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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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Related in: MedlinePlus

Characterization of the transgenic wheat of Hv-SGT1.(A) PCR and PCR-Southern blot of four transgenic lines that over-expressing Hv-SGT1 (OX), and the non-transformed control Yangmai 158. The plasmid pBI220.6-Hv-SGT1 and Yangmai 158 were used as the positive and negative controls, respectively. (B) qRT-PCR of the expression of Hv-SGT1 in the four transgenic lines and Yangmai 158. ** p < 0.01 compared with the control. (C) Reduced disease symptoms in transgenic plants. Seedling resistance of transgenic line or wild-type plants was assessed following in vitro infection with the native pathogen population (Sumai 3). (D) Microscopic observation of Bgt hyphae spreading after DioC6 staining of the transgenic plants and Yangmai 158. (E) Quantitative comparisons of the percentage of infection sites with secondary hyphae (SH), the average number of hyphal branches and average hyphal length emerging on the leaves of infection sites. Means (± SE) were calculated using the measurements from five seedlings, and at least 30 infection sites for each seedling. Significance was determined according to paired sample t-test method (b indicates P < 0.05).
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pone-0072571-g006: Characterization of the transgenic wheat of Hv-SGT1.(A) PCR and PCR-Southern blot of four transgenic lines that over-expressing Hv-SGT1 (OX), and the non-transformed control Yangmai 158. The plasmid pBI220.6-Hv-SGT1 and Yangmai 158 were used as the positive and negative controls, respectively. (B) qRT-PCR of the expression of Hv-SGT1 in the four transgenic lines and Yangmai 158. ** p < 0.01 compared with the control. (C) Reduced disease symptoms in transgenic plants. Seedling resistance of transgenic line or wild-type plants was assessed following in vitro infection with the native pathogen population (Sumai 3). (D) Microscopic observation of Bgt hyphae spreading after DioC6 staining of the transgenic plants and Yangmai 158. (E) Quantitative comparisons of the percentage of infection sites with secondary hyphae (SH), the average number of hyphal branches and average hyphal length emerging on the leaves of infection sites. Means (± SE) were calculated using the measurements from five seedlings, and at least 30 infection sites for each seedling. Significance was determined according to paired sample t-test method (b indicates P < 0.05).

Mentions: Although it has been well documented that SGT1 regulated defense responses triggered by various pathogens, less research had investigated its effectiveness to both biothophic and hemi-biothophic pathogens. Transgenic Yangmai 158 plants that over-express Hv-SGT1 under the regulation of the CAMV35S promoter were obtained by using the particle bombardment-mediated transformation approach. Overall, 52 independent T0-generation transgenic plants were identified after analysis of 432 regenerants using PCR amplification. Four of these lines (OX-313, OX-322, OX-323, OX-330) were characterized by further Southern blot of the PCR products (Figure 6A). qRT-PCR of the identified four lines indicated that expression of Hv-SGT1 in the transgenic plants was significantly increased up to 7.5-fold compared with that in the non-transformed Yangmai 158 (Figure 6B). The progenitors derived from these four positive transgenic plants were analyzed to assess their resistances to powdery mildew.


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)

Characterization of the transgenic wheat of Hv-SGT1.(A) PCR and PCR-Southern blot of four transgenic lines that over-expressing Hv-SGT1 (OX), and the non-transformed control Yangmai 158. The plasmid pBI220.6-Hv-SGT1 and Yangmai 158 were used as the positive and negative controls, respectively. (B) qRT-PCR of the expression of Hv-SGT1 in the four transgenic lines and Yangmai 158. ** p < 0.01 compared with the control. (C) Reduced disease symptoms in transgenic plants. Seedling resistance of transgenic line or wild-type plants was assessed following in vitro infection with the native pathogen population (Sumai 3). (D) Microscopic observation of Bgt hyphae spreading after DioC6 staining of the transgenic plants and Yangmai 158. (E) Quantitative comparisons of the percentage of infection sites with secondary hyphae (SH), the average number of hyphal branches and average hyphal length emerging on the leaves of infection sites. Means (± SE) were calculated using the measurements from five seedlings, and at least 30 infection sites for each seedling. Significance was determined according to paired sample t-test method (b indicates P < 0.05).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760960&req=5

pone-0072571-g006: Characterization of the transgenic wheat of Hv-SGT1.(A) PCR and PCR-Southern blot of four transgenic lines that over-expressing Hv-SGT1 (OX), and the non-transformed control Yangmai 158. The plasmid pBI220.6-Hv-SGT1 and Yangmai 158 were used as the positive and negative controls, respectively. (B) qRT-PCR of the expression of Hv-SGT1 in the four transgenic lines and Yangmai 158. ** p < 0.01 compared with the control. (C) Reduced disease symptoms in transgenic plants. Seedling resistance of transgenic line or wild-type plants was assessed following in vitro infection with the native pathogen population (Sumai 3). (D) Microscopic observation of Bgt hyphae spreading after DioC6 staining of the transgenic plants and Yangmai 158. (E) Quantitative comparisons of the percentage of infection sites with secondary hyphae (SH), the average number of hyphal branches and average hyphal length emerging on the leaves of infection sites. Means (± SE) were calculated using the measurements from five seedlings, and at least 30 infection sites for each seedling. Significance was determined according to paired sample t-test method (b indicates P < 0.05).
Mentions: Although it has been well documented that SGT1 regulated defense responses triggered by various pathogens, less research had investigated its effectiveness to both biothophic and hemi-biothophic pathogens. Transgenic Yangmai 158 plants that over-express Hv-SGT1 under the regulation of the CAMV35S promoter were obtained by using the particle bombardment-mediated transformation approach. Overall, 52 independent T0-generation transgenic plants were identified after analysis of 432 regenerants using PCR amplification. Four of these lines (OX-313, OX-322, OX-323, OX-330) were characterized by further Southern blot of the PCR products (Figure 6A). qRT-PCR of the identified four lines indicated that expression of Hv-SGT1 in the transgenic plants was significantly increased up to 7.5-fold compared with that in the non-transformed Yangmai 158 (Figure 6B). The progenitors derived from these four positive transgenic plants were analyzed to assess their resistances to powdery mildew.

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