Skip to main content
SpringerLink
Log in

Hydrogen sulfide alleviates zinc toxicity by reducing zinc uptake and regulating genes expression of antioxidative enzymes and metallothioneins in roots of the cadmium/zinc hyperaccumulator Solanum nigrum L.

  • Regular Article
  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

Aims

Zinc (Zn) is a toxic element for plant at high concentrations. As a biologically active gaseous molecule, hydrogen sulfide (H2S) regulates plant growth and development. The aim of this study is to reveal the ameliorative effects of H2S on the physiological and molecular responses of a hyperaccumulator to Zn toxicity.

Methods

Growth analysis, biochemical and zymological methods, fluorescent microscopic imaging, western-blot and quantitative real-time PCR analysis were performed to investigate the roles of NaHS (a donor of H2S) on the cadmium (Cd)/Zn hyperaccumulator, Solanum nigrum L.

Results

H2S ameliorated excess Zn-induced growth inhibition, especially in roots. H2S decreased free cytosolic Zn2+ content in roots which was correlated well with the down-regulation of Zn uptake and homeostasis related genes expression. Besides, H2S further enhanced the expression of the metallothioneins (MTs). Moreover, Zn-induced oxidative stress was also alleviated by H2S.

Conclusions

The alleviation of H2S on excess Zn toxicity in S. nigrum is presumably attributed to: (1) the decrease in Zn accumulation via down-regulation of Zn uptake and homeostasis related genes expression; (2) the enhancement in the expression of MTs which chelate excess Zn; (3) the change in the genes expression of antioxidative enzymes, leading to H2O2 reduction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Adams JP, Adeli A, Hsu CY, Harkess RL, Page GP, dePamphilis CW, Schultz EB, Yuceer C (2011) Poplar maintains zinc homeostasis with heavy metal genes HMA4 and PCS1. J Exp Bot 62:3737–3752

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • 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 Environ 24:1337–1344

    Article  CAS  Google Scholar 

  • Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

    Article  CAS  PubMed  Google Scholar 

  • Arrivault S, Senger T, Krämer U (2006) The Arabidopsis metal tolerance protein AtMTP3 maintains metal homeostasis by mediating Zn exclusion from the shoot under Fe deficiency and Zn oversupply. Plant J 46:861–879

    Article  CAS  PubMed  Google Scholar 

  • Baker A, Reeves R, Hajar A (2006) Heavy metal accumulation and tolerance in British populations of the metallophyte Thlaspi caerulescens J. & C. Presl (Brassicaceae). New Phytol 127:61–68

    Article  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Broadley MR, White PJ, Hammond JP, Zelko I, Lux A (2007) Zinc in plants. New Phytol 173:677–702

    Article  CAS  PubMed  Google Scholar 

  • Cakmak I (2000) Possible roles of zinc in protecting plant cells from damage by reactive oxygen species. New Phytol 146:185–205

    Article  CAS  Google Scholar 

  • Chen J, Wu FH, Wang WH, Zheng CJ, Lin GH, Dong XJ, He JX, Pei ZM, Zheng HL (2011) Hydrogen sulphide enhances photosynthesis through promoting chloroplast biogenesis, photosynthetic enzyme expression, and thiol redox modification in Spinacia oleracea seedlings. J Exp Bot 62:4481–4493

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Chen J, Wang WH, Wu FH, You CY, Liu TW, Dong XJ, He JX, Zheng HL (2012) Hydrogen sulfide alleviates aluminum toxicity in barley seedlings. Plant Soil 362:301–318

    Article  Google Scholar 

  • Cherian M, Chan H (1993) Biological functions of metallothionein: a review. In: Suzuki KT, Imura N, Kimura M (eds) Metallothionein III: biological roles and medical implications. Birkhauser, Basel, pp. 87–109

    Google Scholar 

  • Clemens S (2001) Molecular mechanisms of plant metal tolerance and homeostasis. Planta 212:475–486

    Article  CAS  PubMed  Google Scholar 

  • Cobbett C, Goldsbrough P (2002) Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostasis. Annu Rev Plant Biol 53:159–182

    Article  CAS  PubMed  Google Scholar 

  • Courbot M, Willems G, Motte P, Arvidsson S, Roosens N, Saumitou-Laprade P, Verbruggen N (2007) A major quantitative trait locus for cadmium tolerance in Arabidopsis halleri colocalizes with HMA4, a gene encoding a heavy metal ATPase. Plant Physiol 144:1052–1065

