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TaHsfA6f is a transcriptional activator that regulates a suite of heat stress protection genes in wheat (Triticum aestivum L.) including previously unknown Hsf targets.

Xue GP, Drenth J, McIntyre CL - J. Exp. Bot. (2014)

Bottom Line: Transgenic wheat plants overexpressing TaHsfA6f showed improved thermotolerance.Promoter truncation and mutagenesis analyses identified TaHsfA6f-binding elements that were responsible for transactivation of TaHSP90.1-A1 and TaGAAP by TaHsfA6f.These data suggest that TaHsfA6f is a transcriptional activator that directly regulates TaHSP, TaGAAP, and TaRof1 genes in wheat and its gene regulatory network has a positive impact on thermotolerance.

View Article: PubMed Central - PubMed

Affiliation: CSIRO Plant Industry, 306 Carmody Road, St Lucia, Qld 4067, Australia gang-ping.xue@csiro.au.

No MeSH data available.


Related in: MedlinePlus

Functional analysis of the TaHSP90.1-A1 promoter and TaHSP90.1E1 HSE (GAAGCTTCGGGAA) in wheat seedlings. (A) Reporter gene constructs. A short TaHSP90.1-A1 promoter (sHSP90) has a 328-bp fragment upstream of the translational start codon, which contains TaHSP90.1E1. The sequence upstream of the TATA box of TaHSP90.1-A1 promoter is shown. The ΔHSE90 promoter starts immediately downstream of the TaHSP90.1E1 HSE. HSE90 construct contains three TaHSP90.1E1 HSE repeats, which are added immediately upstream of the ΔHSE90 promoter. The HSE90-miniDhn6 construct was made by adding three TaHSP90.1E1 HSE repeats to a minimal promoter of the drought-inducible Dhn6 gene. (B) Transactivation analysis. A GFP reporter gene was introduced into the shoots of wheat seedlings with or without the TaHsfA6f effect construct (Ubi1A6f). A Ubi1:GUS+ reporter gene was also co-introduced. Illustration of GUS foci is given only when GFP reporter expression is essentially undetectable in the tissue section.
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Figure 6: Functional analysis of the TaHSP90.1-A1 promoter and TaHSP90.1E1 HSE (GAAGCTTCGGGAA) in wheat seedlings. (A) Reporter gene constructs. A short TaHSP90.1-A1 promoter (sHSP90) has a 328-bp fragment upstream of the translational start codon, which contains TaHSP90.1E1. The sequence upstream of the TATA box of TaHSP90.1-A1 promoter is shown. The ΔHSE90 promoter starts immediately downstream of the TaHSP90.1E1 HSE. HSE90 construct contains three TaHSP90.1E1 HSE repeats, which are added immediately upstream of the ΔHSE90 promoter. The HSE90-miniDhn6 construct was made by adding three TaHSP90.1E1 HSE repeats to a minimal promoter of the drought-inducible Dhn6 gene. (B) Transactivation analysis. A GFP reporter gene was introduced into the shoots of wheat seedlings with or without the TaHsfA6f effect construct (Ubi1A6f). A Ubi1:GUS+ reporter gene was also co-introduced. Illustration of GUS foci is given only when GFP reporter expression is essentially undetectable in the tissue section.

Mentions: To assess whether transactivation of the reporter genes by TaHsfA6f occurs through its binding to HSE present in the promoters of the reporter genes, promoter truncation and mutation analyses of TaHSP90.1-A1 and TaGAAP were performed. A truncated TaHSP90.1-A1 promoter (a 328-bp fragment, named as sHSP90) containing a TaHSP90.1E1 HSE was still functional for TaHsfA6f-mediated transactivation of the reporter gene (Fig. 6). A further deletion of a 64-bp fragment with the removal of TaHSP90.1E1 (ΔHSE90) abolished the TaHsfA6f-mediated transactivation of the GFP reporter gene (Fig. 6B) or the heat-inducible promoter activity as shown previously (Xue et al., 2014). Addition of the TaHSP90.1E1 element to the ΔHSE90 promoter-driven GFP reporter gene restored transactivation by TaHsfA6f. TaHsfA6f was also able to transactivate a reporter gene driven by a minimal promoter (MiniDhn6) from a barley drought-inducible promoter (Dhn6) with the addition of TaHSP90.1E1 (HSE90-MiniDhn6) (Fig. 6).


