Regenerating rots in Arabidopsis shoots

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


Jim,

    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
shoot;

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
LACKING HORMONES;

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
(Stanford)
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