拟南芥中异源过表达CsLOX6和CsHPL2基因提高植株耐盐性和耐旱性

doi:10.14088/j.cnki.issn0439-8114.2024.10.030

湖北农业科学 ›› 2024, Vol. 63 ›› Issue (10): 165-175.doi: 10.14088/j.cnki.issn0439-8114.2024.10.030

拟南芥中异源过表达CsLOX6和CsHPL2基因提高植株耐盐性和耐旱性

王聪, 李强, 柴琳, 王恒, 余宏军, 蒋卫杰

中国农业科学院蔬菜花卉研究所/蔬菜生物育种全国重点实验室,北京 100081

收稿日期:2023-11-05 出版日期:2024-10-25 发布日期:2024-11-05
通讯作者: 蒋卫杰（1965-）,男,北京人,研究员,主要从事蔬菜栽培生理和无土栽培研究,（电子信箱）jiangweijie@caas.cn。
作者简介:王聪（1995-）,女,河北石家庄人,硕士,主要从事蔬菜栽培与生理研究,（电话）18803219590（电子信箱）wangcong333333@163.com。
基金资助:
国家大宗蔬菜产业技术体系项目（CARS-23-B07）; 国家自然科学基金项目（31672203）; 国家重点研发计划支持项目（2023YFD2300703）; 现代农业产业技术体系北京市创新团队项目（BAIC-2024）

Heterologous overexpression of CsLOX6 and CsHPL2 genes in Arabidopsis thaliana enhances plant salt and drought tolerance

WANG Cong, LI Qiang, CHAI Lin, WANG Heng, YU Hong-jun, JIANG Wei-jie

Institute of Vegetables, Flowers/State Key Laboratory of Vegetable Biobreeding, Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received:2023-11-05 Published:2024-10-25 Online:2024-11-05

摘要/Abstract

摘要： 探究CsLOX6基因和CsHPL2基因对拟南芥（Arabidopsis thaliana）生长以及对干旱和盐胁迫抗性的影响,以野生型拟南芥（WT）、转基因型拟南芥（OE-CsLOX6、OE-CsHPL2）、突变体型拟南芥（TB-Atlox5）为材料,通过表型观察、转录组和代谢组学分析等方法评估4种拟南芥的耐旱性、耐盐性。结果表明,OE-CsLOX6的地上部鲜重和根长均显著高于其他基因型植株,地下部鲜重明显高于其他基因型植株;OE-CsHPL2的地上部鲜重、地下部鲜重和根长明显低于其他基因型植株。OE-CsLOX6、OE-CsHPL2和WT的代谢物含量存在较大差异,OE-CsLOX6和WT有64个差异代谢物,其中有6个上调,58个下调;OE-CsHPL2和WT有63个差异代谢物,其中有9个上调,54个下调。过表达CsLOX6基因和CsHPL2基因可以提高拟南芥对盐胁迫和干旱胁迫的抗性。在盐胁迫和干旱胁迫下,OE-CsLOX6、OE-CsHPL2的丙二醛含量要显著低于野生型植株,膜质过氧化程度低,SOD、POD和CAT活性要明显高于野生型,提高了植株的抗氧化能力,叶片长势明显优于野生型植株。

