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Histone acetylation accompanied with promoter sequences displaying differential expression profiles of B-class MADS-box genes for phalaenopsis floral morphogenesis.

Hsu CC, Wu PS, Chen TC, Yu CW, Tsai WC, Wu K, Wu WL, Chen WH, Chen HH - PLoS ONE (2014)

Bottom Line: The amplified promoter sequences of PeMADS2∼6 could drive beta-glucuronidase (GUS) gene expression in all floral organs, similar to their expression at the floral primordia stage.The promoter sequence of PeMADS4, exclusively expressed in lip and column, showed a 1.6∼3-fold higher expression in lip/column than in sepal/petal.Furthermore, we noted a 4.9-fold increase in histone acetylation (H3K9K14ac) in the translation start region of PeMADS4 in lip as compared in petal.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences, National Cheng Kung University, Tainan, Taiwan.

ABSTRACT
Five B-class MADS-box genes, including four APETALA3 (AP3)-like PeMADS2∼5 and one PISTILLATA (PI)-like PeMADS6, specify the spectacular flower morphology in orchids. The PI-like PeMADS6 ubiquitously expresses in all floral organs. The four AP3-like genes, resulted from two duplication events, express ubiquitously at floral primordia and early floral organ stages, but show distinct expression profiles at late floral organ primordia and floral bud stages. Here, we isolated the upstream sequences of PeMADS2∼6 and studied the regulatory mechanism for their distinct gene expression. Phylogenetic footprinting analysis of the 1.3-kb upstream sequences of AP3-like PeMADS2∼5 showed that their promoter regions have sufficiently diverged and contributed to their subfunctionalization. The amplified promoter sequences of PeMADS2∼6 could drive beta-glucuronidase (GUS) gene expression in all floral organs, similar to their expression at the floral primordia stage. The promoter sequence of PeMADS4, exclusively expressed in lip and column, showed a 1.6∼3-fold higher expression in lip/column than in sepal/petal. Furthermore, we noted a 4.9-fold increase in histone acetylation (H3K9K14ac) in the translation start region of PeMADS4 in lip as compared in petal. All these results suggest that the regulation via the upstream sequences and increased H3K9K14ac level may act synergistically to display distinct expression profiles of the AP3-like genes at late floral organ primordia stage for Phalaenopsis floral morphogenesis.

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Functional analysis of serial deletions of PeMADS4 promoter.(A) Serial deletion constructs of PeMADS4 promoter. (B–Q) Histochemical assay of flower organs bombarded with serial deletions of PeMADS4 promoter shown in the order of pBI-Pe4p-2313 (B–E), pBI-Pe4p-1497 (F–I), pBI-Pe4p-935 (J–M) and pBI-Pe4p-375 (N–Q). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  = 0.5 mm. (R) Dual luciferase assay of the serial deletions of PeMADS4 promoter. The same letters above the bars are not statistically different by T-test analysis (p<0.01). Data are mean ± SD (n = 6). All constructs were analyzed for promoter activities of driving luciferase expression by bombardment into two floral buds, and results are representative of three independent bombardment experiments.
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pone-0106033-g005: Functional analysis of serial deletions of PeMADS4 promoter.(A) Serial deletion constructs of PeMADS4 promoter. (B–Q) Histochemical assay of flower organs bombarded with serial deletions of PeMADS4 promoter shown in the order of pBI-Pe4p-2313 (B–E), pBI-Pe4p-1497 (F–I), pBI-Pe4p-935 (J–M) and pBI-Pe4p-375 (N–Q). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  = 0.5 mm. (R) Dual luciferase assay of the serial deletions of PeMADS4 promoter. The same letters above the bars are not statistically different by T-test analysis (p<0.01). Data are mean ± SD (n = 6). All constructs were analyzed for promoter activities of driving luciferase expression by bombardment into two floral buds, and results are representative of three independent bombardment experiments.

Mentions: The four AP3-like PeMADS2∼5 genes express differentially in the floral organs with distinct patterns [17]. Two to five serial deletion clones for the upstream regulatory sequences of PeMADS2∼5 were constructed for GUS expression assay. Similarly, most serial deletion constructs could drive GUS expression in all four floral organs (Fig. 5B–Q, S1–S3 Figures), resembled to the gene expression patterns at the early floral organ primordia stage. Among them, the minimal promoters of PeMADS2∼5 were found to be 291 bp, 407 bp, 375 bp, and 122 bp of their upstream regulatory sequences, respectively (Fig. 5N–Q, S1–S3 Figures).