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Drager DB, Desbrosses-Fonrouge AG, Krach C, Chardonnens AN, Meyer RC, Saumitou-Laprade P, Kramer U (2004) Two genes encoding Arabidopsis halleri MTP1 metal transport proteins co-segregate with zinc tolerance and account for high MTP1 transcript levels. Plant J 39:425–439

    Article  PubMed  Google Scholar 

  • Ferraz P, Fidalgo F, Almeida A, Teixeira J (2012) Phytostabilization of nickel by the zinc and cadmium hyperaccumulator Solanum nigrum L. Are metallothioneins involved? Plant Physiol Biochem 57:254–260

    Article  CAS  PubMed  Google Scholar 

  • Fidalgo F, Azenha M, Silva AF, de Sousa A, Santiago A, Ferraz P, Teixeira J (2013) Copper-induced stress in Solanum nigrum L. and antioxidant defense system responses. Food Energy Sec 2:70–80

    Article  Google Scholar 

  • Fukao Y, Ferjani A, Tomioka R, Nagasaki N, Kurata R, Nishimori Y, Fujiwara M, Maeshima M (2011) ITRAQ analysis reveals mechanisms of growth defects due to excess zinc in Arabidopsis. Plant Physiol 155:1893–1907

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Grotz N, Fox T, Connolly E, Park W, Guerinot ML, Eide D (1998) Identification of a family of zinc transporter genes from Arabidopsis that respond to zinc deficiency. Proc Natl Acad Sci USA 95:7220–7224

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Halimaa P, Lin YF, Ahonen VH, Blande D, Clemens S, Gyenesei A, Haikio E, Karenlampi SO, Laiho A, Aarts MG, Pursiheimo JP, Schat H, Schmidt H, Tuomainen MH, Tervahauta AI (2014) Gene expression differences between Noccaea caerulescens ecotypes help to identify candidate genes for metal phytoremediation. Environ Sci Technol 48:3344–3353

    Article  CAS  PubMed  Google Scholar 

  • Hammond JP, Bowen HC, White PJ, Mills V, Pyke KA, Baker AJ, Whiting SN, May ST, Broadley MR (2006) A comparison of the Thlaspi caerulescens and Thlaspi arvense shoot transcriptomes. New Phytol 170:239–260

    Article  CAS  PubMed  Google Scholar 

  • Hassinen VH, Tuomainen M, Peraniemi S, Schat H, Karenlampi SO, Tervahauta AI (2009) Metallothioneins 2 and 3 contribute to the metal-adapted phenotype but are not directly linked to Zn accumulation in the metal hyperaccumulator, Thlaspi caerulescens. J Exp Bot 60:187–196

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hegelund JN, Schiller M, Kichey T, Hansen TH, Pedas P, Husted S, Schjoerring JK (2012) Barley metallothioneins: MT3 and MT4 are localized in the grain aleurone layer and show differential zinc binding. Plant Physiol 159:1125–1137

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Calif Agric Exp Stn Circ 347:1–32

  • Hosoki R, Matsuki N, Kimura H (1997) The possible role of hydrogen sulfide as an endogenous smooth muscle relaxant in synergy with nitric oxide. Biochem Bioph Res Commun 237:527–531

    Article  CAS  Google Scholar 

  • Israr M, Jewell A, Kumar D, Sahi SV (2011) Interactive effects of lead, copper, nickel and zinc on growth, metal uptake and antioxidative metabolism of Sesbania drummondii. J Hazard Mater 186:1520–1526

    Article  CAS  PubMed  Google Scholar 

  • Jin XF, Yang XE, Islam E, Liu D, Mahmood Q, Li H, Li J (2008) Ultrastructural changes, zinc hyperaccumulation and its relation with antioxidants in two ecotypes of Sedum alfredii Hance. Plant Physio Biochem 46:997–1006

    Article  CAS  Google Scholar 

  • Khudsar T, Mahmooduzzafar IM, Sairam RK (2004) Zinc-induced changes in morpho-physiological and biochemical parameters in Artemisia annua. Biol Plantarum 48:255–260

    Article  CAS  Google Scholar 

  • Kramer U (2005) MTP1 mops up excess zinc in Arabidopsis cells. Trends Plant Sci 10:313–315

