Salinity-mediated transcriptional and post-translational regulation of the Arabidopsis aquaporin PIP2;7

  • 1156 Accesses

  • 13 Citations


Key message

Salt stress triggers a simultaneous transcriptional repression and aquaporin internalization to modify root cell water conductivity.


Plasma membrane intrinsic proteins (PIPs) are involved in the adjustment of plant water balance in response to changing environmental conditions. In this study, Arabidopsis wild-type (Col-0) and transgenic lines overexpressing PIP2;7 were used to investigate and compare their response to salt stress. Hydraulic conductivity measurements using a high-pressure flowmeter (HPFM) revealed that overexpression of PIP2;7 induced a sixfold increase in root hydraulic conductivity of four week-old Arabidopsis thaliana plants compared to WT. Exposure to a high salt stress (150 mM NaCl) triggered a rapid repression of overall aquaporin activity in both genotypes. Response to salt stress was also investigated in 8 day-old seedlings. Exposure to salt led to a repression of PIP2;7 promoter activity and a significant decrease in PIP2;7 mRNA abundance within 2 h. Concomitantly, a rapid internalization of fluorescently-tagged PIP2;7 proteins was observed but removal from the cell membrane was not accompanied by further degradation of the protein within 4 h of exposure to salinity stress. These data suggest that PIP transcriptional repression and channel internalization act in concert during salt stress conditions to modulate aquaporin activity, thereby significantly altering the plant hydraulic parameters in the short term.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 199

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. Aharon R, Shahak Y, Wininger S, Bendov R, Kapulnik Y, Galili G (2003) Overexpression of a plasma membrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but to under drought or salt stress. Plant Cell 15:439–447

  2. Alexandersson E, Fraysse L, Sjövall-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 59:469–484

  3. Alexandersson E, Danielson JA, Rade J, Moparthi VK, Fontes M, Kjellbom P, Johanson U (2010) Transcriptional regulation of aquaporins in accessions of Arabidopsis in response to drought stress. Plant J 61:650–660

  4. Aroca R, Ferrante A, Vernieri P, Chrispeels MJ (2006) Drought, abscisic acid and transpiration rate effects on the regulation of PIP Aquaporin Gene expression and abundance in Phaseolus vulgaris plants. Ann Bot (Lond) 98:1301–1310

  5. Azaizeh H, Steudle E (1991) Effects of salinity on water transport of excised maize (Zea-Mays L) roots. Plant Physiol 97:1136–1145

  6. Bogeat-Triboulot M-B, Brosche M, Renaut J, Jouve L, Le Thiec D, Fayyaz P, Vinocur B, Witters E, Laukens K, Teichmann T, Altman A, Hausman J-F, Polle A, Kangasjarvi J, Dreyer E (2007) Gradual soil water depletion results in reversible changes of gene expression, protein profiles, ecophysiology, and growth performance in Populus euphratica, a poplar growing in arid regions. Plant Physiol 143:876–892

  7. Bots M, Vergeldt F, Wolters-Arts M, Weterings K, van As H, Mariani C (2005) Aquaporins of the PIP2 class are required for efficient anther dehiscence in tobacco. Plant Physiol 137:1049–1056

  8. 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 139:790–805

  9. Boursiac Y, Boudet J, Postaire O, Luu DT, Tournaire-Roux C, Maurel C (2008) Stimulus-induced downregulation of root water transport involves reactive oxygen species-activated cell signalling and plasma membrane intrinsic protein internalization. Plant J 56:207–218

  10. Brugnoli E, Lauteri M (1991) Effects of salinity on stomatal conductance, photosynthetic capacity, and carbon ısotope discrimination of salt-resistant (Gossypium hirsutum L.) and saltsensitive (Phaseolus vulgaris L.) C3 non-halophytes. Plant Physiol 95:628–635

  11. Caldeira CF, Bosio M, Parent B, Jeanguenin L, Chaumont F, Tardieu F (2014a) A hydraulic model is compatible with rapid changes in leaf elongation under fluctuating evaporative demand and soil water status. Plant Physiol 164:1718–1730

  12. Caldeira CF, Jeanguenin L, Chaumont F, Tardieu F (2014b) Circadian rhythms of hydraulic conductance and growth are enhanced by drought and improve plant performance. Nat Commun 5:5365

  13. Calvo-Polanco M, Sanchez-Romera B, Aroca R (2014) Mild salt stress conditions induce different responses in root hydraulic conductivity of phaseolus vulgaris over-time. PLoS One 9:e90631

