IP Library › Granted Patent US 12,195,738
Granted Patent B2
US 12,195,738 · App. 17/597,249 · Granted Jan 14, 2025

Method for modifying alkaloid content in plants

Inventors: Sara Ben Khaled (London, GB); Francisco Anastacio De Abreu E Lima (London, GB)
Assignee: BRITISH AMERICAN TOBACCO (INVESTMENTS) LIMITED
C12N15/8243C07K14/415C12N15/8218
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,195,738
App. No.
17/597,249
Granted
Jan 14, 2025
Kind
B2
Abstract

The present invention relates to a method of modulating the alkaloid content of a plant or a part thereof, the method comprising modifying the plant by modulating the activity or expression of at least one gene encoding an RNA binding protein. The present invention also relates to a method of reducing the content of at least one tobacco specific nitrosamine (TSNA) precursor in tobacco, the method comprising modulating the activity or expression of at least one gene encoding an RNA binding protein.

Claims (24)

1. A method of decreasing or increasing the nicotine nornicotine, pseudoxynicotine (PON), anabasine and/or anatabine content of a tobacco plant or part thereof or tobacco cell or tobacco cell culture, wherein:

(A): the method of decreasing the nicotine, nornicotine, PON, anabasine and/or anatabine content comprises modifying said plant or part thereof or cell by decreasing the activity or expression of a gene encoding an RNA binding protein; and

(B): the method of increasing the nicotine, nornicotine, PON, anabasine and/or anatabine content comprises modifying said plant or part thereof or cell by increasing the activity or expression of a gene encoding an RNA binding protein,

wherein the RNA binding protein has an amino acid sequence comprising SEQ ID NO: 1 or a sequence which has at least 95% identity to SEQ ID NO: 1.

2. The method according to claim 1 , wherein the nicotine, nornicotine, PON, anabasine and/or anatabine content is reduced in comparison to a plant or part thereof or cell or cell culture or plant propagation material or leaf which has not been modified to decrease the activity or expression of the gene encoding said RNA binding protein having an amino acid sequence comprising SEQ ID NO: 1 or a sequence which has at least 95% identity to SEQ ID NO: 1.

3. A Nicotiana plant or part thereof or a Nicotiana cell or cell culture which has been modified to achieve a decrease or increase in nicotine, nornicotine, pseudoxynicotine (PON), anabasine and/or anatabine content in comparison to an unmodified plant or part thereof or unmodified cell culture, wherein:

(A): the modification to decrease the nicotine, nornicotine, PON, anabasine and/or anatabine content reduces the activity or expression of a gene encoding an RNA binding protein; and

(B): the modification to increase the nicotine, nornicotine, PON, anabasine and/or anatabine content increases the activity or expression of a gene encoding an RNA binding protein,

wherein the RNA binding protein has an amino acid sequence comprising SEQ ID NO: 1 or a sequence which has at least 95% identity to SEQ ID NO: 1.

4. A Nicotiana plant propagation material obtained from the Nicotiana plant or part thereof or the Nicotiana cell or cell culture according to claim 3 , wherein the plant propagation material is heterozygous or homozygous for the modification.

5. The Nicotiana plant or part thereof or the Nicotiana cell or cell culture according to claim 3 , or a Nicotiana plant propagation material obtained therefrom which is heterozygous or homozygous for the modification, wherein the nornicotine content is decreased.

6. A harvested leaf or a cut harvested leaf of the plant according to claim 3 , or a plant propagated from a plant propagation material obtained therefrom, wherein the plant propagated from the propagation material is heterozygous or homozygous for the modification.

7. A processed leaf obtained by processing the plant according to claim 3 , or a plant propagated from a plant propagation material obtained therefrom, wherein the plant propagated from the propagation material is heterozygous or homozygous for the modification.

8. The processed leaf according to claim 7 , wherein the leaf is processed by curing, fermenting, pasteurising or a combination thereof.

