Abstract
Purpose
The xerophyte Pugionium cornutum is a salt-tolerant species that can accumulate high amounts of Cl− in shoots for osmotic adjustment under saline condition. However, the molecular basis underlying how P. cornutum uses Cl− as a beneficial osmoticum is not clear yet. In this study, we evaluated the function of a chloride channel PcCLCg from P. cornutum in vacuolar Cl− compartmentalization and plant salt tolerance.
Methods
PcCLCg was cloned; its subcellular localization and expression patterns were analyzed; its function in vacuolar Cl− compartmentalization, ion homeostasis and plant salt tolerance were investigated by expression in yeast and Arabidopsis.
Results
PcCLCg was located on the tonoplast, the encoding gene was mainly expressed in shoots, and the expression level was induced by Cl−-salts. The expression of PcCLCg could improve the growth and increase the Cl− content of the yeast mutant Δgef1, in which a chloride channel gene ScGEF1 is deleted, under Cl−-salt treatments. The overexpression of PcCLCg in wild-type Arabidopsis or atclcg mutant alleviated the detrimental effects of NaCl stress on plant growth and significantly increased shoot Cl− and Na+ content under NaCl treatments. Interestingly, PcCLCg-overexpressing lines showed an increased expression of SLAH1 and NHX1 in roots and shoots, respectively, while a decreased expression of SOS1 in roots than wild-type under salt stress.
Conclusions
The upregulated expression of PcCLCg in shoots is conducive to vacuolar Cl− compartmentalization and regulation of Na+ and Cl− accumulation, thus enhancing the osmotic adjustment capacity in the shoot of P. cornutum under saline conditions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
Code availability
Not applicable.
References
Apse MP, Blumwald E (2007) Na+ transport in plants. FEBS Lett 581:2247–2254. https://doi.org/10.1016/j.febslet.2007.04.014
Baetz U, Eisenach C, Tohge T, Martinoia E, De Angeli A (2016) Vacuolar chloride fluxes impact Ion content and distribution during early salinity stress. Plant Physiol 172:1167–1181. https://doi.org/10.1104/pp.16.00183
Barbier-Brygoo H, Vinauger M, Colcombet J, Ephritikhine G, Frachisse J, Maurel C (2000) Anion channels in higher plants: functional characterization, molecular structure and physiological role. Biochem Biophys Acta 1465:199–218. https://doi.org/10.1016/S0005-2736(00)00139-5
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. https://doi.org/10.1046/j.1365-313x.1998.00343.x
Colmenero-Flores JM, Franco-Navarro JD, Cubero-Font P, Peinado-Torrubia P, Rosales MA (2019) Chloride as a beneficial macronutrient in higher plants: new roles and regulation. Int J Mol Sci 20:4686. https://doi.org/10.3390/ijms20194686
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. https://doi.org/10.1186/1746-4811-9-4
Cubero-Font P, Maierhofer T, Jaslan J, Rosales MA, Espartero J, Diaz-Rueda P, Muller HM, Hurter AL, Al-Rasheid KA, Marten I, Hedrich R, Colmenero-Flores JM, Geiger D (2016) Silent S-type anion channel subunit SLAH1 gates SLAH3 open for chloride root-to-shoot translocation. Curr Biol 26:2213–2220. https://doi.org/10.1016/j.cub.2016.06.045
Cui YN, Wang FZ, Yang CH, Yuan JZ, Guo H, Zhang JL, Wang SM, Ma Q (2019) Transcriptomic profiling identifies candidate genes involved in the salt tolerance of the xerophyte Pugionium cornutum. Genes 10:1039. https://doi.org/10.3390/genes10121039
