Easy PCR method to isolate unknown ACC synthase genes in ornamental plant species.

摘要

Ornamental plants represent a high number of species from different plant families. To investigate the ethylene biosynthesis and isolate the genes involved in each species, a fast and simple method is necessary. The enzyme that controls the ethylene synthesis is the 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, which catalyses the formation of ACC, the immediate precursor of ethylene. Cloning of the first ACC synthase genes from different plant species revealed a multi-gene family. By using degenerate oligonucleotides that correspond to conserved amino acid residues of known ACC synthases, we amplified different genomic fragments that encodes ACC synthase homologue DNA fragments. As a result of the intron size variation of ACC synthase genes, the amplification of several ACC synthase genes with only one PCR reaction was possible. By means of this method, we were able to isolate several unknown ACC synthase genes in a wide range of ornamental plant species such as Rosa hybrida, Euphorbia pulcherrima, Kalanchoe blossfeldiana, Pelargonium zonale and Campanula carpatica..