9. The processed leaf according to claim 7 , wherein the content of one or more TSNAs selected from N′-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N′-nitrosoanatabine (NAT) and N-nitrosoanabasine (NAB) is reduced.

10. The processed leaf according to claim 7 , wherein the processed leaf is a cut processed leaf.

11. A cured tobacco material made from the plant or part thereof according to claim 3 , or a plant propagated from a plant propagation material obtained therefrom, wherein the plant propagated from the propagation material is heterozygous or homozygous for the modification.

12. A tobacco blend comprising said cured tobacco material of claim 11 .

13. A mutant of a Nicotiana plant carrying a heritable mutation in a nucleotide sequence of a gene encoding an RNA binding protein wherein the gene is selected from SEQ ID NO: 2 or 3 or a sequence which has at least 95% identity to SEQ ID NO: 2 or 3, wherein said heritable mutation decreases the activity or expression of the gene encoding an RNA binding protein and wherein the mutant plant has decreased nicotine, nornicotine, PON, anabasine and/or anatabine relative to a comparable plant which does not carry said heritable mutation.

14. Progeny or seed of the mutant plant which carries the heritable mutation according to claim 13 .

15. A harvested leaf, a processed leaf or cured tobacco material produced from the plant according to claim 13 , wherein the harvested leaf, processed leaf or cured tobacco material carries the heritable mutation.

16. The processed leaf according to claim 7 , wherein the leaf is a flue-cured, air-cured, fire-cured, or sun-cured processed tobacco leaf.

