title: An Intracellular Laccase Is Responsible for Epicatechin-Mediated Anthocyanin Degradation in Litchi Fruit Pericarp.
abstract: In contrast to the detailed molecular knowledge available on anthocyanin synthesis, little is known about its catabolism in plants. Litchi (Litchi chinensis) fruit lose their attractive red color soon after harvest. The mechanism leading to quick degradation of anthocyanins in the pericarp is not well understood. An anthocyanin degradation enzyme (ADE) was purified to homogeneity by sequential column chromatography, using partially purified anthocyanins from litchi pericarp as a substrate. The purified ADE, of 116 kD by urea SDS-PAGE, was identified as a laccase (ADE/LAC). The full-length complementary DNA encoding ADE/LAC was obtained, and a polyclonal antibody raised against a deduced peptide of the gene recognized the ADE protein. The anthocyanin degradation function of the gene was confirmed by its transient expression in tobacco (Nicotiana benthamiana) leaves. The highest ADE/LAC transcript abundance was in the pericarp in comparison with other tissues, and was about 1,000-fold higher than the polyphenol oxidase gene in the pericarp. Epicatechin was found to be the favorable substrate for the ADE/LAC. The dependence of anthocyanin degradation by the enzyme on the presence of epicatechin suggests an ADE/LAC epicatechin-coupled oxidation model. This model was supported by a dramatic decrease in epicatechin content in the pericarp parallel to anthocyanin degradation. Immunogold labeling transmission electron microscopy suggested that ADE/LAC is located mainly in the vacuole, with essential phenolic substances. ADE/LAC vacuolar localization, high expression levels in the pericarp, and high epicatechin-dependent anthocyanin degradation support its central role in pigment breakdown during pericarp browning.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Fang
authors.initials: F
authors.lastname: Fang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Xue-lian
authors.initials: XL
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Hong-hui
authors.identifier: http://orcid.org/0000-0003-1861-0651
authors.initials: HH
authors.lastname: Luo
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Jia-jian
authors.identifier: http://orcid.org/0000-0001-5206-0791
authors.initials: JJ
authors.lastname: Zhou
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Yi-hui
authors.initials: YH
authors.lastname: Gong
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Wen-jun
authors.initials: WJ
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Zhao-wan
authors.initials: ZW
authors.lastname: Shi
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Quan
authors.initials: Q
authors.lastname: He
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Qing
authors.initials: Q
authors.lastname: Wu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Lu
authors.initials: L
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Lin-lin
authors.initials: LL
authors.lastname: Jiang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Zhi-gao
authors.initials: ZG
authors.lastname: Cai
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.).
authors.forename: Michal
authors.initials: M
authors.lastname: Oren-Shamir
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.) xqpang@scau.edu.cn zqzhang@scau.edu.cn.
authors.forename: Zhao-qi
authors.initials: ZQ
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources (F.F., X.Z., H.L., J.Z., Y.G., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., Z.Z., X.P.), College of Life Sciences (F.F., X.Z., H.L., J.Z., W.L., Z.S., Q.H., Q.W., L.L., L.J., Z.C., X.P.), and College of Horticulture (Y.G., Z.Z.), South China Agricultural University, Guangzhou 510642, China;Research Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China (F.F.); andDepartment of Ornamental Horticulture, Agriculture Research Organization, Volcani Centre, Bet Dagan 50250, Israel (M.O.-S.) xqpang@scau.edu.cn zqzhang@scau.edu.cn.
authors.forename: Xue-qun
authors.identifier: http://orcid.org/0000-0003-4522-2152
authors.initials: XQ
authors.lastname: Pang
mesh_terms: D000872:Anthocyanins / Q000378:metabolism*; D002392:Catechin / Q000378:metabolism*; D004156:Catechol Oxidase / Q000378:metabolism; D005638:Fruit / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D042845:Laccase / Q000235:genetics / Q000378:metabolism*; D032125:Litchi / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010636:Phenols / Q000378:metabolism; D010802:Phylogeny; D018515:Plant Leaves / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000502:physiology
chemical_list: D000872:Anthocyanins; D010636:Phenols; D010940:Plant Proteins; D002392:Catechin; D004156:Catechol Oxidase; D042845:Laccase