Environmental Science and Pollution Research

, Volume 25, Issue 36, pp 36530–36544 | Cite as

Reduction in arsenic toxicity and uptake in rice (Oryza sativa L.) by As-resistant purple nonsulfur bacteria

  • Phitthaya Nookongbut
  • Duangporn KantachoteEmail author
  • Mallavarapu Megharaj
  • Ravi Naidu
Research Article


This study aimed to investigate the potential of Rhodopseudomonas palustris C1 and Rubrivivax benzoatilyticus C31 to ameliorate As toxicity and to reduce As uptake in rice. Strain C1 was superior to strain C31 for siderophore production. The mixed culture (1: 1) was most effective in reducing the toxicity of As species [As(III) and/or As(V), each 30 mg/l] by yielding maximal germination index that related to α- and β-amylase activities in two Thai rice cultivars (HomNil: HN and PathumThani 1: PT). Arsenic toxicity to the seed germination followed the order: mixed As species > As(III) > As(V); and the toxicity was reduced in inoculated sets, particularly with a mixed culture. The mixed culture significantly enhanced rice growth under As stress in both rice cultivars as indicated by an increase in the production of chlorophyll a and b, and also supporting the non-enzymatic (carotenoids, lipid oxidation, and nitric oxide) and enzymatic (superoxide dismutase, ascorbate peroxidase, catalase, and glutathione reductase) activities. These were concomitant with productions of 5-aminolevulinic acid, indole-3-acetic acid, exopolymeric substances, and siderophores which significantly reduced As accumulation in treated rice. It can be concluded that the mixed culture has great potential to ameliorate rice from As toxicity by preventing As species entry into rice for enhancing rice growth and also for reducing As accumulation to produce safe rice from rice grown in contaminated paddy fields.


Arsenic Contamination Plant growth-promoting substances Phototrophic bacteria Protection mechanisms Rice 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11356_2018_3568_MOESM1_ESM.docx (2.3 mb)
ESM 1 (DOCX 2400 kb)


