Plant and Soil

, Volume 366, Issue 1–2, pp 93–105 | Cite as

Isolation of ACC deaminase producing PGPR from rice rhizosphere and evaluating their plant growth promoting activity under salt stress

  • Himadri Bhusan Bal
  • Lipika Nayak
  • Subhasis Das
  • Tapan K. Adhya
Regular Article



Bacteria possessing ACC deaminase activity reduce the level of stress ethylene conferring resistance and stimulating growth of plants under various biotic and abiotic stresses. The present study aims at isolating efficient ACC deaminase producing PGPR strains from the rhizosphere of rice plants grown in coastal saline soils and quantifying the effect of potent PGPR isolates on rice seed germination and seedling growth under salinity stress and ethylene production from rice seedlings inoculated with ACC deaminase containing PGPR.


Soils from root region of rice growing in coastal soils of varying salinity were used for isolating ACC deaminase producing bacteria and three bacterial isolates were identified following polyphasic taxonomy. Seed germination, root growth and stress ethylene production in rice seedlings following inoculation with selected PGPR under salt stress were quantified.


Inoculation with selected PGPR isolates had considerable positive impacts on different growth parameters of rice including germination percentage, shoot and root growth and chlorophyll content as compared to uninoculated control. Inoculation with the ACC deaminase producing strains reduced ethylene production under salinity stress.


This study demonstrates the effectiveness of rhizobacteria containing ACC deaminase for enhancing salt tolerance and consequently improving the growth of rice plants under salt-stress conditions.


Growth promoting rhizobacteria Coastal rice soils ACC deaminase Polyphasic taxonomy Salinity stress Ethylene production 



This work was supported in part by the ICAR Networking Project, “Application of Microorganisms in Agriculture and Allied Sciences (AMAAS) - theme Microbial Diversity and Identification” by the Indian Council of Agricultural Research, New Delhi.

Supplementary material

11104_2012_1402_MOESM1_ESM.doc (56 kb)
Supplementary Table 1 Carbon source utilization profile of the isolates using the BIOLOG GEN III MicroPlates (DOC 55 kb)
11104_2012_1402_MOESM2_ESM.doc (61 kb)
Supplementary Table 2 Percentage of total fatty acids present in isolates (DOC 61 kb)


