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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 TaGAAP promoter in wheat seedlings by promoter truncation and site-directed mutagenesis of TaGAAPE1. A xylanase (XYNA) was used as a reporter for quantitative measurement of expression levels of reporter constructs driven by TaGAAP and its mutant promoters with or without the TaHsfA6f effector construct (Ubi1A6f). Values are means±SD of 3–4 biological replicates. Ubi1:GUS+ was used as a control gene for normalisation of transformation efficiency. Relative expression levels of the reporter gene are expressed as the ratio of XYNA to GUS activity relative to that of GAAPxynA reporter construct with the TaHsfA6f effector construct, which is arbitrarily set as 1. GAAP–763xynA contains a TaGAAP promoter fragment of 763bp upstream of the translation initiation codon. GAAP–763ΔHSExynA contains a mutated TaGAAPE1.
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Figure 7: Functional analysis of the TaGAAP promoter in wheat seedlings by promoter truncation and site-directed mutagenesis of TaGAAPE1. A xylanase (XYNA) was used as a reporter for quantitative measurement of expression levels of reporter constructs driven by TaGAAP and its mutant promoters with or without the TaHsfA6f effector construct (Ubi1A6f). Values are means±SD of 3–4 biological replicates. Ubi1:GUS+ was used as a control gene for normalisation of transformation efficiency. Relative expression levels of the reporter gene are expressed as the ratio of XYNA to GUS activity relative to that of GAAPxynA reporter construct with the TaHsfA6f effector construct, which is arbitrarily set as 1. GAAP–763xynA contains a TaGAAP promoter fragment of 763bp upstream of the translation initiation codon. GAAP–763ΔHSExynA contains a mutated TaGAAPE1.

Mentions: The promoter truncation and HSE mutation were also performed for the TaGAAP promoter using a xylanase (xynA) gene as a reporter for quantitative analysis of TaHsfA6f transactivation activity (Fig. 7). TaGAAPE1 is located downstream of a putative TATA box in the TaGAAP promoter. Removal of a 282-bp upstream fragment (containing TaGAAPE2) in the 1045-bp TaGAAP promoter (GAAP–763xynA construct) did not significantly affect its promoter activity induced by the Ubi1 promoter-driven TaHsfA6f effector construct (Fig. 7). Mutation of the TaGAAPE1 element (GAAGCCTCCTGAATCTTC) by site-direct mutagenesis to GAAGCCTCCTGGTACCAC resulted in a GAAP–763ΔHSExynA reporter construct that was no longer transactivated by TaHsfA6f (Fig. 7). These data indicate that the TaGAAPE1 element is a functional HSE required for TaGAAP transactivation by TaHsfA6f.


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 TaGAAP promoter in wheat seedlings by promoter truncation and site-directed mutagenesis of TaGAAPE1. A xylanase (XYNA) was used as a reporter for quantitative measurement of expression levels of reporter constructs driven by TaGAAP and its mutant promoters with or without the TaHsfA6f effector construct (Ubi1A6f). Values are means±SD of 3–4 biological replicates. Ubi1:GUS+ was used as a control gene for normalisation of transformation efficiency. Relative expression levels of the reporter gene are expressed as the ratio of XYNA to GUS activity relative to that of GAAPxynA reporter construct with the TaHsfA6f effector construct, which is arbitrarily set as 1. GAAP–763xynA contains a TaGAAP promoter fragment of 763bp upstream of the translation initiation codon. GAAP–763ΔHSExynA contains a mutated TaGAAPE1.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4321556&req=5

Figure 7: Functional analysis of the TaGAAP promoter in wheat seedlings by promoter truncation and site-directed mutagenesis of TaGAAPE1. A xylanase (XYNA) was used as a reporter for quantitative measurement of expression levels of reporter constructs driven by TaGAAP and its mutant promoters with or without the TaHsfA6f effector construct (Ubi1A6f). Values are means±SD of 3–4 biological replicates. Ubi1:GUS+ was used as a control gene for normalisation of transformation efficiency. Relative expression levels of the reporter gene are expressed as the ratio of XYNA to GUS activity relative to that of GAAPxynA reporter construct with the TaHsfA6f effector construct, which is arbitrarily set as 1. GAAP–763xynA contains a TaGAAP promoter fragment of 763bp upstream of the translation initiation codon. GAAP–763ΔHSExynA contains a mutated TaGAAPE1.
Mentions: The promoter truncation and HSE mutation were also performed for the TaGAAP promoter using a xylanase (xynA) gene as a reporter for quantitative analysis of TaHsfA6f transactivation activity (Fig. 7). TaGAAPE1 is located downstream of a putative TATA box in the TaGAAP promoter. Removal of a 282-bp upstream fragment (containing TaGAAPE2) in the 1045-bp TaGAAP promoter (GAAP–763xynA construct) did not significantly affect its promoter activity induced by the Ubi1 promoter-driven TaHsfA6f effector construct (Fig. 7). Mutation of the TaGAAPE1 element (GAAGCCTCCTGAATCTTC) by site-direct mutagenesis to GAAGCCTCCTGGTACCAC resulted in a GAAP–763ΔHSExynA reporter construct that was no longer transactivated by TaHsfA6f (Fig. 7). These data indicate that the TaGAAPE1 element is a functional HSE required for TaGAAP transactivation by TaHsfA6f.

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