Renewable agricultural compositions

Abstract

Disclosed are agricultural compositions derived from a renewable source and methods of making said agricultural compositions. The agricultural compositions may comprise an extract derived from non-woody plants of the genus Hesperaloe. The extract of a non-woody plant of the genus Hesperaloe may comprise water soluble components including lipids, proteins, inorganic salts, saccharides, polysaccharides, organic acids, saponins, or combinations thereof. The extracts may be prepared with minimal processing to remove at least a portion of the water insoluble solids. The agricultural compositions have a high degree of efficacy against a range of agricultural pests and may be particularly useful in preventing or inhibiting the growth of fungal organisms.

Description

RENEWABLE AGRICULTURAL COMPOSITIONSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 532,083, filed August 11, 2023, which is incorporated herein in its entirety.FIELD

[0002] This application relates to extracts from non-woody plants of the genus Hesperaloe such as, but not limited to, extracts comprising saponin.BACKGROUND

[0003] Agricultural compositions comprising agricultural chemical and / or biological agent active ingredients, including those found in fertilizers, insecticides, fungicides, biofungicides, herbicides, miticides, and plant grow th regulators, can be incorporated into a finished formulation before being applied to soil or growth medium, plants, plant parts, and / or seeds. Diluents and / or additives can be chosen according to the chemical properties and end-use applications of the agricultural compositions. For example, various materials are used in formulations to optimally dilute, formulate, and / or solubilize such active ingredients and improve the properties of the finished formulation, such as for example: storage stability', ease of handling, enhanced penetration into the soil, plant, pest (e.g., insect), or seed, an improved safety profile of the active ingredient, or improved active ingredient efficacy against target organisms.

[0004] Traditionally many of the chemicals used in agricultural compositions have been geologically derived and often involve complex and costly synthesis. Moreover, intensive and / or inappropriate use of chemicals causes ecosystem contamination and adverse health effects; pesticide residues in food endanger human health, along with their accumulation in the environment, affect non-target organisms in a negative way, and endanger the future of plant production by the development of pest resistance. Increasingly agricultural chemicals are being replaced with microbiological, biochemical, and macrobiological plant protection products for the control of pests, weeds, and disease-causing agents.

[0005] Biochemical pesticides, which are derived in whole or in part from renewable biomass resources, are of particular interest. Biomass resources are organic materials that are available on a renewable or recurring basis such as crop residues, wood residues, grasses, and aquatic plants. One example of biochemical pesticides are essential oils, particularly oils derived from aromatic, medicinal and / or seasoning plants through different distillation and / or solvent extraction processes. Essential oils often comprise complex mixtures of mono terpenes, sesquiterpenes and aromatics.

[0006] Currently, a major limitation to the broad exploitation of biochemical pesticides is the fact that commercially availability remains scarce and they are expensive, particularly compared to geologically derived chemicals. The expense is due in large part to the limited number of plant extracts having significant amounts of active agents and the scarcity of widespread cultivation of such plants in significant enough amounts to reduce cost.

[0007] Thus, there is a need for the development of biochemical pesticides, and in particular, an agricultural composition that includes a significant proportion of its ingredients from biomass resources rather than geologically derived chemicals.SUMMARY

[0008] Described herein are agricultural compositions that include a significant proportion of ingredients from biomass resources and can be produced on a scale and at a cost comparable to geologically derived chemicals. The agricultural compositions and methods of using the same, may comprise an extract derived from a non-woody plant of the genus Hesperaloe. The agricultural compositions, which in certain embodiments comprise a pesticidal composition, may be formulated as a liquid spray, crystals, gel, and / or pellet. The agricultural compositions may comprise an impregnating material, such as posts. The agricultural compositions may be applied to a broad range of crops to inhibit or protect against a wide range of pests. Thus, in certain embodiments, the present disclosure is directed to a method for controlling pests by applying a pesticidally-effective amount of the pesticidal compositions to a location where pest control is desired.

[0009] In some cases, the agricultural composition may comprise water soluble solids extracted from Hesperaloe, where the extract may comprise one or more of a lipid, a protein,an inorganic salt, a saccharide, a polysaccharide, a saponin, and / or an organic acid. In certain examples, the agricultural composition may be provided as a liquid composition that may be topically applied to a plant by spraying to improve resistance to a pest or organism, particularly a fungi or bacteria.

[0010] In some examples, an agricultural composition may comprise a Hesperaloe extract comprising one or more phenolic compounds (such as tannins, quercetin, leucocyanidin, kaempferol, among others), organic acids (such as caffeic acid, gallic acid, coumaric acid), free saccharides, polysaccharides, lipids, and nitrogen-containing compounds, such as proteins.

[0011] Also disclosed herein are methods of preparing an agricultural composition from Hesperaloe biomass. The method may comprise milling biomass derived from non-woody plants of the genus Hesperaloe including, for example, Hesperaloe funifera, Hesperaloe nocturna, Hesperaloe parviflora, and Hesperaloe chiangii. The method may further comprise extracting the biomass with a first solvent to yield a crude extract, mixing the crude extract with a second solvent to form a first precipitate, and then removing the first precipitate to yield a clarified extract. The clarified extract may comprise water soluble solids including one or more lipids, proteins, inorganic salts, saccharides, polysaccharides, organic acids or saponins.

[0012] In some examples, the water soluble fraction extracted from Hesperaloe biomass may treated to remove at least a portion of the dissolved solids and yield a clarified extract comprising water soluble solids including one or more lipids, proteins, inorganic salts, saccharides, polysaccharides, organic acids or saponins. The treatment step may comprise the steps of mixing the crude Hesperaloe extract with a solvent to form a first solution, adjusting the pH of the first solution, e.g., from about 6 to about 7, forming a first precipitate, and removing the first precipitate, such as by filtration, to yield a clarified extract.DESCRIPTION OF THE DRAWINGS

[0013] The disclosure may be better understood by reference to the following nonlimiting drawings.

[0014] FIGS. 1 A and IB illustrate a triterpenoid saponin and a steroidal saponin, respectively.

[0015] FIGS. 2A-C illustrate various novel saponins extracted from non-woody plants of the genus Hesperaloe according to the present invention including. 25(27)- dehydrofucreastatin (FIG. 2A); 5(6),25(27)-disdehydroyuccaloiside C (FIG. 2B); and 5(6)- disdehydroyuccaloiside C (FIG. 2C).

[0016] FIG. 3 provides a schematic flow diagram for example multistage processes according to some embodiments of the present disclosure.

[0017] FIG. 4 provides a schematic flow diagram for an example method according to some embodiments of the present disclosure.DEFINITIONS

[0018] As used herein, the articles “a,” “an,” and "the" can be used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article, unless the language and / or context clearly indicates otherw ise. By way of example, “an element” means one element or more than one element. Furthermore, as used herein, the term “about” as used in connection with a numerical value throughout the specification and the claims denotes an interval of accuracy, familiar and acceptable to a person skilled in the art. In general, such interval of accuracy is ±10%. All numbers in this description indicating amounts, ratios of materials, physical properties of materials, and / or use are to be understood as modified by the word “about.” except as otherwise explicitly indicated. As an additional matter, all percentage amounts of the components of the cleaning composition are by weight percent of the overall composition, unless otherwise specified.

