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Soybean GmPHD-type transcription regulators improve stress tolerance in transgenic Arabidopsis plants.

Wei W, Huang J, Hao YJ, Zou HF, Wang HW, Zhao JY, Liu XY, Zhang WK, Ma B, Zhang JS, Chen SY - PLoS ONE (2009)

Bottom Line: Transgenic Arabidopsis plants overexpressing the GmPHD2 showed salt tolerance when compared with the wild type plants.This tolerance was likely achieved by diminishing the oxidative stress through regulation of downstream genes.These results provide important clues for soybean stress tolerance through manipulation of PHD-type transcription regulator.

View Article: PubMed Central - PubMed

Affiliation: Plant Gene Research Center, National Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT

Background: Soybean [Glycine max (L.) Merr.] is one of the most important crops for oil and protein resource. Improvement of stress tolerance will be beneficial for soybean seed production.

Principal findings: Six GmPHD genes encoding Alfin1-type PHD finger protein were identified and their expressions differentially responded to drought, salt, cold and ABA treatments. The six GmPHDs were nuclear proteins and showed ability to bind the cis-element "GTGGAG". The N-terminal domain of GmPHD played a major role in DNA binding. Using a protoplast assay system, we find that GmPHD1 to GmPHD5 had transcriptional suppression activity whereas GmPHD6 did not have. In yeast assay, the GmPHD6 can form homodimer and heterodimer with the other GmPHDs except GmPHD2. The N-terminal plus the variable regions but not the PHD-finger is required for the dimerization. Transgenic Arabidopsis plants overexpressing the GmPHD2 showed salt tolerance when compared with the wild type plants. This tolerance was likely achieved by diminishing the oxidative stress through regulation of downstream genes.

Significance: These results provide important clues for soybean stress tolerance through manipulation of PHD-type transcription regulator.

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Transcriptional regulation activity of GmPHDs in protoplast assay.(A) Effects of the GmPHDs on reporter gene expression as revealed by relative LUC activity. The GAL4 DNA-binding domain (BD) and VP16 were used as negative and positive controls respectively. “**” indicate highly significant difference (P<0.01) compared to BD value. (B) Effects of the GmPHDs on VP16-mediated LUC gene expression. The Arabiodpsis Dof23 was used as a non-interactive control. (C) Effects of various domains of the GmPHD2 on VP16-mediated LUC gene expression. For (B) and (C), “*” and “**” indicate significant difference (P<0.05 and P<0.01 respectively) compared to VP16 value.
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pone-0007209-g005: Transcriptional regulation activity of GmPHDs in protoplast assay.(A) Effects of the GmPHDs on reporter gene expression as revealed by relative LUC activity. The GAL4 DNA-binding domain (BD) and VP16 were used as negative and positive controls respectively. “**” indicate highly significant difference (P<0.01) compared to BD value. (B) Effects of the GmPHDs on VP16-mediated LUC gene expression. The Arabiodpsis Dof23 was used as a non-interactive control. (C) Effects of various domains of the GmPHD2 on VP16-mediated LUC gene expression. For (B) and (C), “*” and “**” indicate significant difference (P<0.05 and P<0.01 respectively) compared to VP16 value.

Mentions: The PHD finger proteins have been reported to have the transcriptional activation activity [36]. We examined the transcriptional activation activity of GmPHDs in protoplast system. As shown in Fig. 5A, among the six proteins compared, five (except the GmPHD6) was found to have inhibitory effect on reporter gene activity when compared to the negative BD control, possibly implying that the five proteins GmPHD1 to GmPHD5 can suppress the transcription of the reporter gene to different degrees. The GmPHD6 appeared not to have such inhibitory activity. To further investigate if the GmPHD proteins have any effect on VP16-mediated transcriptional activation, we included each of the six GmPHD proteins with the positive control VP16 transcription factor in the assay system. Fig. 5B showed that the five proteins GmPHD1 to GmPHD5 had inhibitory effects on VP16-promoted gene expression, suggesting that the five proteins may mainly play roles in transcriptional suppression. On the contrary, the GmPHD6 did not show such ability. A Dof-type transcription factor Dof23 from Arabidopsis did not have significant effect on VP16 transactivation activity.


