A R2R3-MYB Transcription Factor Gene, BpMYB123, Regulates BpLEA14 to Improve Drought Tolerance in Betula platyphylla.
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ABSTRACT:
SUBMITTER: Lv K
PROVIDER: S-EPMC8702527 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:We isolated a R2R3-MYB transcription factor BpMYB106, which regulates photosynthesis in birch (Betula platyphylla Suk.). BpMYB106 mainly expresses in the leaf and shoot tip of birch, and its protein is localized in the nucleus. We further fused isolated a 1588 bp promoter of BpMYB106 and analyzed it by PLACE, which showed some cis-acting elements related to photosynthesis. BpMYB106 promoter β-glucuronidase (GUS) reporter fusion studies gene, the result, showed the GUS reporter gene in transgenic birch with BpMYB106 promoter showed strong activities in shoot tip, cotyledon margins, and mature leaf trichomes. The overexpression of BpMYB106 in birch resulted in significantly increased trichome density, net photosynthetic rate, and growth rate as compared with the wild-type birch. RNA-Seq profiling revealed the upregulation of several photosynthesis-related genes in the photosynthesis and oxidative phosphorylation pathways in the leaves of transgenic plants. Yeast one-hybrid analysis, coupled with transient assay in tobacco, revealed that BpMYB106 binds a MYB binding site MYB2 in differentially expressed gene promoters. Thus, BpMYB106 may directly activate the expression of a range of photosynthesis related genes through interacting with the MYB2 element in their promoters. Our study demonstrating the overexpression of BpMYB106-a R2R3-MYB transcription factor-upregulates the genes of the photosynthesis and oxidative phosphorylation pathways to improve photosynthesis.
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Project description:Plant MYB transcription factors control diverse biological processes, such as differentiation, development and abiotic stress responses. In this study, we characterized BplMYB46, an MYB gene from Betula platyphylla (birch) that is involved in both abiotic stress tolerance and secondary wall biosynthesis. BplMYB46 can act as a transcriptional activator in yeast and tobacco. We generated transgenic birch plants with overexpressing or silencing of BplMYB46 and subjected them to gain- or loss-of-function analysis. The results suggest that BplMYB46 improves salt and osmotic tolerance by affecting the expression of genes including SOD, POD and P5CS to increase both reactive oxygen species scavenging and proline levels. In addition, BplMYB46 appears to be involved in controlling stomatal aperture to reduce water loss. Overexpression of BplMYB46 increases lignin deposition, secondary cell wall thickness and the expression of genes in secondary cell wall formation. Further analysis indicated that BplMYB46 binds to MYBCORE and AC-box motifs and may directly activate the expression of genes involved in abiotic stress responses and secondary cell wall biosynthesis whose promoters contain these motifs. The transgenic BplMYB46-overexpressing birch plants, which have improved salt and osmotic stress tolerance, higher lignin and cellulose content and lower hemicellulose content than the control, have potential applications in the forestry industry.
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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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Project description: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.
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Project description:MYB transcription factors of the R2R3-MYB family have been shown to play important roles in many plant processes. A sugarcane R2R3-MYB gene (ScMYB2) and its two alternative forms of transcript (ScMYB2S1 and ScMYB2S2) were identified in this study. The deduced protein of ScMYB2S1 is a typical plant R2R3-MYB protein, while ScMYB2S2 encodes a truncated protein. Real-time qPCR analysis revealed that ScMYB2S1 is suppressed under PEG-simulated drought stress in sugarcane, while ScMYB2S2 is induced at later treatment stage. A senescence symptom was observed when ScMYB2S1 was injected into tobacco leaves mediated by Agrobacterium, but no symptom for ScMYB2S2. Further investigation showed that the expression levels of 4 senescence-associated genes, NtPR-1a, NtNYC1, NtCAT3 and NtABRE, were markedly induced in tobacco leaves after ScMYB2S1-injection, while they were not sensitive to ScMYB2S2-injection. Moreover, MDA and proline were also investigated after injection. Similarly, MDA and proline levels were induced by ABA and ScMYB2S1, while inhibited by ScMYB2S2. We propose that ScMYB2, by alternatively splicing two transcripts (ScMYB2S1 and ScMYB2S2), is involved in an ABA-mediated leaf senescence signaling pathway and play positive role in respond to drought-induced senescence in sugarcane. The results of this study provide information for further research in sugarcane stress processes.