[0019] As used herein, the terms “include”, “includes” and “including” are meant to be non-limiting.

[0020] The term “renewable” is synonymous with the terms “biobased.” “sustainable,” “sustainably derived,” or “from sustainable sources” and means bio-derived (derived from a renewable resource, e.g., plants) or “non-geologically derived.”

[0021] “Geologically derived” means derived from, for example, petrochemicals, natural gas, or coal. “Geologically derived” materials cannot be easily replenished or regrown (e.g., in contrast to plant- or algae-produced oils).

[0022] As used herein, the term “renewable component” refers to a component that is derived from renewable feedstock and contains renewable carbon. A renewable feedstock is a feedstock that is derived from a renewable resource, e.g., plants, and non-geologically derived. A material may be partially renewable (less than 100% renewable carbon content, from about 1% to about 30%, 40%, 50%, 60%, 70%, 80%, or 90% renewable carbon content) or 100% renewable (100% renewable carbon content). A renewable material may be blended with a nonrenewable material.

[0023] The term “renewable carbon” generally refers to the bio-derived carbon determined in accordance with ASTM D6866, which distinguishes carbon resulting from renewable inputs from those derived from geologically derived inputs.

[0024] The term “bio-based carbon content” generally refers to a measure of the amount of carbon present in a product that is derived from a renewable source, such as plant, animal, marine, or forestry feedstocks.

[0025] As used herein, the term “biomass” generally refers to whole plants and plant organs (i.e., leaves, stems, flowers, roots, etc.) of the genus Hesperaloe including, for example, Hesperaloe fiinifera, Hesperaloe nocturna, Hesperaloe parviflora, and Hesperaloe chiangii. In some instances, water-soluble solids may be prepared from biomass comprising an above ground portion of the Hesperaloe plant, and more particularly the portion of the Hesperaloe plant above the crown, and still more preferable the leaves of the Hesperaloe plant.

[0026] As used herein, the term “saccharide” is used interchangeably with the terms “polysaccharide,” “oligosaccharide,” and “sugar,” the definitions of which are well known to those skilled in the art of carbohydrate chemistry. It should be noted that saccharides can be in the form of mono-, oligo- and / or polysaccharides. Preferably saccharides are water-soluble and do not include cellulose, hemicellulose or mono-, oligo- and / or polysaccharides bound to other compounds, such as glycosides (arabinose, glucose, galactose, xylose, and glucuronic acid) bound to a triterpenoid to form a saponin.

[0027] As used herein the term "saponin" generally refers to a glycoside comprising a sugar component referred to as a glycone and a non-sugar component referred to as an aglycone. Depending on the structure of the aglycone, the saponin may be classified as a triterpenoid saponin, illustrated in FIG. 1A. or as a steroidal saponin, illustrated in FIG. IB. The aglycone portion of the saponin may be either a pentacyclic triterpenoid or a tetracyclic triterpenoid, both of which contain 30 carbon atoms. Whether steroidal or triterpenoid, saponins may be mono, bi or tridesmodic. Monodesmodic saponins have a single saccharide, normally attached at C-3. Bidesmodic saponins have two saccharides, often with one attached through an ether linkage at C-3 and the other either attached through an ester linkage at C-28 or through an ether linkage at C-20 (pentacyclic and tetracyclic triterpene saponins, respectively), or through an ether linkage at C-26 (furostane saponins). In certain instances, Hesperaloe biomass may comprise at least about 5 wt. % of total saponins, such as from about 5 to about 25 wt. %, or about 5 to about 15 wt. %, or from about 8 to about 12 wt. %, based upon the bone-dry weight of the biomass.

[0028] As used herein the term “surfactant” generally refers to a substance or compound that reduces surface tension when dissolved in water or water solutions, or that reduces interfacial tension between two liquids, or between a liquid and a solid and includes cationic, anionic, nonionic, zwitterionic, amphoteric agents and / or combinations thereof. Surfactants are composed of a polar part and a non-polar part.

[0029] As used herein, an anionic surfactant are surfactants that carry a negatively charged functional (i.e. , head) group.

[0030] As used herein, cationic surfactants are surfactants that have a positively charged functional group.

[0031] As used herein, non-ionic surfactants are surfactants that have a polar head group that is not electrically charged. Mon-ionic surfactants usually rely on a functional group able to deprotonate but only to a very low degree.

[0032] As used herein, amphoteric surfactants are surfactants with a dual charge. The ionic charge of an amphoteric surfactant can change between anionic properties, an isoeletric neutral stage and cationic properties depending upon the pH value. Amphoteric surfactants have characteristics of stability against electrolytes, acids, alkalis, and hard water.

[0033] As used herein, zwiterionic surfactants are surfactants with an equal number of positively and negatively charged functional groups resulting in an overall neutral charge. Zwiterionic surfactants have characteristics of high-water solubility, high surface activity, and thermal and chemical stability7.

[0034] As used herein the term "water soluble solids" generally refers to dry mater which remains after the extract has been centrifuged, filtered and all water is evaporated. The procedure for measuring water soluble solids of a biomass extract of the present invention is described in detail in the Examples provided herein. Water soluble solids may be expressed on a percentage basis relative to the mass of bone dry biomass. Dry weight means the sample is substantially free of water.

[0035] As used herein the term "water insoluble solids" generally refers to the fraction of extract that is removed by centrifugation and filtration in the course of measuring water soluble solids, as described in the Examples provided herein.

[0036] As used herein, the term "pest" refers to organisms and microorganisms, including pathogens, that negatively affect plants or animals by colonizing, atacking or infecting them. This includes organisms that spread disease and / or damage the host and / or compete for host nutrients. In addition, plant pests are organisms known to associate with plants and which, as a result of that association, causes a detrimental effect on the plant's health and vigor. Plant pests include but are not limited to fungi, bacteria, insects, and nematodes.

[0037] The term "pesticide" as used herein refers to a substance that can be used in the control of agricultural, natural environmental, and domestic / household pests, such as insects, fungi, bacteria, and viruses. The term "pesticide" is understood to encompass naturally occurring or synthetic chemical insecticides (larvicides, and adulticides), insect growth regulators, acaricides (miticides), nematicides, ectoparasiticides, bactericides, fungicides, and herbicides (substance which can be used in agriculture to control or modify plant growth).

[0038] The term "plant" as used herein encompasses whole plants and parts of plants such as roots, stems, leaves and seed, as well as cells and tissues within the plants or plant parts. Target crops to be protected using the disclosed compositions and methods include, without limitation, the following species of plants: cereals (wheat, barley, rye, oats, rice, sorghum and related crops), beet (sugar beet and fodder beet), forage grasses (orchardgrass,fescue, and the like), drupes, pomes and soft fruit (apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries and blackberries), leguminous plants (beans, lentils, peas, soybeans), oil plants (rape, mustard, poppy, olives, sunflowers, coconuts, castor oil plants, cocoa beans, groundnuts), cucumber plants (cucumber, marrows, melons) fiber plants (cotton, flax, hemp, jute), citrus fruit (oranges, lemons, grapefruit, mandarins), vegetables (spinach, lettuce, asparagus, cabbages and other Brassicae, onions, tomatoes, potatoes, paprika), lauraceae (avocados, carrots, cinnamon, camphor), deciduous trees and conifers (e.g. linden-trees, yew-trees, oak-trees, alders, poplars, birch-trees, firs, larches, pines), or plants such as maize, tobacco, nuts, coffee, sugar cane, tea, vines, hops, bananas and natural rubber plants, as well as ornamentals (including composites).

