Liquid cultures were successfully generated from cotyledons of two Sonneratia species, S. alba and S. caseolaris in Murashige and Skoog (MS) medium containing 0.1 lmol L -1 2,4-dichlorophenoxyacetic acid (2,4-D). Adventitious roots differentiated from cotyledons of S. alba. Proliferated cells were subcultured and a large volume of suspension cells was subsequently established in 100-mL flasks. All the cytokinins tested inhibited cell proliferation. After three years of culture, the potential to differentiate was tested as indicated by greening of the cells. Greening occurred when suspension cells were transferred to solid MS medium with and without 0.1 lmol L -1 2,4-D. Greening was stimulated by low concentrations of the weak auxins indolebutyric acid (IBA) and naphthaleneacetic acid (NAA) while 2,4-D stimulated late-stage greening. Abscisic acid (ABA) inhibited greening. Gibberellic acid (GA 3 ) at 1.0 lmol L -1 stimulated callus greening and was not inhibitory even when tested at high concentrations. Cytokinins were inhibitory in combination with 0.1 lmol L -1 of either IBA or NAA. The cause of different effects of plant hormones on growth and differentiation was discussed. Small-scale liquid media and 24-well culture plates of solid media methods developed in this paper are suitable for the optimization of hormonal conditions for cell proliferation and differentiation.
Growing interest in ecosystem restoration has recently turned the focus on tree planting, one of the most widely used restoration tools globally. Here, we study the restoration potential of tree planting in a cool-temperate forest in Shiretoko National Park, northern Japan. We used simulation modeling to investigate the long-term success of tree planting in restoring biodiversity and the climate change mitigation function relative to intact natural forests. Specifically, we investigated 31 different restoration scenarios, consisting of five planting densities (1,000-10,000 trees/ha) Â six levels of planted tree species richness (one to six species) + one no-planting scenario. We examined these scenarios at different distances from natural forests serving as a seed source (0-300 m) to quantify the potential for natural regeneration. In restoration areas in close proximity to a natural forest, species-rich highdensity planting scenario performed best, reaching >50% of the reference values from intact natural forests within 33 years for both restoration goals. However, variation in restoration outcomes was small when >2,500 trees/ha of more than four species were planted, regardless of distance to seed source. In contrast, biodiversity restoration was considerably delayed in scenarios where planted species richness was low as well as in restoration areas that were far from a seed source yet relied solely on natural regeneration. We here demonstrate how forest landscape simulation can be used to identify viable restoration options for managers across multiple restoration goals as an important step to bridge the research-implementation gap in forest restoration.
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