Rhizobia transformants which symbiotically fixes nitrogen in non-legumes, a material for treating seeds of a non-legume plant, non-legume seeds, a non-legume plant and a method for producing rhizobia transconjungants

• US 5,229,291 A
• Filed: 12/11/1990
• Issued: 07/20/1993
• Est. Priority Date: 12/30/1985
• Status: Expired due to Fees

• Current Assignee: Novo Industri A/S (Novo Nordisk A/S)
• Original Assignee: Novo Industri A/S (Novo Nordisk A/S)
• Inventors: Sorensen, Grete Morch, Nielsen, Sven-Erik
• Primary Examiner(s): Fox, David T.
• Application Number:
  US07/626,233
• Time in Patent Office: 952 Days
• Field of Search: 435/34, 435/172.1, 435/172.2, 435/172.3, 435/244, 435/252.2, 435/252.3, 435/317.1, 435/878, 47/58, 71/7
• US Class Current: 435/252.2
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 15/00 Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00
• CPC Class Codes: C12R 2001/41 Rhizobium
• CPC Class Codes: Y10S 435/878 Rhizobium

Abstract

Rhizobia transformants that nodulate and fix nitrogen in non-legumes. The nodulated non-legume plants can be grown without nitrogenous fertilizer and have at least the same or higher protein content, dry matter content and nitrogen content than their non-nodulated counterparts which are fertilized by the addition of nitrogenous fertilizer. The straw remaining after harvesting the nodulated non-legumes is also high in protein content.

Claims

1. A Rhizobium transconjugant which fixes nitrogen in wheat plants as deposited with the ATCC and assigned accession number 53,407.

7. The Rhizobium transconjugant of claim 1, 2, 3, 4, 5, or 6 characterized a forming snowy white colonies before entering symbiosis with a non-legume host partner when the transconjugant is cultured on a nutrient medium containing, in addition to nutrients required for growth, an extract of each legume host partner of each parent Rhizobium species of the Rhizobium transconjugant.

8. The Rhizobium transconjugant of claim 1, 2, 3, 4, 5, or 6 characterized a forming grey colonies after entering symbiosis with a non-legume host partner when the transconjugant is cultured on a nutrient medium containing, in addition to nutrients required for growth, an extract of each legume host partner of each parent Rhizobium species of the Rhizobium transconjugant and an extract of the non-legume host partner of the Rhizobium transconjugant.

15. A method for producing the Rhizobia F₂ transconjugants of claim 1, 2, 3, 4, 5, or 6 that symbiotically fix nitrogen in non-legumes, comprising:

(a) streaking a first Rhizobium parent and a second Rhizobium patent in alternating rows on a solid nutrient medium containing in addition to nutrient essential for growth, a non-denatured shoot extract of a legume host partner of each parent Rhizobium;

(b) culturing the Rhizobia parents at a temperature of about 18°

C. to about 30°

C. so that the Rhizobia parents form colonies having a specific color;

(c) selecting a Rhizobium F₁ transconjugant milky-white colony that grows in between the Rhizobia parent colonies;

(d) streaking the Rhizobium F₁ transconjugant in alternating rows with a third Rhizobium parent on the nutrient medium of step (a) further comprising a legume shoot extract of a legume host partner of the third Rhizobia parent;

(e) culturing the Rhizobium F₁ transconjugant and the third Rhizobium parent at a temperature of about 18°

C. to about 30°

C. so that the Rhizobia form colonies; and

(f) selecting a Rhizobium F₂ transconjugant snowy white colony that grows in between the Rhizobia F₁ transconjugant and the third Rhizobia parent.

16. The method according to claim 15 in which the third Rhizobia parent comprises a third Rhizobia species, a second Rhizobia F₁ transconjugant or a second Rhizobia F₂ transconjugant.

2. A Rhizobium transconjugant which fixes nitrogen in barley plants as deposited with the ATCC and assigned accession number 53,404.

9. The Rhizobium transconjugant of claim 2 which symbiotically fixes nitrogen in the barley strain Hasso.

10. The Rhizobium transconjugant of claim 2 which symbiotically fixes nitrogen in the barley strain Cerise.

11. The Rhizobium transconjugant of claim 2 which symbiotically fixes nitrogen in the barley strain Harry.

12. The Rhizobium transconjugant of claim 2 which symbiotically fixes nitrogen in the barley strain Igri.

3. A Rhizobium transconjugant which fixes nitrogen in sorghum plants as deposited with the ATCC and assigned accession number 53,405.

4. A Rhizobium transconjugant which fixes nitrogen in rice plants as deposited with the ATCC and assigned accession number 53,406.

5. A Rhizobium transconjugant which fixes nitrogen in Brassica plants as deposited with the ATCC and assigned accession number 53,564.

6. A Rhizobium transconjugant which fixes nitrogen in the vicinity of eucalyptus roots as deposited with the ATCC and assigned accession number 53,562.

13. The Rhizobium transconjugant of claim 6 which is capable of acting as a nitrogen fertilizer for a non-legume belonging to the family Myrtaceae.

14. The Rhizobium transconjugant of claim 13 which is capable of acting as a nitrogen fertilizer for Eucalyptus.