title: Isolation and characterization of a chalcone isomerase gene promoter from potato cultivars.
abstract: Chalcone isomerase (CHI) is a key enzyme involved in anthocyanin metabolism. Previous research on CHI has mainly focused on cDNA cloning and gene expression. In the current study, the 1425-bp potato CHI promoter (PCP) was isolated from four potato cultivars (Heijingang, Zhongshu 7, Désirée, and Favorita) using PCR and DNA sequencing. The PCP contained many cis-regulatory elements (CREs) related to anthocyanin metabolism, tissue specificity, light response, stress, and hormone induction. Of the PCP CREs identified, 19 were common to those found in the higher plants examined, based on plant CRE databases. Multiple sequence alignment showed six single nucleotide variation sites in PCP among the potato cultivars examined, resulting in changes in the number of CREs connected with tissue specificity, anthocyanin metabolism, and light response. The 665-bp PCP fragments from Favorita and 1425-bp PCP fragments from Heijingang were used to construct plant expression vectors, which may be a useful tool for biological engineering. A transient expression assay demonstrated that the two PCP fragments from Heijingang could direct the expression of a green fluorescent protein gene in onion epidermis and a β-glucuronidase gene in all potato tuber tissues with different colors, suggesting that the single nucleotide variation in the PCP did not affect its activity, and that silencing of the CHI gene in Favorita may be attributed to other regulatory factors.
authors.affiliation: College of Life Sciences, Nanjing Agricultural University, Nanjing, China.
authors.forename: M
authors.initials: M
authors.lastname: Chen
authors.affiliation: College of Life Sciences, Nanjing Agricultural University, Nanjing, China.
authors.forename: W J
authors.initials: WJ
authors.lastname: Zhu
authors.affiliation: College of Sciences, Nanjing Agricultural University, Nanjing, China.
authors.forename: X
authors.initials: X
authors.lastname: You
authors.affiliation: College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China.
authors.forename: Y D
authors.initials: YD
authors.lastname: Liu
authors.affiliation: College of Life Sciences, Nanjing Agricultural University, Nanjing, China.
authors.forename: G M
authors.initials: GM
authors.lastname: Kaleri
authors.affiliation: College of Life Sciences, Nanjing Agricultural University, Nanjing, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Yang
mesh_terms: D000872:Anthocyanins / Q000096:biosynthesis; D001483:Base Sequence; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019753:Intramolecular Lyases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D011198:Solanum tuberosum / Q000235:genetics* / Q000378:metabolism
chemical_list: D000872:Anthocyanins; D010940:Plant Proteins; D049452:Green Fluorescent Proteins; D005966:Glucuronidase; D019753:Intramolecular Lyases; C019865:chalcone isomerase