Development of clonal propagation protocols for Uapaca kirkiana and Pappea capensis, two southern African trees with economic potential

Abstract

Experiments were carried out with the objectives of developing propagation protocols for Uapaca kirkiana and Pappea capensis tree species of southern Africa, and evaluating the graft compatibility within U. Kirkiana tree clones, provenances and species. Reverse phase high performance liquid chromatography (RP-HPLC), Folin-Ciocalteau reagent, fluorescence microscopy and callus fusion methodologies were used to diagnose graft compatibility. Results indicated that U. Kirkiana culture asepsis was achieved with 0.1% w/v mercuric chloride HgCl2) and using pre-conditioned grafted trees. Sodium hypochlorite (NaOCl) improved P. Capensis seed asepsis and germination, and discarding floating seeds improved germination. Murashige and Skoog (MS) medium with 2.0 mg l-1 benzylaminopurine (BAP) and 0.3 mg l-1 casein hydrolysate (CH) was superior in shoot multiplication and 0.5 mg l-1 indole-3-butyric acid (IBA) for rooting of P. Capensis microshoots. For somatic embryogenesis, three quarter strength MS medium with 0.05 mg l-1 thidiazuron (TDZ) and 0.3 mg l-1 CH, or 0.2 mg l-1 BAP with 0.3 mg l-1 CH, were effective in germination of P. Capensis somatic embryos. For U. Kirkiana lateral shoot explants, shoot multiplication was superior on three quarter strength MS medium with 0.1 mg l-1 BAP and 0.3 mg l-1 CH. Rooting of micro-cuttings (36%) was achieved on ½ MS with 2.5 mg l-1 IBA. RP-HPLC, fluorescence microscopy and callus fusion studies showed that phenolic compounds play a major role in U. Kirkiana graft incompatibility. Less graft compatible combinations showed an increase in phenol deposits above the union and graft incompatibility was more pronounced above the union than below the union. Proliferation of parenchymatous tissues was better below the union than above the union. Fluorescence microscopy showed presence of flavonoids and polymers above the union of less graft compatible combinations. The chromatograms showed that ferulic acid was abundant and responsible for wood discolouration. The chromatograms also isolated ρara-coumaric acids which were predominant above the union of the less compatible combinations. Therefore, ρara-coumaric acids, flavonoids and polymers were implicated in graft incompatibility of U. kirkiana trees. Copyright