Regenerating rots in Arabidopsis shoots

George Karlin-Neumann karlin at CMGM.STANFORD.EDU
Wed Aug 5 14:46:28 EST 1992


    We use the following procedure for rooting shoots regenerated by the
Valvekens procedure (1988; PNAS,85,5536):

1) shoots (both prior to bolting and after bolting) are broken free from callus
with fine forceps, minimizing the inclusion of callus tissue on the end of the

2) shoots are transferred to hormone-containing rooting medium for 2-4 days to
promote root initiaion (1X MS salts, 1% sucrose, 1X B5 vitamins; 12 mg/L IBA
(final conc.); 0.1 mg/L kinetin) -- NO drugs;

3) shoots are then transferred to new dishes containing the same medium, BUT

Roots begin to evidence within 1-2 weeks after transfer to hormone-free medium,
in at least 50% of the shoots, and are usually transferred to soil about 1 week
after roots first apparent.  This works well with various ecotypes including
Columbia, WS and RLD. (Note: Beware that Carbenicillin -- at least the batch we
have on hand from Sigma -- strongly retards root growth at concentrations of
250ug/ml and above.  If you want to include an antibiotic, timentin seems to be
inocuous in this regard).

                                Good luck, George Karlin-Neumann

More information about the Arab-gen mailing list