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A 14-3-3 Family Protein from Wild Soybean (Glycine Soja) Regulates ABA Sensitivity in Arabidopsis.

Sun X, Sun M, Jia B, Chen C, Qin Z, Yang K, Shen Y, Meiping Z, Mingyang C, Zhu Y - PLoS ONE (2015)

Bottom Line: Then, based on our previous functional characterization of a Glycine soja 14-3-3 protein GsGF14o in drought stress responses, we further investigated the expression characteristics of GsGF14o in detail, and demonstrated its positive roles in ABA sensitivity.As expected, GsGF14o overexpression in Arabidopsis augmented the ABA inhibition of seed germination and seedling growth, promoted the ABA induced stomata closure, and up-regulated the expression levels of ABA induced genes.Taken together, results presented in this study strongly suggested that GsGF14o played an important role in regulation of ABA sensitivity in Arabidopsis.

View Article: PubMed Central - PubMed

Affiliation: Crop Stress Molecular Biology Laboratory, Heilongjiang Bayi Agricultural University, Daqing, P.R. China.

ABSTRACT
It is widely accepted that the 14-3-3 family proteins are key regulators of multiple stress signal transduction cascades. By conducting genome-wide analysis, researchers have identified the soybean 14-3-3 family proteins; however, until now, there is still no direct genetic evidence showing the involvement of soybean 14-3-3s in ABA responses. Hence, in this study, based on the latest Glycine max genome on Phytozome v10.3, we initially analyzed the evolutionary relationship, genome organization, gene structure and duplication, and three-dimensional structure of soybean 14-3-3 family proteins systematically. Our results suggested that soybean 14-3-3 family was highly evolutionary conserved and possessed segmental duplication in evolution. Then, based on our previous functional characterization of a Glycine soja 14-3-3 protein GsGF14o in drought stress responses, we further investigated the expression characteristics of GsGF14o in detail, and demonstrated its positive roles in ABA sensitivity. Quantitative real-time PCR analyses in Glycine soja seedlings and GUS activity assays in PGsGF14O:GUS transgenic Arabidopsis showed that GsGF14o expression was moderately and rapidly induced by ABA treatment. As expected, GsGF14o overexpression in Arabidopsis augmented the ABA inhibition of seed germination and seedling growth, promoted the ABA induced stomata closure, and up-regulated the expression levels of ABA induced genes. Moreover, through yeast two hybrid analyses, we further demonstrated that GsGF14o physically interacted with the AREB/ABF transcription factors in yeast cells. Taken together, results presented in this study strongly suggested that GsGF14o played an important role in regulation of ABA sensitivity in Arabidopsis.

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Protein interaction of GsGF14o with ABF transcription factors in yeast cells.(A) Schematic representation of GsGF14o-BD and ABFs-AD fused expression constructors. (B) Yeast two hybrid identification of protein interaction between GsGF14o and ABF transcription factors. The GsGF14o-BD/AD combination was used as a negative control.
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pone.0146163.g011: Protein interaction of GsGF14o with ABF transcription factors in yeast cells.(A) Schematic representation of GsGF14o-BD and ABFs-AD fused expression constructors. (B) Yeast two hybrid identification of protein interaction between GsGF14o and ABF transcription factors. The GsGF14o-BD/AD combination was used as a negative control.

Mentions: Recent studies uncovered that 14-3-3 proteins could physically interact with the AREB/ABF transcription factors, and participate in the ABA signaling transduction in plant cells [29,31,36]. Considering the great changes in expression of these ABRE-containing genes described above (Fig 10), we speculated that GsGF14o might also interact with the AREB/ABF transcription factors. To verify this hypothesis, we used the Y2H (Yeast Two Hybrid) technology to determine whether GsGF14o could interact with AREB/ABFs as described previously [32]. To do this, the Arabidopsis ABF1, ABF2, ABF3, ABF4 and ABI5 were in-fused cloned to the pGADT7 vector to express AREB/ABFs at the C-terminus of GAL4 activating domain (Fig 11A). The full-length GsGF14o gene was fused to the GAL4 DNA-binding domain in the pGBKT7 vector, and used as a bait to analyze the protein interaction with AREB/ABFs (Fig 11A). Y2H assays showed that the recombinant yeast cells harboring the pGBKT7-GsGF14o and pGADT7-ABFs vectors could survive well on the SD/-T-L, SD/-T-L-H, and SD/-T-L-H-A selective medium (Fig 11B). However, the yeast cells carrying the pGBKT7-GsGF14o and empty pGADT7 vectors only showed growth on the SD/-T-L medium, but not on the SD/-T-L-H and SD/-T-L-H-A medium. Taken together, these results suggested that GsGF14o did physically interact with AREB/ABFs in yeast cells.