Cui YN, Li XT, Yuan JZ, Wang FZ, Guo H, Xia ZR, Wang SM, Ma Q (2020) Chloride is beneficial for growth of the xerophyte Pugionium cornutum by enhancing osmotic adjustment capacity under salt and drought stresses. J Exp Bot 71:4215–4231. https://doi.org/10.1093/jxb/eraa158
Cui YN, Wang FZ, Yuan JZ, Guo H, Wang SM, Ma Q (2021) High concentrations of sodium and chloride ions have opposing effects on the growth of the xerophyte Pugionium cornutum under saline conditions. J Plant Nutr Soil 184:88–97. https://doi.org/10.1002/jpln.202000148
Cui YN, Li XY, Liu RW, He ZH, Wang SM, Ma Q (2022) SLAH1 is involved in the long-distance transport of Cl- from roots into shoots in the Cl–tolerant xerophyte Pugionium cornutum under salt stress. Plant Soil 479:631–648. https://doi.org/10.1007/s11104-022-05551-w
Davenport RJ, Munoz-Mayor A, Jha D, Essah PA, Rus A, Tester M (2007) The Na+ transporter AtHKT1;1 controls retrieval of Na+ from the xylem in Arabidopsis. Plant Cell Environ 30:497–507. https://doi.org/10.1111/j.1365-3040.2007.01637.x
Drechsler N, Zheng Y, Bohner A, Nobmann B, von Wiren N, Kunze R, Rausch C (2015) Nitrate-dependent control of shoot K homeostasis by the nitrate transporter1/peptide transporter family member NPF7.3/NRT1.5 and the stelar K+ outward rectifier SKOR in Arabidopsis. Plant Physiol 69:2832–2847. https://doi.org/10.1145/2510650.2510660
Duan HR, Ma Q, Zhang JL, Hu J, Bao AK, Wei L, Wang Q, Luan S, Wang SM (2015) The inward-rectifying K+ channel SsAKT1 is a candidate involved in K+ uptake in the halophyte Suaeda salsa under saline condition. Plant Soil 395:173–187. https://doi.org/10.1007/s11104-015-2539-9
Dunkel M, Latz A, Schumacher K, Müller T, Becker D, Hedrich R (2008) Targeting of vacuolar membrane localized members of the TPK channel family. Mol Plant 1:938–949. https://doi.org/10.1093/mp/ssn064
Franco-Navarro JD, Brumós J, Rosales MA, Cubero-Font P, Talón M, Colmenero-Flores JM (2016) Chloride regulates leaf cell size and water relations in tobacco plants. J Exp Bot 67:873–891. https://doi.org/10.1093/jxb/erv502
Franco-Navarro JD, Rosales MA, Cubero-Font P, Calvo P, Álvarez R, Diaz-Espejo A, Colmenero-Flores JM (2019) Chloride as a macronutrient increases water-use efficiency by anatomically driven reduced stomatal conductance and increased mesophyll diffusion to CO2. Plant J 99:815–831. https://doi.org/10.1111/tpj.14423
Gaxiola RA, Yuan DS, Klausner RD, Fink GR (1998) The yeast CLC chloride channel functions in cation homeostasis. Proc Natl Acad Sci USA 95:4046–4050
Gaxiola RA, Rao R, Sherman A, Grisafi P, Alper SL, Fink GR (1999) The Arabidopsis thaliana proton transporters, AtNhx1 and Avp1, can function in cation detoxification in yeast. Proc Natl Acad Sci USA 96:1480–1485. https://doi.org/10.1073/pnas.96.4.1480
Geilfus CM (2018) Chloride: from nutrient to toxicant. Plant Cell Physiol 59:877–886. https://doi.org/10.1093/pcp/pcy071
Hedrich R, Shabala S (2018) Stomata in a saline world. Curr Opin Plant Biol 46:87–95. https://doi.org/10.1016/j.pbi.2018.07.015
Jossier M, Kroniewicz L, Dalmas F, Thiec DL, Ephritikhine G, Barbier-Brygoo H, Vavasseur A, Filleur S, Leonhardt N (2010) The Arabidopsis vacuolar anion transporter, AtCLCc, is involved in the regulation of stomatal movements and contributes to salt tolerance. Plant J 64:563–576. https://doi.org/10.1111/j.1365-313X.2010.04352.x
Li W, Wang L, Cao J, Yu B (2014) Bioinformatics analysis of CLC homologous genes family in soybean genome. J Nanjing Agric Univ 37:35–43. https://doi.org/10.7685/j.issn.1000-2030.2014.03.005
Li H, Li C, Zhang C, Chen B, Hui L, Shen Y (2015) Compositional and gastrointestinal prokinetic studies of Pugionium (L.). Food Chem 186:285–291. https://doi.org/10.1016/j.foodchem.2015.03.146
