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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

Interaction between GbSBT1 and prohibitin secreted by Verticillium dahliae.(A) The yeast cells co-expressing GbSBT1 and benzodiazepine receptor family protein or peptidyl-tRNA hydrolase could not grow on the selective medium SD/-T-L-H, whereas the yeast cells co-transformed with GbSBT1 and prohibitin could grow on the selective medium SD/-T-L-H; (B-C) BiFC of epidermal cells co-expressing split YFP fusions of GbSBT1 and prohibitin or empty vector controls. Combinations of N- and C-terminal YFP fragments (Yn and Yc, respectively) were infiltrated as vector controls or fused to the N terminus of GbSBT1 and prohibitin as follows: B: GbSBT1-Yc and prohibitin -Yn, Scale bar: 25 μm; C: vector-Yc and vector-Yn, Scale bar: 10 μm. The interaction and co-localization were observed at the plasma membrane. Right is the corresponding bright-field
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pone.0153988.g010: Interaction between GbSBT1 and prohibitin secreted by Verticillium dahliae.(A) The yeast cells co-expressing GbSBT1 and benzodiazepine receptor family protein or peptidyl-tRNA hydrolase could not grow on the selective medium SD/-T-L-H, whereas the yeast cells co-transformed with GbSBT1 and prohibitin could grow on the selective medium SD/-T-L-H; (B-C) BiFC of epidermal cells co-expressing split YFP fusions of GbSBT1 and prohibitin or empty vector controls. Combinations of N- and C-terminal YFP fragments (Yn and Yc, respectively) were infiltrated as vector controls or fused to the N terminus of GbSBT1 and prohibitin as follows: B: GbSBT1-Yc and prohibitin -Yn, Scale bar: 25 μm; C: vector-Yc and vector-Yn, Scale bar: 10 μm. The interaction and co-localization were observed at the plasma membrane. Right is the corresponding bright-field

Mentions: To further confirm which protein directly interacts with GbSBT1 during V. dahliae ingression into cotton, three proteins (based on the number of detected peptides in pull-down assay) were tested their interactions with GbSBT1 in yeast-two-hybridization. As shown in Fig 10A, GbSBT1 did not interact with the benzodiazepine receptor family protein and peptidyl-tRNA hydrolase, whereas the yeast cells co-transformed with GbSBT1 and PHB grew on the selective medium SD/-T-L-H. This result indicates that GbSBT1 can directly bind to PHB.


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)

Interaction between GbSBT1 and prohibitin secreted by Verticillium dahliae.(A) The yeast cells co-expressing GbSBT1 and benzodiazepine receptor family protein or peptidyl-tRNA hydrolase could not grow on the selective medium SD/-T-L-H, whereas the yeast cells co-transformed with GbSBT1 and prohibitin could grow on the selective medium SD/-T-L-H; (B-C) BiFC of epidermal cells co-expressing split YFP fusions of GbSBT1 and prohibitin or empty vector controls. Combinations of N- and C-terminal YFP fragments (Yn and Yc, respectively) were infiltrated as vector controls or fused to the N terminus of GbSBT1 and prohibitin as follows: B: GbSBT1-Yc and prohibitin -Yn, Scale bar: 25 μm; C: vector-Yc and vector-Yn, Scale bar: 10 μm. The interaction and co-localization were observed at the plasma membrane. Right is the corresponding bright-field
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153988.g010: Interaction between GbSBT1 and prohibitin secreted by Verticillium dahliae.(A) The yeast cells co-expressing GbSBT1 and benzodiazepine receptor family protein or peptidyl-tRNA hydrolase could not grow on the selective medium SD/-T-L-H, whereas the yeast cells co-transformed with GbSBT1 and prohibitin could grow on the selective medium SD/-T-L-H; (B-C) BiFC of epidermal cells co-expressing split YFP fusions of GbSBT1 and prohibitin or empty vector controls. Combinations of N- and C-terminal YFP fragments (Yn and Yc, respectively) were infiltrated as vector controls or fused to the N terminus of GbSBT1 and prohibitin as follows: B: GbSBT1-Yc and prohibitin -Yn, Scale bar: 25 μm; C: vector-Yc and vector-Yn, Scale bar: 10 μm. The interaction and co-localization were observed at the plasma membrane. Right is the corresponding bright-field
Mentions: To further confirm which protein directly interacts with GbSBT1 during V. dahliae ingression into cotton, three proteins (based on the number of detected peptides in pull-down assay) were tested their interactions with GbSBT1 in yeast-two-hybridization. As shown in Fig 10A, GbSBT1 did not interact with the benzodiazepine receptor family protein and peptidyl-tRNA hydrolase, whereas the yeast cells co-transformed with GbSBT1 and PHB grew on the selective medium SD/-T-L-H. This result indicates that GbSBT1 can directly bind to PHB.

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