关键词: 拟南芥（Arabidopsis thaliana）, 异源过表达, CsLOX6基因, CsHPL2基因, 耐盐性, 耐旱性, 胁迫

Abstract: To explore the effects of CsLOX6 gene and CsHPL2 gene on the growth of Arabidopsis thaliana and its resistance to drought and salt stress, wild-type Arabidopsis thaliana （WT）, transgenic Arabidopsis thaliana （OE-CsLOX6, OE-CsHPL2）, and mutant Arabidopsis thaliana （TB-Atlox5） were used as materials, and the drought and salt tolerance of the four Arabidopsis thaliana species were evaluated through phenotype observation, transcriptome, and metabolomics analysis. The results showed that the aboveground fresh weight and root length of OE-CsLOX6 were significantly higher than those of other genotypes, and the underground fresh weight was significantly higher than that of other genotypes; the aboveground fresh weight, underground fresh weight, and root length of OE-CsHPL2 plants were significantly lower than those of other genotypes. There were significant differences in the metabolite content between OE-CsLOX6, OE-CsHPL2, and WT. OE-CsLOX6 and WT had 64 differential metabolites, of which 6 were upregulated and 58 were downregulated; OE-CsHPL2 and WT had 63 differential metabolites, of which 9 were upregulated and 54 were downregulated. Overexpression of CsLOX6 gene and CsHPL2 gene could enhance the resistance of Arabidopsis thaliana to salt stress and drought stress. Under salt stress and drought stress, compared with wild-type plants, the malondialdehyde content of OE-CsLOX6 and OE-CsHPL2 was significantly lower, and the degree of membrane peroxidation was lower,the activities of SOD, POD, and CAT enzymes were significantly higher than those of the wild type, which improved the antioxidant capacity of the plants and resulted in significantly better leaf growth compared to the wild type plants.

Key words: Arabidopsis thaliana, heterologous overexpression, CsLOX6 gene, CsHPL2 gene, salt tolerance, drought tolerance, stress

中图分类号:

S642.2

王聪, 李强, 柴琳, 王恒, 余宏军, 蒋卫杰. 拟南芥中异源过表达CsLOX6和CsHPL2基因提高植株耐盐性和耐旱性[J]. 湖北农业科学, 2024, 63(10): 165-175.

WANG Cong, LI Qiang, CHAI Lin, WANG Heng, YU Hong-jun, JIANG Wei-jie. Heterologous overexpression of CsLOX6 and CsHPL2 genes in Arabidopsis thaliana enhances plant salt and drought tolerance[J]. HUBEI AGRICULTURAL SCIENCES, 2024, 63(10): 165-175.

