The relation between recalcitrancy of a mangrove plant, Kandelia obovata, and high endogenous level of abscisic acid

Kandelia obovata Sheue, Liu & Yong sp. nov. is one of the cold tolerant mangrove plants. Some callus formation was obtained from the leaves of K. obovata in liquid medium containing 10 μM of 2,4-dichlorophenoxyacetic acid. However, recalcitrancy was found when subculturing them. Endogenous levels of gibberellins (GAs) and abscisic acid (ABA) in leaf protoplasts of K. obovata were determined using micro-scale extraction and purification steps, including thin layer chromatography and quantification by micro-bioassay or enzyme linked immunosorbent assays. Very high amounts of ABA and low activities of GAs were found in leaf protoplasts of K. obovata. Low concentrations of gibberellic acid and uniconazole-P were effective of enhancing cell enlargement in protoplast cultures. Exogenous application of ABA was inhibitory to protoplast culture. All cytokinins tested were inhibitory to both leaf and protoplast cultures. The high endogenous level of ABA is most likely the underlying cause of recalcitrancy of mangrove cultures.