Basic HTML Version
Tree Genetics and Molecular Breeding 2014, Vol.4, No.1, 1
-
6
http://tgmb.sophiapublisher.com
3
1.2
In vitro
rooting in regenerated shoots
In vitro
rooting was dependent on the strength of the
medium and the auxin concentration as evident from
Table 3. Among the various strength of media
evaluated, IBA, IAA and BAP levels; ½ MS+IBA
8.88+IAA 1.14+BAP 0.89 µM+Activated charcoal
500 mg/L produced 88.16% rooting with heighest
number of roots (4.33 roots/explant) over other treat-
ments. In MS+IBA 13.33+IAA 1.14+BAP 0.89 µM,
regenerated shoots produced longer roots (5.26 cm).
Table 3 Effect of different concentration of auxins and cytokinins on
in vitro
rooting
Treatment (µM)
Rooted explants
(% )
No. of roots/
explant
Root length/
explant (cm)
MS
IBA 0.0+ IAA 1.14+BAP0.89+Activated
charcoal 500mg/l
0.00
0.00
0.00
IBA 9.85+ IAA 1.14+BAP0.89+ Activated
charcoal 500mg/l
62.56
1.33
3.26
IBA
14.77+
IAA
1.14+BAP0.89+Activated
charcoal
500mg/l
52.40
2.33
5.26
IBA
19.70+
IAA
1.14+BAP0.89+Activated
charcoal
500mg/l
44.83
1.66
3.56
SE(m)±
0.32
0.29
0.11
CD (0.05)
1.06
0.95
0.38
½ MS
IBA 0.0+ IAA 1.14+BAP0.89+Activated
charcoal 500mg/l
0.00
0.00
0.00
IBA 9.85+ IAA 1.14+BAP0.89+Activated
charcoal 500mg/l
88.16
4.33
4.26
IBA
14.77+
IAA
1.14+BAP0.89+Activated
charcoal
500mg/l
75.03
2.66
3.46
IBA 19.70+ IAA 1.14+BAP0.89
66.16
2.33
3.16
SE(m)±
0.45
0.29
0.12
CD (0.05)
1.50
0.95
0.41
Note: Values in these columns indicates average values of 3 replicates
1.3 Hardening of
in vitro
plantlets
Acclimatization of the rooted plantlets was tested in
various carrier substrates (Figure 1). A combination of
Soil+Sand+Peat moss (1:1:2) supplemented with ½
MS nutrient solution was identified to be ideal
substrate for acclimatization, as this treatment resulted
in maximum plant survival rate (88.33%), longer
plants (6.56 cm) with healthy and green canopy of
leaves (8.33 leaves/plant). Although, longer roots
(5.96 cm) were found in coconut husk carrier this
treatment did not show efficient survival rates.
Totally 46 plantlets survived in the greenhouse after
one year.
2 Discussion
2.1 Shoot bud induction and multiplication
Micropropagation of mature trees using vegetative
parts as explants has been a difficult task due to
various factors, like juvenility vs. maturity, inherent
slow growing habit, exogenous and endogenous
Figure 1 Effect of carrier substrates on acclimatization of
micropropagated plantlets
infection, presence of phenolic compounds, long
complex life cycles, great genetic variations, etc.
(Bonga and Durzan, 1986; Bajaj, 199; Ali et al., 2003;
Liu and Yang, 2011).
In vitro
propagation of plant
species is influenced by factors such as genotype, age