陆地棉GDSL脂肪酶基因GhGELP23D对植物细胞伸长发育的影响

doi:10.3864/j.issn.0578-1752.2026.16.003

摘要/Abstract

摘要：

【目的】 GDSL脂肪酶（GDSL esterase/lipase，GELP）家族是一类多功能水解酶，具有广泛的底物识别与催化能力，在植物生长发育中具有重要作用。在棉纤维伸长发育中，陆地棉GDSL脂肪酶基因GhGELP23D显著高表达，分析其影响植物细胞伸长发育的功能，为GELP调控植物细胞发育的机制解析提供一定参考。【方法】以陆地棉纤维为材料克隆GhGELP23D。采用生物信息学方法分析GhGELP23D蛋白的理化性质、结构特征及系统进化关系，利用PlantCARE预测其启动子顺式作用元件，结合公共转录组数据和RT-qPCR数据分析基因的表达特征。构建GhGELP23D启动子GUS报告载体，在拟南芥稳定转化体系中检测GhGELP23D的组织表达特异性。通过烟草叶片瞬时表达系统，对GhGELP23D蛋白进行亚细胞定位。利用花序浸染法获得异源表达GhGELP23D的转基因拟南芥株系GhGELP23D-OE，并分析其表型变化。同时，利用病毒诱导基因沉默（VIGS）技术在陆地棉中沉默GhGELP23D，通过RT-qPCR检测其沉默效率，并观察GhGELP23D-VIGS株系纤维长度的变化。【结果】 GhGELP23D编码一个由356个氨基酸组成的稳定、弱碱性、亲水性蛋白，其结构包含典型信号肽。转录组表达和RT-qPCR数据显示，GhGELP23D在纤维伸长期高表达。亚细胞定位结果表明，GhGELP23D蛋白定位在胞外间隙。启动子组成分析显示，GhGELP23D包含光、激素和胁迫等多种顺式作用元件；组织染色结果显示，GhGELP23D启动子在拟南芥多种组织中均可驱动报告基因表达，表明其具有广泛的组织表达特性。在异源表达GhGELP23D拟南芥中显著促进植株生长发育，GhGELP23D-OE拟南芥株系的株高显著增加，主根和根毛的长度显著提升，在突变体背景中回补表达GhGELP23D后，其主根和根毛长度得到明显恢复并基本达到野生型水平。GhGELP23D-VIGS棉花株系的纤维长度较空载对照植株显著降低，平均下降约13.14%。【结论】 GhGELP23D编码一类定位于胞外间隙的GDSL脂肪酶，在棉纤维伸长期高表达，并影响植物细胞伸长发育过程。

关键词: 陆地棉, GDSL脂肪酶, 细胞伸长, GhGELP23D, 亚细胞定位, 病毒诱导基因沉默

Abstract:

【Objective】 The GDSL esterase/lipase (GELP) family comprises a group of multifunctional hydrolases with broad substrate specificity and catalytic versatility, playing an essential role in plant growth and developmental processes. The GDSL lipase gene GhGELP23D from Gossypium hirsutum is highly expressed during cotton fiber elongation development. This study aims to investigate its function in plant cell elongation and provides references for the mechanism elucidation of GELP-mediated regulation of cell growth. 【Method】 The GhGELP23D was cloned from upland cotton fibers. Bioinformatic analyses were performed to characterize its physicochemical properties, structural features, and phylogenetic relationships. The promoter region was analyzed with PlantCARE to identify putative cis-elements. Expression patterns were detected based on public transcriptomic data and RT-qPCR validation. A GUS reporter construct driven by the GhGELP23D promoter was generated to examine the tissue-specific expression of GhGELP23D in stably transformed Arabidopsis. Subcellular localization of GhGELP23D was determined through transient expression in Nicotiana benthamiana leaves. Transgenic Arabidopsis lines heterologously expressing GhGELP23D (GhGELP23D-OE) were obtained using the floral-dip method, and phenotypic analyses were conducted. Additionally, virus-induced gene silencing (VIGS) was employed to suppress GhGELP23D expression in cotton. Silencing efficiency was verified by RT-qPCR, and the resulting changes in fiber length were assessed in GhGELP23D-VIGS plants. 【Result】 GhGELP23D encodes a 356-amino-acid protein that is stable, weakly basic, and hydrophilic, containing a typical signal peptide but no transmembrane domain. Transcriptomic analysis and RT-qPCR detection showed that GhGELP23D was highly expressed during cotton fiber elongation. Subcellular localization analysis revealed that GhGELP23D is predominantly localized to the extracellular space. Analysis of the GhGELP23D promoter indicated the presence of multiple cis-elements related to light responsiveness, hormone signaling, and stress response. Histochemical GUS staining showed that the GhGELP23D promoter can drive reporter gene expression in various Arabidopsis tissues, indicating broad tissue expression activity. Heterologous expression of GhGELP23D in Arabidopsis (GhGELP23D-OE) significantly enhanced plant growth and development, resulting in increased plant height and longer primary roots and root hairs. Furthermore, complementation of GhGELP23D in the mutant resulted in a pronounced recovery of primary root and root hair length similar to that of the wild type (WT). In cotton, the fiber length of GhGELP23D-VIGS lines was significantly reduced compared with that of empty-vector control plants, with an average decrease of approximately 13.14%.【Conclusion】 GhGELP23D encodes an extracellular GDSL lipase that is highly expressed during the fiber elongation stage of upland cotton and plays an important role in plant cell elongation development.

