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Bioscience Methods 2012, Vol.3, No.3, 21
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Table 2 Effects of mediums with different concentrations of sucrose for callus induction
Treatment
Medium formula
No. of explants
Growth status of calli
Rate of callus
induction (%)
Day of callus
initiation (d)
Status of calli
1
MS+sucrose 30 g/+KT 0.5 mg/L +
6-BA 0.5 mg/L+NAA 0.5 mg/L
100
78
9
Pale-yellow, dense
2
MS+sucrose 30 g/L+6-BA 0.5 mg/L
+NAA 0.5 mg/L
100
79
9
Pale-yellow, dense
3
MS+sucrose 50 g/L+KT 0.5 mg/L
+6-BA 0.5 mg/L+NAA 0.5 mg/L
100
82
6
Yellow green, loose
4
MS+sucrose 50 g/L+6-BA 0.5 mg/L
+NAA 0.5 mg/L
100
88
6
Yellow green, loose
Figure 1 Callus induced from
R. officinalis
L. leaf explants
Note: A: Calli on the medium with treatment 2 after 20 days; B:
Calli on the medium with the treatment 4 after 20 days
in medium was able to dedifferentiate to adventitious
buds. The results demonstrated that the MS medium
with 6
-
BA 1.5 mg/L, KT 0.5 mg/L and NAA 0.5 mg/L
was proved to be the optimal medium with 50 %
regeneration rate, and the number of adventitious buds
ranks first place (Table 3).
1.4 Propagating of adventitious shoots
In order to reduce the teratogenic effect on the tube
seedlings by the plant hormones accumulation, it was
beneficial to reduce the cytokinins and auxin
concentration (Table 4). Results indicated that MS
medium with 6
-
BA 0.8 mg/L and NAA 0.5 mg/L was
proved to be the best medium and the efficiency can
reach 356% (Figure 2).
1.5 Rooting induction for
R. officinalis
L. shoots
When obtaining enough adventitious buds, it was suitable
to reduce the plant growth regulators concentration,
Table 3 Regeneration of
Rosmarinus officinalis
L. callus
Treat-ment
Medium formula
No. of the explants
Observed result
Rate of budding (%)
The total No. of budding
1
MS+KT0.5 mg/L+6
-
BA 1.0 mg/L+
NAA 0.1 mg/L
40
10
4
2
MS+KT 0.5 mg/L+6
-
BA 1.5 mg/L+
NAA 0.1 mg/L
40
20
15
3
MS+KT 0.5 mg/L+6
-
BA 2.0 mg/L+
NAA 0.1 mg/L
40
15
9
4
MS+KT 0.5 mg/L+6
-
BA 1.0 mg/L+
NAA 0.3 mg/L
40
20
23
5
MS+KT0.5 mg/L+6
-
BA 1.5 mg/L+
NAA 0.3 mg/L
40
35
58
6
MS+KT 0.5 mg/L+6
-
BA 2.0 mg/L+
NAA 0.3 mg/L
40
25
32
7
MS+KT 0.5 mg/L+6
-
BA 1.0 mg/L+
NAA 0.5 mg/L
40
25
26
8
MS+KT 0.5 mg/L+6
-
BA 1.5 mg/L+
NAA 0.5 mg/L
40
50
72
9
MS+KT 0.5 mg/L+6
-
BA 2.0 mg/L+
NAA 0.5 mg/L
40
35
50