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Isolation and characterization of an osmotic stress and ABA induced histone deacetylase in Arachis hygogaea.

Su LC, Deng B, Liu S, Li LM, Hu B, Zhong YT, Li L - Front Plant Sci (2015)

Bottom Line: Using RNA-seq data for peanut, we found a RPD3/HDA1-like superfamily histone deacetylase (HDAC), termed AhHDA1, whose gene is up-regulated by PEG-induced water limitation and ABA signaling.To understand whether and how osmotic stress and ABA mediate the peanut stress response by epigenetics, the expression of AhHDA1 and stress-responsive genes following treatment with PEG, ABA, and the specific HDAC inhibitor trichostatin A (TSA) were analyzed.AhHDA1 transcript levels were enhanced by all three treatments, as was expression of peanut transcription factor genes, indicating that AhHDA1 might be involved in the epigenetic regulation of stress resistance genes that comprise the responses to osmotic stress and ABA.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University Guangzhou, China.

ABSTRACT
Histone acetylation, which together with histone methylation regulates gene activity in response to stress, is an important epigenetic modification. There is an increasing research focus on histone acetylation in crops, but there is no information to date in peanut (Arachis hypogaea). We showed that osmotic stress and ABA affect the acetylation of histone H3 loci in peanut seedlings by immunoblotting experiments. Using RNA-seq data for peanut, we found a RPD3/HDA1-like superfamily histone deacetylase (HDAC), termed AhHDA1, whose gene is up-regulated by PEG-induced water limitation and ABA signaling. We isolated and characterized AhHDA1 from A. hypogaea, showing that AhHDA1 is very similar to an Arabidopsis HDAC (AtHDA6) and, in recombinant form, possesses HDAC activity. To understand whether and how osmotic stress and ABA mediate the peanut stress response by epigenetics, the expression of AhHDA1 and stress-responsive genes following treatment with PEG, ABA, and the specific HDAC inhibitor trichostatin A (TSA) were analyzed. AhHDA1 transcript levels were enhanced by all three treatments, as was expression of peanut transcription factor genes, indicating that AhHDA1 might be involved in the epigenetic regulation of stress resistance genes that comprise the responses to osmotic stress and ABA.

No MeSH data available.


Relatedness of peanut HDACs sequences to counterparts in other plants. (A) Alignment of deduced amino acid sequence of peanut HDACs with other plant HDAC sequences. The degree of similarity of the amino acid residues at each residues at each aligned position is shaded black, red, blue, in decreasing order. GenBank accession numbers for each aligned HDAC protein are indicated in parenthesis. (B) Phylogenetic analysis of amino acid sequences of AhHDA1 and other plant HDACs. A multiple sequence alignment was performed using Clustal W and the phylogenetic tree was constructed via the Neighbor-Joining method in MEGA 4 software. Bootstrap values from 1000 replicates for each branch are shown. GenBank accession numbers: Glycine max HDA6 (XP_003525556.1), Phaseolus vulgaris HAD (XP_007155467.1), Arachis hypogaea HDA1 (JR541338.1), Medicago truncatula HDA (XP_003601202.1), Prunus persica HDA (XP_007209104.1), Populus euphartica HDA (XP_011046214.1). Cucumis melo HDA6 (XP_00864523.1), Cucumis sativas HDA6 (XP_004138094.1), Citrus sinensis HDA6 (XP_006476865.1), Arabidopsis thaliana HA6 (AED97705.1), Gossypium arboretum HDA (KHG12201.1), Theobroma cacao HDA (XP_007036337.1), Arachis hypogaea HDA19-like (AHA85936.1), Arachis hypogaea HDA15-like (AHA85936.1). The scale bar is 0.02.
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Figure 2: Relatedness of peanut HDACs sequences to counterparts in other plants. (A) Alignment of deduced amino acid sequence of peanut HDACs with other plant HDAC sequences. The degree of similarity of the amino acid residues at each residues at each aligned position is shaded black, red, blue, in decreasing order. GenBank accession numbers for each aligned HDAC protein are indicated in parenthesis. (B) Phylogenetic analysis of amino acid sequences of AhHDA1 and other plant HDACs. A multiple sequence alignment was performed using Clustal W and the phylogenetic tree was constructed via the Neighbor-Joining method in MEGA 4 software. Bootstrap values from 1000 replicates for each branch are shown. GenBank accession numbers: Glycine max HDA6 (XP_003525556.1), Phaseolus vulgaris HAD (XP_007155467.1), Arachis hypogaea HDA1 (JR541338.1), Medicago truncatula HDA (XP_003601202.1), Prunus persica HDA (XP_007209104.1), Populus euphartica HDA (XP_011046214.1). Cucumis melo HDA6 (XP_00864523.1), Cucumis sativas HDA6 (XP_004138094.1), Citrus sinensis HDA6 (XP_006476865.1), Arabidopsis thaliana HA6 (AED97705.1), Gossypium arboretum HDA (KHG12201.1), Theobroma cacao HDA (XP_007036337.1), Arachis hypogaea HDA19-like (AHA85936.1), Arachis hypogaea HDA15-like (AHA85936.1). The scale bar is 0.02.

