US 11,778,965 B2
Alteration of tobacco alkaloid content through modification of specific cytochrome P450 genes
Ralph E. Dewey, Apex, NC (US); Balazs Siminszky, Neuchatel (CH); Steven W. Bowen, Raleigh, NC (US); and Lily Gavilano, Raleigh, NC (US)
Assigned to North Carolina State University, Raleigh, NC (US); and University of Kentucky Research Foundation, Lexington, KY (US)
Filed by North Carolina State University, Raleigh, NC (US); and University of Kentucky Research Foundation, Lexington, KY (US)
Filed on Sep. 22, 2021, as Appl. No. 17/481,799.
Application 17/481,799 is a continuation of application No. 16/511,596, filed on Jul. 15, 2019, granted, now 11,140,843.
Application 16/511,596 is a continuation of application No. 15/883,931, filed on Jan. 30, 2018, granted, now 10,383,299, issued on Aug. 20, 2019.
Application 15/883,931 is a continuation of application No. 14/878,083, filed on Oct. 8, 2015, granted, now 9,913,451, issued on Mar. 13, 2018.
Application 14/878,083 is a continuation of application No. 12/971,801, filed on Dec. 17, 2010, granted, now 9,187,759, issued on Nov. 27, 2015.
Application 12/971,801 is a continuation of application No. 11/580,765, filed on Oct. 13, 2006, granted, now 7,884,263, issued on Feb. 8, 2011.
Application 11/580,765 is a continuation in part of application No. PCT/US2005/005665, filed on Feb. 23, 2005.
Prior Publication US 2022/0000055 A1, Jan. 6, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. A01H 5/12 (2018.01); A01H 6/82 (2018.01); C12N 9/02 (2006.01); C12N 15/82 (2006.01); A24B 13/00 (2006.01); A24B 15/10 (2006.01); C12Q 1/6895 (2018.01)
CPC A01H 5/12 (2013.01) [A01H 6/823 (2018.05); A24B 13/00 (2013.01); A24B 15/10 (2013.01); C12N 9/0073 (2013.01); C12N 9/0077 (2013.01); C12N 15/8243 (2013.01); C12Q 1/6895 (2013.01); C12Q 2600/13 (2013.01); C12Q 2600/156 (2013.01)] 16 Claims
 
1. A method for reducing the level of nornicotine or N′-nitrosonornicotine in a Nicotiana tabacum plant, the method comprising generating a stable mutation in a nicotine demethylase in the Nicotiana tabacum plant, wherein the stable mutation inhibits the expression or function of the nicotine demethylase, wherein the nicotine demethylase is a polypeptide comprising an amino acid sequence at least 98% identical to SEQ ID NO: 4, and wherein the amount of nornicotine or N′-nitrosonornicotine is reduced by greater than about 10% compared to a control Nicotiana tabacum plant comprising a wild-type allele of the nicotine demethylase.