Vacuum Infiltration of seedling plants

A method for direct plant transformation using seedlings and Agrobacterium comprising: A) contacting at least one seedling with Agrobacterium cells, said Agrobacterium cells harboring a vector, said vector enabling said Agrobacterium cells to transfer T-DNA containing at least one gene or gene fragment to said seedling;
B) applying a vacuum to said seedling in contact with said Agrobacterium cells at a first time, said vacuum of sufficient strength to force said Agrobacterium cells into intimate contact with said seedling such that said Agrobacterium cells transfer said T-DNA to cells of said seedling at a second time, wherein said first and second time are the same or different.

A method for direct plant transformation using seedlings and Agrobacterium comprising: A) contacting at least one seedling with a mixture of Agrobacterium cells, said mixture comprising cells from a Agrobacterium strain harboring a vector with a DNA fragment and cells from said Agrobacterium strain harboring said vector a second DNA fragment, said vector enabling said Agrobacterium cells to transfer said T-DNA to said seedling;
B) applying a vacuum to said seedling in contact with said Agrobacterium cells at a first time, said vacuum of sufficient strength to force said Agrobacterium cells into intimate contact with said seedling such that said Agrobacterium cells transfer T-DNA to cells of said seedling at a second time, wherein said first and second time are the same or different.

A method for direct plant transformation using seedlings and Agrobacterium comprising: A) contacting at least one seedling with Agrobacterium cells, said Agrobacterium cells harboring a vector, said vector enabling said Agrobacterium cells to transfer T-DNA containing at least one gene or gene fragment and a selectable marker gene to said seedling;
B) applying a vacuum to said seedling in contact with said Agrobacterium cells at a first time, said vacuum of sufficient strength to force said Agrobacterium cells into intimate contact with said seedling such that said Agrobacterium cells transfer said T-DNA to cells of said seedling at a second time, wherein said first and second time are the same or different;
C) allowing said transformed seedling to grow to maturity and set seed;
D) germinating said seed to form progeny;
E) exposing said progeny to an agent enabling detection of selectable marker gene expression;
F) selecting for progeny expressing said selectable marker gene and at least one gene, said expression of said selectable marker gene and at least one gene indicating gene transfer.