  1. Ahmad P, Jaleel CA, Salem MA, Nabi G, Sharma S (2010) Roles of enzymatic and nonenzymatic antioxidants in plants during abiotic stress. Crit Rev Biotechnol 30(30):161–175CrossRefGoogle Scholar
  2. Ali B, Wang B, Ali S, Ghani MA, Hayat MT, Yang C, Xu L, Zhou WJ (2013) 5-Aminolevulinic acid ameliorates the growth, photosynthetic gas exchange capacity and ultrastructural changes under cadmium stress in Brassica napus L. J Plant Growth Regul 32:604–614CrossRefGoogle Scholar
  3. Armendariz AL, Talano MA, Wevar Oller AL, Medina MI, Agostini E (2015) Effect of arsenic on tolerance mechanisms of two plant growth-promoting bacteria used as biological inoculants. J Environ Sci (China) 33:203–210CrossRefGoogle Scholar
  4. Asada K (1984) Chloroplasts: formation of active oxygen and its scavenging. Methods Enzymol 105:422–429CrossRefGoogle Scholar
  5. Ashraf M, Harris PJC (2013) Photosynthesis under stressful environments: an overview. Photosynthetica 51(2):163–190CrossRefGoogle Scholar
  6. ATSDR (2015) CERCLA Priority List of Hazardous Substances for 2015. Accessed 29 August 2016
  7. Blodgett R (2010) BAM Appendix 2: Most Probable Number from Serial Dilutions. Accessed 22 January 2015
  8. Burnham BF (1970) δ-Aminolevulinic acid synthase (from Rhodopseudomonas sphaeroides). Methods Enzymol 17A:195–204CrossRefGoogle Scholar
  9. Chun-xi L, Shu-li F, Yun S, Li-na J, Xu-yang L, Xiao-li H (2007) Effects of arsenic on seed germination and physiological activities of wheat seedlings. J Environ Sci 19(6):725–732CrossRefGoogle Scholar
  10. Costache MA, Campeanu G, Neata G (2012) Studies concerning the extraction of chlorophyll and totalcarotenoids from vegetables. Rom Biotechnol Lett 17(5):7702–7708Google Scholar
  11. Dai YF, Xiao Y, Zhang EH, Liu LD, Qiu L, You LX, Dummi Mahadevan G, Chen BL, Zhao F (2016) Effective methods for extracting extracellular polymeric substances from Shewanella oneidensis MR-1. Water Sci Technol 74(12):2987–2996CrossRefGoogle Scholar
  12. Dhindsa RS, Plumb-Dhindsa P, Throne TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation and decreased levels of superoxide dismutase and catalase. J Exp Bot 32:93–101CrossRefGoogle Scholar
  13. Dimkpa CO, Merten D, Svatos A, Büchel G, Kothe E (2009) Metal-induced oxidative stress impacting plant growth in contaminated soil is alleviated by microbial siderophores. Soil Biol Biochem 41:154–162CrossRefGoogle Scholar
  14. Finnegan PM, Chen W (2012) Arsenic toxicity: the effects on plant metabolism. Front Physiol 3(182):1–18Google Scholar
  15. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48(12):909–930CrossRefGoogle Scholar
  16. Glick BR (2012) Plant growth-promoting Bacteria: mechanisms and applications. Scientifica 2012:1–15 Article 963401CrossRefGoogle Scholar
  17. Gordon SA, Weber RP (1951) Colorimetric estimation of indole acetic acid. Plant Physiol 26:192–195CrossRefGoogle Scholar
  18. Groß F, Durner J, Gaupels F (2013) Nitric oxide, antioxidants and prooxidants in plant defense responses. Front Plant Sci 4:1–15 Article 419CrossRefGoogle Scholar
  19. Grotto D, Santa Maria L, Valetini J, Paniz C, Schmitt G, Garcia SC, Pomblum VJ, Rocha JB, Farina M (2009) Importance of the lipid peroxidation biomarkers and methodological aspects for malondialdehyde quantification. Quim Nova 32:169–174CrossRefGoogle Scholar
  20. Havir EA, McHale NA (1987) Biochemical and developmental characterization of multiple forms of catalase in tobacco leaves. Plant Physiol 84:450–455CrossRefGoogle Scholar
  21. Jiang Z, Ma B, Erinle KO, Cao B, Liu X, Ye S, Zhang Y (2016) Enzymatic antioxidant defense in resistant plant: Pennisetuma mericanum (L.) K. Schum during long-term atrazine exposure. Pest Biochem Physiol 133:59–66CrossRefGoogle Scholar
  22. Kantachote D, Nunkaew T, Kantha T, Chaiprapat S (2016) Biofertilizers from Rhodopseudomonas palustris strains to enhance rice yields and reduce methane emissions. Appl Soil Ecol 100:154–161CrossRefGoogle Scholar
  23. Kantha T, Kantachote D, Klongdee N (2015) Potential of biofertilizers from selected Rhodopseudomonas palustris strains to assist rice (Oryza sativa L. subsp. indica) growth under salt stress and to reduce greenhouse gas emissions. Ann Microbiol 65:2109–2118CrossRefGoogle Scholar
  24. Karadeniz A, Topcuoğlu ŞF, Ìnan S (2006) Auxin, gibberellin, cytokinin and abscisic acid production in some bacteria. World J Microbiol Biotechnol 22(10):1061–1064CrossRefGoogle Scholar
  25. Li N, Wang J, Song WY (2016) Arsenic uptake and translocation in plants. Plant Cell Physiol 57:4–13CrossRefGoogle Scholar
  26. Liermann LJ, Kalinowski BE, Brantley SL, Ferry JG (2000) Role of bacterial siderophores in dissolution of hornblende. Geochim Cosmochim Acta 64:587–602CrossRefGoogle Scholar
