Patent Application
20230276801
This application claims the benefit, under 35 U.S.C. § 119 (d), of European Patent Application No. EP20382671.4, filed on Jul. 27, 2020, European Patent Application No. EP20382893.4, filed on Oct. 9, 2020, and European Patent Application No. EP20383070.8, filed on Dec. 9, 2020, the entire contents of each of which are incorporated by reference herein.
The present invention is directed to methods and compositions for increasing a growth characteristic of a plant, increasing nutrient use efficiency of a plant, or improving a plant’s ability to overcome biotic or abiotic stress comprising applying a composition comprising mixed linked β-1,3/β-1,4 glucans (MLGs) to a plant and/or part thereof, and/or growth media.
Plants have evolved a complex immune system comprised by several defense layers. One of these layers is known as Pattern-Triggered Immunity (PTI) and is based in the recognition by plasma membrane-resident Pattern Recognition Receptors (PRRs) of Damage/Microbe-Associated Molecular Patterns (DAMPs/MAMPs). PTI responses feed into protein kinase signaling cascades, ending-up in gene reprogramming processes that may ultimately allow plant surveillance to pathogen/pest attack. The relevance of PTI is well-illustrated by the fact that immune responses and disease resistance to pathogens are compromised in plants defective in PRRs perceiving DAMPs/MAMPs. Examples of MAMPs that induce PTI in plants are eubacterial flagellin and peptidoglycan, lipopolysaccharides (LPS) from gram negative bacteria or fungal cell wall-derived glucans, chitins, mannans and proteins.
Glucans represent a group of widely distributed polysaccharides mainly in the extracellular layers of numerous phylogenetic groups across the tree of life. These include a wide variety of structures, mainly with β -linkages, although α -linked glucans also occur in many species. Mixed-linked glucans [MLGs; β-1,3/1,4-glucans; (1,3;1,4)-β-D-glucans] consist of unbranched and unsubstituted chains of β-1,4-glucosyl residues discontinued by β-1,3-linkages. MLGs are widely distributed as matrix polysaccharides in cell walls of the Poaceae, but have been also reported in other groups such as in the cell walls of Equisetum spp. and other vascular plants outside of the Poaceae, as well as bryophytes and algae, lichen-forming ascomycete symbionts and fungi. β-Glucans are well-known modulators of the immune system in mammals, but little is known about their roles in plants.
There is socio-economical pressure to provide alternatives that replace or reduce the use of agrochemicals in agriculture with a focus on the development of natural products, for a more environment-friendly and sustainable solution. The present invention overcomes the shortcomings in the art by providing novel composition and methods for use in agriculture that are more environmentally friendly.
The following embodiments and aspects thereof are described and illustrated in conjunction with products and methods, which are meant to be exemplary and illustrative, not limiting in scope.
The present disclosure has applications in the agronomic sector. One embodiment provides methods and compositions for increasing a growth characteristic of a plant comprising applying a composition comprising MLGs to a plant and/or part thereof and/or to the media in which a plant and/or part thereof is growing.
Another embodiment provides increasing nutrient use efficiency of a plant comprising applying a composition comprising MLGs to a plant and/or part thereof and/or to the media in which a plant and/or part thereof is growing.
An additional embodiment provides increasing biotic and/or abiotic stress tolerance or resistance in a plant and/or part thereof comprising applying a composition comprising MLGs to a plant and/or part thereof and/or to the media in which a plant and/or part thereof is growing.
In some embodiments, a composition comprising MLGs may further comprise, a surfactant, a humectant, an adjuvant, an antioxidant, a preservative, a plant macronutrient, a plant micronutrient, a plant growth regulator, a pesticide, a fungicide, an antiviral, an anti-bacterial, and/or an herbicide.
These and other aspects of the invention are set forth in more detail in the description of the invention below.
