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

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.

SUBMITTER: Wei T

PROVIDER: S-EPMC8466168 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Tartary buckwheat (Fagopyrum tataricum Gaertn.) is a coarse cereal with strongly abiotic resistance. The MYB family plays a regulatory role in plant growth, development, and responses to biotic and abiotic stresses. However, the characteristics and regulatory mechanisms of MYB transcription factors in Tartary buckwheat remain unclarified. Here, this study cloned the FtMYB22 gene from Tartary buckwheat, and investigated its involvement in responding to individual water deficit and salt stress in Arabidopsis. Sequence analysis highlighted that the N-termini of FtMYB22 contained two highly conserved SANT domains and one conserved domain from the SG20 subfamily. Nucleus-localized FtMYB22 did not have individual transcriptional activation activity. Water deficiency and salt stress induced the high expression of the GUS gene, which was driven by the promoter of FtMYB22. Yeast stress experiments showed that the overexpression of FtMYB22 significantly reduced the growth activity of transgenic yeast under water deficit or salt stress. Consistently, the overexpression of FtMYB22 reduced the salt and water deficit stress resistance of the transgenic plants. In addition, physiological parameters showed that transgenic plants had lower proline and antioxidant enzyme activity under stress conditions. Compared to the wild-type (WT), transgenic plants accumulated more malondialdehyde (MDA), H2O2, and O2−; they also showed higher ion permeability and water loss rates of detached leaves under stress treatments. Notably, FtMYB22 was involved in plant stress resistance through an ABA-dependent pathway. Under stress conditions, the expression of RD29A, RD29B, PP2CA, KIN1, COR15A, and other genes in response to plant stress in transgenic lines was significantly lower than that in the WT (p < 0.05). Furthermore, yeast two-hybrid assay showed that there was a significant interaction between FtMYB22 and the ABA receptor protein RCAR1/2, which functioned in the ABA signal pathway. Altogether, FtMYB22, as a negative regulator, inhibited a variety of physiological and biochemical reactions, affected gene expression and stomatal closure in transgenic plants through the ABA-dependent pathway, and reduced the tolerance of transgenic Arabidopsis to water deficiency and salt stress. Based on these fundamental verifications, further studies would shed light on the hormone signal response mechanism of FtMYB22.

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

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