Compositions and methods for producing tobacco plants and products having altered alkaloid levels

Patent metadata

Field	Value
Publication number	US-2022010324-A1
Application number	US-202117337161-A
Country	US
Kind code	A1
Filing date	Jun 2, 2021
Priority date	Jun 3, 2020
Publication date	Jan 13, 2022
Grant date	—

Abstract

The present disclosure provides tobacco having genetic modification(s) in a group of ERF genes. Also provided are tobacco plants with altered total alkaloid and nicotine levels and commercially acceptable leaf grade, their development via breeding or transgenic approaches, and production of tobacco products from these tobacco plants. Further provided are compositions and methods for producing tobacco plants having novel mutations or alleles to reduce nicotine levels. Further provided are sequence polymorphisms and molecular markers for breeding tobacco with reduced nicotine or alkaloids while maintaining tobacco leaf grade and tobacco product quality.

First claim

1 . A modified tobacco plant, or part thereof, comprising a non-natural mutation in a polynucleotide having at least 90% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 3, 13, 14, 17, 18, 19, 38, 48, 49, 52-54, 153, 154, 158, 159, 202, 203, 204, and 205.

2 . The modified tobacco plant, or part thereof, of claim 1 , wherein said polynucleotide has at least 95% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 3, 13, 14, 17, 18, 19, 38, 48, 49, 52, 53, 54, 153, 154, 158, 159, 202, 203, 204, and 205.

3 . The modified tobacco plant, or part thereof, of claim 1 , wherein said polynucleotide has 100% identity to a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 3, 13, 14, 17, 19, 19, 38, 48, 49, 52, 53, 54, 153, 154, 158, 159, 202, 203, 204, and 205.

4 . The modified tobacco plant, or part thereof, of claim 1 , wherein said at least one modified tobacco plant comprises a reduced amount of at least one alkaloid as compared to a control tobacco plant lacking said mutation.

5 . The modified tobacco plant, or part thereof, of claim 4 , wherein said at least one alkaloid is selected from the group consisting of anabasine, anatabine, nicotine, and nornicotine.

6 . A modified tobacco plant, or part thereof, comprising a non-natural mutation in a polynucleotide having a nucleic acid sequence encoding a polypeptide having at least 90% identity or similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 73, 83, 84, 87, 88, 89, 180, 181, 206, and 207.

7 . The modified tobacco plant, or part thereof, of claim 6 , wherein said polynucleotide has a nucleic acid sequence encoding a polypeptide having at least 95% identity or similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 73, 83, 84, 87, 88, 89, 180, 181, 206, and 207.

8 . The modified tobacco plant, or part thereof, of claim 6 , wherein said polynucleotide has a nucleic acid sequence encoding a polypeptide having 100% identity or similarity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 73, 83, 84, 87, 88, 89, 180, 181, 206, and 207.

9 . The modified tobacco plant, or part thereof, of claim 6 , wherein said modified tobacco plant comprises a reduced level of at least one alkaloid as compared to a control tobacco plant lacking said mutation relative to a control plant not having said genetic modification or mutation or recombinant nucleic acid construct.

10 . The modified tobacco plant, or part thereof, of claim 6 , wherein said higher USDA grade index value is at least 5% higher than that of said comparable leaf of said control plant.

11 . The tobacco plant, or part thereof, of claim 9 , wherein said at least one alkaloid is selected from the group consisting of anabasine, anatabine, nicotine, and nornicotine.

12 . Cured tobacco material from the tobacco plant of claim 6 .

13 . A tobacco product comprising the cured tobacco material of claim 12 .

14 . The tobacco product of claim 13 , wherein said tobacco product is a selected from the group consisting of a cigarette, a cigarillo, a non-ventilated recess filter cigarette, a vented recess filter cigarette, a cigar, snuff, pipe tobacco, cigar tobacco, cigarette tobacco, chewing tobacco, leaf tobacco, shredded tobacco, and cut tobacco, reconstituted tobacco, loose leaf chewing tobacco, plug chewing tobacco, moist snuff, snus, and nasal snuff, or a smokeless tobacco product.

15 . A method of producing a modified tobacco plant comprising:
(a) inducing a non-natural mutation in at least one tobacco cell in an endogenous nucleic acid sequence encoding a polypeptide comprising an amino acid sequence at least 90% identical or similar to an amino acid sequence selected from the group consisting of SEQ ID NOs: 73, 83, 84, 87, 88, 89, 180, 181, 206, and 207; (b) selecting at least one tobacco cell comprising said non-natural mutation from step (a); and (c) regenerating at least one modified tobacco plant from said at least one tobacco cell selected in step (b);
wherein said at least one modified tobacco plant comprises a reduced amount of at least one alkaloid as compared to a control tobacco plant lacking said mutation.

16 . The method of claim 15 , wherein said at least one modified tobacco plant comprises a reduced amount of at least one alkaloid as compared to a control tobacco plant lacking said mutation.

17 . The method of claim 15 , wherein said endogenous nucleic acid sequence is at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 3, 13, 14, 17, 18, 19, 38, 48, 49, 52, 53, 54, 153, 154, 158, 159, 202, 203, 204, and 205.

18 . The method of claim 15 , wherein said at least one alkaloid is selected from the group consisting of anabasine, anatabine, nicotine, and nornicotine.

19 . The method of claim 15 , wherein said method further comprises:
(d) growing said modified tobacco plant regenerated in step (c).

20 . The method of claim 19 , wherein said method further comprises:
(e) crossing said modified tobacco plant grown in step (d) with a second tobacco plant; and (f) obtaining at least one seed from said crossing in step (e).

Assignees

• Altria Client Services Llc

Inventors

• Pramod Sreepriya
• De Godoy Lusso Marcos Fernando
• Frederick Jesse
• Adams Andrew
• Xu Dongmei

Classifications

• C07K14/415

  from plants · CPC title

• A24B15/10

  Chemical features of tobacco products or tobacco substitutes · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N15/8243 Primary

  involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine · CPC title

• A01H6/823

  Nicotiana, e.g. tobacco · CPC title