title: MdACO expression during abscission: the use of 33P labeled primers in transcript quantitation.
abstract: The last step of ethylene biosynthesis in apple is catalyzed by ACO (1-aminocyclopropane 1-carboxylic acid oxidase) encoded by MdACO1 and MdACO2. Both genes were expressed during early fruit development although at different level. Later on, the ACO transcript accumulation in persisting fruitlets decreased whereas in fruitlets undergoing abscission MdACO2 transcripts remained constant and those of MdACO1dramatically increased. The expression of MdACO was assessed by a particular type of semi-quantitative PCR involving 33P labeled primers and the following electrophoresis in polyacrylamide gel. The use of either the reverse primer or the anchored oligo dT13 produced clear and elegant results. Nevertheless, the initial quantity of labeled primer appeared to be a crucial factor for obtaining a wide range titration curve.
authors.affiliation: Department of Environmental Agronomy and Crop Science, University of Padova, Viale dell'Università 16, Legnaro, Padova, 35020, Italy. vdalcin@ufl.edu
authors.forename: Valeriano
authors.initials: V
authors.lastname: Dal Cin
authors.forename: Giulio
authors.initials: G
authors.lastname: Galla
authors.forename: Angelo
authors.initials: A
authors.lastname: Ramina
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D016922:Cellular Senescence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000378:metabolism*; D018076:DNA, Complementary / Q000302:isolation & purification; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant; D027845:Malus / Q000201:enzymology* / Q000235:genetics; D010761:Phosphorus Radioisotopes; D012333:RNA, Messenger / Q000032:analysis*; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism