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The European Arabidopsis Stock Centre

Feldmann T-DNA

Donated by

  • Ken Feldmann Department of Plant Sciences, College of Agricultural and Life Sciences, University of Arizona

Click here to view all 830 of these lines.

Description

Sets in this collection

Nasc code Description Set contents
N3115 Complete set of Feldmann 100 pools - 1st set View set contents
N3116 Complete set of Feldmann 20 pools - 1st set View set contents
N6400 Complete set of Feldmann 20 pools - 2nd set View set contents
N6500 Complete set of Feldmann 100 pools - 2nd set View set contents
N6501 Complete set of Feldmann 325 pools of 20 - 1st & 2nd set View set contents
N6502 Complete set of Feldmann 65 pools of 100 - 1st & 2nd set View set contents
N84441 Complete set of Feldmann 400 pools of 10 - set 3 View set contents
N84442 Complete set of Feldmann 100 pools - set 3 View set contents

Feldmann T-DNA Tagged lines from seed transformation

T-DNA Transformation scheme:

  • Imbibe Ws seed and cocultivate with Agrobacterium = T1
  • Allow T1 to self and collect seed = T2
  • Test T2 seeds on Kanamycin and transfer KanR to soil
  • Allow to self and collect seed = T3
  • Test T3 seeds on Kanamycin for number of functional inserts and screen on agar and soil for alterations in phenotype.

Pooling of the lines

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 see below. 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. All of these transformed lines are in the Wassilewskija background (N1601) 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 new population is described in more detail elsewhere (Forsthoefel, N.R. et al. 1992).

If you have any specific questions about the population I would be happy to address them. Please send questions to my e-mail address:feldmann@ccit.arizona.edu

NOTE: The following individuals have participated in the screening of individual mutants:

  • Embryo defectives: Robert Goldberg, John Harada and David Meinke (available through ABRC)
  • Flower: Elliot Meyerowitz and Patricia Zambryski
  • Early development (to be received later): June Medford
  • Reduced fertility: Robert Fischer and Robert Goldberg