Saturday, 14 March 2015

Bacopa Monnieri (Brahmi) Plant Tissue culture



"Plant Tissue culture 
Of 
Bacopa monnieri"
(Brahmi)

Steps-1- Treatment & Washing of Explants

  • Select a healthy plant from field.
  • And collect the shoot parts.
  • Wash the shoots thoroughly with running tap water.
  • Again wash with few drops of tween-20, thoroughly for removal of fungal or bacterial spores or 1% extran treatment for 10 to 15 min.
  • Washed with DDW for 3 to 4 times.
  • If treated with 1 % extran then followed by 1% Bavistin treatment for 20 to 30 minutes and again washed with DDW 5 times.
  • Transfer the Explant in Pretreated Laminar flow compartment.

Steps-2- Laminar Air flow Treatment

·         1st UV treatment of LAF- Vacant LAF properly rinsed by alcohol and kept 40- 60 min for U.V. Treatment.
·         2nd U.V. Treatment- After 1st U.V. treatment all the required equipment like scalpels, force, sissor, SDDW, petridish and media was surface rinse by alkohal and kept in to LAF for min.

Step-3- surface sterilization & Trimming of Ex-plant under LAF

  • Surface sterilized with 0.1% HgCl2 for 5 min followed by 4-5 times rinsing with double distilled water in Laminar air flow.
  • Ex-plant was trimmed with the help of scalpel and forceps to remove the exposed part of the explants.

Step-4- Inoculation

·         Transfer the material over the flame with the help of forceps in MS media test tube or bottle.
·         Cover it with cap.
·         Seal it with tape and label the culture vessels with the name of the explants and dote of media preparation and date of inoculation.

Step-5- Transfer in culture Room and culture Room condition

·         Material thus inoculated is kept in the culture room (culture room at 25 ± 2  ͦC under 16 hrs light(1000 lux of light intensity and 8 hrs dark condition.

Step -6- sub culturing

·         After establishment of culture in the culture tubes they are aseptically transferred or inoculated in wide mouth bottles for further development.

Step-7- Hardening

·         After proper development of plantlets, they are transferred to Mist. Chamber/ green house/Shade house for hardening or acclimatization.

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