US 12,084,670 B2
Regulating alkaloids
Takashi Hashimoto, Ikoma (JP); and Akira Kato, Nara (JP)
Assigned to 22nd Century Limited, LLC, Buffalo, NY (US)
Filed by 22nd Century Limited, LLC, Buffalo, NY (US)
Filed on Jan. 5, 2023, as Appl. No. 18/093,784.
Application 18/093,784 is a division of application No. 16/925,069, filed on Jul. 9, 2020, granted, now 11,572,568.
Application 16/003,066 is a division of application No. 14/822,105, filed on Aug. 10, 2015, granted, now 9,994,860.
Application 14/822,105 is a division of application No. 11/941,950, filed on Nov. 18, 2007, granted, now 9,102,948.
Application 16/925,069 is a continuation of application No. 16/003,066, filed on Jun. 7, 2018, granted, now 10,731,172.
Claims priority of provisional application 60/866,352, filed on Nov. 17, 2006.
Prior Publication US 2023/0340510 A1, Oct. 26, 2023
Int. Cl. C12N 15/82 (2006.01); A24B 13/00 (2006.01); A24B 15/10 (2006.01); A61K 31/455 (2006.01); C12N 9/02 (2006.01); C12N 9/06 (2006.01)
CPC C12N 15/8218 (2013.01) [A24B 13/00 (2013.01); A24B 15/10 (2013.01); A61K 31/455 (2013.01); C12N 9/0004 (2013.01); C12N 9/0012 (2013.01); C12N 9/0022 (2013.01); C12N 15/8216 (2013.01); C12N 15/8243 (2013.01); C12N 15/8279 (2013.01); C12N 15/8286 (2013.01); C12Y 104/03006 (2013.01); Y02A 40/146 (2018.01)] 4 Claims
 
1. 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.