Shoot Induction and Somatic Embryo Development
Lab #1

Murashige and Skoog (MS) salts
Gamborg B5 vitamins
3% sucrose

and either
1) no addition “OMS”
2) 170 mg/l NaH₂PO₄×H₂O, 80 mg/l adenine sulfate, 0.5 mg/l NAA, 2 mg/l kinetin “AV”

 FOR 1.0 LITER
10 ml    MS FeEDTA  add first
10 ml    MS Sulfates
10 ml    Halides
10 ml    P, B, Mo
10 ml    B5 Vitamins
1.65 g  NH4NO3
1.9 g    KNO3
30 g sucrose

volume up to 1000 ml
split into 2 x 500 ml

 add 1.0 g Gelrite

 autoclave, pour into plates

For the AV medium, the explants are the leaf blade and petiole of African violet and streptocarpus. Leaf tissue will be surface sterilized for 15 minutes with 15% bleach containing 50 µl Tween-20 per 100 ml. Leaves will be washed 4 times with sterile distilled water. The effects of explant orientation and size can be evaluated. Shoots will be visible within 2 weeks.

For OMS, proliferative embryogenic tissue of soybean will be placed on this medium and embryo development will be observed over the next 4 weeks.

RESULTS

 
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