Synergistic role of GhGCS1 in cotton root development and verticillium wilt resistance

Abstract

Sphingolipids are not only essential structural components of cellular membranes but also key signaling molecules that regulate plant growth, development, and stress responses. However, their specific roles in root development remain largely unknown. In the presented study, we demonstrate that overexpression of the GhGCS1 gene significantly enhances lateral root development in cotton. Integrated comprehensive transcriptome analysis and phytohormone quantification revealed that GhGCS1 promotes lateral root initiation and elongation primarily by suppressing cytokinin biosynthesis. Notably, GhGCS1 overexpression also markedly improved cotton resistance to Verticillium dahliae. Further molecular analyses indicated that GhGCS1 enhances cotton verticillium wilt resistance through modulation of the expression of sphingolipid-associated brassinosteroid- and pathogenesis-related genes. Collectively, these findings reveal a dual regulatory role for GhGCS1 in coordinating root architecture and immune responses in cotton, providing novel insights and potential strategies for developing crop varieties with improved yield potential and stress tolerance.

Keywords

Cotton / Sphingolipid / Root development / Verticillium wilt

References

[1] Ali U, Li H, Wang X, Guo L. Emerging roles of sphingolipid signaling in plant response to biotic and abiotic stresses. Mol Plant, 2018, 11(11): 1328-1343

[2] Altamura MM, Piacentini D, Della Rovere F, Fattorini L, Falasca G, Betti C. New paradigms in brassinosteroids, strigolactones, sphingolipids, and nitric oxide interaction in the control of lateral and adventitious root formation. Plants, 2023

[3] Breslow DK, Weissman JS. Membranes in balance: mechanisms of sphingolipid homeostasis. Mol Cell, 2010, 40(2): 267-279

[4] Cacas J-L, Buré C, Grosjean K, Gerbeau-Pissot P, Lherminier J, Rombouts Y, Maes E, Bossard C, Gronnier J, Furt F, Fouillen L, Germain V, Bayer E, Cluzet S, Robert F, Schmitter J-M, Deleu M, Lins L, Simon-Plas F, Mongrand S. Revisiting plant plasma membrane lipids in tobacco: a focus on sphingolipids. Plant Physiol, 2016, 170(1): 367-384

[5] Chen M, Han G, Dietrich CR, Dunn TM, Cahoon EB. The essential nature of sphingolipids in plants as revealed by the functional identification and characterization of theArabidopsisLCB1 subunit of serine palmitoyltransferase. Plant Cell, 2006, 18(12): 3576-3593

[6] Chen M, Markham JE, Dietrich CR, Jaworski JG, Cahoon EB. Sphingolipid long-chain base hydroxylation is important for growth and regulation of sphingolipid content and composition inArabidopsis. Plant Cell, 2008, 20(7): 1862-1878

[7] Chen M, Markham JE, Cahoon EB. Sphingolipid Δ8 unsaturation is important for glucosylceramide biosynthesis and low-temperature performance in Arabidopsis. Plant J, 2011, 69(5): 769-781

[8] Chen Q, Xu F, Wang L, Suo X, Wang Q, Meng Q, Huang L, Ma C, Li G, Luo M. Sphingolipid profile during cotton fiber growth revealed that a phytoceramide containing hydroxylated and saturated VLCFA is important for fiber cell elongation. Biomolecules, 2021

[9] Derbyshire MC, Newman TE, Thomas WJW, Batley J, Edwards D. The complex relationship between disease resistance and yield in crops. Plant Biotechnol J, 2024, 22(9): 2612-2623

[10] Gao S, Fang J, Xu F, Wang W, Sun X, Chu J, Cai B, Feng Y, Chu C. CYTOKININ OXIDASE/DEHYDROGENASE4 integrates cytokinin and auxin signaling to control rice crown root formation. Plant Physiol, 2014, 165(3): 1035-1046

[11] Gao Y, Liu F, Zhang J, Qiao Z, Tan P, Liu N, Hu M, Zhang J, Yan X, Yi Z, Luo M, Xu F. Saturated ceramide is required for seed germination in soybean. Seed Biol, 2025, 4(1): 0-0

[12] Gömann J, Herrfurth C, Zienkiewicz A, Ischebeck T, Haslam TM, Hornung E, Feussner I. Sphingolipid long-chain base hydroxylation influences plant growth and callose deposition in Physcomitrium patens. New Phytol, 2021, 231(1): 297-314

[13] Haslam TM, Feussner I, Nakamura Y. Diversity in sphingolipid metabolism across land plants. J Exp Bot, 2022, 73(9): 2785-2798

[14] Huang G, Huang J-Q, Chen X-Y, Zhu Y-X. Recent advances and future perspectives in cotton research. Annu Rev Plant Biol, 2021, 72(1): 437-462

[15] Huby E, Napier JA, Baillieul F, Michaelson LV, Dhondt‐Cordelier S. Sphingolipids: towards an integrated view of metabolism during the plant stress response. New Phytol, 2019, 225(2): 659-670

