Abstract
The experiment describes the establishment of in vitro conditions for seed germination,
micropropagation, callogenesis, organogenesis and acclimatization of Citrullus colocynthis (L.) Schrad,
of family Cucurbitaceae.
Methods: In vitro grown seedlings from decontaminated seeds were micropropagated in basal MS medium at
23±2oC temperature and light intensity of 3000 Lux for 16 hours in culture room. In vitro grown nodal explants
were supplemented with BAP (6-Benzylaminopurine) and NAA (Naphthalene acetic acid) with basal MS
(Murashige and Skoog) medium to induce multiple shoots. Indole butyric acid (IBA; 0.1 to 2.0 mg/L) was
supplemented to MS medium to develop roots of micropropagated shoots. Internodes and leaves of
micropropagated shoots used to induce callus in MS medium enriched with varying concentration of 2, 4-
dichlorophenoxyacetic acid (2, 4 D; 0 to 2.0mg/L) and kinetin (KIN; 0 to 1.0mg/L). Shoot initiation from callus
was tested by adding 2, 4-D (0.1 to 2.0 mg/L) and BAP (1.0 to 1.5 mg/L) in basal MS medium. Conditions were
carefully monitored during the experiment. After hardening, the micropropagated plantlets were placed in open
filed environment in pots filled with sand and peat moss (3:1).
Result: Surface sterilized seeds of Citrullus colocynthis (L.) showed 100% germination in regulator free medium.
Significantly mature shoots (75%) from nodal explant recorded in BAP (2.0 mg/L) and NAA (1.0 mg/L)
augmented MS medium. Highest number (90%) of roots per shoot explant were observed in IBA (2.0 mg/L).
Leaf explants showed better response to form callus with a combination of 2, 4-D (1.0 mg/L) and KIN (1.0 mg/L)
and further rise in 2, 4-D concentration caused a sharp decrease in callus formation. Shoot induction from callus
cultures observed in MS medium containing 2, 4-D (2.0 mg/L) and BAP (1.5 mg/L), producing an average of 10
shoots per culture. Plants were effectively transplanted in open environment with survival rate of 85%.
Conclusion: Results indicate the successful establishment of the growth room conditions for in vitro
micropropagation of the endangered medicinal plant, Citrullus colocynthis.
Arneeb Tariq, Humera Afrasiab, Fozia Farhat. (2020-2021) In vitro Micropropagation of Citrullus colocynthis (L.) Schrad: an endangered medicinal plant, Advancements in Life Sciences, Volume-8, Issue-1.
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