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Expression of a Suaeda salsa Vacuolar H+/Ca2+ Transporter Gene in Arabidopsis Contributes to Physiological Changes in Salinity

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Abstract

A cDNA (SsCAX1) encoding a tonoplast-localised Ca2+/H+ exchanger was isolated from a C3 halophyte Suaeda salsa (L.). To clarify the role of SsCAX1 in plant salt tolerance, Arabidopsis plants expressing SsCAX1 were treated with NaCl. Transgenic Arabidopsis plants displayed decreased salt tolerance. Although Na+ content was close to wild-type plants, transgenic plants accumulated more Ca2+ and retained less K+ in leaves than the wild-type plants in salinity. Furthermore, transgenic lines held higher leaf membrane leakage than wild-type lines under NaCl treatment. In addition, transgenic plants showed a 23% increase in vacuolar H+-ATPase activity compared with wild-type plants in normal condition. But the leaf V-H+-ATPase activity had subtle changes in transgenic plants, while significantly increased in wild-type plants under saline condition. These results suggested that regulated expression of Ca2+/H+ antiport was critical for maintenance of cation homeostasis and activity of V-H+-ATPase under saline condition.

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Abbreviations

CAX:

Cation exchanger

DTT:

Dithiothreitol

EGTA:

Ethylene glycol bis(β-aminoethyl) tetraacetic acid

HEPES:

N-2-Hydroxyethyl-piperazine-N′-2-ethanesulfonic acid

MES:

2-(N-Morpholine) ethanesulfonic acid

PMSF:

Phenylmethyl sulfonyl fluoride

PVP:

Polyvinylpyrrolidone

SsCAX1:

Suaeda salsa cation exchanger

TL:

Transgenic lines

Tricine:

N-tris-(Hydroxymethyl)-methylglycine

Tris:

Tris(hydroxymethyl) aminomethane

WT:

Wild type

References

  • Barkla BJ, Hirschi KD, Pittman JK (2008) Exchangers man the pumps: functional interplay between proton pumps and proton-coupled Ca2+ exchangers. Plant Signal Behav 3:354–356

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Cheng NH, Pittman JK, Barkla BJ, Shigaki T, Hirschi KD (2003) The Arabidopsis cax1 mutant exhibits impaired ion homeostasis, development, and hormonal responses, and reveals interplay among vacuolar transporters. Plant Cell 15:347–364

    Article  PubMed  CAS  Google Scholar 

  • Cheng NH, Pittman JK, Shigaki T, Hirschi KD (2002) Characterization of CAX4, an Arabidopsis H+/cation antiporter. Plant Physiol 128:1245–1254

    Article  PubMed  CAS  Google Scholar 

  • Chomczynski P, Sacci N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 162:156–159

    Article  PubMed  CAS  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  PubMed  CAS  Google Scholar 

  • Das R, Pandey GK (2010) Expressional analysis and role of calcium regulated kinases in abiotic stress signaling. Curr Genomics 11:2–13

    Article  PubMed  CAS  Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21

    Article  CAS  Google Scholar 

  • Geisler M, Frangne N, Gomès E, Martinoia E, Palmgren MG (2000) The ACA4 gene of Arabidopsis encodes a vacuolar membrane calcium pump that improves salt tolerance in yeast. Plant Physiol 124:1814–1827

    Article  PubMed  CAS  Google Scholar 

  • Han N, Shao Q, Bao HY, Wang BS (2011) Cloning and characterization of a Ca2+/H+ antiporter from halophyte Suaeda salsa L. Plant Mol Biol Rep 29:449–457

    Article  CAS  Google Scholar 

  • Hetherington AM, Brownlee C (2004) The generation of Ca2+ signals in plants. Annu Rev Plant Physiol Plant Mol Biol 55:401–427

    Article  CAS  Google Scholar 

  • Hirschi KD (1999) Expression of Arabidopsis CAX1 in tobacco: altered calcium homeostasis and increased stress sensitivity. Plant Cell 11:2113–2122

    Article  PubMed  CAS  Google Scholar 

  • Hirschi KD, Miranda ML, Wilganowski NL (2001) Phenotypic changes in Arabidopsis caused by expression of a yeast vacuolar Ca2+/H+ antiporter. Plant Mol Biol 46:57–65

