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

doi:10.14088/j.cnki.issn0439-8114.2024.10.030

Abstract

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

CLC Number:

S642.2

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.

References

[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.