Protocols for Micropropagation of Woody Trees and Fruits

238
programmes is somatic embryogenesis (Conde et al., 2004). The presented protocol
describes an efficient method for in vitro micropropagation of wych elm (Ulmus
glabra Huds.), smooth-leaved elm (U. minor Gled.) and European white elm (U.
laevis Pall.) by the axillary bud culture method (see Figure 1).
Figure 1. Stand of Ulmus glabra donor plus trees in Ore Mountains.
2.1. Explant Preparation
J. MALÀ ET AL.
2. EXPERIMENTAL PROTOCOL
2.1.1. Growing Condition of Mother Plants
For the establishment of primary elm cultures the plant material from 40 to 80 years
old donor plus trees of Ulmus glabra, U. laevis, and U. minor (15 clones from each
elm species) has been used. The origins of the different elm species are summarized
in Table 1.
2.1.2. Explant Excision and Sterilization
Dormant axillary buds collected in February were used for tissue culture establishment.
Thirty buds have been taken from each clone for primary cultivation. The
buds were first rinsed under tap water to remove surface dirt, and sterilized by
immersion in 0.1% NaClO3 for 10 min followed by immersion in 0.01% HgCl2 for
15 min. The buds were further rinsed three times with autoclaved distilled water for
15 min. Finally, the apical meristems were excised from buds by means of sterile
incers and scalpel.