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Banana Ovate family protein MaOFP1 and MADS-box protein MuMADS1 antagonistically regulated banana fruit ripening.

Liu J, Zhang J, Hu W, Miao H, Zhang J, Jia C, Wang Z, Xu B, Jin Z - PLoS ONE (2015)

Bottom Line: Meanwhile, MuMADS1 and MaOFP1 expression was highly stimulated and greatly suppressed, respectively, by exogenous ethylene.In contrast, MaOFP1 expression was highly stimulated while MuMADS1 was greatly suppressed by the ethylene competitor 1-methylcyclopropene (1-MCP).These results indicate that MuMADS1 and MaOFP1 are antagonistically regulated by ethylene and might play important roles in postharvest banana fruit ripening.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China.

ABSTRACT
The ovate family protein named MaOFP1 was identified in banana (Musa acuminata L.AAA) fruit by a yeast two-hybrid (Y2H) method using the banana MADS-box gene MuMADS1 as bait and a 2 day postharvest (DPH) banana fruit cDNA library as prey. The interaction between MuMADS1 and MaOFP1 was further confirmed by Y2H and Bimolecular Fluorescence Complementation (BiFC) methods, which showed that the MuMADS1 K domain interacted with MaOFP1. Real-time quantitative PCR evaluation of MuMADS1 and MaOFP1 expression patterns in banana showed that they are highly expressed in 0 DPH fruit, but present in low levels in the stem, which suggests that simultaneous but different expression patterns exist for both MuMADS1 and MaOFP1 in different tissues and developing fruits. Meanwhile, MuMADS1 and MaOFP1 expression was highly stimulated and greatly suppressed, respectively, by exogenous ethylene. In contrast, MaOFP1 expression was highly stimulated while MuMADS1 was greatly suppressed by the ethylene competitor 1-methylcyclopropene (1-MCP). These results indicate that MuMADS1 and MaOFP1 are antagonistically regulated by ethylene and might play important roles in postharvest banana fruit ripening.

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Interaction assay of MuMADS1 and MaOFP1 by the BiFC method.The free YFP protein (A) and MuMADS1-YN with MaOFP1-YC fusion proteins (B) were transiently expressed in onion epidermis cells and visualized with a ZEISS fluorescence microscope 24 h after bombardment with a gene gun. a and d: yellow fluorescence in cells, b and e: DAPI image, c and f: bright-field image. Panels C and D: Onion epidermis cells following bombardment with a gene gun as negative controls. C: MuMADS1-YN protein. D: MaOFP1-YC protein. g and j: yellow fluorescence. h and k: DAPI image. i and l: bright field image.
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pone.0123870.g002: Interaction assay of MuMADS1 and MaOFP1 by the BiFC method.The free YFP protein (A) and MuMADS1-YN with MaOFP1-YC fusion proteins (B) were transiently expressed in onion epidermis cells and visualized with a ZEISS fluorescence microscope 24 h after bombardment with a gene gun. a and d: yellow fluorescence in cells, b and e: DAPI image, c and f: bright-field image. Panels C and D: Onion epidermis cells following bombardment with a gene gun as negative controls. C: MuMADS1-YN protein. D: MaOFP1-YC protein. g and j: yellow fluorescence. h and k: DAPI image. i and l: bright field image.

Mentions: To analyze the interaction of MuMADS1 and MaOFP1 in vivo, MuMADS1 and MaOFP1 were fused in frame to the N-terminal and C-terminal fragments of YFP (YN and YC), respectively, under the control of the cauliflower mosaic virus 35S promoter (CaMV 35S). The two recombinant constructs were introduced into onion (Allium cepa) epidermal cells by the particle delivery method. Free YFP protein was distributed throughout the whole cell (nucleus and cytoplasm) (Fig 2A), whereas MuMADS1-YN with MaOFP1-YC accumulated mainly in the nucleus (Fig 2B). As negative controls, MuMADS1-YN and MaOFP1-YC were separately bombarded into onion epidermal cell, after which no fluorescence was detected (Fig 2C and 2D).


