Vegetative propagation of elite Eucalyptus clones as food source for honeybees (Apis mellifera); adventitious roots versus callus formation: supplementary material

Summer and autumn in Israel are highly arid with not enough plants in bloom offering nectar and pollen to support the local apiary. This leads to decline in colony health and honey production. To increase food sources for honeybees, we initiated a project to clone elite Eucalyptus trees exhibiting constant and rich blooming from late summer to early winter. We induced adventitious roots from cuttings of two mature Eucalyptus trees of which nectar production and honeybees' attraction was measured: Eucalyptus brachyphylla and Eucalyptus x trabutii. During the rooting process, a high frequency of cylindrical callus formation instead of roots was obtained. To shed light on the inner anatomy of the callus chunks, we compared their cell organization and cell-wall composition to those of roots. Whereas in the root, cells were organized in circumferential symmetry, no symmetry was found in the callus. Instead, a more chaotic accumulation of meristematic-like cells with sporadic clusters of tracheary elements laid in different directions were observed. The outer cell layer of the callus often included swollen cells with thin cell walls. Most callus cells stained more strongly for cellulose and lignin than cells in the root meristem. In addition, specific antibodies to methylesterified and de-methylesterified pectin showed differential staining of callus vs. root cells indicating cell wall differences. Strikingly, roots were seen to differentiate from the chaotic cell organization of the callus, albeit at low rates. Further investigation of the cellular and molecular mechanisms underlying callus formation, are required.