Compositions and methods comprising high energy density soybean meal

Abstract

Soybean meal compositions comprising high energy content are provided. The soybean meal compositions can have true metabolizable energy of at least about 1300 kcal / kg dry matter; sucrose content of at least about 10.5% dry basis; protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis; protein content of at least about 62% dry basis; and / or oligosaccharide content of about 1% or less dry basis. Populations of high energy soybean seeds, methods of producing a high energy soybean meal composition by processing the populations of high energy soybean seeds, and feed, food, or beverage products produced from the high energy soybean meal compositions or soybean seed populations are also provided.

Description

[0001] Attorney Docket No.: B88552 1690 WO (00455)

[0002] COMPOSITIONS AND METHODS COMPRISING HIGH ENERGY DENSITY SOYBEAN MEAL

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 738,260, filed December 23, 2024, which is incorporated by referenced herein in its entirety.

[0005] FIELD OF THE INVENTION

[0006] The present disclosure relates to the field of agricultural biotechnology, and more specifically, soybean meal compositions.

[0007] BACKGROUND OF THE INVENTION

[0008] Due to increasing concern around the sustainability and nutritional aspects of the consumption of animal-sourced products, there is a growing consumer interest and demand in plant-based meal. Soybeans represent an attractive renewable source of nutrients and energy for use in foodstuffs. However, the energy content, protein content, or sucrose content of soybean meal generated from commodity soybeans can be significantly less than that of animal-sourced counterparts.

[0009] Energy density is a valuable trait for soybean meal. Soy compositions having high energy content (e.g., high protein content and high sucrose content) could offer important commercial, nutritional, and environmental advantages.

[0010] SUMMARY OF THE INVENTION

[0011] Soybean meal compositions comprising high energy content (e.g., high protein and high sucrose content) are provided. Populations of high energy soybean seeds are also provided. Further, methods of producing a high energy soybean meal composition by processing the populations of high energy soybean seeds, and feed, food, or beverage products produced from the high energy soybean meal compositions or soybean seed populations are also provided.

[0012] In one aspect, provided is a soybean meal composition comprising true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM).

[0013] In some embodiments, the soybean meal composition contains sucrose content of at least about 10.5% dry basis.Attorney Docket No.: B88552 1690 WO (00455)

[0014] In some embodiments, the soybean meal composition contains TMEn that is greater by at least about 10% relative to a control soybean meal composition derived from a control, e.g., commodity soybean cultivar.

[0015] In some embodiments, the soybean meal composition contains TMEn of at least about 1350 kcal / kg DM. In some embodiments, the soybean meal composition contains protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

[0016] In some embodiments, the soybean meal composition contains protein content of at least about 62% dry basis.

[0017] In some embodiments, the soybean meal composition contains oligosaccharide content of about 1% or less dry basis.

[0018] In some embodiments, the soybean meal contains soybean meal derived from: soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; or soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008; or an offspring plant or a parent plant of soybean cultivar 6073069 or 6167952.

[0019] In some embodiments, the soybean meal composition comprises a de-fatted soybean meal composition.

[0020] In one aspect, provided herein is a population of soybean seeds, comprising protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis.

[0021] In one aspect, provided herein is a population of soybean seeds, comprising oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis and sucrose content of at least about 8.25% dry basis.

[0022] In some embodiments, the population of soybean seeds has protein content of at least about 44% dry basis.

[0023] In some embodiments, the population of soybean seeds has oligosaccharide content of about 1% or less.

[0024] In some embodiments, the population of soybean seeds contains seeds of: soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008; or an offspring plant or a parent plant of the soybean cultivar 6073069or the soybean cultivar 6167952.

[0025] In some embodiments, provided is a soybean meal composition produced from the population of soybean seeds provided herein. In some embodiments, the soybean meal composition contains true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM). InAttorney Docket No.: B88552 1690 WO (00455)

[0026] some embodiments, the soybean meal composition contains TMEn that is greater by at least about 10% relative to a control soybean meal composition derived from a commodity soybean cultivar.

[0027] In some embodiments, the soybean meal composition contains protein content of at least about 62% dry basis; oligosaccharide content of about 1% or less dry basis; sucrose content of at least about 10.5% dry basis; and / or TMEn of at least about 1350 kcal / kg, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis. In some embodiments, the soybean meal composition comprises a de-fatted soybean meal.

[0028] In one aspect, provided herein is a method for producing a soybean meal composition. The method includes processing a population of soybean seeds to provide the soybean meal composition. The population of soybean seeds contains (i) protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis or (ii) oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis, and the soybean meal composition contains true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM) or TMEn that is greater by at least about 10% relative to a control soybean meal composition derived from a commodity soybean cultivar.

[0029] In some embodiments, the population of soybean seeds contains protein content of at least about 44% dry basis. In some embodiments, the population of soybean seeds contains oligosaccharide content of about 1% or less.

[0030] In some embodiments, the population of soybean seeds contains seeds of soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008; or an offspring plant or a parent plant of the soybean cultivar 6073069 or the soybean cultivar 6167952.

[0031] In some embodiments, the soybean meal composition contains TMEn of at least about 1350 kcal / kg DM, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

[0032] In some embodiments, the population of soybean seeds contains oligosaccharide content of about 1% or less.

[0033] In some embodiments, processing the population of soybean seeds contains fermenting, hexane extraction, drying, dehulling, grinding, heating, toasting, flaking, cracking, and / or defatting of the population of soybean seeds.

[0034] In some embodiments, provided herein is a soybean meal composition produced by the method provided herein.

[0035] In some embodiments, the soybean meal composition contains protein content of at least about 62% dry basis; oligosaccharide content of about 1% or less dry basis; sucrose content of atAttorney Docket No.: B88552 1690 WO (00455)

[0036] least about 10.5% dry basis; , and / or TMEn of at least about 1350 kcal / kg, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

[0037] In some embodiments, the soybean meal composition contains a de-fatted soybean meal composition.

[0038] In some embodiments, the soybean meal composition contains feed, protein isolate, protein concentrate, protein flakes, and / or textured soy protein (TSP).

[0039] In some embodiments, provided herein is a feed, food, or beverage product produced from the soybean composition or the population of soybean seeds provided herein. In some embodiments, the feed, food, or beverage product contains animal feed, aquatic animal feed, soy alternative meat products, soy alternative egg products, soy alternative dairy products, soy energy bars, soy food products, soy infant formula, or soy baby foods.

[0040] DETAILED DESCRIPTION OF THE INVENTION

[0041] The present disclosure now will be described more fully hereinafter. The disclosure may be embodied in many different forms and should not be construed as limited to the aspects set forth herein; rather, these aspects are provided so that this disclosure will satisfy applicable legal requirements.

[0042] I. Definitions

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0044] As used herein, “a,” “an,” or “the” can mean one or more than one. For example, “a” cell can mean a single cell or a multiplicity of cells. Further, the term “a plant” may include a plurality of plants.

[0045] As used herein, unless specifically indicated otherwise, the word “or” is used in the inclusive sense of “and / or” and not the exclusive sense of “either / or.”

[0046] The term “about” or “approximately” usually means within 5%, or more preferably within 1%, of a given value or range.

[0047] The terms “comprises,” “comprising,” “includes,” “including,” “having” and their conjugates mean “including but not limited to.”

[0048] Various embodiments of this disclosure may be presented in a range format. It should be noted that whenever a value or range of values of a parameter are recited, it is intended that values and ranges intermediate to the recited values are also part of this disclosure. It should be understoodAttorney Docket No.: B88552 1690 WO (00455)

[0049] that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1-10 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 1 to 6, from 1 to 7, from 1 to 8, from 1 to 9, from 2 to 4, from 2 to 6, from 2 to 8, from 2 to 10, from 3 to 6, etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. This applies regardless of the breadth of the range.

[0050] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases “ranging / ranges between” a first indicate number and a second indicate number and “ranging / ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals there between. The recitation of a numerical range for a variable is intended to convey that the present disclosure may be practiced with the variable equal to any of the values within that range. Thus, for a variable which is inherently discrete, the variable can be equal to any integer value within the numerical range, including the end-points of the range. Similarly, for a variable which is inherently continuous, the variable can be equal to any real value within the numerical range, including the end-points of the range. As an example, and without limitation, a variable which is described as having values between 0 and 2 can take the values 0, 1 or 2 if the variable is inherently discrete, and can take the values 0.0, 0.1, 0.01, 0.001, or any other real values =0 and =2 if the variable is inherently continuous.

[0051] A “plant” refers to a whole plant, any part thereof, or a cell or tissue culture derived from a plant, comprising any of: whole plants, plant components or organs (e.g., leaves, stems, roots, embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, pulp, juice, kernels, ears, cobs, husks, stalks, root tips, anthers, etc.), plant tissues, seeds, plant cells, protoplasts and / or progeny of the same. A plant cell is a biological cell of a plant, taken from a plant or derived through culture of a cell taken from a plant. Grain is intended to mean the mature seed produced by commercial growers for purposes other than growing or reproducing the species. Progeny, variants, and mutants of the regenerated plants are also included within the scope of the invention.

[0052] As used herein, a “soybean plant” refers to a plant of species Glycine max (L) and includes all plant varieties that can be bred with soybean, including wild soybean species such as Glycine soja.

[0053] “White flake protein” as used herein refers to a protein composition obtained by de-hulling, flaking, and defatting plants or plant parts (e.g., legume plants or plant parts) by solvent (e.g.,Attorney Docket No.: B88552 1690 WO (00455)

[0054] hexane) extraction, with limited use of heat to run off the solvent (Lusas and Riaz, 1995). White flake protein is an intermediate product in the production of plant protein concentrates and isolates. In contrast to conventional toasted plant meal (e.g., soybean meal), white flakes contains undenaturated proteins due to the very mild heat treatment. Thus, little or no reduction of protease inhibitors would be expected. The undenaturated proteins in white flakes may be advantageous in supporting binding properties during production of the extruded compound feed. White flakes can be used for human and animal consumption, including as a source of protein in aquaculture feeds for any type of fish or aquatic animal in a farmed or wild environment.

[0055] A “protein product” or “protein composition” refers to any protein composition or product isolated, extracted, and / or produced from plants or plant parts (e.g., seed) and includes isolates, concentrates, and flours, e.g., flake, white flake, soy / soybean protein composition, soy / soybean protein concentrate (SPC / PPC), soy / soybean protein isolate (SPI / PPI), soy / soybean flour, texturized vegetable protein (TVP), or textured soy / soybean protein (TSP / TPP)). Plant protein compositions of the present disclosure can be a concentrated protein solution (e.g., soybean protein concentrate solution) in which the protein is in a higher concentration than the protein in the plant from which the protein composition is derived. The protein composition can comprise multiple proteins as a result of the extraction or isolation process. In specific embodiments, the protein composition can further comprise stabilizers, excipients, drying agents, desiccating agents, anticaking agents, or any other ingredient to make the protein fit for the intended purpose. The protein composition can be a solid, liquid, gel, or aerosol and can be formulated as a powder. The protein composition can be extracted in a powder form from a plant and can be processed and produced in different ways, such as: (i) as an isolate - through the process of wet fractionation, which has the highest protein concentration; (ii) as a concentrate - through the process of dry fractionation, which are lower in protein concentration; and / or (Hi) in textured form - when it is used in food products as a substitute for other products, such as meat substitution (e.g. a “meat” patty). Protein isolate can be derived from defatted soy / soybean flour with a high solubility in water, as measured by the nitrogen solubility index (NSI). The aqueous extraction is carried out at a pH below 9. The extract is clarified to remove the insoluble material and the supernatant liquid is acidified to a pH range of 4-5. The precipitated protein-curd is collected and separated from the whey by centrifuge. The curd can be neutralized with alkali to form the sodium proteinate salt before drying. Protein concentrate can be produced by immobilizing the soy globulin proteins while allowing the soluble carbohydrates, whey proteins, and salts to be leached from the defatted flakes or flour. The protein is retained by one or more of several treatments: leaching with 20-80% aqueous alcohol / solvent, leaching with aqueous acids in the isoelectric zone of minimum protein solubility, pH 4-5; leaching with chilled water (which may involve calcium or magnesium cations), and leaching with hot waterAttorney Docket No.: B88552 1690 WO (00455)

[0056] of heat-treated defatted protein meal / flour (e.g., soy meal / flour). Any of the process provided herein can result in a product that is 70% protein, 20% carbohydrates (2.7 to 5% crude fiber), 6% ash and about 1% oil, but the solubility may differ. As an example, one ton (t) of defatted soybean flakes can yield about 750 kg of soybean protein concentrate.

[0057] “Soybean flour” or “soy flour” refers to defatted soybeans where special care was taken during desolventizing (not toasted) to minimize denaturation of the protein and to retain a high Nitrogen Solubility Index (NSI) in making the flour. Soy flour is the starting material for production of soy concentrate and soy protein isolate. Defatted soy flour is obtained from solvent extracted flakes, and contains less than 1% oil. Full-fat soy flour is made from unextracted, dehulled beans, and contains about 18% to 20% oil. Due to its high oil content, a specialized Alpine Fine Impact Mill must be used for grinding rather than the more common hammer mill. Low-fat soy flour is made by adding back some oil to defatted soy flour. The lipid content varies according to specifications, usually between 4.5% and 9%. High-fat soy flour can also be produced by adding back soybean oil to defatted flour at the level of 15%. Lecithinated soy flour is made by adding soybean lecithin to defatted, low-fat or high-fat soy flours to increase their dispersibility and impart emulsifying properties. The lecithin content varies up to 15%.

[0058] “Texturized vegetable protein” (TVP), “Textured vegetable protein,” which includes “textured soy protein” (TSP), soy meat, or soya chunks refers to a defatted plant (e.g., soy) flour product, a by-product of extracting plant (e.g., soybean) oil. It can be used as a meat analogue or meat extender. It is quick to cook, with a protein content comparable to certain meats. TVP can be produced from any protein-rich seed meal left over from vegetable oil production. A wide range of pulse seeds other than soybean, such as lentils, soybeans, and fava beans, or peanut may be used for TVP production. TVP can be made from high protein (e.g., 50%) soy isolate, flour, or concentrate, and can also be made from cottonseed, wheat, and oats. It is extruded into various shapes (chunks, flakes, nuggets, grains, and strips) and sizes, exiting the nozzle while still hot and expanding as it does so. The defatted thermoplastic proteins are heated to 150-200 °C, which denatures them into a fibrous, insoluble, porous network that can soak up as much as three times its weight in liquids. As the pressurized molten protein mixture exits the extruder, the sudden drop in pressure causes rapid expansion into a puffy solid that is then dried. As much as 50% protein when dry, TVP can be rehydrated at a 2: 1 ratio, which drops the percentage of protein to an approximation of ground meat at 16%. TVP can be used as a meat substitute. When cooked together, TVP can help retain more nutrients from the meat by absorbing juices normally lost. Also provided herein are methods of isolating, extracting, or preparing any of the protein compositions or protein products provided herein from plants or plant parts.Attorney Docket No.: B88552 1690 WO (00455)

[0059] An “oil product” or “oil composition” refers to any oil composition or product isolated, extracted, and / or produced from plants or plant parts (e.g., seed). Soybean oil have broad application in food products and industrial uses. Soybean oil products of the present disclosure include anti-static agents, caulking compounds, disinfectants, fungicides, inks, paints, protective coatings, wallboard, anti-foam agents, alcohol, margarine, paint, ink, rubber, shortening, cosmetics, and alkyd resins. Alkyd resins are dissolved in carrier solvents to make oil-based paints. The basic chemistry for converting vegetable oils into an alkyd resin under heat and pressure is well understood to those of skill in the art.

