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Isolation of a cDNA clone (PcSrp) encoding serine-rich-protein from Porteresia coarctata T. and its expression in yeast and finger millet (Eleusine coracana L.) affording salt tolerance

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Abstract

A 1.4 Kb cDNA clone encoding a serine-rich protein has been isolated from the cDNA library of salt stressed roots of Porteresia coarctata, and designated as P. coarctata serine-rich-protein (PcSrp) encoding gene. Northern analysis and in situ mRNA hybridization revealed the expression of PcSrp in the salt stressed roots and rhizome of P. coarctata. However, no such expression was seen in the salt stressed leaves and in the unstressed tissues of root, rhizome and leaf, indicating that PcSrp is under the control of a salt-inducible tissue-specific promoter. In yeast, the PcSrp conferred increased NaCl tolerance, implicating its role in salinity tolerance at cellular level. Further, PcSrp was cloned downstream to rice Actin-1 promoter and introduced into finger millet through particle-inflow-gun method. Transgenic plants expressing PcSrp were able to grow to maturity and set seed under 250 mM NaCl stress. The untransformed control plants by contrast failed to survive under similar salt stress. The stressed roots of transgenic plants invariably accumulated higher Na+ and K+ ion contents compared to roots of untransformed plants; whereas, shoots of transgenics accumulated lower levels of both the ions than that of untransformed plants under identical stress, clearly suggesting the involvement of PcSrp in ion homeostasis contributing to salt tolerance.

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Abbreviations

BAP:

Benzylaminopurine

CTAB:

Cetyltrimethylammonium bromide

DEPC:

Diethylpyrocarbonate

NBT:

4-nitro blue tetrazolium chloride

PcSrp :

Porteresia coarctata serine rich protein encoding gene

PMSF:

Phenylmethyl-sulphonyl fluoride

SDS:

Sodium dodecyl sulphate

SLS:

Sodium lauryl sulphate

2,4-D:

2,4-diclorophenoxyacetic acid

References

  • Adnan AY (2004) Influence of salinity on citrus: a review paper. JCEA 5:263–272

    Google Scholar 

  • Apse MP, Aharon GS, Snedden WA, Blumwald E (1999) Salt tolerance conferred by over expression of a vacuolar Na+/H+ antiporter in Arabidopsis. Science 285:1256–1258

    Article  PubMed  CAS  Google Scholar 

  • Blom-Zandstra M, Vogelzang SA, Veen BW (1998) Sodium fluxes in sweet pepper exposed to varying sodium concentrations. J Exp Bot 49:1863–1868

    Article  CAS  Google Scholar 

  • Bohnert HJ, Shevelva E (1998) Plant stress adaptations making metabolism movement. Curr Opin Plant Biol 1:267:274

    Article  PubMed  CAS  Google Scholar 

  • Bohnert HJ, Su H, Shen B (1999) Molecular mechanisms of salinity tolerance in molecular responses to cold, drought, heat and salt stress in higher plants. In: Shinizaki EDSK, Yamaguchi-Shinizaki K (eds) R.G.Kandes company, Austin, Tx, pp 29–62

  • Bohnert HJ, Nelson DE, Jensen RG (1995) Adaptations to environmental stresses. Plant Cell 7:1099–1111

    Article  PubMed  CAS  Google Scholar 

  • Chaumont F, Barrieu F, Wojcik E, Chrispeels MJ, Jung R (2001) Aquaporins constitute a large and highly divergent protein family in maize. Plant Physiol 125:1206–1215

    Article  PubMed  CAS  Google Scholar 

  • Clarke GM, Cooke DA (1992) A basic course in statistics. Hypothesis tests using the chi-square distribution. Great Britain, Cornwall, pp 289–307

    Google Scholar 

  • Cushman JC, Bohnert HJ (2000) Genomic approaches to plant stress tolerance. Curr Opin Plant Biol 3:117–124

    Article  PubMed  CAS  Google Scholar 

  • Deutch CE, Winicov I (1995) Post-transcriptional regulation of a salt-inducible alfalfa gene encoding a putative chimeric proline-rich cell wall protein. Plant Mol Biol 27:411–418

    Article  PubMed  CAS  Google Scholar 

  • Epstein E, Rains DW (1987) Advances in salt tolerance. Plant soil 89:17–29

    Article  Google Scholar 

  • Epstein E, Norlyn JD, Rush DW, Kingsbury RW, Kelly DB, Cunningham GA, Wrona AF (1980) Saline culture of crops: a genetic approach. Science 210:399–404

    Article  PubMed  CAS  Google Scholar 

  • Gisbert C, Rus AM, Bolarin MC, Lopez-coronado JM, Arrillaga I, Montesinos C, Caro M, Serrano R, Moreno V (2000) The Yeast HAL1 gene improves salt tolerance of transgenic tomato. Plant Physiol 123:393–402

    Article  PubMed  CAS  Google Scholar 

  • Hasegawa PM, Bresson RA, Zhu JK, Bohnert HJ (2000) Plant cellular and molecular responses to high salinity. Annu Rev Plant physiol Plant Mol Biol 51:463–499