Li B, Byrt C, Qiu J, Baumann U, Hrmova M, Evrard A, Johnson AA, Birnbaum KD, Mayo GM, Jha D, Henderson SW, Tester M, Gilliham M, Roy SJ (2016) Identification of a stelar-localized transport protein that facilitates root-to-shoot transfer of chloride in Arabidopsis. Plant Physiol 170:1014–1029. https://doi.org/10.1104/pp.15.01163
Li B, Tester M, Gilliham M (2017) Chloride on the move. Trends Plant Sci 22:236–248. https://doi.org/10.1016/j.tplants.2016.12.004
Lv QD, Tang RJ, Liu H, Gao XS, Li YZ, Zheng HQ, Zhang HX (2009) Cloning and molecular analyses of the Arabidopsis thaliana chloride channel gene family. Plant Sci 176:650–661. https://doi.org/10.1016/j.plantsci.2009.02.006
Ma Q, Hu J, Zhou XR, Yuan HJ, Kumar T, Luan S, Wang SM (2017) ZxAKT1 is essential for K+ uptake and K+/Na+ homeostasis in the succulent xerophyte Zygophyllum xanthoxylum. Plant J 90:48–60. https://doi.org/10.1111/tpj.13465
Maresova L, Sychrova H (2006) Arabidopsis thaliana CHX17 gene complements the kha1 deletion phenotypes in Saccharomyces cerevisiae. Yeast 16:1167–1171. https://doi.org/10.1002/yea.1424
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
Munns R, James RA, Xu B, Athman A, Conn SJ, Jordans C, Byrt CS, Hare RA, Tyerman SD, Tester M, Plett D, Gilliham M (2012) Wheat grain yield on saline soils is improved by an ancestral Na+ transporter gene. Nat Biotechnol 30:360–364. https://doi.org/10.1038/nbt.2120
Nguyen CT, Agorio A, Jossier M, Depre S, Thomine S, Filleur S (2016) Characterization of the chloride channel-like, AtCLCg, involved in chloride tolerance in Arabidopsis thaliana. Plant Cell Physiol 57:764–775. https://doi.org/10.1093/pcp/pcv169
Peinado-Torrubia P, Álvarez R, Lucas M, Franco-Navarro JD, Durán-Gutiérrez FJ, Colmenero-Flores JM, Rosales MA (2023) Nitrogen assimilation and photorespiration become more efficient under chloride nutrition as a beneficial macronutrient. Front Plant Sci 13:1058774 (https://www.frontiersin.org/articles/10.3389/fpls.2022.1058774)
Qiu J, Henderson SW, Tester M, Roy SJ, Gilliham M (2016) SLAH1, a homologue of the slow type anion channel SLAC1, modulates shoot Cl- accumulation and salt tolerance in Arabidopsis thaliana. J Exp Bot 67:4495–4505. https://doi.org/10.1093/jxb/erw237
Reich M, Aghajanzadeh T, Helm J, Parmar S, Hawkesford MJ, De Kok LJ (2017) Chloride and sulfate salinity differently affect biomass, mineral nutrient composition and expression of sulfate transport and assimilation genes in Brassica rapa. Plant Soil 411:319–332. https://doi.org/10.1007/s11104-016-3026-7
Rosales MA, Franco-Navarro JD, Peinado-Torrubia P, Díaz-Rueda P, Álvarez R, Colmenero-Flores JM (2020) Chloride improves nitrate utilization and NUE in plants. Front Plant Sci 11:442. https://doi.org/10.3389/fpls.2020.00442
Shabala S (2013) Learning from halophytes: physiological basis and strategies to improve abiotic stress tolerance in crops. Ann Bot 112:1209–1221. https://doi.org/10.1093/aob/mct205
Shi H, Ishitani M, Kim C, Zhu JK (2000) The Arabidopsis thaliana salt tolerance gene SOS1 encodes a putative Na+/H+ antiporter. Proc Natl Acad Sci USA 97:6896–6901. https://doi.org/10.1073/pnas.120170197
Song J, Wang B (2015) Using euhalophytes to understand salt tolerance and to develop saline agriculture: Suaeda salsa as a promising model. Ann Bot 115:541–553. https://doi.org/10.1093/aob/mcu194
Sparkes IA, Runions J, Kearns A, Hawes C (2006) Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 4:2019–2025. https://doi.org/10.1038/nprot.2006.286