  1. Asch F, Dingkuhn M, Dorffling K (2000) Salinity increases CO2 assimilation but reduces growth in field grown, irrigated rice. Plant Soil 218:1–10CrossRefGoogle Scholar
  2. Belimov AA, Hontzeas N, Safronova VI, Demchinskaya SV, Piluzza G, Bullitta S, Glick BR (2005) Cadmium-tolerant plant growth-promoting rhizobacteria associated with the roots of Indian mustard (Brassica juncea L. Czern). Soil Biol Biochem 37:241–250CrossRefGoogle Scholar
  3. Belimov AA, Safronova VI, Mimura T (2002) Response of spring rape (Brassica napus L. var. Oleifera) to inoculation with plant growth promoting rhizobacteria containing 1-aminocyclopropane-1-carboxylate deaminase depends on nutrient status of the plant. Can J Microbiol 48:189–199PubMedCrossRefGoogle Scholar
  4. Bharathi R, Vivekananthan R, Harish S, Ramanathan A, Samiyappan R (2004) Rhizobacteria-based bio-formulations for the management of fruit rot infection in chillies. Crop Prot 23:835–843CrossRefGoogle Scholar
  5. Corsini A (2011) Microbial arsenic transformations in contaminated soils. PhD thesis, Universita’ degli Studi di MilanoGoogle Scholar
  6. Dimkpa C, Weinan T, Asch F (2009) Plant–rhizobacteria interactions alleviate abiotic stress conditions. Plant Cell Environ 32:1682–1694PubMedCrossRefGoogle Scholar
  7. Dworkin M, Foster J (1958) Experiments with some microorganisms which utilize ethane and hydrogen. J Bacteriol 75:592–601PubMedGoogle Scholar
  8. Fišerová H, Mikušová Z, Klemš M (2008) Estimation of ethylene production and 1-aminocyclopropane-1-carboxylic acid content in plants by means of gas chromatography. Plant Soil Environ 54:55–60Google Scholar
  9. Gholami A, Shahsavani S, Nezarat S (2009) The effect of Plant Growth Promoting Rhizobacteria (PGPR) on germination, seedling growth and yield of maize. Int J Biol Life Sci 5:35–40Google Scholar
  10. Glick BR (1995) The enhancement of plant growth by free-living bacteria. Can J Microbiol 41:109–117CrossRefGoogle Scholar
  11. Glick BR (2004) Bacterial ACC deaminase and the alleviation of plant stress. Adv Appl Microbiol 56:291–312PubMedCrossRefGoogle Scholar
  12. Glick BR (2005) Modulation of plant ethylene levels by the enzyme ACC deaminase. FEMS Microbiol Lett 251:1–7PubMedCrossRefGoogle Scholar
  13. Glick BR, Penrose DM, Li J (1998) A model for the lowering of plant ethylene concentration by plant growth-promoting bacteria. J Theor Biol 190:63–68PubMedCrossRefGoogle Scholar
  14. Glick BR, Cheng Z, Czarny J, Duan J (2007a) Promotion of plant growth by ACC deaminase-containing soil bacteria. Eur J Plant Pathol 119:329–339CrossRefGoogle Scholar
  15. Glick BR, Todorovic B, Czarny J, Cheng Z, Duan J, McConkey B (2007b) Promotion of plant growth by bacterial ACC deaminase. Crit Rev Plant Sci 26:227–242CrossRefGoogle Scholar
  16. Glickmann E, Dessaux Y (1995) A critical examination of the specificity of the Salkowski reagent for indolic compounds produced by phytopathogenic bacteria. Appl Environ Microbiol 61:793–796PubMedGoogle Scholar
  17. Gomez KA, Gomez AA (1984) Statistical procedures for agricultural research, 2nd edn. John Wiley and Sons Inc, SingaporeGoogle Scholar
  18. Hazra C, Kundu D, Ghosh P, Joshi S, Dandi N, Chaudhari A (2011) Screening and identification of Pseudomonas aeruginosa AB4 for improved production, characterization and application of a glycolipid biosurfactant using low-cost agro-based raw materials. J Chem Technol Biotechnol 86:185–198CrossRefGoogle Scholar
  19. Hoagland DR, Boyer, TC (1936) General nature of the process of salt accumulation by roots with description of experimental method. Plant Physiol 11:471–507Google Scholar
  20. Holt JG, Kreig NR, Sneath PHA, Staley JT, Williams ST (1994) Bergey’s Manual of determinative bacteriology, 9th edn. Williams and Wilkins, BaltimoreGoogle Scholar
  21. Islam MR, Madhaiyan M, Deka Boruah HP, Yim W, Lee G, Saravanan VS, Fu Q, Hu H, Sa T (2009) Characterization of plant growth-promoting traits of three-living diazotrophic bacteria and their inoculation effects on growth and nitrogen uptake of crop plants. J Microbiol Biotech 19:1213–1222Google Scholar
  22. Ismail AM, Thapa B, Egdane J (2009) Salinity tolerance in rice: physiological bases and implications on management strategies for better crop establishment. In: Improving productivity and livelihood for fragile environments. IRRI technical bull. 13. International Rice Research Institute, Los Banos, pp 8–13Google Scholar
  23. Kim SK, Son TK, Park SY, Lee IJ, Lee BH, Kim HY, Lee SC (2006) Influences of gibberellin and auxin on endogenous plant hormone and starch mobilization during rice seed germination under salt stress. J Environ Biol 27:181–186Google Scholar
