PCR-BASED MULTIPLEX DNA-FINGERPRINTING TECHNIQUES FOR THE ANALYSIS OFCONIFER GENOMES

We utilised fingerprinting techniques such as amplified restriction fr agment polymorphism (AFLP) and selective amplification of microsatelli te polymorphic loci (SAMPL) in order to identify molecular markers in he conifer species Norway spruce (Picea abies K). A total of 366 polym orphic AFLP and 20 polymorphic SAMPL bands were scored, with an averag e of 12.6 polymorphic bands obtained per primer combination. Of the ba nds, 94% segregated in a Mendelian fashion, and 14% of the AFLP bands behaved as codominant markers. Norway spruce, as well as other conifer s, has a very large genome (1.5-2.5 x 10(10) bp) which is thought to b e largely composed of repetitive sequences of different origin, either organised in tandem or interspersed in the genome. The high complexit y of these genomes has so far hindered the efficient use of multiplexe d fingerprinting techniques for their analysis. The use of the methyla tion-sensitive restriction enzyme PstI as 6 bp cutter allows a reducti on in pattern complexity as compared to EcoRI. We thus demonstrated th at high-throughput PCR-based multiplex fingerprinting techniques such as AFLP and SAMPL, when opportunely modified, can be effectively appli ed also to these species and be useful for mapping purposes.