Application of molecular markers in the hybrid verification and assessment of somaclonal variation from oil palm propagated in vitro

摘要

Randomly amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) were used to assess the hybrid verification and somaclonal variation of the cross 366 (D) x 72 (P) of Elaeis quineensis Jacq. DxP. DNA from mature zygotic embryo (MZE) was isolated and detected by seven primers of RAPD and eight primers of SSR. Half MZEs consisting of coleoptiles regions were cultured on Murashige and Skoog (MS) medium plus various kinds and concentrations of auxins. Variation of somaclone obtained through somatic embryogenesis was detected at the proliferation stage of the culture period. The result revealed that all random primers and the SSR primers tested could amplify parental DNA. Primer OPT06 (for RAPD) and EgCIR1772 (for SSR) provided clear DNA patterns and could be used for verifying the hybridity of the cross 366 (D) x 72 (P). The highest frequency of nodular callus formation at 87.5% was obtained on MS medium supplemented with 2.50 mg/1 of 3,6-dichloro-o-anisic acid (dicamba). This was significantly different from other kinds and concentrations of auxins. Somatic embryo line at globular stage derived from each half MZE showed no variation in RAPD banding patterns with primer OPT06. It was concluded that no somaclonal variation occurred in our propagation system by RAPD marker, no major genetic changes were observed, and this augurs well for the propagation system being employed since somaclonal variation may be minimized or absent altogether