Tissue specific reduction of lignin
View Patent ↗The present invention provides an expression cassette comprising a polynucleotide that encodes a protein that diverts a monolignol precursor from a lignin biosynthesis pathway in the plant, which is operably linked to a heterologous promoter. Also provided are methods of engineering a plant having reduced lignin content, as well as plant cells, plant parts, and plant tissues from such engineered plants.
1. A method of engineering a plant having reduced lignin content, the method comprising:
introducing into the plant an expression cassette comprising a polynucleotide that encodes a bacterial shikimate kinase, wherein the polynucleotide is operably linked to a heterologous promoter, and further wherein the promoter is a secondary cell wall-specific promoter, a fiber cell-specific promoter, or a promoter from a gene in the lignin biosynthesis pathway; and
culturing the plant under conditions in which the shikimate kinase is expressed.
2. The method of claim 1 , wherein the polynucleotide encodes a Mycobacterium tuberculosis shikimate kinase (AroK).
3. The method of claim 1 , wherein the promoter is a secondary cell wall-specific promoter or a fiber cell-specific promoter.
4. The method of claim 3 , wherein the promoter is an IRX5 promoter.
5. The method of claim 1 , wherein the promoter is from a gene in the lignin biosynthesis pathway.
6. The method of claim 5 , wherein the promoter is a cinnamate 4-hydroxylase (C4H), coumarate 3-hydroxylase (C3H), hydroxycinnamoyl-Coenzyme A shikimate/quinate hydroxycinnamoyltransferase (HCT), cinnamoyl-CoA reductase (CCR1), cinnamyl alcohol dehydrogenase 4 (CAD4), cinnamyl alcohol dehydrogenase 5 (CAD5), ferulate 5-hydroxylase (F5H), phenylalanine ammonia-lyase 1 (PAL1), phenylalanine ammonia-lyase 2 (PAL2), 4-coumarate CoA ligase 1 (4CL1), or caffeoyl/CoA-3-O-methyltransferase (CCoAMT) promoter.
7. The method of claim 6 , wherein the promoter is a C4H promoter.
8. The method of claim 1 , wherein the plant is selected from the group consisting of Arabidopsis , poplar, eucalyptus, rice, corn, switchgrass, sorghum, millet, miscanthus, sugarcane, pine, alfalfa, wheat, soy, barley, turfgrass, tobacco, hemp, bamboo, rape, sunflower, willow, and Brachypodium.
9. A plant engineered by the method of claim 1 .
10. A plant cell from the plant of claim 9 , wherein plant cell comprises the expression cassette.
11. A seed, flower, leaf, or fruit from the plant of claim 9 , wherein the seed, flower, leaf, or fruit comprises the expression cassette.
12. A plant cell comprising a polynucleotide that encodes a bacterial shikimate kinase, wherein the polynucleotide is operably linked to a heterologous promoter, and wherein the promoter is a secondary cell wall-specific promoter, a fiber cell-specific promoter, or a promoter from a gene in the lignin biosynthesis pathway.
13. The plant cell of claim 12 , wherein the bacterial shikimate kinase is Mycobacterium tuberculosis shikimate kinase (AroK).
14. The plant cell of claim 12 , wherein the promoter is a cinnamate 4-hydroxylase (C4H), coumarate 3-hydroxylase (C3H), hydroxycinnamoyl-Coenzyme A shikimate/quinate hydroxycinnamoyltransferase (HCT), cinnamoyl-CoA reductase (CCR1), cinnamyl alcohol dehydrogenase 4 (CAD4), cinnamyl alcohol dehydrogenase 5 (CAD5), ferulate 5-hydroxylase (F5H), phenylalanine ammonia-lyase 1 (PAL1), phenylalanine ammonia-lyase 2 (PAL2), 4-coumarate CoA ligase 1 (4CL1), or caffeoyl/CoA-3-O-methyltransferase (CCoAMT) promoter.
15. The plant cell of claim 14 , the promoter is a C4H promoter.
16. A plant comprising the plant cell of claim 13 , wherein the plant has reduced lignin content localized to the secondary cell wall tissue or fiber cells of the plant.