IJA -2016v6n8 - page 9

International Journal of Aquaculture, 2016, Vol.6, No.8, 1-10
5
Table 4 Effect of culture medium types and condition on
in vitro
shoot proliferation of
L. antipoda
culture initiation after 28 days
Medium type
Medium strength
Mean number of shoots per explant
Mean length of shoots
Semi solid
MS
5.5±0.22
3.5±0.03
Semi solid
1/2MS
4.5±0.47
4.5±0.52
liquid
MS
6.5±0.23
4.22±0.55
liquid
1/2MS
15.3±0.87
4.8±0.41
Although the response of explants in ½ MS basal medium was not at all satisfactory, excellent responses were
obtained in half strength MS medium supplemented with various concentrations of growth regulators (Table 5).
Results revealed that ½ MS medium supplemented with BAP, Kn & NAA showed significant difference in terms
of shoot proliferation effect. The best response of culture initiation was obtained in half strength MS medium
supplemented with 1 mg/l BAP. BAP concentration 0.1-2 mg/l had highest culture establishment capability (40.3 -
85%) and also culture initiation was very fast within 10 days. But in Kinetin and NAA, culture establishment was
below 40% (MS control 35%) and shoot initiation took more than 10 days. Further studies were focused to find
out the most appropriate medium for maximum shoot proliferation. MS media with Cytokinin proved to be most
effective, for multiple shoot proliferation. After 25-30 days of first subculture, established cultures were
transferred to culture jars having respective media combinations. Multiplication of shoot cultures was carried out
by culturing nodal segments/clusters excised from
in vitro
raised plants.
Table 5 Effect of various concentrations of growth regulators on
L. antipoda
L., in BM
1
- BM
5
Media.
Effect of various concentration of growth regulators on shoot proliferation are provided in the Table 6.
Observations were taken for evaluating the growth of explants by taking parameters like inter nodal distance,
average shoot length and number of nodes. The experiment was carried out in seven media having different
concentrations of growth regulators each with 3 replications; only results of best media are given. The media that
showed best results were BM
3
and BM
5
with highest average shoot length of 5.56 and 6.3 respectively with 100%
cluster formation. Sub culturing was carried out after 25-30 days using the same medium combinations as for
initiation and establishment stages. Shoot clusters obtained in each subculture was divided in appox 1 cm
2
size
with approx 4-5 small shoots in each cluster and inoculated in each bottle.
Sl No
Media
Number of
shoots >3cm
Number of
nodes
Average shoot
length
Average number
of roots
Remarks
1
MS(Control)
3.24±0.41
3.13±0.00
2.8±0.65
2.5±0.25
Least response
2
BM1
(MS+0.1mg/l BAP+0.1mg/l
NAA)
5.19±0.33
4.09±0.30
3.9±0.15
2.8±0.54
Rudimentary
response
3
BM2
(MS+0.5mg/l BAP+0.2mg/l
NAA)
8.07±0.25
5.03±0.45
4.31±0.25
5.0±0.23
Moderate
response
4
BM3
(MS+1mg/l BAP+0.2mg/l
NAA)
16.60±0.25
6.5±0.23
5.56±0.65
10.5±0.6
Maximum rate of
proliferation and
flowering
5
BM4
(MS+2mg/l BAP+0.4mg/l
NAA)
10.62±0.32
6.20±0.55
4.12±0.47
12.5±0.5
Maximum
response with
callusing at base
6
BM5
(MS+1mg/l BAP+0.5mg/l
Kn)
12.60±0.14
5.25±0.57
6.30±0.36
-
Moderate shoot
length and
multiplication
7
BM6
(MS+0.5mg/l BAP+0.5mg/l
Kn)
6.60±0.55
4.62±0.33
5.50±0.2
-
Moderate
response with no
roots
1,2,3,4,5,6,7,8 10,11,12,13,14,15,16
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