[0039] The terms "control" or "controlling" used throughout the specification and claims, are meant to include any pesticidal (killing) or pestistatic (inhibiting, maiming or generally interfering) activities of a pesticidal composition against a given pest. Thus, these terms not only include killing, but also include such activities as those of chemisterilants which produce sterility in insects by preventing the production of ova or sperm, by causing death of sperm or ova, or by producing severe injury to the genetic material of sperm or ova, so that the larvae that are produced do not develop into mature progeny. The terms also include repellant activity that protect animals, plants or products from insect attack by making food or living conditions unattractive or offensive to pests. These repellant activities may be the result of repellents that are poisonous, mildly toxic, or non-poisonous to pests.

[0040] The term “substantially free of or “substantially free from” as used herein refers to either the complete absence of an ingredient or a minimal amount thereof merely as impurity or unintended byproduct of another ingredient. A composition that is “substantially free” of / from a component means that the composition comprises less than about 0.5%, 0.25%, 0.1%, 0.05%, or 0.01%, or even 0%. by weight of the composition, of the component.

[0041] Unless otherwise noted, all component or composition levels are in reference to the active portion of that component or composition, and are exclusive of impurities, for example, residual solvents or by-products, which may be present in commercially available sources of such components or compositions.

[0042] All percentages and ratios are calculated by weight unless otherwise indicated. All percentages and ratios are calculated based on the total composition unless otherwise indicated. It should be understood that every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this specification will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this specification will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein. For example, a stated range of “1 to 10” should be considered to include any and all subranges between (and inclusive of) the minimum value of 1 and the maximum value of 10; that is, all subranges beginning with a minimum value of 1 or more, e.g. 1 to 6. 1, and ending with a maximum value of 10 or less, e.g., 5.5 to 10.%DETAILED DESCRIPTION

[0043] Described herein are agricultural compositions comprising one or more renewable components. As used herein, the phrase “agricultural composition” includes compositions and formulations designed as biochemical pesticides. Such compositions may be administered to a plant to effectively treat (e.g., preventing, controlling, impeding, inhibiting, and the like) the growth or presence of fungal and bacterial organisms, pathogens, viral, microbial, and other diseases causing pests. The agricultural compositions may be in the form of a liquid, a powder, a granule or a flake and other suitable forms that may be apparent to one skilled in the art in view of the teachings herein. The agricultural compositions may comprise an extract derived non-w oody plants of the genus Hesperaloe, for example, Hesperaloe funifera, Hesperaloe nocturna, Hesperaloe parviflora, and Hesperaloe chiangii.Compositions

[0044] The compositions disclosed herein may comprise from about 1 wt. % to about 100 wt. % renewable components. For example, the compositions may comprise about 1. 5, 10, 15. 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75. 80. 85. 90. 95, or 100 wt. % renewablecomponents. The agricultural compositions described herein may comprise a renewable component derived from Hesperaloe biomass, particular an extract prepared from Hesperaloe biomass and more particularly a water-soluble extract prepared from Hesperaloe biomass, such as a crude Hesperaloe extract or a saponin-enriched extract. The Hesperaloe extract may comprise a crude extract, a processed extract, a preserved extract, or combinations thereof. The Hesperaloe extract may comprise saponins and may be included in the agricultural composition such that the composition may comprise at least about 0.5% saponin to lees than about 50 % saponin derived from Hesperaloe (e.g.. from about 1 to 50 wt. %, 2 to 25 wt. %, or 3 to 10 wt. %). For example, the agricultural compositions may comprise about 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 wt. % saponin. The agricultural compositions may comprise up to about 25, 30, 35, 40, 45, or 50 wt. % saponin. In some examples, the agricultural composition may be substantially free from saponins derived from sources other than Hesperaloe biomass. In this manner, the saponins in the agricultural composition may be renew able saponins.

[0045] In some examples, the agricultural compositions may comprise one or more saponins extracted from a non-woody plant of the genus Hesperaloe, where the one or more saponins may comprise 25(27)-dehydrofucreastatin (FIG. 2A), 5(6), 25(27)- disdehydroyuccaloiside C (FIG. 2B), 5(6)-disdehydroyuccaloiside C (FIG. 2C), furcreastatin, yuccaloiside C, or combinations thereof.

[0046] The Hesperaloe extracts of the present disclosure may provide for effective control of (microorganisms) insects, mites, fungi and microorganisms. In particular instances, such as where an insect damages a plant part or tissue and a secondary fungal disease develops, this aspect of the disclosure is particularly advantageous. The compositions described herein may have ver7good fungicidal properties and can be employed for controlling phytopathogenic fungi, such as, without limitation, plasmodiophoromycetes, oomycetes. chytridiomycetes, zygomycetes, ascomycetes, basidiomycetes. deuteromycetes, etc. Fungal phytopathogens particularly associated with crop plants and included within the scope of the present invention include, without limitation, the following: Miscellaneous Fungal Diseases (e.g., Septoria tritici, Septoria nodorum); Gibberella ear mold (e.g., e.g., Gibberella zeae, G. saubinetti); Aspergillus ear rot (e.g., Aspergillus flavus, A. parasiticus);Diplodia ear rot (e.g., Diplodia may dis, D. macrospora); Fusarium ear rot (e.g., Fusarium moniliforme, F. monilif. var. subglutinans); Pythium stalk rot (e.g., Pythium aphanidermata); Anthracnose stalk rot (e.g., Colletotrichum graminicola, C. tucumanensis, Glomerella graminicola); Diplodia stalk rot (e.g., Diplodia may dis, D. zeae-maydis, Stenocarpella may dis, Macrodiplodia zeae, Sphaeria maydis, S. zeae, D. macrospora); Fusarium stalk rot (e.g., Fusarium moniliforme); Gibberella stalk rot (e.g., G. zeae, G. saubinetti); Stewart's wilt & leaf blight (e.g., Erwinia stewartii); Northern com leaf blight (e.g., Exserohilum turcicum); Southern com leaf blight (e.g.. Bipolaris maydis); Gray leaf spot (e.g., Cercospora zeae- maydis, C. sorghi var. maydis); Anthracnose leaf blight (e.g., Colletotrichum graminicola); Common mst (e.g., Puccinia sorghi, P. maydis); Southern rust (e.g., Puccinia polysora, Dicaeoma polysorum); Head smut (e.g., Sphacelotheca reiliana); Common smut (e.g., Ustilago maydis); Carbonum leaf spot (e.g., Helminthosporium carbonum); Eye spot (e.g.. Kabatiella zeae); Sorghum downy mildew (e.g., Peronosclerospora sorghi); Brown stripe downy mildew (e.g., Sclerophthora rayssiae); Sugarcane downy mildew (e.g., Peronosclerospora sacchari); Phillipine downy mildew (e.g., Peronoscler. Philippinensis); Java downy mildew (e.g., Peronosclerospora maydis); Spontaneum downy mildew (e.g., Peronosclerospora spantanea); Rajasthan downy mildew (e.g., Peronosclerospora heteropogoni); Graminicola downy mildew (e.g., Sclerospora graminicola); Rusts (e.g., Puccinia graminis f.sp. tritici, Puccinia recondi ta f.sp. tritici, Puccinia striiformis); Smuts (e.g., Tilletia tritici, Tilletia controversa, Tilletia indica, Ustilago tritici, Urocystis tritici); Root rots, Foot rots and Blights (e.g., Gaeumannomyces Graminis, Pythium Spp., Fusarium culmorum, Fusarium graminaerum. Fusarium avenaceum, Drechslere tritici-repentis, Rhizoctonia spp., Colletotrichum graminicola, Helminthosporium spp., Microdochium nivale, Pseudocercosporella herpotrichoides); Mildews (e.g., Erysiphe graminis f.sp. tritici, Sclerophthora macrospora), and the like.