    Article  PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  • Lai D, Mao Y, Zhou H, Li F, Wu M, Zhang J, He Z, Cui W, Xie Y (2014) Endogenous hydrogen sulfide enhances salt tolerance by coupling the reestablishment of redox homeostasis and preventing salt-induced K+ loss in seedlings of Medicago sativa. Plant Sci 225:117–129

    Article  CAS  PubMed  Google Scholar 

  • Lasat MM, Pence NS, Garvin DF, Ebbs SD, Kochian LV (2000) Molecular physiology of zinc transport in the Zn hyperaccumulator Thlaspi caerulescens. J Exp Bot 51:71–79

    Article  CAS  PubMed  Google Scholar 

  • Li L, Wang Y, Shen W (2012) Roles of hydrogen sulfide and nitric oxide in the alleviation of cadmium-induced oxidative damage in alfalfa seedling roots. Biometals 25:617–631

    Article  CAS  PubMed  Google Scholar 

  • Li ZG, Yang SZ, Long WB, Yang GX, Shen ZZ (2013) Hydrogen sulphide may be a novel downstream signal molecule in nitric oxide-induced heat tolerance of maize (Zea mays L.) seedlings. Plant Cell Environ 36:1564–1572

    Article  CAS  PubMed  Google Scholar 

  • Lin YF, Aarts MG (2012) The molecular mechanism of zinc and cadmium stress response in plants. Cell Mol Life Sci 69:3187–3206

    Article  CAS  PubMed  Google Scholar 

  • Lisjak M, Teklic T, Wilson ID, Whiteman M, Hancock JT (2013) Hydrogen sulfide: environmental factor or signalling molecule? Plant Cell Environ 36:1607–1616

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Marques APGC, Oliveira RS, Rangel AOSS, Castro PML (2006) Zinc accumulation in Solanum nigrum is enhanced by different arbuscular mycorrhizal fungi. Chemosphere 65:1256–1263

    Article  CAS  PubMed  Google Scholar 

  • Mills RF, Francini A, da Rocha PSF, Baccarini PJ, Aylett M, Krijger GC, Williams LE (2005) The plant P1B-type ATPase AtHMA4 transports Zn and Cd and plays a role in detoxification of transition metals supplied at elevated levels. FEBS Lett 579:783–791

    Article  CAS  PubMed  Google Scholar 

  • Mortvedt J (2000) Bioavailability of micronutrients. In: Summer ME (ed) Handbook of soil science. CRC Press, Boca Raton, Florida, pp. 71–86

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15:473–497

  • Nevo Y, Nelson N (2006) The NRAMP family of metal-ion transporters. BBA-Mol Cell Res 1763:609–620

    CAS  Google Scholar 

  • Qiao Z, Jing T, Liu Z, Zhang L, Jin Z, Liu D, Pei Y (2015) H2S acting as a downstream signaling molecule of SA regulates Cd tolerance in Arabidopsis. Plant Soil 393:137–146

    Article  CAS  Google Scholar 

  • Remans T, Opdenakker K, Guisez Y, Carleer R, Schat H, Vangronsveld J, Cuypers A (2012) Exposure of Arabidopsis thaliana to excess Zn reveals a Zn-specific oxidative stress signature. Environ Exp Bot 84:61–71

    Article  CAS  Google Scholar 

  • Riemenschneider A, Wegele R, Schmidt A, Papenbrock J (2005) Isolation and characterization of a D-cysteine desulfhydrase protein from Arabidopsis thaliana. FEBS J 272:1291–1304

    Article  CAS  PubMed  Google Scholar 

  • Robinson NJ, Tommey AM, Kuske C, Jackson PJ (1993) Plant metallothioneins. Biochem J 295:1–10

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Sadeghzadeh B (2013) A review of zinc nutrition and plant breeding. J Soil Sci Plant Nut 13:905–927

    Google Scholar 

  • Sagardoy R, Morales F, Lopez-Millan AF, Abadia A, Abadia J (2009) Effects of zinc toxicity on sugar beet (Beta vulgaris L.) plants grown in hydroponics. Plant Biol 11:339–350

    Article  CAS  PubMed  Google Scholar 

  • Shan C, Dai H, Sun Y (2012) Hydrogen sulfide protects wheat seedlings against copper stress by regulating the ascorbate and glutathione metabolism in leaves. Aust J Crop Sci 6:248–254

