title: Successful expression of a novel bacterial gene for pinoresinol reductase and its effect on lignan biosynthesis in transgenic Arabidopsis thaliana.
abstract: Pinoresinol reductase and pinoresinol/lariciresinol reductase play important roles in an early step of lignan biosynthesis in plants. The activities of both enzymes have also been detected in bacteria. In this study, pinZ, which was first isolated as a gene for bacterial pinoresinol reductase, was constitutively expressed in Arabidopsis thaliana under the control of the cauliflower mosaic virus 35S promoter. Higher reductive activity toward pinoresinol was detected in the resultant transgenic plants but not in wild-type plant. Principal component analysis of data from untargeted metabolome analyses of stem, root, and leaf extracts of the wild-type and two independent transgenic lines indicate that pinZ expression caused dynamic metabolic changes in stems, but not in roots and leaves. The metabolome data also suggest that expression of pinZ influenced the metabolisms of lignan and glucosinolates but not so much of neolignans such as guaiacylglycerol-8-O-4'-feruloyl ethers. In-depth quantitative analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) indicated that amounts of pinoresinol and its glucoside form were markedly reduced in the transgenic plant, whereas the amounts of glucoside form of secoisolariciresinol in transgenic roots, leaves, and stems increased. The detected levels of lariciresinol in the transgenic plant following β-glucosidase treatment also tended to be higher than those in the wild-type plant. Our findings indicate that overexpression of pinZ induces change in lignan compositions and has a major effect not only on lignan biosynthesis but also on biosynthesis of other primary and secondary metabolites.
authors: Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan.
authors: Masayuki
authors: M
authors: Tamura
authors: Yukiko
authors: Y
authors: Tsuji
authors: Tatsuya
authors: T
authors: Kusunose
authors: Atsushi
authors: A
authors: Okazawa
authors: Naofumi
authors: N
authors: Kamimura
authors: Tetsuya
authors: T
authors: Mori
authors: Ryo
authors: R
authors: Nakabayashi
authors: Shojiro
authors: S
authors: Hishiyama
authors: Yuki
authors: Y
authors: Fukuhara
authors: Hirofumi
authors: H
authors: Hara
authors: Kanna
authors: K
authors: Sato-Izawa
authors: Toshiya
authors: T
authors: Muranaka
authors: Kazuki
authors: K
authors: Saito
authors: Yoshihiro
authors: Y
authors: Katayama
authors: Masao
authors: M
authors: Fukuda
authors: Eiji
authors: E
authors: Masai
authors: Shinya
authors: S
authors: Kajita
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D017705:Lignans / Q000096:biosynthesis*; D060847:Metabolic Engineering; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D042301:Sphingomonadaceae / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D017705:Lignans; D010088:Oxidoreductases; C084969:pinoresinol reductase