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The Hv-SGT1 gene from Haynaldia villosa contributes to resistances towards both biotrophic and hemi-biotrophic pathogens in common wheat (Triticum aestivum L.).

Xing L, Qian C, Cao A, Li Y, Jiang Z, Li M, Jin X, Hu J, Zhang Y, Wang X, Chen P - PLoS ONE (2013)

Bottom Line: The demonstration that silencing of Hv-SGT1 substantially reduced resistance to Bgt indicated that Hv-SGT1 was an essential component of disease resistance in H. villosa.Therefore, the involvement of Hv-SGT1 in H2O2 production correlates with the hypersensitive response and jasmonic acid signaling.Our novel demonstration that wheat with over-expressed Hv-SGT1 showed enhanced resistance to both powdery mildew and FHB suggests that it could served as a transgenic genetic resource in wheat breeding for multiple disease resistance.

View Article: PubMed Central - PubMed

Affiliation: The National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
The SGT1 protein is essential for R protein-mediated and PAMPs-triggered resistance in many plant species. Here we reported the isolation and characterization of the Hv-SGT1 gene from Haynaldiavillosa (2n = 14, VV). Analysis of the subcellular location of Hv-SGT1 by transient expression of a fusion to GFP indicated its presence in the cytoplasm and nucleus. Levels of Hv-SGT1 transcripts were increased by inoculation with either the biotrophic pathogen Blumeriagraminis DC. f. Sp. tritici (Bgt) or the hemi-biotrophic pathogen Fusariumgraminearum (Fg). Levels of Hv-SGT1 showed substantial increase following treatment with H2O2 and methyl jasmonate (MeJA), only slightly induced following exposure to ethephon or abscisic acid, but not changed following exposure to salicylic acid. The demonstration that silencing of Hv-SGT1 substantially reduced resistance to Bgt indicated that Hv-SGT1 was an essential component of disease resistance in H. villosa. The over-expression of Hv-SGT1 in Yangmai 158 enhanced resistance to powdery mildew, and this correlated with increased levels of whole-cell reactive oxygen intermediates at the sites of penetration by the pathogens. Compared with wild-type plants, the expression levels of genes related to the H2O2 and JA signaling pathways were lower in the Hv-SGT1 silenced plants and higher in the Hv-SGT1 over-expressing plants. Therefore, the involvement of Hv-SGT1 in H2O2 production correlates with the hypersensitive response and jasmonic acid signaling. Our novel demonstration that wheat with over-expressed Hv-SGT1 showed enhanced resistance to both powdery mildew and FHB suggests that it could served as a transgenic genetic resource in wheat breeding for multiple disease resistance.

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Functional analysis of the Hv-SGT1 gene by BSMV-induced gene silencing.(A) Following infection of H. villosa with the virus, chlorotic mosaic symptoms were observed on the 4th leaves 10 days after inoculation (dpi) with BSMV:γ or BSMV:Hv-SGT1. Photobleaching was observed on leaves infected with BSMV:PDS 15 dpi, whereas no detectable phenotype was observed in the mock-treated plants inoculated with 1×GKP buffer. Representative photographs were taken 15 days after virus inoculation. (B) The Hv-SGT1 gene was efficiently silenced in the BSMV: Hv-SGT1 inoculated leaves compared to the control inoculated with BSMV:γ, as indicated using qRT-PCR analysis. (C) More infected Bgt could developed into secondary hyphae in the BSMV: Hv-SGT1 inoculated leaves compared to the BSMV:γ infected leaves 10 days after inoculation with Bgt. (D) The rate of formation of secondary hyphae (SH) was higher in the Hv-SGT1 silenced leaves inoculated with BSMV:Hv-SGT1 compared with that in BSMV:γ-infected leaves at 5, 7, and 10 days after inoculation with Bgt. (E) Levels of NADPHOX, APX, NPR1, PR5, OPDA, COI1, ERF1, PR10, PR2, PR3, GSTand GPOXin Hv-SGT1 silenced leaves. * p < 0.05, ** p < 0.01 compared to the BSMV:γ-infected leaves.
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pone-0072571-g005: Functional analysis of the Hv-SGT1 gene by BSMV-induced gene silencing.(A) Following infection of H. villosa with the virus, chlorotic mosaic symptoms were observed on the 4th leaves 10 days after inoculation (dpi) with BSMV:γ or BSMV:Hv-SGT1. Photobleaching was observed on leaves infected with BSMV:PDS 15 dpi, whereas no detectable phenotype was observed in the mock-treated plants inoculated with 1×GKP buffer. Representative photographs were taken 15 days after virus inoculation. (B) The Hv-SGT1 gene was efficiently silenced in the BSMV: Hv-SGT1 inoculated leaves compared to the control inoculated with BSMV:γ, as indicated using qRT-PCR analysis. (C) More infected Bgt could developed into secondary hyphae in the BSMV: Hv-SGT1 inoculated leaves compared to the BSMV:γ infected leaves 10 days after inoculation with Bgt. (D) The rate of formation of secondary hyphae (SH) was higher in the Hv-SGT1 silenced leaves inoculated with BSMV:Hv-SGT1 compared with that in BSMV:γ-infected leaves at 5, 7, and 10 days after inoculation with Bgt. (E) Levels of NADPHOX, APX, NPR1, PR5, OPDA, COI1, ERF1, PR10, PR2, PR3, GSTand GPOXin Hv-SGT1 silenced leaves. * p < 0.05, ** p < 0.01 compared to the BSMV:γ-infected leaves.

