Amino polyol amine oxidase polynucleotides and related polypeptides and methods of use

A recombinant expression cassette comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• c) a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• d) a polynucleotide complementary to a polynucleotide of (a) through (c).

A vector comprising a recombinant expression cassette comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• c) a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• d) a polynucleotide complementary to a polynucleotide of (a) through (c).

A host cell comprising a recombinant expression cassette comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• c) a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• d) a polynucleotide complementary to a polynucleotide of (a) through (c).

A transformed plant comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• c) a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• d) a polynucleotide complementary to a polynucleotide of (a) through (c).

A plant seed comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• c) a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• d) a polynucleotide complementary to a polynucleotide of (a) through (c).

A method of reducing pathogenicity of a fungus producing fumonisin or a structurally related mycotoxin, comprising:

• a) transforming a plant cell with a vector comprising a polynucleotide selected from the group consisting of:
• i. a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• ii. a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• iii. a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• iv. a polynucleotide complementary to a polynucleotide of i. through iii. operably linked to a promoter;
• b) growing the plant cell under plant growing conditions;
• and
• c) inducing expression of said polynucleotides for a time sufficient for amounts of
• the fumonisin esterase and APAO enzymes to accumulate to levels that can inhibit the fungus.

A method of making an APAO enzyme comprising the steps of:

• a) expressing a polynucleotide in a recombinantly engineered cell, wherein the polynucleotide is selected from the group consisting of:
• i. a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• ii. a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• iii. a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• iv. a polynucleotide complementary to a polynucleotide of i. through iii. operably linked to a promoter;
• and
• b) purifying the enzyme.

A method of making an APAO enzyme comprising the steps of:

• a) expressing a polynucleotide in a plant, wherein said polynucleotide is selected from the group consisting of:
• i. a polynucleotide comprising at least 20 contiguous bases selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• ii. a polynucleotide comprising at least 70% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• iii. a polynucleotide comprising at least 80% sequence identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• iv. a polynucleotide complementary to a polynucleotide of i. through iii. operably linked to a promoter;
• and
• b) purifying the enzyme from the plant seed or other plant parts.

An isolated polynucleotide comprising a polynucleotide selected from:

• a) a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• c) a polynucleotide complementary to a polynucleotide of (a) through (b).

A recombinant expression cassette comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• c) a polynucleotide complementary to a polynucleotide of (a) through (b).

A vector comprising a recombinant expression cassette comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• c) a polynucleotide complementary to a polynucleotide of (a) through (b).

A host cell comprising a recombinant expression cassette comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• c) a polynucleotide complementary to a polynucleotide of (a) through (b).

A transformed plant comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• c) a polynucleotide complementary to a polynucleotide of (a) through (b).

A plant seed comprising a polynucleotide selected from the group consisting of:

• a) a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• b) a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• c) a polynucleotide complementary to a polynucleotide of (a) through (b).

A method of reducing pathogenicity of a fungus producing fumonisin or a structurally related mycotoxin, comprising:

• a) transforming a plant cell with a vector comprising a polynucleotide selected from the group consisting of:
• i. a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• ii. a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• iii. a polynucleotide complementary to a polynucleotide of (a) through (b).
• b) growing the plant cell under plant growing conditions;
• and
• c) inducing expression of said polynucleotides for a time sufficient for amounts of the fumonisin esterase and APAO enzymes to accumulate to levels that can inhibit the fungus.

A method of making an APAO enzyme comprising the steps of:

• a) expressing a polynucleotide in a recombinantly engineered cell, wherein the polynucleotide is selected from the group consisting of:
• i. a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• ii. a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• iii. a polynucleotide complementary to a polynucleotide of (a) through (b). and
• b) purifying the enzyme.

A method of making an APAO enzyme comprising the steps of:

• a) expressing a polynucleotide in a plant, wherein said polynucleotide is selected from the group consisting of:
• i. a polynucleotide which hybridizes under high stringency conditions to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• ii. a polynucleotide comprising at least 90% identity to a polynucleotide selected from SEQ ID NO:
• 5, SEQ ID NO:
• 10, and SEQ ID NO:
• 22;
• and
• iii. a polynucleotide complementary to a polynucleotide of (a) through (b). and
• b) purifying the enzyme from the plant seed or other plant parts.