need help with root culture

[DEPARTAMENTO DE MICROBIOLOGIA]

Dear Tanya,

I have no worked with poplar but I have established root
cultures of some crops like tomato and alfalfa. Maybe you can
try the same protocol:

- Surface sterilise the seeds (1 min ethanol 95% + 20 min in
comercial bleach + three baths of 5 min. each in sterile
distilled water)

- Transfer the axenic seeds to flaks previously autoclaved
containing humid cotton.

- cut 1 cm of the emerging root (including the root tip) and
transfer it to 250 ml Erlenmeyer flasks containing 50 ml of
autoclaved MS medium + sucrose 3%. Incubate in an orbital
shaker 130 rpm aprox. 25† C.

- Usually the roots growth an multiply producing lateral roots
that can be cuted an subcultured in fresh media.


																		Best wishes,


																	Xabier Barandiaran