Transcription factors to improve resistance to environmental stress in plants
Plants, plant cells, and seeds are described herein that grow well and are resistant to environmental stresses such as drought and insects, where the plants have one or more mutations that reduce or eliminate the interaction of MYC transcription factors with the JAZ proteins. The plants can have an additional mutation that reduces or eliminates the function of the PHYB gene, and/or a heterologous PIF4 transgene or PIF4 expression cassette to improve the growth of the myc mutant plants. Methods of making and using such plants, plant cells, and seeds are also described.
1. A genetically modified and non-naturally occurring mutant plant cell, a mutant plant, or a mutant plant seed obtained thereof, comprising:
(a) a PhyB loss-of-function mutation comprising at least one mutation to an amino acid in the amino acid sequence as set forth in SEQ ID NO: 33; and
(b) a modified MYC nucleic acid encoding a mutant MYC protein comprising at least one mutation within a JAZ-interacting domain (JID) region of the mutant MYC protein that reduces or eliminates interaction of the mutant MYC protein with JAZ proteins, wherein the JAZ interacting domain (JID) has less than 100% and more than 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 10, and wherein the mutant MYC protein has less than 100% amino acid sequence identity and at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 9, and
wherein the genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof is a transgenic mutant, and wherein expression or lack of expression of the PhyB loss-of-function mutant protein, and expression of the MYC mutant protein rescues wild-type plant biomass and concurrently provides JA (jasmonic acid)-controlled defense in the genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof as compared to a control plant of the same species that is grown under the same conditions, and wherein said JA-controlled defense is resistance to an environmental stress.
2. The genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof, of claim 1 , wherein the PhyB loss-of-function mutation comprises a deletion, substitution, or insertion of a wild-type chromosomal PhyB gene encoding the PhyB polypeptide of SEQ ID NO: 33 so that a truncated PHYB polypeptide, a mutant PHYB polypeptide, or no PHYB polypeptide is expressed.
3. The genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof, of claim 1 , wherein the mutant MYC protein has reduced binding to a JAZ protein from Zea mays as set forth in SEQ ID NO: 60, and wherein the binding is reduced by at least 20% as compared to a corresponding wild type MYC protein that does not have said at least one MYC mutation.
4. The genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof, of claim 1 , wherein the modified MYC nucleic acid is a modification by having a deletion, substitution, or insertion of a chromosomal wild-type MYC gene encoding the protein of SEQ ID NO: 9 to encode the mutant MYC protein.
5. The genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof, of claim 1 , comprising one or more deletions, substitutions, or insertions into one or five wild-type genomic JAZ nucleic acids that encode JAZ proteins with at least 95% amino acid sequence identity to the amino acid sequences as set forth in SEQ ID NO: 60.
6. The genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof, of claim 1 , comprising a loss-of-function chromosomal mutation in an endogenous Zea mays JAZ gene encoding the JAZ protein as set forth in SEQ ID NO: 60.
7. The genetically modified and non-naturally occurring mutant plant cell, the mutant plant, or the mutant plant seed obtained thereof, of claim 1 , wherein the mutant plant or a mutant plant grown from the seed exhibits resistance to the environmental stress compared to a wild type plant of the same species grown under the same environmental conditions, and wherein the environmental stress is insect stress or oxidative stress.
8. A method comprising:
(a) providing one or more genetically modified and non-naturally occurring mutant plant cells comprising a PhyB loss-of-function mutation comprising at least one mutation to an amino acid in the amino acid sequence as set forth in SEQ ID NO: 33;
(b) introducing into at least one of the mutant plant cells at least one transgene or expression cassette encoding a mutant MYC nucleic acid segment that encodes a mutant MYC protein to generate one or more transformed plant cell(s), wherein the mutant MYC protein has less than 100% amino acid sequence identity and at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 9, and wherein the mutant MYC protein comprises at least one mutation within a JAZ-interacting domain (JID) region that reduces or eliminates interaction of the mutant MYC protein with JAZ proteins, wherein the JAZ interacting domain (JID) has less than 100% and more than 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 10; and
(c) generating a genetically modified and non-naturally occurring mutant plant from the one or more transformed plant cells,
wherein expression or lack of expression of the PhyB loss-of-function mutant protein, and expression of the MYC mutant protein rescues wild-type plant biomass and concurrently provides JA (jasmonic acid)-controlled defense in the genetically modified and non-naturally occurring mutant plant as compared to a control plant of the same species that is grown under the same conditions, and wherein said JA-controlled defense is resistance to an environmental stress.
9. The method of claim 8 , further comprising introducing into said at least one of the transformed plant cells one or more deletions, substitutions, or insertions into one or five wild-type genomic JAZ nucleic acids that encode JAZ proteins with at least 95% amino acid sequence identity to the amino acid sequences as set forth in SEQ ID NO: 60.
10. The method of claim 8 , wherein the mutant MYC nucleic acid segment that encodes the mutant MYC protein is a dominant MYC mutation.
11. The method of claim 8 , wherein the mutant MYC protein has reduced binding to a JAZ protein from Zea mays as set forth in SEQ ID NO: 60, and wherein the binding is reduced by at least 20% as compared to a corresponding wild type MYC protein that does not have said at least one MYC mutation.
12. The method of claim 8 , wherein the genetically modified and non-naturally occurring mutant plant exhibits resistance to the environmental stress as compared to a wild type plant of the same species grown under the same environmental conditions, and wherein the environmental stress is insect stress or oxidative stress.