    CAS  Google Scholar 

  • Shanmugam V, Lo JC, Yeh KC (2013) Control of Zn uptake in Arabidopsis halleri: a balance between Zn and Fe. Front Plant Sci 4:281

    Article  PubMed Central  PubMed  Google Scholar 

  • Shi H, Ye T, Chan Z (2014) Nitric oxide-activated hydrogen sulfide is essential for cadmium stress response in bermudagrass (Cynodon dactylon (L). Pers.). Plant Physiol Biochem 74:99–107

    Article  CAS  PubMed  Google Scholar 

  • Shi WG, Li H, Liu TX, Polle A, Peng CH, Luo ZB (2015) Exogenous abscisic acid alleviates zinc uptake and accumulation in Populus × canescens exposed to excess zinc. Plant Cell Environ 38:207–223

    Article  CAS  PubMed  Google Scholar 

  • Sinclair SA, Kramer U (2012) The zinc homeostasis network of land plants. BBA-Mol Cell Res 1823:1553–1567

  • Sinclair SA, Sherson SM, Jarvis R, Camakaris J, Cobbett CS (2007) The use of the zinc-fluorophore, Zinpyr-1, in the study of zinc homeostasis in Arabidopsis roots. New Phytol 174:39–45

    Article  CAS  PubMed  Google Scholar 

  • Song A, Li P, Li Z, Fan F, Nikolic M, Liang Y (2011) The alleviation of zinc toxicity by silicon is related to zinc transport and antioxidative reactions in rice. Plant Soil 344:319–333

    Article  CAS  Google Scholar 

  • Song Y, Hudek L, Freestone D, Puhui J, Michalczyk AA, Senlin Z, Ackland ML (2014) Comparative analyses of cadmium and zinc uptake correlated with changes in natural resistance-associated macrophage protein (NRAMP) expression in Solanum nigrum L. and Brassica Rapa. Environ Chem 11:653–660

    Article  CAS  Google Scholar 

  • Sun J, Wang RG, Zhang X, Yu YC, Zhao R, Li ZY, Chen SL (2013) Hydrogen sulfide alleviates cadmium toxicity through regulations of cadmium transport across the plasma and vacuolar membranes in Populus euphratica cells. Plant Physiol Biochem 65:67–74

    Article  CAS  PubMed  Google Scholar 

  • Teixeira J, Ferraz P, Almeida A, Verde N, Fidalgo F (2013) Metallothionein multigene family expression is differentially affected by chromium (III) and (VI) in Solanum nigrum L. plants. Food Energy Sec 2:130–140

    Article  Google Scholar 

  • Tsonev T, Lidon FJC (2012) Zinc in plants-an overview. Emir J Food Agric 24:322–333

    Google Scholar 

  • van de Mortel JE, Almar Villanueva L, Schat H, Kwekkeboom J, Coughlan S, Moerland PD, Ver Loren van Themaat E, Koornneef M, Aarts MG (2006) Large expression differences in genes for iron and zinc homeostasis, stress response, and lignin biosynthesis distinguish roots of Arabidopsis thaliana and the related metal hyperaccumulator Thlaspi caerulescens. Plant Physiol 142:1127–1147

    Article  PubMed Central  PubMed  Google Scholar 

  • Verbruggen N, Hermans C, Schat H (2009) Molecular mechanisms of metal hyperaccumulation in plants. New Phytol 181:759–776

    Article  CAS  PubMed  Google Scholar 

  • Verret F, Gravot A, Auroy P, Leonhardt N, David P, Nussaume L, Vavasseur A, Richaud P (2004) Overexpression of AtHMA4 enhances root-to-shoot translocation of zinc and cadmium and plant metal tolerance. FEBS Lett 576:306–312

    Article  CAS  PubMed  Google Scholar 

  • Wójcik M, Skórzyńska-Polit E, Tukiendorf A (2006) Organic acids accumulation and antioxidant enzyme activities in Thlaspi caerulescens under Zn and Cd stress. Plant Growth Regul 48:145–155

    Article  Google Scholar 

  • Wang BL, Shi L, Li YX, Zhang WH (2010) Boron toxicity is alleviated by hydrogen sulfide in cucumber (Cucumis sativus L.) seedlings. Planta 231:1301–1309