Mentions: First, checking the efficiency of the VIGS system revealed that mild chlorotic mosaic symptoms were observed on the fourth leaves inoculated with BSMV at 10 days after inoculation. Photobleaching was observed on leaves infected with BSMV:PDS at 15 days, whereas no obvious changes were observed for mock leaves inoculated with 1×GKP buffer (Figure 5A). The level of expression of Hv-SGT1 was checked by qRT-PCR and found to decrease dramatically in H. villosa plants inoculated with BSMV:Hv-SGT1 compared with the Bgt inoculation plants preciously infected with BSMV: γ (Figure 5B).


The Hv-SGT1 gene from Haynaldia villosa contributes to resistances towards both biotrophic and hemi-biotrophic pathogens in common wheat (Triticum aestivum L.).

Xing L, Qian C, Cao A, Li Y, Jiang Z, Li M, Jin X, Hu J, Zhang Y, Wang X, Chen P - PLoS ONE (2013)

Functional analysis of the Hv-SGT1 gene by BSMV-induced gene silencing.(A) Following infection of H. villosa with the virus, chlorotic mosaic symptoms were observed on the 4th leaves 10 days after inoculation (dpi) with BSMV:γ or BSMV:Hv-SGT1. Photobleaching was observed on leaves infected with BSMV:PDS 15 dpi, whereas no detectable phenotype was observed in the mock-treated plants inoculated with 1×GKP buffer. Representative photographs were taken 15 days after virus inoculation. (B) The Hv-SGT1 gene was efficiently silenced in the BSMV: Hv-SGT1 inoculated leaves compared to the control inoculated with BSMV:γ, as indicated using qRT-PCR analysis. (C) More infected Bgt could developed into secondary hyphae in the BSMV: Hv-SGT1 inoculated leaves compared to the BSMV:γ infected leaves 10 days after inoculation with Bgt. (D) The rate of formation of secondary hyphae (SH) was higher in the Hv-SGT1 silenced leaves inoculated with BSMV:Hv-SGT1 compared with that in BSMV:γ-infected leaves at 5, 7, and 10 days after inoculation with Bgt. (E) Levels of NADPHOX, APX, NPR1, PR5, OPDA, COI1, ERF1, PR10, PR2, PR3, GSTand GPOXin Hv-SGT1 silenced leaves. * p < 0.05, ** p < 0.01 compared to the BSMV:γ-infected leaves.
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pone-0072571-g005: Functional analysis of the Hv-SGT1 gene by BSMV-induced gene silencing.(A) Following infection of H. villosa with the virus, chlorotic mosaic symptoms were observed on the 4th leaves 10 days after inoculation (dpi) with BSMV:γ or BSMV:Hv-SGT1. Photobleaching was observed on leaves infected with BSMV:PDS 15 dpi, whereas no detectable phenotype was observed in the mock-treated plants inoculated with 1×GKP buffer. Representative photographs were taken 15 days after virus inoculation. (B) The Hv-SGT1 gene was efficiently silenced in the BSMV: Hv-SGT1 inoculated leaves compared to the control inoculated with BSMV:γ, as indicated using qRT-PCR analysis. (C) More infected Bgt could developed into secondary hyphae in the BSMV: Hv-SGT1 inoculated leaves compared to the BSMV:γ infected leaves 10 days after inoculation with Bgt. (D) The rate of formation of secondary hyphae (SH) was higher in the Hv-SGT1 silenced leaves inoculated with BSMV:Hv-SGT1 compared with that in BSMV:γ-infected leaves at 5, 7, and 10 days after inoculation with Bgt. (E) Levels of NADPHOX, APX, NPR1, PR5, OPDA, COI1, ERF1, PR10, PR2, PR3, GSTand GPOXin Hv-SGT1 silenced leaves. * p < 0.05, ** p < 0.01 compared to the BSMV:γ-infected leaves.
