Compositions and methods for plant genetic modification

Abstract

Mixed duplex oligonucleotides (MDON) are used to effect site-specific genetic alterations in a target DNA sequence of a plant. The MDON are introduced by electroporation into microspores. Thereafter, plants having a desired genetic alteration are produced by germinating the microspores.

1 . A plant microspore comprising a mixed duplex oligonucleotide.  
     
     
         2 . The plant microspore composition of  claim 1  wherein the plant is a member of the family Brassicaceae.  
     
     
         3 . The plant microspore composition of  claim 2  wherein the plant microspore is a  Brassica napus, Brassica rapa, Brassica olercea  or a  Brassica juncea  microspore.  
     
     
         4 . A mutated plant microspore which comprises a genomic mutation wherein the genomic mutation was accomplished by introducing a mixed duplex oligonucleotide into the plant microspore.  
     
     
         5 . The mutated plant microspore of  claim 4  wherein the microspore is from a plant of the family Brassicacaea.  
     
     
         6 . The mutated plant microspore of  claim 5  wherein the plant microspore is a  Brassica napus, Brassica rapa, Brassica oleracea  or  Brassica juncea  microspore.  
     
     
         7 . An isolated plant microspore which comprises a genomic mutation wherein the genomic mutation was accomplished by introducing a mixed duplex oligonucleotide into the plant microspore.  
     
     
         8 . The isolated plant microspore of  claim 7  wherein the microspore is from a plant of the family Brassicacaea.  
     
     
         9 . The isolated plant microspore of  claim 8  wherein the plant microspore is a  Brassica napus, Brassica rapa, Brassica oleracea  or a  Brassica juncea  microspore.  
     
     
         10 . The isolated plant microspore of  claim 7  wherein the microspore is regenerated into a plant that contains the genomic mutation.  
     
     
         11 . A plant regenerated from an isolated microspore of  claim 7  and progeny thereof.  
     
     
         12 . A seed which is derived from the regenerated plant of  claim 11 .  
     
     
         13 . A composition comprising: 
 a. a plant microspore and    b. a mixed duplex oligonucleotide inside the plant microspore wherein the mixed duplex oligonucleotide is capable of causing a genomic mutation in the microspore.    
     
     
         14 . The composition of  claim 13  wherein the microspore is from a plant of the family Brassicacaea.  
     
     
         15 . The composition of  claim 14  wherein the plant microspore is a  Brassica napus, Brassica rapa, Brassica oleracea  or  Brassica juncea  microspore.