Protocols for Micropropagation of Woody Trees and Fruits

398
Shoot Number Shoot Length (mm)
30.0
25.0
20.0
15.0
10.0
6.0
5.0
4.0
3.0
2.0
K. PRUSKI
15.0
0 5 10 15 20 0 5 10 15 20
SN - Mongolian cherry
SL - Mongolian cherry
1.0
0 5 10 15 20 0 5 10 15 20
BA Concentration BA Concentration
Figure 3. BA concentration effect on shoot number and shoot length in Mongolian Cheery
and Nanking Cherry. Actual values (solid circles) and the fitted (solid lines) (Pruski et al.,
2005).
can also be used successfully for shoot proliferation of Mongolian and Nanking
cherry cultures, however at a very low level. As indicated in Table 3, compared to
controls (no TDZ), the number of shoots is almost doubled on media containing 0.45
µM TDZ in both species after 4 weeks in culture. Higher TDZ concentrations induce
much more shoots, but their length decreases drastically. Besides these short shoots,
a mass of stunted dark green dwarf shoots are produced by cultures. Such responses
to TDZ have been reported by a number of researchers in a variety of woody plant
species (Bates et al., 1992; Sarwar and Skirvin, 1997; Thengane et al., 2001). Based
on the observations from our studies, a TDZ concentration of 0.5 µM is optimal for
production of good quality Mongolian and Nanking cherry shoots. TDZ has been
reported to be a potent growth regulator of in vitro plant morphogenesis (Mante
et al., 1989; Bates et al., 1992; Murthy et al., 1998).
Procedure for multiplication of shoots:
1. Transfer the established rosette cultures from initiation media in test tubes
to proliferation media in Magenta GA7 vessels: MSMO supplemented with
18 µM of BA for Mongolian cherry and with 15 µM of BA for Nanking
cherry, or with TDZ at 0.5 µM for both species.
2. Place five rosettes per vessel.
3. Incubate cultures for 4 weeks in a growth-room under conditions described
in section 2.2.
4. Repeat the procedure until the desired number of shoots/rosettes for rooting
is achieved.
5. Place the proliferating cultures on hormone-free media 4 weeks prior to the
rooting stage.
27.0
24.0
21.0
18.0
4.2
3.4
2.6
1.8
SN - Nanking cherry
SL - Nanking cherry