Date:

   Name:                                                            

   Topic:                          Herb Micropropagation

                         Induction of the Culture

   

   Material: Leaves of African violet (Saintpaulia ionantha Wendl.) and Cape primrose
   (Streptocarpus x hybridus) (Family Gesneriaceae)

   Medium: F1 = M-S basal salts, B5 vitamins, BA 1mg.l^-1, NAA 1mg.l^-1, sucrose 20 g.l^-1, 0.65%
   agar, pH 5.5. Recalculate concentrations of plant growth regulators and sucrose to molar
   concentration (= SI units).

   

   Procedure:

   1.    Remove fully expanded leaves from a stock plant and wash them with distilled water.

   2.    Disinfections of the leaves in 70% ethanol for 30 sec to1 min.

   3.    Disinfections in aqueous solution of 15% commercial bleach SAVO (v/v) by gentle
   agitation for 15 min.

   4.    Rinse 3 times in sterile distilled water for 3 min.

   5.    Transfer disinfected leaves from the bottle on a sterile Petri dish in a flow hood.

   6.    Excise 1cm^2 segments of the leaf.

   7.    Place the segments with the adaxial surface of the leaves (upper layer of epidermis of
   the leaf) in contact with the agar medium F1.

   8.    Record the number of segments within a tissue culture jar and the number of the jars.

   9.    Culture under the light (cool white fluorescent light, photoperiod 16/8 hours, PAR 30
   μmol.m^-2.sec^-1) at 22-oC for 4 weeks.

   

   Evaluation:

   Control contamination of the explants in the following period of cultivation. The contaminated
   cultures should be discarded.

   Evaluate frequency of segments with organogenic shoot bud formation (direct organogenesis) and
   the number of segments with proliferating callus stage (indirect organogenesis). Record the
   multiplication rate (total number of shoots per explant).

   

   http://www.africanviolet.org.au/streptocarpus.htm

   http://www.robsviolet.com/streptocarpus1.htm

   http://www.streptocarpussociety.org.uk/

   http://en.wikipedia.org/wiki/Streptocarpus

   http://www.ext.vt.edu/departments/envirohort/factsheets/pottedplants/strept.html