Valenzuela FJ, Reineke D, Leventini D, Chen CCL, Barrett-Lennard EG, Colmer TD, Dodd IC, Shabala S, Brown P, Bazihizina N (2022) Plant responses to heterogeneous salinity: agronomic relevance and research priorities. Ann Bot: mcac022. https://doi.org/10.1093/aob/mcac022
Wang SM, Zhang JL, Flowers TJ (2007) Low-affinity Na+ uptake in the halophyte Suaeda maritima. Plant Physiol 145(559–571):781. https://doi.org/10.1104/pp.107.104315
Wang Q, Guan C, Wang P, Lv ML, Ma Q, Wu GQ, Bao AK, Zhang JL, Wang SM (2015) AtHKT1;1 and AtHAK5 mediate low-affinity Na+ uptake in Arabidopsis thaliana under mild salt stress. Plant Growth Regul 75:615–623. https://doi.org/10.1007/s10725-014-9964-2
Wang P, Wang F, Yang J (2017) De novo assembly and analysis of the Pugionium cornutum (L.) Gaertn. transcriptome and identification of genes involved in the drought response. Gene 626:290–297. https://doi.org/10.1016/j.gene.2017.05.053
Wang WY, Liu YQ, Duan HR, Yin XX, Cui YN, Chai WW, Song X, Flowers TJ, Wang SM (2020) SsHKT1;1 is coordinated with SsSOS1 and SsNHX1 to regulate Na+ homeostasis in Suaeda salsa under saline conditions. Plant Soil 449:117–131. https://doi.org/10.1007/s11104-020-04463-x
Wege S, Jossier M, Filleur S, Thomine S, Barbier-Brygoo H, Gambale F, De Angeli A (2010) The proline 160 in the selectivity filter of the Arabidopsis NO3-/H+ exchanger AtCLCa is essential for nitrate accumulation in planta. Plant J 63:861–869. https://doi.org/10.1111/j.1365-313X.2010.04288.x
Wei PP, Wang L, Liu A, Yu B, Lam HM (2016) GmCLC1 confers enhanced salt tolerance through regulating chloride accumulation in soybean. Front Plant Sci 7:1082. https://doi.org/10.3389/fpls.2016.01082
Yu QS, Wang Q, Wang AL, Wu GL, Liu JQ (2010) Interspecific delimitation and phylogenetic origin of Pugionium (Brassicaceae). J Syst Evol 48:195–206. https://doi.org/10.1111/j.1759-6831.2010.00078.x
Yuan HJ, Ma Q, Wu GQ, Wang P, Hu J, Wang SM (2015) ZxNHX controls Na+ and K+ homeostasis at the whole-plant level in Zygophyllum xanthoxylum through feedback regulation of the expression of genes involved in their transport. Ann Bot 115:495–507. https://doi.org/10.1093/aob/mcu177
Yue LJ, Yuan K, Li HW, Kang JJ, Wang SM (2016) Adaptive responses of eremophyte Pugionium cornutum seedlings to different concentrations of NaCl. Acta Prataculturae Sinica 25:144–152. https://doi.org/10.11686/cyxb2015115
Funding
This work was financially supported by the National Natural Science Foundation of China (Grant Nos. 32171677 and 31730093), and Scientific Startup Foundation for Doctors of Northwest A and F University (Grant No. 2452021106).
Author information
Authors and Affiliations
Contributions
YNC and QM conceptualized and designed the study. YNC, RWL and ZRL generated the data. YNC, RWL, ZRL and MMC analyzed the data. YNC and QM wrote the paper, SMW revised the paper.
Corresponding author
Ethics declarations
Ethics approval
Not applicable.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Conflicts of interest/competing interests
The authors declare that they have no conflict of interest.
Additional information
Responsible Editor: Devrim Coskun.
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Cui, YN., Lin, ZR., Cai, MM. et al. PcCLCg is involved in the accumulation of Cl− in shoots for osmotic adjustment and salinity resistance in the Cl−-tolerant xerophyte Pugionium cornutum. Plant Soil 487, 283–298 (2023). https://doi.org/10.1007/s11104-023-05926-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11104-023-05926-7