[0060] Plant cells possess nuclear, plastid, and mitochondrial genomes. Accordingly, by “chromosome” or “chromosomal” is intended the nuclear, plastid, or mitochondrial genomic DNA. “Genome” as it applies to plant cells encompasses not only chromosomal DNA found within the nucleus, but organelle DNA found within subcellular components (e.g., mitochondria or plastids) of the cell. The compositions and methods disclosed herein are not limited to mutations made in the genomic DNA of the plant nucleus, but may be used to modify the sequence of the nuclear, plastid, and / or mitochondrial genome, or to modulate the expression of a gene or genes encoded by the nuclear, plastid, and / or mitochondrial genome. In certain embodiments, a mutation is created in the genomic DNA of an organelle (e.g. a plastid and / or a mitochondrion). In certain embodiments, a mutation is created in extrachromosomal nucleic acids (including RNA) of the plant, cell, or organelle of a plant. Nonlimiting examples include creating mutations in supernumerary chromosomes (e.g. B chromosomes), plasmids, and / or vector constructs used to deliver nucleic acids to a plant. It is anticipated that new nucleic acid forms will be developed and yet fall within the scope of the claimed invention when used with the teachings described herein.

[0061] As used herein, the term “gene” or “coding sequence,” herein used interchangeably, refers to a functional nucleic acid unit encoding a protein, polypeptide, or peptide. As will be understood by those in the art, this functional term includes genomic sequences, cDNA sequences, and smaller engineered gene segments that express, or may be adapted to express proteins, polypeptides, domains, peptides, fusion proteins, and mutants. A gene may include a regulatory region, e.g., a promoter region or a 5 ’untranslated region, that regulates transcription or translation of the encoded gene.

[0062] As used herein, the term a “nucleic acid,” used interchangeably with a “nucleotide,” refers to a molecule consisting of a nucleoside and a phosphate that serves as a component of DNA or RNA. For instance, nucleic acids include adenine, guanine, cytosine, uracil, and thymine.

[0063] As used herein, “allele” refers to an alternative nucleic acid sequence at a particular locus. The length of an allele can be as small as one nucleotide base. For example, a first allele can occur on one chromosome, while a second allele occurs on a second homologous chromosome, e.g., asAttorney Docket No.: B88552 1690 WO (00455)

[0064] occurs for different chromosomes of a heterozygous individual, or between different homozygous or heterozygous individuals in a population. “Locus” as used herein refers to a chromosome region or chromosomal region where a polymorphic nucleic acid, trait determinant, gene, or marker is located.

[0065] As used herein, “function” of a gene, a peptide, a protein, or a molecule refers to activity of a gene, a peptide, a protein, or a molecule.

[0066] As used herein, a “mutation” is any change in a nucleic acid sequence. Nonlimiting examples comprise insertions, deletions, duplications, substitutions, inversions, and translocations of any nucleic acid sequence, regardless of how the mutation is brought about and regardless of how or whether the mutation alters the functions or interactions of the nucleic acid. For example and without limitation, a mutation may produce altered enzymatic activity of a ribozyme, altered base pairing between nucleic acids (e.g. RNA interference interactions, DNA-RNA binding, etc.), altered mRNA folding stability, and / or how a nucleic acid interacts with polypeptides (e.g. DNA-transcription factor interactions, RNA-ribosome interactions, gRNA-endonuclease reactions, etc.). A mutation might result in the production of proteins with altered amino acid sequences (e.g. missense mutations, nonsense mutations, frameshift mutations, etc.) and / or the production of proteins with the same amino acid sequence (e.g. silent mutations). Certain synonymous mutations may create no observed change in the plant while others that encode for an identical protein sequence nevertheless result in an altered plant phenotype (e.g. due to codon usage bias, altered secondary protein structures, etc.). Mutations may occur within coding regions (e.g., open reading frames) or outside of coding regions (e.g., within promoters, terminators, untranslated elements, or enhancers), and may affect, for example and without limitation, gene expression levels, gene expression profdes, protein sequences, and / or sequences encoding RNA elements such as tRNAs, ribozymes, ribosome components, and microRNAs.

[0067] As used herein with respect to a parameter, the term “increased” or “increasing” or “increase” refers to a detectable (e.g., at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, 120%, 150%, 200%, 300%, 400%, 500%, or more) positive change in the parameter from a comparison control, e.g., an established normal or reference level of the parameter, or an established standard control. Accordingly, the terms “increased,” “increase,” and the like encompass both a partial increase and a significant increase compared to a control.

[0068] As used herein with respect to a parameter, the term “decreased” or “decreasing” or “decrease” or “reduced” or “reducing” or “reduce” or “lower” or “loss” refers to a detectable (e.g., at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%) negative change in the parameter from aAttorney Docket No.: B88552 1690 WO (00455)

[0069] comparison control, e.g., an established normal or reference level of the parameter, or an established standard control. Accordingly, the terms “decreased,” “reduced,” and the like encompass both a partial reduction and a complete reduction compared to a control.

[0070] The term “isolated” refers to at least about partially separated from the natural environment e.g., from a plant cell.

[0071] As used herein, “fertilization” and / or “crossing” broadly includes bringing the genomes of gametes together to form zygotes but also broadly may include pollination, syngamy, fecundation and other processes related to sexual reproduction. Typically, a cross and / or fertilization occurs after pollen is transferred from one flower to another, but those of ordinary skill in the art will understand that plant breeders can leverage their understanding of fertilization and the overlapping steps of crossing, pollination, syngamy, and fecundation to circumvent certain steps of the plant life cycle and yet achieve equivalent outcomes, for example, a plant or cell of a soybean cultivar described herein. In certain embodiments, a user of this innovation can generate a plant of the claimed invention by removing a genome from its host gamete cell before syngamy and inserting it into the nucleus of another cell. While this variation avoids the unnecessary steps of pollination and syngamy and produces a cell that may not satisfy certain definitions of a zygote, the process falls within the definition of fertilization and / or crossing as used herein when performed in conjunction with these teachings. In certain embodiments, the gametes are not different cell types (i.e. egg vs. sperm), but rather the same type and techniques are used to effect the combination of their genomes into a regenerable cell. Other embodiments of fertilization and / or crossing include circumstances where the gametes originate from the same parent plant, i.e. a “self’ or “self-fertilization.” While selfing a plant does not require the transfer pollen from one plant to another, those of skill in the art will recognize that it nevertheless serves as an example of a cross,just as it serves as a type of fertilization. Thus, methods and compositions taught herein are not limited to certain techniques or steps that must be performed to create a plant or an offspring plant of the claimed invention, but rather include broadly any method that is substantially the same and / or results in compositions of the present disclosure.

[0072] As used herein, the term “method” refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

[0073] As used herein, the term “population” refers to a set comprising any number, including one, of individuals, objects, or data from which samples are taken for evaluation, e.g., estimating quantitative trait locus (QTL) effects. Most commonly, the terms relate to a breeding population ofAttorney Docket No.: B88552 1690 WO (00455)

[0074] plants from which members are selected and crossed to produce progeny in a breeding program. A population of plants can include the progeny of a single breeding cross or a plurality of breeding crosses and can be either actual plants or plant derived material, or in silico representations of plants. The member of a population need not be identical to the population members selected for use in subsequent cycles of analyses, nor does it need to be identical to those population members ultimately selected to obtain a final progeny of plants. Often, a plant population is derived from a single biparental cross but can also derive from two or more crosses between the same or different parents. Although a population of plants can comprise any number of individuals, those of skill in the art will recognize that plant breeders commonly use population sizes ranging from one or two hundred individuals to several thousand, and that the highest performing 5-20% of a population is what is commonly selected to be used in subsequent crosses in order to improve the performance of subsequent generations of the population in a plant breeding program.

[0075] As used herein, “quantitative trait locus” (QTL) or “quantitative trait loci” (QTLs) refer to a genetic domain that effects a phenotype that can be described in quantitative terms and can be assigned a “phenotypic value” which corresponds to a quantitative value for the phenotypic trait.

[0076] As used herein, the terms “phenotype,” or “phenotypic trait,” or “trait” refers to one or more detectable characteristics of a cell or organism which can be influenced by genotype. The phenotype can be observable to the naked eye, or by any other means of evaluation known in the art, e.g., microscopy, biochemical analysis, genomic analysis, an assay for a particular disease tolerance, etc. In some cases, a phenotype is directly controlled by a single gene or genetic locus, e.g., a “single gene trait.” In other cases, a phenotype is the result of several genes. In specific embodiments, the phenotype of soybean seeds is a high energy density phenotype.

[0077] As used herein, “allele” refers to an alternative nucleic acid sequence at a particular locus. The length of an allele can be as small as one nucleotide base. For example, a first allele can occur on one chromosome, while a second allele occurs on a second homologous chromosome, e.g., as occurs for different chromosomes of a heterozygous individual, or between different homozygous or heterozygous individuals in a population.

[0078] As used herein, “locus” is a chromosome region or chromosomal region where a polymorphic nucleic acid, trait determinant, gene, or marker is located. A locus may represent a single nucleotide, a few nucleotides or a large number of nucleotides in a genomic region. The loci of this disclosure comprise one or more polymorphisms in a population; e.g., alternative alleles are present in some individuals. A “gene locus” is a specific chromosome location in the genome of a species where a specific gene can be found.

[0079] An allele of a QTL can, as used herein, can comprise multiple genes or other genetic factors even within a contiguous genomic region or linkage group, such as a haplotype. As used herein, anAttorney Docket No.: B88552 1690 WO (00455)

[0080] allele of a QTL can therefore encompasses more than one gene or other genetic factor where each individual gene or genetic component is also capable of exhibiting allelic variation and where each gene or genetic factor is also capable of eliciting a phenotypic effect on the quantitative trait in question. In an embodiment of the present invention the allele of a QTL comprises one or more genes or other genetic factors that are also capable of exhibiting allelic variation. The use of the term “an allele of a QTL” is thus not intended to exclude a QTL that comprises more than one gene or other genetic factor. Specifically, an “allele of a QTL” in the present in the invention can denote a haplotype within a haplotype window wherein a phenotype can be disease resistance. A haplotype window is a contiguous genomic region that can be defined, and tracked, with a set of one or more polymorphic markers wherein said polymorphisms indicate identity by descent. A haplotype within that window can be defined by the unique fingerprint of alleles at each marker. As used herein, an allele is one of several alternative forms of a gene occupying a given locus on a chromosome. When all the alleles present at a given locus on a chromosome are the same, that plant is homozygous at that locus. If the alleles present at a given locus on a chromosome differ, that plant is heterozygous at that locus.

[0081] As used herein, a “haplotype” is the genotype of an individual at a plurality of genetic loci. Typically, the genetic loci described by a haplotype are physically and genetically linked, e.g., in the same chromosome interval. A haplotype can also refer to a combination of SNP alleles located within a single gene.

[0082] As used herein, “polymorphism” means the presence of one or more variations in a population. A polymorphism may manifest as a variation in the nucleotide sequence of a nucleic acid or as a variation in the amino acid sequence of a protein. Polymorphisms include the presence of one or more variations of a nucleic acid sequence or nucleic acid feature at one or more loci in a population of one or more individuals. The variation may comprise but is not limited to one or more nucleotide base changes, the insertion of one or more nucleotides or the deletion of one or more nucleotides. A polymorphism may arise from random processes in nucleic acid replication, through mutagenesis, as a result of mobile genomic elements, from copy number variation and during the process of meiosis, such as unequal crossing over, genome duplication and chromosome breaks and fusions. The variation can be commonly found or may exist at low frequency within a population, the former having greater utility in general plant breeding and the latter may be associated with rare but important phenotypic variation. Useful polymorphisms may include single nucleotide polymorphisms (SNPs), insertions or deletions in DNA sequence (Indels), simple sequence repeats of DNA sequence (SSRs), a restriction fragment length polymorphism, and a tag SNP. A genetic marker, a gene, a DNA-derived sequence, a RNA-derived sequence, a promoter, a 5' untranslated region of a gene, a 3' untranslated region of a gene, microRNA, siRNA, a toleranceAttorney Docket No.: B88552 1690 WO (00455)

[0083] locus, a satellite marker, a transgene, mRNA, ds mRNA, a transcriptional profile, and a methylation pattern may also comprise polymorphisms. In addition, the presence, absence, or variation in copy number of the preceding may comprise polymorphisms.

[0084] As used herein, “SNP” or “single nucleotide polymorphism” means a sequence variation that occurs when a single nucleotide (A, T, C, or G) in the genome sequence is altered or variable.

[0085] As used herein, “marker,” or “molecular marker,” or “marker locus” is a term used to denote a nucleic acid or amino acid sequence that is sufficiently unique to characterize a specific locus on the genome

[0086] As used herein, a centimorgan (“cM”) is a unit of measure of recombination frequency and genetic distance between two loci. One cM is equal to a 1% chance that a marker at one genetic locus will be separated from a marker at, a second locus due to crossing over in a single generation.

[0087] As used herein, “introgression” refers to the transmission of a desired allele of a genetic locus from one genetic background to another.

