Plant Molecular Biology

, Volume 62, Issue 1–2, pp 305–323 | Cite as

Localization and Quantification of Plasma Membrane Aquaporin Expression in Maize Primary Root: A Clue to Understanding their Role as Cellular Plumbers

  • Charles Hachez
  • Menachem Moshelion
  • Enric Zelazny
  • Damien Cavez
  • François Chaumont
Original Paper


Water movement across root tissues occurs by parallel apoplastic, symplastic, and transcellular pathways that the plant can control to a certain extent. Because water channels or aquaporins (AQPs) play an important role in regulating water flow, studies on AQP mRNA and protein expression in different root tissues are essential. Here, we quantified and localized the expression of Zea mays plasma membrane AQPs (ZmPIPs) in primary root tip using in situ and quantitative RT-PCR and immunodetection approaches. All ZmPIP genes except ZmPIP2;7 were expressed in primary roots. Expression was found to be dependent on the developmental stage of the root, with, in general, an increase in expression towards the elongation and mature zones. Two genes, ZmPIP1;5 and ZmPIP2;5, showed the greatest increase in expression (up to 11- and 17-fold, respectively) in the mature zone, where they accounted for 50% of the total expressed ZmPIPs. The immunocytochemical localization of ZmPIP2;1 and ZmPIP2;5 in the exodermis and endodermis indicated that they are involved in root radial water movement. In addition, we detected a polar localization of ZmPIP2;5 to the external periclinal side of epidermal cells in root apices, suggesting an important role in water uptake from the root surface. Finally, protoplast swelling assays showed that root cells display a variable, but globally low, osmotic water permeability coefficient (P f < 10 µm/s). However, the presence of a population of cells with a higher P f (up to 26 µm/s) in mature zone of the root might be correlated with the increased expression of several ZmPIP genes.


Aquaporin PIP mRNA and protein expression Root water movement 



abscisic acid




root hydraulic conductivity


osmotic water permeability coefficient


reverse transcription-PCR


Zea mays plasma membrane intrinsic protein


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



This work was supported by grants from the Belgian National Fund for Scientific Research (FNRS), the Interuniversity Attraction Poles Programme—Belgian Science Policy, and the “Communauté française de Belgique—Actions de Recherches Concertées”. F.C. is a Senior Research Associate and C.H. a Research Fellow at the FNRS; E.Z. is a Research Fellow at the Fonds pour la Formation à la Recherche dans l’Industrie et dans l’Agriculture. We thank R. Jung (Pioneer Hi-Bred International) for providing ZmPIP cDNAs, M. Maeshima (Nagoya University, Japan) and M. Boutry (Université catholique de Louvain) for supplying the anti-RsPIP1 and anti-PMA antibodies, respectively, and T. Trombik and E. Peeters (Université catholique de Louvain) for supplying pGex-KG’ plasmid. We are very grateful to X. Draye and T. Lavigne for the use of the aeroponics facility and advices, and to M. Boutry and X. Draye for their critical reading of the manuscript.

Supplementary material

11103_2006_Article_9022_MOESM1_ESM.doc (154 kb)
Tables (DOC 154 KB)


