Markerless transformation

Abstract

Methods for identification of successful transformation without a need for in vitro selection are provided. Direct detection of nucleotide sequences of interest is described which eliminate the need for use of ancillary nucleotide sequences.

I claim:  
     
         1 . A method for producing a transgenic plant that includes a nucleotide sequence of interest and is free of ancillary nucleic acid sequences, where said method comprises: 
 a) transforming a plant tissue or plant cell with a gene of interest;    b) regenerating said transformed plant tissue or plant cell to produce a new plant; and    c) directly detecting the presence or absence in said regenerated plant of the nucleotide sequence, or encoded product of said nucleotide sequence, of interest.    
     
     
         2 . The method of  claim 1 , wherein the direct detection method is selected from the group consisting of southern hybridization, polymerase chain reaction, western hybridization or the gene product of interest and HPLC for metabolic product of the protein product.  
     
     
         3 . The method of  claim 2  wherein the direct detection method is southern hybridization and HPLC for metabolic product of the protein product.  
     
     
         4 . The method of  claim 2  wherein the direct detection method is polymerase chain reaction and HPLC for metabolic product of the protein product.  
     
     
         5 . The method of  claim 2  wherein the direct detection method is western hybridization and HPLC for metabolic product of the protein product.  
     
     
         6 . The method of  claim 2  wherein the direct detection method is the gene product of interest and HPLC for metabolic product of the protein product.  
     
     
         7 . A plant transformed by the method of  claim 1 .  
     
     
         8 . Seed and progeny of the plant of  claim 7.