Abstract

Previously, we reported that overexpression of AtRH17, an Arabidopsis DEAD-box RNA helicase gene, confers salt stress-tolerance via a pathway other than the well-known salt stress-responsive pathways. To decipher the salt stress-responsive pathway in AtRH17-overexpressing transgenic plants (OXs), we performed RNA-Sequencing and identified 397 differentially expressed genes between wild type (WT) and AtRH17 OXs. Among them, 286 genes were upregulated and 111 genes were downregulated in AtRH17 OXs relative to WT. Gene ontology annotation enrichment and KEGG pathway analysis showed that the 397 upregulated and downregulated genes are involved in various biological functions including secretion, signaling, detoxification, metabolic pathways, catabolic pathways, and biosynthesis of secondary metabolites as well as in stress responses. Genevestigator analysis of the upregulated genes showed that nine genes, namely, LEA4-5, GSTF6, DIN2/BGLU30, TSPO, GSTF7, LEA18, HAI1, ABR, and LTI30, were upregulated in Arabidopsis under salt, osmotic, and drought stress conditions. In particular, the expression levels of LEA4-5, TSPO, and ABR were higher in AtRH17 OXs than in WT under salt stress condition. Taken together, our results suggest that a high AtRH17 expression confers salt stress-tolerance through a novel salt stress-responsive pathway involving nine genes, other than the well-known ABA-dependent and ABA-independent pathways.

Highlights

  • RNA helicases (RHs) are present in most prokaryotic and eukaryotic organisms, which catalyze the unwinding of DNA or the secondary structure of RNA, and play essential roles in almost every aspect of genetic processes such as replication, transcription, translation, repair, and recombination [1,2]

  • We reported that overexpression of AtRH17 confers salt stress-tolerance at the seedling and mature plant stages

  • The well-known ABA-dependent and ABA-independent salt stress-responsive genes such as RD29A, RAB18, RD29B, RD22, COR47, DREB2A, and DREB2B showed similar or lower expression levels in AtRH17 overexpressing transgenic plants (OXs) than in wild type (WT) under salt stress conditions [17], implying that salt stress-tolerance of AtRH17 OXs is mediated by an uncharacterized pathway or mechanism other than the well-known stress-responsive pathways

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Summary

Introduction

RNA helicases (RHs) are present in most prokaryotic and eukaryotic organisms, which catalyze the unwinding of DNA or the secondary structure of RNA, and play essential roles in almost every aspect of genetic processes such as replication, transcription, translation, repair, and recombination [1,2]. Multiple studies have suggested that DEAD-box RHs play essential roles in abiotic stress responses in plants through their functions in specific RNA processing events [10,11]. To elucidate the salt stress-responsive pathway in AtRH17 OXs, we performed RNA-Sequencing (RNA-Seq) and analyzed the expression of Arabidopsis genes in WT and AtRH17 OXs. RNA-Seq analysis showed 286 upregulated and 111 downregulated genes in AtRH17 OXs compared to WT. Further analysis of the upregulated genes suggested that overexpression of AtRH17 confers salt stress-tolerance via pathway(s) involving LEA4-5, GSTF6, DIN2/BGLU30, TSPO, GSTF7, LEA18, HAI1, ABR, and LTI30, other than the well-known ABA-dependent and ABA-independent stress-responsive pathways in Arabidopsis

Transcriptomic Profiling of AtRH17 OXs
Analysis of Up- and Downregulated Genes in AtRH17 OXs
Identification of the Salt Stress-Tolerance Pathway in AtRH17 OXs
Discussion
Plant Materials and Growth Conditions
Plant Stress Treatment for Quantitative RT-PCR
RNA Isolation and First Strand cDNA Synthesis
Quantitative RT-PCR
Library Preparation and RNA-Sequencing
RNA-Seq Data Analysis
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