Skip to main content
SpringerLink
Log in

Molecular cloning and expression of chloroplast NADP-malate dehydrogenase during Crassulacean acid metabolism induction by salt stress

  • Regular Paper
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

A full-length cDNA clone for NADP+-dependent malate dehydrogenase (NADP-MDH; EC 1.1.1.82) from the facultative CAM plant,Mesembryanthemum crystallinum has been isolated and characterized. NADP-MDH is responsible for the reduction of oxaloacetate to malate in the chloroplasts of higher plants. The cDNA clone is 1747 bp in size and contains a single open reading frame encoding a 441 amino acid long precursor polypeptide with a predicted molecular weight of 47 949. The predicted, mature NADP-MDH polypeptide sequence fromM. crystallinum shares 82.7% to 84% amino acid identity with other known higher plant sequences. Genomic Southern blot analysis ofM. crystallinum DNA indicates that MDH is encoded by a small gene family. Steady-state transcript levels for chloroplast NADP-MDH decrease transiently in the leaves after salt stress and then increase to levels greater than two-fold higher than in unstressed plants. Transcript levels in roots are extremely low and are unaffected by salt-stress treatment. In vitro transcription run-on experiments using isolated nuclei from leaf tissue confirm that the accumulation of NADP-MDH transcripts is, at least in part, the result of increased transcription of this gene during salt stress. The salt-stress-induced expression pattern of this enzyme suggests that it may participate in the CO2 fixation pathway during CAM.

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

Similar content being viewed by others

Abbreviations

CAM:

Crassulacean acid metabolism

DHAP:

dihydroxyacetonephosphate

NADP-MDH:

NADP-malate dehydrogenase

NAD-MDH:

NAD-malate dehydrogenase

OAA:

oxaloacetate

PEPC:

phosphoenolpyruvate carboxylase

References

  • Buchanan BB (1980) Role of light in the regulation of chloroplast enzymes. Ann Rev Plant Physiol 31: 341–374

    Google Scholar 

  • Benton WD and Davis RW (1977) Screening lambdagt recombinant clones by hybridization to single plaquesin situ. Science 196: 180–182

    PubMed  Google Scholar 

  • Cretin C, Luchetta P, Joly C, Decottignies P, Lepiniec L, Gadal P, Sallantin M, Huet J-C and Pernollet J-C (1990) Primary structure of sorghum malate dehydrogenase (NADP) deduced from cDNA sequence. Eur J Biochem 192: 299–303

    PubMed  Google Scholar 

  • Cushman JC (1992) Characterization and expression of a NADP-Malic Enzyme cDNA induced by salt stress from the facultative CAM plant,Mesembryanthemun crystallinum. Eur J Biochem (in press)

  • Cushman JC, Meyer G, Michalowski CB, Schmitt JM and Bohnert HJ (1989) Salt stress leads to the differential expression of two isogenes of phosphoenolpyruvate carboxylase during Crassulacean acid metabolism induction in the common ice plant. Plant Cell 1: 715–725

    PubMed  Google Scholar 

  • Dale RMK, McClure BA and Houchins JP (1985) A rapid single-stranded cloning strategy for producing a sequencial series of overlapping clones for use in DNA sequencing: Application to sequencing the corn mitochondrial 18S rDNA. Plasmid 13: 31–40

    PubMed  Google Scholar 

  • Decottignies P, Schmitter J-M, Miginiac-Maslow M, LeMarechal P, Jacquot J-P and Gadal P (1988) Primary structure of the light-dependent regulatory site of corn NADP-malate dehydrogenase. J Biol Chem 263: 11780–11785

    PubMed  Google Scholar 

  • Devereux J, Haeberli P and Smithies O (1985) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12: 387–393

    Google Scholar 

  • Feinberg AP and Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13

    PubMed  Google Scholar 

  • Ferte N, Jacquot J-P and Meunier J-C (1986) Structural, immunological and kinetic comparisons of NADP dependent malate dehydrogenases from spinach (C3) and corn (C4) chloroplasts. J Biochem 154: 587–595

    Google Scholar 

  • Fickenscher K, Scheibe R and Marcus F (1987) Amino acid sequence similarity between malate dehydrogenases (NAD) and pea chloroplast malate dehydrogenase (NADP). Eur J Biochem 168: 653–658

    PubMed  Google Scholar 

  • Gupta VK and Anderson LE (1978) Light modulation of the activity of carbon metabolism enzymes in the Crassulacean acid metabolism plantKalanchoë. Plant Physiol 61: 469–471

    Google Scholar 

  • Harbinson J, Genty B and Foyer CH (1990) Relationship between photosynthetic electron transport and stromal enzyme activity in pea leaves. Plant Physiol 94: 545–553

    Google Scholar 

  • Hatch MD and Slack CR (1969) NADP-specific malate dehydrogenase and glycerate kinase in leaves and evidence for their location in chloroplasts. Biochem Biophys Res Comm 34: 589–593