[0047] The compositions described herein may be useful in the treatment of grampositive cocci that cause staphylococcal infections such as pneumonia, bacteremia, osteomyelitis, enterocolitis, and the like; streptococci that cause infections such as hemolytic, viridans, enterococci, lactic, and the like; pneumococci that cause infections such as pneumonia, sinusitis, otitis, Meningitis, and the like; gram-negative cocci such asmeningococcus, gonococcus, and the like; gram-positive bacilli that cause infections such as erysipelothricosis, listeriosis, anthrax, nocardiosis, and the like; gram-negative bacilli that cause infections such as enterobacteriaceac salmonella, shigellosis, hemophilus, tularemia, plaque, melioidosis, bartonellosis. Campylobacter, and noncholera vibrio, and the like; anaerobic bacilli that cause infections such as Clostridium botulinum, Clostridium tetany, clostridia of gas gangrene bacteroides, mixed anaerobic, actinomycosis, and the like; mycobacteria that cause infections such as tuberculosis and leprosy, and the like; and spirochetes that cause diseases such as leptospirosis, lyme disease, and endemic treponematoses. Further, the present invention, the pesticidal compositions may be useful for treating surfaces containing infectious human immunodeficiency virus (HIV), influenza, A, B, and C, parainfluenza viruses 1-4, rhono viruses (common cold), mumps virus, adenoviruses, reoviruses, and epstein-Barr virus, infants and adult syncytial vims, primary atypical pneumonia, polioviruses, coxsackieviruses, echoviruses and high numbered viruses, epidemic gastroenteritis viruses, rubeola virus, rubella virus, varicella-zoster virus, herpes simplex, human herpes vims ty pe 6, human parvovirus Bl 9, cytomegalovirus, hepatitis viruses types A, B. C, D. human Papillomavirus, molluscum contagiosum virus, arboviruses, togaviruses, alphaviruses, flaviviruses, bunyaviruses, orbivirus. rabies virus, herpesvirus simiae, arenaviruses, filovimses, and the like.

[0048] The agricultural compositions may be applied or administered by any one of a number of well-known methods and may vary depending on the locus of the fungal infection or potential fungal infection. The compositions may be applied to surfaces of plant foliage, flowers, seeds, fruits and vegetables, roots, tubers, and even the soil in the vicinity of seeds, plants, and the like.

[0049] In certain instances, the compositions comprise a suitable solid or liquid carrier or vehicle that is compatible with delivery to a plant being treated. The carrier may be present in an amount of about 50 to about 99 wt. % of the composition. For example, the composition of the present disclosure may be formulated as an aqueous spray or dip, wettable powder, drench, dust, granule, pellet, or the like, when applying the compositions to plants. Where the composition comprises a liquid carrier the concentration of Hesperctloe extract may be at least about 100 ppm, at least about 500 ppm, at least about 1000 ppm, at least about1500 ppm, at least about 2000 ppm, at least about 2500 ppm, at least about 3000 ppm, at least about 3500 ppm, at least about 4000 ppm, at least about 4500 ppm, at least about 5000 ppm, such as from about 100 to about 6000 ppm.

[0050] The agricultural compositions of the present disclosure generally comprise a quantity of Hesperaloe derived saponins sufficient to obtain a reduction in the level of disease. For example, the compositions may comprise a relatively modest amount of saponins, such as less than about 50 wt. %, such as less than about 30 wt. %, such as less than about 25 wt. %, such as less than about 20 wt. %, such as from about 1.0 wt. % to about 50 wt. %.

[0051] The agricultural compositions may also, optionally, comprise excipients, lubricants, binders, disintegrators for solid pharmaceutical preparations; and solvents, solubilizing agents, suspending agents, isotonizing agents, or buffering agents. In addition, additives such as ordinary preservatives, antioxidants, colorants and wetting agents may be added as necessary.

[0052] Formulations containing the agricultural compositions may be prepared in any known manner, for instance by extending the pesticidal compositions with conventional liquid carriers and / or dispersible solid carriers optionally with the use of carrier vehicle assistants, e g. conventional pesticide surface-active agents, including emulsifying agents and / or dispersing agents, whereby, for example, in the case where w ater is used as diluent, organic solvents may be added as auxiliary' solvents. Suitable liquid diluents or carriers include water, petroleum distillates, or other liquid carriers with or without surface active agents. The choice of dispersing and emulsifying agents and the amount employed is dictated by the nature of the composition and the ability of the agent to facilitate the dispersion of the pesticidal compositions of the present invention. Non-ionic, anionic, amphoteric, or cationic dispersing and emulsifying agents may be employed, for example, the condensation products of alkylene oxides with phenol and organic acids, alkyl aryl sulfonates, complex ether alcohols, quarternary' ammonium compounds, and the like.

[0053] Liquid concentrates may be prepared by dissolving a composition of the present disclosure with a solvent and dispersing the agricultural compositions inventions in water with suitable surface active emulsifying and dispersing agents.

[0054] In certain embodiments the agricultural compositions may be provided as solids, such a granules, intended for topical application to a plant. The solid compositions may comprise from about 1.0 to about 50 wt. % extract pesti ci daily-effective amount. Thus, the contemplated are formulations with solid carriers or diluents such as bentonite, fullers earth, calcium carbonate, ground natural minerals, such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, vermiculite, and ground synthetic minerals, such as highly-dispersed silicic acid, alumina and silicates, crushed and fractionated natural rocks such as calcite, marble, pumice, sepiolite and dolomite, as well as synthetic granules of inorganic and organic meals, and granules of organic materials such as sawdust, peanuts, apple pomace, mustard, lecithin, recycled paper, coconut shells, com cobs, and tobacco stalks. Adhesives, such as carboxymethyl cellulose, natural and synthetic polymers, (such as gum arabic, polyvinyl alcohol and polyvinyl acetate), and the like, may also be used in the formulations in the form of powders, granules or emulsifiable concentrations.

[0055] In some examples, the composition may be substantially free of living microorganisms. In certain examples, the composition may comprise no living microorganisms. In some examples, the composition may be substantially free from geologically derived components.

