Gene conferring resistance to Cercospora beticola in beets

Patent metadata

Field	Value
Publication number	US-11597944-B2
Application number	US-202017005903-A
Country	US
Kind code	B2
Filing date	Aug 28, 2020
Priority date	Feb 18, 2019
Publication date	Mar 7, 2023
Grant date	Mar 7, 2023

Abstract

A more efficient breeding against Cercospora leaf spot disease, or the development of new resistant lines, is enabled via the provision of the Cercospora resistance-mediating gene according to the invention; in particular, a dominant resistance effect in the target plant is evoked by the property of the identified gene alone. The Cercospora resistance-mediating gene, and embodiments of the present invention that are described in the preceding, offer additional applications, e.g., the use of the resistant gene allele in cis-genetic or trans-genetic approaches, with the goal of developing new resistant cultivars.

First claim

What is claimed is:

1. A plant of the subspecies Beta vulgaris subsp. vulgaris comprising a nucleic acid molecule encoding a polypeptide that is able to confer resistance to Cercospora beticola in a plant in which the polypeptide is expressed, characterized in that the nucleic acid molecule comprises a nucleotide sequence which is selected from the group consisting of:
(a) a nucleotide sequence that encodes a polypeptide having the amino acid sequence according to SEQ ID NO: 3; and
(b) a nucleotide sequence that encodes a polypeptide which has an amino acid sequence that is at least 95% identical to the amino acid sequence according to SEQ ID NO: 3.

2. The plant according to claim 1 , wherein the plant is a hybrid plant.

3. The plant according to claim 1 , wherein the plant is a double haploid plant.

4. The plant according to claim 1 , wherein the resistance is dominant.

5. The plant according to claim 1 , wherein the plant comprises the nucleic acid molecule homozygous.

6. The plant according to claim 1 , wherein the plant comprises the nucleic acid molecule as an introgression.

7. The plant according to claim 1 , wherein the plant is a variety.

8. A storage organ of the plant of the subspecies Beta vulgaris subsp. vulgaris according to claim 1 .

9. A seed of the plant according to claim 1 , wherein the seed comprises the nucleic acid molecule.

10. A seed according to claim 9 , wherein the seed has been primed.

11. The hybrid plant according to claim 2 , wherein the plant has tolerance to at least one herbicide.

12. The hybrid plant according to claim 11 , wherein the at least one herbicide is selected from the group consisting of: glyphosate, ALS-inhibitor herbicide and glufosinate.

13. The plant according to claim 12 , wherein the tolerance to glyphosate is obtainable from deposition NCIMB 41158 or NCIMB 41159 and/or wherein the tolerance to an ALS-inhibitor herbicide is obtainable from deposition NCIMB 41705.

14. The plant according to claim 12 , wherein the tolerance to glyphosate is conferred by a DNA fragment selected from the list consisting of:
a) a DNA fragment of the genomic DNA of the plant or portions thereof which may be amplified via polymerase chain reaction with a first primer that has the nucleotide sequence of SEQ ID NO: 81, and a second primer that has the nucleotide sequence of SEQ ID NO: 82, wherein the DNA fragment is at least 95% identical to the nucleotide sequence of SEQ ID NO: 83, and/or
b) a DNA fragment of the genomic DNA of the plant or portions thereof which may be amplified via polymerase chain reaction with a first primer that has the nucleotide sequence of SEQ ID NO: 84, and a second primer that has the nucleotide sequence of SEQ ID NO: 85, wherein the DNA fragment is at least 95% identical to the nucleotide sequence of SEQ ID NO: 86, and/or
c) a DNA fragment of the genomic DNA of the plant or portions thereof which may be amplified via polymerase chain reaction with a first primer that has the nucleotide sequence of SEQ ID NO: 87, and a second primer that has the nucleotide sequence of SEQ ID NO: 88, wherein the DNA fragment is at least 95% identical to the nucleotide sequence of SEQ ID NO: 89.

15. The plant according to claim 12 , wherein the tolerance to an ALS-inhibitor herbicide is conferred by an acetolactate synthase gene encoding an acetolactate synthase protein which, as a result of a mutation, has at position 569 a different amino acid than tryptophan.

16. The plant according to claim 12 , wherein the mutated acetolactate synthase gene comprises the sequence according to SEQ ID No. 91.

17. A method for identifying a Cercospora beticola -resistant plant of the subspecies Beta vulgaris subsp. vulgaris according to claim 14 , the method comprising:
(i) detection of the presence and/or expression of the nucleic acid molecule in the plant or a portion of the plant; and/or
(ii) detection of at least one marker locus in the nucleotide sequence of the nucleic acid molecule or in a cosegregating region thereof; and
(iii) selection of the Cercospora beticola -resistant plant,
wherein step (i) and/or step (ii) use sequence-specific amplification and/or detection of amplified variable sequences of the plant genome.

18. The method according to claim 17 , wherein the detection in step (i) or (ii) involves oligonucleotides linked to a fluorescent dye in order to generate a fluorescence signal.

19. The method according to claim 17 , wherein the fluorescent dye is selected from the group consisting of: FAM, AlexaFluor, ATTO, Dabcyl, HEX, Rox, TET, Texas Red, and Yakima Yellow.

20. A method for farming of plants of the subspecies Beta vulgaris subsp. vulgaris , including
(i) the planting of a seed according to claim 9 , and
(ii) cultivation of the plants from (i) or descendants thereof,
wherein the method counteracts an infestation of the cultivated plants with Cercospora.

21. A method for producing Cercospora resistant plants of the subspecies Beta vulgaris subsp. vulgaris according to claim 1 comprising:
(i) introducing the nucleic acid molecule into the plant;
(ii) expressing the polypeptide that is able to confer resistance to Cercospora beticola ; and
(iii) cultivating the plant of step (i).

22. A method of producing an agronomical sugar beet plant of the genus Beta that displays improved resistance to Cercospora beticola , the method comprising introgressing into said plant a chromosomal interval that confers resistance to Cercospora beticola , wherein the chromosomal interval maps to a position between a sequence represented by a marker selected from the group consisting of: s4p42932s01 and s4p4295s01 and a sequence represented by a marker selected from the group consisting of: s4p4301s01 and sxh0678s01,
wherein the chromosomal interval comprises a nucleotide sequence encoding a polypeptide that is able to confer resistance to Cercospora beticola in a plant in which the polypeptide is expressed wherein the nucleotide sequence is selected from the group consisting of:
(a) a nucleotide sequence that encodes a polypeptide having the amino acid sequence according to SEQ ID NO: 3; and
(b) a nucleotide sequence that encodes a polypeptide which has an amino acid sequence that is at least 95% identical to the amino acid sequence according to SEQ ID NO: 3.

Assignees

• Kws Saat Se & Co Kgaa

Inventors

• Törjék Otto
• Borchardt Dietrich
• Rekoske Margaret
• Mechelke Wolfgang
• Schulz Britta
• Lein Jens Christoph

Classifications

• C12N15/8282 Primary

  for fungal resistance · CPC title

• A01H6/024

  Beta vulgaris [beet] · CPC title

• C12N15/8213

  Targeted insertion of genes into the plant genome by homologous recombination · CPC title

• C07K14/415

  from plants · CPC title

• A01H5/06

  Roots · CPC title