T-DNA transformed pools - INFORMATION

Randall L Scholl rscholl at magnus.acs.ohio-state.edu
Wed Jul 8 10:16:34 EST 1992



     As previously annouced on this network, the pooled tranformed
lines and individual visible mutants, generated by Agrobacterium
seed transformation in our laboratory, are being distributed
through the stock centers. Following is a short description of the
pooling procedure.

     To pool the transformed lines, approximately 20 seeds were
taken from each of 20 T3 families (F2 equivalent).  The 400 seeds
representing these 20 transformants were grown in one 37 x 51 cm
flat (these will be referred to as a subpool).  These plants were
grown under short day conditions such that maximum seed yield
would be achieved.  5,260 transformants, including the 460
transformants generated at Zoecon in 1986-87, and 4,800 generated
at Arizona in 1991, were grown in this manner.  We generally
harvested between 600,000-800,000 seeds from each of these flats
(3,000-4,000 seeds/plant).  However for 23 flats for which the
seed yield was low or for which other difficulties arose, we were
not able to distribute seeds.  To simplify the pooling process for
the formation of the  size-100 pools, Randy Scholl and I decided
to use the Zoecon subpools to replace the missing subpools.  In
the cases where we substituted a subpool it has been clearly
indicated by the stock centers so that anyone can ascertain the
origin of a particular mutant.

     Because we had ~5,000 transformed families to screen in the
fall of 1991, we invited several research groups to help with the
work.  If you would like a list of the participants you may
contact me.  Most of the mutant families identified by these
researchers have been or are being sent to the stock center at
Ohio State as we get them retested.  These mutants will be made
available through both Centers.

     There has been some concern about which mutants were included
in the pooling process.  Several classes were, in fact, excluded. 
The late flowering lines result in larger rosettes so we decided
to keep these out of the pools.  However, all of these have been
sent to the stock centers.  There were 7 primary dwarfs which my
lab is characterizing that were also excluded from the pools. 
However, the vast majority of the >800 putative mutants were
included in the pooling process.  These include:

>60 flower mutants
>60 reduced fertility mutants
>150 embryo-defective mutants
~150 size variants
>50 seedling-lethals
>100 pigment mutants
>100 other various morphological mutants.

     The mutants listed above are either currently available from
the stock center or will be in the next month.

     All of these transformed lines are in the Wassilewskija
background and all were generated with the 3850:1003 Ti plasmid.
Note that the WS accession utilized for these experiments was
maintained for a number of generations in U. S. laboratories, and
a stock of this line is being made available through the stock
centers.  This line should be considered distinct from the Ws of
the Kranz collection.  This new population is described in more
detail elsewhere (Forsthoefel et al. Aust. J. Plant Phys., in
press).

     If you have any specific questions about the population I
would be happy to address them.  Please send questions to my email
address (feldmann at ccit.arizona.edu).

Ken Feldmann
University of Arizona



More information about the Arab-gen mailing list