Digestibility of plant biomass
Plants described herein have increased biomass and are more readily digested into fermentable sugars when the plants express increased levels of one or more types of CGR2 and/or CGR3 enzymes.
1. A plant comprising: a heterologous expression cassette comprising a heterologous promoter operably linked to an isolated nucleic acid segment encoding a methyltransferase CGR2 or CGR3 enzyme with at least 95% amino acid sequence identity to any of SEQ ID NO: 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 or 24, wherein the plant's pectin has at least 10% more methylesters than a wild type plant of the same species that does not have the expression cassette.
2. The plant of claim 1 , wherein the expression cassette expresses the CGR2 enzyme and/or the CGR3 enzyme, or can be induced to express the CGR2 enzyme and/or the CGR3 enzyme, at levels sufficient to increase the plant's biomass by at least 5% compared to a wild type plant of the same species that does not, comprise the expression cassette.
3. The plant of claim 1 , wherein enzymatic digestion of biomass from the plant releases at least 10% more fermentable sugar than is released than from a wild type plant biomass of the same species that does not have the expression cassette.
4. The plant of claim 1 , wherein the heterologous promoter is a plant gene promoter, a bacterial gene promoter, a plant housekeeping gene promoter, a tissue-specific promoter, or an inducible promoter.
5. The plant of claim 1 , wherein the heterologous promoter is a dermal tissue-specific promoter, a vascular tissue-specific promoter, or a ground tissue-specific promoter.
6. The plant of claim 1 , wherein the heterologous promoter is a cauliflower mosaic virus promoter, a CaMV 35S promoter, a CaMV 19S promoter, a nos promoter, an Adh 1 promoter, a sucrose synthase promoter, an a-tubulin promoter, a ubiquitin promoter, an actin promoter, an actin promoter from rice, a cab promoter, a PEPCase promoter, an R gene complex promoter, a poplar xylem-specific secondary cell wall specific cellulose synthase 8 promoter, a Z10 promoter from a gene encoding a 10 kD zein protein, a Z27 promoter from a gene encoding a 27 kD zein protein, a light inducible promoter from the pea rbcS gene, or a phaseolin promoter from beans.
7. The plant of claim 1 , wherein the plant is an oil-producing plant, a starch plant, a forage plant, a vegetable plant, a grain-producing plant, straw-producing plant, a grass plant, a woody plant, a softwood, a hardwood, a gymnosperm, or a legume.
8. A plant biomass from the plant of claim 1 .
9. A seed from the plant of claim 1 , wherein the seed comprises the heterologous expression cassette of claim 1 .
10. A method comprising digesting biomass from the plant of claim 1 to yield fermentable sugars.
11. A method comprising:
obtaining plant biomass from a plant comprising an expression cassette comprising an isolated nucleic acid segment encoding a methyltransferase CGR2 or CGR3 enzyme with at least 95% amino acid sequence identity to any of SEQ ID NO: 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 or 24; and
digesting the plant biomass to yield at least 10% more fermentable sugars than is recovered than from a wild type plant biomass of the same species that does not have the expression cassette and is digested by the same procedure.
12. The method of either claim 10 , wherein pectin in the plant biomass has at least 5% more methylesters than a wild type plant of the same species that does not have the expression cassette.
13. A method comprising:
expressing a methyltransferase CGR2 and/or CGR3 enzyme with at least 95% amino acid sequence identity to any of SEQ ID NO: 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23 or 24 from a heterologous expression cassette at levels sufficient to increase a plant's biomass by at least 5% compared to a wild type plant of the same species that does not comprise the expression cassette;
wherein the expression cassette comprises at least one heterologous promoter and an isolated nucleic acid segment encoding a CGR2 enzyme and/or an isolated nucleic acid segment encoding a CGR3 enzyme.
14. The method of claim 10 , wherein the heterologous promoter for the CGR2 expression cassette or the CGR3 expression cassette is independently selected from a plant gene promoter, a bacterial gene promoter, a plant housekeeping gene promoter, a tissue-specific promoter, or an inducible promoter.
15. The method of claim 10 , wherein the heterologous promoter for the CGR2 expression cassette or the CGR3 expression cassette is independently selected from a dermal tissue-specific promoter, a vascular tissue-specific promoter, or a ground tissue-specific promoter, a cell-wall tissue-specific promoter, a leaf tissue-specific promoter, a xylem tissue-specific promoter, a phloem-specific promoter, a collenchyma cell-specific promoter, a parenchyma-specific promoter, a seed specific promoter, or a meristematic-specific promoter.
16. The method of claim 10 , wherein the plant is an oil-producing plant, a starch plant, a forage plant, a vegetable plant, a grain-producing plant, straw-producing plant, a grass plant, a woody plant, a softwood, a hardwood, a gymnosperm, or a legume.
17. The method of claim 10 , wherein the plant is canola, potato, lupin, sunflower, cottonseed, alfalfa, clover, fescue, cucumber, tomato, maize, wheat, barley, oats, rice, sorghum, millet, rye, switchgrass, prairie grass, wheat grass, sudangrass, sorghum, soybean, a poplar, a pine, a eucalyptus , a loblolly pine, a Jack pine, a Southern pine, a Radiata pine, a spruce, a Douglas fir, aspen, miscanthus , willow, bromegrass, or Western.