddycoat SM, Greenberg BM, Wolyn DJ (2009) The effect of plant growth-promoting rhizobacteria an asparagus seedling and germinating seeds subjected to water stress under greenhouse conditions. Can J Microbiol 55:388–394PubMedCrossRefGoogle Scholar
  42. Lorenzen CJ (1967) Determination of chlorophyll and phaeopigments: spectrophotometric equations. ASLO: Limnol Oceano 12:343–346Google Scholar
  43. Mayak S, Tirosh T, Glick BR (2004) Plant growth-promoting bacteria that confer resistance to water stress in tomatoes and peppers. Plant Sci 166:525–530CrossRefGoogle Scholar
  44. Michel BE, Kaufmann MR (1973) The osmotic potential of polyethylene glycol 6000. Plant Physiol 51:914–916PubMedCentralPubMedCrossRefGoogle Scholar
  45. Myers N, Mittermeir RA, Mittermeir CG, da Fonseca GAB, Kents J (2000) Biodiversity hotspots for conservation priorities. Nature 403:853–858PubMedCrossRefGoogle Scholar
  46. Nakashima K, Shinwari ZK, Sakuma Y, Seki M, Miura S, Shinozaki K, Yamaguchi-Shinozaki K (2000) Organization and expression of two Arabidopsis DREB2 genes encoding DRE-binding proteins involved in dehydration- and high-salinity-responsive gene expression. Plant Mol Biol 42:657–665PubMedCrossRefGoogle Scholar
  47. Nakkeeran S, Fernando WGD, Siddiqui ZA (2005) Plant growth promoting rhizobacteria formulations and its scope in commercialization for the management of pests and diseases. In: Siddiqui ZA (ed) PGPR: biocontrol and biofertilization. Springer, Dordrecht, The Netherlands, pp 257–296, ISBN 978-1-4020-4152-5Google Scholar
  48. Ochman H, Lawrence JG, Groisman EA (2000) Lateral gene transfer and the nature of bacterial innovation. Nature 405:299–304PubMedCrossRefGoogle Scholar
  49. Oliver A, Canton R, Campo P, Baquero F, Blazquez J (2000) High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science 288:1251–1254PubMedCrossRefGoogle Scholar
  50. Patten CL, Glick BR (2002) Role of Pseudomonas putida indole acetic acid in development of host plant root system. Appl Environ Microbiol 48:3795–3801CrossRefGoogle Scholar
  51. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):e45PubMedCentralPubMedCrossRefGoogle Scholar
  52. Rahman RN, Geok LP, Basri M, Salleh AB (2005) Physical factors affecting the production of organic solvent-tolerant protease by Pseudomonas aeruginosa strain K. Bioresour Technol 96:429–436PubMedCrossRefGoogle Scholar
  53. Ramos JL, Duque E, Gallegos MT, Godoy P, Ramos-Gonzalez MI, Rojas A, Teran W, Segura A (2002) Mechanisms of solvent tolerance in gram-negative bacteria. Annu Rev Microbiol 56:743–768PubMedCrossRefGoogle Scholar
  54. Reeves M, Pine L, Neilands JB, Bullows A (1983) Absence of siderophore activity in Legionella sp. grown in iron deficient media. J Bacteriol 154:324–329PubMedCentralPubMedGoogle Scholar
  55. Roberson EB, Firestone MK (1992) Relationship between desiccation and exopolysaccharide production in soil Pseudomonas sp. Appl Environ Microbiol 58:1284–1291PubMedCentralPubMedGoogle Scholar
  56. Roberts DP, Dery PD, Yucel I, Buyer JS (2000) Importance of pfk A for rapid growth of Enterobacter cloacae during colonization of crop seed. Appl Environ Microbiol 66:87–91PubMedCentralPubMedCrossRefGoogle Scholar
  57. Sandhya V, Ali SKZ, Grover M, Reddy G, Venkateswarlu B (2009) Alleviation of drought stress effects in sunflower seedlings by the exopolysaccharides producing Pseudomonas putida strain GAP-P45. Biol Fert Soils 46:17–26CrossRefGoogle Scholar
  58. Saravanakumar D, Kavino M, Raguchander T, Subbian P, Samiyappan R (2011) Plant growth promoting bacteria enhance water stress resistance in green gram plants. Acta Physiol Plant 33:203–209CrossRefGoogle Scholar
  59. Sardessai Y, Bhosle S (2002) Tolerance of bacteria to organic solvents. Res Microbiol 153:263–268PubMedCrossRefGoogle Scholar
  60. Sardessai YN, Bhosle S (2004) Industrial potential of organic solvent tolerant bacteria. Biotechnol Prog 20:655–660PubMedCrossRefGoogle Scholar
  61. Scandalios JG (1994) Regulation and properties of plant catalases. In: Foyer CH, Mullineaux PM (eds) Causes of photooxidative stress and amelioration of defense systems in plants. CRC Press, Boca Raton, Florida, pp 275–315Google Scholar
  62. Shahjee HM, Banerjee K, Ahmad F (2002) Comparative analysis of naturally occurring l-amino acid osmolytes and their d-isomers on protection of Escherichia coli against environmental stresses. J Biosci 27:515–520PubMedCrossRefGoogle Scholar
  63. Sikkema J, de Bont JA, Poolman B (1995) Mechanisms of membrane toxicity of hydrocarbons. Microbiol Rev 59:201–222PubMedCentralPubMedGoogle Scholar
  64. Thapa G, Dey M, Sahoo L, Panda SK (2011) An insight into the drought stress induced alterations in plants. Biol Plant 55:603–613CrossRefGoogle Scholar
  65. Tonon G, Kevers C, Faivre-Rampant O, Graziani M, Gaspar T (2004) Effect of NaCl and mannitoliso-osmotic stresses on proline and free polyamine levels in embryogenic Fraxinus angustifolia callus. J Plant Physiol 161:701–708PubMedCrossRefGoogle Scholar
  66. USDA (2010) Soil taxonomy. A basic system of soil classification for making and interpreting soil surveys. United States Department of Agriculture, Washington, DCGoogle Scholar
  67. Van Loon LC, Bakker PAHM, Pieterse CMJ (1998) Systematic resistance induced by rhizosphere bacteria. Annu Rev Pathol 36:453–483Google Scholar
  68. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, de Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:research0034-research0034.11.doi: 10.1186/gb-2002-3-7-research0034
  69. Weatherly PE (1950) Studies in the water relations of the cotton plant I. The field measurement of water deficit in leaves. New Phytol 49(1):81–97CrossRefGoogle Scholar
  70. Wilkening S, Bader A (2004) Quantitative real-time polymerase chain reaction: methodical analysis and mathematical model. J Biomol Tech 15:107–111PubMedCentralPubMedGoogle Scholar
  71. Yuwono T, Handayani D, Soedarsono J (2005) The role of osmotolerant rhizobacteria in rice growth under different drought conditions. Aus J Agri Res 56:715–721CrossRefGoogle Scholar
  72. Ziska LH (2011) Climate change, carbon dioxide and global crop production: food security and uncertainty. In: Dinar A, Mendelsohn R (eds) Handbook on climate change and agriculture. Edward Elgar Publishing Ltd, Cheltenham and Camberley, UK, pp 9–31Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.Biotechnology DivisionCSIR-North East Institute of Science and TechnologyJorhatIndia

Personalised recommendations