    PubMed  Google Scholar 

  • Heber U (1974) Metabolite exchange between chloroplasts and cytoplasm. Ann Rev Plant Physiol 24: 393–421

    Google Scholar 

  • Higgins DG and Sharp PM (1989) Fast and sensitive multiple sequence alignments on a microcomputer. CABIOS Communications 5: 151–153

    Google Scholar 

  • Holtum JAM and Winter K (1982) Activity of enzymes of carbon metabolism during the induction of Crassulacean acid metabolism inMesembryanthemum crystallinum L. Planta 155: 8–16

    Google Scholar 

  • Hutcheson SW and Buchanan BB (1983a) Enzyme regulation in Crassulacean acid metabolism photosynthesis: Studies of the ferredoxin/thioredoxin system ofKalanchoë daigremontiana. Plant Physiol 72: 870–876

    Google Scholar 

  • Hutcheson SW and Buchanan BB (1983b) Enzyme regulation in Crassulacean acid metabolism photosynthesis. Studies on thioredoxin-linked enzymes ofKalanchoë daigremontiana. Plant Physiol 72: 877–885

    Google Scholar 

  • Ingelbrecht ILW, Herman LMF, Dekeyser RA, VanMontagu MC and Depicker AG (1989) Different 3′ end regions strongly influence the level of gene expression in plant cells. Plant Cell 1: 671–680

    PubMed  Google Scholar 

  • Joshi CP (1987) Putative polyadenylation signals in nuclear genes of higher plants: A compilation and analysis. Nucleic Acids Res 15: 9627–9640

    PubMed  Google Scholar 

  • Kagawa T and Bruno PL (1988) NADP-malate dehydrogenase from leaves ofZea mays: Purification and physical, chemical, and kinetic properties. Arch Biochem Biophys 260: 674–695

    PubMed  Google Scholar 

  • Keegstra K, Olson LJ and Theg SM (1989) Chloroplastic precursors and their transport across the envelope membranes. Annu Rev Plant Physiol 40: 471–501

    Google Scholar 

  • Kleppinger-Sparace KF, Stahl RJ and Sparace SA (1992) Energy requirements for fatty acid and glycerolipid biosynthesis from acetate by isolated pea root plastids. Plant Physiol 98: 723–727

    Google Scholar 

  • Kluge M, Fischer A and Buchanan-Bollig IC (1982) Metabolic control of CAM. In: Ting IP and Gibbs M (eds) Crassulacean Acid Metabolism, pp 31–50. Amer Soc Plant Physiol. Rockville, MD

    Google Scholar 

  • Köster S and Anderson JM (1988) The photosynthetic apparatus of C3 and CAM-inducedMesembryanthemum crystallinum L. Photosynth Res 19: 251–264

    Google Scholar 

  • Lange OL, Schulze ED, Kappen L, Evenari M and Buschbom U (1975) CO2 exchange pattern under natural conditions ofCaralluma negevensis, a CAM plant of the Negev desert. Photosynthetica 9: 318–326

    Google Scholar 

  • Littlejohn RO and Ku MSB (1984) Characterization of early morning Crassulacean acid metabolism inOpuntia erinacea var. Columbiana (Griffiths) L. Benson. Plant Physiol 74: 1050–1054

    Google Scholar 

  • Luchetta P, Cretin C and Gadal P (1990) Structure and characterization of theSorghum vulgare gene encoding NADP-malate dehydrognase. Gene 89: 171–177

    PubMed  Google Scholar 

  • Luchetta P, Cretin C and Gadal P (1991) Organization and expression of the two homologous genes encoding the NADP-malate dehydrognase inSorghum vulgare leaves. Mol Gen Genet 228: 473–481

    PubMed  Google Scholar 

  • Lütcke HA, Chow KC, Mickel FS, Moss KA, Kern HF and Scheele GA (1987) Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48

    PubMed  Google Scholar 

  • Maniatis T, Fritsch EF and Sambrook J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Metzler MC, Rothermel BA and Nelson T (1989) Maize NADP-malate deydrogenase: cDNA cloning, sequence, and mRNA characterization. Plant Mol Biol 12: 713–722

    Google Scholar 

  • Michalowski CB, Olson SW, Piepenbrock M, Schmitt JM and Bohnert HJ (1989a) Time course of mRNA induction elicited by salt stress in the common ice plant (Mesembryanthemum crystallinum). Plant Physiol 89: 811–816

    Google Scholar 

  • Michalowski CB, Schmitt JM and Bohnert HJ (1989b) Expression during salt stress and nucleotide sequence of cDNA for ferredoxin-NADP+ reductase fromMesembryanthemum crystallinum. Plant Physiol 89: 817–822

    Google Scholar 

  • Michalowski CB, DeRocher EJ, Bohnert HJ and Salvucci ME (1992) Phosphoribuloskinase from ice plant: Transcription, transcripts and proteins expression during environmental stress. Photosynth Res 31: 127–138

