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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

Phylogenetic analysis of GbSBT1 including the subtilases of different plant species.Nineteen of the complete amino acid sequences of subtilases were used to generate the neighbor–joining tree, and the numbers next to each node provide bootstrap values from 1000 replicates. The GbSBT1 gene is marked by “◆.” GbSBT1 (KT336228, Gossypium barbadense), SBT-like Picea abies (BAA13135, Picea sitchensis), AtSBT5.2 (NP_564107, Arabidopsis thaliana), SBT-like1 Theobroma cacao (XP_007017870, T. cacao), SBT-like2 T. cacao (XP_007017871, T. cacao), SBT Arabidopsis lyrata (XP_002893091, A. lyrata), SBT-like1 A. thaliana (AAM65424, A. thaliana), SBT-like1 Morus notabilis (EXB60669, M. notabilis), SBT1 Medicago truncatula (XP_003603196, M. truncatula), SBT2 M. truncatula (XP_003603807, M. truncatula), SBT1 Glycine max (NP_001236511, G. max), SBT2 G. max (NP_001238252, G. max), SBT-like G. max (AAK53589, G. max), SBT-like2 A. thaliana (NP_564106, A. thaliana), SBT-like Aegilops tauschii (EMT32761, A. tauschii), SBT-like3 T. cacao (XP_007028364, T. cacao), SBT-like2 M. notabilis (EXB44295, M. notabilis), SBT-like3 M. notabilis (EXB44294, M. notabilis), SBT-like3 A. thaliana (NP_565309, A. thaliana).
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pone.0153988.g002: Phylogenetic analysis of GbSBT1 including the subtilases of different plant species.Nineteen of the complete amino acid sequences of subtilases were used to generate the neighbor–joining tree, and the numbers next to each node provide bootstrap values from 1000 replicates. The GbSBT1 gene is marked by “◆.” GbSBT1 (KT336228, Gossypium barbadense), SBT-like Picea abies (BAA13135, Picea sitchensis), AtSBT5.2 (NP_564107, Arabidopsis thaliana), SBT-like1 Theobroma cacao (XP_007017870, T. cacao), SBT-like2 T. cacao (XP_007017871, T. cacao), SBT Arabidopsis lyrata (XP_002893091, A. lyrata), SBT-like1 A. thaliana (AAM65424, A. thaliana), SBT-like1 Morus notabilis (EXB60669, M. notabilis), SBT1 Medicago truncatula (XP_003603196, M. truncatula), SBT2 M. truncatula (XP_003603807, M. truncatula), SBT1 Glycine max (NP_001236511, G. max), SBT2 G. max (NP_001238252, G. max), SBT-like G. max (AAK53589, G. max), SBT-like2 A. thaliana (NP_564106, A. thaliana), SBT-like Aegilops tauschii (EMT32761, A. tauschii), SBT-like3 T. cacao (XP_007028364, T. cacao), SBT-like2 M. notabilis (EXB44295, M. notabilis), SBT-like3 M. notabilis (EXB44294, M. notabilis), SBT-like3 A. thaliana (NP_565309, A. thaliana).

Mentions: The GbSBT1 gene encodes a protein containing 769 aa, corresponding to a molecular mass of 81.78 kDa and a pI of 6.34. The GbSBT1 protein contains a putative signal peptide, an inhibitor domain, five conserved active sites (Ser-544, Asp-145, and His-223), and two catalytic triads of serine protease (Fig 1). A neighbor-joining phylogenetic tree of 19 subtilases indicated that that GbSBT1 is closely related to AtSBT5.2 (Fig 2). In Arabidopsis, AtSBT5.2 exhibits oxido-reductase activity and responds to an external biotic stimulus [10], indicating that the GbSBT1 gene may function in biotic stress response.


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)

Phylogenetic analysis of GbSBT1 including the subtilases of different plant species.Nineteen of the complete amino acid sequences of subtilases were used to generate the neighbor–joining tree, and the numbers next to each node provide bootstrap values from 1000 replicates. The GbSBT1 gene is marked by “◆.” GbSBT1 (KT336228, Gossypium barbadense), SBT-like Picea abies (BAA13135, Picea sitchensis), AtSBT5.2 (NP_564107, Arabidopsis thaliana), SBT-like1 Theobroma cacao (XP_007017870, T. cacao), SBT-like2 T. cacao (XP_007017871, T. cacao), SBT Arabidopsis lyrata (XP_002893091, A. lyrata), SBT-like1 A. thaliana (AAM65424, A. thaliana), SBT-like1 Morus notabilis (EXB60669, M. notabilis), SBT1 Medicago truncatula (XP_003603196, M. truncatula), SBT2 M. truncatula (XP_003603807, M. truncatula), SBT1 Glycine max (NP_001236511, G. max), SBT2 G. max (NP_001238252, G. max), SBT-like G. max (AAK53589, G. max), SBT-like2 A. thaliana (NP_564106, A. thaliana), SBT-like Aegilops tauschii (EMT32761, A. tauschii), SBT-like3 T. cacao (XP_007028364, T. cacao), SBT-like2 M. notabilis (EXB44295, M. notabilis), SBT-like3 M. notabilis (EXB44294, M. notabilis), SBT-like3 A. thaliana (NP_565309, A. thaliana).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153988.g002: Phylogenetic analysis of GbSBT1 including the subtilases of different plant species.Nineteen of the complete amino acid sequences of subtilases were used to generate the neighbor–joining tree, and the numbers next to each node provide bootstrap values from 1000 replicates. The GbSBT1 gene is marked by “◆.” GbSBT1 (KT336228, Gossypium barbadense), SBT-like Picea abies (BAA13135, Picea sitchensis), AtSBT5.2 (NP_564107, Arabidopsis thaliana), SBT-like1 Theobroma cacao (XP_007017870, T. cacao), SBT-like2 T. cacao (XP_007017871, T. cacao), SBT Arabidopsis lyrata (XP_002893091, A. lyrata), SBT-like1 A. thaliana (AAM65424, A. thaliana), SBT-like1 Morus notabilis (EXB60669, M. notabilis), SBT1 Medicago truncatula (XP_003603196, M. truncatula), SBT2 M. truncatula (XP_003603807, M. truncatula), SBT1 Glycine max (NP_001236511, G. max), SBT2 G. max (NP_001238252, G. max), SBT-like G. max (AAK53589, G. max), SBT-like2 A. thaliana (NP_564106, A. thaliana), SBT-like Aegilops tauschii (EMT32761, A. tauschii), SBT-like3 T. cacao (XP_007028364, T. cacao), SBT-like2 M. notabilis (EXB44295, M. notabilis), SBT-like3 M. notabilis (EXB44294, M. notabilis), SBT-like3 A. thaliana (NP_565309, A. thaliana).
Mentions: The GbSBT1 gene encodes a protein containing 769 aa, corresponding to a molecular mass of 81.78 kDa and a pI of 6.34. The GbSBT1 protein contains a putative signal peptide, an inhibitor domain, five conserved active sites (Ser-544, Asp-145, and His-223), and two catalytic triads of serine protease (Fig 1). A neighbor-joining phylogenetic tree of 19 subtilases indicated that that GbSBT1 is closely related to AtSBT5.2 (Fig 2). In Arabidopsis, AtSBT5.2 exhibits oxido-reductase activity and responds to an external biotic stimulus [10], indicating that the GbSBT1 gene may function in biotic stress response.

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