Heterologous expression of AaLac1 gene in hairy roots and its role in secondary metabolism under PEG-induced osmotic stress condition in Artemisia annua L.

IF 3.4 3区 生物学 Q1 PLANT SCIENCES
Sabitri Kumari, Nidhi Rai, Sneha Singh, Pajeb Saha, Mansi Singh Bisen, Shashi Pandey-Rai
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引用次数: 0

Abstract

This study explores the Laccase gene (AaLac) family along with AaLac1 expression in hairy roots of A. annua. 42 AaLacs were identified by detecting three conserved domains: Cu-oxidase, Cu oxidase-2, and Cu oxidase-3. The physicochemical properties show that AaLacs are proteins with 541-1075 amino acids. These proteins are stable, with an instability index less than 40. Phylogenetic and motif studies have shown structural variants in AaLacs, suggesting functional divergence. 22 AaLac cis-regulatory elements were selected for their roles in drought stress, metabolic modulations, defense, and stress responses. A comparison of AtLac and AaLac proteins showed that 11 AtLacs mitigates stress reactions. In silico expression, analysis of 11 AtLacs showed that AtLac84 may function under osmotic stress. Thus, the Homolog AaLac1 was selected by expression profiling. The real-time PCR results showed that AaLac1 enhances osmotic stress tolerance in shoot and root samples. It was also used to analyze AaLac1, ADS, and CYP71AV1 gene expression in hairy roots via induction. The transformed hairy roots exhibited a greater capacity for PEG-induced osmotic stress tolerance in contrast to the untransformed roots. The gene expression analysis also depicted a significant increment in expression of AaLac1, ADS, and CYP71AV1 genes to 3.8, 6.9, and 3.1 folds respectively. The transformed hairy roots exhibited a significant increase of 2.2 and 1.4 fold in flavonoid and phenolic content respectively. Also, lignin content and artemisinin content increased by 7.05 folds and 95.6% with respect to the control. Thus, transformed hairy roots of A. annua under PEG-induced osmotic stress demonstrate the involvement of the AaLac1 gene in stress responses, lignin biosynthesis, and secondary metabolism production.

Supplementary information: The online version contains supplementary material available at 10.1007/s12298-024-01516-8.

毛根中 AaLac1 基因的异源表达及其在 PEG 诱导的黄花蒿渗透胁迫条件下的次生代谢中的作用
本研究探讨了漆酶基因(AaLac)家族以及 AaLac1 在鹅掌楸毛根中的表达。通过检测三个保守结构域,确定了 42 个 AaLac:铜氧化酶、铜氧化酶-2 和铜氧化酶-3。理化性质表明,AaLacs 是含有 541-1075 个氨基酸的蛋白质。这些蛋白质很稳定,不稳定指数小于 40。系统发育和主题研究显示 AaLacs 存在结构变异,表明其存在功能分化。22 个 AaLac 顺式调节元件因其在干旱胁迫、代谢调节、防御和胁迫反应中的作用而被选中。对 AtLac 和 AaLac 蛋白的比较表明,11 个 AtLacs 可减轻应激反应。对11个AtLacs的硅表达分析表明,AtLac84可能在渗透胁迫下发挥作用。因此,通过表达谱分析筛选出了同源物 AaLac1。实时 PCR 结果表明,AaLac1 能增强芽和根样本对渗透胁迫的耐受性。此外,还通过诱导分析了毛根中 AaLac1、ADS 和 CYP71AV1 基因的表达。与未转化的根相比,转化的毛细根表现出更强的 PEG 诱导的渗透胁迫耐受能力。基因表达分析还显示,AaLac1、ADS 和 CYP71AV1 基因的表达量分别显著增加了 3.8、6.9 和 3.1 倍。转化后的毛细根中黄酮类化合物和酚类化合物含量分别显著增加了 2.2 倍和 1.4 倍。此外,木质素含量和青蒿素含量也比对照组分别增加了 7.05 倍和 95.6%。因此,在PEG诱导的渗透胁迫下,转化的A. annua毛根证明了AaLac1基因参与了胁迫响应、木质素生物合成和次生代谢的产生:在线版本包含补充材料,可查阅 10.1007/s12298-024-01516-8。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
7.10
自引率
0.00%
发文量
126
期刊介绍: Founded in 1995, Physiology and Molecular Biology of Plants (PMBP) is a peer reviewed monthly journal co-published by Springer Nature. It contains research and review articles, short communications, commentaries, book reviews etc., in all areas of functional plant biology including, but not limited to plant physiology, biochemistry, molecular genetics, molecular pathology, biophysics, cell and molecular biology, genetics, genomics and bioinformatics. Its integrated and interdisciplinary approach reflects the global growth trajectories in functional plant biology, attracting authors/editors/reviewers from over 98 countries.
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