    Article  PubMed  CAS  Google Scholar 

  • Hirschi KD, Zhen RG, Cunningham KW, Rea PA, Fink GR (1996) CAX1, an H+/Ca2+ antiporter from Arabidopsis. Proc Natl Acad Sci 93:8782–8786

    Article  PubMed  CAS  Google Scholar 

  • Kamiya T, Akahori T, Maeshima M (2005) Expression profile of the genes for rice cation/H+ exchanger family and functional analysis in yeast. Plant Cell Physiol 46:1735–1740

    Article  PubMed  CAS  Google Scholar 

  • Kamiya T, Maeshima M (2004) Residues in internal repeats of the rice cation/H+ exchanger are involved in the transport and selection of cations. J Biol Chem 279:812–819

    Article  PubMed  CAS  Google Scholar 

  • Knight H, Knight MR (2000) Imaging spatial and cellular characteristics of low temperature calcium signature after cold acclimation in Arabidopsis. J Exp Bot 51:1679–1686

    Article  PubMed  CAS  Google Scholar 

  • Krebs M, Beyhl D, Görlicha E, Al-Rasheidc K, Martenb I, Stierhof YD, Hedrichb R, Schumacher K (2010) Arabidopsis V-ATPase activity at the tonoplast is required for efficient nutrient storage but not for sodium accumulation. Proc Natl Acad Sci USA 107:3251–3256

    Article  PubMed  CAS  Google Scholar 

  • Kudla J, Batistič O, Hashimoto K (2010) Calcium signals: the lead currency of plant information processing. Plant Cell 22:541–563

    Article  PubMed  CAS  Google Scholar 

  • Li L, Kim BG, Cheong YH, Pandey GK, Luan SA (2006) Ca2+ signaling pathway regulates a K+ channel for low-K response in Arabidopsis. Proc Natl Acad Sci USA 103:12625–126530

    Article  PubMed  CAS  Google Scholar 

  • Li W, Wang D, Jin T, Chang Q, Yin D, Xu S, Liu B, Liu L (2011) The vacuolar Na+/H+ antiporter gene SsNHX1 from the halophyte Salsola soda confers salt tolerance in transgenic alfalfa (Medicago sativa L.). Plant Mol Biol Rep 29:278–290

    Article  CAS  Google Scholar 

  • Lin T, Morales MF (1977) Application of a one step procedure for measuring inorganic phosphate in the presence of proteins: the actomyosin ATPase system. Anal Biochem 77:10–17

    Article  PubMed  CAS  Google Scholar 

  • Liu J, Zhu JK (1998) A calcium sensor homolog required for plant salt tolerance. Science 280:1943–1945

    Article  PubMed  CAS  Google Scholar 

  • Mandala S, Taiz L (1985) Partial purification of a tonoplast ATPase from corn coleoptiles. Plant Physiol 78:327–333

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Pittman JK, Edmond C, Sunderland PA, Bray CM (2009) A cation-regulated and proton gradient-dependent cation transporter from Chlamydomonas reinhardtii has a role in calcium and sodium homeostasis. J Biol Chem 284:525–533

    Article  PubMed  CAS  Google Scholar 

  • Pittman JK, Hirschi KD (2001) Regulation of CAX1, an Arabidopsis Ca2+/H+ antiporter: identification of an N-terminal autoinhibitory domain. Plant Physiol 127:1020–1029

    Article  PubMed  CAS  Google Scholar 

  • Pittman JK, Shigaki T, Marshall JL, Morris JL, Cheng NH, Hirschi KD (2004) Functional and regulatory analysis of the Arabidopsis thaliana CAX2 cation transporter. Plant Mol Biol 56:959–971

    Article  PubMed  CAS  Google Scholar 

  • Qiu NW, Chen M, Guo JR, Ma XL, Wang BS (2007) Coordinate up-regulation of V-H+-ATPase and vacuolar Na+/H+ antiporter as a response to NaCl treatment in a C3 halophyte Suaeda salsa. Plant Sci 172:1218–1225

    Article  CAS  Google Scholar 

  • Ranf S, Wünnenberg P, Lee J, Becker D, Dunkel M, Hedrich R, Scheel D, Dietrich P (2008) Loss of the vacuolar cation channel, AtTPC1, does not impair Ca2+ signals induced by abiotic and biotic stresses. Plant J 53:287–299