  27. Liu T, Zhu LS, Wang JH, Wang J, Xie H (2015) The genotoxic and cytotoxic effects of 1-butyl-3-methylimidazolium chloride in soil on Vicia faba seedlings. J Hazard Mater 285:27–36CrossRefGoogle Scholar
  28. Lukasz D, Liwia R, Aleksandra M, Aleksandra S (2014) Dissolution of arsenic minerals mediated by dissimilatory arsenate reducing bacteria: estimation of the physiological potential for arsenic mobilization. Biomed Res Int 2014:841–892CrossRefGoogle Scholar
  29. Mishra S, Alfeld M, Sobotka R, Andresen E, Falkenberg G, Küpper H (2016) Analysis of sublethal arsenic toxicity to Ceratophyllum demersum: subcellular distribution of arsenic and inhibition of chlorophyll biosynthesis. J Exp Bot 67(15):4639–4646CrossRefGoogle Scholar
  30. Morita Y, Aibara S, Yamashita H, Yagi F, Suganama T, Hiromi K (1975) Crystallisation and preliminary X-ray investigation of soy bean β-amylase. J Biochem 77:343–351CrossRefGoogle Scholar
  31. Nair A, Juwarkar AA, Singh SK (2007) Production and characterization of siderophores and its application in arsenic removal from contaminated soil. Water Air Soil Pollut 180:199–212CrossRefGoogle Scholar
  32. Noctor G, Gomez L, Vanacker H, Foyer CH (2002) Interactions between biosynthesis, compartmentation and transport in the control of glutathione homeostasis and signalling. J Exp Bot 53:1283–1304CrossRefGoogle Scholar
  33. Nookongbut P, Kantachote D, Megharaj M (2016) Arsenic contamination in areas surrounding mines and selection of potential as-resistant purple nonsulfur bacteria for use in bioremediation based on their detoxification mechanisms. Ann Microbiol 66(4):1419–1429CrossRefGoogle Scholar
  34. Nookongbut N, Kantachote D, Krishnan K, Megharaj M (2017) Arsenic resistance genes of as-resistant purple nonsulfur bacteria isolated from as-contaminated sites for bioremediation application. J Basic Microbiol 57(4):316–324CrossRefGoogle Scholar
  35. Nordstrom DK (2002) Worldwide occurrences of arsenic in the ground water. Science 296(5567):2143–2145CrossRefGoogle Scholar
  36. Nunkaew T, Kantachote D, Kanzaki H, Nitoda T, Ritchie RJ (2014) Effects of 5-aminolevulinic acid (ALA)- containing supernatants from selected Rhodopseudomonas palustris strains on rice growth under NaCl stress, with mediating effects on chlorophyll, photosynthetic electron transport and antioxidative enzymes. Electron J Biotechnol 17(1):19–26CrossRefGoogle Scholar
  37. Nunkaew T, Kantachote D, Nitoda T, Kanzaki H, Ritchie RJ (2015) Characterization of exopolymeric substances from selected Rhodopseudomonas palustris strains and their ability to adsorb sodium ions. Carbohydr Polym 115:334–341CrossRefGoogle Scholar
  38. Paula JFR, Froes-Silva RES, Ciminelli VST (2012) Arsenic determination in complex mining residues by ICP OES after ultrasonic extraction. Microchem J 104:12–16CrossRefGoogle Scholar
  39. Raab A, Williams PN, Megharg A, Feldmann J (2007) Uptake and translocation of inorganic and methylated arsenic species by plants. Environ Chem 4:197–203CrossRefGoogle Scholar
  40. Raj A, Pandey AK, Sharma YK, Khare PB, Srivastava PK, Singh N (2011) Metabolic adaptation of Pteris vittata L. gametophyte to arsenic induced oxidative stress. Bioresour Technol 102(20):9827–9832CrossRefGoogle Scholar
  41. Rajkumar M, Ae N, Prasad MN, Freitas H (2010) Potential of siderophore-producing bacteria for improving heavy metal phytoextraction. Trends Biotechnol 28(3):142–149CrossRefGoogle Scholar
  42. Sakpirom J, Kantachote D, Nunkaew T, Khan E (2017) Characterizations of purple nonsulfur bacteria isolated from paddy fields and identification of strains with potential for plant growth promotion, greenhouse gas mitigation and heavy metal bioremediation. Res Microbiol 168(3):266–275CrossRefGoogle Scholar
  43. Sandalio LM, Rodríguez-Serrano M, del Río LA, Romero-puertas MC (2009) Reactive oxygen species and signalling in cadmium toxicity. In: del Río LA, Puppo A (eds) Reactive oxygen species in plant Signalling. Springer, Heidelberg, pp 175–190CrossRefGoogle Scholar
  44. Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56CrossRefGoogle Scholar
  45. Shaik SS, Carciofi M, Martens HJ, Hebelstrup KH, Blennow A (2014) Starch bioengineering affects cereal grain germination and seedling establishment. J Exp Bot 65(9):2257–2270CrossRefGoogle Scholar
  46. Shain Y, Mayer AM (1968) Activation of enzyme during germination: amylopectin 1, 6-glucosidase in pea. Physiol Plant 21:765–771CrossRefGoogle Scholar
  47. Singh HP, Batish DR, Kohli RK, Arora K (2007) Arsenic-induced root growth inhibition in mung bean (Phaseolus aureus Roxb.) is due to oxidative stress resulting from enhanced lipid peroxidation. Plant Growth Regul 53(1):65–73CrossRefGoogle Scholar