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Project description:Isoflavones comprise a group of secondary metabolites produced almost exclusively by plants in the legume family, including soybean [Glycine max (L.) Merr.]. They play vital roles in plant defense and have many beneficial effects on human health. Isoflavone content is a complex quantitative trait controlled by multiple genes, and the genetic mechanisms underlying isoflavone biosynthesis remain largely unknown. Via a genome-wide association study (GWAS), we identified 28 single nucleotide polymorphisms (SNPs) that are significantly associated with isoflavone concentrations in soybean. One of these 28 SNPs was located in the 5'-untranslated region (5'-UTR) of an R2R3-type MYB transcription factor, GmMYB29, and this gene was thus selected as a candidate gene for further analyses. A subcellular localization study confirmed that GmMYB29 was located in the nucleus. Transient reporter gene assays demonstrated that GmMYB29 activated the IFS2 (isoflavone synthase 2) and CHS8 (chalcone synthase 8) gene promoters. Overexpression and RNAi-mediated silencing of GmMYB29 in soybean hairy roots resulted in increased and decreased isoflavone content, respectively. Moreover, a candidate-gene association analysis revealed that 11 natural GmMYB29 polymorphisms were significantly associated with isoflavone contents, and regulation of GmMYB29 expression could partially contribute to the observed phenotypic variation. Taken together, these results provide important genetic insights into the molecular mechanisms underlying isoflavone biosynthesis in soybean.
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Project description:Herba epimedii (Epimedium), a traditional Chinese medicine, has been widely used as a kidney tonic and antirheumatic medicine for thousands of years. The bioactive components in herba epimedii are mainly prenylated flavonol glycosides, end-products of the flavonoid pathway. Epimedium species are also used as garden plants due to the colorful flowers and leaves. Many R2R3-MYB transcription factors (TFs) have been identified to regulate the flavonoid and anthocyanin biosynthetic pathways. However, little is known about the R2R3-MYB TFs involved in regulation of the flavonoid pathway in Epimedium. Here, we reported the isolation and functional characterization of the first R2R3-MYB TF (EsMYBA1) from Epimedium sagittatum (Sieb. Et Zucc.) Maxim. Conserved domains and phylogenetic analysis showed that EsMYBA1 belonged to the subgroup 6 clade (anthocyanin-related MYB clade) of R2R3-MYB family, which includes Arabidopsis AtPAP1, apple MdMYB10 and legume MtLAP1. EsMYBA1 was preferentially expressed in leaves, especially in red leaves that contain higher content of anthocyanin. Alternative splicing of EsMYBA1 resulted in three transcripts and two of them encoded a MYB-related protein. Yeast two-hybrid and transient luciferase expression assay showed that EsMYBA1 can interact with several bHLH regulators of the flavonoid pathway and activate the promoters of dihydroflavonol 4-reductase (DFR) and anthocyanidin synthase (ANS). In both transgenic tobacco and Arabidopsis, overexpression of EsMYBA1 induced strong anthocyanin accumulation in reproductive and/or vegetative tissues via up-regulation of the main flavonoid-related genes. Furthermore, transient expression of EsMYBA1 in E. sagittatum leaves by Agrobacterium infiltration also induced anthocyanin accumulation in the wounded area. This first functional characterization of R2R3-MYB TFs in Epimedium species will promote further studies of the flavonoid biosynthesis and regulation in medicinal plants.
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