Key words: Gossypium hirsutum, GDSL lipase, cell elongation, GhGELP23D, subcellular localization, VIGS

陈豪华, 陈港, 冯健庭, 解静, 李倩倩, 王斐, 李鸿彬. 陆地棉GDSL脂肪酶基因GhGELP23D对植物细胞伸长发育的影响[J]. 中国农业科学, 2026, 59(16): 3496-3508.

CHEN HaoHua, CHEN Gang, FENG JianTing, XIE Jing, LI QianQian, WANG Fei, LI HongBin. Effects of the GDSL Lipase Gene GhGELP23D from Gossypium hirsutum on Plant Cell Elongation Development[J]. Scientia Agricultura Sinica, 2026, 59(16): 3496-3508.

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参考文献 36

[1]	Yuan B Z, Sun J. Bibliometric analysis of cotton research from plant sciences category based on web of science[J]. Journal of Natural Fibers, 2022, 19(14): 7579-7593. doi: 10.1080/15440478.2021.1952139
[2]	Huang G, Huang J Q, Chen X Y, Zhu Y X. Recent advances and future perspectives in cotton research[J]. Annual Review of Plant Biology, 2021, 72: 437-462. doi: 10.1146/annurev-arplant-080720-113241 pmid: 33428477
[3]	Kim H J, Triplett B A. Cotton fiber growth in planta and in vitro. Models for plant cell elongation and cell wall biogenesis[J]. Plant Physiology, 2001, 127(4): 1361-1366. pmid: 11743074
[4]	Sun W J, Gao Z Y, Wang J, Huang Y Q, Chen Y, Li J F, Lv M L, Wang J, Luo M, Zuo K J. Cotton fiber elongation requires the transcription factor GhMYB212 to regulate sucrose transportation into expanding fibers[J]. The New Phytologist, 2019, 222(2): 864-881. doi: 10.1111/nph.2019.222.issue-2
[5]	Suo Q W, Fang N J, Zeng J Y, Yan F L, Zhu X, Wang Y, Yu W T, Chen J M, Liang A M, Li Y H, Kong J, Xiao Y H. R2R3 MYB transcription factor GhMYB201 promotes cotton fiber elongation via cell wall loosening and very-long-chain fatty acid synthesis[J]. International Journal of Molecular Sciences, 2024, 25(17): 9559. doi: 10.3390/ijms25179559
[6]	Xiao G H, Zhao P, Zhang Y. A pivotal role of hormones in regulating cotton fiber development[J]. Frontiers in Plant Science, 2019, 10: 87. doi: 10.3389/fpls.2019.00087 pmid: 30838005
[7]	Somssich M, Khan G A, Persson S. Cell wall heterogeneity in root development of Arabidopsis[J]. Frontiers in Plant Science, 2016, 7: 1242. doi: 10.3389/fpls.2016.01242 pmid: 27582757
[8]	Liu X H, Cui H Y, Zhang B C, Song M, Chen S L, Xiao C W, Tang Y J, Liesche J. Reduced pectin content of cell walls prevents stress-induced root cell elongation in Arabidopsis[J]. Journal of Experimental Botany, 2021, 72(4): 1073-1084. doi: 10.1093/jxb/eraa533
[9]	Akoh C C, Lee G C, Liaw Y C, Huang T H, Shaw J F. GDSL family of serine esterases/lipases[J]. Progress in Lipid Research, 2004, 43(6): 534-552. doi: 10.1016/j.plipres.2004.09.002 pmid: 15522763
[10]	Shen G D, Sun W L, Chen Z C, Shi L, Hong J, Shi J X. Plant GDSL esterases/lipases: Evolutionary, physiological and molecular functions in plant development[J]. Plants, 2022, 11(4): 468. doi: 10.3390/plants11040468
[11]	Bischoff V, Nita S, Neumetzler L, Schindelasch D, Urbain A, Eshed R, Persson S, Delmer D, Scheible W R. TRICHOME BIREFRINGENCE and its homolog AT5G01360 encode plant-specific DUF231 proteins required for cellulose biosynthesis in Arabidopsis[J]. Plant Physiology, 2010, 153(2): 590-602. doi: 10.1104/pp.110.153320 pmid: 20388664
