Methods of transforming plants

Abstract

Methods for producing homozygous plants, seeds, and plant cells are provided. Also provided are methods of transformation.

What is claimed is:  
     
         1 . A method of transformation comprising stably transforming a plant cell with a polynucleotide of interest and treating the transformed cell with a chromosome doubling agent, wherein the plant cell comprises a haploid somatic cell.  
     
     
         2 . The method of  claim 1  wherein the plant cell is a maize cell.  
     
     
         3 . The method of  claim 1  further comprising providing a growth stimulation protein.  
     
     
         4 . The method of  claim 3  wherein the growth stimulation protein is RepA or Lec 1 .  
     
     
         5 . The method of  claim 1  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         6 . The method of  claim 1  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         7 . The method of  claim 1  further comprising regenerating a homozygous plant from the homozygous plant cell.  
     
     
         8 . A method for producing a homozygous plant comprising: 
 a) stably transforming a plant cell with a polynucleotide of interest, wherein the plant cell comprises a haploid somatic cell;    b) treating the transformed cell with a chromosome doubling agent; and    c) growing the treated cell to produce a homozygous plant.    
     
     
         9 . The method of  claim 8  wherein the plant cell is a maize cell.  
     
     
         10 . The method of  claim 8  further comprising providing a growth stimulation protein.  
     
     
         11 . The method of  claim 10  wherein the growth stimulation protein is RepA or Lec1.  
     
     
         12 . The method of  claim 8  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         13 . The method of  claim 12  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         14 . The method of  claim 8  wherein the polynucleotide of interest is replaced with a second polynucleotide of interest.  
     
     
         15 . A method for producing homozygous seed comprising: 
 a) stably transforming a plant cell with a polynucleotide of interest, wherein the plant cell comprises a haploid somatic cell;    b) growing the transformed cell to produce a regenerated plant;    c) treating the regenerated plant with a chromosome doubling agent; and    d) growing the treated plant to produce homozygous seed.    
     
     
         16 . The method of  claim 15  further comprising growing the homozygous seed to produce a homozygous plant.  
     
     
         17 . The method of  claim 15  wherein the plant cell is a maize cell.  
     
     
         18 . The method of  claim 15  further comprising providing a growth stimulation protein.  
     
     
         19 . The method of  claim 18  wherein the growth stimulation protein is RepA or Lec1.  
     
     
         20 . The method of  claim 15  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         21 . The method of  claim 20  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         22 . The method of  claim 15  wherein the polynucleotide of interest is replaced with a second polynucleotide of interest.  
     
     
         23 . A method for producing a homozygous plant for evaluating recessive genes comprising: 
 a) stably transforming a plant cell with a recessive gene, wherein the plant cell comprises a haploid somatic cell;    b) treating the transformed cell with a chromosome doubling agent; and    c) growing the treated cell to produce a regenerated plant which is homozygous for the recessive gene to be evaluated.    
     
     
         24 . The method of  claim 23  wherein the plant cell is a maize cell.  
     
     
         25 . The method of  claim 23  further comprising providing a growth stimulation protein.  
     
     
         26 . The method of  claim 25  wherein the growth stimulation protein is RepA or Lec1.  
     
     
         27 . The method of  claim 23  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         28 . The method of  claim 27  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         29 . A method for producing a homozygous plant for evaluating knockout genes comprising: 
 a) stably transforming a plant cell with a polynucleotide of interest, wherein the plant cell comprises a haploid somatic cell, and wherein the polynucleotide of interest suppresses expression of a native polynucleotide;    b) treating the transformed cell with a chromosome doubling agent; and    c) growing the treated cell to produce a regenerated plant which is a homozygous knockout for the gene to be evaluated.    
     
     
         30 . The method of  claim 29  wherein the plant cell is a maize cell.  
     
     
         31 . A method for producing a homozygous plant for evaluating lethal versus nonlethal genes comprising: 
 a) stably transforming a plant cell with a polynucleotide of interest, wherein the plant cell comprises a haploid somatic cell;    b) treating the transformed cell with a chromosome doubling agent; and    c) growing the treated cell to produce a regenerated plant which is homozygous for the gene to be evaluated.    
     
