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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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GUS histochemical staining for the promoter activities of PeMADS2∼6.Histochemical assay of GUS expression in floral organs shown in the order of pBI-Pe2p-3249 (A–D), pBI-Pe3p-1293 (E–H), pBI-Pe4p-3303 (I–L), pBI-Pe5p-2062 (M–P), pBI-Pe6p-1514 (Q–T), pBI221 (U–X) and pBI-PL (Y-AB). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  =  0.5 mm.
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pone-0106033-g003: GUS histochemical staining for the promoter activities of PeMADS2∼6.Histochemical assay of GUS expression in floral organs shown in the order of pBI-Pe2p-3249 (A–D), pBI-Pe3p-1293 (E–H), pBI-Pe4p-3303 (I–L), pBI-Pe5p-2062 (M–P), pBI-Pe6p-1514 (Q–T), pBI221 (U–X) and pBI-PL (Y-AB). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  =  0.5 mm.

Mentions: We examined the promoter activities of PeMADS2∼6 fragments with length of 3,249 bp, 1,293 bp, 3,303 bp, 2,062 bp, and 1,514 bp, respectively, for their ability to drive GUS expression by bombarding them into 1.5-cm floral buds of P. aphrodite subsp. formosana. Notably, all five PCR-amplified promoter fragments could drive GUS expression in the floral organs examined (Fig. 3A–T), similar to their expression patterns at the early floral primordia stage [20]. Moreover, serial deletion clones of the upstream sequences of PeMADS2∼6 were constructed for GUS expression assay.


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)

GUS histochemical staining for the promoter activities of PeMADS2∼6.Histochemical assay of GUS expression in floral organs shown in the order of pBI-Pe2p-3249 (A–D), pBI-Pe3p-1293 (E–H), pBI-Pe4p-3303 (I–L), pBI-Pe5p-2062 (M–P), pBI-Pe6p-1514 (Q–T), pBI221 (U–X) and pBI-PL (Y-AB). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  =  0.5 mm.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0106033-g003: GUS histochemical staining for the promoter activities of PeMADS2∼6.Histochemical assay of GUS expression in floral organs shown in the order of pBI-Pe2p-3249 (A–D), pBI-Pe3p-1293 (E–H), pBI-Pe4p-3303 (I–L), pBI-Pe5p-2062 (M–P), pBI-Pe6p-1514 (Q–T), pBI221 (U–X) and pBI-PL (Y-AB). Constructs were bombarded into four independent floral buds, and results are representative of three independent bombardment experiments. Scale bar  =  0.5 mm.
Mentions: We examined the promoter activities of PeMADS2∼6 fragments with length of 3,249 bp, 1,293 bp, 3,303 bp, 2,062 bp, and 1,514 bp, respectively, for their ability to drive GUS expression by bombarding them into 1.5-cm floral buds of P. aphrodite subsp. formosana. Notably, all five PCR-amplified promoter fragments could drive GUS expression in the floral organs examined (Fig. 3A–T), similar to their expression patterns at the early floral primordia stage [20]. Moreover, serial deletion clones of the upstream sequences of PeMADS2∼6 were constructed for GUS expression assay.

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