[16] Jiang Z, Zhou X, Tao M, Yuan F, Liu L, Wu F, Wu X, Xiang Y, Niu Y, Liu F, Li C, Ye R, Byeon B, Xue Y, Zhao H, Wang H-N, Crawford BM, Johnson DM, Hu C, Pei C, Zhou W, Swift GB, Zhang H, Vo-Dinh T, Hu Z, Siedow JN, Pei Z-M. Plant cell-surface GIPC sphingolipids sense salt to trigger Ca2+ influx. Nature, 2019, 572(7769): 341-346

[17] Karlova R, Boer D, Hayes S, Testerink C. Root plasticity under abiotic stress. Plant Physiol, 2021, 187(3): 1057-1070

[18] Krüger F, Krebs M, Viotti C, Langhans M, Schumacher K, Robinson DG. PDMP induces rapid changes in vacuole morphology in Arabidopsis root cells. J Exp Bot, 2013, 64(2): 529-540

[19] Lenarčič TAI, Böhm H, Hodnik V, Pirc K, Zavec AB, Podobnik M, Pahovnik D, Žagar E, Pruitt R, Greimel P, Yamaji-Hasegawa A, Kobayashi T, Zienkiewicz A, Gömann J, Mortimer JC, Fang L, Mamode-Cassim A, Deleu M, Lins L, Oecking C, Feussner I, Mongrand S, Anderluh G, Nürnberger T. Eudicot plant-specific sphingolipids determine host selectivity of microbial NLP cytolysins. Science, 2017, 358(6369): 1431-1434

[20] Li H, Dai J, Qin J, Shang W, Chen J, Zhang L, Dai X, Klosterman SJ, Xu X, Subbarao KV, Fan S, Hu X. Genome sequences of Verticillium dahliae defoliating strain XJ592 and nondefoliating strain XJ511. Mol Plant Microbe Interact®, 2020, 33(4): 565-568

[21] Li X, Yu J, Zhou W, Yan F, Lin C, Tao Z. A homeobox transcription factor HB34 suppresses jasmonic acid biosynthesis but promotes the expression of growth-related genes to balance plant immunity and growth in Arabidopsis. Plant Commun, 2025

[22] Li Y-K, Zhang Y-M, Dai G-Y, Chen Y-L, Chen D-K, Yao N. Sphingolipid remodeling in the plasma membrane is essential for osmotic stress tolerance in Arabidopsis. Plant Physiol, 2025

[23] Li G, Wang Q, Meng Q, Wang G, Xu F, Chen Q, Liu F, Hu Y, Luo M (2022) Overexpression of a ceramide synthase gene,GhCS1, inhibits fiber cell initiation and elongation by promoting the synthesis of ceramides containing dihydroxy LCB and VLCFA. Front Plant Sci 13:1000348. https://doi.org/10.3389/fpls.2022.1000348

[24] Lin Y, Zhu Y, Wang L, Zheng Y, Xie Y, Cai Q, He W, Xie H, Liu H, Wang Y, Cui L, Wei Y, Xie H, Zhang J (2023) Overexpression of a GIPC glycosyltransferase gene, OsGMT1, suppresses plant immunity and delays heading time in rice. Plant Sci 331:111674. https://doi.org/10.1016/j.plantsci.2023.111674

[25] Liu N-J, Zhang T, Liu Z-H, Chen X, Guo H-S, Ju B-H, Zhang Y-Y, Li G-Z, Zhou Q-H, Qin Y-M, Zhu Y-X. Phytosphinganine affects plasmodesmata permeability via facilitating PDLP5-stimulated callose accumulation in Arabidopsis. Mol Plant, 2020, 13(1): 128-143

[26] Liu N-J, Hou L-P, Bao J-J, Wang L-J, Chen X-Y. Sphingolipid metabolism, transport, and functions in plants: recent progress and future perspectives. Plant Commun, 2021

[27] Lou H, Li S, Shi Z, Zou Y, Zhang Y, Huang X, Yang D, Yang Y, Li Z, Xu C. Engineering source-sink relations by prime editing confers heat-stress resilience in tomato and rice. Cell, 2025, 188(2): 530-549.e520

[28] Ma Y, Xu J, Qi J, Zhao D, Jin M, Wang T, Yang Y, Shi H, Guo L, Zhang H. Crosstalk among plant hormone regulates the root development. Plant Signal Behav, 2024

[29] Ma Y, Zhang Y, Xu J, Qi J, Liu X, Guo L, Zhang H. Research on the mechanisms of phytohormone signaling in regulating root development. Plants, 2024

[30] Meng Q, Wang Q, Xu F, Chen Q, Ma C, Huang L, Li G, Liu F, Luo M (2022) Down-regulating a fiber-specific KCR like gene GhKCRL1 suppressed fiber elongation through blocking the synthesis of sphingolipids in fiber cell. Ind Crops Prod 186:115290. https://doi.org/10.1016/j.indcrop.2022.115290