     
         32 . The method of  claim 31  wherein the plant cell is a maize cell.  
     
     
         33 . The method of  claim 31  further comprising providing a growth stimulation protein.  
     
     
         34 . The method of  claim 33  wherein the growth stimulation protein is RepA or Lec1.  
     
     
         35 . The method of  claim 31  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         36 . The method of  claim 35  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         37 . A method for producing a homozygous plant containing stacked traits comprising: 
 a) stably transforming a plant cell with one or more polynucleotides of interest, wherein the plant cell comprises a haploid somatic cell, and wherein the one or more polynucleotides of interest produce two or more desired traits;    b) treating the transformed cell with a chromosome doubling agent; and    c) growing the treated cell to produce a regenerated plant which is homozygous for the desired traits.    
     
     
         38 . The method of  claim 37  wherein the plant cell is a maize cell.  
     
     
         39 . The method of  claim 37  further comprising providing a growth stimulation protein.  
     
     
         40 . The method of  claim 39  wherein the growth stimulation protein is RepA or Lec1.  
     
     
         41 . The method of  claim 37  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         42 . The method of  claim 41  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         43 . A method for screening for a transgenic event in an early generation comprising: 
 a) stably transforming a plant cell with a polynucleotide of interest, wherein the plant cell comprises a haploid somatic cell;    b) treating the transformed cell with a chromosome doubling agent;    c) growing the treated cell to produce a regenerated homozygous plant; and    d) screening for the transgenic event.    
     
     
         44 . The method of  claim 43  wherein the plant cell is a maize cell.  
     
     
         45 . The method of  claim 43  further comprising providing a growth stimulation protein.  
     
     
         46 . The method of  claim 44  wherein the growth stimulation protein is RepA or Lec1.  
     
     
         47 . The method of  claim 43  wherein the plant cell is a haploid embryo cell produced by crossing a selected female parent with a male inducer line.  
     
     
         48 . The method of  claim 47  wherein the inducer line contains a scorable marker gene operably linked to an embryo-preferred promoter.  
     
     
         49 . A method for obtaining homozygous seed comprising: 
 a) crossing a selected female parent with a male inducer line to produce a haploid embryo cell;    b) providing a growth stimulation protein;    c) treating the haploid embryo cell with a chromosome doubling agent;    d) growing the treated embryo cell to produce a regenerated plant having homozygous seed.    
     
     
         50 . A method for obtaining homozygous seed comprising: 
 a) crossing a selected female parent with a male inducer line to produce a haploid embryo;    b) providing a growth stimulation protein;    c) treating the haploid embryo with a chromosome doubling agent;    d) growing the treated embryo to produce a regenerated plant having homozygous seed.    
     
     
         51 . A method for obtaining homozygous seed comprising: 
 a) crossing a selected female parent with a male inducer line to produce a haploid seed;    b) providing a growth stimulation protein;    c) treating the haploid seed with a chromosome doubling agent; and    d) growing the treated seed to produce a plant having homozygous seed.    
     
     
         52 . A method for obtaining homozygous seed comprising: 
 a) crossing a selected female parent with a male inducer line to produce a haploid seed;    b) providing a growth stimulation protein;    c) growing the seed to produce a haploid seedling;    d) treating the haploid seedling with a chromosome doubling agent; and    e) growing the treated seedling to produce a plant having homozygous seed.    
     
     
         53 . A method for obtaining haploid embryos comprising crossing a selected female parent with a male inducer line and providing a growth stimulation protein to produce haploid embryos.  
     
     
         54 . A method for obtaining haploid seeds comprising crossing a selected female parent with a male inducer line and providing a growth stimulation protein to produce haploid seeds.  
     
     
         55 . A method for increasing chromosome doubling in a haploid cell comprising providing a growth stimulation protein to a haploid cell and treating the haploid cell with a chromosome doubling agent.