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Overexpression of the soybean GmERF3 gene, an AP2/ERF type transcription factor for increased tolerances to salt, drought, and diseases in transgenic tobacco.

Zhang G, Chen M, Li L, Xu Z, Chen X, Guo J, Ma Y - J. Exp. Bot. (2009)

Bottom Line: The GmERF3 protein fused to the GAL4 DNA-binding domain to activate transcription of reporter genes in yeast.Furthermore, overexpression of GmERF3 in transgenic tobacco led to higher levels of free proline and soluble carbohydrates compared to wild-type plants under drought conditions.The overall results suggested that GmERF3 as an AP2/ERF transcription factor may play dual roles in response to biotic and abiotic stresses in plants.

View Article: PubMed Central - PubMed

Affiliation: The National Key Facility for Crop Genetic Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
A new member of the AP2/ERF transcription factor family, GmERF3, was isolated from soybean. Sequence analysis showed that GmERF3 contained an AP2/ERF domain of 58 amino acids and two putative nuclear localization signal (NLS) domains. It belonged to a group IV protein in the ERF (ethylene response factor) subfamily as typified by a conserved N-terminal motif [MCGGAI(I/L)]. Expression of GmERF3 was induced by treatments with high salinity, drought, abscisic acid (ABA), salicylic acid (SA), jasmonic acid (JA), ethylene (ET), and soybean mosaic virus (SMV), whereas there was no significant GmERF3 mRNA accumulation under cold stress treatment. GmERF3 could bind to the GCC box and DRE/CRT element, and was targeted to the nucleus when transiently expressed in onion epidermal cells. The GmERF3 protein fused to the GAL4 DNA-binding domain to activate transcription of reporter genes in yeast. Ectopic expression of the GmERF3 gene in transgenic tobacco plants induced the expression of some PR genes and enhanced resistance against infection by Ralstonia solanacearum, Alternaria alternata, and tobacco mosaic virus (TMV), and gave tolerance to high salinity and dehydration stresses. Furthermore, overexpression of GmERF3 in transgenic tobacco led to higher levels of free proline and soluble carbohydrates compared to wild-type plants under drought conditions. The overall results suggested that GmERF3 as an AP2/ERF transcription factor may play dual roles in response to biotic and abiotic stresses in plants.

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Resistance analysis of GmERF3 transgenic tobacco plants. (A) Resistance of transgenic tobacco to the bacterial pathogen Ralstonia solanacearum. Fully expanded leaves of 7-week-old tobacco plants were syringe-infiltrated with 107 cfu ml−1 solution of R. solanacearum. Disease symptoms in wild-type and GmERF3 transgenic plants are shown in the upper panel; leaves in the upper line were inoculated with bacteria, and leaves below the lower line were mock-inoculated. The photograph was taken 7 d after inoculation. Infected leaves were collected and bacterial populations are shown in the lower panel. Values are means of three different experiments. Error bars indicate the SE. (B) Responses of transgenic tobacco to the fungal pathogen Alternaria alternata. Detached leaves were challenged with mycelia of A. alternata. The photograph was taken 7 d following inoculation. Disease symptoms are shown on the right. The average lesion area of each independent transgenic line (n=4) was calculated and their relative lesion areas are shown in columns after comparison with the average lesion area on wild-type tobacco. (C) Time-course for the systemic spread of TMV in wild-type and GmERF3 transgenic tobacco plants. The line in the middle of the upper panel indicates time. Notes above and below the line describe symptoms of wild-type and GmERF3 transgenic tobacco plants caused by TMV infection and spread, respectively. WT and G represent the wild-type and GmERF3 transgenic plants, respectively. Photographs were taken 12 d (right and middle of the lower panel) and 16 d (left of the lower panel) after inoculation. Arrows indicate TMV-inoculated leaves or mock-inoculated leaves. (This figure is available in colour at JXB online.)
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fig7: Resistance analysis of GmERF3 transgenic tobacco plants. (A) Resistance of transgenic tobacco to the bacterial pathogen Ralstonia solanacearum. Fully expanded leaves of 7-week-old tobacco plants were syringe-infiltrated with 107 cfu ml−1 solution of R. solanacearum. Disease symptoms in wild-type and GmERF3 transgenic plants are shown in the upper panel; leaves in the upper line were inoculated with bacteria, and leaves below the lower line were mock-inoculated. The photograph was taken 7 d after inoculation. Infected leaves were collected and bacterial populations are shown in the lower panel. Values are means of three different experiments. Error bars indicate the SE. (B) Responses of transgenic tobacco to the fungal pathogen Alternaria alternata. Detached leaves were challenged with mycelia of A. alternata. The photograph was taken 7 d following inoculation. Disease symptoms are shown on the right. The average lesion area of each independent transgenic line (n=4) was calculated and their relative lesion areas are shown in columns after comparison with the average lesion area on wild-type tobacco. (C) Time-course for the systemic spread of TMV in wild-type and GmERF3 transgenic tobacco plants. The line in the middle of the upper panel indicates time. Notes above and below the line describe symptoms of wild-type and GmERF3 transgenic tobacco plants caused by TMV infection and spread, respectively. WT and G represent the wild-type and GmERF3 transgenic plants, respectively. Photographs were taken 12 d (right and middle of the lower panel) and 16 d (left of the lower panel) after inoculation. Arrows indicate TMV-inoculated leaves or mock-inoculated leaves. (This figure is available in colour at JXB online.)

