Growth Regulators

auxins and cytokinins

gibberellic acid
abscissic acid
 pH of media
usually 5.0-5.7
 Media
must be sterile
autoclave at 250 F at 15 psi
for 15 minutes
 T.C. Stages
Explanting- Stage I
get plant material in sterile
culture so it survives
provide with nutritional and
light needs for growth
 Stage II
rapid multiplication
stabilized culture
goal for a commercial lab
difficult and time consuming
to maintain
 Stage II
occurs in different pathways
in different plants
 Rooting - Stage III
may occur in Stage II
usually induced by changes
in hormonal environment
lower cytokinin concentration
and increase auxin
 Rooting
may skip stage III and root
in a greenhouse
 Stage IV
transplantation and
aftercare
usually done in greenhouse
keep RH high (relative
humidity)
 Stage IV
gradually increase light
intensity and lower RH after
rooting occurs
allows plants to harden and
helps plants form cuticle
 Cuticle
waxy substance promotes
development of stomates
plants in T.C. don’t have
cuticle
 Explant
portion of plant removed and
used for T.C.
Important features
size
source - some tissues are
better than others
 Explant
species dependent
physiological age - young
portions of plant are most
successful
 Explant
degree of contamination
external infestation - soak
plant in sodium hypochlorite
solution
 Explant
internal infection - isolate
cell that is not infected
roots - especially difficult
because of soil contact
 Explant
herbaceous plants
soft stem
easier to culture than
woody plants
 Patterns of
multiplication
stage II - light 100-300 foot
candles
callus - shoots - roots
stage III - rooting - light
intensity 1000-3000 foot
candles
 Genetic
transformation
permanent incorporation of
new or foreigh DNA into
genome of cell
 Transformation
methods
protoplast fusion
cell wall is enzymatically
removed from cell
 Protoplasts
naked plant cells
from 2 different plants can
be mixed together and
forced to fuse