Soybean GmPHD-type transcription regulators improve stress tolerance in transgenic Arabidopsis plants.

Wei W, Huang J, Hao YJ, Zou HF, Wang HW, Zhao JY, Liu XY, Zhang WK, Ma B, Zhang JS, Chen SY - PLoS ONE (2009)

Transcriptional regulation activity of GmPHDs in protoplast assay.(A) Effects of the GmPHDs on reporter gene expression as revealed by relative LUC activity. The GAL4 DNA-binding domain (BD) and VP16 were used as negative and positive controls respectively. “**” indicate highly significant difference (P<0.01) compared to BD value. (B) Effects of the GmPHDs on VP16-mediated LUC gene expression. The Arabiodpsis Dof23 was used as a non-interactive control. (C) Effects of various domains of the GmPHD2 on VP16-mediated LUC gene expression. For (B) and (C), “*” and “**” indicate significant difference (P<0.05 and P<0.01 respectively) compared to VP16 value.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2747011&req=5

pone-0007209-g005: Transcriptional regulation activity of GmPHDs in protoplast assay.(A) Effects of the GmPHDs on reporter gene expression as revealed by relative LUC activity. The GAL4 DNA-binding domain (BD) and VP16 were used as negative and positive controls respectively. “**” indicate highly significant difference (P<0.01) compared to BD value. (B) Effects of the GmPHDs on VP16-mediated LUC gene expression. The Arabiodpsis Dof23 was used as a non-interactive control. (C) Effects of various domains of the GmPHD2 on VP16-mediated LUC gene expression. For (B) and (C), “*” and “**” indicate significant difference (P<0.05 and P<0.01 respectively) compared to VP16 value.
Mentions: The PHD finger proteins have been reported to have the transcriptional activation activity [36]. We examined the transcriptional activation activity of GmPHDs in protoplast system. As shown in Fig. 5A, among the six proteins compared, five (except the GmPHD6) was found to have inhibitory effect on reporter gene activity when compared to the negative BD control, possibly implying that the five proteins GmPHD1 to GmPHD5 can suppress the transcription of the reporter gene to different degrees. The GmPHD6 appeared not to have such inhibitory activity. To further investigate if the GmPHD proteins have any effect on VP16-mediated transcriptional activation, we included each of the six GmPHD proteins with the positive control VP16 transcription factor in the assay system. Fig. 5B showed that the five proteins GmPHD1 to GmPHD5 had inhibitory effects on VP16-promoted gene expression, suggesting that the five proteins may mainly play roles in transcriptional suppression. On the contrary, the GmPHD6 did not show such ability. A Dof-type transcription factor Dof23 from Arabidopsis did not have significant effect on VP16 transactivation activity.

Bottom Line: Transgenic Arabidopsis plants overexpressing the GmPHD2 showed salt tolerance when compared with the wild type plants.This tolerance was likely achieved by diminishing the oxidative stress through regulation of downstream genes.These results provide important clues for soybean stress tolerance through manipulation of PHD-type transcription regulator.

View Article: PubMed Central - PubMed

Affiliation: Plant Gene Research Center, National Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT

Background: Soybean [Glycine max (L.) Merr.] is one of the most important crops for oil and protein resource. Improvement of stress tolerance will be beneficial for soybean seed production.

Principal findings: Six GmPHD genes encoding Alfin1-type PHD finger protein were identified and their expressions differentially responded to drought, salt, cold and ABA treatments. The six GmPHDs were nuclear proteins and showed ability to bind the cis-element "GTGGAG". The N-terminal domain of GmPHD played a major role in DNA binding. Using a protoplast assay system, we find that GmPHD1 to GmPHD5 had transcriptional suppression activity whereas GmPHD6 did not have. In yeast assay, the GmPHD6 can form homodimer and heterodimer with the other GmPHDs except GmPHD2. The N-terminal plus the variable regions but not the PHD-finger is required for the dimerization. Transgenic Arabidopsis plants overexpressing the GmPHD2 showed salt tolerance when compared with the wild type plants. This tolerance was likely achieved by diminishing the oxidative stress through regulation of downstream genes.

Significance: These results provide important clues for soybean stress tolerance through manipulation of PHD-type transcription regulator.

Show MeSH
Related in: MedlinePlus