参考文献

[1] LIGOR T, BUSZEWSKI B.Single-drop microextraction and gas chromatography-mass spectrometry for the determination of volatile aldehydes in fresh cucumbers[J]. Analytical and bioanalytical chemistry, 2008, 391(6): 2283-2289.
[2] DUDAREVA N, MURFITT L M, MANN C J, et al.Developmental regulation of methyl benzoate biosynthesis and emission in snapdragon flowers[J]. Plant cell, 2000, 12(6): 949-961.
[3] SCALA A,ALLMANN S,MIRABELLA R,et al.Green leaf volatiles:A plant’s multifunctional weapon against herbivores and pathogens[J]. International journal of molecular sciences, 2013, 14(9): 17781-17811.
[4] THANH H T, VERGOIGNAN C, CACHON R, et al.Recombinant hydroperoxide lyase for the production of aroma compounds: Effect of substrate on the yeast yarrowia lipolytica[J]. Journal of molecular catalysis enzymatic, 2008, 52(3): 146-152.
[5] HADI MA, ZHANG F J, WU F F, et al.Advances in fruit aroma volatile research[J]. Molecules, 2013, 18:8200-8229.
[6] MATSUI K, MIYAHARA C, WILKINSON J, et al.Fatty acid hydroperoxide lyase in tomato fruits: Cloning and properties of a recombinant enzyme expressed in Escherichia coli[J]. Bioscience biotechnology and biochemistry, 2000, 64(6): 1189-1196.
[7] HUANG S W,ZHANG Z H,GU X F,et al.The genome of the cucumber, Cucumis sativus L[J]. Nature genetics,2009,41:1275-1281.
[8] YAMAUCHI Y, KUNISHIMA M, MIZUTANI M, et al.Reactive short-chain leaf volatiles act as powerful inducers of abiotic stress-related gene expression[J]. Scientific reports, 2015, 5(1):8030.
[9] CHAUVIN A, CALDELARI D, WOLFENDER J, et al.Four 13-lipoxygenases contribute to rapid jasmonate synthesis in wounded Arabidopsis thaliana leaves: A role for lipoxygenase 6 in responses to long-distance wound signals[J]. New phytologist,2013,197(2): 566-575.
[10] 丁辉. 拟南芥At LOX3基因在非生物胁迫中的功能研究[D]. 广州:华南师范大学, 2014.
[11] FARAHANI A S,TAGHAVI S M.Profiling expression of lipoxygenase in cucumber during compatible and incompatible plant-pathogen interactions[J]. Physiology and molecular biology of plants, 2016, 22(1):175-177.
[12] CHEN G, HACKETT R, WALKER D, et al.Identification of a specific isoform of tomato lipoxygenase (TomloxC) involved in the generation of fatty acid-derived flavor compounds[J]. Plant physiology, 2004, 136(1): 2641-2651.
[13] HOU Y, MENG K, HAN Y, et al.The persimmon 9-lipoxygenase gene DkLOX3 plays positive roles in both promoting senescence and enhancing tolerance to abiotic stress[J]. Frontiers in plant science, 2015, 6:1073.
[14] MATSUI K, SHIBUTANI M, HASE T, et al.Bell pepper fruit fatty acid hydroperoxide lyase is a cytochrome P450 (CYP74B)[J]. Febs letters, 1996, 394(1):1-24.
[15] 赵凌,沈文飚, 翟虎渠, 等. 植物的脂氢过氧化物裂解酶[J]. 植物生理学通报, 2004(2):135-140.
[16] 刘苗苗,张然然,魏文霞,等.黄瓜脂氢过氧化物裂解酶基因13-CsHPL的克隆及其表达分析[J]. 分子植物育种,2018, 16(21):6907-6914.
[17] HALITSCHKE R, ZIEGLER J, KEINANEN M, et al.Silencing of hydroperoxide lyase and allene oxide synthase reveals substrate and defense signaling crosstalk in Nicotiana attenuata[J]. Plant journal, 2004, 40: 35-46.
[18] VANCANNEYT G, SANZ C, FARMAKI T, et al.Hydroperoxide lyase depletion in transgenic potato plants leads to an increase in aphid performance[J]. Proceedings of the national academy of sciences, 2001, 98(14):8139-8144.
[19] 赵世杰, 苍晶. 植物生理学实验指导[M]. 北京:中国农业出版社, 2016
[20] CATOLA S, MARINO G, EMILIANI G, et al.Physiological and metabolomic analysis of Punica granatum (L.) under drought stress[J]. Planta, 2016, 243(2): 441-449.
[21] 石如玲, 姜玲玲. 过氧化物酶体脂肪酸β氧化[J]. 中国生物化学与分子生物学报, 2009, 25(1):12-16.
[22] 罗晓峰, 戚颖, 孟永杰, 等. Karrikins信号传导通路及功能研究进展[J]. 遗传, 2016, 38(1): 52-61.
[23] HU T, ZENG H, HU Z, et al.Overexpression of the tomato 13-lipoxygenase gene TomloxD increases generation of endogenous jasmonic acid and resistance to cladosporium fulvumand high temperature[J]. Plant molecular biology reporter,2013,31(5):1141-1149.
[24] 陈璐, 刘博, 安炎黄,等. 干旱胁迫下磷脂酶Dδ和9-脂氧合酶对拟南芥茉莉酸合成及种子萌发的影响[J]. 生态学杂志, 2018, 37(9):2627-2636.
[25] WOO L C,SANG WOOK H,SUN H I,et al.The pepper lipoxygenase CaLOX1 plays a role in osmotic,drought and high salinity stress response[J]. Plant and cell physiology,2015,56(5):930-942.
[26] 郭凤芝,许颖妍,熊青,等. 玳玳花瓣脂氢过氧化物裂解酶基因cDNA的克隆及原核表达[J]. 热带作物学报, 2017,38(4):673-681.
[27] 谢鑫鑫,吴卫东,林碧英,等. 脂氢过氧化物裂解酶基因对生菜遗传转化的研究[J]. 热带作物学报, 2016, 37(5):856-861.