    Google Scholar 

  • Nakamoto H and Edwards GE (1983) Dark activation of maize leaf NADP-malate dehydrogenase and pyruvate, orthophosphate dikinase in vivo under anaerobic conditions. Plant Sci Lett 32: 139–146

    Google Scholar 

  • Osmond CB (1984) CAM: Regulated photosynthetic metabolism for all seasons. In: Sybesma C (ed) Advances in Photosynthesis Research, Vol III, pp 557–564. Martinus Nijhoff/Dr W. Junk publishers, The Hague

    Google Scholar 

  • Osmond CB and Holtum JAM (1981) Crassulacean acid metabolism. In: Hatch MD and Boardman NK (eds) The Biochemistry of Plants: A Comprehensive Treatise, Vol 8, pp 283–328. Academic Press, London/New York

    Google Scholar 

  • Osmond CB, Nott DL and Firth PM (1979) Carbon assimilation patterns and growth of the introduced CAM plantOpuntia inermis in Eastern Australia. Oecologia 40: 331–350

    Google Scholar 

  • Ostrem JA, Olson SW, Schmitt JM and Bohnert HJ (1987) Salt stress increases the level of translatable mRNA for Phosphoenolpyruvate carboxylase inMesembryanthemum crystallinum. Plant Physiol 84: 1270–1275

    Google Scholar 

  • Pearson WR and Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85: 2444–2448

    PubMed  Google Scholar 

  • Ray TB and Black CC (1979) The C4 pathway and its regulation. In: Gibbs M and Latzko E (eds) Encyclopedia of Plant Physiology, New Series Vol 6, pp 77–112. Springer-Verlag, New York

    Google Scholar 

  • Scheibe R (1987) NADP+-malate dehydrogenase in C3-plants: Regulation and role of a light-activated enzyme. Physiol Plant 71: 393–400

    Google Scholar 

  • Scheibe R, Kampfenkel K, Wessels R and Tripier D (1991) Primary structure and analysis of the location of the regulatory disulfide bond of pea chloroplast NADP-malate dehydrogenase. Biochem Biophys Acta 1076: 1–8

    PubMed  Google Scholar 

  • Schuber M, Kluge M (1981) In situ studies on Crassulacean acid metabolism inSedum acre L. andSedum mite Gil Oecologia 50: 82–87

    Google Scholar 

  • Steinmüller K and Apel K (1986) A simple and efficient procedure for isolating chromatin which is suitable for studies of DNase I-sensitive and hypersensitive sites. Plant Molec Biol 7: 87–94

    Google Scholar 

  • Tabor S and Richardson CC (1987) DNA sequence analysis with a modified bacteriophage T7 DNA polymerase Proc Natl Acad Sci USA 84: 4767–4771

    PubMed  Google Scholar 

  • Vieira J and Messing J (1987) Production of single-stranded plasmid DNA. Meth Enzymol 153: 3–12

    PubMed  Google Scholar 

  • Vivkanandan M and Edwards GE (1987) Light and dark anaerobic activation of NADP-malate dehydrogenase in pea leaves and chloroplasts. In: Biggins J (ed) Progress in Photosynthesis Research, Vol III, pp 257–260. Martinus Nijhoff Publishers, Dordrecht

    Google Scholar 

  • Werdan K, Heldt HW and Milovancev M (1975) The role of pH in the regulation of carbon fixation in the chloroplast stroma. Studies on CO2 fixation in the light and dark. Biochim Biophys Acta 396: 276–292.

    PubMed  Google Scholar 

  • Winter K (1980) Carbon dioxide and water vapor exchange in the Crassulacean acid metabolism plantKalanchoë pinnata during a prolonged light period. Plant Physiol 66: 917–921

    Google Scholar 

  • Winter K (1985) Crassulacean acid metabolism. In: Barber J and Baker NR (eds) Photosynthatic Mechanisms and the Environment, pp 329–387. Elsevier, Amsterdam

    Google Scholar 

  • Winter K and Tenhunen JD (1982) Light-stimulated burst of carbon dioxide uptake following nocturnal acidification in the Crassulacean acid metabolism plantKalanchoë daigremontiana. Plant Physiol 70: 1718–1722

    Google Scholar 

  • Winter K, Foster JG, Edwards GE and Holtum JAM (1982) Intracellular localization of enzymes of carbon metabolism inMesembryanthemum crystallinum exhibiting C3 photosynthetic characteristics or performing Crassulacean acid metabolism. Plant Physiol 69: 300–307

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, University of Arizona, 85721, Tucson, AZ, USA

    John C. Cushman

Authors
  1. John C. Cushman
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The nucleotide sequence data reported in this paper will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession number X63727.

The nucleotide sequence data reported in this paper will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession number X63727.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cushman, J.C. Molecular cloning and expression of chloroplast NADP-malate dehydrogenase during Crassulacean acid metabolism induction by salt stress. Photosynth Res 35, 15–27 (1993). https://doi.org/10.1007/BF02185408

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02185408

Key words

Search

Navigation