  1. Agre P, Kozono D (2003) Aquaporin water channels: molecular mechanisms for human diseases. FEBS Lett 555:72–78CrossRefPubMedGoogle Scholar
  2. Alexandersson E, Fraysse L, Sjovall-Larsen S, Gustavsson S, Fellert M, Karlsson M, Johanson U, Kjellbom P (2005) Whole gene family expression and drought stress regulation of aquaporins. Plant Mol Biol 59:469–484CrossRefPubMedGoogle Scholar
  3. Amodeo G, Dorr R, Vallejo A, Sutka M, Parisi M (1999) Radial and axial water transport in the sugar beet storage root. J Exp Bot 50:509–516CrossRefGoogle Scholar
  4. Aroca R, Amodeo G, Fernandez-Illescas S, Herman EM, Chaumont F, Chrispeels MJ (2005) The role of aquaporins and membrane damage in chilling and hydrogen peroxide induced changes in the hydraulic conductance of maize roots. Plant Physiol 137:341–353CrossRefPubMedGoogle Scholar
  5. Azaizeh H, Gunse B, Steudle E (1992) Effects of NaCl and CaCl2 on water transport across root-cells of maize (Zea-mays L.) seedlings. Plant Physiol 99:886–894CrossRefPubMedGoogle Scholar
  6. Barrieu F, Chaumont F, Chrispeels MJ (1998) High expression of the tonoplast aquaporin ZmTIP1 in epidermal and conducting tissues of maize. Plant Physiol 117:1153–1163CrossRefPubMedGoogle Scholar
  7. Boursiac Y, Chen S, Luu DT, Sorieul M, van den Dries N, Maurel C (2005) Early effects of salinity on water transport in Arabidopsis roots. Molecular and cellular features of aquaporin expression. Plant Physiol 139:790–805CrossRefPubMedGoogle Scholar
  8. Boyer J (1985) Water transport. Ann Rev Plant Physiol 36:473–516Google Scholar
  9. Bret-Harte MS, Silk WK (1994) Nonvascular, symplasmic diffusion of sucrose cannot satisfy the carbon demands of growth in the primary root tip of Zea mays L. Plant Physiol 105:19–33PubMedGoogle Scholar
  10. Brown D (2003) The ins and outs of aquaporin-2 trafficking. Am J Physiol Renal Physiol 284:F893–F901PubMedGoogle Scholar
  11. Brundrett MC, Enstone DE, Peterson CA (1988) A berberine-aniline blue fluorescent staining procedure for suberin, lignin, and callose in plant-tissue. Protoplasma 146:133–142CrossRefGoogle Scholar
  12. Carvajal M, Cerda A, Martinez V (2000) Does calcium ameliorate the negative effect of NaCl on melon root water transport by regulating aquaporin activity? New Phytol 145:439–447CrossRefGoogle Scholar
  13. Carvajal M, Cooke DT, Clarkson DT (1996) Responses of wheat plants to nutrient deprivation may involve the regulation of water-channel function. Planta 199:372–381CrossRefGoogle Scholar
  14. Carvajal M, Martinez V, Alcaraz CF (1999) Physiological function of water channels as affected by salinity in roots of paprika pepper. Physiol Plant 105:95–101CrossRefGoogle Scholar
  15. Chaumont F, Barrieu F, Jung R, Chrispeels MJ (2000) Plasma membrane intrinsic proteins from maize cluster in two sequence subgroups with differential aquaporin activity. Plant Physiol 122:1025–1034CrossRefPubMedGoogle Scholar
  16. Chaumont F, Barrieu F, Wojcik E, Chrispeels MJ, Jung R (2001) Aquaporins constitute a large and highly divergent protein family in maize. Plant Physiol 125:1206–1215CrossRefPubMedGoogle Scholar
  17. Chaumont F, Moshelion M, Daniels MJ (2005) Regulation of plant aquaporin activity. Biol Cell 97:749–764CrossRefPubMedGoogle Scholar
  18. de Dorlodot S, Bertin P, Baret PV, Draye X (2005) Scaling up quantitative phenotyping of root system architecture using a combination of aeroponics and image analysis. Aspect Appl Biol 73:41–54Google Scholar
  19. Enstone DE, Peterson CA (2005) Suberin lamella development in maize seedling roots grown in aerated and stagnant conditions. Plant Cell Environ 28:444–455CrossRefGoogle Scholar
  20. Fetter K, Van Wilder V, Moshelion M, Chaumont F (2004) Interactions between plasma membrane aquaporins modulate their water channel activity. Plant Cell 16:215–228CrossRefPubMedGoogle Scholar
  21. Frensch J, Hsiao TC, Steudle E (1996) Water and solute transport along developing maize roots. Planta 198:348–355CrossRefGoogle Scholar
