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Int J Mol Sci
2022 Jul 08;2314:. doi: 10.3390/ijms23147598.
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EfABI4 Transcription Factor Is Involved in the Regulation of Starch Biosynthesis in Euryale ferox Salisb Seeds.
Wu P
,
Zhu Y
,
Liu A
,
Wang Y
,
Zhao S
,
Feng K
,
Li L
.
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Starch is the final product of photosynthesis and the main storage form in plants. Studies have shown that there is a close synergistic regulatory relationship between ABA signal transduction and starch biosynthesis. In this study, we employed RNA sequencing (RNA-Seq) to investigate transcriptomic changes of the Euryale ferox seeds treated by exogenous ABA. The differentially expressed genes engaged in the "Starch and sucrose" and "TCA cycle" pathway. Furthermore, the key transcription factor EfABI4 in ABA signaling pathway and the key genes of starch biosynthesis (EfDBE1, EfSBE2, EfSS1, EfSS2, EfSS3, EfSS4 and EfGBSS1) were significantly up-regulated. Further, the Euryale ferox plant was treated with ABA, it was found that the total starch content of Euryale ferox seeds at different development stages was significantly higher than that of the control, and the key genes of starch synthesis in Euryale ferox seeds were also significantly up-regulated. Finally, yeast one-hybrid and dual luciferase assay proved that EfABI4 can promote the expression of EfSS1 by directly binding to its promoter. Subcellular localization results showed that EfABI4 protein was located at the nucleus and EfSS1 protein was located in the cytomembrane. These findings revealed that ABA promotes starch synthesis and accumulation by mediating EfABI4 to directly promote EfSS1 gene expression, which is helpful for understanding starch synthesis in seeds.
31902002 National Natural Science Foundation of China, CARS-24 China Agriculture Research System, 2020T130706 China Postdoctoral Science Foundation, JBGS [2021]017 Jiangsu seed industry revitalization'Jie Bang Gua Shuai'project
Figure 1. (A) GO enrichment of differentially expressed genes in Euryale ferox Salisb. (B) KEGG enrichment of differentially expressed genes in Euryale ferox Salisb.
Figure 2. (A): Differentially expressed gene analysis of the starch biosynthesis pathway in response to ABA. (B): The relative expression levels of EfABI4 and starch biosynthesis-related genes after ABA treatment. The ‘**’ above the histogram indicated the statistical significance at the level of 0.01 (p < 0.01).
Figure 3. (A): The effects of ABA on total starch content in E. ferox seeds. (B): Effects of ABA on amylose synthesis in E. ferox seeds. (C): Effects of ABA on amylopectin synthesis in E. ferox seeds. (D): Effect of ABA on amylose to amylopectin ratio in E. ferox seeds. (E): The relative expression levels of EfABI4, EfDBE1, EfGBSS1, EfSBE2, EfSS1, EfSS2, EfSS3
and EfSS4 after ABA treatment during Euryale ferox Salisb development. The ‘*’ or ‘**’ above the histogram indicated the statistical significance at the level of 0.05 or 0.01 (p < 0.05; p < 0.01).
Figure 4. (A): Binding activities of EfABI4 protein with the promoters of EfSS1 detected yeast one-hybrid assays. (B): The relative FLUC/RLUC activities of EfSS1 promoters activated and repressed by EfABI4 in tobacco protoplasts. The relative FLUC/RLUC activity of the negative control was set at 1. ** indicates a significant difference at p < 0.01. (C): Subcellular localization of EfSS1 and EfABI4 (EfSS1 and EfABI4 were inserted into the pCAMBIA1300-sGFP vector). Scale bars, 10 μm.
Figure 5. The model of ABA regulating starch biosynthesis in E. ferox seeds.
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