Methods for enhancing plant transformation frequencies

Abstract

The invention provides improved transformation methods. In particular the method provides increased transformation frequency, especially in recalcitrant plants. The method comprises stably transforming a target cell with at least one polynucleotide of interest. The target cell has been previously transformed to stimulate growth of the cell and has gone through at least one cell division.

What is claimed is:  
     
         1 . A method for transforming a target plant cell comprising stably transforming the target cell with a growth stimulation polynucleotide to produce a modified cell, growing the modified cell through at least one cell division to produce a progeny cell expressing the growth stimulation polynucleotide, and then transforming the progeny cell with at least one polynucleotide of interest, wherein the growth stimulation polynucleotide is an anti-apoptosis polynucleotide other than baculovirus p35 or baculovirus iap; a hormone polynucleotide; or a silencing construct targeted against cell cycle repressors.  
     
     
         2 . The method of  claim 1  wherein transformation frequency is increased compared to corresponding plant cells that do not contain the growth stimulation polynucleotide.  
     
     
         3 . The method of  claim 1  wherein the progeny cell is from T0 transgenic cultures, regenerated plants or any subsequent progeny cell expressing the growth stimulation polynucleotide.  
     
     
         4 . The method of  claim 3  wherein the progeny cell is from a T0 regenerated plant or a plant from any subsequent generation expressing the growth stimulation polynucleotide.  
     
     
         5 . The method of  claim 1  wherein the growth stimulation polynucleotide is an anti-apoptosis polynucleotide other than baculovirus p35 or baculovirus iap.  
     
     
         6 . The method of  claim 1  wherein the growth stimulation polynucleotide is a hormone polynucleotide.  
     
     
         7 . The method of  claim 1  wherein the growth stimulation polynucleotide is a silencing construct targeted against cell cycle repressors.  
     
     
         8 . The method of  claim 1  wherein the target cell is from a monocot or a dicot plant.  
     
     
         9 . The method of  claim 8  wherein the target cell is from a monocot.  
     
     
         10 . The method of  claim 9  wherein the target cell is from a maize plant.  
     
     
         11 . The method of  claim 8  wherein the target cell is from a dicot.  
     
     
         12 . The method of  claim 11  wherein the target cell is from a soybean plant.  
     
     
         13 . The method of  claim 8  wherein the target cell is from corn, soybean, sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley, potato, tomato, or millet.  
     
     
         14 . A method of transformation comprising transforming a target plant cell with at least one polynucleotide of interest, wherein the target cell has been previously stably transformed to express a growth stimulation polynucleotide and has gone through at least one cell division, wherein the growth stimulation polynucleotide is an anti-apoptosis polynucleotide other than baculovirus p35 or baculovirus iap; a hormone polynucleotide; or a silencing construct targeted against cell cycle repressors.  
     
     
         15 . The method of  claim 14  wherein the target cell is from T0 transgenic cultures, regenerated plants or any subsequent progeny expressing the transgenic growth stimulation polynucleotide.  
     
     
         16 . The method of  claim 15  wherein the target cell is from a T0 regenerated plant or any subsequent progeny expressing the transgenic growth stimulation polynucleotide.  
     
     
         17 . The method of  claim 14  wherein the growth stimulation polynucleotide is an anti-apoptosis polynucleotide other than baculovirus p35 or baculovirus iap.  
     
     
         18 . The method of  claim 14  wherein the growth stimulation polynucleotide is a hormone polynucleotide.  
     
     
         19 . The method of  claim 14  wherein the growth stimulation polynucleotide is a silencing construct targeted against cell cycle repressors.  
     
     
         20 . The method of  claim 14  wherein the target cell is from a monocot or a dicot plant.  
     
     
         21 . The method of  claim 20  wherein the target cell is from a monocot.  
     
     
         22 . The method of  claim 21  wherein the target cell is from a maize plant.  
     
     
         23 . The method of  claim 20  wherein the target cell is from a dicot.  
     
     
         24 . The method of  claim 23  wherein the target cell is from a soybean plant.  
     
     
         25 . The method of  claim 20  wherein the target cell is from corn, soybean, sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley, potato, tomato, or millet.