[0088] As used herein, the term “crop performance” is used synonymously with “plant performance” and refers to of how well a plant grows under a set of environmental conditions and cultivation practices. Crop performance can be measured by any metric a user associates with a crop’s productivity (e.g., yield), appearance and / or robustness (e.g., color, morphology, height, biomass, maturation rate, etc.), product quality (e.g., fiber lint percent, fiber quality, seed protein content, seed white flake protein content, seed carbohydrate content, etc.), cost of goods sold (e.g., the cost of creating a seed, plant, or plant product in a commercial, research, or industrial setting) and / or a plant’s tolerance to disease (e.g., a response associated with deliberate or spontaneous infection by a pathogen) and / or environmental stress (e.g., drought, flooding, low nitrogen or other soil nutrients, wind, hail, temperature, day length, etc.). Crop performance can also be measured by determining a crop’s commercial value and / or by determining the likelihood that a particular inbred, hybrid, or variety will become a commercial product, and / or by determining the likelihood that the offspring of an inbred, hybrid, or variety will become a commercial product. Crop performance can be a quantity (e.g., the volume or weight of seed or other plant product measured in liters or grams) or some other metric assigned to some aspect of a plant that can be represented on a scale (e.g., assigning a 1-10 value to a plant based on its disease tolerance).

[0089] A “microbe” will be understood to be a microorganism, i.e. a microscopic organism, which can be single celled or multicellular. Microorganisms are very diverse and include all the bacteria, archaea, protozoa, fungi, and algae, especially cells of plant pathogens and / or plant symbionts. Certain animals are also considered microbes, e.g. rotifers. In various embodiments, a microbe can be any of several different microscopic stages of a plant or animal. Microbes also include viruses,Attorney Docket No.: B88552 1690 WO (00455)

[0090] viroids, and prions, especially those which are pathogens or symbionts to crop plants. A “pathogen” as used herein refers to a microbe that causes disease or harmful effects on plant health.

[0091] A “fungus” includes any cell or tissue derived from a fungus, for example whole fungus, fungus components, organs, spores, hyphae, mycelium, and / or progeny of the same. A fungus cell is a biological cell of a fungus, taken from a fungus or derived through culture of a cell taken from a fungus.

[0092] A “pest” is any organism that can affect the performance of a plant in an undesirable way. Common pests include microbes, animals (e.g. insects and other herbivores), and / or plants (e.g. weeds). Thus, a pesticide is any substance that reduces the survivability and / or reproduction of a pest, e.g. fungicides, bactericides, insecticides, herbicides, and other toxins.

[0093] “Tolerance” or “improved tolerance” in a plant to disease conditions (e.g. growing in the presence of a pest) will be understood to mean an indication that the plant is less affected by the presence of pests and / or disease conditions with respect to yield, survivability and / or other relevant agronomic measures, compared to a less tolerant, more "susceptible" plant. Tolerance is a relative term, indicating that a "tolerant" plant survives and / or performs better in the presence of pests and / or disease conditions compared to other (less tolerant) plants (e.g., a different soybean cultivar) grown in similar circumstances. As used in the art, “tolerance” is sometimes used interchangeably with “resistance,” although resistance is sometimes used to indicate that a plant appears maximally tolerant to, or unaffected by, the presence of disease conditions. Plant breeders of ordinary skill in the art will appreciate that plant tolerance levels vary widely, often representing a spectrum of more-tolerant or less-tolerant phenotypes, and are thus trained to determine the relative tolerance of different plants, plant lines or plant families and recognize the phenotypic gradations of tolerance.

[0094] “Yield” as used herein is defined as the measurable produce of economic value from a crop. This may be defined in terms of quantity and / or quality. Yield is directly dependent on several factors, for example, the number and size of the organs, plant architecture (for example, the number of branches), seed production, leaf senescence and more. Root development, nutrient uptake, stress tolerance, photosynthetic carbon assimilation rates, and early vigor may also be important factors in determining yield. Optimizing the abovementioned factors may therefore contribute to increasing crop yield. Yield can be measured and expressed by any means known in the art. In specific embodiments, yield is measured by seed weight or volume in a given harvest area.

[0095] A plant, or its environment, can be contacted with a wide variety of “agriculture treatment agents.” As used herein, an “agriculture treatment agent,” or “treatment agent,” or “agent” can refer to any exogenously provided compound that can be brought into contact with a plant tissue (e.g. a seed) or its environment that affects a plant’s growth, development and / or performance, including agents that affect other organisms in the plant’s environment when those effects subsequently alterAttorney Docket No.: B88552 1690 WO (00455)

[0096] a plant’s performance, growth, and / or development (e.g. an insecticide that kills plant pathogens in the plant’s environment, thereby improving the ability of the plant to tolerate the insect's presence). Agriculture treatment agents also include a broad range of chemicals and / or biological substances that are applied to seeds, in which case they are commonly referred to as seed treatments and / or seed dressings. Seed treatments are commonly applied as either a dry formulation or a wet slurry or liquid formulation prior to planting and, as used herein, generally include any agriculture treatment agent including growth regulators, micronutrients, nitrogen-fixing microbes, and / or inoculants. Agriculture treatment agents include pesticides (e.g. fungicides, insecticides, bactericides, etc.) hormones (abscisic acids, auxins, cytokinins, gibberellins, etc.) herbicides (e.g. glyphosate, atrazine, 2,4-D, dicamba, etc.), nutrients (e.g. a plant fertilizer), and / or a broad range of biological agents, for example a seed treatment inoculant comprising a microbe that improves crop performance, e.g. by promoting germination and / or root development. In certain embodiments, the agriculture treatment agent acts extracellularly within the plant tissue, such as interacting with receptors on the outer cell surface. In some embodiments, the agriculture treatment agent enters cells within the plant tissue. In certain embodiments, the agriculture treatment agent remains on the surface of the plant and / or the soil near the plant. In certain embodiments, the agriculture treatment agent is contained within a liquid. Such liquids include, but are not limited to, solutions, suspensions, emulsions, and colloidal dispersions. In some embodiments, liquids described herein will be of an aqueous nature. However, in various embodiments, such aqueous liquids that comprise water can also comprise water insoluble components, can comprise an insoluble component that is made soluble in water by addition of a surfactant, or can comprise any combination of soluble components and surfactants. In certain embodiments, the application of the agriculture treatment agent is controlled by encapsulating the agent within a coating, or capsule (e.g. microencapsulation). In certain embodiments, the agriculture treatment agent comprises a nanoparticle and / or the application of the agriculture treatment agent comprises the use of nanotechnology.

[0097] As used herein, “selecting” or “selection” in the context of marker-assisted selection or breeding refer to the act of picking or choosing desired individuals, normally from a population, based on certain pre-determined criteria.

[0098] In certain embodiments, a user can combine the teachings herein with high-density molecular marker profiles spanning substantially the entire soybean genome to estimate the value of selecting certain candidates in a breeding program in a process commonly known as genomic selection.

[0099] The patent and scientific literature referred to herein establishes knowledge that is available to those of skill in the art. The issued US patents, allowed applications, published foreignAttorney Docket No.: B88552 1690 WO (00455)

[0100] applications, and references, including GenBank database sequences, which are cited herein are hereby incorporated by reference to the same extent as if each was specifically and individually indicated to be incorporated by reference.

[0101] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference herein in their entirety.

[0102] II. Soybean Meal Compositions and Soy Food and Beverage Products

[0103] A high energy density soybean meal composition is provided. A “soybean meal composition,” as used herein, refers to any composition derived from the soybean plants, plant parts, seeds, including any oil products, sugar products, fiber products, protein products (such as protein concentrate, protein isolate, flake, white flake protein, or other protein product), seed hulls, meal, or flour, for a food, feed, aqua, or industrial product, soybean extract (e.g., sweetener, antioxidants, alkaloids, etc.), soybean concentrate (e.g., whole soybean concentrate or soybean part concentrate), soybean powder (e.g., formulated powder, seed flour), soybean biomass (e.g., dried biomass, such as crushed and / or powdered biomass), grains, soybean protein composition, soybean oil composition, and food and beverage products containing soybean compositions (e.g., soybean parts, soybean extract, soybean concentrate, soybean powder, soybean protein, soybean oil, and soybean biomass) described herein. Soybean parts and soybean products provided herein can be intended for animal or human consumption.

[0104] A “high energy density” composition, product, or plant part (e.g., seed) as used herein refers to a composition, product, or plant part containing higher true metabolizable energy (TME) per unit (e.g., per kg of the composition, product, or plant part) relative to a control composition, product, or plant part. “True metabolizable energy (TME)” as used herein refers to energy available from food for an animal (such as birds and E j ™ 8.32 ( - 8.23

[0105]

[0106]

[0107]

[0108] cattle) after accounting for energy

[0109]

[0110] losses from metabolism (e.g., shedding of intestinal cells, excretion of endogenous urinary compounds). The TME values can be corrected for nitrogen retention and endogenous losses to prevent overestimation of metabolizable energy values and to get a more accurate estimate of the available energy for the animal to use. Nitrogen correction can be performed by subtracting the energy associated with the nitrogen excreted. Nitrogen-corrected true metabolizable energy (TMEn) can be measured using standard methodology, for example a precision-fed rooster assay. In the precision-fed assay, roosters are usually fasted for 24-26 h and then crop-intubated with approximately 30 g of a test ingredient, followed by being fasted for an additional 48 h during which time excreta are collected quantitatively. Excreta can be analyzed for TMEn according to standard methods, for example according to Parsons et al. 1982 Poultry Science 61:2241-2246,Attorney Docket No.: B88552 1690 WO (00455)

[0111] Parsons & Araba 2000 Poultry Science 79(8): 1127-31, or Baker et al, 2011 Poultry Science 90(2):390-5. For example, TMEn values can be calculated using the following equations:

[0112] Step 1 : Mixed TMEn (kcal / g) =

[0113] &

[0114] Step 2: TMEn (kcal / g) =

[0115]

[0116] Mixed TMEn value (TMEnm) can be initially calculated for the ground com and test ingredient mixture (1:1) (step 1) followed by calculation of TMEn. In the above equations, FEfed is the gross energy of the feed (kcal), EEfed and EEfasted are the excreta energy (kcal) of the fed and fasted roosters, respectively, Nfed and Nfasted are the amount of nitrogen (g) retained in the fed and fasted birds, respectively, FI is feed intake, TMEncis the TMEn of the added ground com, and 0.5 corrects for the 1:1 mixture of com and test ingredient.

[0117] A control composition, product, or plant part may have been derived from a reference or commodity plant or plant part that does not have high energy density phenotype. A high energy density composition, product, or plant part can have high content of protein, carbohydrate, and / or oil. In specific embodiments, a high energy density composition, product, or plant part has high protein content and high sucrose content.

[0118] A soybean meal composition provided herein can have TMEn of at least about 1300 kcal / kg dry matter (kg DM). For example, a soybean meal composition provided herein can have TMEn of at least about 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500 kcal / kg DM, about 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500 kcal / kg DM or more, or about 1300-1350, 1350-1400, 1400-1450, or 1450-1500 kcal / kg DM. On the other hand, a control or reference soybean meal composition may have TMEn of about 1200-1260 kcal / kg DM. The soybean meal composition provided herein can have TMEn that is greater by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more, by about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more, or by about 5-10%, 7-13%, or 10-15% relative to a control or reference soybean meal composition. The control or reference soybean meal composition may be soybean meal composition derived from commodity soybean seeds or reference soybean seeds. One of ordinary skill in the art can select an appropriate control or reference. TMEn in a soybean meal composition can be measured by standard methods for measuring TMEn in a feed or food composition, including rooster assays (e.g., a precision-fed rooster assay) as described herein.

[0119] The percent composition of a given component in a soybean or soybean compositions mayAttorney Docket No.: B88552 1690 WO (00455)

[0120] be described on an “as-is” basis or on a “dry -weight” basis. The percent composition of a given component may be converted between an “as-is” basis and a “dry-weight” basis using the following equation:

[0121] (protein content, “dry- weight” basis, %) = (protein content, “as-is” basis, %) / [100% - (moisture content, %)]

[0122] The word “protein” in the above equation may be interchanged with any other soybean component for which conversion between an “as-is” and a “dry-weight” basis is needed, including sucrose or oligosaccharide. The moisture content of a soybean or soybean product may be determined using any suitable methods or techniques known in in the art. For example, methods known in the art for analyzing the composition of soybeans may also be used to analyze the content of soybeans.

[0123] In some embodiments, the soybean meal composition comprises protein content of at least about 62% dry basis. For example, the high energy density soybean meal composition provided herein can have at least about 55%, 56%, 57%, 57.5%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, or 70%; about 55%, 56%, 57%, 57.5%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, or 70% or more; or about 50%-75%, 55%-75%, 60%-75%, 61%-75%, 62%-75%, 63%-75%, 64%-75%, 65%-75%, 66%-75%, 67%-75%, 68%-75%, 69%-75%, 50%-70%, 55%-70%, 60%-70%, 61%-70%, 62%-70%, 63%-70%, 64%-70%, 65%-70%, 66%-70%, 67%-70%, 68%-70%, 69%-70%, 50%-65%, 55%-65%, 60%-65%, 61%-65%, 62%-65%, 63%-65%, or 64%-65% soybean protein on a dry weight basis. Soybean meal compositions having protein content of greater than 62% dry basis can be referred to as “ultra-high protein” compositions. On the other hand, a control or reference soybean meal composition may have protein content of about 50-60% dry basis. In some embodiments, the high energy density soybean meal composition has at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more protein content per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained. Protein content in a soybean meal composition or soybean seeds can be measured by standard methods for measuring protein content in a sample, for example by protein extraction and quantitation (e.g., BCA protein assay, Lowry protein assay, Bradford protein assay), spectroscopy, near-infrared reflectance (NIR) (e.g., analyzing 700-2500 nm), and nuclear magnetic resonance spectrometry (NMR). Protein content can also be measured by the Dumas method, by combusting samples at a high temperature in the presence of high-purity oxygen, analyzing the gas from combustion for nitrogen content using aAttorney Docket No.: B88552 1690 WO (00455)

[0124] thermal conductivity detector, and calculating the amount of protein present in the sample using a conversion factor. The industry standard conversion factor for soybean is 6.25.

[0125] In certain embodiments, a soybean meal composition provided herein has sucrose content of at least about 10.5% dry basis. For example, a soybean meal composition provided herein can have at least about 10.5%, 11%, 11.5%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%; about 10.5%, 11%, 11.5%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, or more; or about 10.5-13%, 13-17%, or 17-20% soybean sucrose on a dry weight basis. Soybean meal compositions having sucrose content of greater than 10.5% dry basis can be referred to as “ultra-high sucrose” compositions. On the other hand, a control or reference soybean meal composition may have sucrose content of about 6-9.5% dry basis. In some embodiments, the high energy density soybean meal composition has at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more sucrose content per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained. Sucrose content in a soybean meal composition or soybean seeds can be measured by standard methods for measuring sucrose content in a sample, including spectroscopy (near infrared spectroscopy), refractometry, high performance liquid chromatography with refractive index detection (HPLC-RI), ion chromatography, size exclusion chromatography with refractive index detection (HPSEC-RI), liquid chromatography with mass spectrometry (LC-MS), derivatization and gas chromatography (GC), and high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD).

