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Characterization of a Novel Cotton Subtilase Gene GbSBT1 in Response to Extracellular Stimulations and Its Role in Verticillium Resistance.

Duan X, Zhang Z, Wang J, Zuo K - PLoS ONE (2016)

Bottom Line: Verticillium pathogens secrete various disease-causing effectors in cotton.Moreover, the GbSBT1 protein is mainly localized in the cell membrane and moves into the cytoplasm following jasmonic acid and ethylene treatments.In summary, GbSBT1 recognizes the effector PHB protein secreted from V. dahliae and is involved in Verticillium-induced resistance in cotton.

View Article: PubMed Central - PubMed

Affiliation: Plant Biotechnology Research Center, School of Agriculture and Life Sciences, Shanghai Jiao Tong University, Shanghai, China.

ABSTRACT
Verticillium wilt is a disastrous vascular disease in plants caused by Verticillium dahliae. Verticillium pathogens secrete various disease-causing effectors in cotton. This study identified a subtilase gene GbSBT1 from Gossypium babardense and investigated the roles against V. dahliae infection. GbSBT1 gene expression is responsive to V. dahliae defense signals, jasmonic acid, and ethylene treatments. Moreover, the GbSBT1 protein is mainly localized in the cell membrane and moves into the cytoplasm following jasmonic acid and ethylene treatments. Silencing GbSBT1 gene expression through virus-induced GbSBT1 gene silencing reduced the tolerance of Pima-90 (resistant genotype), but not facilitated the infection process of V. dahliae in Coker-312 (sensitive genotype). Moreover, the ectopically expressed GbSBT1 gene enhanced the resistance of Arabidopsis to Fusarium oxysporum and V. dahliae infection and activated the expression levels of defense-related genes. Furthermore, pull-down, yeast two-hybrid assay, and BiFC analysis revealed that GbSBT1 interacts with a prohibitin (PHB)-like protein expressed in V. dahliae pathogens during infection. In summary, GbSBT1 recognizes the effector PHB protein secreted from V. dahliae and is involved in Verticillium-induced resistance in cotton.

No MeSH data available.


Related in: MedlinePlus

Extracellular localization of the GbSBT1 protein in tobacco leaves determined through confocal microscopy.(A) Localization of GbSBT1-YFP in tobacco leaves; localization of GbSBT1 (without signal peptide)-YFP in tobacco leaves; localization of GbSBT1-YFP in tobacco leaves under JA treatment; localization of GbSBT1-YFP in tobacco leaves under ethylene treatment. Arrows indicate the difference in protein localization between normal and phytohormone treatments; (B) GbSBT1 protein co-localized with plasma membrane integral protein PIP1 on the cell membrane using protoplast and tobacco lead epidermal cells as the protein expressing materials. Upper panels: protoplast; lower panels: tobacco leaf epidermal cells.
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pone.0153988.g004: Extracellular localization of the GbSBT1 protein in tobacco leaves determined through confocal microscopy.(A) Localization of GbSBT1-YFP in tobacco leaves; localization of GbSBT1 (without signal peptide)-YFP in tobacco leaves; localization of GbSBT1-YFP in tobacco leaves under JA treatment; localization of GbSBT1-YFP in tobacco leaves under ethylene treatment. Arrows indicate the difference in protein localization between normal and phytohormone treatments; (B) GbSBT1 protein co-localized with plasma membrane integral protein PIP1 on the cell membrane using protoplast and tobacco lead epidermal cells as the protein expressing materials. Upper panels: protoplast; lower panels: tobacco leaf epidermal cells.

Mentions: To analyze GbSBT1 subcellular localization, we expressed the ORF of the GbSBT1 gene fused with eYFP in tobacco leaves. As shown in Fig 4A, GbSBT1-eYFP fluorescence signals were detected in the plasma membrane. When magnifying the accumulation area, GbSBT1-eYFP signals were unevenly distributed in the cell membrane (Fig 4A), which provides the conduits for the exchange of informational molecules that are central to cell growth and defense response in plants. When GbSBT1 without the signal peptide was expressed in tobacco leaves, GbSBT1 (no SP)-YFP signals were uniformly distributed in the plasma membrane. The co-localization signals with plasma membrane integral protein PIP1-mcherry further demonstrate that GbSBT1 mostly targets the cell membrane (Fig 4B). Overall, GbSBT1 is mainly localized in the cell membrane. GbSBT1 extracellular localization may be linked to the acceptance of external signals.


