Modified brassica plants with increased seed oil content
AssigneeBrookhaven Science Associates, LLC
InventorJorg SCHWENDER
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Abstract
Methods and means are provided to increase the seed oil content of Brassica plants by preventing feedback inhibition by phosphoenolpyruvate (PEP) of a pyrophosphate-dependent phosphofructokinase (PPi-PFK) present in cells of seeds or embryos of these plants, in various manners, including by providing feedback insensitive or less sensitive PPi-PFK.
Claims38
A method to increase oil content in seeds or embryos of a Brassica plant, comprising the step of preventing feedback inhibition by phosphoenolpyruvate (PEP) of a pyrophosphate-dependent phosphofructokinase (PPi-PFK) present in cells of said seeds or said embryos.
A Brassica plant, or seeds thereof, comprising in cells of it seeds or embryos, a pyrophosphate-dependent phosphofructokinase which is less sensitive to said feedback inhibition than a PPi-PFK endogenous to said plant.
A Brassica plant, or seeds thereof comprising a recombinant DNA molecule comprising the following operably linked DNA fragments:
- i. a plant expressible promoter, preferably a seed specific promoter
- ii. a DNA region encoding a PEP carboxylase or PEP carboxykinase ;
- and optionally
- iii. a transcription termination and/or polyadenylation region functional in plant cells.
A chimeric DNA comprising the following operably linked DNA fragments
- a. a plant expressible promoter, preferably a seed-specific promoter;
- b. a DNA region encoding a PPi-PFK which is less sensitive to said feedback inhibition than a PPi-PFK endogenous to said Brassica plant, preferably a DNA region encoding a PPi-PFK comprising an amino acid sequence selected from the amino acid sequences of SEQ ID Nos. 1, 2, 3 or 4, preferably a heterologous DNA region;
- or a DNA region encoding a protein 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with an amino acid sequence selected from the amino acid sequence of SEQ ID Nos. 1,2, 3 or 4, and having PPi-PFK enzymatic activity;
- and optionally
- c. a transcription termination and/or polyadenylation region functional in plant cells.
Use of a PPi-PFK enzyme which is less sensitive to feedback inhibition by PEP than a PPi-PFK endogenous to a Brassica plant to increase the oil content in seeds and or embryos of a Brassica plant.
A method to isolate variants of PPi-PFK enzyme which are less sensitive to feedback inhibition by PEP than a PPi-PFK endogenous to a Brassica plant comprising the steps of
- a. generating a multitude of variant PPi-PFK enzymes from a PEP feedback inhibition sensitive PPi-PFK from a Brassica plant;
- b. identifying the enzymatic activity of each of said variant PPi-PFK enzymes in the presence of PEP;
- c. isolating those enzyme variants which have a greater enzymatic activity in the presence of PEP than the enzymatic activity of said PEP feedback inhibition sensitive PPi-PFK.
A method to isolate a plant cell or plant comprising a variant allele encoding a PPi-PFK variant enzyme which is less sensitive to feedback inhibition by PEP comprising the steps of
- a. providing a population of plant cells or plants, each comprising a multitude of variant PPi-PFK;
- b. identifying the enzymatic activity of each of said PPi-PFK enzymes in the presence of PEP;
- c. isolating those plant cells or plants comprising enzyme variants which have a greater enzymatic activity in the presence of PEP than the enzymatic activity of said feedback inhibition sensitive PPi-PFK.
Dependencies are inferred from the claim text.