Increasing levels of nicotinic alkaloids in plants

Patent metadata

Field	Value
Publication number	US-10190129-B2
Application number	US-201715411721-A
Country	US
Kind code	B2
Filing date	Jan 20, 2017
Priority date	Sep 13, 2006
Publication date	Jan 29, 2019
Grant date	Jan 29, 2019

Abstract

Four genes, A622, NBB1, PMT, and QPT, can be influenced for increasing nicotinic alkaloid levels in Nicotiana plants, as well as for synthesizing nicotinic alkaloids in non-nicotine producing plants and cells. In particular, overexpressing one or more of A622, NBB1, PMT, and QPT may be used to increase nicotine and nicotinic alkaloid levels in tobacco plants. Non-nicotine producing cells can be engineered to produce nicotine and related compounds by overexpressing A622 and NBB1.

First claim

What is claimed is:

1. A method for increasing nicotine and yield in a Nicotiana plant, comprising:
(a) crossing an increased nicotine Nicotiana plant that expresses heterologous NBB1 having the amino acid sequence set forth in SEQ ID NO: 2 and A622 having the amino acid sequence set forth in SEQ ID NO: 4 with a high yielding Nicotiana plant; and
(b) selecting progeny plant with increased nicotine and high yield, wherein the plant has an increased nicotine content and yield relative to a non-transformed control plant.

2. The method according to claim 1 , wherein the increased nicotine Nicotiana plant is produced by:
(a) transforming a Nicotiana plant with a construct comprising, in the 5′ to 3′ direction, a promoter operably linked to a heterologous nucleic acid molecule selected from:
(i) the nucleotide sequence set forth in SEQ ID NO: 1, or
(ii) a nucleotide sequence that encodes the polypeptide having the amino acid sequence set forth in SEQ ID NO: 2; and
(iii) the nucleotide sequence set forth in SEQ ID NO: 3, or
(iv) a nucleotide sequence that encodes the polypeptide having the amino acid sequence set forth in SEQ ID NO: 4;

(b) regenerating transgenic Nicotiana plants from the transformed Nicotiana plant; and
(c) selecting a transgenic Nicotiana plant having increased-nicotine content relative to a control plant,
wherein the plant has an increased nicotine content relative to a non-transformed control plant.

3. An increased nicotine and yield plant produced by the method of claim 2 .

4. An increased nicotinic alkaloid product produced from the plant of claim 3 , wherein:
(a) the product is selected from the group consisting of a cigarette, cigarette tobacco, a cigar, cigar tobacco, pipe tobacco, chewing tobacco, snuff, snus, leaf tobacco, shredded tobacco, cut tobacco, a pharmaceutical and a nutraceutical, and
(b) the product comprises the construct.

5. A method for increasing nicotine and yield in a Nicotiana plant, comprising:
(a) transforming a Nicotiana plant with:
(i) a first construct comprising, in the 5′ to 3′ direction, a promoter operably linked to a heterologous nucleic acid molecule selected from: the nucleotide sequence set forth in SEQ ID NO: 1, or a nucleotide sequence that encodes the polypeptide having the amino acid sequence set forth in SEQ ID NO: 2; and the nucleotide sequence set forth in SEQ ID NO: 3, or a nucleotide sequence that encodes the polypeptide having the amino acid sequence set forth in SEQ ID NO: 4; and
(ii) a second construct comprising, in the 5′ to 3′ direction, a promoter operably linked to a heterologous nucleic acid encoding an enzyme that increases yield;

(b) regenerating transgenic Nicotiana plants from the transformed Nicotiana plant; and
(c) selecting a transgenic Nicotiana plant having increased-nicotine content and increased yield relative to a control plant,
wherein the plant has an increased nicotine content and yield relative to a non-transformed control plant.

6. An increased nicotine and yield plant produced by the method of claim 5 .

7. An increased nicotinic alkaloid product produced from the plant of claim 6 , wherein the product is selected from the group consisting of a cigarette, cigarette tobacco, a cigar, cigar tobacco, pipe tobacco, chewing tobacco, snuff, snus, leaf tobacco, shredded tobacco, cut tobacco, a pharmaceutical and a nutraceutical, wherein the product comprises the first and second constructs.

Assignees

• 22Nd Century Ltd Llc

Inventors

• Hashimoto Takashi
• Kajikawa Masataka

Classifications

• A24B13/00

  Tobacco for pipes, for cigars, e.g. cigar inserts, or for cigarettes; Chewing tobacco; Snuff (reconstituted tobacco products A24B3/14) · CPC title

• C12N15/8243 Primary

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

• A24B13/02

  Flakes or shreds of tobacco · CPC title

• A61K31/465

  Nicotine; Derivatives thereof · CPC title

• A24D1/00

  Cigars; Cigarettes · CPC title