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SlMYB60 regulates cuticle formation in tomato

ABSTRACT: The transcriptional regulator, SlMYB60 (Solyc10g081490.1.1) stands out as the tomato ortholog of AtMYB60, a well-characterized guard cell-specific transcription factor in Arabidopsis involved in integrating multiple stress signals in stomata. To investigate the role of SlMYB60, CRISPR-Cas9 editing was applied to two commercial tomato cultivars (Ailsa Craig and Red Setter), generating knockout alleles through multiple sgRNAs. T1 slmyb60-edited plants were assessed for stomatal aperture and drought response. Unexpectedly, mutant lines showed a slight but significative increase in stomatal pore opening in both leaves and cotyledons compared to wild type. Under dry-down conditions, leaf area, fresh weight, and relative water content were significantly affected by both genotype and treatment. RNA-seq analyses confirmed the strong downregulation of SlMYB60 in the edited lines and identified 186 differentially expressed genes (DEGs), many of which involved in cell wall or cuticle biosynthesis. Chlorophyll leaching assays indicated increased permeability in mutant lines, while electron microscopy revealed defects in epidermal cell surface structure and patterning. Notably, slmyb60 mutants exhibited thinner stomatal ledges compared to wild type, suggesting defects in stomatal architecture. Although modulating stomatal activity remains a promising strategy to improve drought resilience in tomato, our findings reveal that SlMYB60, unlike its Arabidopsis homolog, is not confined to guard cells and plays broader roles in epidermal development, indicating divergences in the MYB60 regulatory network between Arabidopsis and tomato.

ORGANISM(S): Solanum lycopersicum

PROVIDER: GSE319218 | GEO | 2026/04/22

REPOSITORIES: GEO

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Project description:Drought and salt stress severely inhibit plant growth and development. However, the regulatory mechanisms of plants in response to these stresses are not fully understood. Here we find that the expression of a WRKY transcription factor WRKY46 is rapidly induced by drought, salt and oxidative stresses. Mutations of WRKY46 by T-DNA insertion lead to more sensitive to drought and salt stress, whereas, overexpression of WRKY46 exhibits hypersensitive in soil culture with higher water loss rate, but increased tolerance on the agar plates. ABA induced stomatal closing is impaired in the WRKY46 overexpressing line (OV46), which is potentially due to the lower ROS accumulation in the guard cells. Real-time qPCR and GUS activity assay further demonstrate that WRKY46 is expressed in guard cells, but its expression is not affected by dehydration treatment, suggesting different regulatory mechanisms for WRKY46 between guard cells and other WRKY46 expressed tissues. The stomatal movement and conductance assay indicate that WRKY46 is involved in light-dependent stomatal opening. Further microarray analysis reveals that WRKY46 regulates a set of genes involved in cellular osmoprotection and redox homeostasis under dehydration stress. Determinations of ROS and MDA content confirm its role in oxidative detoxification under stress. Furthermore, we find that WRKY46 modulates light-dependent starch metabolism in guard cells via regulating QQS gene expression. Taken together, we demonstrate that WRKY46 plays a role in modulating cellular osmoprotection and redox homeostasis under drought and salt stress, and functions independently in stomatal movement via regulating light-dependent starch metabolism and ROS levels in guard cells. We used microarrays to identify the certain downstream genes regulated by WRKY46 under normal and dehydration conditions. One-week-old Arabidopsis seedlings of wild-type Col-0 (WT) and WRKY46 overexpressing line (46T) with or without dehydration treatment for 1 h were harvested and used for RNA extraction and hybridization to Affymetrix Arabidopsis ATH1 microarrays. The experiment includes 3 biological replicates.

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SlMYB60 regulates cuticle formation in tomato

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