Compositions and methods for fumonisin detoxification
AssigneePioneer Hi-Bred International, Inc.
InventorsJon Duvick, Joyce Maddox, Jacob Gilliam, Otto Folkerts, Oswald R. Crasta
Life of this patent
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Abstract
Compositions and methods for the complete detoxification of fumonisin and fumonisin degradation products are provided. Particularly, nucleotide sequences corresponding to the detoxification enzymes are provided. The sequences find use in preparing expression cassettes for the transformation of a broad variety of host cells and organisms.
Claims46
A host cell having stably incorporated at least one expression cassette comprising a promoter active in said cell operably linked to a nucleotide sequence, wherein said nucleotide sequence is heterologous to the cell and said nucleotide sequence is selected from the group consisting of the nucleotide sequence set forth in SEQ ID NOS:
- 1, 2, 4, 6, 7, 9, and 10.
A plant having stably incorporated within its genome at least one nucleic acid molecule operably linked to a promoter active in said plant, said molecule comprising the nucleotide sequence set forth in any one of SEQ ID NO:2, 4, 7, or 10.
A method of reducing pathogenicity of a fungus producing fumonisin, comprising:
- a) stably integrating into the genome of a plant cell an expression cassette comprising at least one nucleotide sequence operably linked to a promoter active in said cell, wherein said nucleotide sequence is set forth in any one of SEQ ID NO:
- 1, 2, 4, 6, 7, 9, or 10;
- b) growing the plant cell under the growing conditions;
- and
- c) inducing expression of said nucleotide sequence for a time sufficient for amounts of the degradative enzyme or transporter to accumulate to levels that can reduce pathogenicity of said fungus.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:1.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:2.
An isolated nucleic acid molecule encoding the amino acid sequence set forth in SEQ ID NO:3.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:4.
An isolated nucleotide molecule encoding the amino acid sequence set forth in SEQ ID NO:5.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:6.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:7.
An isolated nucleic acid molecule encoding the amino acid sequence set forth in SEQ ID NO:8.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:9.
An isolated nucleic acid molecule comprising a nucleotide sequence set forth in SEQ ID NO:10.
An isolated nucleic acid molecule encoding the amino acid sequence set forth in SEQ ID NO:11.
An expression cassette comprising the nucleotide sequence of any one of SEQ ID NOS:
- 1, 4, 6, 9, or 10.
An expression cassette comprising the nucleotide sequence of any one of SEQ ID NOS:
- 2, 4, 7, or 10.
An isolated nucleic acid molecule comprising a nucleotide sequence having at least 80% identity to the nucleotide sequence of SEQ ID NO:2, wherein said sequence encodes a polypeptide having monooxygenase activity.
An isolated nucleic acid molecule selected from the group consisting of a nucleic acid sequence having at least 95% identity to the nucleotide sequence of any one of SEQ ID NOS:
- 2, 4, 7, or 10 wherein said sequence encodes a polypeptide having fumonisin detoxification activity.
An isolated nucleic acid molecule comprising a nucleotide sequence having at least 80% identity to the nucleotide sequence of SEQ ID NO:7, wherein said sequence encodes a polypeptide having permease activity.
A host cell having stably incorporated at least one expression cassette comprising a promoter active in said cell operably linked to a nucleotide sequence, wherein said nucleotide sequence is heterologous to the cell and wherein:
- a) said nucleotide sequence is selected from the group consisting of:
- (i) a nucleotide sequence encoding the amino acid sequence set forth in one of SEQ ID NOS:3, 5, 8 or 11;
- (ii) a nucleotide sequence having at least 95% identity to the nucleotide sequence set forth in SEQ ID NOS:1, 2, 4, 6, 7, 9 or 10;
- (iii) a nucleotide sequence having at least 90% identity to the nucleotide sequence set forth in SEQ ID NOS:1, 2, 4, 6, 7, 9, or 10;
- (iv) a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence set forth in SEQ ID NOS:
- 1, 2, 4, 6, 7, 9, or 10;
- and,
- b) expression of said sequences reduces the pathogenicity of a fungus producing fumonisin.
A host cell having stably incorporated at least one expression cassette comprising a nucleotide sequence wherein:
- a) said nucleotide sequence is operably linked to a promoter that is active in said cell;
- b) said promoter is heterologous to the nucleotide sequence;
- and,
- c) said sequence is selected from the group consisting of a nucleotide sequence set forth in SEQ ID NOS:1, 2, 4, 6, 7, 9, or 10.
A host cell having stably incorporated at least one expression cassette wherein:
- a) said nucleotide sequence is operably linked to a promoter that is active in said cell;
- b) said promoter is heterologous to the nucleotide sequence;
- c) said sequence is selected from the group consisting of:
- (i) a nucleotide sequence encoding the amino acid sequence set forth in one of SEQ ID NOS:3, 5, 8 or 11;
- (ii) a nucleotide sequence having at least 95% identity to the nucleotide sequence set forth in SEQ ID NOS:1, 2, 4, 6, 7, 9, or 10;
- (iii) a nucleotide sequence having at least 90% identity to the nucleotide sequence set forth in SEQ ID NOS:1, 2, 4, 6, 7, 9, or 10;
- and,
- (iv) a nucleotide sequence having at least 80% identity to the nucleotide sequence set forth in SEQ ID NOS:1, 2, 4, 6, 7, 9, or 10;
- and,
- d) expression of said sequences reduces the pathogenicity of a fungus producing fumonisin.
Dependencies are inferred from the claim text.