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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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GsGF14o overexpression promoted ABA induced stomata closure but not proline accumulation.(A) Stomata from WT and GsGF14o OX lines in response to ABA stress. Bars are 20μm. (B) Presented photos to show stomata from WT and OX lines. Bars are 10μm. (C) Percentage of different stomata in WT and OX lines. (D) Stomata aperture of WT and OX lines. *, P < 0.05; **, P < 0.01 by Student’s t test. (E) Proline content of WT and OX lines. Different letters indicated statistical differences among means by Duncan's Multiple Range Test (P < 0.05).
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pone.0146163.g009: GsGF14o overexpression promoted ABA induced stomata closure but not proline accumulation.(A) Stomata from WT and GsGF14o OX lines in response to ABA stress. Bars are 20μm. (B) Presented photos to show stomata from WT and OX lines. Bars are 10μm. (C) Percentage of different stomata in WT and OX lines. (D) Stomata aperture of WT and OX lines. *, P < 0.05; **, P < 0.01 by Student’s t test. (E) Proline content of WT and OX lines. Different letters indicated statistical differences among means by Duncan's Multiple Range Test (P < 0.05).

Mentions: In addition to the inhibition on seed germination and seedling growth, another two typical effects of ABA on plants are the induced stomata closure and proline accumulation [13]. Our previous study showed that overexpression of GsGF14o in Arabidopsis resulted in smaller stomata, as evidenced by a decrease of the stomata length and width [37]. In order to test whether GsGF14o overexpression affected ABA induced stomata closure, we measured the numbers of completely open, partially open and completely closed stomata (Fig 9), respectively, by using scanning electron microscopy. As shown in Fig 9A and 9B, under ABA treatment, the transgenic lines displayed much less open stomata, but more closed stomata than WT. Statistically speaking, the percentage of completely open stomata was 29% for WT, but these values decreased to 15.2% for line 1, 12.5% for line 4, and 7.9% for line 9. Notably, line 9 displayed higher percentage of partially open stomata than WT, while line 1 and 4 showed more completely closed stomata (Fig 9C). These data illustrated that GsGF14o overexpression in Arabidopsis promoted ABA induced stomata closure. To confirm this, we further determined stomata apertures (presented by the rates of width over length) of WT and OX lines under ABA treatment. As expected, statistical analysis also suggested that compared with WT, stomata aperture of transgenic lines was more sensitive to ABA (Fig 9D).


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)

GsGF14o overexpression promoted ABA induced stomata closure but not proline accumulation.(A) Stomata from WT and GsGF14o OX lines in response to ABA stress. Bars are 20μm. (B) Presented photos to show stomata from WT and OX lines. Bars are 10μm. (C) Percentage of different stomata in WT and OX lines. (D) Stomata aperture of WT and OX lines. *, P < 0.05; **, P < 0.01 by Student’s t test. (E) Proline content of WT and OX lines. Different letters indicated statistical differences among means by Duncan's Multiple Range Test (P < 0.05).
© Copyright Policy
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pone.0146163.g009: GsGF14o overexpression promoted ABA induced stomata closure but not proline accumulation.(A) Stomata from WT and GsGF14o OX lines in response to ABA stress. Bars are 20μm. (B) Presented photos to show stomata from WT and OX lines. Bars are 10μm. (C) Percentage of different stomata in WT and OX lines. (D) Stomata aperture of WT and OX lines. *, P < 0.05; **, P < 0.01 by Student’s t test. (E) Proline content of WT and OX lines. Different letters indicated statistical differences among means by Duncan's Multiple Range Test (P < 0.05).
Mentions: In addition to the inhibition on seed germination and seedling growth, another two typical effects of ABA on plants are the induced stomata closure and proline accumulation [13]. Our previous study showed that overexpression of GsGF14o in Arabidopsis resulted in smaller stomata, as evidenced by a decrease of the stomata length and width [37]. In order to test whether GsGF14o overexpression affected ABA induced stomata closure, we measured the numbers of completely open, partially open and completely closed stomata (Fig 9), respectively, by using scanning electron microscopy. As shown in Fig 9A and 9B, under ABA treatment, the transgenic lines displayed much less open stomata, but more closed stomata than WT. Statistically speaking, the percentage of completely open stomata was 29% for WT, but these values decreased to 15.2% for line 1, 12.5% for line 4, and 7.9% for line 9. Notably, line 9 displayed higher percentage of partially open stomata than WT, while line 1 and 4 showed more completely closed stomata (Fig 9C). These data illustrated that GsGF14o overexpression in Arabidopsis promoted ABA induced stomata closure. To confirm this, we further determined stomata apertures (presented by the rates of width over length) of WT and OX lines under ABA treatment. As expected, statistical analysis also suggested that compared with WT, stomata aperture of transgenic lines was more sensitive to ABA (Fig 9D).

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