Seleção de primers para análise de Inter Simple Sequence Repeats na cultivar 'Itália' de Vitis vinífera L.

Abstract

The objective of present study was to select ISSR (Inter-Simple Sequence Repeats) primers suitable for future genetic variability studies of the grape cultivar 'Itália' (Vitis vinifera L.) plants. Besides, it is aimed to standardize the method for quantifying DNA and PCR reaction for the selected primers. For DNA extraction, samples from young vine leaves collected in the region of Marialva-PR were used, and the chosen DNA quantification method was Picodrop® spectrophotometer. The primers selected for presenting a satisfactory number of sharp bands (106 in total) in 2% agarose gel when subjected to electrophoresis were: ISSR-1, ISSR-2, ISSR-5, ISSR-6, ISSR-7, ISSR-8, ISSR-9, ISSR-11, ISSR-12, ISSR-13, ISSR-14 and ISSR-15 using an annealing temperature of 50°C at the PCR. The average number of bands per primer was 8.84, whereas ISSR-5 primer produced the highest number (13) of amplified ISSR regions.