[0126] In some embodiments, the soybean meal composition comprises oligosaccharide content of about 1% or less dry basis. Soybean meal compositions having oligosaccharide content of about 1% or less can be referred to as “low oligosaccharide” compositions. Oligosaccharide content in a soybean meal composition can be measured by standard methods for measuring oligosaccharide content in a sample, including spectroscopy (near infrared spectroscopy), refractometry, HPLC-RI, ion chromatography, HPSEC-RI, LC-MS, derivatization and GC, and HPAEC-PAD.

[0127] The soybean meal composition provided herein is a high energy density soybean meal with high TMEn, which can contain high sucrose content in combination with high protein content.

[0128] As another example, the soybean meal composition can comprise protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis. Soybean meal compositions having protein content of at least about 57.5% dry basis and sucrose content of at least about 11.5% dry basis can be referred to as “CoVai” compositions. A CoVai product or composition can have TMEn of at least about 1350 kcal / DM and / or TMEn that is greater by at least about 10% relative to a control product or composition, e.g., derived from a commodity soybean cultivar.

[0129] For example, a soybean meal composition provided herein can contain protein content of atAttorney Docket No.: B88552 1690 WO (00455)

[0130] least about 57.5% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 57.5% dry basis and sucrose content of at least about 12% dry basis; protein content of at least about 57.5% dry basis and sucrose content of at least about 13% dry basis; protein content of at least about 57.5% dry basis and sucrose content of at least about 14% dry basis; protein content of at least about 57.5% dry basis and sucrose content of at least about 15% dry basis; protein content of at least about 58% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 59% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 60% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 61% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 62% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 63% dry basis and sucrose content of at least about 11.5% dry basis; protein content of at least about 64% dry basis and sucrose content of at least about 11.5% dry basis; or protein content of at least about 65% dry basis and sucrose content of at least about 11.5% dry basis.

[0131] The soybean meal composition provided herein can be derived from or produced from a population of high energy density soybean seeds, e.g., soybean seeds with high protein content and / or high sucrose content, such as those provided herein. High energy density soybean seeds can have protein content of at least about 39% dry basis. For example, the high energy density soybean seeds provided herein can have protein content of at least about 38%, 39%, 40%, 41%, 42%, 43%, 44%. 45%. 44%. 47%. 48%, 49%, or 50%; about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44< / 0,47o / o,48o / o,49o / o,or 50% or more; or about 38-50%, 38-50%, 38-49%, 38-48%, 38-47%, 38-46%, 38-45%, 38-44%, 38-43%, 38-42%, 38-41%, 38-40%, 38-39%, 39-50%, 39-50%, 39-49%, 39-48%, 39-47%, 39-46%, 39-45%, 39-44%, 39-43%, 39-42%, 39-41%, 39-40%, 40-50%, 40-50%, 40-49%, 40-48%, 40-47%, 40-46%, 40-45%, 40-44%, 40-43%, 40-42%, 40-41%, 41-50%, 41-50%, 41-49%, 41-48%, 41-47%, 41-46%, 41-45%, 41-44%, 41-43%, or 41-42% on a dry weight basis. On the other hand, control or reference soybean seeds may have protein content of about 25-40% dry basis.

[0132] High energy density soybean seeds can have sucrose content of at least 8.5% dry basis. For example, high energy density soybean seeds can have sucrose content of at least about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%; about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, or more; or about 7-11%, 7-10.5%, 7-10%, 7-9.5%, 7-9%, 7-8.5%, 7-8%, 7-7.5%, 7.5-11%, 7.5-10.5%, 7.5-10%, 7.5-9.5%, 7.5-9%, 7.5-8.5%, 7.5-8%, 8-11%, 8-10.5%, 8-10%, 8-9.5%, 8-9%, 8-8.5%, 8.5-11%, 8.5-10.5%, 8.5-10%, 8.5-9.5%, or 8.5-9% on a dry weight basis. On the other hand, control or reference soybean seeds may have sucrose content of about 3-6.5% dry basis.Attorney Docket No.: B88552 1690 WO (00455)

[0133] The population of high energy density soybean seeds provided herein can comprise high protein content in combination with high sucrose content. Without wishing to be bound by theory, a negative (inverse) correlation can be observed between the trait of high protein content and the trait of high sucrose content in soybean seeds. Thus, it is challenging to obtain a cultivar having both high protein content and high sucrose content, and the high energy density soybean cultivars and seeds provided herein having high protein and high sucrose contents are surprising. For example, the population of high energy density soybean seeds can have the protein content of at least about 39% dry basis, and sucrose content of at least about 8.5% dry basis. Soybean seeds having protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis can be referred to as “CoVal” soybean seeds and can produce CoVai soybean meal compositions. For example, a population of soybean seeds provided herein, from which a high density soybean meal composition can be produced, can contain protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 9% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 9.5% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 10% dry basis; protein content of at least about 40% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 41% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 42% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 43% dry basis and sucrose content of at least about 8.5% dry basis; or protein content of at least about 44% dry basis and sucrose content of at least about 8.5% dry basis.

[0134] High energy density soybean seeds can have oil content of at least 21.5% dry basis. For example, high energy density soybean seeds can have oil content of at least about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; or about 21-25%, 25-30%, 21.5-25%, 21.5-23%, 23-25%, 25-28%, or 28-30% on a dry weight basis. On the other hand, control or reference soybean seeds may have oil content of about 16-20% dry basis.

[0135] In certain embodiments, the population of high energy density soybean seeds provided herein comprises high oil content in combination with high sucrose content. Without wishing to be bound by theory, a negative (inverse) correlation can be observed between the trait of high protein content and the trait of high sucrose and / or oil content in soybean seeds. Thus, it is challenging to obtain a cultivar having high sucrose and oil content without reduced protein content, and the high energy density soybean cultivars and seeds provided herein, having high sucrose and oil content (e.g., without reduced protein content) are surprising. For example, the population of high energy density soybean seeds can have the oil content of at least about 21.5% dry basis, and sucroseAttorney Docket No.: B88552 1690 WO (00455)

[0136] content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, and sucrose content of at least about 8.5% dry basis; oil content of at least about 21.5% dry basis and sucrose content of at least about 9% dry basis; oil content of at least about 21.5% dry basis and sucrose content of at least about 9.5% dry basis; oil content of at least about 21.5% dry basis and sucrose content of at least about 10% dry basis; oil content of at least about 22% dry basis and sucrose content of at least about 8.25% dry basis; oil content of at least about 23% dry basis and sucrose content of at least about 8.25% dry basis; oil content of at least about 24% dry basis and sucrose content of at least about 8.25% dry basis; oil content of at least about 25% dry basis and sucrose content of at least about 8.25% dry basis; or oil content of at least about 26% dry basis and sucrose content of at least about 8.25% dry basis.

[0137] In certain embodiments, the population of high energy density soybean seeds provided herein comprises high oil content, high protein content, and high sucrose content. For example, the population of high energy density soybean seeds can have the oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 8.5% dry basis; oil content of at least about 21.5% dry basis protein content of at least 41%, and sucrose content of at least about 9% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 9.5% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 10% dry basis; oil content of at least about 22% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 23% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 24% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 25% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 26% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 42%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 43%, and sucrose content of at least about 8.25% dry basis; or oil content of at least about 21.5% dry basis, protein content of at least 44%, and sucrose content of at least about 8.25% dry basis.

[0138] For example, the soybean meal composition provided herein can be derived or produced at least about in part from soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; or soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008; and / or an offspring plant or a parent plant of the soybean cultivar 6073069 orAttorney Docket No.: B88552 1690 WO (00455)

[0139] the soybean cultivar 6167952. The soybean cultivars 6073069, 6167952 and their seeds are described further in this disclosure. Without wishing to be bound by theory, seeds from soybean cultivars 6073069 and 6167952 can be CoVai seeds and can produce CoVai soybean meal compositions.

[0140] High energy density (e.g., high protein content and / or high sucrose content) soybean plants and seeds, such as soybean cultivars 6073069, , 6167952, and their seeds, can be identified by genetic markers, as further described herein. Although the compositions of the high energy density soybean meal compositions will differ from the source soybeans as a result of the processing methods described herein, the resulting high energy density soybean meal compositions may contain detectable remnants of genetic material associated with the source soybeans having high energy density. In certain embodiments, the soybean products provided herein may contain genetic material tied to particular genetic markers (e.g., biomarkers associated with high protein content, high sucrose content, or low oligosaccharide content) which were present in the source soybeans, and may be similarly characterized by genetic analysis to identify the same markers. In some embodiments, the high energy density soybean meal compositions provided herein and obtained from soybeans as utilized herein may be characterized by genetic analysis similarly to analysis of high energy density soybeans. In some embodiments, any suitable techniques known in the art may be used to characterize the high energy density soybean meal compositions provided herein as comprising one or more genetic markers associated with high energy density in soybeans.

[0141] The high energy density soybean meal composition provided herein has high energy density (e.g., high protein content and / or high sucrose content), including relative to the soybean seeds from which the soybean meal composition was derived. For example, the high energy density soybean meal composition can have at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more TMEn per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained.

[0142] Also provided herein is a feed, food, or beverage product produced from the soybean composition or the population of soybean seeds provided herein. The high energy density soybean meal compositions provided herein may be used as ingredients in various feed, food, and beverage applications. With respect to certain applications, the high energy density soybean meal compositions provided herein may demonstrate comparable and, in certain circumstances, superior properties as compared to other soybean ingredients or soybean compositions in the market, and thus may be advantageously incorporated into specific food products over competitor soybean products. The superior properties may include, but are not limited to higher energy density, higher protein content, higher sucrose content, or lower oligosaccharide content.

[0143] Feed, food, or beverage products of the present disclosure can contain soybeanAttorney Docket No.: B88552 1690 WO (00455)

[0144] compositions, e.g., seed composition, protein composition, or oil composition of the present disclosure. Feed, food, or beverage products can be meant for animal or human consumption. Feed, food, or beverage products of the present disclosure can include animal feed, ready-to-drink beverages, or protein shake powders, shakes (e.g., protein shakes), shake powders, health drinks, alternative meat products (e.g., meatless burger patties, meatless sausages), alternative egg products (e.g., eggless mayo), alternative dairy products (e.g., non-dairy whipped toppings, non-dairy milk, non-dairy creamer, non-dairy milk shakes, non-diary ice cream, non-dairy yogurt, non-dairy cheese), energy bars (e.g., protein energy bars), infant formula, baby foods, cereals, baked goods, edamame, tofu, and tempeh.

[0145] Plant parts (e.g., seeds) and plant products (e.g., soybean meal, seed compositions, protein compositions, oil compositions, feed, food, or beverage products) as disclosed herein can be meant for consumption by agricultural animals or for use as feed in an agriculture or aquaculture system. In specific embodiments, plant parts and plant products include animal feed (e.g., roughages -forage, hay, silage; concentrates - cereal grains, soybean cake) intended for consumption by bovine, porcine, poultry, lambs, goats, or any other agricultural animal. In some embodiments, plant parts and plant products include aquaculture feed for any type of fish or aquatic animal in a farmed or wild environment including, without limitation, trout, carp, catfish, salmon, tilapia, crab, lobster, shrimp, oysters, clams, mussels, and scallops.

[0146] Feed, food, or beverage products produced from high energy density soybeans or high energy density soybean meal compositions may contain detectable remnants of genetic material associated with the source soybeans having high energy density. In certain embodiments, the soybean products provided herein may contain genetic material tied to particular genetic markers (e.g., biomarkers associated with high protein content, high sucrose content, or low oligosaccharide content) which were present in the source soybeans, and may be similarly characterized by genetic analysis to identify the same markers. In some embodiments, the feed, food, or beverage products provided herein and obtained from soybeans or soybean meal compositions as utilized herein may be characterized by genetic analysis similarly to analysis of high energy density soybeans. In some embodiments, any suitable techniques known in the art may be used to characterize feed, food, or beverage products derived from high energy density soybeans or high energy density soybean meal compositions provided herein as comprising one or more genetic markers associated with high energy density in soybeans.

[0147] III. Populations of High Energy Density Soybean Seeds

[0148] A population of soybean seeds having high energy density is provided. High energy density soybean seeds can have protein content of at least about 39% dry basis. For example, the highAttorney Docket No.: B88552 1690 WO (00455)

[0149] energy density soybean seeds provided herein can have average protein content of at least about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44%, 47%, 48%, 49%, or 50%; about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44%, 47%, 48%, 49%, or 50% or more; or about 38-50%, 38-50%, 38-49%, 38-48%, 38-47%, 38-46%, 38-45%, 38-44%, 38-43%, 38-42%, 38-41%, 38-40%, 38-39%, 39-50%, 39-50%, 39-49%, 39-48%, 39-47%, 39-46%, 39-45%, 39-44%, 39-43%, 39-42%, 39-41%, 39-40%, 40-50%, 40-50%, 40-49%, 40-48%, 40-47%, 40-46%, 40-45%, 40-44%, 40-43%, 40-42%, 40-41%, 41-50%, 41-50%, 41-49%, 41-48%, 41-47%, 41-46%, 41-45%, 41-44%, 41-43%, or 41-42% on a dry weight basis. On the other hand, control or reference soybean seeds may have average protein content of about 25-40% dry basis.

[0150] In further embodiments, at least about a portion of the high energy density soybean seeds provided herein have protein content of at least about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44%. 47%. 48%, 49%, or 50%; about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44%, 47%, 48%, 49%, or 50% or more; or about 38-50%, 38-50%, 38-49%, 38-48%, 38-47%, 38-46%, 38-45%, 38-44%, 38-43%, 38-42%, 38-41%, 38-40%, 38-39%, 39-50%, 39-50%, 39-49%, 39-48%, 39-47%, 39-46%, 39-45%, 39-44%, 39-43%, 39-42%, 39-41%, 39-40%, 40-50%, 40-50%, 40-49%, 40-48%, 40-47%, 40-46%, 40-45%, 40-44%, 40-43%, 40-42%, 40-41%, 41-50%, 41-50%, 41-49%, 41-48%, 41-47%, 41-46%, 41-45%, 41-44%, 41-43%, or 41-42% on a dry weight basis. On the other hand, at least about a portion of control or reference soybean seeds may have protein content of about 25-40% dry basis.

