Construction of molecule markers for prolific oil palm tissue culture

The use of DNA marker could minimize problems in tissue culture especially when applied during the selection of plants for tissue culture. Therefore, the aimed of this research was to develop molecular markers for prolific oil palm tissue culture using amplified fragment length polymorphism (AFLP) technique. AFLP analysis was carried out upon 20 oil palm clones that have divided into three classes which are non-prolific clone (10 types of clone), normal clone (6 types of clone) and prolific clone (4 types of clone). All of the clones used were from different cell line. There were 25 primer combinations used in the AFLP analysis and 13 out of them have produced significant polymorphic amplification patterns. From these results, 44 polymorphic DNA fragments were isolated where 33 fragments for non-prolific clone, one fragment for normal clone and 10 fragments for prolific clone. These fragments were cloned into plasmid, sequenced and then sequence analysis was done. There were 36 polymorphic fragments have undergone the subsequent experiments. A pair of specific primers for each fragment was designed based on their sequences. The expected size of amplified DNA bands for each primer pair was between 70 bp to 500 bp. The optimized primer pairs were tested to the 20 types of oil palm clones in order to confirm the markers developed. From the 36 designated primers combinations, 2 pairs of the primers showed the potential to be used as marker for prolific oil palm tissue culture.