Agrobacterium for transient transfection of whole plants

Abstract

A process of transiently transfecting a plant or leaves on a plant, comprising contacting said plant or said leaves with a suspension comprising Agrobacterium cells of strain CryX or a derivative strain of strain CryX, wherein said derivative strain has the chromosomal background of strain CryX or said derivative strain contains the vir plasmid of strain CryX or a derivative of said vir plasmid.

1 . A process of transiently transfecting a plant or leaves on a plant, comprising contacting said plant or said leaves with a suspension comprising  Agrobacterium  cells of strain CryX or a derivative strain of strain CryX,
 wherein said derivative strain has the chromosomal background of strain CryX or said derivative strain contains the vir plasmid of strain CryX or a derivative of said vir plasmid.   
     
     
         2 . A process of transiently expressing a DNA sequence of interest in a plant, comprising contacting said plant or said leaves on said plant with a suspension comprising  Agrobacterium  cells of strain CryX or a derivative strain of strain CryX,
 wherein said derivative strain has the chromosomal background of strain CryX or said derivative strain contains the vir plasmid of strain CryX or a derivative of said vir plasmid.   
     
     
         3 . The process according to  claim 1 , wherein said strain CryX or said derivative strain contains a binary vector having T-DNA comprising a DNA sequence of interest to be transfected into cells of said plant or leaves. 
     
     
         4 . The process according to  claim 1 , wherein said strain CryX or said derivative strain contains a disarmed vir plasmid or disarmed Ti-plasmid. 
     
     
         5 . The process according to  claim 1 , wherein said derivative strain of strain CryX contains a vir plasmid that is a derivative of the vir plasmid of strain CryX, and said derivative strain achieves at least 70% of the T-DNA transfer efficiency from a T-DNA-containing binary vector into plant cells of the T-DNA transfer efficiency with strain CryX. 
     
     
         6 . The process according to  claim 1 , wherein said derivative of the vir plasmid of strain CryX encodes the virG protein of strain CryX. 
     
     
         7 . The process according to  claim 6 , wherein said derivative of the vir plasmid of strain CryX contains the vir region of the vir plasmid of strain CryX. 
     
     
         8 . The process according to  claim 3 , wherein said binary vector comprises a virG gene expressible in said strain CryX or said derivative strain. 
     
     
         9 . The process according to  claim 8 , wherein said virG gene encodes a VirG protein from  Agrobacterium tumefaciens  strain LBA4404 of SEQ ID NO: 1, or is an N54D mutant of the VirG protein encoded by the virG gene from  A. tumefaciens  strain LBA4404. 
     
     
         10 . The process according to  claim 1 , wherein said derivative of the vir plasmid of strain CryX is obtained by introducing a T-DNA of interest into the vir plasmid of strain CryX. 
     
     
         11 . The process according to  claim 1 , wherein said plant or said leaves on a plant are contacted with said suspension by spraying or by vacuum infiltrating said plant or said leaves on a plant with said suspension. 
     
     
         12 . The process according to  claim 1 , wherein said plant is a dicot plant such as  Nicotiana benthamiana,  tobacco, cotton, soybean, rapeseed, pepper, potato, tomato, or a monocot plant. 
     
     
         13 . The process according to  claim 1 , wherein said  Agrobacterium  strain CryX is the strain having DSM accession No: DSM25686. 
     
     
         14 .  Agrobacterium  strain CryX having DSM accession No: DSM25686 or a derivative strain of strain CryX, wherein said derivative strain has the chromosomal background of strain CryX, or said derivative strain contains the vir plasmid of strain CryX or a derivative of said vir plasmid; or an  Agrobacterium  cell of strain CryX or of said derivative strain. 
     
     
         15 . The  Agrobacterium  cell according to  claim 14 , wherein said cell further comprises a binary vector comprising a T-DNA comprising a DNA sequence of interest to be transfected into a plant. 
     
     
         16 . A kit comprising
 an  Agrobacterium  cell of said strain or said derivative strain as defined in  claim 14  and   a vector containing in T-DNA a DNA sequence of interest to be transfected into cells of a plant.   
     
     
         17 . Vir plasmid of  Agrobacterium  strain CryX having DSM accession No: DSM25686. 
     
     
         18 .  Agrobacterium  cells having the chromosome of strain CryX having DSM accession No: DSM25686. 
     
     
         19 . Aqueous cell suspension of  Agrobacterium  strain CryX having DSM accession No: DSM25686 or a derivative strain of strain CryX, said suspension having a cell concentration of at most 1.1·10 6  cfu/ml of the suspension, preferably at most 4.4·10 5  cfu/ml of the suspension, and more preferably of at most 1.1·10 5  cfu/ml of the suspension.