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基于 iTRAQ 的蛋白质组学筛选研究小麦(Triticum aestivum L.)根系细胞壁组分对铝胁迫响应的潜在调控因子。

iTRAQ-based proteomics screen for potential regulators of wheat (Triticum aestivum L.) root cell wall component response to Al stress.

机构信息

Key Laboratory of Sustainable Development and Utilization of Famous-region Drug, Key Laboratory of Panax Notoginseng Resources Sustainable Development and Utilization of State Administration of Traditional Chinese Medicine, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, Yunnan 650500, China.

Kunming Medical University, Kunming, Yunnan 650500, China.

出版信息

Gene. 2018 Oct 30;675:301-311. doi: 10.1016/j.gene.2018.07.008. Epub 2018 Jul 3.

Abstract

OBJECTIVE

The inhibition of Aluminum (Al)-induced root tip cell elongation is a major cause of plant root elongation suppression. The inhibition of root tip cell elongation is caused by a disruption of cell wall component metabolism, growth signaling, or cellular damage. The aim of this study was to identify the proteins involved in the metabolism of the root cell wall components under Al stress in the Al-tolerant wheat (Triticum aestivum L.) cultivar ET8.

METHODS

Differentially expressed proteins of Al-tolerant wheat roots were screened via isobaric tags for relative and absolute quantification (iTRAQ). Furthermore, their expression changes were detected via RT-PCR analysis. The contents of major components of the cell wall and their changes in metabolic enzyme activities were also investigated.

RESULTS

A total of 97 differentially expressed proteins from Al-tolerant wheat roots were screened and nine of these 97 proteins were root cell wall component related. The known nucleic acid sequences of proteins were 14-3-3 protein, the plasm membrane (PM) H-ATPase, phospholipase D, peroxidase, and glycosyltransferase. For 14-3-3 protein, phospholipase D and peroxidase, the protein expression and mRNA expression were consistent with Al-treatment; however, for PM H-ATPase and glycosyltransferase, the protein expression and mRNA expression were inconsistent under Al-stress. Furthermore, both cellulase activity and callase activity were down-regulated by Al stress, while the phenylalanineammonialyase (PAL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) activities were up-regulated. Furthermore, the PM H-ATPase activity was decreased in response to Al stress. In addition, the contents of callose, cellulose, lignin, and HO varied significantly.

CONCLUSIONS

The cell wall components, relative metabolism enzymes activity, and gene expression also changed followed by protein expression changed in response to Al stress. The results suggest that Al stress leads to marked variations in metabolic enzyme activity, carbohydrate content, followed by changes of root cell components in wheat roots.

摘要

目的

抑制铝(Al)诱导的根尖细胞伸长是植物根尖伸长抑制的主要原因。根尖细胞伸长的抑制是由于细胞壁成分代谢、生长信号或细胞损伤的破坏。本研究的目的是鉴定在耐铝小麦(Triticum aestivum L.)品种 ET8 中,Al 胁迫下根细胞壁成分代谢相关的蛋白质。

方法

通过同位素相对和绝对定量(iTRAQ)筛选耐铝小麦根系差异表达蛋白。此外,通过 RT-PCR 分析检测其表达变化。还研究了细胞壁主要成分的含量及其代谢酶活性的变化。

结果

从耐铝小麦根中筛选出 97 种差异表达蛋白,其中 9 种与根细胞壁成分有关。已知蛋白质的核酸序列为 14-3-3 蛋白、质膜(PM)H+-ATP 酶、磷脂酶 D、过氧化物酶和糖基转移酶。对于 14-3-3 蛋白、磷脂酶 D 和过氧化物酶,蛋白表达和 mRNA 表达与 Al 处理一致;然而,对于 PM H+-ATP 酶和糖基转移酶,Al 胁迫下蛋白表达和 mRNA 表达不一致。此外,纤维素酶活性和果胶酶活性均受 Al 胁迫下调,而苯丙氨酸氨裂解酶(PAL)、肉桂醇脱氢酶(CAD)和过氧化物酶(POD)活性上调。此外,PM H+-ATP 酶活性随 Al 胁迫而降低。此外,几丁质、纤维素、木质素和 HO 的含量也有显著变化。

结论

细胞壁成分、相对代谢酶活性和基因表达也随 Al 胁迫而变化,随后蛋白表达发生变化。结果表明,Al 胁迫导致小麦根系代谢酶活性、碳水化合物含量显著变化,进而导致根细胞成分发生变化。

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