The ability to respond quickly to salt stress can determine the tolerance level of a species. Here, we test how rapidly the roots of Calotropis procera react to high salinity conditions. In the first 24 h after saline exposure, the plants reduced stomatal conductance, increased CO2 assimilation, and water use efficiency. Thus, the root tissue showed an immediate increase in soluble sugars, free amino acid, and soluble protein contents. Twelve aquaporins showed differential gene expression in the roots of C. procera under salinity. Transcriptional upregulation was observed only after 2 h, with greater induction of CpTIP1.4 (fourfold). Transcriptional downregulation, in turn, occurred mainly after 8 h, with the largest associated with CpPIP1.2 (fourfold). C. procera plants responded quickly to high saline levels. Our results showed a strong stomatal control associated with high free amino acid and soluble sugar contents, regulated aquaporin expression in roots, and supported the high performance of the root system of C. procera under salinity. Moreover, this species was able to maintain a lower Na+/K+ ratio in the leaves compared to that of the roots of stressed plants. The first response of the root system, after immediate contact with saline solution, present an interesting scenario to discuss.
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An J, Hu Z, Che B, Chen H, Yu B, Cai W (2017) Heterologous expression of Panax ginseng PgTIP1 confers enhanced salt tolerance of soybean cotyledon hairy roots, composite, and whole plants. Front Plant Sci 8:1232. https://doi.org/10.3389/fpls.2017.01232
Aroca R, Amoedo G, Fernandéz-Ilescas S, Herman EM, Chaumont F, Chrispeels M (2005) The role of aquaporins and membrane damage in chilling and hydrogen peroxide induced changes in the hydraulic conductance of maize roots. Plant Physiol 137:341–353. https://doi.org/10.1104/pp.104.051045
Barrs HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci 15:413–428. https://doi.org/10.1071/bi9620413
Bellati J, Alleva K, Soto G, Vitali V, Josefkowicz C, Amoedo G (2010) Intracellular pH sensing is altered by plasma membrane PIP aquaporin co-expression. Plant Mol Biol 74:105–118. https://doi.org/10.1007/BF00018060
Bezerra-Neto P, Araújo FC, Ferreira-Neto JRC, Silva MD, Pandolfi V, Aburjaile FF, Sakamoto T, Silva RLO, Kido EA, Amorim LLB, Ortega JM, Benko-Iseppon AM (2019) Plant Aquaporins: diversity, evolution and biotechnological applications. Curr Protein Pept Sci 20:368–395. https://doi.org/10.1007/s11103-010-9658-8
Biassoni R (2014) Quantitative Real-Time PCR: Methods and Protocols. In: Biassoni R, Raso A (eds) Methods in Molecular Biology. Springer, New York, p 1160
Blum A (2017) Osmotic adjustment is a prime drought stress adaptive engine in support of plant production. Plant Cell Environ 40:4–10. https://doi.org/10.1111/pce.12800
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Campbell GS, Norman JM (1998) An introduction to environmental biophysics. Springer, New York
Coêlho MRV, Rivas R, Ferreira-Neto JCR, Pandolfi V, Bezerra-Neto JP, Benko-Iseppon AM, Santos MG (2019) Reference genes selection for Calotropis procera under different salt stress conditions. PLoS ONE 14:e0215729. https://doi.org/10.1371/journal.pone.0215729
Dreyer I, Uozumi N (2011) Potassium channels in plant cells. FEBS J 278:4293–4303. https://doi.org/10.1111/j.1742-4658.2011.08371.x
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356
Flexas J, Diaz-Espejo A, Conesa MA, Coopman RE, Douthe C, Gago J, Gallé A, Galmés J, Medrano H, Ribas-Carbo M, Tomàs M, Niinemets U (2016) Mesophyll conductance to CO2 and Rubisco as targets for improving intrinsic water use efficiency in C3 plants. Plant Cell Environ 39:965–982. https://doi.org/10.1111/pce.12622
Han W, Jia J, Hu Y, Liu J, Guo J, Shi Yu, Huo H, Gong H (2020) Maintenance of root water uptake contributes to salt-tolerance of a wild tomato species under salt stress. Arch Agron Soil Sci. https://doi.org/10.1080/03650340.2020.1720911
Hassan LM, Galal TM, Farahat EA, El-Midany MM (2015) The biology of Calotropis procera (Aiton) W.T. Trees 29:311–320. https://doi.org/10.1007/s00468-015-1158-7