    Article  CAS  PubMed  Google Scholar 

  • Wang C, Zhang SH, Wang PF, Hou J, Zhang WJ, Li W, Lin ZP (2009) The effect of excess Zn on mineral nutrition and antioxidative response in rapeseed seedlings. Chemosphere 75:1468–1476

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Li L, Cui W, Xu S, Shen W, Wang R (2011) Hydrogen sulfide enhances alfalfa (Medicago sativa) tolerance against salinity during seed germination by nitric oxide pathway. Plant Soil 351:107–119

    Article  Google Scholar 

  • Weckx J, Clijsters H (1997) Zn phytotoxicity induces oxidative stress in primary leaves of Phaseolus vulgaris. Plant Physiol Biochem 35:405–410

    CAS  Google Scholar 

  • Xu J, Yin H, Li Y, Liu X (2010) Nitric oxide is associated with long-term zinc tolerance in Solanum nigrum. Plant Physiol 154:1319–1334

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Xu J, Zhu Y, Ge Q, Li Y, Sun J, Zhang Y, Liu X (2012) Comparative physiological responses of Solanum nigrum and Solanum torvum to cadmium stress. New Phytol 196:125–138

    Article  CAS  PubMed  Google Scholar 

  • Yang Z, Wu Y, Li Y, Ling HQ, Chu C (2009) OsMT1a, a type 1 metallothionein, plays the pivotal role in zinc homeostasis and drought tolerance in rice. Plant Mol Biol 70:219–229

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Hu LY, Li P, Hu KD, Jiang CX, Luo JP (2010a) Hydrogen sulfide alleviated chromium toxicity in wheat. Biol Plant 54:743–747

    Article  CAS  Google Scholar 

  • Zhang H, Hu LY, Hu KD, He YD, Wang SH, Luo JP (2008) Hydrogen sulfide promotes wheat seed germination and alleviates oxidative damage against copper stress. J Integr Plant Biol 50:1518–1529

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Tan ZQ, Hu LY, Wang SH, Luo JP, Jones RL (2010b) Hydrogen sulfide alleviates aluminum toxicity in germinating wheat seedlings. J Integr Plant Biol 52:556–567

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to Dr. Shao-Fa Zhang from Institute of Medical Plant Development in China for providing Solanum nigrum seeds kindly. This work was financially supported by the Natural Science Foundation of China (NSFC) (31570586, 31300505, 31260057 and 30930076), Research Fund of State Key Laboratory of Soil and Sustainable Agriculture, Nanjing Institute of Soil Science, Chinese Academy of Science (Y412201449) and China Postdoctoral Science Foundation (2012 M521278).

Conflict of interest

The authors declare that they have no conflict interest in this work.

Author information

Authors and Affiliations

  1. Key Laboratory for Subtropical Wetland Ecosystem Research of MOE, College of the Environment and Ecology, Xiamen University, Xiamen, Fujian, 361102, People’s Republic of China

    Xiang Liu, Juan Chen, Zhi-Jun Shen, Mei-Rong Luo, Gui-Feng Gao, Martin Simon, Kabir Ghoto & Hai-Lei Zheng

  2. College of life sciences, Xiamen University, Xiamen, Fujian, 361102, People’s Republic of China

    Guang-Hui Wang

  3. Fujian Key Laboratory of Subtropical Plant Physiology and Biochemistry, Fujian Institute of Subtropical Botany, Xiamen, Fujian, 361005, People’s Republic of China

    Wen-Hua Wang

Authors
  1. Xiang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Juan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guang-Hui Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wen-Hua Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhi-Jun Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mei-Rong Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gui-Feng Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Martin Simon
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Kabir Ghoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hai-Lei Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hai-Lei Zheng.

Additional information

Responsible Editor: Henk Schat.

Xiang Liu, Juan Chen and Guang-Hui Wang contributed equally to this work.

Electronic supplementary material

Supplementary Methods: Amplification of ZRT and MTP partial cDNAs.

ESM 1

(DOCX 4.20 mb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, X., Chen, J., Wang, GH. et al. Hydrogen sulfide alleviates zinc toxicity by reducing zinc uptake and regulating genes expression of antioxidative enzymes and metallothioneins in roots of the cadmium/zinc hyperaccumulator Solanum nigrum L.. Plant Soil 400, 177–192 (2016). https://doi.org/10.1007/s11104-015-2719-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11104-015-2719-7

Keywords

Search

Navigation