Delayed fruit deterioration allele in plants and methods of detection

Abstract

The present invention relates to an isolated nucleic acid molecule comprising a nucleotide sequence at least 90% identical to a nucleotide sequence encoding a protein having the amino acid sequence of SEQ ID NO:4, where the protein as an aspartic acid (Asp) at amino acid position 106. The present invention also relates to a transgenic plant with reduced or delayed softening and deterioration of fruit compared to wild-type plants, methods of genetic screening for plants having reduced or delayed softening and deterioration of fruit, isolated oligonucleotides, and a method of imparting to a plant the dfd trait.

What is claimed: 
     
         1 . A method of genetic screening for plants having reduced or delayed softening and deterioration of fruit, said method comprising:
 identifying a plant or plant cell containing a NOR gene, wherein the NOR gene comprises a nucleic acid molecule encoding the amino acid sequence of SEQ ID NO:4.   
     
     
         2 . The method according to  claim 1 , wherein said identifying is carried out by a procedure selected from DNA sequencing, PCR, single base pair extension assay, ligase detection reaction, and ligase chain reaction. 
     
     
         3 . The method according to  claim 2 , wherein said identifying is carried out by the single base pair extension assay, the assay comprising template-directed dye-terminator incorporation and fluorescence polarization detection (FP-TDI). 
     
     
         4 . An isolated oligonucleotide comprising a nucleotide sequence selected from the group consisting of: 
       
         
           
                 
                 
               
                     
                   Nor-F 
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   5′-ACTCATCGTCCACTACCTCA-3′ 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   Nor1556 
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   5′-TGTCGAACCTCTTTCGGCTA-3′. 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         5 . A kit for detecting a NOR gene in a plant, said kit comprising an oligonucleotide comprising the nucleotide sequence of: 
       
         
           
                 
                 
               
                     
                   Nor-F 
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   5′-ACTCATCGTCCACTACCTCA-3′ 
                 
                     
                   or 
                 
                     
                     
                 
                     
                   Nor1556 
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   5′-TGTCGAACCTCTTTCGGCTA-3′. 
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         6 . The kit according to  claim 5  further comprising:
 materials to perform PCR; 
 one or more buffers; and 
 directions for using the kit. 
 
     
     
         7 . A method of genetic screening comprising:
 identifying a nucleotide sequence linked to a NOR gene, wherein said NOR gene comprises a nucleic acid molecule encoding the amino acid sequence of SEQ ID NO:4, and   utilizing said nucleotide sequence as a marker to screen or map said NOR gene.   
     
     
         8 . The method according to  claim 7 , wherein said identifying is carried out by a procedure selected from DNA sequencing, PCR, single base pair extension assay, ligase detection reaction, and ligase chain reaction. 
     
     
         9 . The method according to  claim 8 , wherein said identifying is carried out by the single base pair extension assay, the assay comprising template-directed dye-terminator incorporation and fluorescence polarization detection (FP-TDI).