Histone acetylation accompanied with promoter sequences displaying differential expression profiles of B-class MADS-box genes for phalaenopsis floral morphogenesis.

Hsu CC, Wu PS, Chen TC, Yu CW, Tsai WC, Wu K, Wu WL, Chen WH, Chen HH - PLoS ONE (2014)

Functional analysis of serial deletions of PeMADS4 promoter.(A) Serial deletion constructs of PeMADS4 promoter. (B–Q) Histochemical assay of flower organs bombarded with serial deletions of PeMADS4 promoter shown in the order of pBI-Pe4p-2313 (B–E), pBI-Pe4p-1497 (F–I), pBI-Pe4p-935 (J–M) and pBI-Pe4p-375 (N–Q). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  = 0.5 mm. (R) Dual luciferase assay of the serial deletions of PeMADS4 promoter. The same letters above the bars are not statistically different by T-test analysis (p<0.01). Data are mean ± SD (n = 6). All constructs were analyzed for promoter activities of driving luciferase expression by bombardment into two floral buds, and results are representative of three independent bombardment experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263434&req=5

pone-0106033-g005: Functional analysis of serial deletions of PeMADS4 promoter.(A) Serial deletion constructs of PeMADS4 promoter. (B–Q) Histochemical assay of flower organs bombarded with serial deletions of PeMADS4 promoter shown in the order of pBI-Pe4p-2313 (B–E), pBI-Pe4p-1497 (F–I), pBI-Pe4p-935 (J–M) and pBI-Pe4p-375 (N–Q). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  = 0.5 mm. (R) Dual luciferase assay of the serial deletions of PeMADS4 promoter. The same letters above the bars are not statistically different by T-test analysis (p<0.01). Data are mean ± SD (n = 6). All constructs were analyzed for promoter activities of driving luciferase expression by bombardment into two floral buds, and results are representative of three independent bombardment experiments.
Mentions: The four AP3-like PeMADS2∼5 genes express differentially in the floral organs with distinct patterns [17]. Two to five serial deletion clones for the upstream regulatory sequences of PeMADS2∼5 were constructed for GUS expression assay. Similarly, most serial deletion constructs could drive GUS expression in all four floral organs (Fig. 5B–Q, S1–S3 Figures), resembled to the gene expression patterns at the early floral organ primordia stage. Among them, the minimal promoters of PeMADS2∼5 were found to be 291 bp, 407 bp, 375 bp, and 122 bp of their upstream regulatory sequences, respectively (Fig. 5N–Q, S1–S3 Figures).

Bottom Line: The amplified promoter sequences of PeMADS2∼6 could drive beta-glucuronidase (GUS) gene expression in all floral organs, similar to their expression at the floral primordia stage.The promoter sequence of PeMADS4, exclusively expressed in lip and column, showed a 1.6∼3-fold higher expression in lip/column than in sepal/petal.Furthermore, we noted a 4.9-fold increase in histone acetylation (H3K9K14ac) in the translation start region of PeMADS4 in lip as compared in petal.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences, National Cheng Kung University, Tainan, Taiwan.

ABSTRACT
Five B-class MADS-box genes, including four APETALA3 (AP3)-like PeMADS2∼5 and one PISTILLATA (PI)-like PeMADS6, specify the spectacular flower morphology in orchids. The PI-like PeMADS6 ubiquitously expresses in all floral organs. The four AP3-like genes, resulted from two duplication events, express ubiquitously at floral primordia and early floral organ stages, but show distinct expression profiles at late floral organ primordia and floral bud stages. Here, we isolated the upstream sequences of PeMADS2∼6 and studied the regulatory mechanism for their distinct gene expression. Phylogenetic footprinting analysis of the 1.3-kb upstream sequences of AP3-like PeMADS2∼5 showed that their promoter regions have sufficiently diverged and contributed to their subfunctionalization. The amplified promoter sequences of PeMADS2∼6 could drive beta-glucuronidase (GUS) gene expression in all floral organs, similar to their expression at the floral primordia stage. The promoter sequence of PeMADS4, exclusively expressed in lip and column, showed a 1.6∼3-fold higher expression in lip/column than in sepal/petal. Furthermore, we noted a 4.9-fold increase in histone acetylation (H3K9K14ac) in the translation start region of PeMADS4 in lip as compared in petal. All these results suggest that the regulation via the upstream sequences and increased H3K9K14ac level may act synergistically to display distinct expression profiles of the AP3-like genes at late floral organ primordia stage for Phalaenopsis floral morphogenesis.

Show MeSH
Related in: MedlinePlus