[12]	Ji R, Wang H, Xin X, Peng S, Hur Y, Li Z, Feng H. BrEXL6, a GDSL lipase gene of Brassica rapa, functions in pollen development[J]. Biologia Plantarum, 2017, 61(4): 685-692. doi: 10.1007/s10535-017-0735-6
[13]	Zhang B C, Zhang L J, Li F, Zhang D M, Liu X L, Wang H, Xu Z P, Chu C C, Zhou Y H. Control of secondary cell wall patterning involves xylan deacetylation by a GDSL esterase[J]. Nature Plants, 2017, 3: 17017. doi: 10.1038/nplants.2017.17 pmid: 28260782
[14]	孙英超, 刘伟, 何秋伶, 祝水金, 陈进红. 不同脂肪酸种类及浓度对棉纤维分化发育的影响[J]. 浙江大学学报(农业与生命科学版), 2014, 40(6): 621-626.
	Sun Y C, Liu W, He Q L, Zhu S J, Chen J H. Effects of different kinds and concentrations of fatty acids on differentiation and development of cotton fiber[J]. Journal of Zhejiang University (Agriculture and Life Sciences), 2014, 40(6): 621-626. (in Chinese)
[15]	梅文倩, 秦咏梅, 朱玉贤. 乙烯、超长链脂肪酸、活性氧、油菜素内酯和赤霉素相互作用调控棉纤维伸长发育的分子机制研究[J]. 生命科学, 2010, 22(1): 7-14.
	Mei W Q, Qin Y M, Zhu Y X. Cross-talk among ethylene, very long-chain fatty acids, reactive oxygen species, brassinosteroid and gibberellin mediates cotton fiber elongation[J]. Chinese Bulletin of Life Sciences, 2010, 22(1): 7-14. (in Chinese)
[16]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C(T)) Method[J]. Methods, 2001, 25(4): 402-408. doi: 10.1006/meth.2001.1262 pmid: 11846609
[17]	Fernandez-Pozo N, Rosli H G, Martin G B, Mueller L A. The SGN VIGS tool: User-friendly software to design virus-induced gene silencing (VIGS) constructs for functional genomics[J]. Molecular Plant, 2015, 8(3): 486-488. doi: 10.1016/j.molp.2014.11.024 pmid: 25667001
[18]	Zhang X R, Henriques R, Lin S S, Niu Q W, Chua N H. Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method[J]. Nature Protocols, 2006, 1(2): 641-646. doi: 10.1038/nprot.2006.97
[19]	Oh I S, Park A R, Bae M S, Kwon S J, Kim Y S, Lee J E, Kang N Y, Lee S, Cheong H, Park O K. Secretome analysis reveals an Arabidopsis lipase involved in defense against Alternaria brassicicola[J]. The Plant Cell, 2005, 17(10): 2832-2847. doi: 10.1105/tpc.105.034819
[20]	李志兰, 华水金, 蒋立希. 植物GDSL脂酶家族基因的研究进展[J]. 农业生物技术学报, 2014, 22(7): 916-924.
	Li Z L, Hua S J, Jiang L X. Advances in studying plant GDSL-type lipase genes[J]. Journal of Agricultural Biotechnology, 2014, 22(7): 916-924. (in Chinese)
[21]	周丽霞, 杨蒙迪, 李睿. 油棕GDSL酯酶/脂肪酶基因的全基因组鉴定及表达分析[J]. 分子植物育种, 2023, 21(14): 4545-4559.
	Zhou L X, Yang M D, Li R. Genome-wide identification and expression analysis of GDSL esterasr/lipase genes in Elaeis guineensis[J]. Molecular Plant Breeding, 2023, 21(14): 4545-4559. (in Chinese)
[22]	奈婕菲, 程玉祥. 一个杨树GDSL基因组织表达的特性及其在拟南芥异源的表达[J]. 江苏农业科学, 2014, 42(3): 16-18.
	Nai J F, Cheng Y X. Characteristics of tissue expression of a poplar GDSL gene and its heterologous expression in Arabidopsis thaliana[J]. Jiangsu Agricultural Sciences, 2014, 42(3): 16-18. (in Chinese)
[23]	Takahashi K, Shimada T, Kondo M, Tamai A, Mori M, Hara-Nishimura I. Ectopic expression of an esterase, which is a candidate for the unidentified plant cutinase, causes cuticular defects in Arabidopsis thaliana[J]. Plant & Cell Physiology, 2010, 51(1): 123-131.
[24]	Updegraff E P, Zhao F, Preuss D. The extracellular lipase EXL4 is required for efficient hydration of Arabidopsis pollen[J]. Sexual Plant Reproduction, 2009, 22(3): 197-204. doi: 10.1007/s00497-009-0104-5 pmid: 20033440