[0151] In further embodiments, the majority of the high energy density soybean seeds provided herein have protein content of at least about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44%, 47%, 48%, 49%, or 50%; about 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 44%, 47%, 48%, 49%, or 50% or more; or about 38-50%, 38-50%, 38-49%, 38-48%, 38-47%, 38-46%, 38-45%, 38-44%, 38-43%, 38-42%, 38-41%, 38-40%, 38-39%, 39-50%, 39-50%, 39-49%, 39-48%, 39-47%, 39-46%, 39-45%, 39-44%, 39-43%, 39-42%, 39-41%, 39-40%, 40-50%, 40-50%, 40-49%, 40-48%, 40-47%, 40-46%, 40-45%, 40-44%, 40-43%, 40-42%, 40-41%, 41-50%, 41-50%, 41-49%, 41-48%, 41-47%, 41-46%, 41-45%, 41-44%, 41-43%, or 41-42% on a dry weight basis. On the other hand, the majority of control or reference soybean seeds may have protein content of about 25-40% dry basis.

[0152] High energy density soybean seeds can have average sucrose content of at least about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%; about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, or more; or about 7-11%, 7-10.5%, 7-10%, 7-9.5%, 7-9%, 7-8.5%, 7-8%, 7-7.5%, 7.5-11%, 7.5-10.5%, 7.5-10%, 7.5-9.5%, 7.5-9%, 7.5-8.5%, 7.5-8%, 8-11%, 8-10.5%, 8-10%, 8-9.5%, 8-9%, 8-8.5%, 8.5-11%, 8.5-10.5%, 8.5-10%, 8.5-9.5%, or 8.5-9% on a dry weight basis. On the other hand, control or reference soybean seeds may have average sucrose content ofAttorney Docket No.: B88552 1690 WO (00455)

[0153] about 3-6.5% dry basis.

[0154] In further embodiments, at least about a portion of high energy density soybean seeds have sucrose content of at least about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%; about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, or more; or about 7-11%, 7-10.5%, 7- 10%, 7-9.5%, 7-9%, 7-8.5%, 7-8%, 7-7.5%, 7.5-11%, 7.5-10.5%, 7.5-10%, 7.5-9.5%, 7.5-9%, 7.5- 8.5%, 7.5-8%, 8-11%, 8-10.5%, 8-10%, 8-9.5%, 8-9%, 8-8.5%, 8.5-11%, 8.5-10.5%, 8.5-10%, 8.5- 9.5%, or 8.5-9% on a dry weight basis. On the other hand, at least about a portion of the control or reference soybean seeds may have sucrose content of about 3-6.5% dry basis.

[0155] In further embodiments, the majority of high energy density soybean seeds have sucrose content of at least about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%; about 7%, 7.5%, 8%, 8.25%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, or more; or about 7-11%, 7-10.5%, 7-10%, 7-9.5%, 7-9%, 7-8.5%, 7-8%, 7-7.5%, 7.5-11%, 7.5-10.5%, 7.5-10%, 7.5-9.5%, 7.5-9%, 7.5-8.5%, 7.5-8%, 8-11%, 8-10.5%, 8-10%, 8-9.5%, 8-9%, 8-8.5%, 8.5-11%, 8.5-10.5%, 8.5-10%, 8.5-9.5%, or 8.5-9% on a dry weight basis. On the other hand, the majority of the control or reference soybean seeds may have sucrose content of about 3-6.5% dry basis.

[0156] High energy density soybean seeds can have average oil content of at least 21.5% dry basis. For example, high energy density soybean seeds can have oil content of at least about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; or about 21-25%, 25-30%, 21.5-25%, 21.5-23%, 23- 25%, 25-28%, or 28-30% on a dry weight basis. On the other hand, control or reference soybean seeds may have average oil content of about 16-20% dry basis.

[0157] In further embodiments, at least about a portion of high energy density soybean seeds have oil content of at least 21.5% dry basis. For example, high energy density soybean seeds can have oil content of at least about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; or about 21-25%.

[0158] 25-30%, 21.5-25%, 21.5-23%, 23-25%, 25-28%, or 28-30% on a dry weight basis. On the other hand, at least about a portion of the control or reference soybean seeds may have average oil content of about 16-20% dry basis.

[0159] In further embodiments, the majority of high energy density soybean seeds have oil content of at least 21.5% dry basis. For example, the majority of high energy density soybean seeds can have oil content of at least about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more; about 21%, 21.5%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, ormore; or about 21-25%, 25-30%, 21.5-25%, 21.5-23%, 23-25%, 25-28%, or 28-30% on a dry weight basis. On the other hand, the majority of the control or reference soybean seeds may have average oil content of about 16-20% dry basis.Attorney Docket No.: B88552 1690 WO (00455)

[0160] Content of a composition (e.g., protein content, sucrose content, oil content, oligosaccharide content, e.g., X% dry basis) in a population of soybean seeds as used herein may refer to average content (e.g., average protein content) of X% dry basis, at least a portion of the soybean seeds having the content (e.g., protein content) of X% dry basis, or the majority of the soybean seeds having the content (e.g., protein content) of X% dry basis.

[0161] The population of high energy density soybean seeds provided herein can comprise high protein content in combination with high sucrose content. Without wishing to be bound by theory, a negative (inverse) correlation can be observed between the trait of high protein content and the trait of high sucrose content in soybean seeds. Thus, it is challenging to obtain a cultivar having both high protein content and high sucrose content, and the high energy density soybean cultivars and seeds provided herein having high protein and high sucrose contents are surprising. In specific embodiments, the population of soybean seeds can have protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis. For example, a population of high energy density soybean seeds can have the protein content of at least about 39% dry basis, and sucrose content of at least about 8.5% dry basis, and can be referred to as “CoVal” soybean seeds, capable of producing CoVal soybean meal compositions. A population of high energy density soybean seeds can have the oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.5% dry basis, and can be referred to as “CoVal” soybean seeds, capable of producing CoVal soybean meal compositions.

[0162] For example, a population of soybean seeds provided herein can contain protein content of protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 9% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 9.5% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 10% dry basis; protein content of at least about 40% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 41% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 42% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 43% dry basis and sucrose content of at least about 8.5% dry basis; or protein content of at least about 44% dry basis and sucrose content of at least about 8.5% dry basis.

[0163] In certain embodiments, the population of high energy density soybean seeds provided herein has high oil content and high sucrose content. Without wishing to be bound by theory, a negative (inverse) correlation can be observed between the trait of high protein content and the trait of high sucrose and / or oil content in soybean seeds. Thus, it is challenging to obtain a cultivar having high sucrose and oil content without reduced protein content or in combination with highAttorney Docket No.: B88552 1690 WO (00455)

[0164] protein content. The high energy density soybean cultivars and seeds provided herein, having high sucrose and oil content (e.g., without reduced protein content) or having high sucrose, oil, and protein content are surprising. For example, the population of high energy density soybean seeds can have the oil content of at least about 21.5% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, and sucrose content of at least about 8.5% dry basis; oil content of at least about 21.5% dry basis and sucrose content of at least about 9% dry basis; oil content of at least about 21.5% dry basis and sucrose content of at least about 9.5% dry basis; oil content of at least about 21.5% dry basis and sucrose content of at least about 10% dry basis; oil content of at least about 22% dry basis and sucrose content of at least about 8.25% dry basis; oil content of at least about 23% dry basis and sucrose content of at least about 8.25% dry basis; oil content of at least about 24% dry basis and sucrose content of at least about 8.25% dry basis; oil content of at least about 25% dry basis and sucrose content of at least about 8.25% dry basis; or oil content of at least about 26% dry basis and sucrose content of at least about 8.25% dry basis.

[0165] In certain embodiments, the population of high energy density soybean seeds provided herein comprises high oil content, high protein content, and high sucrose content. For example, the population of high energy density soybean seeds can have the oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 8.5% dry basis; oil content of at least about 21.5% dry basis protein content of at least 41%, and sucrose content of at least about 9% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 9.5% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 41%, and sucrose content of at least about 10% dry basis; oil content of at least about 22% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 23% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 24% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 25% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 26% dry basis, protein content of at least 41%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 42%, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least 43%, and sucrose content of at least about 8.25% dry basis; or oil content of at least about 21.5% dry basis, protein content of at least 44%, and sucrose content of at least about 8.25% dry basis.

[0166] In some embodiments, the population of soybean seeds comprises oligosaccharide contentAttorney Docket No.: B88552 1690 WO (00455)

[0167] of about 1% or less, such as about 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, 0.1% or less, about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, about 1-0.7%, 0.7%-0.4%, or 0.4-0.1%. On the other hand, control or reference soybean seeds may have oligosaccharide content of greater than 1%.

[0168] High energy density seeds of the present disclosure include a representative sample of seeds from a plant of the present disclosure. In some embodiments, the population of soybean seeds comprises seeds of: soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008; and / or an offspring plant or a parent plant of the soybean cultivar 6073069 or the soybean cultivar 6167952. In certain embodiments, soybean cultivar 6073069 produces soybean seeds having oil content of at least about 21.5%, protein content of at least about 49%, and sucrose content of at least about 8.25%. In embodiments, soybean cultivar 6167952 produces soybean seeds having protein content of at least about 39% and sucrose content of at least about 8.5%. Soybean cultivar 6073069 and 6167952 may be CoVai varieties.

[0169] A deposit of proprietary soybean cultivars 6073069 and 6167952, disclosed herein and recited in the appended claims is maintained by the Applicant. A deposit of soybean cultivars 6073069 and 6167952 was made on December 12, 2024 with Provasoli-Guillard National Center for Marine Algae and Microbiota, Bigelow Laboratory for Ocean Sciences (NCMA), 60 Bigelow Drive, East Boothbay, Maine 04544. Access to this deposit will be available during the pendency of this application to persons determined by the Commissioner of Patents and Trademarks to be entitled thereto under 37 C.F.R. 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictions on the availability to the public of the variety will be irrevocably removed by affording access to a deposit of at least about 625 seeds of the same variety with NCMA. The deposit will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary, during that period.

[0170] To satisfy the enablement requirements of 35 U.S.C. § 112, and to certify that the deposit of the present disclosure meets the criteria set forth in 37 CFR § 1.801-1.809 and Manual of Patent Examining Procedure (MPEP) § 2402-2411.05, Applicant hereby makes the following statements regarding the deposited soybean cultivars 6073069 (deposited as NCMA No. 202412007) and 6167952 (deposited as NCMA No. 202412008):

[0171] If the deposit is made under the terms of the Budapest Treaty, the instant disclosure will be irrevocably and without restriction released to the public upon the granting of a patent.

[0172] If the deposit is made not under the terms of the Budapest Treaty, Applicant(s) providesAttorney Docket No.: B88552 1690 WO (00455)

[0173] assurance of compliance by following statements:

[0174] 1. During the pendency of this application, access to the disclosure will be afforded to the Commissioner upon request;

[0175] 2. All restrictions on availability to the public will be irrevocably removed upon granting of the patent under conditions specified in 37 CFR § 1.808;

[0176] 3. The deposit will be maintained in a public repository for a period of 30 years or 5 years after the last request or for the effective life of the patent, whichever is longer;

[0177] 4. A test of the viability of the biological material at the time of deposit will be conducted by the public depository under 37 CFR § 1.807; and

[0178] 5. The deposit will be replaced if it should ever become unavailable.

[0179] Access to this deposit will be available during the pendency of this application to persons determined by the Commissioner of Patents and Trademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 122. Upon granting of any claims in this application, all restrictions on the availability to the public of the variety will be irrevocably removed by affording access to a deposit with the NCMA.

[0180] In some embodiments, the population of high energy density soybean seeds provided herein comprise soybean seeds of soybean cultivar 6073069 or 6167952, or of their offspring or parent plants. Seeds from soybean cultivars 6073069 or 6167952, or their offspring or parent plant can be CoVai seeds and can produce CoVai soybean meal compositions.

[0181] The soybean plants or plant parts described above having high energy density (e.g., high protein content, high sucrose content, low oligosaccharide content) may be characterized by any suitable methods known in the art for any phenotypic characteristics. Additionally or alternatively, soybean plants or plant parts described herein having high energy density (e.g., high protein content, high sucrose content, low oligosaccharide content) may be characterized by genetic analysis, as having one or more genetic markers associated with the high energy density (or high protein content, high sucrose content, low oligosaccharide content) using any suitable techniques for detecting genetic markers.

[0182] In certain embodiments, a user can combine the teachings herein with molecular marker profiles spanning substantially the entire soybean genome to estimate the value of selecting certain candidates in a breeding program in a process commonly known as genomic selection.

[0183] In some embodiments, the population of soybeans used have higher energy density (e.g., higher TMEn), higher protein content, higher sucrose content, and / or lower oligosaccharide content as compared to a population of control soybeans, reference soybeans, or commodity soybeans. In some embodiments, the population of soybean provided herein, or used to produce high energy density soybean meal provided herein, have higher energy density, higher protein content, higherAttorney Docket No.: B88552 1690 WO (00455)

[0184] sucrose content, and / or lower oligosaccharide content as compared to a population of control, reference, or commodity soybeans, while having characteristics that are otherwise similar to those of a population of control, reference, or commodity soybeans. For example, the population of soybeans provided herein can have higher energy density, higher protein content, higher sucrose content, and / or lower oligosaccharide content as compared to a population of control, reference, or commodity soybeans, while having a similar average fat content, total dietary fiber content, and ash content as the population of control, reference, or commodity soybeans.

[0185] In some embodiments, the high energy density soybeans having high protein content, high sucrose content, and / or low oligosaccharide content provided herein are distinguished from commodity soybeans. Similarly, high energy density soybean meal compositions obtained from the soybeans having high energy density, high protein content, high sucrose content, and / or low oligosaccharide content described herein can be distinguished from soybean meal compositions obtained from commodity soybeans. Soybean meal compositions can include feed, soybean flour, soybean flakes, textured soy protein, soybean protein concentrates, and soybean protein isolates.

[0186] The present disclosure is not limited to whether the soybeans comprise transgenic polynucleotides or proteins. The soybeans used in the present Examples herein are non-transgenic and there are circumstances when using soybeans lacking transgenic traits, genome edits, or any other form of mutation (e.g., a change in a polynucleotide sequence) is necessary and / or beneficial. However, combining the disclosure herein with a wide range of transgenic plants, or plants containing genome edits or any other form of mutation to confer new traits or combinations thereof is also envisioned.

[0187] In certain embodiments, soybeans as disclosed herein may be obtained from soybean plants modified to exhibit at least about one desired trait, and / or combinations thereof. The present disclosures are not limited to any set of traits that can be considered desirable, but nonlimiting examples include male sterility, herbicide tolerance, pest tolerance, disease

[0188] tolerance, modified fatty acid metabolism, modified carbohydrate metabolism, modified seed yield, modified seed oil, modified seed protein, modified lodging resistance, modified shattering, modified iron-deficiency chlorosis, modified water use efficiency, and / or combinations thereof.