TaHsfA6f is a transcriptional activator that regulates a suite of heat stress protection genes in wheat (Triticum aestivum L.) including previously unknown Hsf targets.

Xue GP, Drenth J, McIntyre CL - J. Exp. Bot. (2014)

Functional analysis of the TaHSP90.1-A1 promoter and TaHSP90.1E1 HSE (GAAGCTTCGGGAA) in wheat seedlings. (A) Reporter gene constructs. A short TaHSP90.1-A1 promoter (sHSP90) has a 328-bp fragment upstream of the translational start codon, which contains TaHSP90.1E1. The sequence upstream of the TATA box of TaHSP90.1-A1 promoter is shown. The ΔHSE90 promoter starts immediately downstream of the TaHSP90.1E1 HSE. HSE90 construct contains three TaHSP90.1E1 HSE repeats, which are added immediately upstream of the ΔHSE90 promoter. The HSE90-miniDhn6 construct was made by adding three TaHSP90.1E1 HSE repeats to a minimal promoter of the drought-inducible Dhn6 gene. (B) Transactivation analysis. A GFP reporter gene was introduced into the shoots of wheat seedlings with or without the TaHsfA6f effect construct (Ubi1A6f). A Ubi1:GUS+ reporter gene was also co-introduced. Illustration of GUS foci is given only when GFP reporter expression is essentially undetectable in the tissue section.
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Related In: Results  -  Collection

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Figure 6: Functional analysis of the TaHSP90.1-A1 promoter and TaHSP90.1E1 HSE (GAAGCTTCGGGAA) in wheat seedlings. (A) Reporter gene constructs. A short TaHSP90.1-A1 promoter (sHSP90) has a 328-bp fragment upstream of the translational start codon, which contains TaHSP90.1E1. The sequence upstream of the TATA box of TaHSP90.1-A1 promoter is shown. The ΔHSE90 promoter starts immediately downstream of the TaHSP90.1E1 HSE. HSE90 construct contains three TaHSP90.1E1 HSE repeats, which are added immediately upstream of the ΔHSE90 promoter. The HSE90-miniDhn6 construct was made by adding three TaHSP90.1E1 HSE repeats to a minimal promoter of the drought-inducible Dhn6 gene. (B) Transactivation analysis. A GFP reporter gene was introduced into the shoots of wheat seedlings with or without the TaHsfA6f effect construct (Ubi1A6f). A Ubi1:GUS+ reporter gene was also co-introduced. Illustration of GUS foci is given only when GFP reporter expression is essentially undetectable in the tissue section.
Mentions: To assess whether transactivation of the reporter genes by TaHsfA6f occurs through its binding to HSE present in the promoters of the reporter genes, promoter truncation and mutation analyses of TaHSP90.1-A1 and TaGAAP were performed. A truncated TaHSP90.1-A1 promoter (a 328-bp fragment, named as sHSP90) containing a TaHSP90.1E1 HSE was still functional for TaHsfA6f-mediated transactivation of the reporter gene (Fig. 6). A further deletion of a 64-bp fragment with the removal of TaHSP90.1E1 (ΔHSE90) abolished the TaHsfA6f-mediated transactivation of the GFP reporter gene (Fig. 6B) or the heat-inducible promoter activity as shown previously (Xue et al., 2014). Addition of the TaHSP90.1E1 element to the ΔHSE90 promoter-driven GFP reporter gene restored transactivation by TaHsfA6f. TaHsfA6f was also able to transactivate a reporter gene driven by a minimal promoter (MiniDhn6) from a barley drought-inducible promoter (Dhn6) with the addition of TaHSP90.1E1 (HSE90-MiniDhn6) (Fig. 6).

Bottom Line: Transgenic wheat plants overexpressing TaHsfA6f showed improved thermotolerance.Promoter truncation and mutagenesis analyses identified TaHsfA6f-binding elements that were responsible for transactivation of TaHSP90.1-A1 and TaGAAP by TaHsfA6f.These data suggest that TaHsfA6f is a transcriptional activator that directly regulates TaHSP, TaGAAP, and TaRof1 genes in wheat and its gene regulatory network has a positive impact on thermotolerance.

View Article: PubMed Central - PubMed

Affiliation: CSIRO Plant Industry, 306 Carmody Road, St Lucia, Qld 4067, Australia gang-ping.xue@csiro.au.

No MeSH data available.


Related in: MedlinePlus