Abstract

BackgroundHydrogen peroxide (H2O2) has been known to function as a signalling molecule involved in the modulation of various physiological processes in plants. H2O2 has been shown to act as a promoter during adventitious root formation in hypocotyl cuttings. In this study, RNA-Seq was performed to reveal the molecular mechanisms underlying H2O2-induced adventitious rooting.ResultsRNA-Seq data revealed that H2O2 treatment greatly increased the numbers of clean reads and expressed genes and abundance of gene expression relative to the water treatment. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses indicated that a profound change in gene function occurred in the 6-h H2O2 treatment and that H2O2 mainly enhanced gene expression levels at the 6-h time point but reduced gene expression levels at the 24-h time point compared with the water treatment. In total, 4579 differentially expressed (2-fold change > 2) unigenes (DEGs), of which 78.3% were up-regulated and 21.7% were down-regulated; 3525 DEGs, of which 64.0% were up-regulated and 36.0% were down-regulated; and 7383 DEGs, of which 40.8% were up-regulated and 59.2% were down-regulated were selected in the 6-h, 24-h, and from 6- to 24-h treatments, respectively. The number of DEGs in the 6-h treatment was 29.9% higher than that in the 24-h treatment. The functions of the most highly regulated genes were associated with stress response, cell redox homeostasis and oxidative stress response, cell wall loosening and modification, metabolic processes, and transcription factors (TFs), as well as plant hormone signalling, including auxin, ethylene, cytokinin, gibberellin, and abscisic acid pathways. Notably, a large number of genes encoding for heat shock proteins (HSPs) and heat shock transcription factors (HSFs) were significantly up-regulated during H2O2 treatments. Furthermore, real-time quantitative PCR (qRT-PCR) results showed that, during H2O2 treatments, the expression levels of ARFs, IAAs, AUXs, NACs, RD22, AHKs, MYBs, PIN1, AUX15A, LBD29, LBD41, ADH1b, and QORL were significantly up-regulated at the 6- and/or 24-h time points. In contrast, PER1 and PER2 were significantly down-regulated by H2O2 treatment. These qRT-PCR results strongly correlated with the RNA-Seq data.ConclusionsUsing RNA-Seq and qRT-PCR techniques, we analysed the global changes in gene expression and functional profiling during H2O2-induced adventitious rooting in mung bean seedlings. These results strengthen the current understanding of H2O2-induced adventitious rooting and the molecular traits of H2O2 priming in plants.

Highlights

  • Hydrogen peroxide (H2O2) has been known to function as a signalling molecule involved in the modulation of various physiological processes in plants

  • The sequencing data were used for bioinformatics analysis with the data from the same batch samples treated with water for 6 h (Wat6) or 24 h (Wat24) that were reported in our previous study [29]

  • Data analysis showed that the number of clean reads and expressed genes were increased by 33.47% in HO6 and 2.98% in HO24 and by 9.21% (6142 genes) in HO6 and 7.12% (4602 genes) in HO24 compared with Wat6 and Wat24, respectively [29]; and the expressed gene number were increased by 4.71% and 2.0% compared with 6- and 24-h Indole-3-butyric acid (IBA) treatments, respectively [30]

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Summary

Introduction

Hydrogen peroxide (H2O2) has been known to function as a signalling molecule involved in the modulation of various physiological processes in plants. The process of adventitious root formation provides an ideal experimental system with which to study the important physiological and molecular events that occur during the tissue dedifferentiation and morphogenesis [3]. Many exogenous and endogenous factors that play important roles in adventitious rooting have been extensively studied using physiological and molecular methods. Auxin is known to play a critical role in inducing cell dedifferentiation and root primordia formation in cuttings. Other molecules, such as hydrogen peroxide (H2O2) and nitric oxide (NO) that produced by cells in response to various stresses, act as signalling molecules that are involved in modulating of the induction and initiation of adventitious root formation [4]. The molecular mechanisms by which H2O2 promotes adventitious root development remain elusive

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