
CHOLINE TRANSPORTER-RELATED 4 (CTR4) Is Involved in Drought and Saline Tolerance in Rice
Received date: 2024-06-19
Accepted date: 2024-10-14
Online published: 2025-02-20
The tolerance of rice to drought and saline stress is crucial for maintaining yields and promoting widespread cultivation. From an ethyl methanesulfonate (EMS)-mutagenized mutant library, we identified a mutant that is susceptible to osmotic stress, named Osmotic Stress Sensitivity 1 (Oss1). Using MutMap sequencing, we characterized the role of a choline transporter-related family gene, CTR4 (Choline Transporter-Related 4), in rice’s tolerance to drought and salt stress. CTR4 plays a critical role in regulating membrane lipid synthesis. In knockout mutants, the total membrane lipid content, especially unsaturated fatty acids, was significantly reduced. Compared with the wild type, knockout mutants exhibited decreased membrane lipid stability under drought and salt stress, faster water loss, higher relative electrolyte leakage, and lower levels of proline and soluble sugars, leading to impaired tolerance to drought and salt stress. In contrast, the overexpression of CTR4 enhanced seedling tolerance to drought and saline stress. The overexpression lines displayed lower malondialdehyde levels, reduced relative electrolyte leakage, and slower rates of leaf water loss under stress conditions, thereby improving seedling survival rates during stress. Moreover, lipid synthesis gene expression was down-regulated in CTR4 mutants, potentially exacerbating membrane permeability defects and further compromising stress resistance. These findings suggest that CTR4 mediates choline transport and influences cell membrane formation, thereby enhancing rice defenses against drought and salt stress by maintaining lipid homeostasis.
Key words: Oryza sativa; drought stress; salt stress; choline transporter; lipid homeostasis
Yu Shicong, Luo Ruxian, Zheng Shuqin, Ning Jing, Shi Yuanzhu, Guo Daiming, Jia Liangmeng, Wang Sen, Xiao Guizong, Guo Pengwang, Li Yang, Ma Xiaoding . CHOLINE TRANSPORTER-RELATED 4 (CTR4) Is Involved in Drought and Saline Tolerance in Rice[J]. Rice Science, 2025 , 32(1) : 52 -66 . DOI: 10.1016/j.rsci.2024.10.002
| [1] | Apel K, Hirt H. 2004. Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol, 55: 373-399. |
| [2] | Aroca A, Zhang J, Xie Y J, et al. 2021. Hydrogen sulfide signaling in plant adaptations to adverse conditions: Molecular mechanisms. J Exp Bot, 72(16): 5893-5904. |
| [3] | Black S A G, Rylett R J. 2012. Choline transporter CHT regulation and function in cholinergic neurons. Cent Nerv Syst Agents Med Chem, 12(2): 114-121. |
| [4] | Blumwald E, Aharon G S, Apse M P. 2000. Sodium transport in plant cells. Biochim Biophys Acta-Biomembr, 1465(1/2): 140-151. |
| [5] | Botella C, Jouhet J, Block M A. 2017. Importance of phosphatidylcholine on the chloroplast surface. Prog Lipid Res, 65: 12-23. |
| [6] | Chanda B, Xia Y, Mandal M K, et al. 2011. Glycerol-3-phosphate is a critical mobile inducer of systemic immunity in plants. Nat Genet, 43: 421-427. |
| [7] | Chen H X, Li P M, Gao H Y. 2007. Alleviation of photoinhibition by calcium supplement in salt-treated Rumex leaves. Physiol Plant, 129(2): 386-396. |
| [8] | Dettmer J, Ursache R, Campilho A, et al. 2014. CHOLINE TRANSPORTER-LIKE1 is required for sieve plate development to mediate long-distance cell-to-cell communication. Nat Commun, 5: 4276. |
| [9] | Doblas V G, Geldner N, Barberon M. 2017. The endodermis, a tightly controlled barrier for nutrients. Curr Opin Plant Biol, 39: 136-143. |
| [10] | Fu S N, Yang L, Li P, et al. 2011. Aberrant lipid metabolism disrupts calcium homeostasis causing liver endoplasmic reticulum stress in obesity. Nature, 473: 528-531. |
| [11] | Garciadeblás B, Senn M E, Bañuelos M A, et al. 2003. Sodium transport and HKT transporters: The rice model. Plant J, 34(6): 788-801. |
| [12] | Guo R, Yang Z Z, Li F, et al. 2015. Comparative metabolic responses and adaptive strategies of wheat (Triticum aestivum) to salt and alkali stress. BMC Plant Biol, 15: 170. |