Mentions: To determine whether overexpression of GmERF3 enhanced resistance to pathogens in transgenic tobacco plants, the bacterial pathogen Ralstonia solanacearum was injected into leaves of wild-type and transgenic plants, and symptom development was subsequently monitored for 7 d. As shown in Fig. 7A, all transgenic lines exhibited significantly reduced disease lesions and leaf bacterial numbers compared to wild-type plants. Approximately 50% inhibition of bacterial growth was detected in 35S::GmERF3 tobacco plants during the 7-d infection period (P <0.01). Line G2 showed stronger resistance than other transgenic lines tested.


Overexpression of the soybean GmERF3 gene, an AP2/ERF type transcription factor for increased tolerances to salt, drought, and diseases in transgenic tobacco.

Zhang G, Chen M, Li L, Xu Z, Chen X, Guo J, Ma Y - J. Exp. Bot. (2009)

Resistance analysis of GmERF3 transgenic tobacco plants. (A) Resistance of transgenic tobacco to the bacterial pathogen Ralstonia solanacearum. Fully expanded leaves of 7-week-old tobacco plants were syringe-infiltrated with 107 cfu ml−1 solution of R. solanacearum. Disease symptoms in wild-type and GmERF3 transgenic plants are shown in the upper panel; leaves in the upper line were inoculated with bacteria, and leaves below the lower line were mock-inoculated. The photograph was taken 7 d after inoculation. Infected leaves were collected and bacterial populations are shown in the lower panel. Values are means of three different experiments. Error bars indicate the SE. (B) Responses of transgenic tobacco to the fungal pathogen Alternaria alternata. Detached leaves were challenged with mycelia of A. alternata. The photograph was taken 7 d following inoculation. Disease symptoms are shown on the right. The average lesion area of each independent transgenic line (n=4) was calculated and their relative lesion areas are shown in columns after comparison with the average lesion area on wild-type tobacco. (C) Time-course for the systemic spread of TMV in wild-type and GmERF3 transgenic tobacco plants. The line in the middle of the upper panel indicates time. Notes above and below the line describe symptoms of wild-type and GmERF3 transgenic tobacco plants caused by TMV infection and spread, respectively. WT and G represent the wild-type and GmERF3 transgenic plants, respectively. Photographs were taken 12 d (right and middle of the lower panel) and 16 d (left of the lower panel) after inoculation. Arrows indicate TMV-inoculated leaves or mock-inoculated leaves. (This figure is available in colour at JXB online.)
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2736888&req=5

