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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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Sequence analysis of MaOFP1.(A) Sequence alignments of MaOFP1 with four other OFPs from the banana A genome. MaAchr7, 5, 6 and 9 represent OFPs located on chromosomes 7, 5, 6 and 9, respectively. Identical and similar amino acids are indicated by black and grey shading, respectively. The underlined sequence indicates the conserved ovate domain. (B) Phylogenetic analysis. The deduced amino acid sequences were aligned using Clustal W, and the dendrogram was drawn with the neighbor-joining method using MEGA software (Arizona State University, Tempe, AZ, USA). The number for each interior branch is the bootstrap percentage calculated from 1,000 replicates. The scale bar corresponds to 0.2 amino acid substitutions per residue. The protein sequences of the OFPs used for construction of the tree are listed in the GenBank database under the following locus or accession numbers: MaAchr7 (GSMUA_Achr5T17440_001), MaAchr5 (GSMUA_ Achr5 T17440_001), MaAchr6 (GSMUA_Achr6T07060_001), MaAchr9 (GSMUA_ Achr9T02650_ 001), AtOFP1 (Arabidopsis thaliana, NP_ 195804), AtOFP2 (NP_ 180599), AtOFP3 (NP_ 200644), AtOFP4 (NP_ 172174), AtOFP5 (NP_ 193618), AtOFP6 (NP_ 680125), AtOFP7 (NP_ 179440), AtOFP8 (NP_ 197466), AtOFP9 (NP_ 192312), AtOFP10 (NP_ 197616), AtOFP11 (NP_ 193222), AtOFP12 (NP_ 172033), AtOFP13 (NP_ 196102), AtOFP14 (NP_ 178114), AtOFP15 (NP_ 565833), AtOFP16 (NP_ 180770), AtOFP17 (NP_ 850144), AtOFP18 (NP_ 566967).
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pone.0123870.g001: Sequence analysis of MaOFP1.(A) Sequence alignments of MaOFP1 with four other OFPs from the banana A genome. MaAchr7, 5, 6 and 9 represent OFPs located on chromosomes 7, 5, 6 and 9, respectively. Identical and similar amino acids are indicated by black and grey shading, respectively. The underlined sequence indicates the conserved ovate domain. (B) Phylogenetic analysis. The deduced amino acid sequences were aligned using Clustal W, and the dendrogram was drawn with the neighbor-joining method using MEGA software (Arizona State University, Tempe, AZ, USA). The number for each interior branch is the bootstrap percentage calculated from 1,000 replicates. The scale bar corresponds to 0.2 amino acid substitutions per residue. The protein sequences of the OFPs used for construction of the tree are listed in the GenBank database under the following locus or accession numbers: MaAchr7 (GSMUA_Achr5T17440_001), MaAchr5 (GSMUA_ Achr5 T17440_001), MaAchr6 (GSMUA_Achr6T07060_001), MaAchr9 (GSMUA_ Achr9T02650_ 001), AtOFP1 (Arabidopsis thaliana, NP_ 195804), AtOFP2 (NP_ 180599), AtOFP3 (NP_ 200644), AtOFP4 (NP_ 172174), AtOFP5 (NP_ 193618), AtOFP6 (NP_ 680125), AtOFP7 (NP_ 179440), AtOFP8 (NP_ 197466), AtOFP9 (NP_ 192312), AtOFP10 (NP_ 197616), AtOFP11 (NP_ 193222), AtOFP12 (NP_ 172033), AtOFP13 (NP_ 196102), AtOFP14 (NP_ 178114), AtOFP15 (NP_ 565833), AtOFP16 (NP_ 180770), AtOFP17 (NP_ 850144), AtOFP18 (NP_ 566967).

Mentions: A Y2H assay was performed using MuMADS1 as bait and 2 day postharvest (DPH) banana fruit cDNA library as prey. After identification on different minimal synthetic dropout (SD) supplements media, plasmids from the blue clones were isolated. Through Y2H we obtained a full-length ovate family protein gene designated MaOFP1, which contained a 542 bp sequence with a 438 bp full-length open reading frame (ORF) that encodes a protein of 146 amino acids. Bioinformatics analysis showed that MaOFP1 has a well-conserved ovate domain in its C-terminus and shared 37.91% identity with four other ovate family protein genes from the banana A genome that are located on chromosomes 7, 5, 6 and 9 (Fig 1A). Phylogenetic analysis showed that MaOFP1 was most closely linked with AtOFP18, and then AtOFP6 and AtOFP10 (Fig 1B). MaOFP1 showed a distant relationship with four other ovate family proteins from the banana A genome [33].