  22. Gaspar M, Bousser A, Sissoeff I, Roche O, Hoarau J, Mahe A (2003) Cloning and characterization of ZmPIP1-5b, an aquaporin transporting water and urea. Plant Sci 165:21–31CrossRefGoogle Scholar
  23. Guan KL, Dixon JE (1991) Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem 192:262–267CrossRefPubMedGoogle Scholar
  24. Hose E, Clarkson DT, Steudle E, Schreiber L, Hartung W (2001) The exodermis: a variable apoplastic barrier. J Exp Bot 52:2245–2264CrossRefPubMedGoogle Scholar
  25. Hukin D, Doering-Saad C, Thomas CR, Pritchard J (2002) Sensitivity of cell hydraulic conductivity to mercury is coincident with symplasmic isolation and expression of Plasmalemma aquaporin genes in growing maize roots. Planta 215:1047–1056CrossRefPubMedGoogle Scholar
  26. Ishikawa H, Evans ML (1995) Specialized zones of development in roots. Plant Physiol 109:725–727PubMedGoogle Scholar
  27. Jang JY, Kim DG, Kim YO, Kim JS, Kang H (2004) An expression analysis of a gene family encoding plasma membrane aquaporins in response to abiotic stresses in Arabidopsis thaliana. Plant Mol Biol 54:713–725CrossRefPubMedGoogle Scholar
  28. Javot H, Maurel C (2002) The role of aquaporins in root water uptake. Ann Bot (Lond) 90:301–313CrossRefGoogle Scholar
  29. Johansson I, Karlsson M, Johanson U, Larsson C, Kjellbom P (2000) The role of aquaporins in cellular and whole plant water balance. Biochim Biophys Acta 1465:324–342CrossRefPubMedGoogle Scholar
  30. Karahara I, Ikeda A, Kondo T, Uetake Y (2004) Development of the Casparian strip in primary roots of maize under salt stress. Planta 219:41–47CrossRefPubMedGoogle Scholar
  31. Lopez M, Bousser AS, Sissoeff I, Gaspar M, Lachaise B, Hoarau J, Mahe A (2003) Diurnal regulation of water transport and aquaporin gene expression in maize roots: contribution of PIP2 proteins. Plant Cell Physiol 44:1384–1395CrossRefPubMedGoogle Scholar
  32. Luu DT, Maurel C (2005) Aquaporins in a challenging environment: molecular gears for adjusting plant water status. Plant Cell Environ 28:85–96CrossRefGoogle Scholar
  33. Maggio A, Joly RJ (1995) Effects of mercuric chloride on the hydraulic conductivity of tomato root systems (evidence for a channel-mediated water pathway). Plant Physiol 109:331–335PubMedGoogle Scholar
  34. Marino JH, Cook P, Miller KS (2003) Accurate and statistically verified quantification of relative mRNA abundances using SYBR Green I and real-time RT-PCR. J Immunol Methods 283:291–306CrossRefPubMedGoogle Scholar
  35. Martre P, Morillon R, Barrieu F, North GB, Nobel PS, Chrispeels MJ (2002) Plasma membrane aquaporins play a significant role during recovery from water deficit. Plant Physiol 130:2101–2110CrossRefPubMedGoogle Scholar
  36. Martre P, North GB, Nobel PS (2001) Hydraulic conductance and mercury-sensitive water transport for roots of Opuntia acanthocarpa in relation to soil drying and rewetting. Plant Physiol 126:352–362CrossRefPubMedGoogle Scholar
  37. Maurel C, Chrispeels MJ (2001) Aquaporins. A molecular entry into plant water relations. Plant Physiol 125:135–138CrossRefPubMedGoogle Scholar
  38. Moore R, Smith HS (1990) Morphometric analysis of epidermal differentiation in primary roots of Zea-mays. Am J Bot 77:727–735CrossRefPubMedGoogle Scholar
  39. Morsomme P, de Kerchove d’Exaerde A, De Meester S, Thines D, Goffeau A, Boutry M (1996) Single point mutations in various domains of a plant plasma membrane H(+)-ATPase expressed in Saccharomyces cerevisiae increase H(+)-pumping and permit yeast growth at low pH. Embo J 15:5513–5526PubMedGoogle Scholar
  40. Moshelion M, Moran N, Chaumont F (2004) Dynamic changes in the osmotic water permeability of protoplast plasma membrane. Plant Physiol 135:2301–2317CrossRefPubMedGoogle Scholar
  41. Nuovo GJ (1996) The foundations of successful RT in situ PCR. Front Biosci 1:c4–c15PubMedGoogle Scholar
  42. Oparka KJ, Duckett CM, Prior DAM, Fisher DB (1994) Real-time imaging of phloem unloading in the root-tip of arabidopsis. Plant J 6:759–766CrossRefGoogle Scholar
