Plant regeneration of eucalypts from rotating nodule cultures

Nodule cultures were induced from shoot tips of aseptically grownEucalyptus botryoides in a vertically rotated incubator. N-(2-chloro-4-pyridyl)-N'-phenylurea (4PU) was a very effective cytokinin for induction of nodules in liquid B5 basal medium supplemented with naphthalene acetic acid and 3% sucrose.The effective concentration of 4PU ranged from 0.2 to 0.5mg/l. The nodule had a very smooth surface and was composed of small meristematic cells outside and large vacuolated cells inside. Shoots were regenerated from these cultures on media supplemented with 6-benzyladenine at 0.2mg/l in place of 4PU. These regenerated shoots were successfully rooted and cultivated in the field. Nodule cultures were maintained for 3-4 years with monthly subcultures without losing proliferation and regeneration abilities. Nodules were also successfully induced from other eucalypts, namelyE. camaldulensis, E. deglupta andE.grandis with slightly modified media. Furthermore, nodule cultures were also induced from shoot tips of field-grown plants inE.camaldulensis. This system is beneficial for both mass propagation of selected elite clones and creation of genetically engineered plants.