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Increased Cuticle Waxes by Overexpression of WSD1 Improves Osmotic Stress Tolerance in Arabidopsis thaliana and Camelina sativa.

ABSTRACT: To ensure global food security under the changing climate, there is a strong need for developing 'climate resilient crops' that can thrive and produce better yields under extreme environmental conditions such as drought, salinity, and high temperature. To enhance plant productivity under the adverse conditions, we constitutively overexpressed a bifunctional wax synthase/acyl-CoA:diacylglycerol acyltransferase (WSD1) gene, which plays a critical role in wax ester synthesis in Arabidopsis stem and leaf tissues. The qRT-PCR analysis showed a strong upregulation of WSD1 transcripts by mannitol, NaCl, and abscisic acid (ABA) treatments, particularly in Arabidopsis thaliana shoots. Gas chromatography and electron microscopy analyses of Arabidopsis seedlings overexpressing WSD1 showed higher deposition of epicuticular wax crystals and increased leaf and stem wax loading in WSD1 transgenics compared to wildtype (WT) plants. WSD1 transgenics exhibited enhanced tolerance to ABA, mannitol, drought and salinity, which suggested new physiological roles for WSD1 in stress response aside from its wax synthase activity. Transgenic plants were able to recover from drought and salinity better than the WT plants. Furthermore, transgenics showed reduced cuticular transpirational rates and cuticle permeability, as well as less chlorophyll leaching than the WT. The knowledge from Arabidopsis was translated to the oilseed crop Camelina sativa (L.) Crantz. Similar to Arabidopsis, transgenic Camelina lines overexpressing WSD1 also showed enhanced tolerance to drought stress. Our results clearly show that the manipulation of cuticular waxes will be advantageous for enhancing plant productivity under a changing climate.

SUBMITTER: Abdullah HM

PROVIDER: S-EPMC8153268 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundTrehalose-6-phosphate phosphatases (TPPs), which are encoded by members of the TPP gene family, can improve the drought tolerance of plants. However, the molecular mechanisms underlying the dynamic regulation of TPP genes during drought stress remain unclear. In this study, we explored the function of an Arabidopsis TPP gene by conducting comparative analyses of a loss-of-function mutant and overexpression lines.ResultsThe loss-of-function mutation of Arabidopsis thaliana TPPF, a member of the TPP gene family, resulted in a drought-sensitive phenotype, while a line overexpressing TPPF showed significantly increased drought tolerance and trehalose accumulation. Compared with wild-type plants, tppf1 mutants accumulated more H2O2 under drought, while AtTPPF-overexpressing plants accumulated less H2O2 under drought. Overexpression of AtTPPF led to increased contents of trehalose, sucrose, and total soluble sugars under drought conditions; these compounds may play a role in scavenging reactive oxygen species. Yeast one-hybrid and luciferase activity assays revealed that DREB1A could bind to the DRE/CRT element within the AtTPPF promoter and activate the expression of AtTPPF. A transcriptome analysis of the TPPF-overexpressing plants revealed that the expression levels of drought-repressed genes involved in electron transport activity and cell wall modification were upregulated, while those of stress-related transcription factors related to water deprivation were downregulated. These results indicate that, as well as its involvement in regulating trehalose and soluble sugars, AtTPPF is involved in regulating the transcription of stress-responsive genes.ConclusionAtTPPF functions in regulating levels of trehalose, reactive oxygen species, and sucrose levels during drought stress, and the expression of AtTPPF is activated by DREB1A in Arabidopsis. These findings shed light on the molecular mechanism by which AtTPPF regulates the response to drought stress.

Project description:BackgroundCamelina sativa (L.) Crantz, known by such popular names as "gold-of-pleasure" and "false flax," is an alternative oilseed crop for biofuel production and can be grown in harsh environments. Considerable interest is now being given to the new concept of the development of a fusion plant which can be used as a soil remediation plant for ground contaminated by heavy metals as well as a bioenergy crop. However, knowledge of the transport processes for heavy metals across Camelina plant membranes is still rudimentary.ResultsFirstly, to investigate whether Camelina HMA (heavy metal P1B-ATPase) genes could be used in such a plant, we analyzed the expression patterns of eight HMA genes in Camelina (taken from the root, leaf, stem, flower, and silique). CsHMA3 genes were expressed in all organs. In addition, CsHMA3 was induced in roots and leaves especially after Pb treatment. Heterogeneous expression of CsHMA3 complemented the Pb- or Zn-sensitive phenotype of Δycf1 or Δzrc1 yeast mutant strains. Subsequently, we cloned and overexpressed CsHMA3 in Camelina. The root growth of transgenic lines was better than that in the wild-type plant under heavy metal stress (for Cd, Pb, and Zn). In particular, the transgenic lines showed enhanced Pb tolerance in a wide range of Pb concentrations. Furthermore, the Pb and Zn content in the shoots of the transgenic lines were higher than those in the wild-type plant. These results suggest that overexpression of CsHMA3 might enhance Pb and Zn tolerance and translocation. Also, the transgenic lines displayed a wider leaf shape compared with the wild-type plant due to an induction of genes related to leaf width growth and showed a greater total seed yield compared to the wild type under heavy metal stress.ConclusionsOur data obtained from physiological and functional analyses using CsHMA3 overexpression plants will be useful to develop a multifunctional plant that can improve the productivity of a bioenergy crop and simultaneously be used to purify an area contaminated by various heavy metals.

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Increased Cuticle Waxes by Overexpression of WSD1 Improves Osmotic Stress Tolerance in Arabidopsis thaliana and Camelina sativa.

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