Plant and Soil

, Volume 362, Issue 1–2, pp 301–318 | Cite as

Hydrogen sulfide alleviates aluminum toxicity in barley seedlings

  • Juan Chen
  • Wen-Hua Wang
  • Fei-Hua Wu
  • Chun-Yan You
  • Ting-Wu Liu
  • Xue-Jun Dong
  • Jun-Xian He
  • Hai-Lei ZhengEmail author
Regular Article



Aluminum (Al) toxicity is one of the major factors that limit plant growth. Low concentration of hydrogen sulfide (H2S) has been proven to function in physiological responses to various stresses. The objective of this study is to investigate the possible role of H2S in Al toxicity in barley (Hordeum vulgare L) seedlings.


Barley seedlings pre-treated with sodium hydrosulfide (NaHS), a H2S donor, and subsequently exposed to Al treatment were studied for their effects on root elongation, Al accumulation in seedlings, Al-induced citrate secretion and oxidative stress, and plasma membrane (PM) H+-ATPase expression.


Our results showed that H2S had significant rescue effects on Al-induced inhibition of root elongation which was correlated well with the decrease of Al accumulation in seedlings. Meanwhile, Al-induced citrate secretion was also significantly enhanced by NaHS pretreatment. Al-induced oxidative stress as indicated by lipid peroxidation and reactive oxygen species burst was alleviated by H2S through the activation of the antioxidant system. Moreover, Al-induced reduction in PM H+-ATPase expression was reversed by exogenous NaHS.


Altogether, our results suggest H2S plays an ameliorative role in protecting plants against Al toxicity by inducing the activities of antioxidant enzymes, increasing citrate secretion and citrate transporter gene expression, and enhancing the expression of PM H+-ATPase.


Aluminum toxicity Citrate secretion H+-ATPase Hydrogen sulfide Oxidative stress Root elongation 



We are grateful to Bing-Bo Li, Duan-Ye Xiong, Xiang Liu and Jing Huang for assistance in experiments, and Mr. Sieh Sorie Kargbo for critically reading the manuscript. This study was financially supported by the Natural Science Foundation of China (NSFC) (30930076, 30770192, 30670317), the Foundation of the Chinese Ministry of Education (20070384033), the Program for New Century Excellent Talents in Xiamen University (NCETXMU X071l5) and a Changjiang Scholarship (X09111), the Scholarship award for excellent doctoral student granted by Ministry of Education.

Supplementary material

11104_2012_1275_MOESM1_ESM.doc (148 kb)
Esm 1 (DOC 147 kb)


