title: The accumulation and localization of chalcone synthase in grapevine (Vitis vinifera L.).
abstract: Chalcone synthase (CHS, E.C.2.3.1.74) is the first committed enzyme in the flavonoid pathway. Previous studies have primarily focused on the cloning, expression and regulation of the gene at the transcriptional level. Little is yet known about the enzyme accumulation, regulation at protein level, as well as its localization in grapevine. In present study, the accumulation, tissue and subcellular localization of CHS in different grapevine tissues (Vitis vinifera L. Cabernet Sauvignon) were investigated via the techniques of Western blotting, immunohistochemical localization, immunoelectron microscopy and confocal microscopy. The results showed that CHS were mainly accumulated in the grape berry skin, leaves, stem tips and stem phloem, correlated with flavonoids accumulation. The accumulation of CHS is developmental dependent in grape berry skin and flesh. Immunohistochemical analysis revealed that CHS were primarily localized in the exocarp and vascular bundles of the fruits during berry development; in palisade, spongy tissues and vascular bundles of the leaves; in the primary phloem and pith ray in the stems; in the growth point, leaf primordium, and young leaves of leaf buds; and in the endoderm and primary phloem of grapevine roots. Furthermore, at the subcellular level, the cell wall, cytoplasm and nucleus localized patterns of CHS were observed in the grapevine vegetative tissue cells. Results above indicated that distribution of CHS in grapevine was organ-specific and tissue-specific. This work will provide new insight for the biosynthesis and regulation of diverse flavonoid compounds in grapevine.
authors.affiliation: Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China; College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Huiling
authors.initials: H
authors.lastname: Wang
authors.affiliation: Key Laboratory of Silviculture of the State Forestry Administration, The Institute of Forestry, The Chinese Academy of Forestry, Yi He Yuan Hou, Beijing 100091, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: JiCheng
authors.initials: J
authors.lastname: Zhan
authors.affiliation: Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China), Ministry of Agriculture, China.
authors.forename: Ailing
authors.initials: A
authors.lastname: Yan
authors.affiliation: Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Sun
authors.affiliation: Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China.
authors.forename: Guojun
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China.
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China.
authors.forename: Jiancheng
authors.initials: J
authors.lastname: Ren
authors.affiliation: College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. Electronic address: huanggwd@263.net.
authors.forename: Weidong
authors.initials: W
authors.lastname: Huang
authors.affiliation: Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China. Electronic address: haiyingxu63@sina.com.
authors.forename: Haiying
authors.initials: H
authors.lastname: Xu
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D005419:Flavonoids / Q000378:metabolism; D005638:Fruit / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D007150:Immunohistochemistry; D009928:Organ Specificity; D018515:Plant Leaves / Q000201:enzymology / Q000648:ultrastructure; D018517:Plant Roots / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology; D027843:Vitis / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure
chemical_list: D005419:Flavonoids; D012333:RNA, Messenger; D000217:Acyltransferases; C020141:flavanone synthetase
keywords: Chalcone synthase; Flavonoids; Grapevine; Immunolocalization