# US 11,299,743 B2 · Genetic locus imparting a low anatabine trait in tobacco and methods of using > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,299,743 B2 - Document ID: US11299743B2 - Status: Granted - Kind: B2 - Application number: 16/825,394 - Filed: 2020-03-20 - Published: 2020-09-03 - Granted: 2022-04-12 - Earliest claimed: 2014-07-08 - Assignees: Altria Client Services LLC - Applicants: Altria Client Services LLC - Inventors: Chengalrayan Kudithipudi; Alec J. Hayes; Robert Frank Hart, III; Yanxin Shen; Jesse Frederick; Dongmei Xu - Web page: https://patentgrep.com/patent/US11299743B2 ## CPC - C12N 15/8261: {with agronomic (input) traits, e.g. crop yield} - A01H 1/045: {using molecular markers} - A01H 1/06: Processes for producing mutations, e.g. treatment with chemicals or with radiation (specific mutations prepared by genetic engineering on plant cell or plant tissues C12N15/00 {; process for producing transgenic plants C12N15/82}) - A01H 5/12: Leaves - A01H 6/823: {Nicotiana, e.g. tobacco} - A24B 13/00: Tobacco for pipes, for cigars, e.g. cigar inserts, or for cigarettes; Chewing tobacco; Snuff (reconstituted tobacco products A24B3/14) - C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]} - C12N 15/8243: {involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine} - C12Q 1/6895: for plants, fungi or algae - C12Q 2600/13: Plant traits - C12Q 2600/156: Polymorphic or mutational markers - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES - A24B: MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract Provided herein are genetic markers and a coding sequence associated with a low- or ultra-low anatabine trait in tobacco. ## Claims ### Claim 1 A tobacco plant having an induced mutation in an endogenous gene, wherein the endogenous gene has a sequence 100% identical to SEQ ID NO: 7. ### Claim 2 (depends on 1) The tobacco plant of claim 1, wherein the mutant plant exhibits reduced expression or activity of quinolinate synthase as compared to a control plant lacking the induced mutation. ### Claim 3 (depends on 1) The tobacco plant of claim 1, wherein leaf from the mutant plant comprises a reduced amount of at least one alkaloid as compared to leaf from a control plant lacking the induced mutation. ### Claim 4 (depends on 3) The tobacco plant of claim 3, wherein the alkaloid is anatabine. ### Claim 5 (depends on 1) The tobacco plant of claim 1, wherein cured leaf from the mutant plant exhibits a reduced amount of at least one tobacco-specific nitrosamine (TSNA) as compared to cured leaf from a control tobacco plant lacking the induced mutation. ### Claim 6 (depends on 5) The tobacco plant of claim 5, wherein the at least one TSNA is N′-nitrosoanatabine (NAT). ### Claim 7 (depends on 1) Seed produced by the tobacco plant of claim 1, wherein the seed comprises the induced mutation. ### Claim 8 (depends on 1) The tobacco plant of claim 1, wherein the induced mutation provides an amino acid substitution at a position corresponding to the cysteine residue at position 487 or the valine residue at position 516 of SEQ ID NO: 8. ### Claim 9 (depends on 1) The tobacco plant of claim 1, wherein the induced mutation comprises a tyrosine at amino acid position 487 of SEQ ID NO: 8. ### Claim 10 (depends on 1) The tobacco plant of claim 1, wherein the induced mutation comprises an isoleucine at position 516 of SEQ ID NO: 8. ### Claim 11 (depends on 4) The tobacco plant of claim 4, wherein the reduced amount of anatabine comprises a reduction of about 50% to about 95% as compared to a control plant lacking the induced mutation. ### Claim 12 (depends on 1) The tobacco plant of claim 1, wherein cured leaf from the tobacco plant comprises an anatabine percentage of less than 0.5%. ### Claim 13 (depends on 1) The tobacco plant of claim 1, wherein the induced mutation is in a residue fully conserved in the wild-type quinolinate synthase proteins. ### Claim 14 A method for producing a tobacco seed comprising: (a) crossing a first tobacco plant having an induced mutation in an endogenous gene, wherein the endogenous gene has a sequence 100% identical to SEQ ID NO: 7 with a second tobacco plant to produce at least one progeny seed; and (b) selecting at least one progeny seed from step (a) that comprises said induced mutation. ### Claim 15 (depends on 14) The method of claim 14, wherein the first tobacco plant exhibits reduced expression or activity of quinolinate synthase as compared to a control plant lacking the induced mutation. ### Claim 16 (depends on 14) The method of claim 14, wherein the induced mutation provides an amino acid substitution at a position corresponding to the cysteine residue at position 487 or the valine residue at position 516 of SEQ ID NO: 8. ### Claim 17 A method of making a tobacco plant comprising: (a) inducing mutagenesis in a Nicotiana tabacum cell to produce a mutagenized cell; (b) obtaining one or more plants regenerated from the mutagenized cell of step (a); and (c) identifying at least one plant from step (b) that comprises an induced mutation in an endogenous gene, wherein the endogenous gene has a sequence 100% identical to SEQ ID NO: 7. ### Claim 18 (depends on 17) The method of claim 17, wherein the at least one plant identified in step (c) exhibits reduced expression or activity of quinolinate synthase as compared to a control plant lacking the induced mutation.