Micropropagation of loblolly pine by somatic organogenesis and RAPD analysis of regenerated plantlets

Abstract

Organogenesis was induced in callus derived from mature zygotic embryos of six families (J-56, S-1003, E-22, E-311, E-440, and Mc) of loblolly pine (Pinus taeda L.) within 24 weeks of culture. Elongation of adventitious buds was achieved on TE medium supplemented with 0.5 mg·L⁻¹ indole-3-butyric acid (IBA) and 1 mg/l 6-benzyladenine (BA). The most suitable medium for root formation proved to be TE medium supplemented with 0.5 mg·L⁻¹ IBA, 2mg·L⁻¹ BA and 0.5 mg/l gibberellic acid (GA3). 169 regenerated plantlets were transferred to a perlite: peatmoss: vermiculite (1∶1∶1) soil mixture, and 98 plantlets survived in the field. Total DNA was extracted from the needles of the regenerated plantlets of the six families of loblolly pine. Analysis of random amplified polymorphic DNA (RAPD) using 20 arbitrary oligonucleotide 10-mers, show that amplification products were monomorphic for all the plantlets of family J-56, S-1003, E-22, E-311, E-440, and Mc of loblolly pine. These results suggested that organogenesis can be used for clonal micropropagation of some families of loblolly pine.

Keywords

Pinus taeda L. / Organogenesis / Plant regeneration / Random amplified polymorphic DNA (RAPD)