• ISSN 1008-505X
  • CN 11-3996/S
肖燕, 姚珺玥, 廖琼, 吴秀文, 宋海星, 罗劲松, 张振华. 拟南芥生态型Tor-1根系响应镉的转录组学与可变剪接分析[J]. 植物营养与肥料学报, 2020, 26(8): 1513-1525. DOI: 10.11674/zwyf.19510
引用本文: 肖燕, 姚珺玥, 廖琼, 吴秀文, 宋海星, 罗劲松, 张振华. 拟南芥生态型Tor-1根系响应镉的转录组学与可变剪接分析[J]. 植物营养与肥料学报, 2020, 26(8): 1513-1525. DOI: 10.11674/zwyf.19510
XIAO Yan, YAO Jun-yue, LIAO Qiong, WU Xiu-wen, SONG Hai-xing, LUO Jin-song, ZHANG Zhen-hua. Transcriptome and alternative splicing analysis of cadmium response mechanisms in the root system of Arabidopsis thaliana ecotype Tor-1[J]. Journal of Plant Nutrition and Fertilizers, 2020, 26(8): 1513-1525. DOI: 10.11674/zwyf.19510
Citation: XIAO Yan, YAO Jun-yue, LIAO Qiong, WU Xiu-wen, SONG Hai-xing, LUO Jin-song, ZHANG Zhen-hua. Transcriptome and alternative splicing analysis of cadmium response mechanisms in the root system of Arabidopsis thaliana ecotype Tor-1[J]. Journal of Plant Nutrition and Fertilizers, 2020, 26(8): 1513-1525. DOI: 10.11674/zwyf.19510

拟南芥生态型Tor-1根系响应镉的转录组学与可变剪接分析

Transcriptome and alternative splicing analysis of cadmium response mechanisms in the root system of Arabidopsis thaliana ecotype Tor-1

  • 摘要:
    目的 研究拟南芥生态型根系对镉 (Cd) 的适应性机制,挖掘耐Cd基因,为培育修复型植物提供生理基础和理论指导。
    方法 本研究以高Cd积累同时高耐受性的拟南芥生态型Tor-1为试验材料,测定Cd处理后根系的生理变化、氧化应激反应、抗氧化酶活性,结合转录组学和可变剪接分析,以期从分子水平上解析Tor-1根系对Cd的适应性机制。
    结果 Cd处理后,拟南芥生态型Tor-1的主根长度和根尖数没有明显差异,但根系总体积、总表面积显著降低,总根长极显著下降,表明根系受到损伤;根系丙二醛浓度较对照有所增加,但变化不显著;脯氨酸浓度显著下降;超氧化物歧化酶 (SOD) 和过氧化氢酶 (CAT) 活性显著增加。基因本位论 (GO) 富集分析显示,在生物过程中,差异表达基因 (DEGs) 在代谢过程和对化学物质的反应功能富集最为显著;在细胞组分中,DEGs在细胞外组分功能显著富集;在核酸功能中,DEGs在氧化应激活性和血红素结合功能显著富集。京都基因和基因组百科全书 (KEGG) 通路表明,高比例诱导的DEGs富集于7条KEGG途径,分别为苯丙素生物合成 (4.49%)、次生代谢物的生物合成 (14.45%)、代谢途径 (20.57%)、硫代葡萄糖苷 (GS) 生物合成 (0.89%)、谷胱甘肽 (GSH) 代谢 (2.04%)、苯丙氨酸和酪氨酸及色氨酸的生物合成 (1.39%)、苯丙氨酸代谢 (1.14%)。在Cd胁迫下,Tor-1根系苯丙素合成与代谢和GSH代谢相关基因表达大部分显著上调,而GS合成相关基因表达受到抑制,并发生可变剪接事件,其中内含子保留事件发生最多。
    结论 Tor-1根系在应对Cd胁迫时虽然根系形态受到一定损伤,但可通过增加抗氧化酶 (SOD、CAT) 活性减轻镉胁迫产生的危害,并积极通过调节苯丙素合成与代谢、GSH代谢以及抑制GS合成对应的基因来减轻Cd对植物的伤害,还可通过可变剪接来积极适应Cd胁迫。

     

    Abstract:
    Objectives The Arabidopsis thaliana ecotype was used to study the adaptive mechanism of root system to cadmium (Cd) stress, providing physiological basis and theoretical guidance for the development of Cd resistance gene and the cultivation of phytoremediation materials.
    Methods The Arabidopsis thaliana ecotype Tor-1, with high Cd accumulation and high tolerance to Cd, was used to study the adaptive molecular mechanism of Tor-1 roots to Cd by analyzing the physiological changes, oxidative stress response and antioxidant enzyme activities, combining the transcriptome and alternative splicing analysis.
    Results Under Cd treatment, there was no significant difference in the length of main root and number of root tips in Tor-1, while the total root volume and surface area decreased, and the total root length decreased significantly, which all indicated that the roots of Tor-1 were damaged. Compared with the control, the malondialdehyde (MDA) concentration was slightly increased while proline concentration decreased significantly, and superoxide dismutase (SOD) and catalase (CAT) increased significantly under Cd treatment. GO enrichment showed that differential expression genes (DEGs) most significantly enriched in “the metabolic process” and “response to chemical” in the biological process category; in the cell components category, DEGs most significantly enriched in “the extracellular component function”; in the molecular function category, DEGs most significantly enriched in “antioxidant activity” and “heme binding function”. The Kyoto encyclopedia of genes and genomes (KEGG) pathway showed that a high proportion of induced DEGs was enriched in seven KEGG pathways, which were phenylpropanoid biosynthesis (4.49%); biosynthesis of secondary metabolites (14.45%); metabolic pathway (20.57%); glucosinolate biosynthesis (0.89%); glutathione metabolism (2.04%); henylalanine, tyrosine and tryptophan biosynthesis (1.39%); and phenylalanine metabolism (1.14%). Under Cd treatment, the expression of most genes, which were related to phenylpropanoid biosynthesis and metabolism and glutathione (GSH) metabolism, were significantly up-regulated, while the expression of genes related to glucoside (GS) synthesis was inhibited, and alternative splicing was occurred simultaneously in the roots, in which the intron retention events occurred frequently.
    Conclusions Although the root morphology of Tor-1 will be damaged under Cd stress, it could still increase antioxidant enzymes (SOD, CAT), and actively reduce the damage of Cd to plants by regulating the biosynthesis and metabolism of phenylpropanoid, glucosinolate biosynthesis and glutathione metabolism, and through alternative splicing to actively adapt to Cd stress.

     

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