Application of genomic DNA and RAPD-PCR in genetic analysis and fingerprinting of various species of woody trees

Abstract

Summary Positive identification of tree species, subspecies and accessions is often difficult or impossible based primarily on morphological traits, and a technique ensuring reliable, repeatable and unique species identification is required. The use of molecular markers offers such a technique, allowing assessment of polymorphic variation among and between species directly at the DNA level. In this study we have applied the technique of random amplification of polymorphic DNA using arbitrary primers (RAPD-PCR) to analyse the relationship between four accessions of E. microcorys, and their differences to E. dalrympleana and Grevillea ‘Elegance’ Additionally the versatility of RAPD-PCR was demonstrated by determining genetic differences between the bark and variously aged wood samples in two Robinia pseudoacacia trees. It was necessary to test a small number of commercially available ten-base synthetic oligonucleotide primers before arriving at one which clearly distinguished all accessions, species and genera tested. The variability of the technique, and in particular the origin and quality of the DNA extracted was analysed. We demonstrated that the procedures and protocols developed are applicable to all tissue types tested from isolated protoplasts, to micropropagated shoots, to leaf samples from seedlings, and bark and wood of mature trees. Our results show that RAPD technology is a quick, precise, sensitive and a relatively inexpensive technique for genomic analysis of trees.