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)

Sequence analysis of MaOFP1.(A) Sequence alignments of MaOFP1 with four other OFPs from the banana A genome. MaAchr7, 5, 6 and 9 represent OFPs located on chromosomes 7, 5, 6 and 9, respectively. Identical and similar amino acids are indicated by black and grey shading, respectively. The underlined sequence indicates the conserved ovate domain. (B) Phylogenetic analysis. The deduced amino acid sequences were aligned using Clustal W, and the dendrogram was drawn with the neighbor-joining method using MEGA software (Arizona State University, Tempe, AZ, USA). The number for each interior branch is the bootstrap percentage calculated from 1,000 replicates. The scale bar corresponds to 0.2 amino acid substitutions per residue. The protein sequences of the OFPs used for construction of the tree are listed in the GenBank database under the following locus or accession numbers: MaAchr7 (GSMUA_Achr5T17440_001), MaAchr5 (GSMUA_ Achr5 T17440_001), MaAchr6 (GSMUA_Achr6T07060_001), MaAchr9 (GSMUA_ Achr9T02650_ 001), AtOFP1 (Arabidopsis thaliana, NP_ 195804), AtOFP2 (NP_ 180599), AtOFP3 (NP_ 200644), AtOFP4 (NP_ 172174), AtOFP5 (NP_ 193618), AtOFP6 (NP_ 680125), AtOFP7 (NP_ 179440), AtOFP8 (NP_ 197466), AtOFP9 (NP_ 192312), AtOFP10 (NP_ 197616), AtOFP11 (NP_ 193222), AtOFP12 (NP_ 172033), AtOFP13 (NP_ 196102), AtOFP14 (NP_ 178114), AtOFP15 (NP_ 565833), AtOFP16 (NP_ 180770), AtOFP17 (NP_ 850144), AtOFP18 (NP_ 566967).
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pone.0123870.g001: Sequence analysis of MaOFP1.(A) Sequence alignments of MaOFP1 with four other OFPs from the banana A genome. MaAchr7, 5, 6 and 9 represent OFPs located on chromosomes 7, 5, 6 and 9, respectively. Identical and similar amino acids are indicated by black and grey shading, respectively. The underlined sequence indicates the conserved ovate domain. (B) Phylogenetic analysis. The deduced amino acid sequences were aligned using Clustal W, and the dendrogram was drawn with the neighbor-joining method using MEGA software (Arizona State University, Tempe, AZ, USA). The number for each interior branch is the bootstrap percentage calculated from 1,000 replicates. The scale bar corresponds to 0.2 amino acid substitutions per residue. The protein sequences of the OFPs used for construction of the tree are listed in the GenBank database under the following locus or accession numbers: MaAchr7 (GSMUA_Achr5T17440_001), MaAchr5 (GSMUA_ Achr5 T17440_001), MaAchr6 (GSMUA_Achr6T07060_001), MaAchr9 (GSMUA_ Achr9T02650_ 001), AtOFP1 (Arabidopsis thaliana, NP_ 195804), AtOFP2 (NP_ 180599), AtOFP3 (NP_ 200644), AtOFP4 (NP_ 172174), AtOFP5 (NP_ 193618), AtOFP6 (NP_ 680125), AtOFP7 (NP_ 179440), AtOFP8 (NP_ 197466), AtOFP9 (NP_ 192312), AtOFP10 (NP_ 197616), AtOFP11 (NP_ 193222), AtOFP12 (NP_ 172033), AtOFP13 (NP_ 196102), AtOFP14 (NP_ 178114), AtOFP15 (NP_ 565833), AtOFP16 (NP_ 180770), AtOFP17 (NP_ 850144), AtOFP18 (NP_ 566967).
Mentions: A Y2H assay was performed using MuMADS1 as bait and 2 day postharvest (DPH) banana fruit cDNA library as prey. After identification on different minimal synthetic dropout (SD) supplements media, plasmids from the blue clones were isolated. Through Y2H we obtained a full-length ovate family protein gene designated MaOFP1, which contained a 542 bp sequence with a 438 bp full-length open reading frame (ORF) that encodes a protein of 146 amino acids. Bioinformatics analysis showed that MaOFP1 has a well-conserved ovate domain in its C-terminus and shared 37.91% identity with four other ovate family protein genes from the banana A genome that are located on chromosomes 7, 5, 6 and 9 (Fig 1A). Phylogenetic analysis showed that MaOFP1 was most closely linked with AtOFP18, and then AtOFP6 and AtOFP10 (Fig 1B). MaOFP1 showed a distant relationship with four other ovate family proteins from the banana A genome [33].

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