The present invention now will be described hereinafter with reference to the accompanying drawings and examples, in which embodiments of the invention are shown. This description is not intended to be a detailed catalog of all the different ways in which the invention may be implemented, or all the features that may be added to the instant invention. For example, features illustrated with respect to one embodiment may be incorporated into other embodiments, and features illustrated with respect to a particular embodiment may be deleted from that embodiment. Thus, the invention contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted. In addition, numerous variations and additions to the various embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure, which do not depart from the instant invention. Hence, the following descriptions are intended to illustrate some particular embodiments of the invention, and not to exhaustively specify all permutations, combinations, and variations thereof.
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.
All publications, patent applications, patents and other references cited herein are incorporated by reference in their entireties for the teachings relevant to the sentence and/or paragraph in which the reference is presented.
Unless the context indicates otherwise, it is specifically intended that the various features of the invention described herein can be used in any combination. Moreover, the present invention also contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted. To illustrate, if the specification states that a composition comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, can be omitted and disclaimed singularly or in any combination.
The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if the range 10-15 is disclosed, then 11, 12, 13, and 14 are also disclosed. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the disclosure and does not pose a limitation on the scope of the disclosure unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.
As used in the description of the invention and the appended claims, the singular forms “a,” “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.
Also as used herein, “and/or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).
The term “about,” as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of the specified value as well as the specified value. For example, “about X” where X is the measurable value, is meant to include X as well as variations of ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of X. A range provided herein for a measurable value may include any other range and/or individual value therein.
As used herein, phrases such as “between X and Y” and “between about X and Y” should be interpreted to include X and Y. As used herein, phrases such as “between about X and Y” mean “between about X and about Y” and phrases such as “from about X to Y” mean “from about X to about Y.”
The term “comprise,” “comprises” and “comprising” as used herein, specify the presence of the stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.
As used herein, the transitional phrase “consisting essentially of” means that the scope of a claim is to be interpreted to encompass the specified materials or steps recited in the claim and those that do not materially affect the basic and novel characteristic(s) of the claimed invention. Thus, the term “consisting essentially of” when used in a claim of this invention is not intended to be interpreted to be equivalent to “comprising.”
As used herein, the terms “increase,” “increasing,” “increased,” “enhance,” “enhanced,” “enhancing,” and “enhancement” (and grammatical variations thereof) describe an elevation of at least about 5%, 10%, 15%, 20%, 25%, 50%, 75%, 100%, 150%, 200%, 300%, 400%, 500% or more as compared to a control.
As used herein, the terms “reduce,” “reduced,” “reducing,” “reduction,” “diminish,” and “decrease” (and grammatical variations thereof), describe, for example, a decrease of at least about 5%, 10%, 15%, 20%, 25%, 35%, 50%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% as compared to a control. In particular embodiments, the reduction can result in no or essentially no (i.e., an insignificant amount, e.g., less than about 10% or even 5%) detectable activity or amount.
