Enhanced stability engineered WRINKLED1 transcription factor
The application describes stabilized WRINKLED1 transcription factors, as well as nucleic acids, and expression cassettes that encode and express such stabilized WRINKLED1 transcription factors and that are useful for increasing production of oils in plants and seeds.
1. A modified transcription factor having increased stability, comprising:
a protein comprising amino acid sequence SEQ ID NO: 1 modified to have a deletion of at least 10 amino acids between amino acid positions 368-430 of SEQ ID NO:1, or a protein comprising sequence SEQ ID NO:1 modified to have one or more substitutions of the X residues of SEQ NO:4;
a protein comprising amino acid sequence SEQ ID NO:5 modified to have a deletion of at least 10 amino acids within C-terminal sequence SEQ ID NO:7, or a protein comprising SEQ NO:5 modified to have one or more substitutions of the X residues of SEQ ID NO:8;
a protein comprising amino acid sequence SEQ ID NO:9 modified to have a deletion of at least 10 amino acids within C-terminal sequence SEQ ID NO:11, or a protein comprising SEQ ID NO:9 modified to have one or more substitutions of the X residues of SEQ ID NO:12;
a protein comprising amino acid sequence SEQ ID NO:13 modified to have a deletion of at least 10 amino acids within C-terminal sequence SEQ ID NO:15, or a protein comprising SEQ ID NO:13 modified to have one or more substitutions of the X residues of SEQ ID NO:16;
a protein comprising amino acid sequence SEQ ID NO:17 modified to have a deletion of at least 10 amino acids within C-terminal sequence SEQ ID NO:19, or a protein comprising SEQ ID NO:17 modified to have one or more substitutions of the X residues of SEQ ID NO:20;
a protein comprising amino acid sequence SEQ ID NO:21 modified to have a deletion of at least 10 amino acids within C-terminal sequence SEQ NO:23, or a protein comprising SEQ NO:21 modified to have one or more substitutions of the X residues of SEQ ID NO:24; or
a protein comprising amino acid sequence SEQ ID NO:25 modified to have a deletion of at least 10 amino acids within C-terminal sequence SEQ ID NO:27, or a protein comprising SEQ ID NO:25 modified to have one or more substitutions of the X residues of SEQ ID NO:28;
wherein at least 1.5-fold increased oil content is present in plant tissues when the modified transcription factor is expressed in the plant tissues, and wherein each X residue is separately alanine, glycine, valine, leucine, isoleucine, or methionine.
2. The modified transcription factor of claim 1 comprising:
a. an AtWRI1 1-397 protein with a sequence consisting of SEQ ID NO:67;
b. an IDR3-PEST domain having SEQ ID NO:61;
c. an IDR3-PEST domain having SEQ ID NO:63; or
d. a protein comprising SEQ ID NO:62.
3. The modified transcription factor of claim 1 , wherein one or more deletion is of at least 30 amino acids.
4. A nucleic acid encoding the modified transcription factor of claim 1 .
5. An expression cassette comprising a promoter operably linked to a nucleic acid encoding the modified transcription factor of claim 1 .
6. A plant comprising an expression cassette comprising a promoter operably linked to a nucleic acid encoding the modified transcription factor of claim 1 .
7. The plant of claim 6 , wherein the plant is a food plant, vegetable oil plant, seed oil plant, forage plant, or fodder plant.
8. A plant seed comprising an expression cassette comprising a promoter operably linked to a nucleic acid encoding the modified transcription factor of claim 1 .
9. The plant seed of claim 8 , wherein the plant seed is from a food plant, vegetable oil plant, seed oil plant, forage plant, or fodder plant.
10. A method of generating oil comprising isolating tissues or seeds from the plant of claim 6 , and extracting oil from the isolated tissues or seeds.
11. A method of generating oil comprising extracting oil from the seed(s) of claim 8 .