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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.


Western blot showing the effect of PEG and HBA on histone H3 acetylation status in nuclear proteins from peanut leaves. (A) H3 acetylation status in peanut leaves treated with 20% (w/v) PEG. (B) H3 acetylation status in peanut leaves treated with 100 μM ABC. C, control group; M, Mock plants were placed in an equivalent volume of deionized water as experimental plants; 1–8 h, time point after treatment. The experiments have been carried out at least three times. Each graph displays the mean and SD of three independent experiments. */**, different from control as revealed by t-test, p < 0.05/0.01.
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Figure 1: Western blot showing the effect of PEG and HBA on histone H3 acetylation status in nuclear proteins from peanut leaves. (A) H3 acetylation status in peanut leaves treated with 20% (w/v) PEG. (B) H3 acetylation status in peanut leaves treated with 100 μM ABC. C, control group; M, Mock plants were placed in an equivalent volume of deionized water as experimental plants; 1–8 h, time point after treatment. The experiments have been carried out at least three times. Each graph displays the mean and SD of three independent experiments. */**, different from control as revealed by t-test, p < 0.05/0.01.

Mentions: The histone acetylation status of chromatin was investigated in peanut leaves subjected to PEG-induced osmotic stress or to treatment with the stress-protective hormone ABA. Immunoblotting experiments showed that the H3K9ac level increased with 20% PEG treatment, while the H3K14ac level increased with 100 μM ABA treatment (Figure 1). H3K9ac levels following PEG treatment began to increase from 2 h and continued to increase through to the 8 h time point; thus, after 5 h H3K9ac levels showed a significant increase to 7 times that of the control group, and at 8 h had increased further to 23 times control levels. Treatment with PEG produced only a marginal increase in H3K14ac levels at 8 h, but ABA treatment significantly increased the amount of H3K14ac by 5 h to 8 times that of the control group. The results indicate that both PEG and ABA can mediate changes in acetylation at different histone H3 loci; these different patterns of modification suggest that the two treatments result in different gene activation profiles in peanut leaves. At the same time, it has also been proved that the acetylation of H3K9, H3K14, and H3 were increased with 1 μM TSA treatment from 1 to 8 h (Figure S4).


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)

Western blot showing the effect of PEG and HBA on histone H3 acetylation status in nuclear proteins from peanut leaves. (A) H3 acetylation status in peanut leaves treated with 20% (w/v) PEG. (B) H3 acetylation status in peanut leaves treated with 100 μM ABC. C, control group; M, Mock plants were placed in an equivalent volume of deionized water as experimental plants; 1–8 h, time point after treatment. The experiments have been carried out at least three times. Each graph displays the mean and SD of three independent experiments. */**, different from control as revealed by t-test, p < 0.05/0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499716&req=5

Figure 1: Western blot showing the effect of PEG and HBA on histone H3 acetylation status in nuclear proteins from peanut leaves. (A) H3 acetylation status in peanut leaves treated with 20% (w/v) PEG. (B) H3 acetylation status in peanut leaves treated with 100 μM ABC. C, control group; M, Mock plants were placed in an equivalent volume of deionized water as experimental plants; 1–8 h, time point after treatment. The experiments have been carried out at least three times. Each graph displays the mean and SD of three independent experiments. */**, different from control as revealed by t-test, p < 0.05/0.01.
Mentions: The histone acetylation status of chromatin was investigated in peanut leaves subjected to PEG-induced osmotic stress or to treatment with the stress-protective hormone ABA. Immunoblotting experiments showed that the H3K9ac level increased with 20% PEG treatment, while the H3K14ac level increased with 100 μM ABA treatment (Figure 1). H3K9ac levels following PEG treatment began to increase from 2 h and continued to increase through to the 8 h time point; thus, after 5 h H3K9ac levels showed a significant increase to 7 times that of the control group, and at 8 h had increased further to 23 times control levels. Treatment with PEG produced only a marginal increase in H3K14ac levels at 8 h, but ABA treatment significantly increased the amount of H3K14ac by 5 h to 8 times that of the control group. The results indicate that both PEG and ABA can mediate changes in acetylation at different histone H3 loci; these different patterns of modification suggest that the two treatments result in different gene activation profiles in peanut leaves. At the same time, it has also been proved that the acetylation of H3K9, H3K14, and H3 were increased with 1 μM TSA treatment from 1 to 8 h (Figure S4).

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.