Agrobacterium for transient transfection of whole plants
Inventors
Roemer, Patrick • Bortesi, Luisa • Tiede, Doreen • Giritch, Anatoli • Gleba, Yuri
Assignees
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Publication Number
US-10934553-B2
Publication Date
2021-03-02
Expiration Date
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.
Core Innovation
The invention relates to Agrobacterium strain CryX having accession no. DSM25686, and derivatives that can be used for transient transfection/transient expression in a Nicotiana plant or leaves. CryX is described as an Agrobacterium strain for in planta transient delivery, including use with binary T-DNA vectors.
The disclosure further characterizes that a vir plasmid used with CryX is disarmed (non-oncogenic). Binary vector arrangements are described with a virG gene, including a virG gene encoding VirG protein from Agrobacterium tumefaciens strain LBA4404 (SEQ ID NO:1) or a virG N54D mutant.
A key aspect is the improved transient transfection efficiency obtainable with strain CryX compared with strain KYRT1, as evaluated in Nicotiana benthamiana leaves. The described evaluation compares GFP fluorescence intensity after infiltration using a needleless syringe.
Claims Coverage
The provided independent claims are clm-00001 and clm-00002. Across these claims, the core inventive concept is the use of Agrobacterium strain CryX (accession DSM25686) to achieve higher transient transfection/transient expression efficiency than KYRT1, in Nicotiana benthamiana leaves, with GFP fluorescence intensity comparison.
CryX-based transient transfection with higher efficiency than KYRT1
A process of transiently transfecting a Nicotiana plant or leaves by contacting with an Agrobacterium suspension comprising Agrobacterium cells of strain CryX having accession no. DSM25686, wherein the transient transfection efficiency obtainable with strain CryX is higher than with strain KYRT1.
CryX-based transient expression with higher efficiency than KYRT1
A process of transiently expressing a DNA sequence of interest in a Nicotiana plant by contacting the plant or leaves on the plant with an Agrobacterium suspension comprising Agrobacterium cells of strain CryX having accession no. DSM25686, wherein the transient transfection efficiency obtainable with strain CryX is higher than with strain KYRT1.
Overall, the claim coverage centers on using Agrobacterium strain CryX (DSM25686) to contact Nicotiana plant material for in planta transient delivery, where CryX provides higher transient transfection efficiency than KYRT1 as assessed by GFP fluorescence intensity.
Stated Advantages
Increased transient transfection efficiency obtainable with strain CryX compared with strain KYRT1.
Documented Applications
Transiently transfecting Nicotiana plant or leaves by contacting with Agrobacterium suspension of strain CryX and evaluating GFP fluorescence intensity after infiltration.
Transiently expressing a DNA sequence of interest in a Nicotiana plant by contacting with Agrobacterium suspension of strain CryX and evaluating GFP fluorescence intensity after infiltration.
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