DEVELOPING THE iPBS STRATEGY FOR YAKON GERMPLASM EVALUATION

Abstract

Two yacon varieties PER05 and ECU45 were used for iPBS method developing for yacon - Smallanthus sonchifolius, (Poepp. et Endl.) germplasm evaluation. Because of high level of polyphenols in yacon, four DNA extraction methods were tested for the best results in the iPBS method. Using a set of universal primers that anneal to the conserved regions of retrotransposons, polymorphism of amplified fragments of DNA was analysed and for the development of iPBS protocol primers that produce PCR fragments within the whole possible range of PCR were chosen. Selected primers were subsequently used in a set of gradient PCR for finding of optimal annealing temperatures for each of them and three groups of primers according to the optimal annealing temperature were found - primers with a optimum at 53°C (1845, 1875 and 1886), at 56°C (1846) and 61 °C (1880 and 2078).