International Journal of Aquaculture, 2016, Vol.6, No.8, 1-10
3
for 16/8 hrs of day/night break under the cool white fluorescent light of average 2500 lux (cool white fluorescent
tube light 40 W GE).
2.5 Axillary shoot proliferation
Multiple shoots/cluster were transferred from the culture bottle to a sterile glass plate; brown leaves were removed
from the primary shoots and sectioned into one node piece after removing the leaves. These nodal segments were
transferred to the multiplication media (Table 2. These culture bottles were then incubated in the growth room.
These steps were repeated every 25-30 days for the next sub-culturing.
Table 2 Description of various media for the shoot multiplication of
L. antipoda
(L.)
Medium Code
MS*
Medium Details
MS basal as control
BM1
MS+0.1mg/l BAP+0.1mg/l IAA
BM2
MS+0.5mg/l BAP+0.5mg/l IAA
BM3
MS+1mg/l BAP+0.2mg/l IAA
BM4
MS+2mg/l BAP+0.4mg/l IAA
BM5
MS+0.5mg/l BAP+0.5mg/l Kn
BM6
MS+1.0mg/l BAP+1.0mg/l Kn
2.6 Transplantation and acclimatization of the plantlets
After 10 – 14 days of culture on rooting media, the rooted plantlets were transplanted to pots or trays for
hardening prior to their final transfer to aquarium. Rooted plantlets were taken out of the culture bottles with the
help of forceps and washed thoroughly with water to remove any remaining of the medium. Bavistin (0.1%)
treatment was given to the plants in order to protect them from fungal attack in the near future. After this, the
plants were carefully planted in poly bags containing different ratio of Neopete (Sterlings Farm Research Services
Pvt. Ltd.) & soil mixtures in 1:1 ratio. After planting, plantlets are thoroughly watered and kept in poly house
under humidity range of approximately 80%. These plantlets were sprinkled with water time to time as per the
requirement and after two weeks and transferred to shade house having humidity range of approximately 60%.
The plantlets are then transferred to open area after 9-10 days and kept there for ten days prior transferring them to
the aquarium.
3 Discussion
3.1 Surface sterilization of explant
Sodium hypochlorite solution (NaOCl, 4% available chlorine, Fisher Scientific Prod. No. 27908) was used for
surface sterilization of explant. After treatment with sodium hypochlorite over 40% of the cultures were
contaminated by fungi and or bacteria. Response of
L. antipoda
(L.)
explants to surface sterilisation with various
concentration of NaOCl is provided in the Table 2. From the table, it was clear that 10% NaOCl for 10 minutes
was the best surface sterilization procedure for
L. antipoda
(L.)
explants. In this, 60.2% explants survived and
39.8% were contaminated. Use of 10% NaOCl for 20 min caused the death of explants (80%). The survival rate
was only 10.4% but had lower contamination rate 9.6%. Hence, 60.2% survival rate on treated with 10% NaOCl
for 10 min, the contamination rate was 35.5%, to avoid this systemic contamination bavistin was effectively used.
By pre-treatment of the explants using 0.15% of bavistin for 3 minutes ceased the rate of contamination to 15.5%
and sprouting percentage of shoots raised to 84.5%. But, when bavistin with 0.3% concentration was used,
contamination rate was reduced to 6% while rate of sprouting also ceased to 48%. The variable response of
Lindernia
sp. explants to bavistin
treatment is provided in Table 3. It is proved that various concentration of
bavistin was tried and 0.15% was optimum for initiating a better response in
L. antipoda
(L.)