  1. Ahn SJ, Rengel Z, Matsumoto H (2004) Aluminum-induced plasma membrane surface potential and H+-ATPase activity in near-isogenic wheat lines differing in tolerance to aluminum. New Phytologist 162:71–79CrossRefGoogle Scholar
  2. Ahn SJ, Sivaguru M, Chung GC, Rengel Z, Matsumoto H (2002) Aluminium-induced growth inhibition is associated with impaired efflux and influx of H+ across the plasma membrane in root apices of squash (Cucurbita pepo). Journal of Experimental Botany 53:1959–1966PubMedCrossRefGoogle Scholar
  3. Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant, Cell & Environment 24:1337–1344CrossRefGoogle Scholar
  4. Barcelo J, Poschenrieder C (2002) Fast root growth responses, root exudates, and internal detoxification as clues to the mechanisms of aluminium toxicity and resistance: a review. Environmental and Experimental Botany 48:75–92CrossRefGoogle Scholar
  5. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry 44:276–287PubMedCrossRefGoogle Scholar
  6. Behl C, Davis JB, Lesley R, Schubert D (1994) Hydrogen peroxide mediates amyloid β protein toxicity. Cell 77:817–827PubMedCrossRefGoogle Scholar
  7. Bennet R, Breen C (1991) The aluminium signal: new dimensions to mechanisms of aluminium tolerance. Plant and Soil 134:153–166Google Scholar
  8. Boscolo PRS, Menossi M, Jorge RA (2003) Aluminum-induced oxidative stress in maize. Phytochem 62:181–189CrossRefGoogle Scholar
  9. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254PubMedCrossRefGoogle Scholar
  10. Cao ZY, Xuan W, Liu ZY, Li XN, Zhao N, Xu P, Wang Z, Guan RZ, Shen WB (2007) Carbon monoxide promotes lateral root formation in rapeseed. Journal of Integrative Plant Biology 49:1070–1079CrossRefGoogle Scholar
  11. Cartes P, Jara AA, Pinilla L, Rosas A, Mora ML (2010) Selenium improves the antioxidant ability against aluminium-induced oxidative stress in ryegrass roots. Annals of Applied Biology 156:297–307CrossRefGoogle Scholar
  12. Chen J, Wu F-H, Wang W-H, Zheng C-J, Lin G-H, Dong X-J, He J-X, Pei Z-M, Zheng H-L (2011) Hydrogen sulphide enhances photosynthesis through promoting chloroplast biogenesis, photosynthetic enzyme expression, and thiol redox modification in Spinacia oleracea seedlings. Journal of Experimental Botany 62:4481–4493PubMedCrossRefGoogle Scholar
  13. Chen L, Wu F-H, Liu T-W, Chen J, Li Z-J, Pei Z-M, Zheng H-L (2010) Soil acidity reconstruction based on tree ring information of a dominant species Abies fabri in the subalpine forest ecosystems in southwest China. Environmental Pollution 158:3219–3224PubMedCrossRefGoogle Scholar
  14. Corrales I, Poschenrieder C, Barcelo J (2008) Boron-induced amelioration of aluminium toxicity in a monocot and a dicot species. Journal of Plant Physiology 165:504–513PubMedCrossRefGoogle Scholar
  15. Delhaize E, Ryan PR (1995) Aluminum toxicity and tolerance in plants. Plant Physiology 107:315–321PubMedGoogle Scholar
  16. Devi SR, Prasad MNV (1998) Copper toxicity in Ceratophyllum demersum L. (Coontail), a free floating macrophyte: response of antioxidant enzymes and antioxidants. Plant Science 138:157–165CrossRefGoogle Scholar
  17. Dhindsa RS, Plumb-Dhindsa P, Thorpe TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. Journal of Experimental Botany 32:93–101CrossRefGoogle Scholar
  18. Dunand C, Crevecoeur M, Penel C (2007) Distribution of superoxide and hydrogen peroxide in Arabidopsis root and their influence on root development: possible interaction with peroxidases. New Phytologist 174:332–341PubMedCrossRefGoogle Scholar
  19. Ezaki B, Gardner RC, Ezaki Y, Matsumoto H (2000) Expression of aluminum-induced genes in transgenic Arabidopsis plants can ameliorate aluminum stress and/or oxidative stress. Plant Physiology 122:657–665PubMedCrossRefGoogle Scholar
  20. Furukawa J, Yamaji N, Wang H, Mitani N, Murata Y, Sato K, Katsuhara M, Takeda K, Ma JF (2007) An aluminum-activated citrate transporter in barley. Plant & Cell Physiology 48:1081–1091CrossRefGoogle Scholar
