AFLP Analysis of Opium Poppy

Abstract

Amplified restriction fragment length polymorphic (AFLP) DNA analysis was performed on leaf samples of 40 accessions of opium poppy (Papaver somniferum L.) and two other control genera (Papaver bracteatum Lindley and Papaver setigerum DC.) from a commercial breeding collection held in Tasmania, Australia. A similarity dendrogram was produced on the basis of the analysis of all AFLP bands that ranged between 66 and 367 base pairs (bp) seen on an autoradiogram from denaturing polyacrylamide sequencing electrophoretic gels with three different primer pairs. It was necessary to combine the analysis of all three primer pairs into a single dendrogram to ensure that replicate analyses of related populations were grouped together. It was necessary to use more than one primer pair to resolve clearly the genetic relationship between the closely related plants described in this study; however, even a single primer pair could easily distinguish between populations of different poppy species. The application of AFLP DNA analysis for cultivar identification represents an efficient, reliable procedure for identifying opium poppy breeding lines in a definitive manner. This procedure required only a small amount of leaf material for analysis and the degree of the genetic relatedness was performed on autoradiographic banding patterns with commercially available software. These data were used to generate a similarity dendrogram depicting the predicted genetic relationship of the opium poppy cultivars. Opium poppy accessions sharing common parental lines could be distinguished unambiguously from poppy accessions of more distant genetic background. The banding patterns were reproducible, consistent within a genotype, and the DNA polymorphisms were frequent enough to be useful in characterizing genetic diversity in even closely related breeding lines.