  24. Lichfield CD (2002) Halotolerant bacteria. J Ind Microbiol Biotechnol 28:21–22Google Scholar
  25. Lin CC, Kao CH (2001) Cell wall peroxidise activity, hydrogen peroxide level and Nacl-inhibited root growth of rice seedlings. Plant Soil 230:135–143CrossRefGoogle Scholar
  26. Lucy M, Reed E, Glick BR (2004) Applications of free living plant growth-promoting rhizobacteria. Antonie van Leeuwenhoek 86:1–25PubMedCrossRefGoogle Scholar
  27. Madhaiyan M, Kim BY, Poonguzhali S, Kwon SW, Song MH, Ryu JH, Go SJ, Koo BS, Sa TM (2007) Methylobacterium oryzae sp. nov., a novel aerobic, pink-pigmented, facultatively methylotrophic, 1-aminocyclopropane-1-carboxylate deaminase producing bacterium isolated from rice. Int J Syst Evol Microbiol 57:326–331PubMedCrossRefGoogle Scholar
  28. Mayak S, Tirosh T, Glick BR (2004) Plant growth-promoting bacteria confer resistance in tomato plants to salt stress. Plant Physiol Biochem 42:565–572PubMedCrossRefGoogle Scholar
  29. Panda D, Sharma SG, Sarkar RK (2008) Chlorophyll fluorescence parameters, CO2 Photosynthetic rate and regeneration capacity as a result of complete submergence and subsequent re-emergence in rice (Oryza sativa L.). Aquat Bot 88:127–133CrossRefGoogle Scholar
  30. Penrose DM, Glick BR (2001) Levels of 1-aminocyclopropane-1-carboxylic acid (ACC) in exudates and extracts of canola seeds treated with plant growth-promoting bacteria. Can J Microbiol 47:368–372PubMedCrossRefGoogle Scholar
  31. Penrose DM, Glick BR (2003) Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria. Physiol Plant 118:10–15PubMedCrossRefGoogle Scholar
  32. Porra RJ (2002) The chequered history of the development and use of simultaneous equations for accurate determination of chlorophylls a and b. Photosynth Res 73:149–156PubMedCrossRefGoogle Scholar
  33. Raaijmakers JM, Paulitz TC, Steinberg C, Alabouvette C, Moënne-Loccoz T (2009) The rhizosphere: a playground and battlefield for soil-borne pathogens and beneficial microorganisms. Plant Soil 321:341–361CrossRefGoogle Scholar
  34. Ramakrishna C, Sethunathan N (1982) Stimulation of autotrophic ammonium oxidation in rice rhizosphere soil by the insecticide carbofuran. Appl Environ Microbiol 44:1–4PubMedGoogle Scholar
  35. Saleem M, Arshad M, Hussain S, Bhatti AS (2007) Perspective of plant growth promoting rhizobacteria (PGPR) containing ACC deaminase in stress agriculture. J Ind Microbiol Biotechnol 34:635–648PubMedCrossRefGoogle Scholar
  36. Sapsirisopa S, Chookietwattana K, Maneewan K and Khaengkhan P (2009) Effect of salt-tolerant Bacillus inoculum on rice KDML 105 cultivated in saline soil. Asian J Food Ag-Ind, S69–S74Google Scholar
  37. Saravanakumar D, Samiyappan R (2007) ACC deaminase from Pseudomonas fluorescens mediated saline resistance in groundnut (Arachis hypogea) plants. J Appl Microbiol 102:1283–1292PubMedCrossRefGoogle Scholar
  38. Sasser M (2001) Identification of Bacteria by Gas Chromatography of Cellular Fatty Acids. MIDI Technical Note 101Google Scholar
  39. Siddikee MA, Glick BR, Chauhan PS, Yim W, Sa T (2011) Enhancement of growth and salt tolerance of red pepper seedlings (Capsicum annuum L.) by regulating stress ethylene synthesis with halotolerant bacteria containing 1-aminocyclopropane-1-carboxylic acid deaminase activity. Plant Physiol Biochem 49:427–434PubMedCrossRefGoogle Scholar
  40. Spark DL, Page AL, Summer ME, Tabatabai MA, Helmke PA (1996) Methods of soil analysis, part 3, chemical methods. American Society of Agronomy, USAGoogle Scholar
  41. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  42. Yoshida S, Forno DA, Cock JH, Gomez KA (1976) Laboratory manual for physiological studies of rice, 3rd edn. International Rice Research Institutes, ManilaGoogle Scholar
  43. Zahir ZA, Muhammad A, Frankenberger WT Jr (2003) Plant growth promoting rhizobacteria: applications and perspectives in agriculture. Adv Agron 81:97–168CrossRefGoogle Scholar
  44. Zahir ZA, Ghani U, Naveed M, Nadeem SM, Asghar HN (2009) Comparative effectiveness of Pseudomonas and Serratia sp. containing ACC-deaminase for improving growth and yield of wheat (Triticum aestivum L.) under salt-stressed conditions. Arch Microbiol 191:415–424PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Himadri Bhusan Bal
    • 1
  • Lipika Nayak
    • 1
  • Subhasis Das
    • 1
  • Tapan K. Adhya
    • 1
    • 2
  1. 1.Laboratory of Soil Microbiology, Division of Crop ProductionCentral Rice Research InstituteCuttackIndia
  2. 2.School of BiotechnologyKIIT UniversityBhubaneswarIndia

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