G. Paglia et M. Morgante, PCR-BASED MULTIPLEX DNA-FINGERPRINTING TECHNIQUES FOR THE ANALYSIS OFCONIFER GENOMES, Molecular breeding, 4(2), 1998, pp. 173-177
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.