Abstract
Hypoxic environments are generally undesirable for most plants, but for astringent persimmon, high CO2 treatment (CO2 > 90%), also termed artificial high-CO2 atmosphere (AHCA), causes acetaldehyde accumulation and precipitation of soluble tannins and could remove astringency. The multiple transcriptional regulatory linkages involved in persimmon fruit deastringency have been advanced significantly by characterizing the ethylene response factors (ERFs), WRKY and MYB; however, the involvement of zinc finger proteins for deastringency has not been investigated. In this study, five genes encoding C2H2-type zinc finger proteins were isolated and designed as DkZF1-5. Phylogenetic and sequence analyses suggested the five DkZFs could be clustered into two different subgroups. qPCR analysis indicated that transcript abundances of DkZF1/4 were significantly upregulated during AHCA treatment (1% O2 and 95% CO2) at day 1, DkZF2/5 at both day 1 and 2, while DkZF3 at day 2. Dual-luciferase assay indicated DkZF1 and DkZF2 as the activators of deastringency-related structural genes (DkPDC2 and DkADH1) and transcription factors (DkERF9/10). Moreover, combinative effects between various transcription factors were investigated, indicating that DkZF1 and DkZF2 synergistically showed significantly stronger activations on the DkPDC2 promoter. Further, both bimolecular fluorescence complementation (BiFC) and yeast two hybrid (Y2H) assays confirmed that DkZF2 had protein–protein interactions with DkZF1. Thus, these findings illustrate the regulatory mechanisms of zinc finger proteins for persimmon fruit deastringency under AHCA.
Highlights
Low oxygen concentration leads to drastic metabolic rearrangements and causes rapid molecular and anaerobic responses to endure such conditions, which are mainly termed abiotic stress [1]
These oxygen levels are directly measured by the cell through sensor proteins and their target genes, and many of these sensor genes are required to maintain energy production through glycolysis, such as pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) [2,3]
The involvement of transcription factors (TFs) in artificial high-CO2 atmosphere (AHCA) driven persimmon deastringency were not fully addressed, as most of the reported TFs belonged to ethylene response factors (ERFs), NAC, MYB, WRKY, while other TFs remained unclear
Summary
Low oxygen concentration leads to drastic metabolic rearrangements and causes rapid molecular and anaerobic responses to endure such conditions, which are mainly termed abiotic stress [1]. These oxygen levels are directly measured by the cell through sensor proteins and their target genes, and many of these sensor genes are required to maintain energy production through glycolysis, such as pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) [2,3]. Protein–protein interactions of DkZFs were investigated by bimolecular fluorescence complementation (BiFC), and yeast two hybrid (Y2H) assays and their synergistic effects were analyzed
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