Identification of UDP glucosyltransferases from the aluminum-resistant tree Eucalyptus camaldulensis forming β-glucogallin, the precursor of hydrolyzable tannins. (August 2018)

• Record Type: Journal Article
• Authors: Tahara, Ko; Nishiguchi, Mitsuru; Frolov, Andrej; Mittasch, Juliane; Milkowski, Carsten
• Is Part Of: Phytochemistry. Volume 152(2018)
• Page Start: 154
• Page End: 161
• Publication Date: 2018-08
• Subjects: Eucalyptus camaldulensis -- Myrtaceae -- Aluminum tolerance -- Biosynthetic pathway -- Gallotannin -- Galloylglucose -- Glucosyltransferase -- UGT -- β-Glucogallin; Botanical chemistry -- Periodicals; Biochemistry -- Periodicals; Botany -- Periodicals; Chimie végétale -- Périodiques; 572.2
• DOI: 10.1016/j.phytochem.2018.05.005
• Languages: English
• ISSNs: 0031-9422

Abstract

Abstract: In the highly aluminum-resistant tree Eucalyptus camaldulensis, hydrolyzable tannins are proposed to play a role in internal detoxification of aluminum, which is a major factor inhibiting plant growth on acid soils. To understand and modulate the molecular mechanisms of aluminum detoxification by hydrolyzable tannins, the biosynthetic genes need to be identified. In this study, we identified and characterized genes encoding UDP-glucose:gallate glucosyltransferase, which catalyzes the formation of 1- O -galloyl-β-d -glucose (β-glucogallin), the precursor of hydrolyzable tannins. By homology-based cloning, seven full-length candidate cDNAs were isolated from E. camaldulensis and expressed in Escherichia coli as recombinant N-terminal His-tagged proteins. Phylogenetic analysis classified four of these as UDP glycosyltransferase (UGT) 84A subfamily proteins (UGT84A25a, -b, UGT84A26a, -b) and the other three as UGT84J subfamily proteins (UGT84J3, -4, -5). In vitro enzyme assays showed that the UGT84A proteins catalyzed esterification of UDP–glucose and gallic acid to form 1- O -galloyl-β-d -glucose, whereas the UGT84J proteins were inactive. Further analyses with UGT84A25a and −26a indicated that they also formed 1- O -glucose esters of other structurally related hydroxybenzoic and hydroxycinnamic acids with a preference for hydroxybenzoic acids. The UGT84A genes were expressed in leaves, stems, and roots of E. camaldulensis, regardless of aluminum stress. Taken together, our results suggest that the UGT84A subfamily enzymes of E. camaldulensis are responsible for constitutive production of 1- O -galloyl-β-d -glucose, which is the first step of hydrolyzable tannin biosynthesis. Graphical abstract: Image 1 Highlights: Seven cDNAs of UDP glycosyltransferase were isolated from Eucalyptus camaldulensis. Enzymes were named UGT84A25a, -A25b, -A26a, -A26b, -J3, -J4, and -J5, respectively. UGT84A25/26 enzymes catalyzed 1- O -galloyl-β-d -glucose (β-glucogallin) formation. UGT84A25/26 may provide precursors of hydrolyzable tannins involved in Al resistance. The catalytic activity of UGT84J3-5 remains to be elucidated.