Distinction of Araucaria angustifolia seeds from different locations in Brazil by a specific DNA sequence

Brazil pine (Araucaria angustifolia [Bert.] O. Ktze.) used to be a forest-d ominating tree in areas above 500 m between 18 degrees and 30 degrees of so uthern latitude in Brazil. Due to severe clear cutting in the nineteenth an d twentieth century, only small local remnants of this landscape-forming co nifer are left. Larger attempts at reforestation of grassland areas with th is tree in the original region of occurrence have failed so far. Of the pos sible reasons, use of less suitable provenances (ecotypes) could be involve d. It was thus the aim of this investigation to screen for DNA sequences th at could possibly be used to identify such ecotypes. For this purpose, seed s from all relevant remaining natural Brazil pine locations in Brazil were collected. Using repetitive oligonucleotide primers, we were able to identi fy a DNA fragment of 2 kbp length. Interestingly, the frequency of this fra gment occurs in parallel with the number, duration and strength of frost pe riods. Although not a typical microsatellite sequence, this fragment could be used as a marker to screen for ecotypes of Brazil pine. Up to now, we ha ve not been able to find homologies for this fragment on the basis of the n ucleotide and the amino acid sequences in the respective libraries.