title: Flavan-3-ols in Norway spruce: biosynthesis, accumulation, and function in response to attack by the bark beetle-associated fungus Ceratocystis polonica.
abstract: Proanthocyanidins (PAs) are common polyphenolic polymers of plants found in foliage, fruit, bark, roots, rhizomes, and seed coats that consist of flavan-3-ol units such as 2,3-trans-(+)-catechin and 2,3-cis-(-)-epicatechin. Although the biosynthesis of flavan-3-ols has been studied in angiosperms, little is known about their biosynthesis and ecological roles in gymnosperms. In this study, the genes encoding leucoanthocyanidin reductase, a branch point enzyme involved in the biosynthesis of 2,3-trans-(+)-flavan-3-ols, were identified and functionally characterized in Norway spruce (Picea abies), the most widespread and economically important conifer in Europe. In addition, the accumulation of flavan-3-ols and PAs was investigated in Norway spruce saplings after wounding or inoculation with the fungal pathogen Ceratocystis polonica, which is vectored by bark beetles (Ips typographus) and is usually present during fatal beetle attacks. Monomeric and dimeric flavan-3-ols were analyzed by reverse-phase high-pressure liquid chromatography, while the size and subunit composition of larger PAs were characterized using a novel acid hydrolysis method and normal phase chromatography. Only flavan-3-ol monomers with 2,3-trans stereochemistry were detected in spruce bark; dimeric and larger PAs contained flavan-3-ols with both 2,3-trans and 2,3-cis stereochemistry. Levels of monomers as well as PAs with a higher degree of polymerization increased dramatically in spruce bark after infection by C. polonica. In accordance with their role in the biosynthesis of 2,3-trans-(+)-flavan-3-ols, transcript abundance of Norway spruce LEUCOANTHOCYANIDIN REDUCTASE genes also increased significantly during fungal infection. Bioassays with C. polonica revealed that the levels of 2,3-trans-(+)-catechin and PAs that are produced in the tree in response to fungal infection inhibit C. polonica growth and can therefore be considered chemical defense compounds.
authors.affiliation: Max Planck Institute for Chemical Ecology, 07745 Jena, Germany.
authors.forename: Almuth
authors.initials: A
authors.lastname: Hammerbacher
authors.forename: Christian
authors.initials: C
authors.lastname: Paetz
authors.forename: Louwrance P
authors.initials: LP
authors.lastname: Wright
authors.forename: Thilo C
authors.initials: TC
authors.lastname: Fischer
authors.forename: Joerg
authors.initials: J
authors.lastname: Bohlmann
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Davis
authors.forename: Trevor M
authors.initials: TM
authors.lastname: Fenning
authors.forename: Jonathan
authors.initials: J
authors.lastname: Gershenzon
authors.forename: Axel
authors.initials: A
authors.lastname: Schmidt
mesh_terms: D000818:Animals; D000872:Anthocyanins / Q000378:metabolism; D001203:Ascomycota / Q000254:growth & development / Q000502:physiology*; D055162:Biocatalysis; D053898:Biosynthetic Pathways / Q000235:genetics; D002392:Catechin / Q000378:metabolism; D001517:Coleoptera / Q000382:microbiology*; D005419:Flavonoids / Q000096:biosynthesis / Q000737:chemistry*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D009664:Norway; D010802:Phylogeny; D028222:Picea / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology*; D024301:Plant Bark / Q000469:parasitology*; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014158:Transcription, Genetic
chemical_list: D000872:Anthocyanins; D005419:Flavonoids; D011994:Recombinant Proteins; C404987:flavan-3-ol; D002392:Catechin