2 Directorate of Nature Conservation, Gauteng Provincial Government, PO BOX 9769, JHB 2000.

The year 2003 signalled fifty years since the discovery of the DNA double helix, and during this time interval almost no aspect of biological science has remained untouched by the ensuing revolution in molecular biology. Taxonomy and systematics have been no exception, and it is now common practice to generate DNA sequence data to assist in our understanding of evolutionary (or phylogenetic) relationships among groups of organisms. In addition DNA fingerprinting offers a means to produce unique genetic barcodes for species, an application particularly relevant to the wildlife trade. Modern molecular techniques thus represent a powerful tool to address key problems in biodiversity and conservation science, in addition to contributing to much needed capacity building in high-profile biotechnologies in South Africa.

DNA fingerprinting of critically endangered cycad species - the Gauteng Directorate of Nature Conservation of the Gauteng Department of Agriculture, Conservation, Environment and Land Affairs (GDACEL) is required to regulate the trade in cycads (genus of Encephalartos) in terms of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) and the Gauteng Nature Conservation Ordinance (Ordinance 12 of 1983).

A discussion document submitted to Working Group 1 (Conservation and Biodiversity) entitled "Cycads, their conservation and sustainable utilization in South Africa" concluded that trade in wild collected plants has been and is still the most important threat to cycad populations, with 60% of the decline in cycad numbers attributed to trade in wild collected plants. Legislation protecting these species and/or the enforcement of this legislation has evidently been unsuccessful. Although CITES regulations are theoretically considered as a significant deterrent to international trade in wild-collected plants, the implementation of this legislation is fraught with difficulties. A problem expressed by law enforcement agencies is that they are unable to prosecute suspected illegal traders, as they cannot prove that plants are of wild origin.

Currently, according to the law enforcement division of GDACEL, the most significant threat to South African cycads is the continued removal of whole plants and suckers from the wild. Microchips are easily removed and plants are "laundered" through the issuing of export permits in other provinces. Eight Critically Endangered species are especially considered at risk due to small wild populations and their popularity in trade, demonstrated by the exorbitant prices for adults of these species (up to R100 000 per metre of stem height). These species include Encephalartos cupidus, E. dyerianus, E. heenanii, E. hirsutus, E. inopinus, E. laevifolius, E. latifrons and E. middelburgensis.

Although forensic science methods are ideally suited to assessing the wild origins of plants, specific methods for cycads require urgent development in South Africa to address this problem. A feasibility study, funded by GDACEL in 2002/2003 and carried out by SANBI, investigated the application of the AFLP fingerprinting technique to cycads. This pilot project was highly successful and showed that it is feasible to use DNA fingerprinting to even verify the parentage of cycad seedlings.

Ideally, DNA fingerprint profiles should be constructed for all wild plants of the eight priority species identified above, with the proximate aim of eliminating the threat of collectors through strengthening law enforcement efforts and the ultimate aim of reintroducing cycads back into the wild. In collaboration with Mpumalanga Parks Board, GDACEL has commissioned SANBI to generate DNA fingerprints for the remaining individuals of Encephalartos middelburgensis, which occurs naturally in the two provinces. It is currently estimated that less than 120 individuals of this species remain in the wild, with future declines inevitable due to continued collector pressure and reproductive failure. A parallel study is also being carried out by SANBI, in collaboration with Eastern Cape Nature Conservation authorities, involving remaining individuals of Encephalartos latifrons. This species is represented by less than 60 plants in the wild and one of the main aims of the study is to determine the relative levels of genetic diversity remaining in the wild as compared to the 19 individuals housed in the ex situ collection at the Kirstenbosch Botanical Garden in Cape Town. Our preliminary results show that the Kirstenbosch collection represents a significant portion of the remaining genetic diversity of this species, and will therefore play a vital role in future in situ restoration efforts.