"Tissue Culture Trials with Jatropha"

 

Seed germination

In vivo germination

  • Seeds were washed with running tap water for 6 hrs/day for two days

  • The washed seeds were cultured in media composed of perlite: peat moss: sand 1:1:1

 

 

 

In vitro germination

  • The seeds were soaked in 20% HCl for 15 min, and then

  • Washed three times with sterilized distilled water

  • Cultured in jars contained cotton saturated with MS medium (growth regulators free)

  • Incubated in the growth room

 

 

Micro-propagation

Establishment stage

Explants sterilization

  • Nodal explants were washed in soap water using septol soap for 30 min. and subjected to continuous flow of tap water for about 40 min. There after, under aseptic condition using laminar airflow cabinet, they dipped in 70 % ethanol for 60 sec.

  • The explants were sterilized using commercial bleach Clorox at  20 %  (v/v) solution (NaOCl 5.25% of free chloride)  for 15 min. with a few drops of tween-20

  • Then they were rinsed three times in sterile distilled water.

 

Direct organogenesis through axillary buds

  • Nodal explants were cultured on MS media supplemented with Benzyl Amino Purine (BAP)  and Adenine sulfate.

  • Then incubated in the growth room at 26+2 °C for a period at specific light intensity and light/dark periods of 16/8 hours.

 

 

Bud growth after establishment

Multiplication stage

·         The growing shoots were sub-cultured on MS media supplemented with different doses of different plant growth regulators

 

 

 

Indirect regeneration throgh callus induction

  • Leaf explants was washed in soap water using septol soap for 30 min. and subjected to continuous flow of tap water for about 40 min. There after, under aseptic condition using laminar airflow cabinet, they dipped in 70 % ethanol for 60 sec.

  • The explants were sterilized using commercial bleach Clorox at 10 %  (v/v) solution (NaOCl 5.25% of free chloride)  for 20 min. with a few drops of tween-20/

  • Then they were rinsed three times in sterile distilled water and cultured on MS medium supplemented with different concentrations of growth regulators.

 

 

Callus formation from leaf explants