Plant Molecular Biology

, Volume 38, Issue 6, pp 1089–1099 | Cite as

Desiccation- and abscisic acid-responsive genes encoding major intrinsic proteins (MIPs) from the resurrection plant Craterostigma plantagineum

  • Jean-Baptiste Mariaux
  • Christine Bockel
  • Francesco Salamini
  • Dorothea Bartels


Major intrinsic proteins (MIPs) are a family of channel proteins that are mainly represented by aquaporins in plants. These are divided into TIPs (tonoplast intrinsic proteins) and PIPs (plasma membrane intrinsic proteins) according to their subcellular localization. Homologues to PIPs and TIPs were isolated from the desiccation-tolerant resurrection plant Craterostigma plantagineum by two approaches: firstly, a cDNA library constructed from RNA of dehydrated C. plantagineum leaves was screened with an Arabidopsis thaliana Ath-PIP1b cDNA probe and, secondly, a cDNA library was screened differentially to isolate early drought-induced transcripts. According to sequence homologies the isolated cDNA clones were grouped as follows: Cp-PIPa, Cp-PIPb, Cp-PIPc and Cp-TIP.

Cp-PIPa, Cp-PIPc and Cp-TIP transcript accumulation was regulated by dehydration and abscisic acid (ABA). Within the Cp-PIPa group transcripts were regulated either by drought only or by drought and ABA, indicating that ABA-dependent and -independent signal transduction pathways lead to Cp-PIPa expression. Comparison of Cp-PIPa expression in detached leaves and in whole plants suggested the involvement of a signal transmitted in the whole plant in response to drought. Cp-PIPb transcript levels were constitutive in all organs tested.

Antibodies raised against a Cp-PIPA protein recognized a polypeptide with an apparent molecular mass of 28 kDa. Using these antibodies it was shown that both Cp-PIPA and Cp-PIPB proteins were localized to the plasma membrane. The role of different members of the MIP group in the dehydration response is discussed.

