HISTONE DEACETYLASE19 is involved in jasmonic acid and ethylene signaling of pathogen response in Arabidopsis - PubMed (original) (raw)

HISTONE DEACETYLASE19 is involved in jasmonic acid and ethylene signaling of pathogen response in Arabidopsis

Changhe Zhou et al. Plant Cell. 2005 Apr.

Abstract

Histone acetylation is modulated through the action of histone acetyltransferases and deacetylases, which play key roles in the regulation of eukaryotic gene expression. Previously, we have identified a yeast histone deacetylase REDUCED POTASSIUM DEPENDENCY3 (RPD3) homolog, HISTONE DEACETYLASE19 (HDA19) (AtRPD3A), in Arabidopsis thaliana. Here, we report further study of the expression and function of HDA19. Analysis of Arabidopsis plants containing the HDA19:beta-glucuronidase fusion gene revealed that HDA19 was expressed throughout the life of the plant and in most plant organs examined. In addition, the expression of HDA19 was induced by wounding, the pathogen Alternaria brassicicola, and the plant hormones jasmonic acid and ethylene. Using green fluorescent protein fusion, we demonstrated that HDA19 accumulated in the nuclei of Arabidopsis cells. Overexpression of HDA19 in 35S:HDA19 plants decreased histone acetylation levels, whereas downregulation of HDA19 in HDA19-RNA interference (RNAi) plants increased histone acetylation levels. In comparison with wild-type plants, 35S:HDA19 transgenic plants had increased expression of ETHYLENE RESPONSE FACTOR1 and were more resistant to the pathogen A. brassicicola. The expression of jasmonic acid and ethylene regulated PATHOGENESIS-RELATED genes, Basic Chitinase and beta-1,3-Glucanase, was upregulated in 35S:HDA19 plants but downregulated in HDA19-RNAi plants. Our studies provide evidence that HDA19 may regulate gene expression involved in jasmonic acid and ethylene signaling of pathogen response in Arabidopsis.

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Figures

Figure 1.

Figure 1.

GUS Activity in HDA19:GUS Plants. GUS staining patterns of a seedling (A), leaf (B), stem (C), flower (D), stigma (E), anther (F), siliques (G), and seeds (H) from HDA19:GUS plants. GUS staining in HDA19:GUS leaves without treatment (I), treated with 1 mM methyl-jasmonate (JA) (J), 1 mM ACC (an ethylene precursor) (K), or 10 mM MgCl2 (L), infected with A. brassicicola for 24 h (M), and after cutting (N).

Figure 2.

Figure 2.

RT-PCR Analysis of Expression of HDAC Genes. Total RNA for RT-PCR analysis was isolated from leaf tissues of Arabidopsis plants treated without (control) or with 1 mM JA and 1 mM ACC (an ethylene precursor) for 6 h. Ubiquitin (UBQ) is shown as an internal control.

Figure 3.

Figure 3.

Subcellular Localization of HDA19. Protoplasts were isolated from the leaves of 35S:GFP ([A] and [B]) and 35S:HDA19-GFP ([C] and [D]) transgenic Arabidopsis plants. GFP fluorescence was examined by fluorescence microscopy under UV light ([A] and [C]) and white light ([B] and [D]).

Figure 4.

Figure 4.

Levels of Tetra-Acetylated H3 in the 35S:HDA19 and _HDA19_-RNAi Lines. (A) RNA gel blot analysis of HDA19 expression in wild-type, 35S:HDA19 (G11-1, G11-2, and G11-3), and _HDA19-_RNAi (RNAi-1 and RNAi-2) plants. Twenty micrograms of total RNA were probed with HDA19. Ethidium bromide–stained rRNA are shown to illustrate the gel loading. (B) Protein gel blot analysis detecting tetra-acetylated H3 (H3ac) (top panel) using α-H3ac antibodies on protein extracts from wild-type, 35S:HDA19 (G11-1, G11-2, and G11-3), and HDA19-RNAi (RNAi-1 and RNAi-2) plants. Bottom panel, Coomassie blue staining shows equal protein loading. MW, molecular weight marker.

Figure 5.

Figure 5.

Phenotypic Abnormalities in 35S:HDA19 Plants. (A) to (C) The 35S:HDA19 transgenic seedlings from the T2 line showed aberrant cotyledons and lacked shoot and root development ([B] and [C]) compared with a wild-type seedling (A). (D) to (F) The 35S:HDA19 transgenic plants from the T2 line with branching leaf (E) and narrow leaf (F) compared with a normal leaf (D). (G) and (H) A 6-week-old 35S:HDA19 plant (H) from the T2 line showed delayed flowering when compared with a 6-week-old wild-type plant (G). (I) and (J) A stem from a 35S:HDA19 transgenic plant of the T2 line with stunted siliques (I) and a wild-type stem with full silique elongation (J).

Figure 6.

Figure 6.

Gene Expression in the 35S:HDA19 and _HDA19_-RNAi Plants. (A) RT-PCR analysis of ERF1, BGL, and CHI-B expression in wild-type (1), 35S:HDA19 (2 and 3), and _HDA19-_RNAi (4 and 5) transgenic lines. (B) RT-PCR analysis of VSP2 and RNS1 expression in wild-type (1), 35S:HDA19 (2 and 3), and _HDA19-_RNAi (4 and 5) transgenic lines. Total RNA for RT-PCR analysis was isolated from leaf tissues of Arabidopsis plants. Ubiquitin (UBQ) is shown as an internal control.

Figure 7.

Figure 7.

Resistance to A. brassicicola by HDA19 Overexpression. Graphical representation of disease symptoms in wild-type, 35S:HDA19 (G11-1 and G11-2), and _HDA19-_RNAi (RNAi-1 and RNAi-2) transgenic lines. Disease severity index was calculated 48 and 72 h after inoculation with A. brassicicola. Disease severity index was calculated based on the degree of symptom severity as measured by leaf necrosis area: 0, no symptom; 1, 1 to 5% necrosis areas; 2, 5 to 10% necrosis areas; 3, 10 to 25% necrosis areas; 4, >25% necrosis areas. The leaf necrosis severity ratings were summed for 60 to 80 leaves of 15 to 25 plants per genotype and divided by the number of leaves rated times the maximum possible rating to given the final disease severity index. Asterisks mark values that are significantly different from the wild type (χ2 test, P < 0.05). The experiment was repeated three times with similar results.

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