Banana Ovate family protein MaOFP1 and MADS-box protein MuMADS1 antagonistically regulated banana fruit ripening.

Liu J, Zhang J, Hu W, Miao H, Zhang J, Jia C, Wang Z, Xu B, Jin Z - PLoS ONE (2015)

Interaction assay of MuMADS1 and MaOFP1 by the BiFC method.The free YFP protein (A) and MuMADS1-YN with MaOFP1-YC fusion proteins (B) were transiently expressed in onion epidermis cells and visualized with a ZEISS fluorescence microscope 24 h after bombardment with a gene gun. a and d: yellow fluorescence in cells, b and e: DAPI image, c and f: bright-field image. Panels C and D: Onion epidermis cells following bombardment with a gene gun as negative controls. C: MuMADS1-YN protein. D: MaOFP1-YC protein. g and j: yellow fluorescence. h and k: DAPI image. i and l: bright field image.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4401719&req=5

pone.0123870.g002: Interaction assay of MuMADS1 and MaOFP1 by the BiFC method.The free YFP protein (A) and MuMADS1-YN with MaOFP1-YC fusion proteins (B) were transiently expressed in onion epidermis cells and visualized with a ZEISS fluorescence microscope 24 h after bombardment with a gene gun. a and d: yellow fluorescence in cells, b and e: DAPI image, c and f: bright-field image. Panels C and D: Onion epidermis cells following bombardment with a gene gun as negative controls. C: MuMADS1-YN protein. D: MaOFP1-YC protein. g and j: yellow fluorescence. h and k: DAPI image. i and l: bright field image.
Mentions: To analyze the interaction of MuMADS1 and MaOFP1 in vivo, MuMADS1 and MaOFP1 were fused in frame to the N-terminal and C-terminal fragments of YFP (YN and YC), respectively, under the control of the cauliflower mosaic virus 35S promoter (CaMV 35S). The two recombinant constructs were introduced into onion (Allium cepa) epidermal cells by the particle delivery method. Free YFP protein was distributed throughout the whole cell (nucleus and cytoplasm) (Fig 2A), whereas MuMADS1-YN with MaOFP1-YC accumulated mainly in the nucleus (Fig 2B). As negative controls, MuMADS1-YN and MaOFP1-YC were separately bombarded into onion epidermal cell, after which no fluorescence was detected (Fig 2C and 2D).

Bottom Line: Meanwhile, MuMADS1 and MaOFP1 expression was highly stimulated and greatly suppressed, respectively, by exogenous ethylene.In contrast, MaOFP1 expression was highly stimulated while MuMADS1 was greatly suppressed by the ethylene competitor 1-methylcyclopropene (1-MCP).These results indicate that MuMADS1 and MaOFP1 are antagonistically regulated by ethylene and might play important roles in postharvest banana fruit ripening.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China.

ABSTRACT
The ovate family protein named MaOFP1 was identified in banana (Musa acuminata L.AAA) fruit by a yeast two-hybrid (Y2H) method using the banana MADS-box gene MuMADS1 as bait and a 2 day postharvest (DPH) banana fruit cDNA library as prey. The interaction between MuMADS1 and MaOFP1 was further confirmed by Y2H and Bimolecular Fluorescence Complementation (BiFC) methods, which showed that the MuMADS1 K domain interacted with MaOFP1. Real-time quantitative PCR evaluation of MuMADS1 and MaOFP1 expression patterns in banana showed that they are highly expressed in 0 DPH fruit, but present in low levels in the stem, which suggests that simultaneous but different expression patterns exist for both MuMADS1 and MaOFP1 in different tissues and developing fruits. Meanwhile, MuMADS1 and MaOFP1 expression was highly stimulated and greatly suppressed, respectively, by exogenous ethylene. In contrast, MaOFP1 expression was highly stimulated while MuMADS1 was greatly suppressed by the ethylene competitor 1-methylcyclopropene (1-MCP). These results indicate that MuMADS1 and MaOFP1 are antagonistically regulated by ethylene and might play important roles in postharvest banana fruit ripening.

Show MeSH