Liquid Culture Systems for in vitro Plant Propagation

Application of temporary immersion in Phalaenopsis 233
by Escalona et al. (1999). The immersion of shoots was adjusted to eight
times per day for ten minutes each. Spent medium blackened by phenolic
compounds (Figure 1 A) was substituted in intervals of two or four weeks.
The experiments using the twin vessels were repeated three times each. The
multiplication rate and increase of fresh weight were determined twelve
weeks after inoculation. An overview of the experimental design is given in
figure 1 B.
Control cultures were grown on agar gelled medium (Tanaka and
Sakanishi, 1985) supplemented with 10 mg l -1 BAP and 10 % (v/v)
homogenised bananas.
2.2 Rooting
In the first series of experiments shoots of 4 – 7 cm size from TIS
cultures were exposed to TDZ-free medium supplemented with 0.5 and 1.0
mg l -1 IAA or NAA. In the main experiments 1.0 mg l -1 IAA-containing
medium was used. Each vessel was inoculated with 72 – 74 shoots of 70 g
total fresh weight. The immersion frequency was adjusted to two, four or six
times per day for ten minutes periods. The experiments using the twin
vessels were repeated three times each. The percentage of rooted plants and
the increase in fresh weight were determined after eight weeks.
Figure 1: (A) Twin glass vessels (5 l) inoculated with 20 g shoot fresh weight of
Phalaenopsis cv. Jaunina. After two weeks the medium was blackened by phenolic
compounds. (B) Experimental design of a series of twin glass vessels after 12-week culture.