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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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Y2H assays.(A) A schematic diagram illustrating the MuMADS1 cDNA fragments encoding different portions of M, I, K and C that were fused to DNA sequences encoding the GAL4 DNA binding domain in the yeast vector pGBKT7. MaOFP1 was cloned into pGADT7 as prey. (B) Positive interactions were determined through auxotrophic selection media SD/-Ade/-His-Leu/-Trp and SD/-Ade/-His-Leu/-Trp+ x-α-gal. The pGBKT7-53 vector used as a positive control interacted with the pGADT7-T-antigen while pGBKT7 empty vector as a negative control interacted with the pGADT7 empty vector.
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pone.0123870.g003: Y2H assays.(A) A schematic diagram illustrating the MuMADS1 cDNA fragments encoding different portions of M, I, K and C that were fused to DNA sequences encoding the GAL4 DNA binding domain in the yeast vector pGBKT7. MaOFP1 was cloned into pGADT7 as prey. (B) Positive interactions were determined through auxotrophic selection media SD/-Ade/-His-Leu/-Trp and SD/-Ade/-His-Leu/-Trp+ x-α-gal. The pGBKT7-53 vector used as a positive control interacted with the pGADT7-T-antigen while pGBKT7 empty vector as a negative control interacted with the pGADT7 empty vector.

Mentions: To understand which domain of MuMADS1 interacted with MaOFP1, MuMADS1 was divided into four domains of M, I, K and C according to the sequence analysis. These four domains were each cloned into the pGBKT7 vector to fuse with the BD (bait domain). When the bait vectors containing the M, I, K and C domains and the prey vector containing MaOFP1 were transformed into yeast strain AH109 and grown on auxotrophic selection media SD/-Ade/-His-Leu/-Trp+x-α-gal, blue clones appeared only for K+MaOFP1 and the positive control (pGBKT7-53+ pGADT7-T-antigen) as compared to the negative control. Other interactions of M + MaOFP1, I + MaOFP1 and C+ MaOFP1 yielded no blue clone growth. This result indicated that the K domain of MuMADS1 interacts with MaOFP1 in yeast (Fig 3A and 3B).


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)

Y2H assays.(A) A schematic diagram illustrating the MuMADS1 cDNA fragments encoding different portions of M, I, K and C that were fused to DNA sequences encoding the GAL4 DNA binding domain in the yeast vector pGBKT7. MaOFP1 was cloned into pGADT7 as prey. (B) Positive interactions were determined through auxotrophic selection media SD/-Ade/-His-Leu/-Trp and SD/-Ade/-His-Leu/-Trp+ x-α-gal. The pGBKT7-53 vector used as a positive control interacted with the pGADT7-T-antigen while pGBKT7 empty vector as a negative control interacted with the pGADT7 empty vector.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401719&req=5

pone.0123870.g003: Y2H assays.(A) A schematic diagram illustrating the MuMADS1 cDNA fragments encoding different portions of M, I, K and C that were fused to DNA sequences encoding the GAL4 DNA binding domain in the yeast vector pGBKT7. MaOFP1 was cloned into pGADT7 as prey. (B) Positive interactions were determined through auxotrophic selection media SD/-Ade/-His-Leu/-Trp and SD/-Ade/-His-Leu/-Trp+ x-α-gal. The pGBKT7-53 vector used as a positive control interacted with the pGADT7-T-antigen while pGBKT7 empty vector as a negative control interacted with the pGADT7 empty vector.
Mentions: To understand which domain of MuMADS1 interacted with MaOFP1, MuMADS1 was divided into four domains of M, I, K and C according to the sequence analysis. These four domains were each cloned into the pGBKT7 vector to fuse with the BD (bait domain). When the bait vectors containing the M, I, K and C domains and the prey vector containing MaOFP1 were transformed into yeast strain AH109 and grown on auxotrophic selection media SD/-Ade/-His-Leu/-Trp+x-α-gal, blue clones appeared only for K+MaOFP1 and the positive control (pGBKT7-53+ pGADT7-T-antigen) as compared to the negative control. Other interactions of M + MaOFP1, I + MaOFP1 and C+ MaOFP1 yielded no blue clone growth. This result indicated that the K domain of MuMADS1 interacts with MaOFP1 in yeast (Fig 3A and 3B).

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