fig7: Resistance analysis of GmERF3 transgenic tobacco plants. (A) Resistance of transgenic tobacco to the bacterial pathogen Ralstonia solanacearum. Fully expanded leaves of 7-week-old tobacco plants were syringe-infiltrated with 107 cfu ml−1 solution of R. solanacearum. Disease symptoms in wild-type and GmERF3 transgenic plants are shown in the upper panel; leaves in the upper line were inoculated with bacteria, and leaves below the lower line were mock-inoculated. The photograph was taken 7 d after inoculation. Infected leaves were collected and bacterial populations are shown in the lower panel. Values are means of three different experiments. Error bars indicate the SE. (B) Responses of transgenic tobacco to the fungal pathogen Alternaria alternata. Detached leaves were challenged with mycelia of A. alternata. The photograph was taken 7 d following inoculation. Disease symptoms are shown on the right. The average lesion area of each independent transgenic line (n=4) was calculated and their relative lesion areas are shown in columns after comparison with the average lesion area on wild-type tobacco. (C) Time-course for the systemic spread of TMV in wild-type and GmERF3 transgenic tobacco plants. The line in the middle of the upper panel indicates time. Notes above and below the line describe symptoms of wild-type and GmERF3 transgenic tobacco plants caused by TMV infection and spread, respectively. WT and G represent the wild-type and GmERF3 transgenic plants, respectively. Photographs were taken 12 d (right and middle of the lower panel) and 16 d (left of the lower panel) after inoculation. Arrows indicate TMV-inoculated leaves or mock-inoculated leaves. (This figure is available in colour at JXB online.)
Mentions: To determine whether overexpression of GmERF3 enhanced resistance to pathogens in transgenic tobacco plants, the bacterial pathogen Ralstonia solanacearum was injected into leaves of wild-type and transgenic plants, and symptom development was subsequently monitored for 7 d. As shown in Fig. 7A, all transgenic lines exhibited significantly reduced disease lesions and leaf bacterial numbers compared to wild-type plants. Approximately 50% inhibition of bacterial growth was detected in 35S::GmERF3 tobacco plants during the 7-d infection period (P <0.01). Line G2 showed stronger resistance than other transgenic lines tested.

Bottom Line: The GmERF3 protein fused to the GAL4 DNA-binding domain to activate transcription of reporter genes in yeast.Furthermore, overexpression of GmERF3 in transgenic tobacco led to higher levels of free proline and soluble carbohydrates compared to wild-type plants under drought conditions.The overall results suggested that GmERF3 as an AP2/ERF transcription factor may play dual roles in response to biotic and abiotic stresses in plants.

View Article: PubMed Central - PubMed

Affiliation: The National Key Facility for Crop Genetic Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
A new member of the AP2/ERF transcription factor family, GmERF3, was isolated from soybean. Sequence analysis showed that GmERF3 contained an AP2/ERF domain of 58 amino acids and two putative nuclear localization signal (NLS) domains. It belonged to a group IV protein in the ERF (ethylene response factor) subfamily as typified by a conserved N-terminal motif [MCGGAI(I/L)]. Expression of GmERF3 was induced by treatments with high salinity, drought, abscisic acid (ABA), salicylic acid (SA), jasmonic acid (JA), ethylene (ET), and soybean mosaic virus (SMV), whereas there was no significant GmERF3 mRNA accumulation under cold stress treatment. GmERF3 could bind to the GCC box and DRE/CRT element, and was targeted to the nucleus when transiently expressed in onion epidermal cells. The GmERF3 protein fused to the GAL4 DNA-binding domain to activate transcription of reporter genes in yeast. Ectopic expression of the GmERF3 gene in transgenic tobacco plants induced the expression of some PR genes and enhanced resistance against infection by Ralstonia solanacearum, Alternaria alternata, and tobacco mosaic virus (TMV), and gave tolerance to high salinity and dehydration stresses. Furthermore, overexpression of GmERF3 in transgenic tobacco led to higher levels of free proline and soluble carbohydrates compared to wild-type plants under drought conditions. The overall results suggested that GmERF3 as an AP2/ERF transcription factor may play dual roles in response to biotic and abiotic stresses in plants.

Show MeSH
Related in: MedlinePlus