PtrMYB3, a R2R3-MYB Transcription Factor from Poncirus trifoliata, Negatively Regulates Salt Tolerance and Hydrogen Peroxide Scavenging

Tonglu Wei,Dalong Guo,Jihong Liu

doi:10.3390/antiox10091388

Tonglu Wei, Dalong Guo + Show 1 more

Open Access

https://doi.org/10.3390/antiox10091388

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Abstract

MYB transcription factors are widely present in plants and play significant roles in abiotic stresses. However, most MYB genes have not been identified in plants and their functions in abiotic stresses are still unknown. In this study, one MYB gene, designated as PtrMYB3, was cloned from trifoliate orange (Poncirus trifoliata (L.) Raf.), and its function in salt tolerance was investigated. PtrMYB3 contains a conserved R2R3-MYB domain, which is the typical property of R2R3-MYB subfamily proteins. Expression profiling under abiotic stresses indicated that PtrMYB3 could be induced by salt, dehydration and cold stresses. PtrMYB3 was found to be localized to the nucleus and possessed transactivation activity. Overexpression of PtrMYB3 by genetic transformation in tobacco impaired its salt tolerance, whereas silencing of PtrMYB3 by VIGS (virus-induced gene silencing) in trifoliate orange conferred significantly enhanced salt tolerance, indicating that PtrMYB3 negatively regulates salt tolerance. Furthermore, a peroxidase gene (PtrPOD) was found to be greatly upregulated in PtrMYB3-silenced trifoliate orange, and a dual LUC (luciferase) assay confirmed that PtrMYB3 could suppress the expression of PtrPOD. The hydrogen peroxide (H2O2) accumulation in PtrMYB3 transgenic tobacco plants after salt stress was higher than the wild type (WT), further confirming that overexpression of PtrMYB3 inhibited PtrPOD-mediated H2O2 scavenging. Taken together, these results demonstrate that PtrMYB3 negatively regulates salt tolerance, at least in part, due to the excess accumulation of H2O2.

Highlights

Salt stress is becoming a major environmental problem because of the massive crop loss caused by high salinity in soil, especially in arid and semi-arid regions around the world [1]
By BLAST analysis in the Arabidopsis genome database, the protein sequence of this gene showed the highest identity with AtMYB3 (AT1G22640), and one R2R3-MYB transcription factor
Cloning and sequence analysis showed that PtrMYB3 is a typical R2R3-MYB transcription factor

Summary

Introduction

Salt stress is becoming a major environmental problem because of the massive crop loss caused by high salinity in soil, especially in arid and semi-arid regions around the world [1]. Salt stress results in ion toxicity, osmotic stress and oxidative stress [2], leading to severe damage to plants. It is imperative to improve plant salt tolerances by employing various strategies. Genetic engineering is an effective approach for creating tolerant transgenic plants tolerant to salt stress, and one of the key steps is to identify important salt-responsive genes. To this end, it is pressing and important to identify key genes and unravel their functions in salt tolerance. Transcription factors are important regulatory proteins that can either activate or inhibit the expression of downstream target genes by binding directly to the promoters [3]

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