Standardization of Protocol for the Surface Sterilization and Callus Induction of Saussurea lappa: An Endangered Medicinal plant
By: Zaib Un Nisa, Samin Jan, Safdar Hussain Shah
Key Words: Saussurea lappa, Explants Sterilization, Callus Induction.
Int. J. Biosci. 9(1), 324-330, July 2016.
Saussurea lappa is an endangered medicinal plant, used against lot of diseases like fever, headache, pain, arthritis, ulcers, cough, flatulence, asthma, colic, diarrhea, dyspepsia, vomiting, tenesmus, and dysentery. The plant is endangered due to over exploitation for commercial and medicinal purposes. The study was carried out to develop a protocol for the surface sterilization and callus induction of the seeds of S. lappa. Seeds of the plant were used as explants for the sterilization and callus induction. The explants were surface sterilized with 5% solution of tween-20 for 20 minutes followed by three different concentrations (0.05%, 0.1% and 0.2%) of mercuric chloride for 8minutes in three different experiments and rinsed 5 times thoroughly with autoclaved distilled water. Results were shown in percentage. The concentration of 0.2% solution of mercuric chloride showed best sterilization (96%) compared to 0.1% (48%) and 0.05% (0.2%) solution of mercuric chloride. Solid Murashige and Skoog (MS) medium fortified with 1mgL-1Dichlorophenoxyacetic acid (2-4 D) alone and in combination with 0.25 mgL-1 kinetin (Kn) were used for the callus induction. Relative Growth Rate (RGR) of the callus was determined after 15 days of inoculation and found that the combination of 2-4 D and Kn produced maximum (0.141week-1) callus compared to 2-4D (0.083 week-1) used alone. The results showed that 0.2% HgCl2 solution is the best seed sterilizing agent while 2-4 D (1mgL-1) and Kn (0.25mgL-1) are the best combination for the callus induction and fresh weight bio mass.