Hunting monolignol transporter(s): membrane proteomics and biochemical transport assays with membrane vesicles of Norway spruce
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ABSTRACT: Monolignol transport during lignification is a partially solved puzzle: both the mechanism(s) and the transported form of monolignols are unknown in developing xylem of trees. We tested a hypothesis of an active, plasma membrane (PM)-localized transport of monolignol monomers, dimers, and/or glucosidic forms with membrane vesicles prepared of developing xylem, phloem, and lignin-forming, tissue-cultured cells of Norway spruce (Picea abies L. Karst.), as well as of a control material, non-lignifying tobacco (Nicotiana tabacum L.) BY-2 cells. Xylem and BY-2 vesicles transported both coniferin and p-coumaryl alcohol glucoside, but inhibitor assays suggested this transport being over the tonoplast. Based on similar inhibitor assays, lignin-forming, tissue42 cultured cells of spruce had coniferin transport putatively localizing on PM. Liquid chromatography-mass spectrometry (LC-MS/MS) analysis of membrane proteins isolated from spruce developing xylem, phloem and tissue-cultured cells revealed multiple transporters. These were compared to a transporter gene set that was gained by a correlation analysis with a selected set of spruce monolignol biosynthesis genes. Biochemical membrane vesicle assays showed no support for the ABC-transporter-mediated monolignol transport but point to secondary active transporters (such as MFS- or MATE-transporters). In contrast, proteomic and co-expression analyses suggest a role for ABC-transporters and MFS-transporters.
INSTRUMENT(S): Synapt G2-S HDMS
ORGANISM(S): Picea Abies
TISSUE(S): Wood
DISEASE(S): Not Available
SUBMITTER: Joakim Bygdell
LAB HEAD: Anna Kärkönen
PROVIDER: PXD017533 | Pride | 2020-08-14
REPOSITORIES: Pride
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