  14. Chang W, Liu X, Zhu J, Fan W, Zhang Z (2016) An aquaporin gene from halophyte Sesuvium portulacastrum, SpAQP1, increases salt tolerance in transgenic tobacco. Plant Cell Rep 35:385–395

  15. Chaumont F, Tyerman SD (2014) Aquaporins: highly regulated channels controlling plant water relations. Plant Physiol 164:1600–1618

  16. Conn SJ, Hocking B, Dayod M, Xu B, Athman A, Henderson S, Aukett L, Conn V, Shearer MK, Fuentes S, Tyerman SD, Gilliham M (2013) Protocol: optimising hydroponic growth systems for nutritional and physiological analysis of Arabidopsis thaliana and other plants. Plant Methods 9:4

  17. Di Pietro M, Vialaret J, Li GW, Hem S, Prado K, Rossignol M, Maurel C, Santoni V (2013) Coordinated post-translational responses of aquaporins to abiotic and nutritional stimuli in Arabidopsis roots. Mol Cell Proteomics 12:3886–3897

  18. Ding X, Iwasaki I, Kitagawa Y (2004) Overexpression of a lily PIP1 gene in tobacco increased the osmotic water permeability of leaf cells. Plant Cell Environ 27:177–186

  19. Galmes J, Pou A, Alsina M, Tomas M, Medrano H, Flexas J (2007) Aquaporin expression in response to different water stress intensities and recovery in Richter-110 (Vitis sp.): relationship with ecophysiological status. Planta 226:671–681

  20. Gao Z, He X, Zhao B, Zhou C, Liang Y, Ge R (2010) Over-expressing a putative aquaporin gene from wheat, TaNIP, enhances salt tolerance in transgenic Arabidopsis. Plant Cell Physiol 51:767–775

  21. Guo L, Wang ZY, Lin H, Cui WE, Chen J, Liu M (2006) Expression and functional analysis of the rice plasma-membrane intrinsic protein gene family. Cell Res 16:277–286

  22. Hachez C, Moshelion M, Zelazny E, Cavez D, Chaumont F (2006) Localization and quantification of plasma membrane aquaporin expression in maize primary root: a clue to understanding their role as cellular plumbers. Plant Mol Biol 62:305–323

  23. Hachez C, Veselov D, Ye Q, Reinhardt H, Knipfer T, Fricke W, Chaumont F (2012) Short-term control of maize cell and root water permeability through plasma membrane aquaporin isoforms. Plant Cell Environ 35:185–198

  24. Hachez C, Besserer A, Chevalier AS, Chaumont F (2013) Insights into plant plasma membrane aquaporin trafficking. Trends Plant Sci 18:344–352

  25. Hachez C, Laloux T, Reinhardt H, Cavez D, Degand H, Grefen C, De Rycke R, Inze D, Blatt MR, Russinova E, Chaumont F (2014a) Arabidopsis SNAREs SYP61 and SYP121 coordinate the trafficking of plasma membrane aquaporin PIP2;7 to modulate the cell membrane water permeability. Plant Cell 26:3132–3147

  26. Hachez C, Veljanovski V, Reinhardt H, Guillaumot D, Vanhee C, Chaumont F, Batoko H (2014b) The Arabidopsis abiotic stress-induced TSPO-related protein reduces cell-surface expression of the aquaporin PIP2;7 through protein–protein interactions and autophagic degradation. Plant Cell 26:4974–4990

  27. Hanba YT, Shibasaka M, Hayashi Y, Hayakawa T, Kasamo K, Terashima I, Katsuhara M (2004) Overexpression of the Barley aquaporin in HvPIP2;1 increases internal CO(2) conductance and CO(2) assimilation in the leaves of transgenic rice plants. Plant Cell Physiol 45:521–529

  28. Hilal M, Zenoff AM, Ponessa G, Moreno H, Massa EM (1998) Saline stress alters the temporal patterns of xylem differentiation and alternative oxidase expression in developing soybean roots. Plant Physiol 117:695–701

  29. Horie T, Kaneko T, Sugimoto G, Sasano S, Panda SK, Shibasaka M, Katsuhara M (2011) Mechanisms of water transport mediated by PIP aquaporins and their regulation via phosphorylation events under salinity stress in barley roots. Plant Cell Physiol 52:663–675