[0189] Also provided is a soybean meal composition produced from the population of soybean seeds provided herein. The soybean meal composition provided herein can be high density soybean meal composition, having at least about one characteristics described herein. For example, the soybean meal composition produced from the population of soybean seeds provided herein can contain protein content of at least about 62% dry basis; oligosaccharide content of about 1% or less dry basis; sucrose content of at least about 10.5% dry basis; ; and / or protein content of at least about 57.5% dry basis and sucrose content of at least about 11.5% dry basis. Other characteristicsAttorney Docket No.: B88552 1690 WO (00455)

[0190] of a soybean meal composition produced from the population of soybean seeds are described elsewhere herein. The soybean meal composition produced from the population of soybean seeds provided herein can comprise a de-fatted soybean meal composition. The soybean meal composition produced from the population of soybean seeds provided herein can be feed, protein isolate, protein concentrate, soybean flour, protein flakes, and / or textured soy protein (TSP).

[0191] For animal and human consumption, high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 can be used to produce edible protein ingredients which offer a healthier, less expensive replacement for animal protein in meats, as well as in dairy-type products. The soybeans produced by high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 can be processed to produce a texture and appearance similar to many other foods. For example, soybeans are the primary ingredient in many dairy product substitutes (e.g., soy milk, margarine, soy ice cream, soy yogurt, soy cheese, and soy cream cheese) and meat substitutes (e.g., veggie burgers). These substitutes are readily available in most supermarkets. Although soy milk does not naturally contain significant amounts of digestible calcium (the high calcium content of soybeans is bound to the insoluble constituents and remains in the soy pulp), many manufacturers of soy milk sell calcium-enriched products as well. Soy is also used in tempeh, where the beans (sometimes mixed with grain) are fermented into a solid cake.

[0192] Additionally, high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 can be used to produce various types of “fillers” in meat and poultry products. Food service, retail, and institutional (primarily school lunch and correctional) facilities regularly use such “extended” products, that is, products which contain soy fillers. Extension may result in diminished flavor, but fat and cholesterol are reduced by adding soy fillers to certain products. Vitamin and mineral fortification can be used to make soy products nutritionally equivalent to animal protein; the protein quality is already roughly equivalent.

[0193] In certain embodiments, high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 can be used to achieve certain grain content profiles in combination with another soybean cultivar or variety as part of a mixture of seeds or grain. In certain aspects, a mixture of harvested grain comprises two or more varieties blended together in a certain ratio whose combined content achieves certain threshold levels of a characteristic desirable to the user. For example, a user of high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 can combine the seed of high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 with the seed of another variety at a certain ratio into a container (e.g. a bag of seed, a compartment on a planting machine, etc.) and the two varieties can be planted and grown together in the same growing location (e.g. a commercial growing area) such that after the grain has been harvested from that growing locating the harvested grain product comprises a certain mixtureAttorney Docket No.: B88552 1690 WO (00455)

[0194] of the two varieties. In certain embodiments, mixing the at least about two varieties this way achieves a combined harvested grain composition with certain amounts of quality traits the user desires within the grain, for example, a certain percent content of protein, oil, carbohydrates or some other aspect of the grain, or a combination of thresholds thereof. In certain embodiments, harvesting a mixture of the at least about two varieties achieve a certain threshold of yield, e.g. mass of grain collected per acre.

[0195] Seed or grain of high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 may be used to achieve certain grain content profdes, such as protein, oil, carbohydrates or some other aspect of the grain, or a combination of thresholds thereof, in combination with another soybean cultivar or variety as part of a mixture of seeds or grain. In certain aspects, a mixture of harvested grain comprises two or more varieties blended together in a certain ratio whose combined content achieves certain threshold levels of a characteristic desirable to the user. For example, a user of high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 can combine the seed of high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 with the seed of another variety at a certain ratio into a container (e.g. a bag of seed, a compartment on a planting machine, etc.) and the two varieties can be planted and grown together in the same growing location (e.g. a commercial growing area) such that after the grain has been harvested from that growing locating the harvested grain product comprises a certain mixture of the two varieties. In certain embodiments, mixing the at least about two varieties this way achieves a combined harvested grain composition with certain amounts of quality traits the user desires within the grain, for example, a certain percent content of protein, oil, carbohydrates or some other aspect of the grain, or a combination of thresholds thereof. In certain embodiments, harvesting a mixture of the at least about two varieties achieves a certain threshold of yield, e.g. mass of grain collected per acre.

[0196] The mixing of the grain to achieve the desired mixture is performed after harvest, e.g. the seed of the two varieties are not mixed together and / or the two varieties are not harvested together as a mixture, but instead the grain of a first variety is collected and then combined with the grain collected from a second or additional varieties.

[0197] Seed or grain of high energy density soybean cultivars, such as soybean cultivar 6073069 or 6167952 may be used in combination with another cultivar or variety in a mixture of grain that achieves certain threshold levels of contents desired by a soybean processor or crush facility that facilitates the production of a processed soy composition like soybean meal, white flake, oil, protein extract, protein isolate, food, livestock feed, a raw material for industry, etc., including those described elsewhere herein.

[0198] Seed or grain of high energy density soybean cultivars, such as soybean cultivar 6073069Attorney Docket No.: B88552 1690 WO (00455)

[0199] or 6167952 may be used in combination with another cultivar or variety in a mixture of grain that achieves certain threshold levels of contents desired by a soybean processor or crush facility that facilitates the production of a processed soy composition like soybean meal, white flake, oil, protein extract, protein isolate, food, livestock feed, a raw material for industry, etc., including those described elsewhere herein.

[0200] IV. Breeding of High Energy Density Soybean Plants and Seeds

[0201] The population of high energy density soybean seeds of the present disclosure may be obtained by breeding soybean plants or seeds. The complexity of inheritance influences choice of the breeding method. Backcross breeding is used to transfer one or a few favorable genes for a highly heritable trait into a desirable cultivar. This approach has been used extensively for breeding cultivars with traits of interest. Various recurrent selection techniques are used to improve quantitatively inherited traits controlled by numerous genes. The use of recurrent selection in selfpollinating crops depends on the ease of pollination, the frequency of successful hybrids from each pollination, and the number of hybrid offspring from each successful cross.

[0202] Promising advanced breeding lines are thoroughly tested and compared to appropriate standards in environments representative of the commercial target area(s) for three or more years. The best lines are candidates for new commercial cultivars; those still deficient in a few traits may be used as parents to produce new populations for further selection.

[0203] A population refers to a set comprising any number, including one, of individuals, objects, or data from which samples are taken for evaluation, e.g. estimating QTL effects and / or disease tolerance. Most commonly, the terms relate to a breeding population of plants from which members are selected and crossed to produce progeny in a breeding program. A population of plants can include the progeny of a single breeding cross or a plurality of breeding crosses and can be either actual plants or plant derived material, or in silico representations of plants. The member of a population need not be identical to the population members selected for use in subsequent cycles of analyses nor does it need to be identical to those population members ultimately selected to obtain a final progeny of plants. Often, a plant population is derived from a single biparental cross but can also derive from two or more crosses between the same or different parents. Although a population of plants can comprise any number of individuals, those of skill in the art will recognize that plant breeders commonly use population sizes ranging from one or two hundred individuals to several thousand, and that the highest performing 5% to 20% of a population is what is commonly selected to be used in subsequent crosses in order to improve the performance of subsequent generations of the population in a plant breeding program.

[0204] These processes, which lead to the final step of marketing and distribution, usually takeAttorney Docket No.: B88552 1690 WO (00455)

[0205] from eight to twelve years from the time the first cross is made. Therefore, development of new cultivars is a time-consuming process that requires precise forward planning, efficient use of resources, and a minimum of changes in direction.

[0206] A most difficult task is the identification of individuals that are genetically superior, because for most traits the true genotypic value is masked by other confounding plant traits or environmental factors. One method of identifying a superior plant is to observe its performance relative to other experimental plants and to a widely grown standard cultivar. If a single observation is inconclusive, replicated observations provide a better estimate of its genetic worth.

[0207] The goal of soybean plant breeding is to develop new and superior soybean cultivars and hybrids. The breeder initially selects and crosses two or more parental lines, followed by repeated selfing and selection, producing many new genetic combinations. The breeder can theoretically generate billions of different genetic combinations via crossing, selection, selfing and mutations. Therefore, a breeder will never develop the same line, or even very similar lines, having the same soybean traits from the exact same parents.

[0208] Each year, the plant breeder selects the germplasm to advance to the next generation. This germplasm is grown under different geographical climate and soil conditions and further selections are then made during and at the end of the growing season. The cultivars that are developed are unpredictable because the breeder's selection occurs in environments with no control at the DNA level, and with millions of different possible genetic combinations being generated. A breeder of ordinary skill in the art cannot predict the final resulting lines he develops, except possibly in a very gross and general fashion. The same breeder cannot produce the same cultivar twice by using the same original parents and the same selection techniques. This unpredictability results in the expenditure of large amounts of research monies to develop superior new soybean cultivars.

[0209] The development of new soybean cultivars requires the development and selection of soybean varieties, the crossing of these varieties and selection of superior hybrid crosses. The hybrid seed is produced by manual crosses between selected male-fertile parents or by using male sterility systems. These hybrids are selected for certain single gene traits such as pod color, flower color, pubescence color or herbicide resistance which indicate that the seed is truly a hybrid.

[0210] Additional data on parental lines, as well as the phenotype of the hybrid, influence the breeder's decision whether to continue with the specific hybrid cross.

[0211] Breeding programs combine desirable traits from two or more cultivars or various broad-based sources into breeding pools from which cultivars are developed by selfing and selection of desired phenotypes. Pedigree breeding is used commonly for the improvement of self-pollinating crops. Two parents that possess favorable, complementary traits are crossed to produce an Fi. An F2 population is produced by selfing one or several Fis. Selection of the best individuals may begin inAttorney Docket No.: B88552 1690 WO (00455)

[0212] the F2 population; then, beginning in the F3, the best individuals in the best families are selected. Replicated testing of families can begin in the F4 generation to improve the effectiveness of selection for traits with low heritability. At an advanced stage of inbreeding (i.e., Fe and F7), the best lines or mixtures of phenotypically similar lines are tested for potential release as new cultivars.

[0213] Using soybean cultivar 6073069 or 6167952 to develop other high energy density soybean varieties Soybean varieties such as high energy density soybean varieties including soybean cultivars 6073069 and 6167952 are typically developed for use in seed and grain production. However, soybean varieties such as soybean cultivar 6073069 or 6167952 also provide a source of breeding material that may be used to develop new soybean varieties. Plant breeding techniques known in the art and used in a soybean plant breeding program include, but are not limited to, recurrent selection, mass selection, bulk selection, mass selection, backcrossing, pedigree breeding, open pollination breeding, restriction fragment length polymorphism enhanced selection, genetic marker enhanced selection, making double haploids, and transformation. Often combinations of these techniques are used. The development of soybean varieties in a plant breeding program requires, in general, the development and evaluation of homozygous varieties. There are many analytical methods available to evaluate a new variety. The oldest and most traditional method of analysis is the observation of phenotypic traits, but genotypic analysis may also be used.

[0214] Additional Breeding Methods

[0215] One embodiment is directed to methods for producing a soybean plant by crossing a first parent soybean plant with a second parent soybean plant, wherein the first or second soybean plant is a high energy density soybean plant, including but not limited to the soybean plant from soybean cultivar 6073069 or 6167952. Further, both first and second parent soybean plants may be high energy density soybean plants, for example from soybean cultivar 6073069 or 6167952. Therefore, any methods using high energy density soybean plants, such as soybean cultivar 6073069 or 6167952 are part of the embodiments: selfing, backcrosses, hybrid breeding, and crosses to populations. Any plants produced using high energy density soybean plants such as soybean cultivar 6073069 or 6167952 as at least about one parent are also within the scope of the embodiments. Any such methods using a high energy density soybean plant, such as soybean variety 6073069 or 6167952 are part of the embodiments: selfing, sibbing, backcrosses, mass selection, pedigree breeding, bulk selection, hybrid production, crosses to populations, and the like. These methods are well known in the art and some of the more commonly used breeding methods are described herein. Descriptions of breeding methods can be found in one of several reference books (e.g., Allard, Principles of Plant Breeding (1960); Simmonds, Principles of CropAttorney Docket No.: B88552 1690 WO (00455)

[0216] Improvement (1979); Sneep, et al. (1979); Fehr, “Breeding Methods for Cultivar Development,” Chapter 7, Soybean Improvement, Production and Uses, 2nded., Wilcox editor (1987)).

[0217] The following describes breeding methods that may be used with high energy density soybean plants such as soybean cultivar 6073069 or 6167952 in the development of further soybean plants. One such embodiment is a method for developing a high energy density soybean progeny plant, such as a cultivar 6073069 or 6167952 progeny soybean plant in a soybean plant breeding program comprising: obtaining the soybean plant, or a part thereof, of a high energy density soybean plant, such as cultivar 6073069 or 6167952, utilizing said plant, or plant part, as a source of breeding material, and selecting a high energy density soybean progeny plant, such as a soybean cultivar 6073069 or 6167952 progeny plant with molecular markers in common with a high energy density soybean plant, such as cultivar 6073069 or 6167952 and / or with morphological and / or physiological characteristics of a high energy density soybean plant, such as cultivar 6073069 or 6167952. Breeding steps that may be used in the soybean plant breeding program include pedigree breeding, backcrossing, mutation breeding, and recurrent selection. In conjunction with these steps, techniques such as RFLP-enhanced selection, genetic marker enhanced selection (for example, SSR markers), and the making of double haploids may be utilized.

[0218] Another method involves producing a population of a high energy density soybean plants, such as soybean cultivar 6073069 or 6167952 progeny soybean plants, comprising crossing a high energy density soybean plant, such as cultivar 6073069 or 6167952 with another soybean plant, thereby producing a population of soybean plants which, on average, derive 50% of their alleles from the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952. A plant of this population may be selected and repeatedly selfed or sibbed with a soybean cultivar resulting from these successive filial generations. One embodiment is the soybean cultivar produced by this method and that has obtained at least about 50% of its alleles from the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952.

