黄瓜CsAuxin-15A的克隆及其在盐碱胁迫下的功能验证

doi:10.14088/j.cnki.issn0439-8114.2026.06.036

摘要/Abstract

摘要： 为探究SAUR（Small auxin up-regulated RNA）家族基因CsAuxin-15A在黄瓜（Cucumis sativus L.）响应盐碱胁迫中的生物学功能,以耐盐碱黄瓜品系D1909和盐碱敏感品系D1604为材料,克隆CsAuxin-15A基因,并对其进行生物信息学分析、亚细胞定位、启动子顺式作用元件预测及表达模式分析,同时构建正义和反义表达载体转入黄瓜,测定转基因植株在盐碱胁迫下的过氧化物酶（POD）活性和脯氨酸（Pro）含量。结果表明,CsAuxin-15A全长303 bp,编码100个氨基酸,属于SAUR型生长素反应蛋白家族,定位于细胞质;其启动子区域含有光响应、脱落酸响应、防御与应激反应、ARE厌氧诱导及MYB/MYC结合位点等多种顺式作用元件。盐碱胁迫下,CsAuxin-15A在D1909的茎、叶片和果实中的相对表达量显著高于对照,在叶片中表达量最高,而在根中无显著差异。与野生型植株相比,转正义CsAuxin-15A植株在盐碱胁迫下生长状况更优,叶片过氧化物酶活性和脯氨酸含量显著高于对照。综上,CsAuxin-15A通过正调控抗氧化酶活性和渗透调节物质的积累,增强黄瓜对盐碱胁迫的耐受性。

关键词: 黄瓜（Cucumis sativus L.）, CsAuxin-15A, 克隆, 盐碱胁迫, 功能验证

Abstract: To investigate the biological function of the SAUR（Small auxin up-regulated RNA）family gene CsAuxin-15A in cucumber（Cucumis sativus L.）in response to saline-alkali stress, the CsAuxin-15A gene was cloned from the saline-alkali tolerant cucumber line D1909 and the saline-alkali sensitive line D1604, bioinformatics analysis, subcellular localization, prediction of cis-acting elements in the promoter and expression pattern analysis were performed, sense and antisense expression vectors were simultaneously constructed to transform cucumber, and the peroxidase（POD）activity and proline（Pro）content of transgenic plants under saline-alkali stress were measured. The results showed that the full-length of CsAuxin-15A was 303 bp, encoding 100 amino acids, belonged to the SAUR-type auxin-responsive protein family, and was localized in the cytoplasm; its promoter region contained multiple cis-acting elements such as light-responsive, abscisic acid-responsive, defense and stress-responsive, ARE anaerobic induction, and MYB/MYC binding sites. Under saline-alkali stress, the relative expression of CsAuxin-15A in stems, leaves and fruits of D1909 was significantly higher than that in the control, with the highest expression in leaves, while no significant difference was observed in roots. Compared with wild-type plants, the CsAuxin-15A sense-transgenic plants grew better under saline-alkali stress, and the leaf peroxidase activity and proline content were significantly higher than those in the control. In conclusion, CsAuxin-15A enhanced the tolerance of cucumber to saline-alkali stress by positively regulating the antioxidant enzyme activity and the accumulation of osmotic adjustment substances.

Key words: Cucumis sativus L., CsAuxin-15A, cloning, saline-alkali stress, functional verification

中图分类号:

S642.2

杨洋, 张君鸣, 于聪睿, 张继豪, 罗天佑, 张帆. 黄瓜CsAuxin-15A的克隆及其在盐碱胁迫下的功能验证[J]. 湖北农业科学, 2026, 65(6): 234-245.

YANG Yang, ZHANG Jun-ming, YU Cong-rui, ZHANG Ji-hao, LUO Tian-you, ZHANG Fan. Cloning and functional verification of cucumber CsAuxin-15A under saline-alkali stress[J]. HUBEI AGRICULTURAL SCIENCES, 2026, 65(6): 234-245.