As used herein, “a plant” can be, but is not limited to, any monocotyledonous and dicotyledonous plant, and any annual and perennial dicotyledonous and monocotyledonous plant. Example plants include, but are not limited to, those of the genera Glycine, Vitis, Asparagus, Populus, Pennisetum, Lolium, Oryza, Zea, Avena, Hordeum, Secale, Triticum, Helianthus, Gossypium, Medicago, Pisum, Acer, Actinidia, Abelmoschus, Agropyron, Allium, Amaranthus, Apium, Arachis, Asparagus, Beta, Brassica, Camellia, Canna, Capsicum, Carex, Carica papaya, Carya, Castanea, Cinnamomum, Citrullus, Citrus, Cocos, Coffea, Colocasia, Cola, Coriandrum, Corylus, Crataegus, Crocus, Cucurbita, Cucumis, Cynara, Daucus, Desmodium, Dimocarpus, Dioscorea, Diospyros, Echinochloa, Elaeis, Eleusine, Eriobotrya, Eugenia, Fagopyrum, Fagus, Ficus, Fortunella, Fragaria, Ginkgo, Hemerocallis, Hibiscus, Ipomoea, Juglans, Lactuca, Lathyrus, Lens, Linum, Litchi, Lotus, Lupinus, Luzula, Malus, Malpighia, Mammea, Mangifera, Manihot, Manilkara, Medicago, Melilotus, Mentha, Miscanthus, Musa, Nicotiana, Olea, Opuntia, Ornithopus, Panicum, Passiflora, Persea, Phaseolus, Pinus, Pistacia, Pisum, Poa, Prosopis, Prunus, Quercus, Raphanus, Rheum, Ribe, Rubus, Sambucus, Secale, Sesamum, Sinapis, Solanum, Sorghum, Spinacia, Tamarindus, Theobroma, Trifolium, Tropaeolum, Vaccinium, Vigna, Vitis, Zizania or Ziziphus Sorghum, Saccharum and Lycopersicum, or the class Liliatae. In some embodiments, a plant or part thereof is from the genera Glycine, Vitis, Asparagus, Populus, Pennisetum, Lolium, Oryza, Zea, Avena, Hordeum, Secale, Triticum, Sorghum, Saccharum and Lycopersicum, or the class Liliaceae.
As used herein, a “part thereof” includes, but is not limited to, plant reproductive tissues (e.g., petals, sepals, stamens, pistils, receptacles, anthers, pollen, flowers, fruits, flower buds, ovules, seeds, embryos, nuts, kernels, bulbs, ears, cobs and husks); plant vegetative tissues (e.g., petioles, stems, roots, root hairs, root tips, pith, coleoptiles, stalks, shoots, branches, bark, apical meristem, axillary bud, cotyledon, hypocotyls, and leaves); plant vascular tissues (e.g., phloem and xylem); specialized plant cells such as epidermal cells, parenchyma cells, collenchyma cells, sclerenchyma cells, stomata, guard cells, cuticle, mesophyll cells; callus tissue; and cuttings. It also includes plant cells, including plant cells that are intact in plants and/or parts of plants, plant protoplasts, plant tissues, plant organs, plant cell tissue cultures, plant calli, plant clumps, and the like.
A “growth media” or “growing media”, as used herein includes, but is not limited to soil, synthetic growth media and/or aqueous solution in which the plant is planted (e.g., a hydroponic solution). A growing media may be treated/contacted with a composition comprising MLGs before and/or after sowing seedlings or seeds including spraying or irrigating the growth medium with a composition comprising MLGs. For example, a composition comprising MLGs formulated into a liquid or solid formulation, such as a granule, may be applied to soil around seedlings prior to, during and/or after sowing the seedlings. In some aspects, both a growth medium and the plant or plant part planted or sowed in the same may be treated/contacted with a composition comprising MLGs.
As used herein, “growth characteristic” refers to any plant trait associated with growth, for example, biomass, root mass, yield, inflorescence size/weight, fruit yield, fruit quality, fruit size, seed production, foliar tissue weight, nodulation number, nodulation mass, nodulation activity, number of seed heads, number of tillers, number of flowers, number of tubers, tuber mass, bulb mass, number of seeds, total seed mass, rate of leaf emergence, rate of tiller emergence, rate of seedling emergence or any combination thereof. Thus, in some aspects, an increased growth characteristic may include, but is not limited to, increased fruit production, increased inflorescence production, increased fruit quality, increased root mass and/or biomass as compared to a control plant and/or part thereof and/or growth media where a composition comprising MLGs has not been applied.
“Fruit quality” refers to the typical fruit quality characteristics such as appearance: fruit size, diameter (e.g., inches, centimeters) and/or weight (g). Other fruit quality characteristics can include, but are not limited to, color (e.g., lycopene content in tomatoes), taste (sweetness, sourness, bitterness, etc.) and/or nutrient and/or nutritional content (e.g., sugar content (Brix value), proteins, lipids, vitamins, and/or minerals, etc.).
