US 2003/0154513 A1 · Methyltransferase genes and uses thereof

Abstract

The present invention relates to genes associated with the tocopherol biosynthesis pathway. More particularly, the present invention provides and includes nucleic acid molecules, proteins, and antibodies associated with genes that encode polypeptides that have methyltransferase activity. The present invention also provides methods for utilizing such agents, for example in gene isolation, gene analysis and the production of transgenic plants. Moreover, the present invention includes transgenic plants modified to express the aforementioned polypeptides. In addition, the present invention includes methods for the production of products from the tocopherol biosynthesis pathway.

Claims97

Dependent claims

A substantially purified nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 50, and 85.

A substantially purified nucleic acid molecule comprising a nucleic acid sequence that encodes a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-38.

A substantially purified nucleic acid molecule comprising as operably linked components:

(A) a promoter region which functions in a plant cell to cause the production of an mRNA molecule;
(B) a heterologous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:
19-31 and 33-38.

A substantially purified protein comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-38.

An antibody capable of specifically binding a substantially purified protein with an amino acid sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-38.

A transformed plant comprising an exogenous nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 50, and 85 and complements thereof.

A transformed soybean line A3244 plant, comprising an exogenous nucleic acid molecule comprising a nucleic acid sequence of SEQ ID NO:

2, operably linked to a nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of p7S, SEQ ID NO:
81, and SEQ ID NO:
82.

A transformed plant having an exogenous nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 50, and 85 and complements thereof, wherein said transformed plant produces a seed with increased α-tocotrienol levels relative to a plant with a similar genetic background but lacking said exogenous nucleic acid molecule.

A transformed plant having an exogenous nucleic acid molecule that encodes a polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-40.

A transformed plant having an exogenous nucleic acid molecule that encodes a polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-40, wherein said transformed plant produces a seed with increased α-tocotrienol levels relative to a plant with a similar genetic background but lacking said exogenous nucleic acid molecule.

A transformed plant having an exogenous nucleic acid molecule that encodes a polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

46-49.

A method for reducing expression of MT1 or GMT in a plant comprising:

(A) transforming a plant with a nucleic acid molecule, said nucleic acid molecule having an exogenous promoter region which functions in plant cells to cause the production of a mRNA molecule, wherein said exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein said transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of:
SEQ ID NOs:
2-17, 42-45, 50, and 85;
and wherein said transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA sequence;
and (B) growing said transformed plant.

2 more elements

A transformed plant comprising a nucleic acid molecule comprising an exogenous promoter region which functions in plant cells to cause the production of a mRNA molecule, wherein said exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein said transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 42-45, 50, and 85, and wherein said transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA sequence.

A method for increasing the γ-tocopherol content in a plant comprising:

(A) transforming a plant with a nucleic acid molecule, said nucleic acid molecule having an exogenous promoter region which functions in plant cells to cause the production of a mRNA molecule, wherein said exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein said transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
2-17, 50, and 85;
and wherein said transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA sequence;
and (B) growing said transformed plant.

1 more element

A transformed plant comprising:

(A) a first nucleic acid molecule comprising an exogenous promoter region which functions in plant cells to cause the production of a mRNA molecule, wherein said exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein said transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
2-17, 50, and 85, and wherein said transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA sequence;
and (B) a second nucleic acid molecule comprising an exogenous promoter region which functions in plant cells to cause the production of a mRNA molecule, wherein said exogenous promoter region is linked to a nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NOs:
42-45.

1 more element

A method of producing a plant having a seed with an increased α-tocopherol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
2-17, 50, and 85;
and (B) growing said transformed plant.

A method of producing a plant having a seed with an increased α-tocotrienol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
2-17, 50, and 85;
and (B) growing said transformed plant.

A method of producing a plant having a seed with an increased α-tocopherol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a sequence encoding an polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:
19-31 and 33-40;
and (B) growing said transformed plant.

A method of producing a plant having a seed with an increased α-tocotrienol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a sequence encoding an polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:
19-31 and 33-40;
and (B) growing said transformed plant.

A method of producing a plant having a seed with an increased γ-tocopherol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleic acid sequence that encodes an polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:
46-49 and (B) growing said transformed plant.

A method of producing a plant having a seed with an increased γ-tocopherol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
42-45 and (B) growing said transformed plant.