[31] Moreno-Risueno MA, Van Norman JM, Moreno A, Zhang J, Ahnert SE, Benfey PN. Oscillating gene expression determines competence for periodic Arabidopsis root branching. Science, 2010, 329(5997): 1306-1311

[32] Ouyang Z, Tan Z, Ali U, Zhang Y, Li B, Yao X, Yang B, Guo L. Ceramide-1-phosphate enhances defense responses against Sclerotinia sclerotiorum in Brassica napus. Plant Physiol, 2025

[33] Petricka JJ, Winter CM, Benfey PN. Control of Arabidopsis root development. Annu Rev Plant Biol, 2012, 63(1): 563-590

[34] Rugen MD, Vernet MMJL, Hantouti L, Soenens A, Andriotis VME, Rejzek M, Brett P, van den Berg RJBHN, Aerts JMFG, Overkleeft HS, Field RA (2018) A chemical genetic screen reveals that iminosugar inhibitors of plant glucosylceramide synthase inhibit root growth in Arabidopsis and cereals. Sci Rep 8(1):16421. https://doi.org/10.1038/s41598-018-34749-1

[35] Ryan PR, Liu Q, Sperling P, Dong B, Franke S, Delhaize E. A higher plant Δ8 sphingolipid desaturase with a preference for (Z)-isomer formation confers aluminum tolerance to yeast and plants. Plant Physiol, 2007, 144(4): 1968-1977

[36] Wang JZL, Shi H, Chern M, Yu H, Yi H, He M, Yin J, Zhu X, Li Y, Li W, Liu J, Wang J, Chen X, Qing H, Wang Y, Liu G, Wang W, Li P, Wu X, Zhu L, Zhou JM, Ronald PC, Li S, Li J, Chen X. A single transcription factor promotes both yield and immunity in rice. Science, 2018, 361(6406): 1026-1028

[37] Wang L, Liu C, Liu Y, Luo M. Fumonisin B1-induced changes in cotton fiber elongation revealed by sphingolipidomics and proteomics. Biomolecules, 2020

[38] Wang Q, Yan X, Niu Q, Zhang H, Tian H, Chen Z, Xu F, You Y, Gao Y, Qiao Z, Luo M. Glucosylceramides containing very long-acyl-chain fatty acid are critical for cotton fiber elongation by influencing brassinosteroid synthesis and signaling. Crop J, 2025

[39] Wang X, Zhang Z, Peng W, Huang J, Yan X, Yao W, Ouyang J, Li S (2023) Inositolphosphorylceramide synthases, OsIPCSs, regulate plant height in rice. Plant Sci 335:111798. https://doi.org/10.1016/j.plantsci.2023.111798

[40] Wen X, Chen Z, Yang Z, Wang M, Jin S, Wang G, Zhang L, Wang L, Li J, Saeed S, He S, Wang Z, Wang K, Kong Z, Li F, Zhang X, Chen X, Zhu Y. A comprehensive overview of cotton genomics, biotechnology and molecular biological studies. Sci China Life Sci, 2023, 66(10): 2214-2256

[41] Xu F, Huang L, Wang J, Ma C, Tan Y, Wang F, Fan Y, Luo M. Sphingolipid synthesis inhibitor fumonisin B1 causes verticillium wilt in cotton. J Integr Plant Biol, 2022, 64(4): 836-842

[42] Xu F, Wang L, Xu J, Chen Q, Ma C, Huang L, Li G, Luo M. GhIQD10 interacts with GhCaM7 to control cotton fiber elongation via calcium signaling. Crop J, 2023, 11(2): 447-456

[43] Xu F, Li G, He S, Zeng Z, Wang Q, Zhang H, Yan X, Hu Y, Tian H, Luo M. Sphingolipid inhibitor response gene GhMYB86 controls fiber elongation by regulating microtubule arrangement. J Integr Plant Biol, 2024, 66(9): 1898-1914

[44] Yang B, Peng Y, Zhang G, Liu R, Hao S, Ren Y, Lu S, Wang X, Guo L. Regulation of glucosylceramide synthase and sphingolipid remodeling in the plant response to phosphate deficiency. Plant Cell, 2025

[45] Yang C, Lai Y-M, Yao N (2025b) Plant sphingolipids: Subcellular distributions and functions. Curr Opin Plant Biol 85:102704. https://doi.org/10.1016/j.pbi.2025.102704

[46] Zeng H-Y, Li C-Y, Yao N (2020) Fumonisin B1: A Tool for Exploring the Multiple Functions of Sphingolipids in Plants. Front Plant Sci 11:600458. https://doi.org/10.3389/fpls.2020.600458

[47] Zhang J, Meng Q, Wang Q, Zhang H, Tian H, Wang T, Xu F, Yan X, Luo M (2024) Cotton sphingosine kinase GhLCBK1 participates in fiber cell elongation by affecting sphingosine-1-phophate and auxin synthesis. Int J Biol Macromol 267(Pt 1):131323. https://doi.org/10.1016/j.ijbiomac.2024.131323

Funding

National Natural Science Foundation of China(32372114)

RIGHTS & PERMISSIONS

The Author(s)