Final report -- covering period May 24, 1991-November 23, 1995 : molecular genetic analysis of valuable native timber species

tropical forest ecologist, and a premier genetic analysis laboratory in the United States to establish the foundation for a long-term program to assess levels and distribution of genetic variability within Costa Rica"s populations of economically important native timber trees. Goals were to establish a plant genetics laboratory facility at the University of Costa Rica, to develop and test molecular techniques for assessing the genetic structure of Costa Rican native timber tree species, and to carry out pilot studies with these techniques on three populations of native timber species, Minquartia guianensis, Hyeronima alchorneoides and Simarouba amara. Procedures were developed for analysis of genetic variation using two techniques: isozyme expression variation and DNA fingerprinting. This final report presents extensive results on the isozyme analysis of Minquartia guianensis populations and preliminary results on the other two species. DNA extraction procedures and the standardization of the RAPD fingerprinting are reported. Key issues were the development of a simple and reliable technique for the extraction of DNA from tree samples suitable for PCR amplification and the study of the optimum conditions for PCR amplification, where repeatability of results is paramount for the analysis. The project demonstrated the importance of collaborative efforts involving forest ecologists, molecular biologists, and population geneticists. A Costa Rican laboratory is now fully equipped and operational for the analysis of genetic variability on plant populations, and local personnel were trained in leading laboratories in the United States in genetic analysis techniques and in the optimization of DNA fingerprinting techniques using PCR. (Author abstract)