| [13] | Hasegawa P M, Bressan R A, Zhu J K, et al. 2000. Plant cellular and molecular responses to high salinity. Annu Rev Plant Physiol Plant Mol Biol, 51: 463-499. |
| [14] | Hong Y Y, Zhao J, Guo L, et al. 2016. Plant phospholipases D and C and their diverse functions in stress responses. Prog Lipid Res, 62: 55-74. |
| [15] | Hu M J, Zhao H M, Yang B, et al. 2021. ZmCTLP1 is required for the maintenance of lipid homeostasis and the basal endosperm transfer layer in maize kernels. New Phytol, 232(6): 2384-2399. |
| [16] | Huang S B, Spielmeyer W, Lagudah E S, et al. 2008. Comparative mapping of HKT genes in wheat, barley, and rice, key determinants of Na+ transport, and salt tolerance. J Exp Bot, 59(4): 927-937. |
| [17] | Jiang Z H, Zhou X P, Tao M, et al. 2019. Plant cell-surface GIPC sphingolipids sense salt to trigger Ca2+ influx. Nature, 572: 341-346. |
| [18] | Kachroo P, Kachroo A. 2020. Lipid-modulated trafficking in plants. Mol Plant, 13(3): 351-353. |
| [19] | Katz-Brull R, Koudinov A R, Degani H. 2002. Choline in the aging brain. Brain Res, 951(2): 158-165. |
| [20] | Kester M, Simonson M S, Mené P, et al. 1989. Interleukin-1 generates transmembrane signals from phospholipids through novel pathways in cultured rat mesangial cells. J Clin Invest, 83(2): 718-723. |
| [21] | Lin C C, Kao C H. 2001. Relative importance of Na+, Cl-, and abscisic acid in NaCl induced inhibition of root growth of rice seedlings. Plant Soil, 237(1): 165-171. |
| [22] | Lin Y C, Kanehara K, Nakamura Y. 2019. Arabidopsis CHOLINE/ ETHANOLAMINE KINASE 1 (CEK1) is a primary choline kinase localized at the endoplasmic reticulum (ER) and involved in ER stress tolerance. New Phytol, 223(4): 1904-1917. |
| [23] | Liu X X, Ma D K, Zhang Z Y, et al. 2019. Plant lipid remodeling in response to abiotic stresses. Environ Exp Bot, 165: 174-184. |
| [24] | Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4): 402-408. |
| [25] | Martínez-Atienza J, Jiang X Y, Garciadeblas B, et al. 2007. Conservation of the salt overly sensitive pathway in rice. Plant Physiol, 143(2): 1001-1012. |
| [26] | Martinière A, Shvedunova M, Thomson A J W, et al. 2011. Homeostasis of plasma membrane viscosity in fluctuating temperatures. New Phytol, 192(2): 328-337. |
| [27] | Michaelson L V, Napier J A, Molino D, et al. 2016. Plant sphingolipids: Their importance in cellular organization and adaption. Biochim Biophys Acta, 1861: 1329-1335. |
| [28] | Michel V, Bakovic M. 2009. The solute carrier 44A1 is a mitochondrial protein and mediates choline transport. FASEB J, 23(8): 2749-2758. |
| [29] | Mittler R. 2002. Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci, 7(9): 405-410. |
| [30] | Mittler R, Vanderauwera S, Gollery M, et al. 2004. Reactive oxygen gene network of plants. Trends Plant Sci, 9(10): 490-498. |
| [31] | Moon S, Kim Y J, Park H E, et al. 2022. OsSNDP3 functions for the polar tip growth in rice pollen together with OsSNDP2, a paralog of OsSNDP3. Rice, 15( 1): 39. |
| [32] | Moradi P, Mahdavi A, Khoshkam M, et al. 2017. Lipidomics unravels the role of leaf lipids in thyme plant response to drought stress. Int J Mol Sci, 18(10): 2067. |
| [33] | Munné-Bosch S, Alegre L. 2002. Interplay between ascorbic acid and lipophilic antioxidant defences in chloroplasts of water- stressed Arabidopsis plants. FEBS Lett, 524: 145-148. |
| [34] | Murphy M P, Holmgren A, Larsson N G, et al. 2011. Unraveling the biological roles of reactive oxygen species. Cell Metab, 13(4): 361-366. |
| [35] | Nakamura T, Fujiwara R, Ishiguro N, et al. 2010. Involvement of choline transporter-like proteins, CTL1 and CTL2, in glucocorticoid- induced acceleration of phosphatidylcholine synthesis via increased choline uptake. Biol Pharm Bull, 33(4): 691-696. |
| [36] | Niewiadomska E, Karpinska B, Romanowska E, et al. 2004. A salinity-induced C3-CAM transition increases energy conservation in the halophyte Mesembryanthemum crystallinum L. Plant Cell Physiol, 45(6): 789-794. |
