The aim of this project was to devise a DNA-based test for identifying natural products derived from plants at the species level. The resulting test needed to be applicable to a wide range of different species, be able to detect trace contaminants and identify pure samples of commercially processed tissues.

Degenerate PCR primers and specific PCR conditions were developed for the selective amplification of a polymorphic region of ribulose-1,5-bisphosphate carboxylase (RuBisCo) small subunit genes. Reliable amplification has been achieved for genes from more than twenty plant species from a variety of taxonomic groups. Analysis of Nicotiana species provides strong evidence that the test loci are polymorphic at the interspecies level but show little polymorphic variation at the intraspecies level. This specific multilocus PCR approach provides a powerful counterpart to RAPD and RFLP analysis for the identification of plant tissues.

Publications

Appleby, J.M. (1996). PhD Thesis, University of Leeds.

Appleby, J.M., Nelson G., McPherson, M.J. & Hamlyn, P.F. (1997). PCR amplification of the RuBisCo small subunit genes and their novel application to plant tissue identification.  Heredity, 79(6), 557-563.