RAPD Markers from Perennial Ryegrass DNA Extracted from Seeds

Abstract

As the number of perennial ryegrass (Lolium perenne L.) cultivars increases, the development of reliable identification methods becomes more important. Randomly amplified polymorphic DNA (RAPD) markers show promise in cultivar identification. Since perennial ryegrass cultivars are composites of genotypes rather than a single genotype, finding markers that distinguish cultivars is difficult. The ideal cultivar identification procedure would use seed tissue as a DNA source and evaluate a single sample as representative of a cultivar. The objective of this research was to determine whether RAPD markers could be used to consistently distinguish bulk seed samples of perennial ryegrass cultivars. Two extraction protocols were evaluated. A quick, simple extraction resulted in the amplification of few consistent RAPD markers. The more labor-intensive extraction with hexadecyltrimethyl ammonium bromide (CTAB), however, produced more reliable RAPD markers. Eight of 11 cultivars were distinguished by using RAPD markers produced using bulk seed samples together with four of 30 primers that were screened. These results show the potential of RAPD markers to provide the turfgrass industry, breeders, and certification agencies additional options to ensure the genetic integrity of perennial ryegrass seed lots and cultivars.