| [37] | Niu Y, Xiang Y. 2018. An overview of biomembrane functions in plant responses to high-temperature stress. Front Plant Sci, 9: 915. |
| [38] | Qiao K, Wang M, Takano T, et al. 2018. Overexpression of Acyl- CoA-binding protein 1 (ChACBP1) from saline-alkali-tolerant Chlorella sp. enhances stress tolerance in Arabidopsis. Front Plant Sci, 9: 1772. |
| [39] | Shehab G G, Ahmed O K, El-Beltagi H S. 2010. Effects of various chemical agents for alleviation of drought stress in rice plants (Oryza sativa L.). Not Bot Horti Agrobot Cluj-Na, 38(1): 139-148. |
| [40] | Silveira J A G, Viégas R D A, da Rocha I M, et al. 2003. Proline accumulation and glutamine synthetase activity are increased by salt-induced proteolysis in cashew leaves. J Plant Physiol, 160(2): 115-123. |
| [41] | Sofo A, Scopa A, Nuzzaci M, et al. 2015. Ascorbate peroxidase and catalase activities and their genetic regulation in plants subjected to drought and salinity stresses. Int J Mol Sci, 16(6): 13561-13578. |
| [42] | Testerink C, Munnik T. 2005. Phosphatidic acid: A multifunctional stress signaling lipid in plants. Trends Plant Sci, 10(8): 368-375. |
| [43] | Toyooka K, Goto Y, Asatsuma S, et al. 2009. A mobile secretory vesicle cluster involved in mass transport from the Golgi to the plant cell exterior. Plant Cell, 21(4): 1212-1229. |
| [44] | Tuteja N. 2007. Mechanisms of high salinity tolerance in plants. Methods Enzymol, 428: 419-438. |
| [45] | Ueland P M. 2011. Choline and betaine in health and disease. J Inherit Metab Dis, 34(1): 3-15. |
| [46] | van Zelm E, Zhang Y X, Testerink C. 2020. Salt tolerance mechanisms of plants. Annu Rev Plant Biol, 71(1): 403-433. |
| [47] | Wang H, Zhang M S, Guo R, et al. 2012. Effects of salt stress on ion balance and nitrogen metabolism of old and young leaves in rice (Oryza sativa L.). BMC Plant Biol, 12: 194. |
| [48] | Wang J, Nan N, Li N, et al. 2020. A DNA methylation reader- chaperone regulator-transcription factor complex activates OsHKT1;5 expression during salinity stress. Plant Cell, 32(11): 3535-3558. |
| [49] | Wang S M, Wan C G, Wang Y R, et al. 2004. The characteristics of Na+, K+ and free proline distribution in several drought-resistant plants of the Alxa Desert, China. J Arid Environ, 56(3): 525-539. |
| [50] | Wang X M. 2005. Regulatory functions of phospholipase D and phosphatidic acid in plant growth, development, and stress responses. Plant Physiol, 139(2): 566-573. |
| [51] | Wang Y H, Zhang L R, Zhang L L, et al. 2013. A novel stress- associated protein SbSAP14 from Sorghum bicolor confers tolerance to salt stress in transgenic rice. Mol Breed, 32(2): 437-449. |
| [52] | Wang Y J, Zhang X Y, Huang G R, et al. 2020. Dynamic changes in membrane lipid composition of leaves of winter wheat seedlings in response to PEG-induced water stress. BMC Plant Biol, 20(1): 84. |
| [53] | Xu X X, Zhang J J, Yan B W, et al. 2021. The adjustment of membrane lipid metabolism pathways in maize roots under saline-alkaline stress. Front Plant Sci, 12: 635327. |
| [54] | Yamauchi Y, Furutera A, Seki K, et al. 2008. Malondialdehyde generated from peroxidized linolenic acid causes protein modification in heat-stressed plants. Plant Physiol Biochem, 46(8/9): 786-793. |
| [55] | Yu L J, Nie J N, Cao C Y, et al. 2010. Phosphatidic acid mediates salt stress response by regulation of MPK6 in Arabidopsis thaliana. New Phytol, 188(3): 762-773. |
| [56] | Yu Y C, Wang A M, Li X, et al. 2018. Melatonin-stimulated triacylglycerol breakdown and energy turnover under salinity stress contributes to the maintenance of plasma membrane H+-ATPase activity and K+/Na+ homeostasis in sweet potato. Front Plant Sci, 9: 256. |
| [57] | Zhang Q, Liu Y Q, Jiang Y L, et al. 2022. OsASR6 enhances salt stress tolerance in rice. Int J Mol Sci, 23(16): 9340. |
| [58] | Zhu J K. 2016. Abiotic stress signaling and responses in plants. Cell, 167(2): 313-324. |
/
| 〈 |
|
〉 |