  43. Otto B, Kaldenhoff R (2000) Cell-specific expression of the mercury-insensitive plasma-membrane aquaporin NtAQP1 from Nicotiana tabacum. Planta 211:167–172CrossRefPubMedGoogle Scholar
  44. Patrick JW, Offler CE (1996) Post-sieve element transport of photoassimilates in sink regions. J Exp Bot 47:1165–1177Google Scholar
  45. Perumalla CJ, Peterson CA (1986) Deposition of Casparian bands and suberin lamellae in the exodermis and endodermis of young corn and onion roots. Can J Bot 64:1873–1878CrossRefGoogle Scholar
  46. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45CrossRefPubMedGoogle Scholar
  47. Ramahaleo T, Morillon R, Alexandre J, Lassalles JP (1999) Osmotic water permeability of isolated protoplasts. Modifications during development. Plant Physiol 119:885–896CrossRefPubMedGoogle Scholar
  48. Raven PH, Evert RF, Eichhorn SE (1992) The movement of water and solutes in plants. In: Mastalski SAE (ed) Biology of plants. Worth Publishers, New YorkGoogle Scholar
  49. Schaffner AR (1998) Aquaporin function, structure, and expression: are there more surprises to surface in water relations? Planta 204:131–139CrossRefPubMedGoogle Scholar
  50. Schraut D, Ullrich CI, Hartung W (2004) Lateral ABA transport in maize roots (Zea mays): visualization by immunolocalization. J Exp Bot 55:1635–1641CrossRefPubMedGoogle Scholar
  51. Steudle E (2000) Water uptake by plant roots: an integration of views. Plant Soil 226:45–56CrossRefGoogle Scholar
  52. Steudle E (2001) The cohesion-tension mechanism and the acquisition of water by plant roots. Annu Rev Plant Physiol Plant Mol Biol 52:847–875CrossRefPubMedGoogle Scholar
  53. Suga S, Imagawa S, Maeshima M (2001) Specificity of the accumulation of mRNAs and proteins of the plasma membrane and tonoplast aquaporins in radish organs. Planta 212:294–304CrossRefPubMedGoogle Scholar
  54. Suga S, Komatsu S, Maeshima M (2002) Aquaporin isoforms responsive to salt and water stresses and phytohormones in radish seedlings. Plant Cell Physiol 43:1229–1237CrossRefPubMedGoogle Scholar
  55. Tazawa M, Ohkuma E, Shibasaka M, Nakashima S (1997) Mercurial-sensitive water transport in barley roots. J Plant Res 110:435–442CrossRefGoogle Scholar
  56. Tyerman SD, Bohnert HJ, Maurel C, Steudle E, Smith JAC (1999) Plant aquaporins: their molecular biology, biophysics and significance for plant water relations. J Exp Bot 50:1055–1071CrossRefGoogle Scholar
  57. Tyerman SD, Niemietz CM, Bramley H (2002) Plant aquaporins: multifunctional water and solute channels with expanding roles. Plant Cell Environ 25:173–194CrossRefPubMedGoogle Scholar
  58. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:RESEARCH0034Google Scholar
  59. Zhu C, Schraut D, Hartung W, Schaffner AR (2005) Differential responses of maize MIP genes to salt stress and ABA. J Exp Bot 56:2971–2981CrossRefPubMedGoogle Scholar
  60. Zhu GL, Steudle E (1991) Water transport across maize roots—simultaneous measurement of flows at the cell and root level by double pressure probe technique. Plant Physiol 95:305–315CrossRefPubMedGoogle Scholar
  61. Zimmermann HM, Hartmann K, Schreiber L, Steudle E (2000) Chemical composition of apoplastic transport barriers in relation to radial hydraulic conductivity of corn roots (Zea mays L.). Planta 210:302–311CrossRefPubMedGoogle Scholar
  62. Zimmermann HM, Steudle E (1998) Apoplastic transport across young maize roots: effect of the exodermis. Planta 206:7–19CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Charles Hachez
    • 1
  • Menachem Moshelion
    • 1
    • 2
  • Enric Zelazny
    • 1
  • Damien Cavez
    • 1
  • François Chaumont
    • 1
  1. 1.Unité de Biochimie physiologiqueInstitut des Sciences de la Vie, Université catholique de LouvainLouvain-la-NeuveBelgium
  2. 2.The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agriculture, Food & Environment Quality ScienceThe Hebrew UniversityRehovotIsrael

Personalised recommendations