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Functional Analysis of the Maize C-Repeat/DRE Motif-Binding Transcription Factor CBF3 Promoter in Response to Abiotic Stress.

Liu J, Wang F, Yu G, Zhang X, Jia C, Qin J, Pan H - Int J Mol Sci (2015)

Bottom Line: PZmCBF3 was activated by cold stress.The MYCCONSENSUSAT elements (CANNTG) were responsible for the ability of PZmCBF3 to respond to cold stress.The results of the present study suggest that PZmCBF3 might play a role in cold tolerance in maize.

View Article: PubMed Central - PubMed

Affiliation: College of Plant Sciences, Jilin University, Changchun 130062, China. jlliu@jlu.edu.cn.

ABSTRACT
The ZmCBF3 gene is a member of AP2/ERF transcription factor family, which is a large family of plant-specific transcription factors that share a well-conserved DNA-binding domain. To understand the regulatory mechanism of ZmCBF3 gene expression, we isolated and characterized the ZmCBF3 promoter (PZmCBF3). Three deletion fragments of PZmCBF3 were generated, C1-C3, from the translation start codon at position -1079, -638, and -234, and fused to the GUS reporter gene. Each deletion construct was analyzed by Agrobacterium-mediated stable transformation and expression in Arabidopsis thaliana. GUS expression assays indicated that the PZmCBF3 exhibited root-specific expression activity. A 234-bp fragment upstream of the ZmCBF3 gene conferred a high level of GUS activity in Arabidopsis. Some cis-acting elements involved in the down-regulation of gene expression were detected in the promoter, encompassing positions -1079 to -234. PZmCBF3 was activated by cold stress. The MYCCONSENSUSAT elements (CANNTG) were responsible for the ability of PZmCBF3 to respond to cold stress. The results of the present study suggest that PZmCBF3 might play a role in cold tolerance in maize.

No MeSH data available.


Related in: MedlinePlus

Tissue-specific expression analysis of GUS activity among different transgenic Arabidopsis lines. GUS activity was examined in six-week-old Arabidopsis T3 plants and wild type (WT). GUS activity was analyzed fluorometrically and expressed as nmoles 4-methylumbelliferone (MU)/mg protein/min. Data are expressed as the mean ± standard deviation of three independent assays involving Arabidopsis leaf extracts. The same letters indicate no significant differences at p ≤ 0.05 by Duncan’s test. Different letters (a–j) mean statistical differences between treatments.
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ijms-16-12131-f006: Tissue-specific expression analysis of GUS activity among different transgenic Arabidopsis lines. GUS activity was examined in six-week-old Arabidopsis T3 plants and wild type (WT). GUS activity was analyzed fluorometrically and expressed as nmoles 4-methylumbelliferone (MU)/mg protein/min. Data are expressed as the mean ± standard deviation of three independent assays involving Arabidopsis leaf extracts. The same letters indicate no significant differences at p ≤ 0.05 by Duncan’s test. Different letters (a–j) mean statistical differences between treatments.

Mentions: In our early experiments, histochemical staining was employed to determine GUS activity of the deletion promoters in transgenic Arabidopsis (Figure 4). In Figure 4, GUS staining was mainly detected in the stems and roots. The promoter apparently shows tissue-specific expression. To test this hypothesis, GUS activity of PZmCBF3 was determined in the transgenic Arabidopsis roots, stems, and leaves by fluorometric analyses. GUS activity of the C1 and C2 promoters was highest in the roots, followed by the stems (Figure 6). C3-mediated GUS activity was highest in the stems, which was 1.67-fold that of the root (Figure 6). These results were consistent with that of GUS staining.


Functional Analysis of the Maize C-Repeat/DRE Motif-Binding Transcription Factor CBF3 Promoter in Response to Abiotic Stress.

Liu J, Wang F, Yu G, Zhang X, Jia C, Qin J, Pan H - Int J Mol Sci (2015)

Tissue-specific expression analysis of GUS activity among different transgenic Arabidopsis lines. GUS activity was examined in six-week-old Arabidopsis T3 plants and wild type (WT). GUS activity was analyzed fluorometrically and expressed as nmoles 4-methylumbelliferone (MU)/mg protein/min. Data are expressed as the mean ± standard deviation of three independent assays involving Arabidopsis leaf extracts. The same letters indicate no significant differences at p ≤ 0.05 by Duncan’s test. Different letters (a–j) mean statistical differences between treatments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490434&req=5

ijms-16-12131-f006: Tissue-specific expression analysis of GUS activity among different transgenic Arabidopsis lines. GUS activity was examined in six-week-old Arabidopsis T3 plants and wild type (WT). GUS activity was analyzed fluorometrically and expressed as nmoles 4-methylumbelliferone (MU)/mg protein/min. Data are expressed as the mean ± standard deviation of three independent assays involving Arabidopsis leaf extracts. The same letters indicate no significant differences at p ≤ 0.05 by Duncan’s test. Different letters (a–j) mean statistical differences between treatments.
Mentions: In our early experiments, histochemical staining was employed to determine GUS activity of the deletion promoters in transgenic Arabidopsis (Figure 4). In Figure 4, GUS staining was mainly detected in the stems and roots. The promoter apparently shows tissue-specific expression. To test this hypothesis, GUS activity of PZmCBF3 was determined in the transgenic Arabidopsis roots, stems, and leaves by fluorometric analyses. GUS activity of the C1 and C2 promoters was highest in the roots, followed by the stems (Figure 6). C3-mediated GUS activity was highest in the stems, which was 1.67-fold that of the root (Figure 6). These results were consistent with that of GUS staining.

Bottom Line: PZmCBF3 was activated by cold stress.The MYCCONSENSUSAT elements (CANNTG) were responsible for the ability of PZmCBF3 to respond to cold stress.The results of the present study suggest that PZmCBF3 might play a role in cold tolerance in maize.

View Article: PubMed Central - PubMed

Affiliation: College of Plant Sciences, Jilin University, Changchun 130062, China. jlliu@jlu.edu.cn.

ABSTRACT
The ZmCBF3 gene is a member of AP2/ERF transcription factor family, which is a large family of plant-specific transcription factors that share a well-conserved DNA-binding domain. To understand the regulatory mechanism of ZmCBF3 gene expression, we isolated and characterized the ZmCBF3 promoter (PZmCBF3). Three deletion fragments of PZmCBF3 were generated, C1-C3, from the translation start codon at position -1079, -638, and -234, and fused to the GUS reporter gene. Each deletion construct was analyzed by Agrobacterium-mediated stable transformation and expression in Arabidopsis thaliana. GUS expression assays indicated that the PZmCBF3 exhibited root-specific expression activity. A 234-bp fragment upstream of the ZmCBF3 gene conferred a high level of GUS activity in Arabidopsis. Some cis-acting elements involved in the down-regulation of gene expression were detected in the promoter, encompassing positions -1079 to -234. PZmCBF3 was activated by cold stress. The MYCCONSENSUSAT elements (CANNTG) were responsible for the ability of PZmCBF3 to respond to cold stress. The results of the present study suggest that PZmCBF3 might play a role in cold tolerance in maize.

No MeSH data available.


Related in: MedlinePlus