[25]	Wang Z Y, Cai C P, Yu Z H, Reyimu Y, Han X, Lv M, Zhang Z H, Li W X, Zhu G Z, Guo W Z. A GDSL esterase/lipase gene GbGELP identified from a fiber micronaire QTL qMIC-A11 modulates cell elongation and fiber development[J]. Theoretical and Applied Genetics, 2025, 138(6): 126. doi: 10.1007/s00122-025-04915-x pmid: 40413700
[26]	Serna L, Martin C. Trichomes: Different regulatory networks lead to convergent structures[J]. Trends in Plant Science, 2006, 11(6): 274-280. doi: 10.1016/j.tplants.2006.04.008 pmid: 16697247
[27]	Jiao Y, Zhang J H, Pan C D. Genome-wide analysis of the GDSL genes in pecan (Carya illinoensis K. Koch): Phylogeny, structure, promoter Cis-elements, co-expression networks, and response to salt stresses[J]. Genes, 2022, 13(7): 1103. doi: 10.3390/genes13071103
[28]	Rastogi L, Chaudhari A A, Sharma R, Pawar P A. Arabidopsis GELP7 functions as a plasma membrane-localized acetyl xylan esterase, and its overexpression improves saccharification efficiency[J]. Plant Molecular Biology, 2022, 109(6): 781-797. doi: 10.1007/s11103-022-01275-8 pmid: 35577991
[29]	Qin Y M, Hu C Y, Pang Y, Kastaniotis A J, Hiltunen J K, Zhu Y X. Saturated very-long-chain fatty acids promote cotton fiber and Arabidopsis cell elongation by activating ethylene biosynthesis[J]. The Plant Cell, 2007, 19(11): 3692-3704. doi: 10.1105/tpc.107.054437
[30]	Huang L M, Lai C P, Chen L O, Chan M T, Shaw J F. Arabidopsis SFAR4 is a novel GDSL-type esterase involved in fatty acid degradation and glucose tolerance[J]. Botanical Studies, 2015, 56(1): 33. doi: 10.1186/s40529-015-0114-6
[31]	Liu G J, Xiao G H, Liu N J, Liu D, Chen P S, Qin Y M, Zhu Y X. Targeted lipidomics studies reveal that linolenic acid promotes cotton fiber elongation by activating phosphatidylinositol and phosphatidylinositol monophosphate biosynthesis[J]. Molecular Plant, 2015, 8(6): 911-921. doi: 10.1016/j.molp.2015.02.010
[32]	Zhang Y J, Bai B, Lee M, Alfiko Y, Suwanto A, Yue G H. Cloning and characterization of EgGDSL, a gene associated with oil content in oil palm[J]. Scientific Reports, 2018, 8: 11406. doi: 10.1038/s41598-018-29492-6 pmid: 30061629
[33]	Lu R, Li Y, Zhang J, Wang Y, Zhang J, Li Y, Zheng Y, Li X B. The bHLH/HLH transcription factors GhFP2 and GhACE1 antagonistically regulate fiber elongation in cotton[J]. Plant Physiology, 2022, 189(2): 628-643. doi: 10.1093/plphys/kiac088 pmid: 35226094
[34]	Wang N N, Li Y, Chen Y H, Lu R, Zhou L, Wang Y, Zheng Y, Li X B. Phosphorylation of WRKY16 by MPK3-1 is essential for its transcriptional activity during fiber initiation and elongation in cotton (Gossypium hirsutum)[J]. The Plant Cell, 2021, 33(8): 2736-2752. doi: 10.1093/plcell/koab153
[35]	Jin F, Zhu L P, Hou L Y, Li H B, Li L, Xiao G H. Auxin resistant 2 and short hypocotyl 2 regulate cotton fiber initiation and elongation[J]. Plant Physiology, 2024, 195(3): 2032-2052. doi: 10.1093/plphys/kiae183 pmid: 38527791
[36]	Tian Z L, Zhang Y Z, Zhu L P, Jiang B, Wang H Q, Gao R X, Friml J, Xiao G H. Strigolactones act downstream of gibberellins to regulate fiber cell elongation and cell wall thickness in cotton (Gossypium hirsutum)[J]. The Plant Cell, 2022, 34(12): 4816-4839. doi: 10.1093/plcell/koac270 pmid: 36040191