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Cloning and Characterization of 5' Flanking Regulatory Sequences of AhLEC1B Gene from Arachis Hypogaea L.

Tang G, Xu P, Liu W, Liu Z, Shan L - PLoS ONE (2015)

Bottom Line: We further characterized the GUS expression patterns in the transgenic Arabidopsis lines.Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity.Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

View Article: PubMed Central - PubMed

Affiliation: Bio-Tech Research Centre, Shandong Academy of Agricultural Sciences / Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan, 250100, China.

ABSTRACT
LEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5' flanking regulatory sequence of AhLEC1B gene, a homolog of Arabidopsis LEC1, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5' terminal and/or 3' terminal deletion. We further characterized the GUS expression patterns in the transgenic Arabidopsis lines. Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity. Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

No MeSH data available.


Phylogenetic tree for peanut AhLEC1A and AhLEC1B, and the Arabidopsis NF-YB family.
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pone.0139213.g001: Phylogenetic tree for peanut AhLEC1A and AhLEC1B, and the Arabidopsis NF-YB family.

Mentions: In the Arabidopsis genome 13 NF-YB genes with distinctive expression patterns were found [3–5]. To predict the evolutionary relationship of AhLEC1A and AhLEC1B, a sequence comparison of AhLEC1A, AhLEC1B, and Arabidopsis NF-YB transcription factors was performed using MAGE 4.0. AhLEC1A, AhLEC1B, Arabidopsis NF-YB6 (L1L) and NF-YB9 (LEC1) have higher sequence similarity and group together (Fig 1). AhLEC1A and AhLEC1B share 95% sequence identity and diverge at only 12 amino acid sites. However, the expression profile of AhLEC1A was substantially different from that of AhLEC1B. AhLEC1A is expressed specifically in seeds during different developmental stages while AhLEC1B mRNA accumulates at higher levels in seeds as compared with roots, stems, rosettes, and flowers [14].


Cloning and Characterization of 5' Flanking Regulatory Sequences of AhLEC1B Gene from Arachis Hypogaea L.

Tang G, Xu P, Liu W, Liu Z, Shan L - PLoS ONE (2015)

Phylogenetic tree for peanut AhLEC1A and AhLEC1B, and the Arabidopsis NF-YB family.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591277&req=5

pone.0139213.g001: Phylogenetic tree for peanut AhLEC1A and AhLEC1B, and the Arabidopsis NF-YB family.
Mentions: In the Arabidopsis genome 13 NF-YB genes with distinctive expression patterns were found [3–5]. To predict the evolutionary relationship of AhLEC1A and AhLEC1B, a sequence comparison of AhLEC1A, AhLEC1B, and Arabidopsis NF-YB transcription factors was performed using MAGE 4.0. AhLEC1A, AhLEC1B, Arabidopsis NF-YB6 (L1L) and NF-YB9 (LEC1) have higher sequence similarity and group together (Fig 1). AhLEC1A and AhLEC1B share 95% sequence identity and diverge at only 12 amino acid sites. However, the expression profile of AhLEC1A was substantially different from that of AhLEC1B. AhLEC1A is expressed specifically in seeds during different developmental stages while AhLEC1B mRNA accumulates at higher levels in seeds as compared with roots, stems, rosettes, and flowers [14].

Bottom Line: We further characterized the GUS expression patterns in the transgenic Arabidopsis lines.Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity.Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

View Article: PubMed Central - PubMed

Affiliation: Bio-Tech Research Centre, Shandong Academy of Agricultural Sciences / Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan, 250100, China.

ABSTRACT
LEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5' flanking regulatory sequence of AhLEC1B gene, a homolog of Arabidopsis LEC1, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5' terminal and/or 3' terminal deletion. We further characterized the GUS expression patterns in the transgenic Arabidopsis lines. Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity. Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

No MeSH data available.