参考文献

[1] 胡平根,张勇涛,齐子旺.黄瓜种植及管理环节相关技术的科学应用与实践探讨[J].种子世界,2025(12):45-47.
[2] 聂圣贤. 黄瓜耐盐砧木筛选及其应用[D].南宁:广西大学,2023.
[3] 申佳丽,魏彦凤,丁增伟,等.蚯蚓粪与脱硫石膏配施对设施黄瓜连作障碍的效应研究[J].核农学报,2024,38(6):1163-1174.
[4] 吴淑杭,周德平,姜震方.盐碱地改良与利用研究进展[J].上海农业科技,2007(2):23-25.
[5] 刘莹. 大庆地区土壤盐碱化成因及改良对策[J].黑龙江水利科技,2015,43(6):151-152.
[6] 李艳茹,司龙亭.黄瓜幼苗耐盐性的遗传分析[J].西北农林科技大学学报(自然科学版),2011,39(4):127-131.
[7] LIU D R,DONG S Y, BO K L, et al.Identification of QTLs controlling salt tolerance in cucumber (Cucumis sativus L.) seedlings[J]. Plants, 2021, 10(1): 85.
[8] 沈建兰. 外源腐胺调节盐胁迫下黄瓜幼苗叶片淀粉积累的机制[D].南京:南京农业大学,2019.
[9] NIU M L, HUANG Y, SUN S T, et al.Root respiratory burst oxidase homologue-dependent H₂O₂ production confers salt tolerance on a grafted cucumber by controlling Na+ exclusion and stomatal closure[J]. Journal of experimental botany,2018,69(14):3465-3476.
[10] GÉVAUDANT F, DUBY G, VON STEDINGK E, et al. Expression of a constitutively activated plasma membrane H+-ATPase alters plant development and increases salt tolerance[J]. Plant physiology, 2007, 144(4): 1763-1776.
[11] WANG Y N, LI K X, LI X.Auxin redistribution modulates plastic development of root system architecture under salt stress in Arabidopsis thaliana[J]. Journal of plant physiology, 2009, 166(15): 1637-1645.
[12] 李红,李波,杨曌.外源ABA对苏打盐碱胁迫的紫花苜蓿内源激素含量的影响[J].黑龙江畜牧兽医,2020(6):103-106,111.
[13] BAO D F, CHANG S Q, LI X D, et al.Advances in the study of auxin early response genes: Au_x/IAA GH3 and SAUR[J]. The crop journal, 2024, 12(4): 964-978.
[14] 翟含含,翟宇杰,田义,等.桃SAUR家族基因分析及PpSAUR5功能鉴定[J].园艺学报,2023,50(1):1-14.
[15] REN H, GRAY W M.SAUR proteins as effectors of hormonal and environmental signals in plant growth[J]. Molecular plant, 2015, 8(8): 1153-1164.
[16] 何高镜. 马铃薯SAUR基因家族分析及StSAUR46基因功能的初步研究[D].长沙:湖南农业大学,2022.
[17] QIU T, QI M Y, DING X H, et al.The saur41 subfamily of small auxin up RNA genes is abscisic acid inducible to modulate cell expansion and salt tolerance in Arabidopsis thaliana seedlings[J]. Annals of botany, 2020, 125(5): 805-819.
[18] 李风光. 桑树生长素早期响应基因Mul-SAUR15的功能研究[D].山东泰安:山东农业大学,2020.
[19] LIU R, WEN S S, SUN T T, et al.PagWOX11/12a positively regulates the PagSAUR36 gene that enhances adventitious root development in poplar[J]. Journal of experimental botany, 2022, 73(22): 7298-7311.
[20] 吴春梅. 拟南芥生长素响应基因SAUR6、SAUR14和SAUR16对抗冻的作用和机理研究[D].兰州:兰州大学,2021.
[21] 李奇峰. 基于多组学筛选黄瓜耐盐碱候选基因[D].黑龙江大庆:黑龙江八一农垦大学,2025.
[22] 韩毅强,高亚梅,杜艳丽,等.大豆耐盐碱种质资源鉴定[J].中国油料作物学报,2021,43(6):1016-1024.
[23] WAN H J, ZHAO Z G, QIAN C T, et al.Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber[J]. Analytical biochemistry, 2010, 399(2): 257-261.
[24] SCHMITTGEN T D, LIVAK K J.Analyzing real-time PCR data by the comparative CT method[J]. Nature protocols, 2008, 3(6): 1101-1108.
