DNA quality is still crucial in genetic analysis of plants-Experiences with Araucaria araucana in phylogeographic studies

C. BAIER; C. MENGEL; C. MORENO; P. MARCHELLI; L. GALLO; B. ZIEGENHAGEN

Regensburg, Alemania

Simposio; Annual Meeting of the Gesellschaft fuer Oekologie; 2005

Araucaria araucana (Araucariaceae) (Molina) Koch is a species endemic to the southern South American temperate forest that grows in a limited area between 37°20´ and 40°0´. Today the species is in danger of extinction because fire and  anthropogenic use caused strong fragmentation. Therefore, the species is protected internationally by the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). The fragmentation probably caused a reduction of genetic diversity. To manage the conservation of genetic resources sustainably. it is necessary to determine the geographic distribution of intraspecific genetic variation within and between populations, thus to understand the phylogeography of this species. Phylogeographic studies commonly start with exploring plastid genomes for relevant polymorphisms. However, in this species the extracted DNA turned out to be highly contaminated by secondary metabolics.We, therefore, used a systematic approach to solve the problem by testing different methods of extraction. Furthermore, we optimised the PCR by knocking out inhibitors and designed primers to shorten the products. Single and important steps of the optimisation are demonstrated Using universal primers, we detected two candidate polymorphic chloroplast DNA regions, namely the intergenic spacer regions trnC-trnD and trnSpsbC [2]. First results are presented.