Abstract:
Nymphaea is commercially important aquatic plants propagated through
cross pollinated seeds cause trait variations in the offsprings, the
morphological features of flowers are different from one plant to another and
cause major problem by commercial producers and vendors. Therefore,
multiplication through micropropagation is a viable option. Hence, this
research focused on establishing a robust in vitro protocol for Nymphaea leaf
culture. The young leaves of Nymphea were excised from a single mother
plant and cut into (5mm x5mm) pieces were used as explants. Then five
sterilization methods (Factor 1) viz: 0.2% of HgCl2 (T1), 0.1% of HgCl2 (T2),
5% NaOCl (T3), 10% NaOCl with Tween20 (T4), 10% NaOCl (T5) were
employed. Then two types of media (Factor 2), namely 1/2MS (M1),
comprising 50 ml of A stock, 2.5 ml of B stock, 5 ml of C stock, along with
glycine, pyridoxine, nicotinic acid, thiamine, BAP, 2.4-D, Myo-inositol, and
sugar, consistent and favorable results were observed across various
sterilization techniques. Conversely, media 2 (M2), with a similar
composition but supplemented with BA, NAA, and kinetin were employed.
The data collected over the course of 12 weeks, the results indicate that T4
consistently exhibits the lowest contamination percentage of leaf across both
media formulations, while T3 consistently demonstrates higher
contamination rates. Furthermore, media 1 consistently yields superior
results in terms of tissue culture initiation compared to media 2. By
identifying optimal sterilization techniques and media formulations, this
study provides valuable insights for enhancing mass propagation.