    Article  PubMed  CAS  Google Scholar 

  • Hoagland DR, Arnon DI (1938) The water culture method for growing plants without soil. Calif Exp Stn Circ 71:34–39

    Google Scholar 

  • Hong B, Uknes SJ, Ho THD (1988) Cloning and characterization of a cDNA encoding a mRNA rapidly-induced by ABA in barley aleurone layers. Plant Mol Biol 11:495–506

    Article  CAS  Google Scholar 

  • Inan G, Zhang Q, Li P, Wang Z, Caoz zhang H, Zhang C, Quist TM, Goodwin SM (2004) Salt cress: a halophyte and cryophyte Arabidopsis relative model system and its applicability to molecular genetic analysis of growth and development of extremophiles. Plant Physiol 135:1718–1737

    Article  PubMed  CAS  Google Scholar 

  • Kanhonou R, Serrano R, Palau RR (2001) A catalytic subunit of the sugar beet protein kinase CK2 is induced by salt stress and increases NaCl tolerance in Saccharomyces cerevisiae. Plant Mol Biol 47:571–579

    Article  PubMed  CAS  Google Scholar 

  • Lazof DB, Bernstein N (1999) The NaCl induced inhibition of shoot growth; the case for distributed nutrition with special consideration of calcium. Adv Bot Res 29:113–189

    Article  CAS  Google Scholar 

  • Lohaus G, Hussmann M, Pennewiss K, Schneider H, Zhu JJ, Sattelmacher B (2000) Solute balance of a maize (Zea Mays) N. source leaf as affected by salt treatment with special emphasis on phloem retranslocation and ion leaching. J Exp Bot 51:1721–1732

    Article  PubMed  CAS  Google Scholar 

  • Madhavi AL, Rao KV, Reddy VD (2005) Production of transgenic plants resistant to leaf blast disease in finger millet (Eleusine coracana (L.) Gaertn.). Plant Sci 169:657–667

    Article  CAS  Google Scholar 

  • Manoj M, Susmita M, krishnarup GD, Sitakanta P, Anirban C, Nitai C, Hait Das KP, Arun LM (2004) A Novel Salt-tolerant L-myo-Inositol-1-phosphate Synthase from Porteresia coarctata (Roxb.) Tateoka, a Halophytic Wild Rice. J Biol Chem 279:28539–28552

    Article  CAS  Google Scholar 

  • Mukhopadhyay A, Shubha V, Tyagi AK (2004) Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco. Proc Nat Acad Sci USA 101:6309–6314

    Article  PubMed  CAS  Google Scholar 

  • Munns R, Tonnet L, Shennan C, Gardner PA (1988) Effect of high external NaCl concentration on ion transport with in the shoots of Lupinus albus. II. Ions in phloem sap. Plant Cell Environ 11:291–300

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nakashima AK, Shinwani ZK, Sakuma Y, Seki M, Miura S, Shinozaki K, Yamaguchi-shinozaki K (2000) Organization and expression of two Arabidopsis DREB2 encoding genes involved in dehydration and high salt salinity responsive gene expression. Plant Mol Biol 42:657–665

    Article  PubMed  CAS  Google Scholar 

  • Pardue ML (1987) In-situ hybridization. In: Hames BD, Higgins SJ (eds) Nucleic acid hybridization: a practical approach. IRL Press, Oxford, pp 179–202

    Google Scholar 

  • Queseda V, Garcia-Martinez S, Piqueras P, Ponce MR, Micol JL (2002) Genetic Architecture of NaCl tolerance in Arabidopsis. Plant Physiol 130:951–963

    Article  CAS  Google Scholar 

  • Rohila JS, Rajender KJ, Ray Wu (2002) Genetic improvement of Basmati Rice for salt and drought tolerance by regulated expression of barley HVA1 DNA. Plant Sci 163:525–532

    Article  CAS  Google Scholar 

  • Sanan-Mishra N, Pham XH, Sudhirk S, Narendra T (2005) Pea DNA helicase 45 over expression in tobacco confers high salinity tolerance without affecting yield. Proc Nat Acad Sci USA 102:509–514

    Article  PubMed  CAS  Google Scholar 

  • Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barely: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81:8014–8018

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratories, Cold Spring Harbor

    Google Scholar 

  • Shinozaki K, Yamaguchi-shinozaki K (1997) Gene expression and signal transduction in water stress response. Plant Physiol 115:327–334

    Article  PubMed  CAS  Google Scholar 

  • Su H, Baldera SE, Vera-Estrella R, Golldack D, Quigley F, Zhao C, Pantoja O, Bohnert HJ (2003) Expression of the cation transporter McHKT1 in a halophyte. Plant Mol Biol 52:967–980

    Article  PubMed  CAS  Google Scholar 

  • Shi H, Lee B, Wu SJ, Zhu JK (2003) Overexpression of a plasma membrane Na+/H+ antiporter improves salt tolerance in Arabidopsis. Nat Biotechnol 21:81–85

