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US 10,501,732 B2Granted
Threonine synthase from nicotiana tabacum and methods and uses thereof
AssigneePhilip Morris Products S.A.
InventorsLucien Bovet, Nicolas Sierro
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Abstract
There is disclosed a mutant, non-naturally occurring or transgenic plant cell comprising: (i) a polynucleotide comprising, consisting or consisting essentially of a sequence encoding a threonine synthase and having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3 or at least 87% sequence identity to SEQ ID NO:4, or SEQ ID NO:5; (ii) a polypeptide encoded by any one of said polynucleotides set forth in (i); or (iii) a polypeptide having at least 95% sequence identity to SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8; or (iv) a construct, vector or expression vector comprising the polynucleotide as set forth in (i).
Claims16
A method for increasing the concentration of free methionine in at least part of a tobacco plant, comprising:
- (a) reducing the expression or activity of threonine synthase in the tobacco plant by introducing into the tobacco plant a polynucleotide that exhibits RNA interference activity against threonine synthase mRNA to generate a mutant, non-naturally occurring, or transgenic tobacco plant, wherein the threonine synthase comprises
- (i) a polynucleotide comprising a sequence encoding the threonine synthase and having at least 97% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5;
- (ii) a polypeptide encoded by any one of said polynucleotides set forth in (i);
- or
- (iii) a polypeptide having at least 99% sequence identity to SEQ ID NO:6, SEQ ID NO:7, OR SEQ ID NO:8;
- (b) measuring the concentration of free methionine in at least a part of the mutant, non-naturally occurring or transgenic tobacco plant obtained in step (a);
- and
- (c) identifying a mutant, non-naturally occurring or transgenic tobacco plant in which the concentration of free methionine therein has increased by at least 50% in comparison to a control tobacco plant in which the polynucleotide or polypeptide in step (a) is not introduced and wherein the visual appearance of said mutant, non-naturally occurring or transgenic tobacco plant is substantially the same as the control tobacco plant three months after field transplant or 36 days after topping.
5 more elements
A method comprising:
- (a) introducing into a tobacco plant a polynucleotide that exhibits RNA interference activity against threonine synthase mRNA to generate a mutant, non-naturally occurring, or transgenic tobacco plant, wherein the threonine synthase mRNA
- (i) is transcribed from a polynucleotide having at least 97% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5;
- or
- (ii) is translated into a polypeptide having at least 99% sequence identity to SEQ ID NO:6, SEQ ID NO:7, OR SEQ ID NO:8;
- (b) measuring the concentration of free methionine in at least a part of the mutant, non-naturally occurring or transgenic tobacco plant obtained in step (a);
- and
- (c) identifying a mutant, non-naturally occurring or transgenic tobacco plant in which the concentration of free methionine therein has increased by at least 50% in comparison to a control tobacco plant in which the polynucleotide or polypeptide in step (a) is not introduced and wherein the visual appearance of said mutant, non-naturally occurring or transgenic tobacco plant is substantially the same as the control tobacco plant three months after field transplant or 36 days after topping.
4 more elements
A method comprising:
- (a) generating a mutant, non-naturally occurring, or transgenic tobacco plant by introducing into a tobacco plant a polynucleotide encoding RNA that
- (i) has at least 97% sequence identity to mRNA transcribed from a polynucleotide comprising a sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5, or
- (ii) has at least 99% sequence identity to mRNA that is translated into a polypeptide comprising an amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, OR SEQ ID NO:8;
- (b) measuring the concentration of free methionine in at least a part of the mutant, non-naturally occurring or transgenic tobacco plant of step (a);
- and
- (c) identifying a mutant, non-naturally occurring or transgenic tobacco plant in which the concentration of free methionine therein has increased by at least 50% in comparison to a control tobacco plant in which the polynucleotide or polypeptide in step (a) is not introduced and wherein the visual appearance of said mutant, non-naturally occurring or transgenic tobacco plant is substantially the same as the control tobacco plant three months after field transplant or 36 days after topping.
3 more elements
Dependencies are inferred from the claim text.