  30. 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 54:713–725

  31. Javot H, Lauvergeat V, Santoni V, Martin-Laurent F, Guclu J, Vinh J, Heyes J, Franck KI, Schaffner AR, Bouchez D, Maurel C (2003) Role of a single Aquaporin isoform in root water uptake. Plant Cell 15:509–522

  32. Johanson U, Karlsson M, Johansson I, Gustavsson S, Sjovall S, Fraysse L, Weig AR, Kjellbom P (2001) The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants. Plant Physiol 126:1358–1369

  33. Johansson I, Karlsson M, Shukla VK, Chrispeels MJ, Larsson C, Kjellbom P (1998) Water transport activity of the plasma membrane aquaporin PM28A is regulated by phosphorylation. Plant Cell 10:451–459

  34. Kilian J, Whitehead D, Horak J, Wanke D, Weinl S, Batistic O, D’Angelo C, Bornberg-Bauer E, Kudla J, Harter K (2007) The AtGenExpress global stress expression data set: protocols, evaluation and model data analysis of UV-B light, drought and cold stress responses. Plant J 50:347–363

  35. Lee SH, Zwiazek JJ (2015) Regulation of aquaporin-mediated water transport in Arabidopsis roots exposed to NaCl. Plant Cell Physiol 56:750–758

  36. Li X, Wang X, Yang Y, Li R, He Q, Fang X, Luu DT, Maurel C, Lin J (2011) Single-molecule analysis of PIP2;1 dynamics and partitioning reveals multiple modes of Arabidopsis plasma membrane aquaporin regulation. Plant Cell 23:3780–3797

  37. Lian HL, Yu X, Ye Q, Ding X, Kitagawa Y, Kwak SS, Su WA, Tang ZC (2004) The role of aquaporin RWC3 in drought avoidance in rice. Plant Cell Physiol 45:481–489

  38. 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 44:1384–1395

  39. Luu DT, Martiniere A, Sorieul M, Runions J, Maurel C (2012) Fluorescence recovery after photobleaching reveals high cycling dynamics of plasma membrane aquaporins in Arabidopsis roots under salt stress. Plant J 69:894–905

  40. Martinez-Ballesta MC, Aparicio F, Pallas V, Martinez V, Carvajal M (2003) Influence of saline stress on root hydraulic conductance and PIP expression in Arabidopsis. J Plant Physiol 160:689–697

  41. Martiniere A, Li X, Runions J, Lin J, Maurel C, Luu DT (2012) Salt stress triggers enhanced cycling of Arabidopsis root plasma-membrane aquaporins. Plant Signal Behav 7:524–532

  42. Marulanda A, Azcon R, Chaumont F, Ruiz-Lozano JM, Aroca R (2010) Regulation of plasma membrane aquaporins by inoculation with a Bacillus megaterium strain in maize (Zea mays L.) plants under unstressed and salt-stressed conditions. Planta 232:533–543

  43. Michael TP, Breton G, Hazen SP, Priest H, Mockler TC, Kay SA, Chory J (2008) A morning-specific phytohormone gene expression program underlying rhythmic plant growth. PLoS Biol 6:e225

  44. Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681

  45. Muries B, Faize M, Carvajal M, Martinez-Ballesta M del C (2011) Identification and differential induction of the expression of aquaporins by salinity in broccoli plants. Mol Biosyst 7:1322–1335

  46. Nühse TS, Stensballe A, Jensen ON, Peck SC (2004) Phosphoproteomics of the Arabidopsis plasma membrane and a new phosphorylation site database. Plant Cell 16:2394–2405

  47. Pih KT, Kabilan V, Lim JH, Kang SG, Piao HL, Jin JB, Hwang I (1999) Characterization of two new channel protein genes in Arabidopsis. Mol Cells 9:84–90

  48. Postaire O, Tournaire-Roux C, Grondin A, Boursiac Y, Morillon R, Schaffner AR, Maurel C (2010) A PIP1 aquaporin contributes to hydrostatic pressure-induced water transport in both the root and rosette of Arabidopsis. Plant Physiol 152:1418–1430

  49. Prado K, Boursiac Y, Tournaire-Roux C, Monneuse JM, Postaire O, Da Ines O, Schaffner AR, Hem S, Santoni V, Maurel C (2013) Regulation of Arabidopsis leaf hydraulics involves light-dependent phosphorylation of aquaporins in veins. Plant Cell 25:1029–1039

  50. Prak S, Hem S, Boudet J, Viennois G, Sommerer N, Rossignol M, Maurel C, Santoni V (2008) Multiple phosphorylations in the C-terminal tail of plant plasma membrane aquaporins: role in subcellular trafficking of AtPIP2;1 in response to salt stress. MolCell Prot 7:1019–1030