abscisic acid aquaporin Craterostigma plantagineum drought plasma membrane intrinsic proteins (PIP) tonoplast intrinsic protein (TIP) 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Agre P, Brown D, Nielsen F: Aquaporin water channels: Unanswered questions and unresolved controversies. Curr Opin Cell Biol 7: 472–483 (1995).Google Scholar
  2. 2.
    Bartels D, Schneider K, Terstappen g, Piatkowski D, Salamini F: Molecular cloning of abscisic acid-modulated genes which are induced during desiccation of the resurrection plant Craterostigma plantagineum. Planta 181: 27–34 (1990).Google Scholar
  3. 3.
    Bernacchia g, Salamini F, Bartels D: Molecular characterisation of the rehydration process in the resurrection plant Craterostigma plantagineum. Plant Physiol 111: 1043–1050 (1996).Google Scholar
  4. 4.
    Bianchi G, Gamba A, Murelli C, Salamini F, Bartels D: Low molecular weight solutes in desiccated and ABA-treated calli and leaves of Craterostigma plantagineum. Phytochemistry 31: 1917–1922 (1992).Google Scholar
  5. 5.
    Bockel C, Salamini F, Bartels D: Isolation and characterization of genes expressed during early events of the dehydration process in the resurrection plant Craterostigma plantagineum. J Plant Physiol, in press.Google Scholar
  6. 6.
    Bohnert HJ, Nelson DE, Jensen RG: Adaptations to environmental stresses. Plant Cell 7: 1099–1111 (1995).Google Scholar
  7. 7.
    Chrispeels MJ, Maurel C: Aquaporins: The molecular basis of facilitated water movement through living plant cells? Plant Physiol 105: 9–13 (1994).Google Scholar
  8. 8.
    Coligan JE, Dunn BM, Ploegh HL, Speicher DW, Wingfield PT: Current Protocols in Protein Science, vol I. John Wiley, New York (1995).Google Scholar
  9. 9.
    Daniels MJ, Mirkov TE, Chrispeels MJ: The plasma membrane of Arabidopsis thaliana contains a mercury-insensitive aquaporin that is a homologue of the tonoplast water channel protein TIP. Plant Physiol 106: 1325–1333 (1994).Google Scholar
  10. 10.
    Daniels MJ, Chaumont F, Mirkov TE, Chrispeels MJ: Characterization of a new vacuolar membrane aquaporin sensitive to mercury at a unique site. Plant Cell 8: 587–599 (1996).Google Scholar
  11. 11.
    Feng DF, Doolittle RF: Progressive sequence alignment as a prerequisite to correct phylogenetic trees. J Mol Evol 25: 351–360 (1987).Google Scholar
  12. 12.
    Fray RG, Wallace A, Grierson D, Lycett GW: Nucleotide sequence and expression of a ripening and water stress-related cDNA from tomato with homology to the MIP class of membrane channel proteins. Plant Mol Biol 24: 539–543 (1994).Google Scholar
  13. 13.
    Furini A, Koncz C, Salamini F, Bartels D: High level transcription of a member of a repeated gene family confers dehydration tolerance to callus tissues of Craterostigma plantagineum. EMBO J 16: 3599–3608 (1997).Google Scholar
  14. 14.
    Gatenby AA, Castleton JA, Saul MW: Expression in E. coli of maize and wheat chloroplast genes for the large subunit of ribulose biphosphate carboxylase. Nature 291: 117–121 (1981).Google Scholar
  15. 15.
    Gerlach WL, Bedbrook JR: Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucl Acids Res 7: 1869–1885 (1979).Google Scholar
  16. 16.
    Glass DB, El-Maghrabi MR, Pilkis SJ: Synthetic peptides corresponding to the site phosphorylated in 6-phosphofructo-2-kinase/fructose-2,6-biphosph atase as substrates of cyclic nucleotide-dependent protein kinases. J Biol Chem 261: 2987–2993 (1986).Google Scholar
  17. 17.
    Guerrero FD, Crossland L: Tissue-specific expression of a plant turgor-responsive gene with amino acid sequence homology to transport-facilitating proteins. Plant Mol Biol 21: 929–935 (1993).Google Scholar
  18. 18.
    Hartmann-Bouillon MA, Benveniste P: Sterol biosynthetic capability of purified membrane fractions from maize coleoptiles. Phytochemistry 17: 1037–1042 (1978).Google Scholar
  19. 19.
    Höfte H, Hubbard L, Reizer J, Ludevid D, Herman EM, Chrispeels M: Vegetative and seed-specific forms of tonoplast intrinsic protein in the vacuolar membrane of Arabidopsis thaliana. Plant Physiol 99: 561–570 (1992).Google Scholar
  20. 20.
    Ingram J, Bartels D: The molecular basis of dehydration tolerance in plants. Annu Rev Plant Physiol Plant Mol Biol 47: 377–403 (1996).Google Scholar
  21. 21.
    Jochem P, Lüttge U: Proton translocating enzymes at the tonoplast of leaf cells of the CAM plant Kalanchoë daigremontiana. I. The ATPase. J Plant Physiol 129: 251–268 (1987).Google Scholar
  22. 22.
    Johansson I, Larsson C, Ek B, Kjellbom P: The major integral proteins of spinach leaf plasma membranes are putative aquaporins and are phosphorylated in response to Ca2+ and apoplastic water potential. Plant Cell 8: 1181–1191 (1996).Google Scholar
  23. 23.
    Kaldenhoff R, Kölling A, Richter G: A novel blue lightand abscisic acid-inducible gene of Arabidopsis thaliana encoding an intrinsic membrane protein. Plant Mol Biol 23: 1187–1198 (1993).Google Scholar
  24. 24.
    Kaldenhoff R, Kölling A, Meyers J, Karmann U, Ruppel G, Richter G: The blue light-responsive Ath-H2 gene of Arabidopsis thaliana is primarily expressed in expanding as well as in differentiating cells and encodes a putative channel protein of the plasmalemma. Plant J 7: 87–95 (1995).Google Scholar