    Article  PubMed  CAS  Google Scholar 

  • Reddy ASN, Ali GS, Celesnik H, Day IS (2011) Coping with stresses: roles of calcium- and calcium/calmodulin-regulated gene expression. Plant Cell 23:2010–2032

    Article  PubMed  CAS  Google Scholar 

  • Saltveit ME (2002) The rate of ion leakage from chilling-sensitive tissue does not immediately increase upon exposure to chilling temperatures. Postharvest Biol Technol 26:295–304

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual, 2nd edn. Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Shigaki T, Pittman JK, Hirschi KD (2003) Manganese specificity determinants in the Arabidopsis metal/H+ antiporter CAX2. J Biol Chem 278:6610–6617

    Article  PubMed  CAS  Google Scholar 

  • Shigaki T, Rees I, Nakhleh L, Hirschi KD (2006) Identification of three distinct phylogenetic groups of CAX cation/proton antiporters. J Mol Evol 63:815–825

    Article  PubMed  CAS  Google Scholar 

  • Sze H, Li X, Palmgren MG (1999) Energization of plant cell membranes by H+-pumping ATPase: regulation and biosynthesis. Plant Cell 11:677–689

    Article  PubMed  CAS  Google Scholar 

  • Tang R, Li C, Xu K, Du Y, Xia T (2010) Isolation, functional characterization, and expression pattern of a vacuolar Na+/H+ antiporter gene TrNHX1 from Trifolium repens L. Plant Mol Biol Rep 28:102–111

    Article  CAS  Google Scholar 

  • Wang BS, Lüttge U, Ratajczak R (2001) Effect of salt treatment and osmotic stress on V-ATPase and V-PPase in leaves of the halophyte Suaeda salsa. J Exp Bot 52:2355–2365

    Article  PubMed  CAS  Google Scholar 

  • Xu C, Zheng L, Gao C, Wang C, Liu G, Jing J, Wang Y (2011) Ovexpression of a vacuolar H+-ATPase c subunit gene mediates physiological changes leading to enhanced salt tolerance in transgenic tobacco. Plant Mol Biol Rep 29:424–430

    Article  CAS  Google Scholar 

  • Xu J, Li HD, Chen LQ, Wang Y, Liu LL, He L, Wu W (2006) A protein kinase, interacting with two calcineurin B-like proteins, regulates K+ transporter AKT1 in Arabidopsis. Cell 125:1347–1360

    Article  PubMed  CAS  Google Scholar 

  • Yang G, Zhou R, Tang T, Chen X, Ouyang J, He L, Li W, Chen S, Guo M, Li X, Zhong C, Shi S (2011) Gene expression profiles in response to salt stress in Hibiscus tiliaceus. Plant Mol Biol Rep 29:609–617

    Article  Google Scholar 

  • Yang MF, Song J, Wang BS (2010) Organ-specific responses of vacuolar H + -ATPase in the shoots and roots of C3 halophyte Suaeda salsa to NaCl. J Integr Plant Biol 52:308–314

    Article  PubMed  CAS  Google Scholar 

  • Zhao J, Connorton JM, Guo Y, Li X, Shigaki T, Hirschi KD, Pittman JK (2009) Functional studies of split Arabidopsis Ca2+/H+ exchangers. J Biol Chem 284:34075–34083

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful for the financial support from the NSFC (National Natural Science Research Foundation of China, project No. 30670177) and the Doctoral Foundation of Shandong Province (BS2009NY035).

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Authors and Affiliations

  1. Key Laboratory of Biotechnology, College of Food and Biologic Engineering, Shandong Polytechnic University, Daxue Road, Western University Science Park, Jinan, 250353, China

    Ning Han, Wenjun Lan, Xi He & Xinjie Zhao

  2. Key Laboratory of Plant Stress Research, College of Life Science, Shandong Normal University, Jinan, 250014, China

    Qun Shao & Baoshan Wang

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  1. Ning Han
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  2. Wenjun Lan
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  3. Xi He
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  4. Qun Shao
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  5. Baoshan Wang
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  6. Xinjie Zhao
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Correspondence to Xinjie Zhao.

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Han, N., Lan, W., He, X. et al. Expression of a Suaeda salsa Vacuolar H+/Ca2+ Transporter Gene in Arabidopsis Contributes to Physiological Changes in Salinity. Plant Mol Biol Rep 30, 470–477 (2012). https://doi.org/10.1007/s11105-011-0353-y

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