Agrobacterium-mediated transformation

[EnnaB]

I'm by no means a molecular biologist so please excuse any mistakes.

I'm interested in agrobacterium-mediated transformation of a plant using the pNOV2819 plasmid (pic below) which uses the pmi selectable marker. I read a paper where its efficacy in transforming almonds was tested (https://www.ncbi.nlm.nih.gov/pubmed/16534597). However, no new gene was inserted into the plasmid so the transformed plants only contained the selectable marker (and promoter/terminator). I wondered if a new gene was inserted (for example at the AscI restriction site) how the selection would work in agrobacterium and the plant in terms of growth on a mannose medium and PCR/Southern blotting to confirm transgene incorporation.

Any advice/explanation would be great!

Edited June 18, 2019 by EnnaB
submitted before finished by accident

[CharonY]

Essentially you would first screen spectinomycin (i.e. plasmid carrying) bacteria and confirm insertion via Southern (though a nested PCR would also work). From the looks of it, there is no marker for counter-selection, i.e. you would just have to screen clones, unless I am missing something. The explants can then be selected via their ability to grow with mannose as sole carbon source. Typically you would then confirm the insertion in plants again via PCR (and Southern, if you feel thorough).