IN VITRO CULTURE OF BRAHMI | Online Biotech Notes

 

Online Biotech Notes

IN VITRO CULTURE OF BRAHMI : A CASE STUDY

IN VITRO CULTURE OF BRAHMI

 

1. Selection of Explant:
• For in vitro culture of Brahmi, we selected a healthy and disease free mother plant from the field or green house.  Brahmi is micro propagated through inter node and nodal explants.
2. Excision of Explant:
• Shoot tip and nodal explants were excised by a sterile scalpel to get a 2cm. Explant containing meristematic tissue (buds).
3. Sterilization of Explants:
• The explants were sterilized by following ways as: 
• The explants were washed thoroughly with tween- 20 for 10 minutes with constant shaking & then washing under running tab water. Then explants were treated with fungicide Bavistin (0.2%) and Bacterialcide Sterptocycline (0.2%) for 90 minutes with regular shaking.  Rinsed explants with distilled water. The Subsequent steps are done in front of a laminar air flow or the pressurized inoculation chamber.
4. Preparation of Media: 
• Media prepared was basal MS media with combination of hormones.  Hormone   BAP (2 mg/l) was added in medium. 30g/ltr. Sucrose was added.  PH was set to 5.8.  Then boiled media to 60^°C and added 8gm/l Agar.  Then media was sterilized in autoclave for 15 minutes at 121°C and 15 lb/inch² and then stored. 
5. Inoculation of Explant:            
• Inoculation was done in laminar air flow.  First UV was switched on for 10 minutes.  Then UV light was switched off.  The surface of laminar cabinet was disinfected by wiping with absolute alcohol. Explants were brought to LAF & treated with 70% alcohol followed by rinsing with sterilized double distilled water for 4-5 times, the explants were then treated with 0.1% HgCl₂ for 5minutes followed by rinsing with double distilled water for 4 to 5 times. The explants were then inoculated onMS medium supplemented with various growth regulators.  The bottles were Marked and kept in to growth rooms.

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