Process for the incorporation of foreign dna into the genome of dicotyledonous plants

• US 4,940,838 A
• Filed: 02/23/1984
• Issued: 07/10/1990
• Est. Priority Date: 02/24/1983
• Status: Expired due to Term

• Current Assignee: Syngenta Mogen BV (China National Chemical Corporation)
• Original Assignee: Andreas Hoekema, Paul J.J. Hooykaas, Robbert A. Schilperoort
• Inventors: Hoekema, Andreas, Hooykaas, Paul J. J., Schilperoort, Robbert A.
• Primary Examiner(s): Tarcza, J. E.
• Application Number:
  US06/583,022
• Time in Patent Office: 2,329 Days
• Field of Search: 435/172.3, 435/240, 435/317, 935/30, 935/35, 935/56, 935/67
• US Class Current: 800/294
• CPC Class Codes: C12N 15/8205 Agrobacterium mediated transformation

Abstract

The invention relates to a process for the incorporation of foreign DNA into chromosomes of dicotyledonous plants by infecting the plants or incubating plant protoplasts with Agrobacterium bacteria, which contain one or more plasmids, wherein bacteria are used which contain at least one plasmid having the vir-region of a Ti (tumour inducing) plasmid but no T-region, and at least one other plasmid having a T-region with incorporated therein foreign DNA but no vir-region, as well as to a Agrobacterium bacteria wherein at least one plasmid which has the vir-region of a Ti (tumour inducing) plasmid but no T-region and at least one other plasmid which has a wild type T-region with incorporated in it foreign DNA but no vir-region.

Claims

1. A process for the incorporation of foreign DNA into chromosomes of dicotyledonous plants, comprising infecting the plants or incubating plant protoplasts with Agrobacterium bacteria, which contain plasmids, said Agrobacterium bacteria containing at least one plasmid having the vir-region of a Ti plasmid but no T-region, and at least one other plasmid having a T-region with incorporated therein foreign DNA but no vir-region.

9. A process as in claim 1, wherein said Agrobacterium bacteria contains one plasmid having the vir-region of a Ti-plasmid but no T-region and one plasmid having a T-region with incorporated therein foreign DNA but no vir-region.

10. A process as in claim 1, further comprising regenerating the plants from the plant protoplast after infecting said protoplasts with said agrobacterium containing at least one plasmid having the vir-region of a Ti-plasmid but no T-region and at least one other plasmid having a T-region with incorporated therein foreign DNA bur no vir-region.

11. A process as in claim 1 wherein said T region with foreign DNA and no vir region incorporated in it is free of ONC-genes.

12. Plants and plant cells obtained after the genetic properties of the original plants or plant cells have been modified with application of the process according to claim 11.

2. Agrobacterium bacteria, comprising at least one plasmid which has the vir-region of a Ti plasmid but no T-region and at least one other plasmid which has a T-region with foreign DNA incorporated in it but no vir-region.

3. Agrobacterium bacteria, as in claim 2, said T-region with foreign DNA incorporated in it is

an artificial T-region.

4. Agrobacterium bacteria, as in claim 3, said artificial T-region with foreign DNA and no vir-region incorporated in it, is free of onc-genes.

5. Agrobacterium bacteria, as in claim 2, whereby said T-region with foreign DNA incorporated in it but no vir-region is a wild-type T-region.

6. Agrobacterium bacteria, as in claim 2, wherein said T-region comprises the foreign DNA introduced in between wild-type T-region border sequences.

7. A bacteria as in claim 6, wherein said border sequences comprise a special base sequence of about 23 base pairs located at the wild-type T-region.

8. Agrobacteria bacteria as in claim 2, wherein said Agrobacterium bacteria contains one plasmid having the vir-region of a Ti-plasmid but no T-region and one plasmid having a T-region with incorporated therein foreign DNA but no vir-region.

13. A process for the production of Agrobacterium bacteria which contains at least one plasmid which has the vir-region of a Ti-plasmid but no T-region and at least one other plasmid which has a T-region with foreign DNA incorporated in it but no vir-region, comprising:

(a) using Escherichia coli as a host and incorporating foreign DNA into a plasmid therein which contains a T-region and a replicator having a broad bacterial host range

(b) introducing the resulting plasmid into Agrobacterium bacteria which contain at least one plasmid which has the vir-region of a Ti-plasmid but no T-region.