Fungal resistant plants expressing hydrophobin

Patent metadata

Field	Value
Publication number	US-10344296-B2
Application number	US-201314390574-A
Country	US
Kind code	B2
Filing date	Mar 15, 2013
Priority date	Apr 5, 2012
Publication date	Jul 9, 2019
Grant date	Jul 9, 2019

Abstract

The present invention relates to a method of increasing resistance against fungal pathogens of the family Phacopsoraceae plants and/or plant cells. This is achieved for instance by increasing the expression of a hydrophobin protein or fragment thereof in a plant, plant part and/or plant cell in comparison to wild type plants, wild type plant parts and/or wild type plant cells. In the transgenic plants hydrophobin can be expressed as a fusion protein to facilitate and/or enhance expression. Furthermore, the hydrophobin protein can be expressed including a secretion signal sequence which mediates secretion of the protein into the apoplast and/or into the cuticule.

First claim

The invention claimed is:

1. A method for preventing, reducing, or delaying Phakopsora infection in a transgenic soybean plant, a transgenic soybean plant part, or a transgenic soybean plant cell, the method comprising:
providing a transgenic soybean plant, a transgenic soybean plant part, or a transgenic soybean plant cell with an exogenous nucleic acid encoding a hydrophobin protein comprising an amino acid sequence with at least 90% identity to SEQ ID NO: 42 over the entire length of SEQ ID NO: 42, wherein the hydrophobin protein confers increased resistance against Phakopsora thereto in comparison to a wild type soybean plant, wild type soybean plant part, or wild type soybean plant cell, and
growing the transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell in the presence of a fungal pathogen of the genus Phakopsora , wherein Phakopsora infection is prevented, reduced, or delayed in the transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell as compared to a wild type soybean plant, wild type soybean plant part, or wild type soybean plant cell.

2. The method of claim 1 , wherein the exogenous nucleic acid is in functional linkage with a promoter.

3. The method of claim 2 , wherein the exogenous nucleic acid encoding a hydrophobin protein encodes a hydrophobin fusion protein comprising the hydrophobin protein and one or more elements selected from the group consisting of signal sequence, fusion partner polypeptide, linker sequence, and purification sequence.

4. The method of claim 3 , wherein the hydrophobin fusion protein comprises a signal sequence.

5. The method of claim 4 , wherein the secretion signal sequence is encoded by:
an exogenous nucleic acid coding for a protein having an amino acid sequence with at least 80% identity to SEQ ID NO: 24.

6. The method of claim 3 , wherein the hydrophobin fusion protein comprises a fusion partner polypeptide and the fusion partner polypeptide is encoded by an exogenous nucleic acid coding for a protein having an amino acid sequence with at least 70% identity to SEQ ID NO: 16.

7. The method of claim 2 , wherein the promoter is a constitutive, pathogen-inducible promoter, a mesophyll-specific promoter or an epidermis specific-promoter.

8. A transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell having resistance to Phakopsora infection, comprising an exogenous nucleic acid coding for a hydrophobin protein having an amino acid sequence with at least 90% identity to SEQ ID NO: 42 in functional linkage with a promoter, wherein expression of the exogenous nucleic acid leads to increased resistance to Phakopsora infection thereto as compared to a wild type soybean plant, wild type soybean plant part, or wild type soybean plant cell.

9. A method for the production of a transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell having increased resistance to Phakopsora infection, comprising:
(a) introducing an expression cassette comprising:
an exogenous nucleic acid coding for a protein having an amino acid sequence with at least 90% identity to SEQ ID NO: 42 in functional linkage with a promoter; and
(b) generating a transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell from the plant, plant part or plant cell,
wherein expression of the exogenous nucleic acid leads to increased resistance to Phakopsora infection thereto as compared to a wild type soybean plant, wild type soybean plant part, or wild type soybean plant cell.

10. The method of claim 9 , further comprising the step of harvesting the seeds of the transgenic plant and planting the seeds and growing the seeds to plants, wherein the grown plants comprise:
the exogenous nucleic acid encoding a protein having an amino acid sequence with at least 90% identity to SEQ ID NO: 42.

11. A harvestable part of the transgenic plant of claim 8 , wherein the harvestable part of the transgenic plant comprises the exogenous hydrophobin nucleic acid encoding a protein having an amino acid sequence with at least 90% identity to SEQ ID NO: 42.

12. A product produced from the transgenic plant of claim 8 , wherein the product comprises the exogenous hydrophobin nucleic acid encoding a protein having an amino acid sequence with at least 90% identity to SEQ ID NO: 42.

13. A method for the production of a product comprising:
a) growing the transgenic plant of claim 8 ; and
b) producing said product from or by the plant or a part of the plant,
wherein the product comprises the exogenous hydrophobin nucleic acid encoding a protein having an amino acid sequence with at least 90% identity to SEQ ID NO: 42.

14. The method of claim 13 , wherein said product is produced from or by harvestable parts of the plant.

15. The method of claim 1 , wherein the fungal pathogen is Phakopsora meibomiae, Phakopsora pachyrhizi , or a combination thereof.

16. The method of claim 1 , wherein the plant is selected from the group consisting of beans, soya, pea, clover, kudzu, lucerne, lentils, lupins, vetches, groundnut, rice, wheat, barley, arabidopsis , lentil, banana, canola, cotton, potato, corn, sugar cane, alfalfa, and sugar beet.

17. A method for breeding a fungal resistant plant comprising:
(a) crossing the transgenic plant of claim 8 with a second plant;
(b) obtaining seed from the cross of step (a);
(c) planting said seeds and growing the seeds to plants; and
(d) selecting from the plants produced in step (c) plants expressing the hydrophobin protein.

18. The method of claim 1 , wherein exogenous nucleic acid encodes a hydrophobin protein with at least 95% identity to SEQ ID NO: 42 over the entire length of SEQ ID NO: 42.

19. The method of claim 1 , wherein exogenous nucleic acid encodes a hydrophobin protein with at least 98% identity to SEQ ID NO: 42 over the entire length of SEQ ID NO: 42.

20. The transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell of claim method of claim 8 , wherein exogenous nucleic acid encodes a hydrophobin protein comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 42.

21. The transgenic soybean plant, transgenic soybean plant part, or transgenic soybean plant cell of claim method of claim 8 , wherein exogenous nucleic acid encodes a hydrophobin protein comprising an amino acid sequence with at least 98% identity to SEQ ID NO: 42.

Assignees

• Basf Plant Science Co Gmbh

Inventors

• Schultheiss Holger
• Flachmann Ralf
• Subkowski Thomas
• Bollschweiler Claus

Classifications

• C07K14/38

  from Aspergillus · CPC title

• C12N15/625

  containing a sequence coding for a signal sequence · CPC title

• A01N37/18

  containing the group —CO—N<, e.g. carboxylic acid amides or imides; Thio analogues thereof · CPC title

• C07K14/37

  from fungi · CPC title

• C12N15/8282 Primary

  for fungal resistance · CPC title