  51. Qian ZJ, Song JJ, Chaumont F, Ye Q (2015) Differential responses of plasma membrane aquaporins in mediating water transport of cucumber seedlings under osmotic and salt stresses. Plant Cell Environ 38:461–473

  52. Sade N, Vinocur BJ, Diber A, Shatil A, Ronen G, Nissan H, Wallach R, Karchi H, Moshelion M (2009) Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion? New Phytol 181:651–661

  53. Sakurai J, Ishikawa F, Yamaguchi T, Uemura M, Maeshima M (2005) Identification of 33 rice aquaporin genes and analysis of their expression and function. Plant Cell Physiol 46:1568–1577

  54. Sakurai-Ishikawa J, Murai-Hatano M, Hayashi H, Ahamed A, Fukushi K, Matsumoto T, Kitagawa Y (2011) Transpiration from shoots triggers diurnal changes in root aquaporin expression. Plant Cell Environ 34:1150–1163

  55. Siefritz F, Tyree MT, Lovisolo C, Schubert A, Kaldenhoff R (2002) PIP1 plasma membrane aquaporins in tobacco: from cellular effects to function in plants. Plant Cell 14:869–876

  56. Sutka M, Li G, Boudet J, Boursiac Y, Doumas P, Maurel C (2011) Natural variation of root hydraulics in Arabidopsis grown in normal and salt stress conditions. Plant Physiol 155:1264–1276

  57. Takase T, Ishikawa H, Murakami H, Kikuchi J, Sato-Nara K, Suzuki H (2011) The circadian clock modulates water dynamics and aquaporin expression in Arabidopsis roots. Plant Cell Physiol 52:373–383

  58. Taylaran RD, Adachi S, Ookawa T, Usuda H, Hirasawa T (2011) Hydraulic conductance as well as nitrogen accumulation plays a role in the higher rate of leaf photosynthesis of the most productive variety of rice in Japan. J Exp Bot 62:4067–4077

  59. Tornroth-Horsefield S, Wang Y, Hedfalk K, Johanson U, Karlsson M, Tajkhorshid E, Neutze R, Kjellbom P (2006) Structural mechanism of plant aquaporin gating. Nature 439:688–694

  60. Tyree MT, Patiño S, Benink J, Alexander J (1995) Dynamic measurements of root hydraulic conductance using a high pressure flowmeter in the laboratory and field. J Exp Bot 46:83–94

  61. Uehlein N, Lovisolo C, Siefritz F, Kaldenhoff R (2003) The tobacco aquaporin NtAQP1 is a membrane CO2 pore with physiological functions. Nature 425:734–737

  62. Van Wilder V, Miecielica U, Degand H, Derua R, Waelkens E, Chaumont F (2008) Maize plasma membrane aquaporins belonging to the PIP1 and PIP2 subgroups are in vivo phosphorylated. Plant Cell Physiol 49:1364–1377

  63. Vandeleur RK, Mayo G, Shelden MC, Gilliham M, Kaise BN, Tyerman S (2009) The role of plasma membrane intrinsic protein aquaporins in water transport through roots: diurnal and drought stress responses reveal different strategies between isohydric and anisohydric cultivars of grapevine. Plant Physiol 149:445–460

  64. Vialaret J, Di Pietro M, Hem S, Maurel C, Rossignol M, Santoni V (2014) Phosphorylation dynamics of membrane proteins from Arabidopsis roots submitted to salt stress. Proteomics 14:1058–1070

  65. Winter D, Vinegar B, Nahal H, Ammar R, Wilson GV, Provart NJ (2007) An electronic florescent pictograph browser for exploring and analyzing large-scale biological data sets. PLoS One 2:e718

  66. Yamamoto N, Takano T, Tanaka K, Ishige T, Terashima S, Endo C, Kurusu T, Yajima S, Yano K, Tada Y.(2015) Comprehensive analysis of transcriptome response to salinity stress in the halophytic turf grass Sporobolus virginicus. Front Plant Sci 2015 6:241.

