Elevated resistance to insects and plant pathogens without compromising seed production
View Patent ↗Described herein are modified plants, plant cells, and plant seeds that have at least one mutant loss-of-function JAZ gene and at least one loss-of-function cdk8 gene. Such plants are highly resistant to pests and environmental stress and have restored growth and increased seed yield compared to plant lines with a jazD genetic background.
1. A plant, plant cell, or plant seed comprising at least one endogenous cdk8 loss-of-function mutation in one or more endogenous cdk8 genes and endogenous loss-of-function jaz mutations in endogenous jaz genes jaz1, jaz2, jaz5, and jaz6, wherein the endogenous cdk8 genes encode an amino acid sequence having at least 90% identity to SEQ ID NO: 53, 56, 58, 59, 61, 63, or 65 the jaz1 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 1, the jaz2 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 3, 26, 27, or 28, the jaz5 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 9, 37, 38, or 39, and the jaz6 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 11, 40, 41, or 42.
2. The plant, plant cell, or plant seed of claim 1 , further comprising loss-of-function jaz mutations in one or more of the following endogenous jaz genes jaz3, jaz4, jaz7, jaz9, jaz10, or jaz13, wherein
the jaz3 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 5,
the jaz4 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 7,
the jaz7 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 13,
the jaz9 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 15,
the jaz10 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 17, and
the jaz13 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 19.
3. The plant, plant cell, or plant seed of claim 1 , wherein mutation of the endogenous jaz genes reduces endogenous expression of the endogenous jaz gene by at least 50%, or wherein mutation of the one or more endogenous cdk8 genes reduces endogenous expression of the one or more endogenous cdk8 gene by at least 50%.
4. The plant, plant cell, or plant seed of claim 1 , wherein mutation of the endogenous jaz genes reduces endogenous expression of the endogenous jaz genes to undetectable levels, or wherein mutation of the one or more endogenous cdk8 genes reduces endogenous expression of the one or more endogenous cdk8 gene to undetectable levels.
5. The plant, plant cell, or plant seed of claim 1 , wherein the plant, a plant generated from the plant cell, or a plant grown from the plant seed has a seed yield of at least 10% greater than the average seed yield of wild type plants or of unmodified parental plants.
6. The plant, plant cell, or plant seed of claim 1 , wherein the plant, a plant generated from the plant cell, or a plant grown from the plant seed has at least 5% less leaf damage from insect feeding than average damage of insect feeding of a wild type plant of the same species grown for the same time under the same conditions.
7. The plant, plant cell, or plant seed of claim 1 , wherein compared to wild type or an unmodified parental plant line, the plant, a plant generated from the plant cell, or a plant grown from the plant seed has higher levels of defense compounds.
8. The plant, plant cell, or plant seed of claim 1 , wherein compared to wild type or an unmodified parental plant line, the plant, a plant generated from the plant cell, or a plant grown from the plant seed has higher levels of defense compounds that reduce the incidence or number of insect or insect larvae on the plant, plant generated from the plant cell, or plant grown from the seed.
9. The plant, plant cell, or plant seed of claim 1 , wherein compared to wild type or an unmodified parental plant line, the plant, a plant generated from the plant cell, or a plant grown from the plant seed has higher levels of one or more of 3-methylsulphinylpropyl glucosinolate (glucoiberin); 4-methylsulphinylbutyl glucosinolate (glucoraphanin); 5-methyl sulphinylpentyl glucosinolate (glucoalyssin); 6-methylsulphinylhexyl glucosinolate (glucohesperin); 7-methyl sulphinylheptyl glucosinolate (glucoibarin); 3-methylthiopropyl glucosinolate (glucoiberverin); 8-methylsulphinyloctyl glucosinolate (glucohirsutin); 4-methylthiobutyl glucosinolate (glucoerucin); 5-methylthiopentyl glucosinolate (glucoberteroin); or 7-methylthioheptyl glucosinolate.
10. A method comprising cultivating a plant or plant seed comprising at least one endogenous cdk8 loss-of-function mutation in one or more endogenous cdk8 gene(s) and endogenous loss-of-function jaz mutation(s) endogenous jaz genes jaz1, jaz2, jaz5, and jaz6 to produce a mature plant, wherein the endogenous cdk8 gene(s) encode an amino acid sequence having at least 90% identity to SEQ ID NO: 53, 56, 58, 59, 61, 63, or 65, the jaz1 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 1, the jaz2 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 3, 26, 27, or 28, the jaz5 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 9, 37, 38, or 39, and the jaz6 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 11, 40, 41, or 42.
