Abstract
Plants are sessile organisms, which are vulnerable to environmental stresses. As such, plants have developed multiple molecular, physiological, and cellular mechanisms to cope with natural stressors. However, these environmental adversities, including drought, are sources of the main agribusiness problems since they interfere with plant growth and productivity. Particularly under water deprivation conditions, the abscisic acid-responsive element-binding protein AREB1/ABF2 plays an important role in drought stress response and physiological adaptation. In this investigation, we provide substantial confirmation for the role of AREB1/ABF2 in plant survival under severe water deficit using the CRISPR activation (CRISPRa) technique to enhance the AREB1 gene expression. In our strategy, the inactive nuclease dCas9 was fused with an Arabidopsis histone acetyltransferase 1, which improves gene expression by remodeling chromatin. The AREB1 overexpression promotes an improvement in the physiological performance of the transgenic homozygous plants under drought, which was associated with an increase in chlorophyll content, antioxidant enzyme activity, and soluble sugar accumulation, leading to lower reactive oxygen species accumulation. Finally, we found that the CRISPR-mediated up-regulation of AREB1 changes the abundance of several downstream ABA-inducible genes, allowing us to report that CRISPRa dCas9-HAT is a valuable biotechnological tool to improve drought stress tolerance through the positive regulation of AREB1.
Highlights
Plants are sessile organisms, which are vulnerable to environmental stresses
The transcriptional modulation of AREB-1 expression by CRISPR was capable of upregulating many important genes related to drought adaptability through the abscisic acid (ABA)-dependent signaling pathway in plants
Our study shows that overexpression of AREB-1 mediated by the CRISPR activation (CRISPRa) strategy results in highly improved drought stress responses, encompassing a powerful biotechnology tool for genetic engineering
Summary
Plants are sessile organisms, which are vulnerable to environmental stresses. As such, plants have developed multiple molecular, physiological, and cellular mechanisms to cope with natural stressors. Under water deprivation conditions, the abscisic acid-responsive element-binding protein AREB1/ABF2 plays an important role in drought stress response and physiological adaptation In this investigation, we provide substantial confirmation for the role of AREB1/ABF2 in plant survival under severe water deficit using the CRISPR activation (CRISPRa) technique to enhance the AREB1 gene expression. ROS can act as signaling molecules that regulates several stress-associated processes, encompassing other protective mechanisms, such as the production and accumulation of osmolytes, electron carriers and improvement of transcription, translation, and activity of antioxidant enzymes, among others[1,2,3] These mechanisms comprise a sophisticated and intricate hormone-responsive pathway, whose transcription factors appear as nodes on signal integration and gene expression remodeling. The overexpression of AREB-1 under the CaMV 35S constitutive promoter has been demonstrated to be a good strategy for drought tolerance improvement in different plant species, such as Arabidopsis, rice, and s oybean[14,15,16,17]
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