[0056] In some examples, the composition descirbed herein may be a composition for treating and / or preventing a bacterial plant pathogen. The composition may comprise an extract of a non-woody plant of the genus Hesperaloe and a carrier. The extract may comprise at least one saponin and be present in an amount effective to inhibit the growth or presence of fungal and bacterial organisms, pathogens, viral, microbial, and other diseases causing pests.

[0057] In some examples, the composition may be a pesticide. The pesticide may be a biochemical pesticide or biopesticide, i.e., the composition comprises only naturally occurring substances and damages, kills, or repels organisms that are considered as pests.

[0058] In some examples, the pesticide may comprise an extract of a non-woody plant of the genus Hesperaloe, including Hesperaloe fiinifera, Hesperaloe noctiirna, Hesperaloe parviflora, Hesperaloe chiangii, or a combination thereof. The extract may comprise at least one saponin, and the at least one saponin may be present in an amount less than 50 wt. %. Forexample, the at least one saponin may be present in an amount from about 1 wt. % to 40 wt. %, about 5 to 30 wt. %, or about 10 to 25 wt. %. The extract may further comprise a lipid, a protein, an inorganic salt, a saccharide, a polysaccharide, an organic acid, or combinations thereof.

[0059] In some examples, the pesticide may further comprise a carrier comprising bentonite, calcium carbonate, cellulose, clove, dolomite, kaolin, lecithin, mustard, water, or combinations thereof. The carrier may be present in an amount from about 50 to about 99 wt. %.

[0060] The composition of the Heserpaloe extract may vary depending on the source of the material and the purification process, how ever, in certain embodiments it may be preferable that the extract retain one or more natural constituents in addition to saponins, such as lipids, proteins, inorganic salts, saccharides, polysaccharides and / or organic acids. For example, the extract may comprise the constituents set forth in Table 1. Each constituent in Table 1 is provided on wt. % based upon the as is bone-dry extract.Table 1Methods of Making and Use

[0061] Saponins may be derived from non-w oody plants of the genus Hesperaloe, for example, Hesperaloe fiinifera, Hesperaloe nocturna, Hesperaloe parviflora, and Hesperaloe chiangii. The saponins may be extracted from Hesperaloe biomass by grinding, cutting, pressing, crushing, and / or mechanical abrasion (e.g., milling, depithing, or a combination thereof) of the biomass in combination with solvent extraction. In some examples, the biomass may be subjected to compression to remove intercellular and intracellular liquid. Compression may be carried out using one or more opposed counter-rotating rolls, a mechanical press, a screw press or direct hydraulic pressure.

[0062] The initial extract may be a crude extract, which may include, for example, fats, proteins, saccharides, polysaccharides, saponins, sapogenins, minerals (also referred to as inorganic salts), and organic acids. The extracted substances may also include waterinsoluble components that may be suspended in the Hesperaloe crude extract. The crude extract may be subjected to further processing to produce a Hesperaloe extract with increased saponin concentration (also referred to herein as Hesperaloe saponin-enriched extract).

[0063] FIG. 3 illustrates an example multistage process 300 of the present disclosure. In the illustrated multistage process 300, a mill and press 302 is used to produce a Hesperaloe crude extract (juice) 304 from Hesperaloe biomass 301 . The crude extract 304 is purified to remove at least a portion of the fats, proteins, saccharides, polysaccharides, and / or waterinsoluble components by heat and / or centrifuge 314. The resulting Hesperaloe processed extract (clarified juice) 316 is further purified by centrifuge, filtration or heat 320 to produce a Hesperaloe saponin-enriched extract (syrup) 322. In some examples, the extract 316 may be filtered using one or more membranes having a pore size sufficient to allow saponin retention, which is described in more detail herein.

[0064] The Hesperaloe crude extract 304 may be produced, for example, by milling (and optionally imbibition and / or depithing) Hesperaloe biomass in the presence of a solvent. Generally, processing Hesperaloe biomass removes at least about 25% of the water-soluble solids from the biomass, more preferably at least about 50%, still more preferably at least about 75%. such as from about 25% to about 98%, such as from about 50% to about 90%, such as from about 75% to about 90%. Examples of useful solvents may include, but are not limited to, acetic acid, acetone, acetonitrile, benzene, 1 -butanol, 2-butanol, 2-butanone, t- butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, 1,2- dichloroethane, diethylene glycol, diethyl ether, diglyme, 1 ,2-dimethoxy ethane, dimethylformamide, dimethylsulfoxide, 1,4-dioxane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide, hexamethylphosphorous triamide, hexane, methanol, methyl-t-butyl ether, methylene chloride, N-methyl-2-pyrrolidinone, pentane, perchloroethylene, petroleum ether, 1 -propanol, 2-propanol, pyridine, tetrahydrofuran, toluene, tri ethyl amine, trifluorotoluene, water, xylene, the like, and any combination thereof.

[0065] In one example, the process for producing aHesperaloe crude extract 304 may comprise contacting aHesperaloe biomass with a solvent comprising water and separating the water-soluble fraction from the insoluble biomass fraction. In another example, the extractant solution may comprise, in addition to water, a surfactant, another solvent, and / or Hesperaloe crude extract. The Hesperaloe crude extract can come from, for example, an earlier extraction step or an earlier milling step. In yet another example, aHesperaloe crude extract 304 may also be produced with a first extraction using acetone or diethyl ether to remove lipids and pigments followed by a second extraction using methanol as a solvent, or a mixture of methanol and water. In another example, the solvent may be a 4: 1 ethanol-water solvent.

[0066] Additional details of milling and other processes used to produce the Hesperaloe crude extract 304 produced from the Hesperaloe biomass are described in US Patent App. Pub. No. 2023 / 0233644, which is incorporated herein by reference.

[0067] The amount of water-insoluble solids recovered from a biomass may vary depending on the extraction efficiency; however, in certain instances from about 1 grams to about 100 grams of water-soluble solids may be extracted per kilogram of bone-dry biomass, such as from about 1 grams to about 30 grams per kilogram, such as from about 10 grams to about 50 grams per kilogram.

[0068] The amount of water-soluble solids recovered from a biomass may vary depending on the extraction efficiency; however, in certain instances from about 100 grams to about 400 grams of water-soluble solids may be extracted per kilogram of bone-dry biomass, such as from about 120 grams to about 350 grams per kilogram, such as from about 150 grams to about 300 grams per kilogram.