  48. Song NH, Yin XL, Chen GF, Yang H (2007) Biological responses of wheat (Triticuma estivum) plants to the herbicide chlorotoluron in soils. Chemosphere 68:1779–1787CrossRefGoogle Scholar
  49. Sun J, Zhang X, Broderick M, Fein H (2003) Measurement of nitric oxide production in biological systems by using Griess reaction assay. Sensors 3:276–284CrossRefGoogle Scholar
  50. Sun Z, Liu Y, Huang Y, Zeng G, Wang Y, Hu X, Zhou L (2014) Effects of indole-3-acetic, kinetin and spermidine assisted with EDDS on metal accumulation and tolerance mechanisms in ramie (Boehmeria nivea (L.) gaud.). Ecol Eng 71:108–112CrossRefGoogle Scholar
  51. Tripathi M, Munot HP, Shouche Y, Meyer JM, Goel R (2005) Isolation and functional characterization of siderophore-producing lead- and cadmium-resistant Pseudomonas putida KNP9. Curr Microbiol 50(5):233–237CrossRefGoogle Scholar
  52. Tripathi RD, Srivastava S, Mishra S, Singh N, Tuli R, Gupta DK, Maathuis FJ (2007) Arsenic hazards: strategies for tolerance and remediation by plants. Trends Biotechnol 25(4):158–165CrossRefGoogle Scholar
  53. Tripathi RD, Tripathi P, Dwivedi S, Dubey S, Chatterjee S, Chakrabarty D, Trivedi PK (2012) Arsenomics: omics of arsenic metabolism in plants. Front Plant Physiol 3:1–14Google Scholar
  54. Tripathi P, Singh RP, Sharma YK, Tripathi RD (2015) Arsenite stress variably stimulates pro-oxidant enzymes, anatomical deformities, photosynthetic pigment reduction, and antioxidants in arsenic-tolerant and sensitive rice seedlings. Environ Toxicol Chem 34(7):1562–1571CrossRefGoogle Scholar
  55. Ünyayar S, Topcuoğlu ŞF, Ünyayar A (1996) A modified method for extraction and identification of indole-3-acetic acid (IAA),gibberellic acid (GA3), abscisic acid (ABA) and zeatin produced by Phanerochaete chrysosporium, ME 446. Bulg J Plant Physiol 22(3–4):105–110Google Scholar
  56. Upadhyay AK, Singh NK, Rai UN (2014) Comparative metal accumulation potential of Potamogeton Pectinatus L. and Potamogeton crispus L.: role of enzymatic and non-enzymatic antioxidants in tolerance and detoxification of metals. Aquat Bot 117:27–32CrossRefGoogle Scholar
  57. Upadhyay AK, Singh NK, Singh R, Rai UN (2016) Amelioration of arsenic toxicity in rice: comparative effect of inoculation of Chlorella vulgaris and Nannochloropsis sp. on growth, biochemical changes and arsenic uptake. Ecotoxicol Environ Saf 124:68–73CrossRefGoogle Scholar
  58. Van Breusegem F, Vranová E, Dat JF, Inzé D (2001) The role of active oxygen species in plant signal transduction. Plant Sci 161(3):405–414CrossRefGoogle Scholar
  59. Wang M, Zhou Q (2006) Effects of herbicide chlorimuron-ethyl on physiological mechanisms in wheat (Triticuma estivum). Ecotoxicol Environ Saf 64(2):190–197CrossRefGoogle Scholar
  60. Wellburn AR (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313CrossRefGoogle Scholar
  61. Yang GD, Xie WY, Zhu X, Huang Y, Yang XJ, Qiu ZQ, Lv ZM, Wang WN, Lin WX (2015) Effect of arsenite-oxidizing bacterium B. laterosporus on arsenite toxicity and arsenic translocation in rice seedlings. Ecotoxicol Environ Saf 120:7–12CrossRefGoogle Scholar
  62. Yu HY, Liu C, Zhu J, Li F, Deng DM, Wang Q, Liu C (2016) Cadmium availability in rice paddy fields from a mining area: the effects of soil properties highlighting iron fractions and pH value. Environ Pollut 209:38–45CrossRefGoogle Scholar
  63. Zhang C, Ge Y, Yao H, Chen X, Hu M (2012) Iron oxidation-reduction and its impacts on cadmium bioavailability in paddy soils: a review. Front Environ Sci Eng 6:509–517CrossRefGoogle Scholar
  64. Zhao FJ, McGrath SP, Meharg AA (2010) Arsenic as a food chain contaminant: mechanisms of plant uptake and metabolism and mitigation strategies. Annu Rev Plant Biol 61:535–559CrossRefGoogle Scholar
  65. Zhu YG, William PN, Megharg AA (2008) Exposure to inorganic arsenic from rice: a global health issue? Environ Pollut 154:169–171CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Phitthaya Nookongbut
    • 1
  • Duangporn Kantachote
    • 1
    • 2
    Email author
  • Mallavarapu Megharaj
    • 3
    • 4
  • Ravi Naidu
    • 3
    • 4
  1. 1.Department of Microbiology, Faculty of SciencePrince of Songkla UniversityHat YaiThailand
  2. 2.Center of Excellence on Hazardous Substance Management (HSM)BangkokThailand
  3. 3.Global Centre for Environmental Remediation, Faculty of ScienceThe University of NewcastleCallaghanAustralia
  4. 4.Cooperative Research Centre for Contamination Assessment and Remediation of the Environment (CRC-CARE)The University of NewcastleCallaghanAustralia

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