[25] YOO S D, CHO Y H, SHEEN J.Arabidopsis mesophyll protoplasts: A versatile cell system for transient gene expression analysis[J]. Nature protocols, 2007, 2(7): 1565-1572.
[26] CHEN S B, SONGKUMARN P, LIU J L, et al.A versatile zero background T-vector system for gene cloning and functional genomics[J]. Plant physiology, 2009, 150(3): 1111-1121.
[27] ZHAO S J, CANG J.Guidance for plant physiology experiments[M]. Beijing, China:China agriculture press, 2016.
[28] STRIMBECK G R, SCHABERG P G, FOSSDAL C G, et al.Extreme low temperature tolerance in woody plants[J]. Frontiers in plant science, 2015, 6: 884.
[29] MENSINK M A,FRIJLINK H W,VAN DER VOORT MAARSCHALK K, et al. How sugars protect proteins in the solid state and during drying (review): Mechanisms of stabilization in relation to stress conditions[J]. European journal of pharmaceutics and biopharmaceutics, 2017, 114: 288-295.
[30] 邹文会,苏亚春,任永娟,等.植物MYC转录因子:结构特点、作用机制和功能调控[J].农业生物技术学报,2022,30(6):1186-1201.
[31] 安飞飞,罗秀芹,蔡杰,等.木薯MeMYB106基因克隆及其在叶片花青素合成中的功能[J].生物技术通报,2026,62(1):82-98.
[32] 邓美欣,吴奇轩,龙健梅,等.黄梁木逆境响应因子NcMYC2基因的生物学功能分析[J].植物生理学报,2026,62(1):82-98.
[33] OHNUMA T, NAKAYAMA S, ANAN E, et al.Activation of the Nrf2/ARE pathway via S-alkylation of cysteine 151 in the chemopreventive agent-sensor Keap1 protein by falcarindiol, a conjugated diacetylene compound[J]. Toxicology and applied pharmacology, 2010, 244(1): 27-36.
[34] WANG Y Z, ZHANG Y C, CHENG J S, et al.Protective effects of BML-111 on cerulein-induced acute pancreatitis-associated lung injury via activation of Nrf2/ARE signaling pathway[J]. Inflammation, 2014, 37(4): 1120-1133.
[35] 廖霞,郑少杰,卢可可,等.植物多酚通过Nrf2/ARE信号通路抗氧化作用研究进展[J].食品科学,2016,37(7):227-232.
[36] ZHOU Y F, LAN Q Q, YU W H, et al.Analysis of the small auxin-up RNA (SAUR) genes regulating root growth angle (RGA) in apple[J]. Genes, 2022, 13(11):2121.
[37] PENG Z, LI W X, GAN X Q, et al.Genome-wide analysis of SAUR gene family identifies a candidate associated with fruit size in loquat (Eriobotrya japonica Lindl.)[J]. International journal of molecular sciences, 2022, 23(21): 13271.
[38] 贾秀苹,王莹,卯旭辉,等.盐碱胁迫对植物的影响及抗性机制研究进展[J].寒旱农业科学,2024,3(7):593-599.
[39] LIU J, WANG Y Q, LI Q T.Analysis of differentially expressed genes and adaptive mechanisms of Prunus triloba Lindl. under alkaline stress[J]. Hereditas, 2017, 154(1): 10.
[40] 李源,刘贵波,高洪文,等.紫花苜蓿种质耐盐性综合评价及盐胁迫下的生理反应[J].草业学报,2010,19(4):79-86.
[41] 陈广琳,陈友媛,孙萍,等.滨海区黄花鸢尾的耐盐碱性及氮磷去除效果[J].中国环境科学,2015,35(11):3422-3430.
[42] 李善家,韩多红,王恩军,等.外源甜菜碱对盐胁迫下黑果枸杞种子萌发和幼苗保护酶活性的影响[J].草业科学,2016,33(4):674-680.
[43] 米永伟,陈垣,郭凤霞,等.盐胁迫下黑果枸杞幼苗对外源甜菜碱的生理响应[J].草业科学,2012,29(9):1417-1421.