A 14-3-3 Family Protein from Wild Soybean (Glycine Soja) Regulates ABA Sensitivity in Arabidopsis.

Sun X, Sun M, Jia B, Chen C, Qin Z, Yang K, Shen Y, Meiping Z, Mingyang C, Zhu Y - PLoS ONE (2015)

Protein interaction of GsGF14o with ABF transcription factors in yeast cells.(A) Schematic representation of GsGF14o-BD and ABFs-AD fused expression constructors. (B) Yeast two hybrid identification of protein interaction between GsGF14o and ABF transcription factors. The GsGF14o-BD/AD combination was used as a negative control.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4696740&req=5

pone.0146163.g011: Protein interaction of GsGF14o with ABF transcription factors in yeast cells.(A) Schematic representation of GsGF14o-BD and ABFs-AD fused expression constructors. (B) Yeast two hybrid identification of protein interaction between GsGF14o and ABF transcription factors. The GsGF14o-BD/AD combination was used as a negative control.
Mentions: Recent studies uncovered that 14-3-3 proteins could physically interact with the AREB/ABF transcription factors, and participate in the ABA signaling transduction in plant cells [29,31,36]. Considering the great changes in expression of these ABRE-containing genes described above (Fig 10), we speculated that GsGF14o might also interact with the AREB/ABF transcription factors. To verify this hypothesis, we used the Y2H (Yeast Two Hybrid) technology to determine whether GsGF14o could interact with AREB/ABFs as described previously [32]. To do this, the Arabidopsis ABF1, ABF2, ABF3, ABF4 and ABI5 were in-fused cloned to the pGADT7 vector to express AREB/ABFs at the C-terminus of GAL4 activating domain (Fig 11A). The full-length GsGF14o gene was fused to the GAL4 DNA-binding domain in the pGBKT7 vector, and used as a bait to analyze the protein interaction with AREB/ABFs (Fig 11A). Y2H assays showed that the recombinant yeast cells harboring the pGBKT7-GsGF14o and pGADT7-ABFs vectors could survive well on the SD/-T-L, SD/-T-L-H, and SD/-T-L-H-A selective medium (Fig 11B). However, the yeast cells carrying the pGBKT7-GsGF14o and empty pGADT7 vectors only showed growth on the SD/-T-L medium, but not on the SD/-T-L-H and SD/-T-L-H-A medium. Taken together, these results suggested that GsGF14o did physically interact with AREB/ABFs in yeast cells.

Bottom Line: Then, based on our previous functional characterization of a Glycine soja 14-3-3 protein GsGF14o in drought stress responses, we further investigated the expression characteristics of GsGF14o in detail, and demonstrated its positive roles in ABA sensitivity.As expected, GsGF14o overexpression in Arabidopsis augmented the ABA inhibition of seed germination and seedling growth, promoted the ABA induced stomata closure, and up-regulated the expression levels of ABA induced genes.Taken together, results presented in this study strongly suggested that GsGF14o played an important role in regulation of ABA sensitivity in Arabidopsis.

View Article: PubMed Central - PubMed

Affiliation: Crop Stress Molecular Biology Laboratory, Heilongjiang Bayi Agricultural University, Daqing, P.R. China.

ABSTRACT
It is widely accepted that the 14-3-3 family proteins are key regulators of multiple stress signal transduction cascades. By conducting genome-wide analysis, researchers have identified the soybean 14-3-3 family proteins; however, until now, there is still no direct genetic evidence showing the involvement of soybean 14-3-3s in ABA responses. Hence, in this study, based on the latest Glycine max genome on Phytozome v10.3, we initially analyzed the evolutionary relationship, genome organization, gene structure and duplication, and three-dimensional structure of soybean 14-3-3 family proteins systematically. Our results suggested that soybean 14-3-3 family was highly evolutionary conserved and possessed segmental duplication in evolution. Then, based on our previous functional characterization of a Glycine soja 14-3-3 protein GsGF14o in drought stress responses, we further investigated the expression characteristics of GsGF14o in detail, and demonstrated its positive roles in ABA sensitivity. Quantitative real-time PCR analyses in Glycine soja seedlings and GUS activity assays in PGsGF14O:GUS transgenic Arabidopsis showed that GsGF14o expression was moderately and rapidly induced by ABA treatment. As expected, GsGF14o overexpression in Arabidopsis augmented the ABA inhibition of seed germination and seedling growth, promoted the ABA induced stomata closure, and up-regulated the expression levels of ABA induced genes. Moreover, through yeast two hybrid analyses, we further demonstrated that GsGF14o physically interacted with the AREB/ABF transcription factors in yeast cells. Taken together, results presented in this study strongly suggested that GsGF14o played an important role in regulation of ABA sensitivity in Arabidopsis.

Show MeSH
Related in: MedlinePlus