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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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DNA-binding specificity of the GmPHD proteins.(A) Coomassie blue staining of the six GST-GmPHD fusion proteins on SDS/PAGE. Arrow indicates the fusion proteins. Lower bands probably represent the degradation products. (B) Gel shift assay of the six GmPHD proteins. GmPHD proteins (P) were incubated with a radiolabeled probe containing 5 X GTGGAG (L), in the presence (+) or absence (−) of unlabeled probes (NL) in ten-fold excess. Arrow indicates position of the protein/DNA complexes. (C) Coomassie blue staining of various domains of the GmPHD4 and GmPHD2. Arrows indicate the corresponding proteins. (D) Gel shift assay of the GmPHD4, GmPHD2 and their domains. Others are as in (B). Arrows indicate positions of the protein/DNA complexes.
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pone-0007209-g007: DNA-binding specificity of the GmPHD proteins.(A) Coomassie blue staining of the six GST-GmPHD fusion proteins on SDS/PAGE. Arrow indicates the fusion proteins. Lower bands probably represent the degradation products. (B) Gel shift assay of the six GmPHD proteins. GmPHD proteins (P) were incubated with a radiolabeled probe containing 5 X GTGGAG (L), in the presence (+) or absence (−) of unlabeled probes (NL) in ten-fold excess. Arrow indicates position of the protein/DNA complexes. (C) Coomassie blue staining of various domains of the GmPHD4 and GmPHD2. Arrows indicate the corresponding proteins. (D) Gel shift assay of the GmPHD4, GmPHD2 and their domains. Others are as in (B). Arrows indicate positions of the protein/DNA complexes.

Mentions: Alfin1, a homologue of GmPHDs from Medicago sativa, showed DNA binding activity to the conserved core of GNGGTG or GTGGNG [10]. To identify if the present GmPHDs has any DNA binding activity, we performed gel-shift analysis. Bacterially expressed GST-proteins were isolated and purified (Fig. 7A). Five tandem repeats of the sequence GTGGAG were annealed, labeled and incubated with the six purified GST-GmPHD fusion proteins. All six GmPHDs formed a complex with the labeled GTGGAG and the signal was dramatically decreased by addition of unlabeled DNA probe (Fig. 7B). These results indicate that all the six GmPHDs specifically bind to the GTGGAG element in vitro.


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)

DNA-binding specificity of the GmPHD proteins.(A) Coomassie blue staining of the six GST-GmPHD fusion proteins on SDS/PAGE. Arrow indicates the fusion proteins. Lower bands probably represent the degradation products. (B) Gel shift assay of the six GmPHD proteins. GmPHD proteins (P) were incubated with a radiolabeled probe containing 5 X GTGGAG (L), in the presence (+) or absence (−) of unlabeled probes (NL) in ten-fold excess. Arrow indicates position of the protein/DNA complexes. (C) Coomassie blue staining of various domains of the GmPHD4 and GmPHD2. Arrows indicate the corresponding proteins. (D) Gel shift assay of the GmPHD4, GmPHD2 and their domains. Others are as in (B). Arrows indicate positions of the protein/DNA complexes.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2747011&req=5

pone-0007209-g007: DNA-binding specificity of the GmPHD proteins.(A) Coomassie blue staining of the six GST-GmPHD fusion proteins on SDS/PAGE. Arrow indicates the fusion proteins. Lower bands probably represent the degradation products. (B) Gel shift assay of the six GmPHD proteins. GmPHD proteins (P) were incubated with a radiolabeled probe containing 5 X GTGGAG (L), in the presence (+) or absence (−) of unlabeled probes (NL) in ten-fold excess. Arrow indicates position of the protein/DNA complexes. (C) Coomassie blue staining of various domains of the GmPHD4 and GmPHD2. Arrows indicate the corresponding proteins. (D) Gel shift assay of the GmPHD4, GmPHD2 and their domains. Others are as in (B). Arrows indicate positions of the protein/DNA complexes.
Mentions: Alfin1, a homologue of GmPHDs from Medicago sativa, showed DNA binding activity to the conserved core of GNGGTG or GTGGNG [10]. To identify if the present GmPHDs has any DNA binding activity, we performed gel-shift analysis. Bacterially expressed GST-proteins were isolated and purified (Fig. 7A). Five tandem repeats of the sequence GTGGAG were annealed, labeled and incubated with the six purified GST-GmPHD fusion proteins. All six GmPHDs formed a complex with the labeled GTGGAG and the signal was dramatically decreased by addition of unlabeled DNA probe (Fig. 7B). These results indicate that all the six GmPHDs specifically bind to the GTGGAG element in vitro.

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