  21. García-Mata C, Lamattina L (2010) Hydrogen sulphide, a novel gasotransmitter involved in guard cell signalling. New Phytologist 188:977–984PubMedCrossRefGoogle Scholar
  22. Hamilton CA, Good AG, Taylor GJ (2001) Induction of vacuolar ATPase and mitochondrial ATP synthase by aluminum in an aluminum-resistant cultivar of wheat. Plant Physiology 125:2068–2077PubMedCrossRefGoogle Scholar
  23. Hosoki R, Matsuki N, Kimura H (1997) The possible role of hydrogen sulfide as an endogenous smooth muscle relaxant in synergy with nitric oxide. Biochemical and Biophysical Research Communications 237:527–531PubMedCrossRefGoogle Scholar
  24. Hossain MA, Hossain AKMZ, Kihara T, Koyama H, Hara T (2005) Aluminum-induced lipid peroxidation and lignin deposition are associated with an increase in H2O2 generation in wheat seedlings. Soil Science & Plant Nutrition 51:223–230CrossRefGoogle Scholar
  25. Hsu YT, Kao CH (2004) Cadmium toxicity is reduced by nitric oxide in rice leaves. Plant Growth Regulation 42:227–238CrossRefGoogle Scholar
  26. Kinraide T (1991) Identity of the rhizotoxic aluminium species. Plant and Soil 134:167–178Google Scholar
  27. Kobayashi, Y., Yamamoto, Y., & Matsumoto, H. (2003). Aluminum-triggered production of superoxide anions and its possible involvement in root elongation inhibition in pea (Pisum sativum). Plant Cell Physiol, 44(Supplement), 165–165.Google Scholar
  28. Kochian LV (1995) Cellular mechanisms of aluminum toxicity and resistance in plants. Annual Review of Plant Physiology and Plant Molecular Biology 46:237–260CrossRefGoogle Scholar
  29. Kochian LV, Hoekenga OA, Pineros MA (2004) How do crop plants tolerate acid soils?—Mechanisms of aluminum tolerance and phosphorous efficiency. Annual Review of Plant Biology 55:459–493PubMedCrossRefGoogle Scholar
  30. Kochian LV, Piñeros MA, Hoekenga OA (2005) The physiology, genetics and molecular biology of plant aluminum resistance and toxicity. Plant and Soil 274:175–195CrossRefGoogle Scholar
  31. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  32. Larson RA (1988) The antioxidants of higher plants. Phytochem 27:969–978CrossRefGoogle Scholar
  33. Li D, Xiao Z, Liu L, Wang J, Song G, Bi Y (2010) Effects of exogenous hydrogen sulfide (H2S) on the root tip an root border cells of Pisum sativum. Chin Bull Bot 45:354–362 (in Chinese with English abstract)Google Scholar
  34. Li L, Bhatia M, Moore PK (2006) Hydrogen sulphide—a novel mediator of inflammation? Current Opinion in Pharmacology 6:125–129PubMedCrossRefGoogle Scholar
  35. Ligaba A, Yamaguchi M, Shen H, Sasaki T, Yamamoto Y, Matsumoto H (2004) Phosphorus deficiency enhances plasma membrane H+-ATPase activity and citrate exudation in greater purple lupin (Lupinus pilosus). Functional Plant Biology 31:1075–1083CrossRefGoogle Scholar
  36. Lisjak M, Srivastava N, Teklic T, Civale L, Lewandowski K, Wilson I, Wood ME, Whiteman M, Hancock JT (2010) A novel hydrogen sulfide donor causes stomatal opening and reduces nitric oxide accumulation. Plant Physiology and Biochemistry 48:931–935PubMedCrossRefGoogle Scholar
  37. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt. Methods 25:402–408PubMedCrossRefGoogle Scholar
  38. Magalhaes JV, Liu J, Guimaraes CT, Lana UGP, Alves VMC, Wang Y-H, Schaffert RE, Hoekenga OA, Pineros MA, Shaff JE, Klein PE, Carneiro NP, Coelho CM, Trick HN, Kochian LV (2007) A gene in the multidrug and toxic compound extrusion (MATE) family confers aluminum tolerance in sorghum. Nature Genetics 39:1156–1161PubMedCrossRefGoogle Scholar
  39. Ma JF (2000) Role of organic acids in detoxification of aluminum in higher plants. Plant & Cell Physiology 41:389–390Google Scholar
  40. Mata CG, Lamattina L (2001) Nitric oxide induces stomatal closure and enhances the adaptive plant responses against drought stress. Plant Physiology 126:1196–1204CrossRefGoogle Scholar
