DNA Fingerprinting of Tissue Culture Material by Using SSR and AFLP

Abstract

Tissue culture is used as a tool in oil palm breeding programme and commercial vegetative propagation. To control tissue culture process in a laboratory, DNA analysis can be used to verify the identity information and to understand the genetic stability of each stage of tissue culture. This research aimed to study the DNA extraction of oil palm tissue culture material and DNA fingerprinting of tissue culture material at each stage of tissue culture process, by using 11 SSR markers and 6 combinations of AFLP selective primers. The results showed that the protocol of  DNA extraction resulted DNA samples with good quality and quantity for SSR-PCR and AFLP-PCR. The same SSR profiles were observed at all material analyzed at each stage of tissue culture process. There were variations of AFLP profiles observed at material of explant and callus stages, as well as embryo and ramet stages. Different DNA profiles of the same material at different stages show genetic changes of culture material which probably caused by the process of tissue culture. SSR could be used for identification or verification of culture material identities, while the DNA markers which detected different DNA profiles could be used for further study of culture material genetic changes, in relation with clone abnormality.