  25. 25.
    Kaldenhoff R, Kölling A, Richter G: Regulation of the Arabidopsis thaliana aquaporin gene Ath-H2 (PIP1b). J Photochem Photobiol 36: 351–354 (1996).Google Scholar
  26. 26.
    Kammerloher W, Fischer U, Piechottka GP, Schäffner AR: Water channels in the plant plasma membrane cloned by immunoselection from a mammalian expression system. Plant J 6: 187–199 (1994).Google Scholar
  27. 27.
    Klein P, Kanehisa M, DeLisi C: The detection-and classification of membrane-spanning domains. Biochim Biophys Acta 815: 468–476 (1985).Google Scholar
  28. 28.
    Laemmli UK: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227: 680–685 (1970).Google Scholar
  29. 29.
    Lee JW, Zhang Y, Shomer NH, Louis CF, Roberts DM: Phosphorylation of nodulin 26 on serine 262 affects its voltagesensitive channel activity in planar lipid bilayers. J Biol Chem 270: 27051–27057 (1995).Google Scholar
  30. 30.
    Liu Q, Umeda M, Uchimiya H: Isolation and expression analysis of two rice genes encoding the major intrinsic protein. Plant Mol Biol 26: 2003–2007 (1994).Google Scholar
  31. 31.
    Ludevid D, Höfte H, Himelblau E, Chrispeels MJ: The expression pattern of the tonoplast intrinsic protein γ-TIP in Arabidopsis thaliana is correlated with cell enlargement. Plant Physiol 100: 1633–1699 (1992).Google Scholar
  32. 32.
    Marquardt G, Lüttge U: Proton translocating enzymes at the tonoplast of leaf cells of the CAM plant Kalanchoë daigremontiana. II. The Pyrophosphatase. J Plant Physiol 129: 269–286 (1987).Google Scholar
  33. 33.
    Maurel C: Aquaporins and water permeability of plant membranes. Annu. Rev. Plant Physiol Plant Mol Biol 48: 399–429 (1997).Google Scholar
  34. 34.
    Maurel C, Kado RT, Guern J, Chrispeels MJ: Phosphorylation regulates the water channel activity of the seed-specific aquaporin α-TIP. EMBO J 14: 3028–3035 (1995).Google Scholar
  35. 35.
    Ouyang LY, Whelan J, Weaver CD, Roberts DM, Day DA: Protein phosphorylation stimulates the rate of malate uptake across the peribacteroid membrane of soybean nodules. FEBS Lett 293: 188–190 (1991).Google Scholar
  36. 36.
    Palmgren MG, Askerlund P, Fredrikson K, Widell S, Sommarin M, Larsson C: Sealed inside-out and right-side-out plasma membrane vesicles. Plant Physiol 92: 871–880 (1990).Google Scholar
  37. 37.
    Pearson WR, Lipman DJ: Improved tools for biological sequence analysis. Proc. Natl Acad Sci USA 85: 2444–2448 (1988).Google Scholar
  38. 38.
    Piatkowski D, Schneider K, Salamini F, Bartels D: Characterization of five abscisic acid-responsive cDNA clones isolated from the desiccation-tolerant plant Craterostigma plantagineum and their relationship to other water-stress genes. Plant Physiol 94: 1682–1688 (1990).Google Scholar
  39. 39.
    Pinna LA: Casein kinase 2: a ‘eminence grise’ in cellular regulation? Biochem Biophys Acta 1054: 267–284 (1990).Google Scholar
  40. 40.
    Quail PH: plant cell fractionation. Annu Rev Plant Physiol 30: 425–485 (1979).Google Scholar
  41. 41.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  42. 42.
    Schäffner AR: Aquaporin function, structure and expression: are there more surprises to surface in water relations? Planta 204: 131–139 (1998).Google Scholar
  43. 43.
    Shagan T, Bar-Zvi D: Nucleotide sequence of an Arabidopsis thaliana turgor-responsive cDNA clone encoding TMP-A, a transmembrane protein containing the major intrinsic protein motif. Plant Physiol 101: 1397–1398 (1993).Google Scholar
  44. 44.
    Shagan T, Meraro D, Bar-Zvi D: Nucleotide sequence of an Arabidopsis thaliana turgor-responsive TMP-B cDNA clone encoding a transmembrane protein with a major intrinsic protein motif. Plant Physiol 102: 689–690 (1993).Google Scholar
  45. 45.
    Schmidt J, J ohn M, Wieneke U, Krüssmann HD, Schell J: Expression of the nodulation gene nodA in Rhizobium meliloti and localisation of the gene in the cytosol. Proc Natl Acad Sci USA 83: 9581–9585 (1986).Google Scholar
  46. 46.
    Weig A, Deswarte C, Chrispeels MJ: The major intrinsic protein family of Arabidopsis has 23 members that form three distinct groups with functional aquaporins in each group. Plant Physiol. 114: 1347–1357 (1997).Google Scholar
  47. 47.
    Woodgett JR, Gould KL, Hunter T: Substrate specificity of protein kinase C. Use of synthetic peptides corresponding to physiological sites as probes for substrate recognition requirements. Eur J Biochem 161: 177–184 (1986).Google Scholar
  48. 48.
    Yamada S, Katsuhara M, Kelly WB, Michalowski CB, Bohnert HJ: A family of transcripts encoding water channel proteins: tissue-specific expression in the common ice plant. Plant Cell 7: 1129–1142 (1995).Google Scholar
  49. 49.
    Yamaguchi-Shinozaki K, Koizumi M, Urao S, Shinozaki K: Molecular cloning and characterization of 9 cDNAs for genes that are responsive to desiccation in Arabidopsis thaliana: sequence analysis of one cDNA clone that encodes a putative transmembrane channel protein. Plant Cell Physiol 33: 217–224 (1992).Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Jean-Baptiste Mariaux
    • 1
  • Christine Bockel
    • 1
  • Francesco Salamini
    • 1
  • Dorothea Bartels
    • 1
  1. 1.Max-Planck-Institut für Züchtungsforschung, Carl-von-Linné-Weg 10KölnGermany

Personalised recommendations