Dataset Information

Transcriptional modulation of AREB-1 by CRISPRa improves plant physiological performance under severe water deficit.

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.

SUBMITTER: de Melo BP

PROVIDER: S-EPMC7530729 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Transcriptional modulation of AREB-1 by CRISPRa improves plant physiological performance under severe water deficit.

de Melo Bruno Paes BP Lourenço-Tessutti Isabela Tristan IT Paixão Joaquin Felipe Roca JFR Noriega Daniel David DD Silva Maria Cristina Mattar MCM de Almeida-Engler Janice J Fontes Elizabeth Pacheco Batista EPB Grossi-de-Sa Maria Fatima MF

Scientific reports 20201001 1

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 ...[more]

PMID: 33004844

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Project description:Although mulberry cultivars Wubu, Yu711, and 7307 display distinct anatomical, morphological, and agronomic characteristics under natural conditions, it remains unclear if they differ in drought tolerance. To address this question and elucidate the underlying regulatory mechanisms at the whole-plant level, 2-month old saplings of the three mulberry cultivars were exposed to progressive soil water deficit for 5 days. The physiological responses and transcriptional changes of PIPs in different plant tissues were analyzed. Drought stress led to reduced leaf relative water content (RWC) and tissue water contents, differentially expressed PIPs, decreased chlorophyll and starch, increased soluble sugars and free proline, and enhanced activities of antioxidant enzymes in all plant parts of the three cultivars. Concentrations of hydrogen peroxide (H2O2), superoxide anion (O2 •-), and malonaldehyde (MDA) were significantly declined in roots, stimulated in leaves but unaltered in wood and bark. In contrast, except the roots of 7307, soluble proteins were repressed in roots and leaves but induced in wood and bark of the three cultivars in response to progressive water deficit. These results revealed tissue-specific drought stress responses in mulberry. Comparing to cultivar Yu711 and 7307, Wubu showed generally slighter changes in leaf RWC and tissue water contents at day 2, corresponding well to the steady PIP transcript levels, foliar concentrations of chlorophyll, O2 •-, MDA, and free proline. At day 5, Wubu sustained higher tissue water contents in green tissues, displayed stronger responsiveness of PIP transcription, lower concentrations of soluble sugars and starch, lower foliar MDA, higher proline and soluble proteins, higher ROS accumulation and enhanced activities of several antioxidant enzymes. Our results indicate that whole-plant level responses of PIP transcription, osmoregulation through proline and soluble proteins and antioxidative protection are important mechanisms for mulberry to cope with drought stress. These traits play significant roles in conferring the relatively higher drought tolerance of cultivar Wubu and could be potentially useful for future mulberry improvement programmes.

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Dataset Information

Transcriptional modulation of AREB-1 by CRISPRa improves plant physiological performance under severe water deficit.

Publications

Transcriptional modulation of AREB-1 by CRISPRa improves plant physiological performance under severe water deficit.

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