Characterization of a Novel Cotton Subtilase Gene GbSBT1 in Response to Extracellular Stimulations and Its Role in Verticillium Resistance.

Duan X, Zhang Z, Wang J, Zuo K - PLoS ONE (2016)

Extracellular localization of the GbSBT1 protein in tobacco leaves determined through confocal microscopy.(A) Localization of GbSBT1-YFP in tobacco leaves; localization of GbSBT1 (without signal peptide)-YFP in tobacco leaves; localization of GbSBT1-YFP in tobacco leaves under JA treatment; localization of GbSBT1-YFP in tobacco leaves under ethylene treatment. Arrows indicate the difference in protein localization between normal and phytohormone treatments; (B) GbSBT1 protein co-localized with plasma membrane integral protein PIP1 on the cell membrane using protoplast and tobacco lead epidermal cells as the protein expressing materials. Upper panels: protoplast; lower panels: tobacco leaf epidermal cells.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835097&req=5

pone.0153988.g004: Extracellular localization of the GbSBT1 protein in tobacco leaves determined through confocal microscopy.(A) Localization of GbSBT1-YFP in tobacco leaves; localization of GbSBT1 (without signal peptide)-YFP in tobacco leaves; localization of GbSBT1-YFP in tobacco leaves under JA treatment; localization of GbSBT1-YFP in tobacco leaves under ethylene treatment. Arrows indicate the difference in protein localization between normal and phytohormone treatments; (B) GbSBT1 protein co-localized with plasma membrane integral protein PIP1 on the cell membrane using protoplast and tobacco lead epidermal cells as the protein expressing materials. Upper panels: protoplast; lower panels: tobacco leaf epidermal cells.
Mentions: To analyze GbSBT1 subcellular localization, we expressed the ORF of the GbSBT1 gene fused with eYFP in tobacco leaves. As shown in Fig 4A, GbSBT1-eYFP fluorescence signals were detected in the plasma membrane. When magnifying the accumulation area, GbSBT1-eYFP signals were unevenly distributed in the cell membrane (Fig 4A), which provides the conduits for the exchange of informational molecules that are central to cell growth and defense response in plants. When GbSBT1 without the signal peptide was expressed in tobacco leaves, GbSBT1 (no SP)-YFP signals were uniformly distributed in the plasma membrane. The co-localization signals with plasma membrane integral protein PIP1-mcherry further demonstrate that GbSBT1 mostly targets the cell membrane (Fig 4B). Overall, GbSBT1 is mainly localized in the cell membrane. GbSBT1 extracellular localization may be linked to the acceptance of external signals.

Bottom Line: Verticillium pathogens secrete various disease-causing effectors in cotton.Moreover, the GbSBT1 protein is mainly localized in the cell membrane and moves into the cytoplasm following jasmonic acid and ethylene treatments.In summary, GbSBT1 recognizes the effector PHB protein secreted from V. dahliae and is involved in Verticillium-induced resistance in cotton.

View Article: PubMed Central - PubMed

Affiliation: Plant Biotechnology Research Center, School of Agriculture and Life Sciences, Shanghai Jiao Tong University, Shanghai, China.

ABSTRACT
Verticillium wilt is a disastrous vascular disease in plants caused by Verticillium dahliae. Verticillium pathogens secrete various disease-causing effectors in cotton. This study identified a subtilase gene GbSBT1 from Gossypium babardense and investigated the roles against V. dahliae infection. GbSBT1 gene expression is responsive to V. dahliae defense signals, jasmonic acid, and ethylene treatments. Moreover, the GbSBT1 protein is mainly localized in the cell membrane and moves into the cytoplasm following jasmonic acid and ethylene treatments. Silencing GbSBT1 gene expression through virus-induced GbSBT1 gene silencing reduced the tolerance of Pima-90 (resistant genotype), but not facilitated the infection process of V. dahliae in Coker-312 (sensitive genotype). Moreover, the ectopically expressed GbSBT1 gene enhanced the resistance of Arabidopsis to Fusarium oxysporum and V. dahliae infection and activated the expression levels of defense-related genes. Furthermore, pull-down, yeast two-hybrid assay, and BiFC analysis revealed that GbSBT1 interacts with a prohibitin (PHB)-like protein expressed in V. dahliae pathogens during infection. In summary, GbSBT1 recognizes the effector PHB protein secreted from V. dahliae and is involved in Verticillium-induced resistance in cotton.

No MeSH data available.


Related in: MedlinePlus