A.thaliana plants dying after transformation procedure

Estelle Hrabak emhrabak at cisunix.unh.edu
Wed Aug 2 08:37:58 EST 2000


>hi all,
>i am trying to transform arabidopsis plants with my promoter constructs
>in Agrobacterium tumefaciens GV3101 using the protocol on andrew bent's
>website. The first time i used the recommended 0.05% Silwet L-77 and my
>plants all died. The second time my plants died inspite of lowering the
>conc. of the silwet to 0.03%. Any suggestions to what i can change in
>the protocol from people with similar experience.
>zarir
>

One thing that might help with plant viability is to rinse off the plants
and gently leach through the soil in the pot with running water the morning
after dipping.   (That would be after step 7 on Andrew Bent's web
protocol).   I think that what happens sometimes is that if too much
residual sucrose from the dipping solution is left on the leaves, or
especially in the soil, it can provide a food source for fungi or bacteria,
with negative effects on plant growth.  I think that I've seen less
problems when the soil is not extremely wet on the dipping day or when one
is very careful to let only the aerial parts of the plant come in contact
with the dipping solution and not the soil.

Estelle


Estelle M. Hrabak
Assistant Professor
Dept. of Plant Biology
University of New Hampshire
46 College Rd.
Durham, NH  03824

Ph.  (603) 862-0716
FAX (603) 862-3784



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