  41. Matsumoto H (2000) Cell biology of aluminum toxicity and tolerance in higher plants. International Review of Cytology 200:1–46PubMedCrossRefGoogle Scholar
  42. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science 7:405–410PubMedCrossRefGoogle Scholar
  43. Morsomme P, Boutry M (2000) The plant plasma membrane H+-ATPase: structure, function and regulation. Biochimica et Biophysica Acta 1465:1–16PubMedCrossRefGoogle Scholar
  44. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant & Cell Physiology 22:867–880Google Scholar
  45. Ohno T, Koyama H, Hara T (2003) Characterization of citrate transport through the plasma membrane in a carrot mutant cell line with enhanced citrate excretion. Plant & Cell Physiology 44:156–162CrossRefGoogle Scholar
  46. Ohno T, Nakahira S, Suzuki Y, Kani T, Hara T, Koyama H (2004) Molecular characterization of plasma membrane H+-ATPase in a carrot mutant cell line with enhanced citrate excretion. Physiologia Plantarum 122:265–274CrossRefGoogle Scholar
  47. Ryan PR, Ditomaso JM, Kochian LV (1993) Aluminium toxicity in roots: an investigation of spatial sensitivity and the role of the root cap. Journal of Experimental Botany 44:437–446CrossRefGoogle Scholar
  48. Ryan P, Delhaize E, Randall P (1995a) Malate efflux from root apices and tolerance to aluminium are highly correlated in wheat. Functional Plant Biology 22:531–536Google Scholar
  49. Ryan PR, Delhaize E, Randall PJ (1995b) Characterisation of Al-stimulated efflux of malate from the apices of Al-tolerant wheat roots. Planta 196:103–110CrossRefGoogle Scholar
  50. Ryan PR, Tyerman SD, Sasaki T, Furuichi T, Yamamoto Y, Zhang WH, Delhaize E (2010) The identification of aluminium-resistance genes provides opportunities for enhancing crop production on acid soils. Journal of Experimental Botany 62:9–20PubMedCrossRefGoogle Scholar
  51. Sasaki M, Yamamoto Y, Ma JF, Matsumoto H (1997) Early events induced by aluminum stress in elongating cells of wheat root. Soil Science & Plant Nutrition 43:1009–1014CrossRefGoogle Scholar
  52. Shen H, He LF, Sasaki T, Yamamoto Y, Zheng SJ, Ligaba A, Yan XL, Ahn SJ, Yamaguchi M, Sasakawa H, Matsumoto H (2005) Citrate secretion coupled with the modulation of soybean root tip under aluminum stress. Up-regulation of transcription, translation, and threonine-oriented phosphorylation of plasma membrane H+-ATPase. Plant Physiology 138:287–296PubMedCrossRefGoogle Scholar
  53. Sussman MR (1994) Molecular analysis of proteins in the plant plasma membrane. Annual Review of Plant Physiology and Plant Molecular Biology 45:211–234CrossRefGoogle Scholar
  54. Tahara K, Yamanoshita T, Norisada M, Hasegawa I, Kashima H, Sasaki S, Kojima K (2008) Aluminum distribution and reactive oxygen species accumulation in root tips of two Melaleuca trees differing in aluminum resistance. Plant and Soil 307:167–178CrossRefGoogle Scholar
  55. Tamas L, Simonovicova M, Huttova J, Mistrik I (2004) Aluminium stimulated hydrogen peroxide production of germinating barley seeds. Environmental and Experimental Botany 51:281–288CrossRefGoogle Scholar
  56. Tian Q-Y, Sun D-H, Zhao M-G, Zhang W-H (2007) Inhibition of nitric oxide synthase (NOS) underlies aluminum-induced inhibition of root elongation in Hibiscus moscheutos. New Phytologist 174:322–331PubMedCrossRefGoogle Scholar
  57. Uexküll HR, Mutert E (1995) Global extent, development and economic impact of acid soils. Plant and Soil 171:1–15CrossRefGoogle Scholar
  58. Wang B-L, Shi L, Li Y-X, Zhang W-H (2010) Boron toxicity is alleviated by hydrogen sulfide in cucumber (Cucumis sativus L.) seedlings. Planta 231:1301–1309PubMedCrossRefGoogle Scholar
  59. Wang R (2002) Two’s company, three’s a crowd: can H2S be the third endogenous gaseous transmitter? The FASEB Journal 16:1792–1798CrossRefGoogle Scholar
  60. Wang Y-S, Yang Z-M (2005) Nitric oxide reduces aluminum toxicity by preventing oxidative stress in the roots of Cassia tora L. Plant & Cell Physiology 46:1915–1923CrossRefGoogle Scholar