Mentions: Specific forward and reverse primers (ORF1-F and ORF1-R) were designed from comp66763_c0 to isolate an AhHDA1 cDNA as detailed in Materials and Methods. By sequence alignment, the predicted sequence of the AhHDA1 protein showed a high degree of similarity with other HDACs in the GenBank DNA database, and AhHDA1 possessed the same active site and Zn2+ binding sites as other plant HDACs (Figure 2A). AhHDA1 consists of a polypeptide of 467 amino acid residues with a calculated molecular weight of 52.37 kDa and an isoelectric point of 5.28. It can be deduced from Figure 2B that AhHDA1 is most similar to counterparts in eudicots, especially soybean.


Isolation and characterization of an osmotic stress and ABA induced histone deacetylase in Arachis hygogaea.

Su LC, Deng B, Liu S, Li LM, Hu B, Zhong YT, Li L - Front Plant Sci (2015)

Relatedness of peanut HDACs sequences to counterparts in other plants. (A) Alignment of deduced amino acid sequence of peanut HDACs with other plant HDAC sequences. The degree of similarity of the amino acid residues at each residues at each aligned position is shaded black, red, blue, in decreasing order. GenBank accession numbers for each aligned HDAC protein are indicated in parenthesis. (B) Phylogenetic analysis of amino acid sequences of AhHDA1 and other plant HDACs. A multiple sequence alignment was performed using Clustal W and the phylogenetic tree was constructed via the Neighbor-Joining method in MEGA 4 software. Bootstrap values from 1000 replicates for each branch are shown. GenBank accession numbers: Glycine max HDA6 (XP_003525556.1), Phaseolus vulgaris HAD (XP_007155467.1), Arachis hypogaea HDA1 (JR541338.1), Medicago truncatula HDA (XP_003601202.1), Prunus persica HDA (XP_007209104.1), Populus euphartica HDA (XP_011046214.1). Cucumis melo HDA6 (XP_00864523.1), Cucumis sativas HDA6 (XP_004138094.1), Citrus sinensis HDA6 (XP_006476865.1), Arabidopsis thaliana HA6 (AED97705.1), Gossypium arboretum HDA (KHG12201.1), Theobroma cacao HDA (XP_007036337.1), Arachis hypogaea HDA19-like (AHA85936.1), Arachis hypogaea HDA15-like (AHA85936.1). The scale bar is 0.02.
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Figure 2: Relatedness of peanut HDACs sequences to counterparts in other plants. (A) Alignment of deduced amino acid sequence of peanut HDACs with other plant HDAC sequences. The degree of similarity of the amino acid residues at each residues at each aligned position is shaded black, red, blue, in decreasing order. GenBank accession numbers for each aligned HDAC protein are indicated in parenthesis. (B) Phylogenetic analysis of amino acid sequences of AhHDA1 and other plant HDACs. A multiple sequence alignment was performed using Clustal W and the phylogenetic tree was constructed via the Neighbor-Joining method in MEGA 4 software. Bootstrap values from 1000 replicates for each branch are shown. GenBank accession numbers: Glycine max HDA6 (XP_003525556.1), Phaseolus vulgaris HAD (XP_007155467.1), Arachis hypogaea HDA1 (JR541338.1), Medicago truncatula HDA (XP_003601202.1), Prunus persica HDA (XP_007209104.1), Populus euphartica HDA (XP_011046214.1). Cucumis melo HDA6 (XP_00864523.1), Cucumis sativas HDA6 (XP_004138094.1), Citrus sinensis HDA6 (XP_006476865.1), Arabidopsis thaliana HA6 (AED97705.1), Gossypium arboretum HDA (KHG12201.1), Theobroma cacao HDA (XP_007036337.1), Arachis hypogaea HDA19-like (AHA85936.1), Arachis hypogaea HDA15-like (AHA85936.1). The scale bar is 0.02.