[0069] A Hesperaloe crude extract 304 may have a water-soluble solids concentration of about 5% to about 90%, such as about 5% to about 30%, such as about 25% to about 60%. such as about 50% to about 90%, based on a total weight of the Hesperaloe crude extract. The Hesperaloe crude extract may comprise saccharides and polysaccharides at a concentration of at least about 1 wt. %, such as from about 1 wt. % to about 15 wt. %, such as from about 2 wt. % to about 10 wt. %, based upon the total weight of water-soluble solids. The Hesperaloe crude extract may comprise saponins at a concentration of at least about 0.5wt. %, such as at least 1 wt. %, such as at least 5 wt. %, such as from about 0.5 wt. % to about 40 wt %, such as from about 1 wt. % to about 20 wt. %, such as from about 5 wt. % to about 30 wt. %, based upon the total weight of water-soluble solids. Additionally and / or alternatively, the Hesperaloe crude extract may comprise sapogemns at a concentration of at least about 0.5 wt. %, such as at least 1 wt. %, such as at least 5 wt. %, such as from about 0.5 wt. % to about 40 wt. %, such as from about 1 wt. % to about 20 wt. %, such as from about 5 wt. % to about 30 wt. %, based upon the total weight of w ater-soluble solids. The Hesperaloe crude extract may comprise proteins at a concentration of at least about 1 wt. %, such as from about 1 wt. % to about 30 wt. %, such as from about 5 wt. % to about 25 wt. %, based upon the total weight of w ater-soluble solids. The Hesperaloe crude extract may comprise fats at a concentration of about 0 wt. % to about 25 wt. %, such as about 1 wt. % to about 20 wt. %, such as about 5 wt. % to about 15 wt. %, based upon the total weight of water-soluble solids. The Hesperaloe crude extract may comprise inorganic salts at a concentration of about 5 wt. % to about 35 wt. %, such as about 5 wt. % to about 30 wt. %, such as about 7 wt. % to about 20 wt. %, based upon the total weight of w ater-soluble solids.

[0070] The Hesperaloe crude extract 304 may be purified by one or more treatment steps 314 to remove at least a portion of the fats, proteins, saccharides, polysaccharides, and / or water-insoluble components and produce the Hesperaloe processed extract 316. Examples of treatments may include, but are not limited to, flocculation, centrifugation, foam fractionation, solvent extraction, the like, and any combination thereof. For example, the Hesperaloe crude extract 304 may be purified by flocculation followed by centrifugation 314 to facilitate removal of the flocculants. In this example, the resulting supernatant is the Hesperaloe processed extract 316. Alternatively, additional treatments (e.g., solvent extraction or foam fractionation) may be performed on the supernatant to produce the Hesperaloe processed extract 316.

[0071] Centrifugation may be used to remove higher density substances from the Hesperaloe processed extract and, when used in combination with flocculation, to remove flocculants from the Hesperaloe processed extract. Suitable rpm to achieve a desired g-force that removes the foregoing may be determined by one of ordinary' skill in the art without undue experimentation. After centrifugation, the supernatant may be the Hesperaloeprocessed extract, or the supernatant may be further purified to produce the Hesperaloe processed extract. In some examples, centrifugation and flocculation followed by centrifugation may be preferred to minimize the amount of solvent needed to remove at least a portion of the fats, proteins, saccharides, polysaccharides, and / or water-insoluble components.

[0072] When centrifugation follows flocculation, an amount of time between initiating flocculation and starting centrifugation may be about 30 minutes or less, such as about 15 minutes or less, such as about 5 minutes or less, such as about 1 minute or less, such as about 10 seconds or less to about 30 minutes, such as about 10 seconds to about 15 minutes, such as about 10 second to about 5 minutes.

[0073] The amount of water-soluble solids recovered from the Hesperaloe crude extract will vary depending on the treatment and its efficiency. The Hesperaloe processed extract may have a water-soluble solids concentration of from about 5% to about 90%, such as from about 5% to about 30%, from about 25% to about 60%, or from about 50% to about 90%, based on a total w eight of the Hesperaloe processed extract. The Hesperaloe processed extract may comprise saccharides and polysaccharides at a concentration of at least about 1 wt. %, such as from about 1 wt. % to about 15 wt. %, from about 2 wt. % to about 10 wt. %. based upon the total weight of w ater-soluble solids. The Hesperaloe processed extract may comprise saponins at a concentration of at least about 0.5 wt. %, such as at least 1 wt. %, at least 5 wt. %, from about 0.5 wt. % to about 40 wt. %, from about 1 wt. % to about 20 wt. %, or from about 5 wt. % to about 60 wt. %, based upon the total weight of water-soluble solids. The Hesperaloe processed extract may comprise sapogenins at a concentration of at least about 0.5 wt. %, such as at least 1 wt. %, at least 5 wt. %, from about 0.5 wt. % to about 40 wt. %, from about 1 wt. % to about 20 wt. %, or from about 5 wt. % to about 60 wt. %, based upon the total weight of water-soluble solids. The Hesperaloe processed extract may comprise proteins at a concentration of at least about 1 wt. %. such as from about 1 wt. % to about 20 wt. %, or from about 5 wt. % to about 15 wt. %, based upon the total weight of water-soluble solids. The Hesperaloe processed extract may comprise fats at a concentration of from about 0 wt. % to about 20 wt. %, such as from about 1 wt. % to about 15 wt. %, or from about 3 wt. % to about 10 wt. %. based upon the total weight of water-soluble solids.The Hesperaloe processed extract may comprise inorganic salts at a concentration of from about 5 wt. % to about 35 wt %, such as from about 5 wt. % to about 30 wt. %, or from about 7 wt. % to about 20 wt. %, based upon the total weight of water-soluble solids. For example, the Hesperaloe processed extract may comprise at least about 1 wt. % saccharides and polysaccharides, at least about 0.5 wt. % saponins, at least about 0.5 wt. % sapogenins, at least about 1 wt. % proteins, from about 0 wt. % to about 20 wt. % fats, and from about 5 wt. % to about 35 wt. % inorganic salts, each based upon a total weight of water-soluble solids of the Hesperaloe processed extract.

[0074] The Hesperaloe processed extract may be filtered through one or more membranes 320 to produce a Hesperaloe saponin-enriched extract 322. The one or more membranes includes at least one membrane with a pore size sufficient to enrich a saponin concentration on a dry basis in a retentate relative to a saponin concentration on a dry’ basis in the Hesperaloe processed extract. Stated another way, at least one of the membranes has a pore size sufficient to allow saponin retention. It should be noted that 100% saponin retention is not implied, but saponin retention is preferably at least 50 wt. %, and more preferably at least 80 wt. %. Said membrane may have a pore size characterized by a distance measurement (e.g.. nm or pm) or a nominal molecular weight limit (NMWL). For example, the membrane that allows saponin retention may have a pore size of about 5 nm or less, such as from about 0.5 nm to about 5 nm, or from about 0.5 nm to about 3 nm. If the pore size is characterized by NWML, the membrane that allows saponin retention may have a pore size (or NMWL) of about 600 Da or less, such as from about 100 Da to about 600 Da, from about 100 Da to about 500 Da, or from about 300 Da to about 500 Da.

[0075] In some examples, it may be preferable to minimally process the crude Hesperaloe extract to minimize costs and maintain various metabolites, including alkaloids, phenols, terpenoids, and secondary’ substances developed by plants to help them in the protection from pests. Such minimal processes may be targeted to remove suspended solids, also referred to herein as the water insoluble fraction, and may involve any number of well- known processes including, for example, clarification, filtration, centrifugation, or a combination thereof. In certain instances, it may be preferable to remove at least about 90% water insoluble solids from the crude extract, such as at least about 95%, such as at leastabout 98%. Generally, the water insoluble solids comprise hydrophobic substances such as waxes and pectins. In some embodiments, the clarified extract is substantially free of suspended solids. After removal of suspended solids, the clarified extract may be used directly or concentrated.