Mentions: First, checking the efficiency of the VIGS system revealed that mild chlorotic mosaic symptoms were observed on the fourth leaves inoculated with BSMV at 10 days after inoculation. Photobleaching was observed on leaves infected with BSMV:PDS at 15 days, whereas no obvious changes were observed for mock leaves inoculated with 1×GKP buffer (Figure 5A). The level of expression of Hv-SGT1 was checked by qRT-PCR and found to decrease dramatically in H. villosa plants inoculated with BSMV:Hv-SGT1 compared with the Bgt inoculation plants preciously infected with BSMV: γ (Figure 5B).

Bottom Line: The demonstration that silencing of Hv-SGT1 substantially reduced resistance to Bgt indicated that Hv-SGT1 was an essential component of disease resistance in H. villosa.Therefore, the involvement of Hv-SGT1 in H2O2 production correlates with the hypersensitive response and jasmonic acid signaling.Our novel demonstration that wheat with over-expressed Hv-SGT1 showed enhanced resistance to both powdery mildew and FHB suggests that it could served as a transgenic genetic resource in wheat breeding for multiple disease resistance.

View Article: PubMed Central - PubMed

Affiliation: The National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
The SGT1 protein is essential for R protein-mediated and PAMPs-triggered resistance in many plant species. Here we reported the isolation and characterization of the Hv-SGT1 gene from Haynaldiavillosa (2n = 14, VV). Analysis of the subcellular location of Hv-SGT1 by transient expression of a fusion to GFP indicated its presence in the cytoplasm and nucleus. Levels of Hv-SGT1 transcripts were increased by inoculation with either the biotrophic pathogen Blumeriagraminis DC. f. Sp. tritici (Bgt) or the hemi-biotrophic pathogen Fusariumgraminearum (Fg). Levels of Hv-SGT1 showed substantial increase following treatment with H2O2 and methyl jasmonate (MeJA), only slightly induced following exposure to ethephon or abscisic acid, but not changed following exposure to salicylic acid. The demonstration that silencing of Hv-SGT1 substantially reduced resistance to Bgt indicated that Hv-SGT1 was an essential component of disease resistance in H. villosa. The over-expression of Hv-SGT1 in Yangmai 158 enhanced resistance to powdery mildew, and this correlated with increased levels of whole-cell reactive oxygen intermediates at the sites of penetration by the pathogens. Compared with wild-type plants, the expression levels of genes related to the H2O2 and JA signaling pathways were lower in the Hv-SGT1 silenced plants and higher in the Hv-SGT1 over-expressing plants. Therefore, the involvement of Hv-SGT1 in H2O2 production correlates with the hypersensitive response and jasmonic acid signaling. Our novel demonstration that wheat with over-expressed Hv-SGT1 showed enhanced resistance to both powdery mildew and FHB suggests that it could served as a transgenic genetic resource in wheat breeding for multiple disease resistance.

Show MeSH
Related in: MedlinePlus