Inhibition of bolting and flowering of a beta vulgaris plant

Patent metadata

Field	Value
Publication number	US-11034971-B2
Application number	US-201615771873-A
Country	US
Kind code	B2
Filing date	Oct 28, 2016
Priority date	Oct 30, 2015
Publication date	Jun 15, 2021
Grant date	Jun 15, 2021

Abstract

The present invention provides means for inhibiting the bolting and flowering of a Beta vulgaris plant, including an isolated nucleic acid, which can be used to produce a transgenic Beta vulgaris plant, where bolting and flowering is inhibited after vernalization. Furthermore, the invention discloses vectors, transgenic and non-transgenic, non-bolting plants and parts thereof, and methods for producing such plants.

First claim

The invention claimed is:

1. A method for producing a Beta vulgaris plant, where the bolting and flowering is inhibited after vernalization, comprising
either the following steps (I), (II) and (III):
(I) mutagenizing one or more parts of a Beta vulgaris plant and subsequently regenerating one or more Beta vulgaris plants from the one or more parts to yield a regenerated plant, or mutagenizing one or more Beta vulgaris plants to yield a mutagenized plant;
(II) identifying a regenerated or mutagenized plant of (I) that exhibits one or more mutations in a first endogenous DNA sequence and exhibits one or more mutations in a second endogenous DNA sequence; and
(III) generating a Beta vulgaris plant in which the one or more mutations in the first endogenous DNA sequence and the one or more mutations in the second endogenous DNA sequence are homozygous,

or the following steps (IV), (V), (VI) and (VII):
(IV) mutagenizing one or more parts of a Beta vulgaris plant and subsequently regenerating one or more Beta vulgaris plants from the one or more parts to yield a regenerated plant, or mutagenizing one or more Beta vulgaris plants to yield a mutagenized plant;
(V) identifying a first regenerated or mutagenized plant of (III) that exhibits one or more mutations in a first endogenous DNA sequence and a second regenerated or mutagenized plant of (III) that exhibits one or more mutations in a second endogenous DNA sequence;
(VI) crossing the first regenerated or mutagenized plant with the second regenerated or mutagenized plant and selecting a progeny comprising the one or more mutations in the first endogenous DNA sequence and/or in a regulatory sequence thereof and the one or more mutations in the second endogenous DNA sequence; and
(VII) generating from the progeny of (VI) a Beta vulgaris plant in which the one or more mutations in the first endogenous DNA sequence and the one or more mutations in the second endogenous DNA sequence are homozygous;

wherein the first endogenous DNA sequence has a nucleic acid sequence identical to a sequence that
a) exhibits a sequence comprising nucleotides 1-185, 6923-7001, 7589-7653, 8561-8659, 11792-11834, 21139-21181, 21286-21453, and 21569-21668 of SEQ ID NO: 1 or comprising the coding sequence of SEQ ID NO: 2,
b) comprises a nucleotide sequence exhibiting at least 98% identity to nucleotides 1-185, 6923-7001, 7589-7653, 8561-8659, 11792-11834, 21139-21181, 21286-21453, and 21569-21668 of SEQ ID NO: 1 or comprises a nucleotide sequence exhibiting at least 98% identity to the coding sequence of SEQ ID NO: 2,
c) is complementary to any one of the sequences recited in (a) and (b) above
d) encodes a protein comprising an amino acid sequence exhibiting at least 98% sequence identity to SEQ ID NO: 5;

wherein the second endogenous DNA sequence has a nucleic acid sequence identical to a sequence that
A) exhibits a sequence comprising SEQ ID NO: 3 or comprising the coding sequence of SEQ ID NO: 4,
B) comprises a nucleotide sequence exhibiting at least 98% identity to nucleotides 1-185, 19324-19462, 19501-19565, 19643-19742, 28158-28199, 28279-28320, 28420-28532, and 29128-29237 of SEQ ID NO: 3 or comprises a nucleotide sequence exhibiting at least 98% identity to the coding sequence of SEQ ID NO: 4,
C) is complementary to any one of the sequences recited in (A) and (B) above, or
D) encodes a protein comprising an amino acid sequence exhibiting at least 98% sequence identity to SEQ ID NO: 6.

2. A Beta vulgaris plant, produced by the method of claim 1 , or a part thereof,
wherein the part is selected from the group consisting of a stem, a root, and a seed, and
wherein the bolting and flowering of the plant is inhibited after vernalization.

3. A Beta vulgaris plant, where the bolting and flowering is inhibited after vernalization, wherein the plant comprises one or more mutations in a first endogenous DNA sequence and comprises one or more mutations in a second endogenous DNA sequence,
wherein the first endogenous DNA sequence comprises a nucleic acid sequence identical to a sequence that
a) comprises a sequence comprising nucleotides 1-185, 6923-7001, 7589-7653, 8561-8659, 11792-11834, 21139-21181, 21286-21453, and 21569-21668 of SEQ ID NO: 1 or comprising the coding sequence of SEQ ID NO: 2,
b) comprises a nucleotide sequence exhibiting at least 98% identity to nucleotides 1-185, 6923-7001, 7589-7653, 8561-8659, 11792-11834, 21139-21181, 21286-21453, and 21569-21668 of SEQ ID NO: 1 or comprises a nucleotide sequence exhibiting at least 98% identity to the coding sequence of SEQ ID NO: 2,
c) is complementary to any one of the sequences recited in (a) and (b) above
d) encodes a protein comprising an amino acid sequence exhibiting at least 98% sequence identity to SEQ ID NO: 5;

wherein the second endogenous DNA sequence comprises a nucleic acid sequence identical to a sequence that
A) comprises a sequence comprising SEQ ID NO: 3 or comprising the coding sequence of SEQ ID NO: 4,
B) comprises a nucleotide sequence exhibiting at least 98% identity to nucleotides 1-185, 19324-19462, 19501-19565, 19643-19742, 28158-28199, 28279-28320, 28420-28532, and 29128-29237 of SEQ ID NO: 3 or comprises a nucleotide sequence exhibiting at least 98% identity to the coding sequence of SEQ ID NO: 4,
C) is complementary to any one of the sequences recited in (A) and (B) above, or
D) encodes a protein comprising an amino acid sequence exhibiting at least 98% sequence identity to SEQ ID NO: 6;

wherein the one or more mutations cause a reduction of the activity or stability of the protein or polypeptide encoded by the endogenous DNA sequence compared to a non-mutagenized wildtype plant.

Assignees

• Kws Saat Se & Co Kgaa

Inventors

• De Maagd Rudolf Aart
• Van Arkel Jeroen
• Angenent Gerrit Cornelis
• Wurbs David
• Kraus Josef

Classifications

• C07K14/415 Primary

  from plants · CPC title

• A01H6/024

  Beta vulgaris [beet] · CPC title

• C12N15/8218

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

• C12N15/8222

  Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation · CPC title

• C12N15/8265 Primary

  Transgene containment, e.g. gene dispersal · CPC title