  67. Zhu C, Schraut D, Hartung W, Schäffner AR (2005) Differential responses of maize MIP genes to salt stress and ABA. J Exp Bot 56(2):971–2981

Download references


We thank Ewelina Grzeskiewicz for her help in this work and Frank Van Breusegem (PSB, VIB, Ghent) for lending the Licor infrared gas-exchange system. Confocal microscopy was carried out at the UCL imaging platform IMABIOL. This work was supported by grants from the Belgian National Fund for Scientific Research (FNRS), the Interuniversity Attraction Poles Programme-Belgian Science Policy (IAP7/29), the Belgian French community ARC11/16–036 project and the Bauchau Award. A. P. was supported by an UCL Incoming post-doctoral Fellowship co-funded by the Marie Curie Actions of the European Commission. C.H. was a FNRS postdoctoral researcher. T.M. was a research fellow at the Fonds de Formation à la Recherche dans l’Industrie et l’Agriculture.

Author contributions

Alicia Pou: Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting and revising the article. Linda Jeanguenin: Acquisition of data, Analysis and interpretation of data, Drafting and revising the article. Thomas Milhiet: Acquisition of data, Analysis and interpretation of data. Henri Batoko: Drafting and revising the article. François Chaumont: Conception and design, Analysis and interpretation of data, Drafting and revising the article. Charles Hachez: Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting and revising the article.

Author information

Correspondence to François Chaumont or Charles Hachez.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Lack of diurnal variation in transcript abundance of GUS gene under control conditions. RNA was extracted from an homozygous T3 line expressing the PIP2;7:nlsGFP-GUS construct (Hachez et al., 2014a) at different time points. The GUS transcript abundance showed no statistically significant (P>0.05) diurnal variation. The three genes used for normalization were the following ones: Elongation Factor (EF1α), SAND family protein (SFP) and protein phosphatase 2A3 (PP2A3). Data points represent means and standard deviation of four replicates from two independent biological replicates (EPS 755 KB)

Transcript abundance of control genes under salt stress. Ct values of gene-specific primers used for data normalization in RT-qPCR. The transcript abundance remained unaltered during salt stress. Genes: Elongation Factor (EF1α), SAND family protein (SFP) and protein phosphatase 2A3 (PP2A3) (EPS 955 KB)

Stomatal conductance, photosynthetic rate and transpiration in 30 day-old plants grown in soil. (A) Response of stomatal conductance for CO2 (g s ) after 4 h of salt stress in WT (Col-0) and 35S:Venus-PIP2;7 plants. Data points represent means and standard errors of at least six replicates. (B) Response of photosynthetic CO2 assimilation (A N ) after 4 h of salt stress in Col-0 (WT) and 35S:Venus-PIP2;7 plants. (C) Response of leaf transpiration (E) after 4 h of salt stress in Col-0 (WT) and Venus-PIP2;7 plants. Data points represent means and standard errors of at least six replicates. Different letters represent significant differences within treatments for both genotypes by two-way Anova with Tukey’s test (P < 0.05). (EPS 1063 KB)

Supplementary material 4 (DOCX 12 KB)

Time-lapse analysis of the Venus-PIP2;7 fluorescence in root PM. Arabidopsis root cells expressing the PIP2;7:Venus-PIP2;7 construct were monitored by confocal microscopy and images were taken during an interval of 76 s. Note the presence of PM invaginations in cortical root cells (AVI 447 KB)

Forty seven min time-lapse analysis of the Venus-PIP2;7 fluorescence in root PM. Arabidopsis root cells expressing the PIP2;7:Venus-PIP2;7 construct were monitored by confocal microscopy and images were taken every 6 min during an interval of 48 min. Recording started at the onset of the stress. Note 1/the decrease in fluorescence of Venus-PIP2;7 in the PM 2/ the concomitant cell plasmolysis and 3/ the presence of PM invaginations in cortical root cells in response to this salt treatment (AVI 1064 KB)

Forty seven min time-lapse analysis of the Venus-PIP2;7 fluorescence in root PM. Arabidopsis root cells expressing the PIP2;7:Venus-PIP2;7 construct were monitored by confocal microscopy and images were taken every 6 min during an interval of 48 min. Recording started at the onset of the stress. Note 1/the decrease in fluorescence of Venus-PIP2;7 in the PM 2/ the concomitant cell plasmolysis and 3/ the presence of PM invaginations in cortical root cells in response to this salt treatment (AVI 1064 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Pou, A., Jeanguenin, L., Milhiet, T. et al. Salinity-mediated transcriptional and post-translational regulation of the Arabidopsis aquaporin PIP2;7. Plant Mol Biol 92, 731–744 (2016) doi:10.1007/s11103-016-0542-z

Download citation


  • Aquaporin
  • Plasma membrane
  • Root hydraulic conductivity
  • Salt stress
  • Water relation