17. The processed leaf according to claim 9 , wherein the content of NNN and/or NNK is reduced.

18. The processed leaf according to claim 9 , wherein the content of NNN is reduced.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 30, 2021
From: BEN KHALED, SARA; ANASTACIO DE ABREU E LIMA, FRANCISCO
To: BRITISH AMERICAN TOBACCO (INVESTMENTS) LIMITED
Reel/Frame 058506/0163 →
Priority Claims (1)
GB 1909563 · Jul 3, 2019 · national
Continuity (1)
Related Publication 20220340921A1 · Oct 27, 2022
References Cited (44)
US 6376750B1 · Yu et al. · 2002 [cited by applicant]
US 7798153B2 · Lawrence, Jr. · 2010 [cited by examiner]
US 8455719B2 · Frankard et al. · 2013 [cited by applicant]
US 9290773B2 · Edgerton · 2016 [cited by applicant]
US 9878004B2 · Williams et al. · 2018 [cited by applicant]
US 9879275B2 · Nadzan · 2018 [cited by examiner]
US 10138492B2 · Nadzan et al. · 2018 [cited by applicant]
US 10167482B2 · Coffin · 2019 [cited by applicant]
US 20060195934A1 · Apuya et al. · 2006 [cited by applicant]
US 20090222957A1 · Apuya et al. · 2009 [cited by applicant]
US 20120090052A1 · Sanz Molinero et al. · 2012 [cited by applicant]
US 20140325710A1 · Abad et al. · 2014 [cited by applicant]
US 20150259699A1 · Nadzan et al. · 2015 [cited by applicant]
US 20160032299A1 · Hashimoto et al. · 2016 [cited by applicant]
US 20170137835A1 · Qu et al. · 2017 [cited by applicant]
US 20180037902A1 · Le et al. · 2018 [cited by applicant]
CN 101824429A · 2010 [cited by applicant]
KR 101460741B1 · 2014 [cited by applicant]
WO 2003020936A1 · 2003 [cited by applicant]
WO 2004013295A2 · 2004 [cited by applicant]
WO 2006109197A2 · 2006 [cited by applicant]
WO WO2010125036A2 · 2010 [cited by examiner]
WO 2015085299A1 · 2015 [cited by applicant]
WO 2015131053A1 · 2015 [cited by applicant]
WO 2018237107A1 · 2018 [cited by applicant]
WO WO2019046756A1 · 2019 [cited by examiner]
Moldoveanu, Serban C., Wayne A. Scott, and Darlene M. Lawson. “Nicotine analysis in several non-tobacco plant materials.” Contributions to Tobacco & Nicotine Research 27.2 (2016): 54-59. (Year: 2016). [cited by examiner]
Keskin, Ozlem, et al. “A new, structurally nonredundant, diverse data set of protein-protein interfaces and its implications.” Protein Science 13.4 (2004): 1043-1055. (Year: 2004). [cited by examiner]
Guo, Haiwei H., Juno Choe, and Lawrence A. Loeb. “Protein tolerance to random amino acid change.” Proceedings of the National Academy of Sciences 101.25 (2004): 9205-9210. (Year: 2004). [cited by examiner]
Thornton, Janet M., et al. “From structure to function: approaches and limitations.” nature structural biology 7.11 (2000): 991-994. (Year: 2000). [cited by examiner]
Köster et al., “RNA-Binding Proteins Revisited—The Emerging [cited by applicant]
Marondedze et al., “The RNA-binding protein repertoire of [cited by applicant]
International Searching Authority in connection with PCT/GB2020/051603 filed Jul. 3, 2020, “The International Search Report and the Written Opinion of the International Searching Authority, or the Declaration”, 19 pages… [cited by applicant]
Ben Saad et al., “AISRG1, a novel gene encoding an RRM-type RNA-binding protein (RBP) from Aeluropus littoralis, confers salt and drought tolerance in transgenic tobacco,” Environmental and Experimental Botany, Mar. 201… [cited by applicant]
Bollenbach et al., “CSP41a, a multifunctional RNA-binding protein, initiates mRNA turnover in tobacco chloroplasts,” The Plant Journal, Nov. 2003, vol. 36, No. 6, pp. 842-852. [cited by applicant]
Hakkinen et al., “Functional characterisation of genes involved in pyridine alkaloid biosynthesis in tobacco,” Phytochemistry, Nov.-Dec. 2007, vol. 68, Issues 22-24, pp. 2773-2785. [cited by applicant]
Rushton et al., “Tobacco Transcription Factors: Novel Insights into Transcriptional Regulation in the Solanaceae,” Plant Physiology, May 2008, vol. 147, Issue 1, pp. 280-295. [cited by applicant]
Voelckel et al., “Herbivore-induced ethylene burst reduces fitness cost of jasmonate- and oral secretion-induced defenses in Nicotiana attenuata,” Oecologia, Jan. 2001, vol. 127, No. 2, pp. 274-280. [cited by applicant]
Edwards et al., “A reference genome for Nicotiana tabacum enables map-based cloning of homeologous loci implicated in nitrogen utilization efficiency,” BMC Genomics, Jun. 19, 2017, vol. 18, No. 448, 14 pages. [cited by applicant]
Feng et al., “Research Progress on Nitrate Accumulation and Regulation During Tobacco Plant Development,” Chinese Journal of Tobacco, Apr. 13, 2019, vol. 25, No. 2, pp. 109-120. [cited by applicant]
Hirose et al., “Nicotiana sylvestris mRNA for RNA-binding glycine-rich protein-1a, complete cds,” Genbank, Jan. 23, 2008, 2 pages. [cited by applicant]
Hirose et al., “RNA-binding glycine-rich protein-1a [Nicotiana sylvestris],” Genbank, Jan. 23, 2008, 1 page. [cited by applicant]
Saurabh et al., “RNA interference: concept to reality in crop improvement,” Planta, Jan. 9, 2014, vol. 239, No. 3, pp. 543-564. [cited by applicant]
Zhao et al., “Progress in the Application of Proteomics Techniques in Tobacco Research,” Chinese Journal of Tobacco, Feb. 28, 2014, vol. 20, No. 1, pp. 103-110. [cited by applicant]