I am planning to transform Arabidopsis plants from several different accessions using the floral dip method. To minimize the risk of cross-contamination or potential cross-pollination between accessions, I'm wondering whether it's necessary to prepare and use separate Agrobacterium infiltration solutions for each genotype.
Has anyone encountered issues with cross-contamination or misidentification of transformants when using a shared infiltration solution across accessions?
Are there best practices for handling multiple accessions during floral dipping to maintain genetic integrity and ensure clear downstream identification of transformants?
Any insights or advice would be greatly appreciated!
It's probably not a big risk. The buds that are the best stage for Agro infiltration are too young for pollination so even if there is pollen in the solution (and it is transferred to a stigma, and the pollen germinates, and leads to a seed) the odds of cross-pollinated seed also being transgenic are very low. Still, it's a good idea in general to have nice fresh dipping solution for each batch of plants.
If you want to both decrease the odds of a cross and increase the % of transformed seeds out of total seeds (and you have lots of time on your hands) you can clip off all open flowers and seed pods prior to the Agro dip.
You are more likely to cross pollination offspring by having different ecotypes in the same growth area.
Flower Bud Dipping or Vacuum Infiltration—Two Methods of Arabidopsis thaliana Transformation
Amy Klocko I really appreciate your quick reply! Thank you very much for your detailed and thoughtful response!
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