[0076] In certain examples, the suspended solids may be removed from the crude extract by mixing the crude extract with a solvent, and optionally a salt, to form a first solution. The solvent may comprise one or more solvents selected from acetic acid, acetone, acetonitrile, benzene, 1 -butanol, 2- butanol, 2-butanone, t-butyl alcohol, carbon tetrachloride. chlorobenzene, chloroform, cyclohexane, 1,2-dichloroethane, di ethylene glycol, diethyl ether, diglyme, 1,2-dimethoxy ethane, dimethylformamide, dimethyl sulfoxide, 1,4-di oxane, ethanol, ethyl acetate, ethylene glycol, glycerin, heptane, hexamethylphosphoramide, hexamethylphosphorous triamide, hexane, methanol, methyl-t-butyl ether, methylene chloride, N-methyl-2-pyrrolidinone, pentane, perchloroethylene, petroleum ether, 1 -propanol, 2-propanol, pyridine, tetrahydrofuran, toluene, triethylamine, trifluorotoluene, water, xylene, or any combination of the forgoing. In some embodiments the solvent is w ater. In those instances where a salt is added, the salt may be selected from an alkali metal salt, an alkaline earth salt, a transition metal salt, an ammonium salt, or combinations of the forgoing. In certain preferred embodiment the salt added to the plant extract to form the solution is an alkaline earth metal salt. In particularly preferred embodiments the salt is calcium chloride (CaCb), magnesium chloride (MgCh), or a mixture thereof.

[0077] The pH of the first solution may be adjusted such that the pH ranges from about 6.0 to about 9.0, such as from about 6.0 to about 8.0, such as from about 6.0 to about 7.0.

[0078] In some examples, at least one phosphate may be added to the first solution described above to form an ion-polysaccharides complex precipitate. Useful phosphates include, for example, sodium hydrogen phosphate (Na2HPO4), sodium dihydrogen phosphate (NaH2PO4), sodium phosphate (NasPC ), or sodium hydrogen bisphosphate (Na2H2PO?). The precipitated ion- polysaccharides complex may be removed by filtration to yield a second solution, which may be further clarified to produce a saponin enriched extract. Optionally, the extract can be concentrated by any filtration technique known in the prior art such as nanofiltration, ultrafiltration and diafiltration, or any combination of these techniques.

[0079] Also disclosed are methods of making and using agricultural compositions formulated for application to a plant. Thus, in certain embodiments, the method may further comprise combining a Hesperaloe extract with a carrier. The extract may comprise at least one saponin. The extract may further comprise one or more of a lipid, a protein, an inorganic salt, a saccharide, a polysaccharide and / or an organic acid. The method may further comprise adding an additive to the mixture.

[0080] Also disclosed are methods of making and using the agricultural compositions described herein. A method of making an agricultural composition is shown in FIG. 4. The method 400 may comprise combining aHesperaloe extract with a carrier 410; and optionally, adding a surfactant 420 or additive 430 to the mixture of extract and carrier to produce the agricultural composition 440. A method of using the agricultural compositions described herein may comprise contacting at least a portion of a targeted substrate with the agricultural composition. Example substrates include but are not limited to plants, crops, and wood. The composition may be formulated as a liquid and applied by spraying, dipping, or drenching the substrate. The composition may be formulated as a gel or solid and placed in contact with the targeted substrate. Examples of solid formulation include but are not limited to a wettable powder, a dust, a crystal, a granule, a pellet, and the like. In some examples, method may comprise impregnating a material, such as posts, with the agricultural composition. The agricultural compositions may be applied to a broad range of crops to inhibit or protect against a wide range of pests by applying a pesticidally-effective amount of the composition to a location or substrate where pest control is desired.EXAMPLES

[0081] The following non-limiting examples illustrate the present disclosure. It should be noted that various changes and modifications can be applied to the following examples and processes without departing from the scope of this disclosure, which is defined in the appended claims. Therefore, it should be noted that the following examples should be interpreted as illustrative only and not limiting in any sense.Example 1: Method for Determining Water Soluble Solids

[0082] Total biomass water soluble solids was determined using an Accelerated Solvent Extraction system (ASE) (Dionex TM ASE™ 350 (Thermo Fisher Scientific, Waltham, MA)). Approximately 10 grams of harvested biomass was dried to a constant weight in an oven, typically 4 hours at 125 °C. After drying 1.5 - 2.0 grams of the bone dry biomass was accurately weighed and the weight (Wb) recorded to the nearest 0.001 gram. Using water as the solvent, biomass was extracted using the conditions set forth in Table 2. The ratio of biomass to solvent was generally 21: 1 and five consecutive water extraction cycles were performed. At the end of each extraction cycle, the liquid phase was collected, dried under vacuum at approximately 40 °C and the weight of the dried material (Wi) was recorded to the nearest 0.001 g. The total weight of water soluble solids (We) was calculated by summing the weight of solids recovered from each extraction cycle (Wi). Total water soluble solids as a percentage of bone dry biomass was then determined using the following equation: Water Soluble Solids (wt. %)= We / Wb *100.Table 2

[0083] The total water soluble solids in biomass extract was determined by withdrawing an appropriate aliquot, about 10-50 mL, transferring to clean, dry, centrifuge tube. The tube was centrifuged at 7000 rpm for 20 minutes. The weight of extract (WI) was calculated. An aliquot of the supernatant was transferred to clean, pre-weighed beaker (Do), and weighed. The beaker and sample were weighed to the nearest 0.001 g and the weight (D2) recorded. The beaker containing the sample was placed at 140 °C in a hot air oven for overnight drying. The beaker was removed from the oven and desiccated to cool to room temperature thenweighed to the nearest 0.001 gram (DI). The weight percentage of soluble solids, based upon the weight of the extract, was determined using the formula below:(DI — Do)x 100Dl = mass of empty beaker+ dried soluble solids, Do = mass of empty beaker, D2 = mass of biomass extract and empty beaker.Example 2: Method for Determining Total Saponins

[0084] Total saponins were measured generally as described in Makkar, Harinder P.S.. Sidhuraju, P., Becker, Klaus (2007) Plant Secondary Metabolites, chapter 17, pp 93-100. A standard saponin solution was prepared by weighing 10 mg of diosgenin (MilliporeSigma >93%), dissolving in 16 mL of methanol and adding 4 ml of distilled water. The solution was mixed thoroughly to yield a 0.5 mg / mL diosgenin solution in 80% methanol solvent. The standard was used to produce a calibration curve by transferring various amounts of the standard (0, 10, 20, 40, 60, 80, and 100 pl) into 13-mm glass test tubes. A solution of 80% aqueous methanol was added to a total volume of 100 pL.

[0085] Prior to testing, samples of biomass extract were adjusted to about 0.5 wt. % total solids by dilution with water to ensure absorbency result fell along the saponin standard calibration curve range. Samples of diluted extract (20-pL) were pipetted into 13-mm glass test tubes and the volume was brought up to 100 pL with 80 pL methanol. Each sample was tested in triplicate.

