Regulating alkaloids
Assignee22nd Century Limited, LLC
InventorsTakashi Hashimoto, Akira Kato
Life of this patent
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Abstract
MPO1 and MPO2 can be regulated for either decreasing or increasing alkaloid levels in plants, in particular in Nicotiana plants. In particular, suppressing or overexpressing one or more of MPO1 and MPO2 may be used to decrease or increase nicotine and nicotinic alkaloid levels in tobacco plants. Suppression or overexpression of one or more of MPO1 and MPO2 may be used in combination with modification of expression of other genes encoding enzymes on the nicotinic alkaloid biosynthetic pathway such as A622, NBB1, PMT, and QPT.
Claims13
1-36. (canceled)
A method of producing an N-methylputrescine oxidase-1 (MPO1) enzyme, comprising:
- (a) introducing into a host cell a heterologous nucleic acid construct comprising a nucleotide sequence selected from the group consisting of:
- (i) a nucleotide sequence set forth in SEQ ID NO:
- 1;
- (ii) a nucleotide sequence set forth in SEQ ID NO:
- 5;
- (iii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:
- 2;
- and
- (iv) any combination of (i)-(iii);
- and
- (b) growing the cell under conditions such that N-methylputrescine oxidase-1 (MPO1) is produced.
A method for increasing nicotine in a Nicotiana plant, comprising
- (a) introducing into the plant a vector comprising a nucleotide sequence selected from the group consisting of:
- (i) a nucleotide sequence set forth in SEQ ID NO:
- 1;
- (ii) a nucleotide sequence set forth in SEQ ID NO:
- 5;
- (iii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:
- 2;
- and
- (iv) any combination of (i)-(iii);
- and
- (b) growing the plant under conditions which allow for the expression of N-methylputrescine oxidase-1 (MPO1) from the nucleotide sequence;
- wherein expression of the MPO1 results in the plant having an increased nicotine content relative to a control plant.
A method for increasing the ratio of nicotine to total alkaloids in a tobacco plant, comprising:
- (a) introducing into the plant a nucleic acid construct comprising:
- (i) an expression cassette comprising at least one nucleotide sequence selected from the group consisting of a nucleotide sequence set forth in SEQ ID NO:
- 1, a nucleotide sequence set forth in SEQ ID NO:
- 5, and a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO:
- 2;
- (ii) a putrescine N-methyltransferase (PMT) expression cassette;
- and
- (iii) a quinolate phosphoribosyl transferase (QPT) suppression cassette;
- and
- (b) growing the plant under conditions which allow for overexpression of MPO1, overexpression of PMT, and suppression of QPT,
- wherein overexpression of MPO1, overexpression of PMT, and suppression of QPT results in the plant having an increased ratio of nicotine to total alkaloids in the plant as compared to a non-transformed control plant.
Dependencies are inferred from the claim text.