title: Genetic analysis of cinnamyl alcohol dehydrogenase in loblolly pine: single gene inheritance, molecular characterization and evolution.
abstract: The gene encoding the monolignol biosynthetic enzyme cinnamyl alcohol dehydrogenase (CAD, E.C. 1.1.1.195) can be expressed in response to different developmental and environmental cues. Control of Cad gene expression could involve either differential regulation of more than one Cad gene or, alternatively combinatorial regulation of a single Cad gene. In loblolly pine (Pinus taeda L.), we found several electrophoretic variants (allozymes) of CAD and a high level of heterozygosity (he = 0.46). Analysis of inheritance patterns of pine CAD allozymes gave segregation ratios that were consistent with Mendelian expectations for a single functional gene. The identity of the full-length Cad cDNA sequence was confirmed by alignment with peptide sequences obtained from purified active enzyme and by extensive similarity to Cad sequences from other species. Southern blot analysis of genomic DNA using the Cad cDNA as a hybridization probe gave simple patterns, consistent with our interpretation that pine Cad is a single-copy gene. Phylogenetic analysis and evolution rate estimates showed that Cad sequences are diverging less rapidly in the gymnosperms than in the angiosperms. The Cad mRNA was present in both lignifying tissues and a non lignifying tissue (the megagametophyte) of pine. The presence of a single gene suggests that different regulatory mechanisms for a single Cad gene, rather than differential regulation of several genes, can account for its expression in response to different cues.
authors.affiliation: Department of Genetics, North Carolina State University, Raleigh 27695-8008, USA.
authors.forename: J J
authors.initials: JJ
authors.lastname: MacKay
authors.forename: W
authors.initials: W
authors.lastname: Liu
authors.forename: R
authors.initials: R
authors.lastname: Whetten
authors.forename: R R
authors.initials: RR
authors.lastname: Sederoff
authors.forename: D M
authors.initials: DM
authors.lastname: O'Malley
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D005075:Biological Evolution; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D041603:Pinus taeda; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D017421:Sequence Analysis
chemical_list: D007527:Isoenzymes; D000429:Alcohol Oxidoreductases; C018656:cinnamyl alcohol dehydrogenase