International Journal of Aquaculture, 2016, Vol.6, No.8, 1-10
6
Table 6 The effect BAP and IAA supplemented into MS medium on the shoot proliferation of
L. antipoda
L after 4 weeks of culture
Growth regulators (mg/L)
Number of shoots
(explant
-1
)
Shoot height (cm)
Number of roots
(explants
-1
)
BAP
NAA
0.0
0.0
1.8±0.42
2.4±0.22
0.7±0.4
0.1
0.0
4.0±0.22
3.3±0.27
6.0±0.19
0.5
0.0
15.5±0.56
4.4±0.55
18.5±0.32
1.0
0.0
24.4±1.92
5.5±0.36
30.7±0.45
1.5
0.0
13.0±1.25
4.36±0.58
13.0±0.26
2.0
0.0
14.5±0.35
2.8±0.25
20.5±0.36
2.5
0.0
6.75±1.65
1.52±0.65
0.88±0.12
0.0
0.5
1.0±0.54
2.5±0.27
0.6±0.35
0.1
0.5
3.65±0.26
3.35±0.56
9.0±0.78
0.5
0.5
14.1±0.32
4.2±0.65
16.5±0.33
1.0
0.5
19.5±1.52
4.8±0.65
20.2±0.23
1.5
0.5
11.5±0.67
4.32±0.12
1.23±0.65
2.0
0.5
13.7±0.26
2.7±0.85
18.4±0.56
2.5
0.5
7.2±1.23
1.31±0.26
0.45±00
It was observed that the highest number of shoots (>3 cm) originated from the basal node and average number of
shoots formed in case of BM
3
media (MS+1mg/l BAP+0.2mg/l NAA) is 16.6 ; 5.19 in BM
1
(MS+0.1mg/l
BAP+0.1mg/l NAA); 10.62 in BM
4
(MS+2mg/l BAP+0.4mg/l NAA) and lowest number of shoot regenerated in
case of MS(Control) was 3.24. In term of number of nodes/explants, same order followed i.e. (BM
3
MS+1mg/l
BAP+0.2mg/l NAA) having 6.5, BM
1
(MS+0.1mg/l BAP+0.1mg/l NAA) having 4.09, 6.20 in BM
4
(MS+2mg/l
BAP+0.4mg/l NAA) and in case of MS (Control) was 3.13.
3.2
In vitro
flowering
In vitro
conditions including media components, concentration and ratio of plant growth regulators and culture
conditions affected
in vitro
flowering. Flower initiated in MS media supplemented with 1 mg/l BAP & 0.2 mg/l
NAA (BM
3
) produces multiple shoots with flowering after 4–6 weeks of cultures (Figure 4). The production of
flowering shoots continued for many subcultures spanning a period one year.
In the present study, incubation of flowering cultures on BAP supplemented with NAA in the medium was
necessary for flowering after 6 weeks. Photoperiod was found to be important for
in vitro
flowering. Maximum
in
vitro
flowers were found at 16 hrs ± 2 light periods; it was observed that plants incubated under 12 hrs or shorter
photoperiods (8 hrs) were negatively affected for floral bud development. Optimum temperature for efficient
in
vitro
flowering was 25°C ±2°C.
3.3
In vitro
rooting and hardening
Rooting was also observed along the medium. Highest number of root observed on the BM
4
media is 12.5 due to
0.4 mg/l NAA. Small amount of rooting was observed on BM
1
, BM
2
& BM
3
is respectively. i.e., NAA along with
BAP also stimulates rooting in the medium.
It was observed that during each passage, the number of leaves/shoots has increased substantially along with the
height of shoots. The leaf size was approximately 0.5-0.8 cm. It was observed that morphogenic responses
exhibited in the form of shoots or roots are correlative to a specific auxin/cytokinin ratio. In comparison to MS
basal media, for BM
3
, there is a significant increase in number of shoots (5.28 times) and average shoot length
(3.14 times).
