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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

Abiotic stress-induced GUS activity analysis of transgenic Arabidopsis lines. Six-week-old Arabidopsis T3 plants and wild type (WT) were subjected to ABA, high temperature (HT), cold (4 °C), and drought (20% PEG6000) treatments for 6 h. Control (CK) plants were treated with water. 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–g) mean statistical differences between treatments.
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ijms-16-12131-f005: Abiotic stress-induced GUS activity analysis of transgenic Arabidopsis lines. Six-week-old Arabidopsis T3 plants and wild type (WT) were subjected to ABA, high temperature (HT), cold (4 °C), and drought (20% PEG6000) treatments for 6 h. Control (CK) plants were treated with water. 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–g) mean statistical differences between treatments.

Mentions: Compared to the control leaves (untreated plants), GUS activity in C2-transformed leaves decreased by 0.547-fold after HT treatment (Figure 5). However, marginal induction of GUS activity was observed in C1- and C3-transformed Arabidopsis leaves. Under cold stress, C1- and C3-mediated GUS activity increased by 2.8- and 1.19-fold, respectively, whereas GUS activity of C2-transformed plants did not significantly change (Figure 5). As shown in Figure 5, GUS activity of deletion promoters slightly increased after ABA and PEG treatment. GUS activity of the C1 and C2 promoters increased by 1.15- and 1.21-fold respectively, after ABA treatment. C3-mediated GUS activity significantly increased (by about 1.22-fold) with PEG treatment.


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)

Abiotic stress-induced GUS activity analysis of transgenic Arabidopsis lines. Six-week-old Arabidopsis T3 plants and wild type (WT) were subjected to ABA, high temperature (HT), cold (4 °C), and drought (20% PEG6000) treatments for 6 h. Control (CK) plants were treated with water. 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–g) mean statistical differences between treatments.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-12131-f005: Abiotic stress-induced GUS activity analysis of transgenic Arabidopsis lines. Six-week-old Arabidopsis T3 plants and wild type (WT) were subjected to ABA, high temperature (HT), cold (4 °C), and drought (20% PEG6000) treatments for 6 h. Control (CK) plants were treated with water. 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–g) mean statistical differences between treatments.
Mentions: Compared to the control leaves (untreated plants), GUS activity in C2-transformed leaves decreased by 0.547-fold after HT treatment (Figure 5). However, marginal induction of GUS activity was observed in C1- and C3-transformed Arabidopsis leaves. Under cold stress, C1- and C3-mediated GUS activity increased by 2.8- and 1.19-fold, respectively, whereas GUS activity of C2-transformed plants did not significantly change (Figure 5). As shown in Figure 5, GUS activity of deletion promoters slightly increased after ABA and PEG treatment. GUS activity of the C1 and C2 promoters increased by 1.15- and 1.21-fold respectively, after ABA treatment. C3-mediated GUS activity significantly increased (by about 1.22-fold) with PEG treatment.

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