    Article  PubMed  CAS  Google Scholar 

  • Taji T, Seki M, Satou M, Sakuraj T, Kobayashi M, Ishiyama K, Narusaka Y, Narusaka M, Zhu JK, Shinozaki K (2004) Comparative genomics in salt tolerance between Arabidopsis and Arabidopsis related halophyte salt cress using Arabidopsis microrray. Plant Physiol 135:1697–1709

    Article  PubMed  CAS  Google Scholar 

  • Tal M (1985) Genetics of salt tolerance in higher plants. Theoritical and practical considerations. Plant soil 89:199–226

    Article  Google Scholar 

  • Vinocur B, Altman A (2005) Recent advances in engineering plant tolerance to abiotic stress: achievements and limitations. Curr Opin Biotech 16:123–132

    Article  PubMed  CAS  Google Scholar 

  • Volkov V, Wang B, Dominy PJ, Fricke W, Amtmann A (2004) Thellungiella halophila, a salt tolerant relative of Arabidopsis-thaliana, possesses effective mechanisms to discriminate between potassium and sodium. Plant cell Environ 27:1–14

    Article  CAS  Google Scholar 

  • Winicov I, Deutch CE (1994) Characterization of a cDNA clone from salt tolerant alfalfa cells that identifies salt inducible root-specific transcripts. J Plant Physiol 144:222–228

    CAS  Google Scholar 

  • Winicov I (2000) Alfin1 transcription factor overexpression enhances plant root growth under normal and saline conditions and improves salt tolerance in alfalfa. Planta 210:416–422

    Article  PubMed  CAS  Google Scholar 

  • Winter E (1982) Salt tolerance of Trifolium alexandrium L.III. Effects of salt on ultra structure of phloem and xylem transfer cells in petioles and leaves. Aust J Plant Physiol 9:227–237

    Article  CAS  Google Scholar 

  • Wyn Jones RG, Gorham J (1986) The potential for enhancing the salt tolerance of wheat and other important crop plants. Outlook Agri 15:33–39

    Google Scholar 

  • Xiong L, Zhu JK (2002) Molecular and genetic aspects of plant responses to osmotic stress. Plant Cell Environ 25:131–139

    Article  PubMed  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. Plant Cell 6:251–264

    Article  PubMed  CAS  Google Scholar 

  • Yeo A (1998) Molecular biology of salt tolerance in the context of whole-plant physiology. J Exp Bot 49:913–929

    Article  Google Scholar 

  • Yoon HW, Kim MC, Shin PG, Kim CY, Kim JS, Lee SY, Hwang I, Bahk JD, Hong JC, Han C, Cho MJ (1997) Differential expression of two functional serine/threonine protein kinases from soyabean that have an unusual acidic domains at the carboxy terminus. Mol Gen Genet 255:359–371

    Article  PubMed  CAS  Google Scholar 

  • Zhang HX, Blumwald E (2001) Transgenic salt tolerant tomato plants accumulate salt in foliage but not in fruit. Nat Biotechnol 19:765–768

    Article  PubMed  CAS  Google Scholar 

  • Zhu JK, Hasegawa PM, Bressan RA (1997) Molecular aspects of osmotic stress in plants. Crit Rev Plant Sci 16:253-277

    Article  CAS  Google Scholar 

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Acknowledgements

We are grateful to Prof. G.M. Reddy, Founder Director, CPMB, for his constant encouragement. We offer our thanks to Prof. I. Winicov, University of Nevada, USA, for providing the probe “PA9”. We extend our thanks to Prof. P. Maruthi Mohan, Dept. of Biochemistry Osmania University for making available the facility of the Atomic Absorption Spectrophotometer. The help rendered by Mr. M.Suresh Reddy, Technical officer, CPMB, is gratefully acknowledged. Financial assistance from the Department of Biotechnology (DBT), New Delhi (India), is gratefully acknowledged. Fellowships awarded to SM and CSB by the Council of Scientific and Industrial Research (CSIR), New Delhi (India), is gratefully acknowledged.

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  1. Centre for Plant Molecular Biology, Osmania University, 500 007, Hyderabad , AP, India

    S. Mahalakshmi, G. S. B. Christopher, T. P. Reddy, K. V. Rao & V. D. Reddy

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  1. S. Mahalakshmi
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  2. G. S. B. Christopher
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  3. T. P. Reddy
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  5. V. D. Reddy
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Correspondence to V. D. Reddy.

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Mahalakshmi, S., Christopher, G.S.B., Reddy, T.P. et al. Isolation of a cDNA clone (PcSrp) encoding serine-rich-protein from Porteresia coarctata T. and its expression in yeast and finger millet (Eleusine coracana L.) affording salt tolerance. Planta 224, 347–359 (2006). https://doi.org/10.1007/s00425-005-0218-4

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  • DOI: https://doi.org/10.1007/s00425-005-0218-4

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