11. The method of claim 10 , further comprising harvesting the mature plant or harvesting seeds, grain, fruit, vegetables, forage, or biomass of the mature plant.
12. The method of claim 10 , wherein the plant or plant seed further comprises loss-of-function jaz mutations in one or more of the following endogenous jaz genes jaz3, jaz4, jaz7, jaz9, jaz10, or jaz13, wherein
the jaz3 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 5,
the jaz4 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 7,
the jaz7 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 13,
the jaz9 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 15,
the jaz10 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 17, and
the jaz13 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 19.
13. The method of claim 10 , wherein mutation of the four or more endogenous jaz genes reduces endogenous expression of the four or more endogenous JAZ gene by at least 50%, or wherein mutation of the one or more endogenous cdk8 genes reduces endogenous expression of the one or more endogenous cdk8 gene by at least 50%.
14. The method of claim 10 , wherein mutation of the four or more endogenous jaz genes reduces endogenous expression of the four or more endogenous jaz to undetectable levels, or wherein mutation of the one or more endogenous cdk8 genes reduces endogenous expression of the one or more endogenous cdk8 gene to undetectable levels.
15. The method of claim 10 , wherein the mature plant has a seed yield of at least 10% greater than the average seed yield of wild type plants or of unmodified parental plants.
16. The method of claim 10 , wherein the mature plant has at least 5% less leaf damage from insect feeding than average leaf damage from insect feeding of wild type plants or of unmodified parental plants of the same species grown for the same time under the same conditions.
17. A method comprising (a) introducing into one or more plant cell(s) at least one chromosomal loss-of-function mutation into at least one endogenous cdk8 gene and into endogenous jaz genes jaz1, jaz2, jaz5, and jaz6, wherein the endogenous cdk8 gene(s) encode an amino acid sequence having at least 90% identity to SEQ ID NO: 53, 56, 58, 59, 61, 63, or 65, the jaz1 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 1, the jaz2 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 3, 26, 27, or 28, the jaz5 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 9, 37, 38, or 39, and the jaz6 gene encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 11, 40, 41, or 42; and (b) generating a plant from the one or more plant cell(s) comprising the at least one chromosomal loss-of-function mutation in the at least one endogenous cdk8 gene and the endogenous jaz genes jaz1, jaz2, jaz5, and jaz6.
18. The method of claim 17 , wherein the plant comprises a deletion of at least a portion of four or more endogenous jaz1, jaz2, jaz3, jaz4, jaz5, jaz6, jaz7, jaz9, jaz10, or jaz13 genes and at least one deletion within a cdk8 gene;
a substitution within at least four or more endogenous jaz1, jaz2, jaz3, jaz4, jaz5, jaz6, jaz7, jaz9, jaz10, or jaz13 genes and at least one substitution within a cdk8 gene, or an insertion into at least four or more endogenous jaz1, jaz2, jaz3, jaz4, jaz5, jaz6, jaz7, jaz9, jaz10, or jaz13 genes and at least one substitution within a cdk8 gene.
19. The method of claim 17 , wherein mutation of the endogenous jaz1, jaz2, jaz5, and jaz6 genes reduces endogenous expression of the endogenous jaz1, jaz2, jaz5, and jaz6 genes by at least 50%, or wherein mutation of the one or more endogenous cdk8 gene(s) reduces endogenous expression of the one or more endogenous cdk8 gene(s) by at least 50%.
20. The method of claim 17 , wherein mutation of the endogenous jaz1, jaz2, jaz5, and jaz6 genes reduces endogenous expression of the endogenous jaz1, jaz2, jaz5, and jaz6 genes to undetectable levels, or wherein mutation of the one or more endogenous cdk8 genes reduces endogenous expression of the one or more endogenous cdk8 gene to undetectable levels.
21. The method of claim 17 , wherein the plant has a seed yield of at least 10% greater than the average seed yield of wild type plants or of unmodified parental plants.
22. The method of claim 17 , wherein the plant has at least 5% less leaf damage from insect feeding than average insect feeding of a wild type plants or of unmodified parental plants of the same species grown for the same time under the same conditions.