Johanson U, Karisson 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. https://doi.org/10.1104/pp.126.4.1358
Kapilan R, Vaziri M, Zwiazek JJ (2018) Regulation of aquaporins in plants under stress. Biol Res. https://doi.org/10.1186/s40659-018-0152-0
Li G, Santoni V, Maurel C (2014) Plant aquaporins: Roles in plant physiology. Biochimica et Biophysica Acta (BBA) - General Subjects. Aquaporins 1840:1574–1582. https://doi.org/10.1016/j.bbagen.2013.11.004
Lichtenthaler H, Buschmann C (2001) Chlorophylls and carotenoids: measurement and characterization by UV‐VIS spectroscopy. In: Current protocols in food analytical chemistry, pp 1–8. Doi: https://doi.org/10.1002/0471142913.faf0403s01
Liu C, Li C, Liang D, Wei Z, Zhou S, Wang R, Ma F (2012) Differential expression of ion transporters and aquaporins in leaves may contribute to different salt tolerance in Malus species. Plant Physiol Biochem 58:159–165. https://doi.org/10.1016/j.plaphy.2012.06.019
Mansour MMF, Ali EF (2017) Evaluation of proline functions in saline conditions. Phytochemistry 140:52–68. https://doi.org/10.1016/j.phytochem.2017.04.016
Martínez-Noël GMA, Tognetti JA (2018) Sugar signaling under abiotic stress in plants. In: Ahmad P, Ahanger MA, Singh VP, Tripathi DK, Alam P, Alyemeni MN (eds) Plant metabolites and regulation under environmental stress. Academic Press, New York, pp 397–406
Maurel C, Verdouck L, Luu DT, Santoni V (2008) Plant aquaporins: membrane channels with multiple integrated functions. Ann Rev Plant Biol 59:595–624. https://doi.org/10.1146/annurev.arplant.59.032607.092734
Maurel C, Boursiac Y, Luu DT, Santoni V, Shahzad Z, Verdoucq L (2015) Aquaporins in plants. Physiol Rev 95:1321–1358. https://doi.org/10.1152/physrev.00008.2015
Moore S, Stein WH (1948) Photometric ninhydrin method for use in the chromatography of amino acids. J Biol Chem 176:367–388
Moshelion M, Halperin O, Wallach R, Oren R, Way D (2015) Role of aquaporins in determining transpiration and photosynthesis in water-stressed plants: crop water-use efficiency, growth and yield. Plant Cell Environ 38:1785–1793. https://doi.org/10.1111/pce.12410
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681. https://doi.org/10.1146/annurev.arplant.59.032607.092911
Muries B, Faize M, Carvajal M, Martínez-Ballesta MC (2011) Identification and differential induction of the expression of aquaporins by salinity in broccoli plants. Mol BioSyst 7:1322–1335. https://doi.org/10.1039/c0mb00285b
Mutwakil MZ, Hajrah NH, Atef A, Edris S, Sabir MJ, Al-Ghamdi AK, Sabir MJSM, Nelson C, Makki RM, Ali HM, El-Domyati FM, Al-Hajar ASM, Gloaguen Y, Al-Zahrani HS, Sabir JSM, Jansen RK, Bahieldin A, Hall N (2017) Transcriptomic and metabolic responses of Calotropis procera to salt and drought stress. BMC Plant Biol 17:231. https://doi.org/10.1186/s12870-017-1155-7
Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucl Acids Res 30:e36. https://doi.org/10.1093/nar/30.9.e36
Pou A, Jeanguenin L, Milhiet T, Batoko H, Chaumont F, Hachez C (2016) Salinity-mediated transcriptional and post-translational regulation of the Arabidopsis aquaporin PIP2;7. Plant Mol Biol 92:731–744. https://doi.org/10.1007/s11103-016-0542-z
Rasmussen R (2001) Quantification on the LightCycler. In: Meuer S, Wittwer C, Nakagawara K-I (Eds) Rapid cycle real-time PCR: methods and applications. Springer, Berlin, pp 21–34. Doi: https://doi.org/10.1007/978-3-642-59524-0_3
Rivas R, Barros V, Falcão H, Frosi G, Arruda E, Santos M (2020) Ecophysiological traits of invasive C3 species Calotropis procera to maintain high photosynthetic performance under high VPD and low soil water balance in semi-arid and seacoast zones. Front Plant Sci 11:717. https://doi.org/10.3389/fpls.2020.00717
Rivas R, Frosi G, Ramos DG, Pereira S, Benko-Iseppon AM, Santos MG (2017) Photosynthetic limitation and mechanisms of photoprotection under drought and recovery of Calotropis procera, an evergreen C3 from arid regions. Plant Physiol Biochem 118:589–599. https://doi.org/10.1016/j.plaphy.2017.07.026
