title: Molecular cloning and functional characterization of DkMATE1 involved in proanthocyanidin precursor transport in persimmon (Diospyros kaki Thunb.) fruit.
abstract: Persimmon fruits accumulate a large amount of proanthocyanidins (PAs) in "tannin cells" during development that cause the sensation of astringency due to coagulation of oral proteins. Pollination-constant non-astringent (PCNA) is a spontaneous mutant persimmon phenotype that loses its astringency naturally on the tree at maturity; while the more common non-PCNA fruits remain rich in PAs until they are fully ripened. Here, we isolated a DkMATE1 gene encoding a Multidrug And Toxic Compound Extrusion (MATE) family protein from the Chinese PCNA (C-PCNA) 'Eshi 1'. Expression patterns of DkMATE1 were positively correlated with the accumulation of PAs in different types of persimmons fruits during fruit development. An analysis of the inferred amino acid sequences and phylogenetic relationships indicated that DkMATE1 is a putative PA precursor transporter, and subcellular localization assays revealed that DkMATE1 is localized in the vacuolar membrane. Ectopic expression of the DkMATE1 in Arabidopsis tt12 mutant supported that DkMATE1 could complement its biological function in transporting epicatechin 3'-O-glucoside as a PAs precursor from the cytoplasm to vacuole. Furthermore, the transient over-expression and silencing of DkMATE1 in 'Mopanshi' persimmon leaves resulted in a significant increase and a decrease in PA content, respectively. The analysis of cis-elements in DkMATE1 promoter regions indicated that DkMATE1 might be regulated by DkMYB4, another well-known structural gene in persimmon. Overall, our results show that DkMATE1 may be an essential PA precursor membrane transporter that plays an important role in PA biosynthesis in persimmon.
authors.affiliation: Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.
authors.forename: Sichao
authors.initials: S
authors.lastname: Yang
authors.affiliation: Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.
authors.forename: Liqing
authors.initials: L
authors.lastname: Xu
authors.affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Shiratake
authors.affiliation: Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Wuhan 430070, Hubei, China; Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains, Huanggang 438000, Hubei, China.
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Luo
authors.affiliation: Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University, Wuhan 430070, Hubei, China; Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains, Huanggang 438000, Hubei, China. Electronic address: zhangqinglin@mail.hzau.edu.cn.
authors.forename: Qinglin
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics; D001692:Biological Transport; D003001:Cloning, Molecular; D029788:Diospyros / Q000235:genetics / Q000378:metabolism*; D005638:Fruit / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D007425:Intracellular Membranes / Q000378:metabolism; D009154:Mutation; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D030821:Plants, Genetically Modified; D044945:Proanthocyanidins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D014157:Transcription Factors / Q000235:genetics; D014617:Vacuoles / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D010940:Plant Proteins; D044945:Proanthocyanidins; D014157:Transcription Factors; C436245:tt12 protein, Arabidopsis; C013221:proanthocyanidin
keywords: MATE; Persimmon; Proanthocyanidin; Transporter; Vacuolar membrane