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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.


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qPCR analysis of gene expression in WT and transgenic GbSBT1 lines after F. oxysporum infection.Gene expression levels were normalized to the constitutive ACT2 gene expression. Data represent the mean of gene expression levels with SD at different infection stages; n = 3 replicates. dpi: days post-inoculation. Error bars represent the standard deviation for three biological experiments, and three technical replicates were analyzed. Statistical significance was determined using two-tailed unpaired Student’s t-tests, and P values <0.05 were considered statistically significant (** for P values <0.01).
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pone.0153988.g009: qPCR analysis of gene expression in WT and transgenic GbSBT1 lines after F. oxysporum infection.Gene expression levels were normalized to the constitutive ACT2 gene expression. Data represent the mean of gene expression levels with SD at different infection stages; n = 3 replicates. dpi: days post-inoculation. Error bars represent the standard deviation for three biological experiments, and three technical replicates were analyzed. Statistical significance was determined using two-tailed unpaired Student’s t-tests, and P values <0.05 were considered statistically significant (** for P values <0.01).

Mentions: Given their consistent resistance effects on GbSBT1-overexpressing Arabidopsis plants and VIGS cotton plants, the transcripts of the genes involved in JA and immune transduction were analyzed in Arabidopsis. The expression levels of PR-2 and PDF1.2 were upregulated in the overexpressing lines following F. oxysporum inoculation (Fig 9). PR-2 was induced, peaked at 4 dpi, and then abruptly decreased. The highest level of PR-2 expression was nearly 10 times that of the WT plants at 4 dpi. PDF1.2 induction was peaked at 4 dpi; the highest expression of this gene was approximately 98.8 times as that of the control plants. The increased expression of PRs in GbSBT1 OEX lines contributed to the resistant phenotype.


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)

qPCR analysis of gene expression in WT and transgenic GbSBT1 lines after F. oxysporum infection.Gene expression levels were normalized to the constitutive ACT2 gene expression. Data represent the mean of gene expression levels with SD at different infection stages; n = 3 replicates. dpi: days post-inoculation. Error bars represent the standard deviation for three biological experiments, and three technical replicates were analyzed. Statistical significance was determined using two-tailed unpaired Student’s t-tests, and P values <0.05 were considered statistically significant (** for P values <0.01).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0153988.g009: qPCR analysis of gene expression in WT and transgenic GbSBT1 lines after F. oxysporum infection.Gene expression levels were normalized to the constitutive ACT2 gene expression. Data represent the mean of gene expression levels with SD at different infection stages; n = 3 replicates. dpi: days post-inoculation. Error bars represent the standard deviation for three biological experiments, and three technical replicates were analyzed. Statistical significance was determined using two-tailed unpaired Student’s t-tests, and P values <0.05 were considered statistically significant (** for P values <0.01).
Mentions: Given their consistent resistance effects on GbSBT1-overexpressing Arabidopsis plants and VIGS cotton plants, the transcripts of the genes involved in JA and immune transduction were analyzed in Arabidopsis. The expression levels of PR-2 and PDF1.2 were upregulated in the overexpressing lines following F. oxysporum inoculation (Fig 9). PR-2 was induced, peaked at 4 dpi, and then abruptly decreased. The highest level of PR-2 expression was nearly 10 times that of the WT plants at 4 dpi. PDF1.2 induction was peaked at 4 dpi; the highest expression of this gene was approximately 98.8 times as that of the control plants. The increased expression of PRs in GbSBT1 OEX lines contributed to the resistant phenotype.

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