Rodríguez-Gamir J, Xue J, Clearwater MJ, Meason DF, Clinton PW, Domec JC (2019) Aquaporin regulation in roots controls plant hydraulic conductance, stomatal conductance, and leaf water potential in Pinus radiata under water stress. Plant Cell Environment 42:717–729. https://doi.org/10.1111/pce.13460
Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana Press, Totowa, pp 365–386
Sami F, Yusuf M, Faizan M, Faraz A, Hayat S (2016) Role of sugars under abiotic stress. Plant Physiol Biochem 109:54–61. https://doi.org/10.1016/j.plaphy.2016.09.005
Secchi F, Zwieniecki MA (2011) Sensing embolism in xylem vessels: the role of sucrose as a trigger for refilling. Plant, Cell Environ 34:514–524
Secchi F, Pagliarani C, Zwieniecki MA (2017) The functional role of xylem parenchyma cells and aquaporins during recovery from severe water stress. Plant Cell Environ 40:858–871. https://doi.org/10.1111/j.1365-3040.2010.02259.x
Shavrukov Y (2013) Salt stress or salt shock: which genes are we studying? J Exp Bot 64:119–127. https://doi.org/10.1093/jxb/ers316
Singh RK, Deshmukh R, Muthamilarasan M, Rani R, Prasad M (2020) Versatile roles of aquaporin in physiological processes and stress tolerance in plants. Plant Physiol Biochem 149:178–189. https://doi.org/10.1016/j.plaphy.2020.02.009
Skorupa-Klaput M, Szczepanek J, Kurnik K, Tretyn A, Tyburski J (2015) The expression patterns of plasma membrane aquaporins in leaves of sugar beet and its halophyte relative, Beta vulgaris ssp. maritima, in response to salt stress. Biologia 70:467–477. https://doi.org/10.1515/biolog-2015-0056
Slama I, Abdelly C, Bouchereau A, Flowers T, Sauvoré A (2015) Diversity, distribution and roles of osmoprotective compounds accumulated in halophytes under abiotic stress. Ann Bot 115:433–447. https://doi.org/10.1093/aob/mcu239
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599. https://doi.org/10.1093/molbev/msm092
Tezara W, Colombo R, Coronel I, Marín O (2011) Water relations and photosynthetic capacity of two species of Calotropis in a tropical semi-arid ecosystem. Annals Botany 107:397–405. https://doi.org/10.1093/aob/mcq245
Thomas RL, Sheard RW, Moyer JR (1967) Comparison of conventional and automated procedures for nitrogen, phosphorus, and potassium analysis of plant material using a single digestion. Agron J 59:240–243. https://doi.org/10.2134/agronj1967.00021962005900030010x
Wallace IS, Roberts DM (2004) Homology modeling of representative subfamilies of Arabidopsis major intrinsic proteins. Classification based on the aromatic/arginine selectivity filter. Plant Physiol 135:1059–1068. https://doi.org/10.1104/pp.103.033415
Xin S, Yu G, Sun L, Qiang X, Xu N, Cheng X (2014) Expression of tomato SlTIP2;2 enhances the tolerance to salt stress in the transgenic Arabidopsis and interacts with target proteins. J Plant Res 127:695–708. https://doi.org/10.1007/s10265-014-0658-7
Yepes-Molina L, Barzana G, Carvajal M (2020) Controversial regulation of gene expression and protein transduction of aquaporins under drought and salinity stress. Plants 9:1662. https://doi.org/10.3390/plants9121662
Zhou L, Wang C, Liu R, Han Q, Vandeleur RK, Du J, Tyerman S (2014) Constitutive overexpression of soybean plasma membrane intrinsic protein GmPIP1;6 confers salt tolerance. BMC Plant Biol 14:181
This work was supported by the National Council for Scientific and Technological Development (CNPq) [CNPq-470247/2013-4; 310871/2014-0; and 433931/2018-3]. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) (Finance Code 001) for scholarship to M.R.C. M.G.S. and A.M.B.I. recognize CNPq for fellowships and financial support. A.M.B.I. recognizes the CAPES BioComputacional Program [88882.160046 / 2013-01] for financial support.
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Coêlho, M.R.V., Rivas, R., Ferreira-Neto, J.R.C. et al. Salt tolerance of Calotropis procera begins with immediate regulation of aquaporin activity in the root system. Physiol Mol Biol Plants (2021). https://doi.org/10.1007/s12298-021-00957-9
- Gas exchange
- Root physiology
- Salt stress
- Tonoplast intrinsic proteins