A method of producing a plant having a seed with an increased γ-tocotrienol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleic acid sequence that encodes a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NO:
47 through SEQ ID NO:
49;
and (B) growing said transformed plant.

1 more element

A method of producing a plant having a seed with an increased γ-tocotrienol level comprising:

(A) transforming said plant with a nucleic acid molecule, wherein said nucleic acid molecule comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
42-46;
and (B) growing said transformed plant.

A seed derived from a transformed plant having an exogenous nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 50, and 85, wherein said seed has an increased α-tocopherol level relative to a seed from a plant having a similar genetic background but lacking said exogenous nucleic acid molecule.

A seed derived from a transformed plant having an exogenous nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 50, and 85, wherein said seed has an increased α-tocotrienol level relative to a seed from a plant having a similar genetic background but lacking said exogenous nucleic acid molecule.

Oil derived from a seed of a transformed plant, wherein said transformed plant comprises an exogenous nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

2-17, 42-45, 50, and 85.

Feedstock comprising a transformed plant or part thereof, wherein said transformed plant has an exogenous nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 42-45, 50, and 85.

A meal comprising plant material manufactured from a transformed plant, wherein said transformed plant has an exogenous nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 42-45, 50, and 85.

A seed derived from a transformed plant comprising an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-40 wherein said seed has an increased α-tocopherol level relative to seeds from a plant having a similar genetic background but lacking said exogenous nucleic acid molecule.

A seed derived from a transformed plant comprising an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-40 wherein said seed has an increased α-tocotrienol level relative to seeds from a plant having a similar genetic background but lacking said exogenous nucleic acid molecule.

Oil derived from a seed of a transformed plant, wherein said transformed plant comprises an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

19-31, 33-40, and 46-49.

Feedstock comprising a transformed plant or part thereof, wherein said transformed plant comprises an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

19-31 and 33-40.

A meal comprising plant material manufactured from a transformed plant, wherein said transformed plant comprises an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

19-31, 33-40, and 46-49.

A seed derived from a transformed plant comprising an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

46-49, wherein said seed has an increased γ tocopherol level relative to seeds from a plant having a similar genetic background but lacking said exogenous nucleic acid molecule.

A seed derived from a transformed plant having an exogenous nucleic acid molecule encoding a polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NOs:

46-49, wherein said seed has an increased γ tocotrienol level relative to seeds from a plant having a similar genetic background but lacking said exogenous nucleic acid molecule.

46-49.

Meal comprising plant material manufactured from a transformed plant, wherein said transformed plant has an exogenous nucleic acid molecule encoding an amino acid sequence selected from the group consisting of SEQ ID NOs:

46-49.

A host cell comprising a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:

2-17, 42-45, 50, and 85 and complements thereof.

A transformed plant comprising an introduced first nucleic acid molecule that encodes a polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

19-31, 33-38, and 39-41, and an introduced second nucleic acid molecule encoding an enzyme selected from the group consisting of tyrA, slr1736, ATPT2, dxs, dxr, GGPPS, HPPD, GMT, MT1, tMT2, AANT1, slr 1737, and an antisense construct for homogentisic acid dioxygenase.

A transformed plant comprising an introduced first nucleic acid molecule that encodes a polypeptide molecule comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

46-49, and an introduced second nucleic acid molecule encoding an enzyme selected from the group consisting of tyrA, slr1736, ATPT2, dxs, dxr, GGPPS, HPPD, GMT, MT1, tMT2, AANT1, slr 1737, and an antisense construct for homogentisic acid dioxygenase.

A plant comprising an introduced nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of 42-45, wherein said transformed plant produces a seed having increased total tocopherol relative to seed of a plant with a similar genetic background but lacking said introduced nucleic acid molecule.

A plant comprising an introduced nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of 2-17, 50, 85, wherein said transformed plant produces a seed having increased total tocopherol relative to seed of a plant with a similar genetic background but lacking said introduced nucleic acid molecule.

A plant comprising a first introduced nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of 2-17, 50, and 85 and a second introduced nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of 42-45, wherein said transformed plant produces a seed having increased total tocopherol relative to seed of a plant with a similar genetic background but lacking both said introduced first nucleic acid molecule and said introduced second nucleic acid molecule.

Dependencies are inferred from the claim text.

Methyltransferase genes and uses thereof

Life of this patent

Methyltransferase genes and uses thereof

Life of this patent

Abstract

Claims97