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The DOF-Domain Transcription Factor ZmDOF36 Positively Regulates Starch Synthesis in Transgenic Maize.

ABSTRACT: Starch synthesis is a complex process that influences crop yield and grain quality in maize. Many key enzymes have been identified in starch biosynthesis; however, the regulatory mechanisms have not been fully elucidated. In this study, we identified a DOF family gene, ZmDOF36, through transcriptome sequencing analysis. Real-time PCR indicated that ZmDOF36 was highly expressed in maize endosperm, with lower expression in leaves and tassels. ZmDOF36 is a typical DOF transcription factor (TF) that is localized to the nucleus and possesses transcriptional activation activity, and its transactivation domain is located in the C-terminus (amino acids 227-351). Overexpression of ZmDOF36 can increase starch content and decrease the contents of soluble sugars and reducing sugars. In addition, abnormal starch structure in transgenic maize was also observed by scanning electron microscopy (SEM). Furthermore, the expression levels of starch synthesis-related genes were up-regulated in ZmDOF36-expressing transgenic maize. ZmDOF36 was also shown to bind directly to the promoters of six starch biosynthesis genes, ZmAGPS1a, ZmAGPL1, ZmGBSSI, ZmSSIIa, ZmISA1, and ZmISA3 in yeast one-hybrid assays. Transient expression assays showed that ZmDOF36 can activate the expression of ZmGBSSI and ZmISA1 in tobacco leaves. Collectively, the results presented here suggest that ZmDOF36 acts as an important regulatory factor in starch synthesis, and could be helpful in devising strategies for modulating starch production in maize endosperm.

SUBMITTER: Wu J

PROVIDER: S-EPMC6474321 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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The DOF-Domain Transcription Factor ZmDOF36 Positively Regulates Starch Synthesis in Transgenic Maize.

Wu Jiandong J Chen Long L Chen Mingchao M Zhou Wei W Dong Qing Q Jiang Haiyang H Cheng Beijiu B

Frontiers in plant science 20190412

Starch synthesis is a complex process that influences crop yield and grain quality in maize. Many key enzymes have been identified in starch biosynthesis; however, the regulatory mechanisms have not been fully elucidated. In this study, we identified a DOF family gene, ZmDOF36, through transcriptome sequencing analysis. Real-time PCR indicated that ZmDOF36 was highly expressed in maize endosperm, with lower expression in leaves and tassels. ZmDOF36 is a typical DOF transcrip ...[more]

PMID: 31031791

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Project description:RationalePostischemic angiogenesis is critical to limit the ischemic tissue damage and improve the blood flow recovery. The regulation and the underlying molecular mechanisms of postischemic angiogenesis are not fully unraveled. TFEB (transcription factor EB) is emerging as a master gene for autophagy and lysosome biogenesis. However, the role of TFEB in vascular disease is less understood.ObjectiveWe aimed to determine the role of endothelial TFEB in postischemic angiogenesis and its underlying molecular mechanism.Methods and resultsIn primary human endothelial cells (ECs), serum starvation induced TFEB nuclear translocation. VEGF (vascular endothelial growth factor) increased TFEB expression level and nuclear translocation. Utilizing genetically engineered EC-specific TFEB transgenic and KO (knockout) mice, we investigated the role of TFEB in postischemic angiogenesis in the mouse hindlimb ischemia model. We observed improved blood perfusion and increased capillary density in the EC-specific TFEB transgenic mice compared with the wild-type littermates. Furthermore, blood flow recovery was attenuated in EC-TFEB KO mice compared with control mice. In aortic ring cultures, the TFEB transgene significantly increased vessel sprouting, whereas TFEB deficiency impaired the vessel sprouting. In vitro, adenovirus-mediated TFEB overexpression promoted EC tube formation, migration, and survival, whereas siRNA-mediated TFEB knockdown had the opposite effect. Mechanistically, TFEB activated AMPK (AMP-activated protein kinase)-α signaling and upregulated autophagy. Through inactivation of AMPKα or inhibition of autophagy, we demonstrated that the AMPKα and autophagy are necessary for TFEB to regulate angiogenesis in ECs. Finally, the positive effect of TFEB on AMPKα activation and EC tube formation was mediated by TFEB-dependent transcriptional upregulation of MCOLN1 (mucolipin-1).ConclusionsIn summary, our data demonstrate that TFEB is a positive regulator of angiogenesis through activation of AMPKα and autophagy, suggesting that TFEB constitutes a novel molecular target for ischemic vascular disease.

Dataset Information

The DOF-Domain Transcription Factor ZmDOF36 Positively Regulates Starch Synthesis in Transgenic Maize.

Publications

The DOF-Domain Transcription Factor ZmDOF36 Positively Regulates Starch Synthesis in Transgenic Maize.

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