Vacuum Infiltration of flowering plants

A method for direct plant transformation using plants and Agrobacterium comprising: A) contacting the aerial portions of at least one plant at the time of flowering 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 plant; and
B) applying a vacuum to said plant portions 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 plant such that said Agrobacterium cells transfer said T-DNA to cells of said plant at a second time to form a transformed plant, wherein said first time and said second time are the same or different.

A method for direct transformation of a plant comprising: A) vernalizing and germinating initial seed to form said plant contacting the aerial portions of said plant at the time of flowering 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 plant; and
B) applying a vacuum to said plant portions 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 plant such that said Agrobacterium cells transfer said T-DNA to cells of said plant at a second time to form a transformed plant, wherein said first time and said second time are the same or different.

A method for direct plant transformation using plants at the time of flowering and Agrobacterium comprising:
A) contacting aerial portions of at least one plant at the time of flowering 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 with a second DNA fragment, said vector enabling said Agrobacterium cells to transfer said T-DNA to said plant; and
B) applying a vacuum to said plant portions 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 plant such that said Agrobacterium cells transfer T-DNA to cells of said plant at a second time to form a transformed plant, wherein said first time and said second time are the same or different.

A method for direct transformation of a plant at the time of flowering comprising: A) vernalizing and germinating initial seed to form said plant contacting aerial portions of said plant at the time of flowering 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 with a second DNA fragment, said vector enabling said Agrobacterium cells to transfer said T-DNA to said plant; and
B) applying a vacuum to said plant portions 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 plant such that said Agrobacterium cells transfer T-DNA to cells of said plant at a second time to form a transformed plant, wherein said first time and said second time are the same or different.

A method for direct plant transformation using plants at the time of flowering and Agrobacterium comprising: A) contacting aerial portions of at least one plant at the time of flowering 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 plant;
B) applying a vacuum to said plant portions 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 plant such that said Agrobacterium cells transfer said T-DNA to cells of said plant at a second time to form a transformed plant, wherein said first time and said second time are the same or different;
C) allowing said transformed plant 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; and
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.

A method for direct transformation of a plant at the time of flowering comprising: A) vernalizing and germinating initial seed to form said plant;
B) contacting aerial portions of said plant at the time of flowering 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 plant;
C) applying a vacuum to said plant portions 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 plant such that said Agrobacterium cells transfer said T-DNA to cells of said plant at a second time to form a transformed plant, wherein said first time and said second time are the same or different;
D) allowing said transformed plant to grow to maturity and set seed; germinating said seed to form progeny;
E) exposing said progeny to an agent enabling detection of selectable marker gene expression; and
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.