Temporally and spatially targeted dynamic nitrogen delivery by remodeled microbes
Inventors
Tamsir, Alvin • BLOCH, Sarah • REISINGER, MARK • SANDERS, ERNEST • BROGLIE, RICHARD • CLARK, Rosemary • Temme, Karsten
Assignees
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Publication Number
US-12391624-B2
Publication Date
2025-08-19
Expiration Date
Abstract
The present disclosure provides non-intergeneric remodeled microbes that are able to fix atmospheric nitrogen and deliver such to plants in a targeted, efficient, and environmentally sustainable manner. The utilization of the taught microbial products will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived nitrogen fertilizer, by mitigating or eliminating the need for exogenous nitrogen-containing fertilizers. The remodeled microbes have unique colonization and nitrogen fixation abilities, which enable the microbes to deliver nitrogen to a cereal plant in a spatially targeted (e.g. rhizospheric) and temporally targeted (e.g. during advantageous stages of plants life cycle) manner. The microbes are able to replace the standard agricultural practice of sidedressing and enable a more environmentally sustainable form of farming. The present disclosure also provides methods of using non-intergeneric remodeled microbes, for example, to fix atmospheric nitrogen by reducing or eliminating the need for exogenous nitrogen-containing fertilizers, to increase yield, and to reduce infield variability in the yield.
Core Innovation
The invention provides fixed atmospheric nitrogen to a cereal plant by providing to a locus a plurality of non-intergeneric remodeled bacteria. The bacteria each produce fixed N, colonize the root surface of the plurality of cereal plants, and supply the cereal plants with fixed N. The bacteria produce fixed N in the aggregate at least about 15 pounds of fixed N per acre over the course of at least about 10 days to about 60 days.
The remodeled bacteria include at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, of the nitrogen fixation or assimilation genetic regulatory network. The genetic variation results in increased expression or activity of NifA or glutaminase, decreased expression or activity of NifL, NtrB, glutamine synthetase, GlnB, GlnK, DraT, or AmtB, decreased adenylyl-removing activity of GlnE, or decreased uridylyl-removing activity of GlnD.
In corn-focused embodiments, the invention increases corn yield per acre in agriculturally challenging soil and reduces infield variability for corn yield per acre. The methods include providing non-intergeneric remodeled bacteria that colonize the root surface of a plurality of corn plants and supply fixed N, where agriculturally challenging soil includes at least about 30% sand. Yield increase is defined relative to a control plurality of corn plants, and infield variability is evaluated by a lower standard deviation of corn mean yield measured across the locus compared to control.
Claims Coverage
The consolidated disclosure identifies three independent claims, each centered on non-intergeneric remodeled bacteria that colonize cereal, including corn, root surfaces and provide fixed atmospheric nitrogen, with claim-specific agronomic endpoints. Across the independent claims, the inventive features cover fixed-N provision by engineered bacteria with specified regulatory-network modifications, corn yield increase in agriculturally challenging soil with a defined yield threshold, and reduced infield yield variability measured by standard deviation compared to a control.
Non-intergeneric remodeled bacteria engineered for fixed atmospheric nitrogen delivery to cereals
A method of providing fixed atmospheric nitrogen to a cereal plant by providing to a locus a plurality of non-intergeneric remodeled bacteria that each produce fixed N and comprise at least one genetic variation introduced into at least one gene, or non-coding polynucleotide, of the nitrogen fixation or assimilation genetic regulatory network, resulting in increased expression or activity of NifA or glutaminase, decreased expression or activity of NifL, NtrB, glutamine synthetase, GlnB, GlnK, DraT, AmtB, decreased adenylyl-removing activity of GlnE, or decreased uridylyl-removing activity of GlnD.
Root-surface colonization and aggregate fixed n output
Said plurality of non-intergeneric remodeled bacteria colonize the root surface of said plurality of cereal plants and supply the cereal plants with fixed N, and wherein said plurality of non-intergeneric remodeled bacteria produce in the aggregate at least about 15 pounds of fixed N per acre over the course of at least about 10 days to about 60 days.
Corn yield increase in agriculturally challenging soil
A method for increasing corn yield per acre in agriculturally challenging soil, comprising providing to a locus located in an agriculturally challenging soil a plurality of non-intergeneric remodeled bacteria that each produce fixed N and providing to the locus located in an agriculturally challenging soil a plurality of corn plants, wherein said plurality of non-intergeneric remodeled bacteria colonize the root surface of said plurality of corn plants and supply the corn plants with fixed N, wherein said agriculturally challenging soil comprises a soil that comprises at least about 30% sand, and wherein said plurality of corn plants achieve at least a 1 bushel per acre yield increase, as compared to a control plurality of corn plants when the control plurality of corn plants is provided to the locus.
Reducing infield variability of corn yield per acre
A method for reducing infield variability for corn yield per acre, comprising providing to a locus a plurality of non-intergeneric remodeled bacteria that each produce fixed N and providing to the locus a plurality of corn plants, wherein said plurality of non-intergeneric remodeled bacteria colonize the root surface of said plurality of corn plants and supply the corn plants with fixed N, and wherein the standard deviation of corn mean yield measured across the locus as measured in bushels per acre is lower for the plurality of corn plants colonized by said non-intergeneric remodeled bacteria, as compared to a control plurality of corn plants when the control plurality of corn plants is provided to the locus.
Across the independent claims, the inventive coverage centers on applying a plurality of non-intergeneric remodeled bacteria that colonize cereal or corn root surfaces and supply fixed atmospheric nitrogen. The bacteria are engineered with genetic variation in the nitrogen fixation or assimilation genetic regulatory network to produce defined functional shifts, including increased NifA or glutaminase function and/or decreased NifL, NtrB, and related activities, including adenylyl- and uridylyl-removing activities. For corn, the independent claims further specify yield increase in agriculturally challenging soil and reduced infield variability versus control.
Stated Advantages
Provides fixed atmospheric nitrogen to cereal plants by colonizing root surfaces and supplying fixed N.
Increases corn yield per acre in agriculturally challenging soil by at least about 1 bushel per acre compared to control.
Reduces infield variability for corn yield per acre by lowering the standard deviation of corn mean yield across the locus compared to control.
Documented Applications
Providing fixed atmospheric nitrogen to a cereal plant by applying non-intergeneric remodeled bacteria that colonize the root surface and supply fixed N.
Increasing corn yield per acre in agriculturally challenging soil comprising at least about 30% sand, compared to a control plurality of corn plants.
Reducing infield variability for corn yield per acre, evaluated by a lower standard deviation of corn mean yield across a locus compared to control corn plants.
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