[0219] One of ordinary skill in the art of plant breeding would know how to evaluate the traits of two plant varieties to determine if there is no significant difference between the two traits expressed by those varieties. For example, see, Fehr and Walt, Principles of Cultivar Development, pp. 261-286 (1987). Thus, embodiments include high energy density soybean progeny plants, such as soybean cultivar 6073069 or 6167952 progeny soybean plants comprising a combination of at least about two cultivar 6073069 or 6167952 traits, so that said progeny soybean plant is not significantly different for said traits from the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 as determined at the 5% significance level when grown in the same environmental conditions. Using techniques described herein, molecular markers may be used to identify said progeny plant as a high energy density soybean progeny plant, such as a soybeanAttorney Docket No.: B88552 1690 WO (00455)

[0220] cultivar 6073069 or 6167952 progeny plant. Mean trait values may be used to determine whether trait differences are significant, and preferably the traits are measured on plants grown under the same environmental conditions. Once such a variety is developed, its value is substantial since it is important to advance the germplasm base as a whole in order to maintain or improve traits such as yield, disease resistance, pest resistance, and plant performance in extreme environmental conditions.

[0221] Progeny of a high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 may also be characterized through their filial relationship with the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952, as for example, being within a certain number of breeding crosses of the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952. A breeding cross is a cross made to introduce new genetics into the progeny, and is distinguished from a cross, such as a self or a sib cross, made to select among existing genetic alleles. The lower the number of breeding crosses in the pedigree, the closer the relationship between the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 and its progeny. For example, progeny produced by the methods described herein may be within 1, 2, 3, 4, or 5 breeding crosses of the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952.

[0222] Pedigree Breeding

[0223] Pedigree breeding starts with the crossing of two genotypes, such as soybean cultivar 6073069 or 6167952 and another soybean variety having one or more desirable characteristics that is lacking or which complements soybean cultivar 6073069 or 6167952. If the two original parents do not provide all the desired characteristics, other sources can be included in the breeding population. In the pedigree method, superior plants are selfed and selected in successive filial generations. In the succeeding filial generations, the heterozygous condition gives way to homogeneous varieties as a result of self-pollination and selection. Typically, in the pedigree method of breeding, five or more successive filial generations of selfing and selection is practiced: Fi to F2; F2 to F3; F3 to F4; F4 to F5; etc. After a sufficient amount of inbreeding, successive filial generations will serve to increase seed of the developed variety. Preferably, the developed variety comprises homozygous alleles at about 95% or more of its loci.

[0224] Backcross Breeding

[0225] Backcross breeding has been used to transfer genes for a simply inherited, highly heritable trait into a desirable homozygous cultivar or inbred line which is the recurrent parent. The source of the trait to be transferred is called the donor parent. After the initial cross, individuals possessing the phenotype of the donor parent are selected and repeatedly crossed (backcrossed) to the recurrentAttorney Docket No.: B88552 1690 WO (00455)

[0226] parent. The resulting plant is expected to have the attributes of the recurrent parent (e.g., cultivar) and the desirable trait transferred from the donor parent.

[0227] In addition to being used to create a backcross conversion, backcrossing can also be used in combination with pedigree breeding. As discussed previously, backcrossing can be used to transfer one or more specifically desirable traits from one variety, the donor parent, to a developed variety called the recurrent parent, which has overall good agronomic characteristics yet lacks that desirable trait or traits. However, the same procedure can be used to move the progeny toward the genotype of the recurrent parent, but at the same time retain many components of the nonrecurrent parent by stopping the backcrossing at an early stage and proceeding with selfing and selection. For example, a soybean variety may be crossed with another variety to produce a first-generation progeny plant. The first-generation progeny plant may then be backcrossed to one of its parent varieties to create a BCi or BC2. Progeny are selfed and selected so that the newly developed variety has many of the attributes of the recurrent parent and yet several of the desired attributes of the nonrecurrent parent. This approach leverages the value and strengths of the recurrent parent for use in new soybean varieties.

[0228] Therefore, an embodiment is a method of making a backcross conversion of a high energy density soybean plant, such as soybean variety 6073069 or 6167952, comprising the steps of crossing a high energy density soybean plant, such as soybean variety 6073069 or 6167952 with a donor plant comprising a desired trait, selecting an Fi progeny plant comprising the desired trait, and backcrossing the selected F 1 progeny plant to a high energy density soybean plant, such as soybean variety 6073069 or 6167952. This method may further comprise the step of obtaining a molecular marker profile of a high energy density soybean plant, such as soybean variety 6073069 or 6167952 and using the molecular marker profile to select for a progeny plant with the desired trait and the molecular marker profile of a high energy density soybean plant, such as soybean cultivar 6073069 or 6167952. In one embodiment, the desired trait is a mutant gene, gene, or transgene present in the donor parent.

[0229] Recurrent Selection and Mass Selection

[0230] Recurrent selection is a method used in a plant breeding program to improve a population of plants. A high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 is suitable for use in a recurrent selection program. The method entails individual plants cross pollinating with each other to form progeny. The progeny are grown and the superior progeny selected by any number of selection methods, which include individual plant, half-sib progeny, full-sib progeny, and selfed progeny. The selected progeny are cross pollinated with each other to form progeny for another population. This population is planted and again superior plants are selected toAttorney Docket No.: B88552 1690 WO (00455)

[0231] cross pollinate with each other. Recurrent selection is a cyclical process and therefore can be repeated as many times as desired. The objective of recurrent selection is to improve the traits of a population. The improved population can then be used as a source of breeding material to obtain new varieties for commercial or breeding use, including the production of a synthetic cultivar. A synthetic cultivar is the resultant progeny formed by the intercrossing of several selected varieties.

[0232] Mass selection is a useful technique when used in conjunction with molecular marker enhanced selection. In mass selection, seeds from individuals are selected based on phenotype or genotype. These selected seeds are then bulked and used to grow the next generation. Bulk selection requires growing a population of plants in a bulk plot, allowing the plants to selfpollinate, harvesting the seed in bulk, and then using a sample of the seed harvested in bulk to plant the next generation. Also, instead of self-pollination, directed pollination could be used as part of the breeding program.

[0233] Mass and recurrent selections can be used to improve populations of either self- or crosspollinating crops. A genetically variable population of heterozygous individuals is either identified, or created, by intercrossing several different parents. The plants are selected based on individual superiority, outstanding progeny, or excellent combining ability. The selected plants are intercrossed to produce a new population in which further cycles of selection are continued.

[0234] Single-Seed Descent

[0235] The single-seed descent procedure in the strict sense refers to planting a segregating population, harvesting a sample of one seed per plant, and using the one-seed sample to plant the next generation. When the population has been advanced from the F2 to the desired level of inbreeding, the plants from which lines are derived will each trace to different F2 individuals. The number of plants in a population declines each generation due to failure of some seeds to germinate or some plants to produce at least about one seed. As a result, not all of the F2 plants originally sampled in the population will be represented by a progeny when generation advance is completed.

[0236] Multiple-Seed Procedure

[0237] In a multiple-seed procedure, soybean breeders commonly harvest one or more pods from each plant in a population and thresh them together to form a bulk. Part of the bulk is used to plant the next generation and part is put in reserve. The procedure has been referred to as modified single-seed descent or the pod-bulk technique.

[0238] The multiple-seed procedure has been used to save labor at harvest. It is considerably faster to thresh pods with a machine than to remove one seed from each by hand for the single-seed procedure. The multiple-seed procedure also makes it possible to plant the same number of seeds of a population in each generation of inbreeding. Enough seeds are harvested to make up for thoseAttorney Docket No.: B88552 1690 WO (00455)

[0239] plants that did not germinate or produce seed.

[0240] Breeding with Molecular Markers

[0241] Molecular markers, which includes markers identified through the use of techniques such as Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs), and Single Nucleotide Polymorphisms (SNPs), may be used in plant breeding methods utilizing high energy density soybean plants, such as soybean cultivar 6073069 or 6167952.

[0242] Isozyme Electrophoresis and RFLPs have been widely used to determine genetic composition. Shoemaker and Olsen, Molecular Linkage Map of Soybean (Glycine max L. Merr.), pp. 6.131-6.138 (1993). In S. J. O’Brien (ed.), Genetic Maps: Locus Maps of Complex Genomes, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., developed a molecular genetic linkage map that consisted of 25 linkage groups with about 365 RFLP, 11 RAPD (random amplified polymorphic DNA), 3 classical markers, and 4 isozyme loci. See also, Shoemaker, R. C., 1994 RFLP Map of Soybean, pp. 299-309; In R. L. Phillips and I. K. Vasil (ed.), DNA-based markers in plants, Kluwer Academic Press Dordrecht, the Netherlands.

[0243] SSR technology is currently the most efficient and practical marker technology. More marker loci can be routinely used, and more alleles per marker locus can be found, using SSRs in comparison to RFLPs. For example, Diwan and Cregan described highly polymorphic microsatellite loci in soybean with as many as 26 alleles. (Diwan, N., and Cregan. P. B., Automated sizing of fluorescent-labeled simple sequence repeat (SSR) markers to assay genetic variation in Soybean, Theor. Appl. Genet., 95:220-225 (1997)). Single Nucleotide Polymorphisms may also be used to identify the unique genetic composition of the embodiment(s) and progeny varieties retaining that unique genetic composition. Various molecular marker techniques may be used in combination to enhance overall resolution.

[0244] Soybean DNA molecular marker linkage maps have been rapidly constructed and widely implemented in genetic studies. One such study is described in Cregan, et. al, “An Integrated Genetic Linkage Map of the Soybean Genome,” Crop Science, 39:1464-1490 (1999). Sequences and PCR conditions of SSR Loci in Soybean, as well as the most current genetic map, may be found in Soybase on the World Wide Web.

[0245] Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed. One use of molecular markers is Quantitative Trait Loci (QTL) mapping. QTL mapping is the use of markers, which are known toAttorney Docket No.: B88552 1690 WO (00455)

[0246] be closely linked to alleles that have measurable effects on a quantitative trait. Selection in the breeding process is based upon the accumulation of markers linked to the positive effecting alleles and / or the elimination of the markers linked to the negative effecting alleles from the plant’s genome.

[0247] Molecular markers can also be used during the breeding process for the selection of qualitative traits. For example, markers closely linked to alleles or markers containing sequences within the actual alleles of interest can be used to select plants that contain the alleles of interest during a backcrossing breeding program. The markers can also be used to select for the genome of the recurrent parent and against the genome of the donor parent. Using this procedure can minimize the amount of genome from the donor parent that remains in the selected plants. It can also be used to reduce the number of crosses back to the recurrent parent needed in a backcrossing program. The use of molecular markers in the selection process is often called genetic marker enhanced selection. Molecular markers may also be used to identify and exclude certain sources of germplasm as parental varieties or ancestors of a plant by providing a means of tracking genetic profiles through crosses.

[0248] Production of Double Haploids

[0249] The production of double haploids can also be used for the development of plants with a homozygous phenotype in the breeding program. For example, a soybean plant for which a high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 is a parent can be used to produce double haploid plants. Double haploids are produced by the doubling of a set of chromosomes (IN) from a heterozygous plant to produce a completely homozygous individual. For example, see, Wan, et al., “Efficient Production of Doubled Haploid Plants Through Colchicine Treatment of Anther-Derived Maize Callus,” Theoretical and Applied Genetics, 77:889-892 (1989) and U.S. Pat. No. 7,135,615. This can be advantageous because the process omits the generations of selfing needed to obtain a homozygous plant from a heterozygous source.

[0250] Haploid induction systems have been developed for various plants to produce haploid tissues, plants and seeds. The haploid induction system can produce haploid plants from any genotype by crossing a selected line (as female) with an inducer line. Such inducer lines for maize include Stock 6 (Coe, Am. Nat., 93:381-382 (1959); Sharkar and Coe, Genetics, 54:453-464 (1966); KEMS (Deimling, Roeber, and Geiger, Vortr. Pflanzenzuchtg, 38:203-224 (1997); or KMS and ZMS (Chalyk, Bylich & Chebotar, MNL, 68:47 (1994); Chalyk & Chebotar, Plant Breeding, 119:363-364 (2000)); and indeterminate gametophyte (ig) mutation (Kermicle, Science, 166:1422- 1424 (1969)). The disclosures of which are incorporated herein by reference.

[0251] Methods for obtaining haploid plants are also disclosed in Kobayashi, M., et al., Joum. ofAttorney Docket No.: B88552 1690 WO (00455)

[0252] Heredity, 71(1):9-14 (1980); Pollacsek, M., Agronomic (Paris) 12(3):247-251 (1992); Cho-Un-Haing, etal., Joum. of Plant Biol., 39(3): 185-188 (1996); Verdoodt, L„ et al., 96(2):294-300 (February 1998); Chalyk, et al., Maize Genet Coop., Newsletter 68:47 (1994).

[0253] Thus, an embodiment is a process for making a substantially homozygous high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 progeny plant by producing or obtaining a seed from the cross of a high energy density soybean plant, such as soybean cultivar 6073069 or 6167952 and another soybean plant and applying double haploid methods to the Fl seed or Fl plant or to any successive filial generation. Based on studies in maize and currently being conducted in soybean, such methods would decrease the number of generations required to produce a variety with similar genetics or characteristics to the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952. See, Bernardo, R. and Kahler, A. L., Theor. Appl. Genet., 102:986-992 (2001).

[0254] In particular, a process of making seed retaining the molecular marker profile of a high energy density soybean plant, such as soybean variety 6073069 or 6167952 is contemplated, such process comprising obtaining or producing F 1 seed for which the high energy density soybean plant, such as soybean variety 6073069 or 6167952 is a parent, inducing doubled haploids to create progeny without the occurrence of meiotic segregation, obtaining the molecular marker profile of the high energy density soybean plant, such as soybean variety 6073069 or 6167952, and selecting progeny that retain the molecular marker profile of the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952.

[0255] Descriptions of other breeding methods that are commonly used for different traits and crops can be found in one of several reference books (e.g., Allard (1960); Simmonds (1979);

[0256] Sneep, et al. (1979); Fehr (1987)).

[0257] Tissue Culture

[0258] Further reproduction of the variety can occur by tissue culture and regeneration. Tissue culture of various tissues of soybeans and regeneration of plants therefrom is well-known and widely published. For example, reference may be had to Komatsuda, T., et al., Crop Set., 31 : 333 -337 (1991); Stephens, P. A., et al., Theor. Appl. Genet., 82:633-635 (1991); Komatsuda, T., et al., Plant Cell, Tissue and Organ Culture, 28: 103-113 (1992); Dhir, S., et al., Plant Cell Reports, 11:285-289 (1992); Pandey, P., et al., Japan J. Breed., 42:1-5 (1992); and Shetty, K., et al., Plant Science, 81:245-251 (1992); as well as U.S. Pat. Nos. 5,024,944 and U.S. Pat. No. 5,008,200. Thus, another aspect or embodiment is to provide cells which upon growth and differentiation produce soybean plants having the physiological and morphological characteristics of the high energy density soybean plant, such as soybean cultivar 6073069 or 6167952.Attorney Docket No.: B88552 1690 WO (00455)

[0259] Regeneration refers to the development of a plant from tissue culture. The term “tissue culture” indicates a composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant. Exemplary types of tissue cultures are protoplasts, calli, plant clumps, and plant cells that can generate tissue culture that are intact in plants or parts of plants, such as embryos, pollen, flowers, seeds, pods, petioles, leaves, stems, roots, root tips, anthers, pistils, and the like. Means for preparing and maintaining plant tissue culture are well known in the art. By way of example, a tissue culture comprising organs has been used to produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and 5,977,445 describe certain techniques, the disclosures of which are incorporated herein by reference.

