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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

Basal expression analysis of GUS activity in transgenic Arabidopsis lines. 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 of Arabidopsis leaf extracts. Six-week-old Arabidopsis T3 plants and wild type (WT) were used for this analysis. Same letters indicate no significant differences at p ≤ 0.05 by Duncan test. C1(−1,079), C2(−638), and C3(−234) represent different deletion promoters. Different letters (a–d) mean statistical differences between treatments.
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ijms-16-12131-f003: Basal expression analysis of GUS activity in transgenic Arabidopsis lines. 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 of Arabidopsis leaf extracts. Six-week-old Arabidopsis T3 plants and wild type (WT) were used for this analysis. Same letters indicate no significant differences at p ≤ 0.05 by Duncan test. C1(−1,079), C2(−638), and C3(−234) represent different deletion promoters. Different letters (a–d) mean statistical differences between treatments.

Mentions: To determine the minimal necessary region of PZmCBF3, three promoter deletion fragments were fused with the GUS reporter gene in pCAMBIA1301 for agro-infiltration into Arabidopsis (Figure 2). The expression of each PZmCBF3:GUS fusion gene was examined in the leaves of 15 independent T3 transgenic Arabidopsis plants. As shown in Figure 3, deletion constructs with C3 showed much higher GUS activity than that of the C1 and C2 promoters in the leaves of transgenic Arabidopsis plants (Figure 3 and Figure 4). C1 and C2 promoter-mediated GUS activity showed no significant difference. Compared to GUS activity driven by cauliflower mosaic virus (CaMV) 35S promoter (as positive control), PZmCBF3 showed lower activity in leaves. In addition, C3-mediated GUS activity was accounting for only 13% of the CaMV 35S promoter-mediated GUS activity. These results indicated that the leaves of the T3 transgenic Arabidopsis plants had very low PZmCBF3 activity.


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)

Basal expression analysis of GUS activity in transgenic Arabidopsis lines. 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 of Arabidopsis leaf extracts. Six-week-old Arabidopsis T3 plants and wild type (WT) were used for this analysis. Same letters indicate no significant differences at p ≤ 0.05 by Duncan test. C1(−1,079), C2(−638), and C3(−234) represent different deletion promoters. Different letters (a–d) mean statistical differences between treatments.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-12131-f003: Basal expression analysis of GUS activity in transgenic Arabidopsis lines. 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 of Arabidopsis leaf extracts. Six-week-old Arabidopsis T3 plants and wild type (WT) were used for this analysis. Same letters indicate no significant differences at p ≤ 0.05 by Duncan test. C1(−1,079), C2(−638), and C3(−234) represent different deletion promoters. Different letters (a–d) mean statistical differences between treatments.
Mentions: To determine the minimal necessary region of PZmCBF3, three promoter deletion fragments were fused with the GUS reporter gene in pCAMBIA1301 for agro-infiltration into Arabidopsis (Figure 2). The expression of each PZmCBF3:GUS fusion gene was examined in the leaves of 15 independent T3 transgenic Arabidopsis plants. As shown in Figure 3, deletion constructs with C3 showed much higher GUS activity than that of the C1 and C2 promoters in the leaves of transgenic Arabidopsis plants (Figure 3 and Figure 4). C1 and C2 promoter-mediated GUS activity showed no significant difference. Compared to GUS activity driven by cauliflower mosaic virus (CaMV) 35S promoter (as positive control), PZmCBF3 showed lower activity in leaves. In addition, C3-mediated GUS activity was accounting for only 13% of the CaMV 35S promoter-mediated GUS activity. These results indicated that the leaves of the T3 transgenic Arabidopsis plants had very low PZmCBF3 activity.

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