title: Changes in cinnamyl alcohol dehydrogenase activities from sugarcane cultivars inoculated with Sporisorium scitamineum sporidia.
abstract: This study describes a method for determining cinnamyl alcohol dehydrogenase activity in sugarcane stems using reverse phase (RP) high-performance liquid chromatography to elucidate their possible lignin origin. Activity is assayed using the reverse mode, the oxidation of hydroxycinnamyl alcohols into hydroxycinnamyl aldehydes. Appearance of the reaction products, coniferaldehyde and sinapaldehyde is determined by measuring absorbance at 340 and 345 nm, respectively. Disappearance of substrates, coniferyl alcohol and sinapyl alcohol is measured at 263 and 273 nm, respectively. Isocratic elution with acetonitrile:acetic acid through an RP Mediterranea sea C18 column is performed. As case examples, we have examined two different cultivars of sugarcane; My 5514 is resistant to smut, whereas B 42231 is susceptible to the pathogen. Inoculation of sugarcane stems elicits lignification and produces significant increases of coniferyl alcohol dehydrogenase (CAD) and sinapyl alcohol dehydrogenase (SAD). Production of lignin increases about 29% in the resistant cultivar and only 13% in the susceptible cultivar after inoculation compared to uninoculated plants. Our results show that the resistance of My 5514 to smut is likely derived, at least in part, to a marked increase of lignin concentration by the activation of CAD and SAD.
authors.affiliation: Department of Plant Biology I (Plant Physiology), Faculty of Biology, Complutense University, 12 José Antonio Novais Av., Madrid 28040, Spain.
authors.forename: Rocío
authors.initials: R
authors.lastname: Santiago
authors.forename: Borja
authors.initials: B
authors.lastname: Alarcón
authors.forename: Roberto
authors.initials: R
authors.lastname: de Armas
authors.forename: Carlos
authors.initials: C
authors.lastname: Vicente
authors.forename: María Estrella
authors.initials: ME
authors.lastname: Legaz
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D060467:Disease Resistance / Q000502:physiology; D004789:Enzyme Activation; D014644:Genetic Variation; D005838:Genotype; D054884:Host-Pathogen Interactions; D008031:Lignin / Q000378:metabolism; D010636:Phenols / Q000378:metabolism; D010666:Phenylpropionates / Q000378:metabolism; D010935:Plant Diseases / Q000382:microbiology*; D010937:Plant Growth Regulators / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism; D031786:Saccharum / Q000201:enzymology* / Q000235:genetics / Q000382:microbiology*; D014587:Ustilaginales / Q000502:physiology*
chemical_list: D010636:Phenols; D010666:Phenylpropionates; D010937:Plant Growth Regulators; C496130:sinapyl alcohol; D008031:Lignin; C010559:coniferyl alcohol; D000429:Alcohol Oxidoreductases; C018656:cinnamyl alcohol dehydrogenase