[0260] V. Producing Soybean Meal Composition

[0261] A method of producing a soybean meal composition. The method includes processing a population of soybean seeds to provide the soybean meal composition. The population of soybean seeds contains protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis. The soybean meal composition produced from the population of soybean seeds contains true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM) or TMEn that is greater at least by 10% relative to a control soybean meal composition, e.g., derived from control, reference, or commodity soybean seeds.

[0262] In embodiments of the methods provided herein, high energy density soybean seeds with seed protein content of at least about 39% (dry basis) and seed sucrose content of at least about 8.5% (dry bases) are cleaned to remove foreign matter and loose hulls. The cleaned soybeans are processed using suitable methods to produce a soybean meal composition of choice. For example, soybeans are processed by fermenting, hexane extraction, drying, dehulling, grinding, heating, toasting, flaking, cracking, and / or defatting to produce a soybean meal composition.

[0263] In one example, the soybeans are fermented and an energy-enriched soybean meal composition is isolated (e.g., by hexane extraction) from the fermented soybeans.

[0264] In another example, the soybeans are ground to provide a ground soybean composition, and an energy-enriched soybean meal composition is isolated (e.g., by hexane extraction) from the ground soybean composition.

[0265] In further example, the whole soybeans are heated to remove excess moisture. The dried beans are cracked and dehulled to remove the skin covering the soybeans and produce soy grits. The resulting soy grits are ground in hot water (80°C) to produce a soy slurry. Following the grinding process, the soy slurry is boiled and stirred to deactivate undesirable enzymes and remove volatile, unsavory flavor compounds. Once the slurry isAttorney Docket No.: B88552 1690 WO (00455)

[0266] cooled to room temperature, it is filtered to separate undispersed fiber residue. The liquid is collected from the filter to afford soybean meal.

[0267] The general processes described herein for producing soybean meal composition are varied in each embodiment to optimize the desired characteristics of each soybean meal composition. Different soybean varieties and / or ratios of soybeans to processing agents are used to optimize characteristics such as soybean meal energy, protein, and sucrose levels, flavor profiles, and thickness. Additional considerations include other sensory attributes, soymilk yield, soybean inclusion, and other nutritional values such as lipids and fatty acids, other sugars and oligosaccharides, fibers, and minerals.

[0268] In the methods provided herein, a population of high energy density soybeans provided herein can be processed to produce a high energy density soybean meal composition. Content of a composition (e.g., protein content, sucrose content, oil content, oligosaccharide content, e.g., X% dry basis) in a population of soybean seeds as used herein may refer to average content (e.g., average protein content) of X% dry basis, at least a portion of the soybean seeds having the content (e.g., protein content) of X% dry basis, or the majority of the soybean seeds having the content (e.g., protein content) of X% dry basis. For example, the population of high energy density soybean seeds for use in the methods provided herein can contain protein content of protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 9% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 9.5% dry basis; protein content of at least about 39% dry basis and sucrose content of at least about 10% dry basis; protein content of at least about 40% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 41% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 42% dry basis and sucrose content of at least about 8.5% dry basis; protein content of at least about 43% dry basis and sucrose content of at least about 8.5% dry basis; or protein content of at least about 44% dry basis and sucrose content of at least about 8.5% dry basis.

[0269] The population of soybean seeds for use in the methods provided herein can contain oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.5% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 9% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 9.5% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 10% dry basis; oil content of at least about 22% dry basis, proteinAttorney Docket No.: B88552 1690 WO (00455)

[0270] content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 23% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 24% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 25% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis; or oil content of at least about 26% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 42% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 43% dry basis, and sucrose content of at least about 8.25% dry basis; oil content of at least about 21.5% dry basis, protein content of at least about 44% dry basis, and sucrose content of at least about 8.25% dry basis.

[0271] In some embodiments, the population of soybean seeds for use in the methods provided herein comprises oligosaccharide content of about 1% or less, such as about 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, 0.1% or less, about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, about 1-0.7%, 0.7%-0.4%, or 0.4-0.1%.

[0272] In some embodiments, the population of soybean seeds for use in the methods provided herein contains (i) protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis or (ii) oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis. In some embodiments, the population of soybean seeds contains seeds of soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008; or an offspring plant or a parent plant of the soybean cultivar 6073069 or the soybean cultivar 6167952.

[0273] In some embodiments, the soybean meal composition produced according the methods provided herein contains TMEn of at least about 1350 kcal / kg DM or TMEn that is greater by at least 10% relative to a control soybean meal composition derived from, e.g., reference or commodity soybean seeds, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

[0274] The methods provided herein can process a population of high energy density soybeans and produce high energy density soybean meal composition that has higher energy density (e.g., high protein content and / or high sucrose content) relative to the population of soybean seeds from which the soybean meal composition was derived. For example, the high energy density soybean mealAttorney Docket No.: B88552 1690 WO (00455)

[0275] composition can have at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more TMEn per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained. Additionally or alternatively, the high energy density soybean meal composition can have at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more protein content per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained. The high energy density soybean meal composition can have at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more sucrose content per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained. Additionally or alternatively, the high energy density soybean meal composition can have at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, or at least about 1.6 times more TMEn per unit weight relative to the soybeans from which the high energy density soybean meal composition was obtained.

[0276] Also provided is a soybean meal composition produced by the methods provided herein. The soybean meal composition produced by the methods provided herein can be high density soybean meal composition, having at least about one characteristics described herein. For example, the soybean meal composition produced by the methods provided herein can contain TMEn of at least about at least about 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500 kcal / kg DM, about 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500 kcal / kg DM or more, or about 1300-1350, 1350-1400, 1400-1450, or 1450-1500 kcal / kg DM; TMEn that is greater by at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more, by about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or more, or by about 5-10%, 7-13%, or 10-15% relative to a control or reference soybean meal composition; protein content of at least about 62% dry basis; oligosaccharide content of about 1% or less dry basis; sucrose content of at least about 10.5% dry basis; ; and / or protein content of at least about 57.5% dry basis and sucrose content of at least about 11.5% dry basis. Other characteristics of a soybean meal composition produced from the population of soybean seeds are described elsewhere herein.

[0277] In some embodiments, the soybean meal composition produced by the methods provided herein comprises feed, protein isolate, protein concentrate, soybean flour, protein flakes, and / or textured soy protein (TSP).Attorney Docket No.: B88552 1690 WO (00455)

[0278] EXAMPLES

[0279] The presently disclosed subject matter will be further understood by reference to the following Examples, which are provided as example of the invention, and not by way of limitation.

[0280] Example 1: Preparation of soybean meal compositions from high energy soybean varieties This example describes the production of soybean meal compositions from different source high energy density soybean varieties having high seed protein content and high seed sucrose content as compared to commodity soybeans.

[0281] High energy density soybean seeds with seed protein content of at least about 39% (dry basis) and seed sucrose content of at least about 8.5% (dry bases) were cleaned to remove foreign matter and loose hulls. An energy-enriched soybean meal composition was isolated (e.g., by hexane extraction) from the cleaned soybeans.

[0282] The seed protein and sucrose content as weight percentages of the soybean (% w / w, on a dry basis) for the high energy density soybean varieties and a commodity line, as well as protein and sucrose content and nitrogen-corrected true metabolizable energy (TMEn) of the soybean meal compositions prepared from each soybean variety is shown in Table 1. TMEn was calculated based on the correlation between the protein and sucrose content (% dry basis) and TMEn (kcal / kg dry matter) available in Parsons & Araba 2000 Poultry Science 79(8): 1127-31 and Baker et al, 2011 Poultry Science 90(2):390-5, the entire contents of which references are incorporated herein by reference. In these studies, TMEn was measured using standard methodology in a precision-fed rooster assay. In brief, in the precision-fed assay, roosters are usually fasted for 24-26 h and then crop-intubated with approximately 30 g of a test ingredient, followed by being fasted for an additional 48 h during which time excreta are collected quantitatively.

[0283] As shown in Table 1, the soybean meal produced from the high energy density soybean seeds provided herein has higher protein content, higher sucrose content, and higher TMEn as compared to the soybean meal produced from the commodity soybean seeds.

[0284] Soybean meal compositions as generally described in Example 2 can offer a plant-based meal with higher energy density than commercially available products.

[0285] Table 1. Protein and sucrose content in high energy soybean varieties and soybean meal compositions prepared from the varieties

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[0287]

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[0290]

[0291] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described in any way.

[0292] It is appreciated that certain features of the disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination or as suitable in any other described embodiment of the disclosure. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

[0293] While various aspects of the invention are described herein, it is not intended that the invention be limited by any particular aspect. On the contrary, the invention encompasses various alternatives, modifications, and equivalents, as will be appreciated by those of skill in the art. Furthermore, where feasible, any of the aspects disclosed herein may be combined with each other (e.g., the feature according to one aspect may be added to the features of another aspect or replace an equivalent feature of another aspect) or with features that are well known in the art, unless indicated otherwise by context.

Claims

Attorney Docket No.: B88552 1690 WO (00455)What is claimed is:

1. A soybean meal composition comprising nitrogen-corrected true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM).

2. The soybean meal composition of claim 1, comprising sucrose content of at least about 10.5% dry basis.

3. The soybean meal composition of claim 1, comprising the TMEn that is greater by at least about 10% relative to a control soybean meal composition derived from a commodity soybean cultivar.

4. The soybean meal composition of claim 1, comprising TMEn of at least about 1350 kcal / kg DM.

5. The soybean meal composition of claim 4, comprising protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

6. The soybean meal composition of any one of claims 1-5, comprising protein content of at least about 62% dry basis.

7. The soybean meal composition of any one of claims 1-6, comprising oligosaccharide content of about 1% or less dry basis.

8. The soybean meal of any one of claims 1-7, wherein said soybean meal comprises soybean meal derived from:soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007; orsoybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008.

9. The soybean meal composition of any one of claims 1-8, comprising a de-fatted soybean meal composition.

10. A population of soybean seeds, comprising protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis.Attorney Docket No.: B88552 1690 WO (00455)11. A population of soybean seeds, comprising oil content of 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis.

12. The population of soybean seeds of claim 10 or 11, comprising protein content of at least about 44% dry basis.

13. The population of soybean seeds of any one of claims 10-12, comprising oligosaccharide content of about 1% or less.

14. The population of soybean seeds of any one of claims 10-13, wherein said population of soybean seeds comprises seeds of:soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007,soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008, oran offspring plant or a parent plant of the soybean cultivar 6073069 or the soybean cultivar 6167952.

15. A soybean meal composition produced from the population of soybean seeds of any one of claims 10-14.

16. The soybean meal composition of claim 15, comprising nitrogen-corrected true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM) or TMEn that is greater by at least about 10% relative to a control soybean meal composition derived from a commodity soybean cultivar.

17. The soybean meal composition of claim 15 or 16, comprisingprotein content of at least about 62% dry basis;oligosaccharide content of about 1% or less dry basis;sucrose content of at least about 10.5% dry basis;and / orTMEn of at least about 1350 kcal / kg, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.Attorney Docket No.: B88552 1690 WO (00455)18. The soybean meal composition of any one of claims 15-17, comprising a de-fatted soybean meal.

19. A method for producing a soybean meal composition, the method comprising processing a population of soybean seeds to provide the soybean meal composition, wherein the population of soybean seeds comprises (i) protein content of at least about 39% dry basis and sucrose content of at least about 8.5% dry basis or (ii) oil content of at least about 21.5% dry basis, protein content of at least about 41% dry basis, and sucrose content of at least about 8.25% dry basis, and wherein the soybean meal composition comprises nitrogen-corrected true metabolizable energy (TMEn) of at least about 1300 kcal / kg dry matter (kg DM) or TMEn that is greater by at least about 10% relative to a control soybean meal composition derived from a commodity soybean cultivar.

20. The method of claim 19, wherein the population of soybean seeds comprises protein content of at least about 44% dry basis.

21. The method of claim 19 or 20, wherein the population of soybean seeds comprises oligosaccharide content of about 1% or less.

22. The method of any one of claims 19-21, wherein the population of soybean seeds comprises seeds of:soybean cultivar 6073069, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412007,soybean cultivar 6167952, wherein a representative sample of seed of said cultivar is deposited under NCMA Accession Number 202412008, oran offspring plant or a parent plant of the soybean cultivar 6073069 or the soybean cultivar 6167952.

23. The method of any one of claims 19-22, wherein the soybean meal composition comprises TMEn of at least about 1350 kcal / kg DM, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

24. The method of any one of claims 19-23, wherein processing the population of soybean seeds comprises fermenting, hexane extraction, drying, dehulling, grinding, heating, toasting, flaking, cracking, and / or defatting of the population of soybean seeds.Attorney Docket No.: B88552 1690 WO (00455)25. A soybean meal composition produced by the method of any one of claims 19-24.

26. The soybean meal composition of claim 25, comprisingprotein content of at least about 62% dry basis;oligosaccharide content of about 1% or less dry basis;sucrose content of at least about 10.5% dry basis;and / orTMEn of at least about 1350 kcal / kg, protein content of at least about 57.5% dry basis, and sucrose content of at least about 11.5% dry basis.

27. The soybean meal composition of claim 25 or 26, comprising a de-fatted soybean meal composition.

28. The soybean meal composition of any one of claims 1-9, 15-18, and 25-27, wherein the soybean meal composition comprises feed, protein isolate, protein concentrate, soybean flour, protein flakes, and / or textured soy protein (TSP).

29. A feed, food, or beverage product produced from the soybean composition of any one of claims 1-9, 15-18, and 25-28 or the population of soybean seeds of any one of claims 10-14.

30. The feed, food, or beverage product of claim 29, wherein the feed, food, or beverage product comprises animal feed, aquatic animal feed, soy alternative meat products, soy alternative egg products, soy alternative dairy products, soy energy bars, soy food products, soy infant formula, or soy baby foods.

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