  61. Xuan W, Zhu FY, Xu S, Huang BK, Ling TF, Qi JY, Ye MB, Shen WB (2008) The heme oxygenase/carbon monoxide system is involved in the auxin-induced cucumber adventitious rooting process. Plant Physiology 148:881–893PubMedCrossRefGoogle Scholar
  62. Yamamoto Y, Kobayashi Y, Devi SR, Rikiishi S, Matsumoto H (2002) Aluminum toxicity is associated with mitochondrial dysfunction and the production of reactive oxygen species in plant cells. Plant Physiology 128:63–72PubMedCrossRefGoogle Scholar
  63. Yamamoto Y, Kobayashi Y, Devi SR, Rikiishi S, Matsumoto H (2003) Oxidative stress triggered by aluminum in plant roots. Plant and Soil 255:239–243CrossRefGoogle Scholar
  64. Yan F, Feuerle R, Schaffer S, Fortmeier H, Schubert S (1998) Adaptation of active proton pumping and plasmalemma ATPase activity of corn roots to low root medium pH. Plant Physiology 117:311–319PubMedCrossRefGoogle Scholar
  65. Yan K, Chen W, Zhang GY, Xu S, Liu ZL, He XY, Wang LL (2010) Elevated CO2 ameliorated oxidative stress induced by elevated O3 in Quercus mongolica. Acta Physiol Plant 32:375–385CrossRefGoogle Scholar
  66. Yang G, Wu L, Jiang B, Yang W, Qi J, Cao K, Meng Q, Mustafa AK, Mu W, Zhang S, Snyder SH, Wang R (2008) H2S as a physiologic vasorelaxant: hypertension in mice with deletion of cystathionine γ -lyase. Science 322:587–590PubMedCrossRefGoogle Scholar
  67. Yang JL, You JF, Li YY, Wu P, Zheng SJ (2007) Magnesium enhances aluminum-induced citrate secretion in rice bean roots (Vigna umbellata) by restoring plasma membrane H+-ATPase activity. Plant & Cell Physiology 48:66–73CrossRefGoogle Scholar
  68. Zhao ZQ, Ma JF, Sato K, Takeda K (2003) Differential Al resistance and citrate secretion in barley (Hordeum vulgare L.). Planta 217:794–800PubMedCrossRefGoogle Scholar
  69. Zhang H, Hu L-Y, Hu K-D, He Y-D, Wang S-H, Luo J-P (2008) Hydrogen sulfide promotes wheat seed germination and alleviates oxidative damage against copper stress. Journal of Integrative Plant Biology 50:1518–1529PubMedCrossRefGoogle Scholar
  70. Zhang H, Hu LY, Li P, Hu KD, Jiang CX, Luo JP (2010a) Hydrogen sulfide alleviated chromium toxicity in wheat. Biologia Plantarum 54:743–747CrossRefGoogle Scholar
  71. Zhang H, Jiao H, Jiang C-X, Wang S-H, Wei Z-J, Luo J-P, Jones R (2010b) Hydrogen sulfide protects soybean seedlings against drought-induced oxidative stress. Acta Physiol Plant 32:849–857CrossRefGoogle Scholar
  72. Zhang H, Tan Z-Q, Hu L-Y, Wang S-H, Luo J-P, Jones RL (2010c) Hydrogen sulfide alleviates aluminum toxicity in germinating wheat seedlings. Journal of Integrative Plant Biology 52:556–567PubMedCrossRefGoogle Scholar
  73. Zhang H, Ye Y-K, Wang S-H, Luo J-P, Tang J, Ma D-F (2009a) Hydrogen sulfide counteracts chlorophyll loss in sweetpotato seedling leaves and alleviates oxidative damage against osmotic stress. Plant Growth Regulation 58:243–250CrossRefGoogle Scholar
  74. Zhang S, Lu S, Xu X, Korpelainen H, Li C (2009b) Changes in antioxidant enzyme activities and isozyme profiles in leaves of male and female Populus cathayana infected with Melampsora larici-populina. Tree Physiology 30:116–128PubMedCrossRefGoogle Scholar
  75. Zheng SJ, Ma JF, Matsumoto H (1998) High aluminium resistance in buckwheat. I. Al-induced specific secretion of oxalic acid from root tips. Plant Physiology 117:745–751CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Juan Chen
    • 1
  • Wen-Hua Wang
    • 1
  • Fei-Hua Wu
    • 1
  • Chun-Yan You
    • 1
  • Ting-Wu Liu
    • 1
  • Xue-Jun Dong
    • 2
  • Jun-Xian He
    • 3
  • Hai-Lei Zheng
    • 1
    Email author
  1. 1.Key Laboratory for Subtropical Wetland Ecosystem Research of MOE, College of the Environment and EcologyXiamen UniversityXiamenPeople’s Republic of China
  2. 2.Central Grasslands Research Extension CenterNorth Dakota State UniversityStreeterUSA
  3. 3.State Key Laboratory of Agobiotechnology and School of Life SciencesThe Chinese University of Hong KongHong KongPeople’s Republic of China

Personalised recommendations