Mentions: Specific forward and reverse primers (ORF1-F and ORF1-R) were designed from comp66763_c0 to isolate an AhHDA1 cDNA as detailed in Materials and Methods. By sequence alignment, the predicted sequence of the AhHDA1 protein showed a high degree of similarity with other HDACs in the GenBank DNA database, and AhHDA1 possessed the same active site and Zn2+ binding sites as other plant HDACs (Figure 2A). AhHDA1 consists of a polypeptide of 467 amino acid residues with a calculated molecular weight of 52.37 kDa and an isoelectric point of 5.28. It can be deduced from Figure 2B that AhHDA1 is most similar to counterparts in eudicots, especially soybean.

Bottom Line: Using RNA-seq data for peanut, we found a RPD3/HDA1-like superfamily histone deacetylase (HDAC), termed AhHDA1, whose gene is up-regulated by PEG-induced water limitation and ABA signaling.To understand whether and how osmotic stress and ABA mediate the peanut stress response by epigenetics, the expression of AhHDA1 and stress-responsive genes following treatment with PEG, ABA, and the specific HDAC inhibitor trichostatin A (TSA) were analyzed.AhHDA1 transcript levels were enhanced by all three treatments, as was expression of peanut transcription factor genes, indicating that AhHDA1 might be involved in the epigenetic regulation of stress resistance genes that comprise the responses to osmotic stress and ABA.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University Guangzhou, China.

ABSTRACT
Histone acetylation, which together with histone methylation regulates gene activity in response to stress, is an important epigenetic modification. There is an increasing research focus on histone acetylation in crops, but there is no information to date in peanut (Arachis hypogaea). We showed that osmotic stress and ABA affect the acetylation of histone H3 loci in peanut seedlings by immunoblotting experiments. Using RNA-seq data for peanut, we found a RPD3/HDA1-like superfamily histone deacetylase (HDAC), termed AhHDA1, whose gene is up-regulated by PEG-induced water limitation and ABA signaling. We isolated and characterized AhHDA1 from A. hypogaea, showing that AhHDA1 is very similar to an Arabidopsis HDAC (AtHDA6) and, in recombinant form, possesses HDAC activity. To understand whether and how osmotic stress and ABA mediate the peanut stress response by epigenetics, the expression of AhHDA1 and stress-responsive genes following treatment with PEG, ABA, and the specific HDAC inhibitor trichostatin A (TSA) were analyzed. AhHDA1 transcript levels were enhanced by all three treatments, as was expression of peanut transcription factor genes, indicating that AhHDA1 might be involved in the epigenetic regulation of stress resistance genes that comprise the responses to osmotic stress and ABA.

No MeSH data available.