拟南芥中异源过表达CsLOX6和CsHPL2基因提高植株耐盐性和耐旱性

Heterologous overexpression of CsLOX6 and CsHPL2 genes in Arabidopsis thaliana enhances plant salt and drought tolerance

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	陈映僮, 张春霞, 姚富成, 肖泽恒, 章家恩. 福寿螺与本地螺种在干旱胁迫下的存活率与生理响应差异[J]. 湖北农业科学, 2024, 63(6): 86-93.
[2]	张奥深, 徐敏, 陈炼, 熊伟, 程长松, 汪红武. 干旱胁迫下苎麻生理指标和抗逆相关基因表达的变化[J]. 湖北农业科学, 2024, 63(5): 6-11.
[3]	王建伟, 贺晓岚, 王根平, 欧倩, 肖云娟, 杨雪丽. 黑麦草种子及幼苗对镉胁迫的生理响应[J]. 湖北农业科学, 2024, 63(5): 126-135.
[4]	乐有章, 王建军, 戢小梅, 迟文超, 林之希. 美味猕猴桃MYB家族鉴定及其对水淹胁迫的响应[J]. 湖北农业科学, 2024, 63(5): 215-222.
[5]	唐佳乐, 段海波, 王静. 干旱胁迫下氮素对猴樟幼苗生长及光合特性的影响[J]. 湖北农业科学, 2024, 63(4): 96-100.
[6]	刘彩婷, 叶颖婷, 范铭丰, 陈晓逸, 刘希华. 阿耶波多氏芽孢杆菌对盐胁迫下绿豆的促生效果[J]. 湖北农业科学, 2024, 63(10): 8-13.
[7]	缑一杰, 杨振华. 褪黑素预处理对草莓苗耐旱性的影响[J]. 湖北农业科学, 2023, 62(8): 95-101.
[8]	陈博, 范继红, 李玉舒, 邹原东, 石进朝, 于海群. 金野紫穗槐对干旱胁迫的响应[J]. 湖北农业科学, 2023, 62(7): 89-94.
[9]	毛可欣, 王海荣, 安淼, 吕巍, 李健, 李国田. 猕猴桃‘脐红'与‘徐香'越冬期转录组的比较分析[J]. 湖北农业科学, 2023, 62(5): 165-171.
[10]	王伟杰, 徐东川, 张秀如, 于明, 林慧彬. 环境胁迫对黄芩次生代谢产物的影响[J]. 湖北农业科学, 2023, 62(4): 114-117.
[11]	于华丽, 温四民, 张桂玲, 董传洁, 程乾. 重金属胁迫对向日葵种子萌发特征影响评价及花海品种筛选[J]. 湖北农业科学, 2023, 62(1): 122-127.
[12]	徐扬帆, 冯立辉, 陈文峰, 刘剑彤, 吴辰熙, 胡红娟. 三种湿地植物对盐胁迫响应的研究[J]. 湖北农业科学, 2022, 61(6): 83-87.
[13]	周建云, 余佳斌, 刁朝强, 覃青青, 王晓. 干旱胁迫下木醋液对烤烟光合特性及根系生长的影响[J]. 湖北农业科学, 2022, 61(4): 85-88.
[14]	刘芳, 张志刚, 方伟, 王开梅, 王月莹. 草地贪夜蛾热休克蛋白Hsp70的生物信息学分析[J]. 湖北农业科学, 2022, 61(24): 153-159.
[15]	谢艺, 倪艳林, 朱航, 朱叶青, 张艳, 邹华文, 张丙林. 不同耐/感玉米自交系对高温胁迫的响应及其恢复力[J]. 湖北农业科学, 2022, 61(22): 5-13.