Functional analysis of a putative novel transcription factor Arabidopsis MYB-like protein designated AtMYBL, which contains two predicted DNA-binding domains, was performed. The physiological role of the R-R-type MYB-like transcription factor has not been reported in any plant. Analyses of an AtMYBL promoter-β-glucuronidase (GUS) construct revealed substantial gene expression in old leaves and induction of GUS activity by ABA and salt stress. AtMYBL-overexpressing plants displayed a markedly enhanced leaf senescence phenotype. Moreover, the ectopic expression of the AtMYBL gene was very significantly influential in senescence parameters including Chl content, membrane ion leakage and the expression of senescence-related genes. Although the seed germination rate was improved under ABA and saline stress conditions in the AtMYBL-overexpressing plants, decreased salt tolerance was evident compared with the wild type and atmybl RNA interference lines during later seedling growth when exposed to long-term salt stress, indicating that AtMYBL protein is able to developmentally regulate stress sensitivity. Furthermore, AtMYBL protein activated the transcription of a reporter gene in yeast. Green fluorescent protein-tagged AtMYBL was localized in the nuclei of transgenic Arabidopsis cells. Taken together, these results suggest that AtMYBL functions in the leaf senescence process, with the abiotic stress response implicated as a putative potential transcription factor.
Summary
We identified and functionally characterized the AtSKIP gene (), an Arabidopsis homologue of SNW/SKIP, under abiotic stresses. Although the SNW/SKIP protein has been implicated as a critical transcription cofactor, its biological functions have yet to be reported in any plant.
Recently, we have isolated Salt‐tolerance genes (SATs) via the overexpression screening of yeast with a maize cDNA library. One of the selected genes (SAT2) appeared to confer elevated tolerance to salt. Maize SAT2 cDNA encodes a homologue of the human SNW/SKIP transcriptional coregulator.
Treatment with salt, mannitol and abscisic acid induced AtSKIP expression. Ectopic expression of the AtSKIP gene modulated the induction of salt tolerance, dehydration resistance and insensitivity towards abscisic acid under stress conditions. By contrast, atskip antisense lines displayed reduced tolerance to abiotic stresses during germination. Moreover, a decrease in AtSKIP expression resulted in an abnormal phenotype. We further determined that the AtSKIP protein activated the transcription of a reporter gene in yeast. Green fluorescent protein‐tagged AtSKIP was localized in the nuclei of both onion cells and transgenic Arabidopsis cells.
Taken together, these results suggest that AtSKIP functions as both a positive regulator and putative potential transcription factor in the abiotic stress signalling pathway.
This study examined the volatile terpenes produced by rice seedlings in response to oxidative stress induced by various abiotic factors. Solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) analyses revealed that when exposed to UV-B radiation, rice seedlings emitted a bouquet of monoterpene mixtures in a time-dependent manner. The mixtures comprised limonene, sabinene, myrcene, α-terpinene, β-ocimene, γ-terpinene, and α-terpinolene. Among them, (S)-limonene was the most abundant volatile, discriminated by chiral SPME-GC-MS. The volatile profiles collected from rice plants treated with both γ-irradiation and H2O2 were identical to those observed in the UV-B irradiated plants, thus indicating that the volatile mixtures were specifically produced in response to oxidative stress, particularly in the presence of H2O2. Using a reverse genetics approach, we isolated full-length rice terpene synthase 20 (OsTPS20, 599 amino acids, 69.39 kDa), which was further characterized as an (S)-limonene synthase by removing the N-terminal signal peptide (63 amino acids) of the protein. The recombinant OsTPS20 protein catalyzed the conversion of geranyl diphosphate to (S)-limonene and other minor monoterpenes, essentially covering all of the volatile compounds detected from the plant. Moreover, qRT-PCR revealed that the transcript levels of OsTPS20 were significantly induced in response to oxidative stress, thereby suggesting that OsTPS20 plays a major role in producing terpene volatiles during abiotic stress. Detailed biochemical analyses and the unusual domain characteristics of OsTPS20 are also discussed in this report.
The CCCH-type zinc finger proteins are a superfamily containing tandem zinc-binding motifs involved in many aspects of plant growth and development. However, the precise role of these proteins involved in plant stress tolerance is poorly understood. This study was to examine the regulatory and functional role of the CCCH-type zinc finger protein, AtOZF1 (At2g19810), under oxidative stress. Interestingly, the AtOZF1 protein was localized in the plasma membrane. The AtOZF1 transcripts were highly induced by treatment with hydrogen peroxide, abscisic acid and salinity. The AtOZF1-overexpressing plants were relatively resistant to oxidative stress than wild-type and T-DNA insertion mutant atozf1. Malondialdehyde, a decomposition product of lipid peroxidation, accumulated in atozf1 mutants more than in wild-type and AtOZF1-overexpressing plants. Furthermore, atozf1 mutants displayed lower activities of catalase and guaiacol peroxidase, higher chlorosis, and down-regulated expression of antioxidant genes under oxidative stress. Taken together, these observations demonstrate that AtOZF1 is required for the tolerance of Arabidopsis to oxidative stress.
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