[0086] To each sample 100 pL of vanillin reagent (prepared by dissolving 800 mg of vanillin in 10 mL of 99.5% ethanol (analytical grade)) and then 1.0 mL of 72% (v / v) sulfuric acid (72% (v / v) sulfuric acid prepared by adding 72 mL of sulfuric acid (analytical grade, 95%, w / w) to 28 mL of distilled water) were added. Solutions were mixed well and heated at 60 °C for 10 minutes. Samples w ere then cooled in an ice bath and 1 mL of solution wastransferred into respective cuvette and absorbance at 544 nm was read. The total mass of saponins in the sample was calculated based upon the standard absorbency curve as follows:Saponin (pg) = [Slope] x Measured Absorbency - [Intercept]Example 3

[0087] Fresh cut Hesperaloe funifera leaves were pressed using a mechanical juicer. A fraction of the resulting juice, which comprised 413 g (15.6%) solids, was mixed with 300 ml of methanol to form a precipitate. The precipitate was removed by centrifugation and yielded an amber color clarified juice (CLJ). A portion of the CLJ was dried at 80 °C to form a dried extract (CLJ A). The dried extract (CLJ A) was dissolved in water, filtered, and dried at 80 °C to form a second dried extract (CLJ B).

[0088] The saponin containing Hesperaloe extract was prepared by forage harvesting mature Hesperaloe funifera leaves above the crow n, cutting the leaves into pieces ranging from about 0.50 to about 8.0 cm and pressing the cut biomass using a sugar cane tandem press, each mill of the tandem press having 3 rollers. The biomass was pressed three times and the crude juice was collected and passed through 25 mm filter followed by microfiltration / nanofiltration to concentrate the extract to 15% solids. The extract was mixed with 103 mL of methanol to form a brown precipitate, which was removed by centrifugation. The brown clarified solution was dried at 80 °C to form a dried extract (CLS).

[0089] The samples were dissolved in autocleaved water to prepare 100, 1000 and 4000 ppm solutions, based upon solids. The various formulations were screened for their inhibition efficacy for different bacteria and fungus against negative and positive controls.

[0090] CLJ A, CLJB and CLS at three different levels were screened against 3 bacteria and 3 fungi that are important to different crops. The screening results are shown in Tables 3- 8. As these bacteria and fungi are slow growing, 5 days were given before results was scored. For positive drug control streptomycin was used as for bacteria, and difenconazole and trifloxystrobin w ere used for fungi. The amount of growth observed is indicated with asterisks, where the number of (*) correspond to the size of colony. E.g., (****) indicates more growth than (***), which was greater than (**), which was greater than (*). A zero (0) indicates no growth was observed.Table 3Table 4Table 5Table 6Table 7Table 8Table 9

[0091] It is well known in the industry that fungi are hard to control, shown from the results that even drugs cannot completely stop the grow th of some fungi, particularly for Pythium. However, all clarified Hesperaloe extracts at 4000 ppm were able to completely inhibit Phytophthora capsici. Using clarified Hesperaloe extracts, containing about 25 wt. % saponin as well as many other phytochemicals, the whole extract can completely inhibit the growth of bacteria Erwinia, Xanthomonas, fungi Phytophthora capsica.

Claims

We claim:

1. An agricultural composition comprising: an extract of a non-woody plant of the genus Hesperaloe, the extract comprising at least one saponin; and a carrier, wherein the saponin is present in an amount effective to inhibit the growth of a fungal or bacterial organism.

2. The composition of claim 1, wherein the at least one saponin is present in an amount less than 50 wt. %.

3. The composition of claim 1 or 2, wherein the at least one saponin is present in an amount from 1 wt. % to 50 wt. %.

4. The composition of any one of claims 1-3, wherein the extract further comprises a lipid, a protein, an inorganic salt, a saccharide, a polysaccharide, an organic acid, or combinations thereof.

5. The composition of any one of claims 1-4, wherein the non-woody plant comprises Hesperaloe funifera, Hesperaloe nocturna, Hesperaloe parviflora, Hesperaloe chiangii, or a combination thereof.

6. The composition of any one of claims 1 -5, wherein the non-woody plant is Hesperaloe funifera.

7. The composition of any one of claims 1-6. wherein the carrier comprises bentonite, calcium carbonate, cellulose, clove, dolomite, kaolin, lecithin, mustard, water, or combinations thereof.

8. The composition of any one of claims 1-7, wherein the carrier is present in an amount from about 50 to about 99 wt. %.

9. The composition of any one of claims 1-8. wherein the saponin comprises 25(27)-dehydrofucreastatin, 5(6),25(27)-disdehydroyuccaloiside C, 5(6)- disdehydroyuccaloiside C, furcreastatin, yuccaloiside C, or combinations thereof.

10. A pesticide comprising: an extract of a non-woody plant of the genus Hesperaloe, the extract comprising at least one saponin, wherein the at least one saponin is present in an amount less than 50 wt. %.

11. The pesticide of claim 10, wherein the extract further comprises a lipid, a protein, an inorganic salt, a saccharide, a polysaccharide, an organic acid, or combinations thereof.

12. The pesticide of claim 10 or 11, wherein the non-woody plant comprises Hesperaloe fumfera. Hesperaloe nocturna, Hesperaloe parviflora, Hesperaloe chiangii, or a combination thereof.

13. The pesticide of any one of claims 10-12, wherein the non-woody plant is Hesperaloe funifera.

14. The pesticide of any one of claims 10-13, wherein the at least one saponin is present in an amount from 1 wt. % to 40 wt. %.

15. The pesticide of any one of claims 10-14, further comprising a carrier comprising bentonite, calcium carbonate, cellulose, clove, dolomite, kaolin, lecithin, mustard, water, or combinations thereof.

16. The pesticide of claim 15, wherein the carrier is present in an amount from about50 to about 99 wt. %.

17. The pesticide of claim 10, wherein the pesticide is a biochemical pesticide.

18. A composition for treating and / or preventing a bacterial plant pathogen, the composition comprising: an extract of a non-woody plant of the genus Hespera 'oe. the extract comprising at least one saponin; and a carrier, wherein the saponin is present in an amount effective to inhibit the growth of a bacterial organism.

19. The composition of claim 18, wherein the extract further comprises a lipid, a protein, an inorganic salt, a saccharide, a polysaccharide, an organic acid, or combinations thereof.

20. The composition of claim 18 or 19, wherein the non-woody plant comprises Hesperaloe funifera, Hesperaloe nocturna, Hesperaloe parviflora, Hesperaloe chiangii, or a combination thereof.

21. The composition of any one of claims 18-20, wherein a concentration of extract ranges from about 1000 to about 5000 ppm.

22. The composition of any one of claims 18-21, wherein the composition is substantially free of living microorganisms.

23. The composition of any one of claims 18-22, w herein the composition is substantially free from geologically derived components.

24. A method of making the agricultural composition of any one of claims 1-9 or 18-23 and / or the pesticide of claims 10-17, the method comprising: combining the Hesperaloe extract with the carrier to form a mixture.

25. The method of claim 24, further comprising: adding optional additives and / or a surfactant to the mixture.

26. The method of claim 24 or 25, wherein the Hesperaloe extract comprises a crude extract, a processed extract, or combinations thereof.

27. A method of using the composition of any one of claims 1-9 or 18-23 and / or the pesticide of claims 10-17, the method comprising: contacting at least a portion of a targeted substrate with the composition.

28. The method of claim 27, wherein the substrate comprises a plant, a crop, wood, or combinations thereof.

Images