As used herein, “nutrient use efficiency” refers to a plant’s ability to utilize available nutrients. In some embodiments, “nutrient use efficiency may be defined in terms of total nutrient uptake (nutrient concentration in plant tissue × total biomass) and/or yield per unit of nutrient applied.
The term “abiotic stress” as used herein refers to outside, nonliving, factors which can cause harmful effects to plants. Thus, as used herein, abiotic stress includes, but is not limited to, cold temperature that results in freezing, chilling, heat or high temperatures, drought, high light intensity, low light intensity, salinity, flooding (excess water/water-logging), ozone, and/or combinations thereof. Parameters for the abiotic stress factors are species specific and even variety specific and therefore vary widely according to the species/variety exposed to the abiotic stress. Thus, while one species may be severely impacted by a high temperature of 23° C., another species may not be impacted until at least 30° C., and the like. Temperatures above 30° C. result in dramatic reductions in the yield of most plants. This is due to reductions in photosynthesis that begin at approximately 20-25° C., and the increased carbohydrate demands of crops growing at higher temperatures. The critical temperatures are not absolute, but vary depending upon such factors as the acclimatization of plants to prevailing environmental conditions. In addition, because most plants are exposed to multiple abiotic stresses at one time, the interaction between the stresses affects the response of the plant. Thus, the particular parameters for high/low temperature, light intensity, drought and the like, which impact plant productivity will vary with species, variety, degree of acclimatization and the exposure to a combination of environmental conditions.
The inventors of the present invention have discovered that treating a plant and/or part thereof, and/or a growth medium for growing a plant, with a composition comprising an effective amount/rate of MLGs can increase a growth characteristic, nutrient use efficiency, and/or abiotic and/or biotic stress tolerance/resistance of the plant and/or part thereof.
In some embodiments, a composition may comprise a single MGL or a mixture of MLGs with a degree of polymerization (DP) in a range from about DP3 to about DP >100. Thus, in some embodiments, a composition of the invention may comprise an MLG of DP3 (also referred to herein as “MLG43”), and/or DP4, and/or DP5, and/or DP6, and/or DP7, and/or DP8, and/or DP9, and/or DP10, and/or DP11, and/or DP12, and/or DP13, and/or DP14, and/or DP15, and/or DP16, and/or DP17, and/or DP18, and/or DP20, and/or DP21, and/or DP22, and/or DP23, and/or DP24, and/or DP25, and/or DP26, and/or DP27, and/or DP28, and/or DP29, and/or DP30, and/or DP40, and/or DP50, and/or DP60, and/or DP70, and/or DP80, and/or DP90, and/or DP100, and/or DP>100 and any range or value therein. In some embodiments, a composition may comprise a mixture of MLGs of DP from about DP3 to about DP6, from about DP3 to about DP7, from about DP3 to about DP8, from about DP3 to about DP9, from about DP3 to about DP10, from about DP3 to about DP15, about DP3 to about DP20, about DP3 to about DP30, about DP3 to about DP40, about DP3 to about DP50, about DP3 to about DP60, about DP3 to about DP70, about DP3 to about DP80, about DP3 to about DP90, about DP3 to about DP100, from about DP5 to about DP8, from about DP5 to about DP9, from about DP5 to about DP10, from about DP5 to about DP15, about DP5 to about DP20, about DP5 to about DP50, about DP5 to about DP40, about DP5 to about DP50, about DP5 to about DP60, about DP5 to about DP70, about DP5 to about DP80, about DP5 to about DP90, about DP5 to about DP100, about DP10 to about DP15, about DP10 to about DP20, about DP10 to about DP30, about DP10 to about DP40, about DP10 to about DP50, about DP10 to about DP60, about DP10 to about DP70, about DP10 to about DP80, about DP10 to about DP90, about DP10 to about DP100, about DP20 to about DP30, about DP20 to about DP40, about DP120 to about DP50, about DP20 to about DP60, about DP20 to about DP70, about DP20 to about DP80, about DP20 to about DP90, about DP20 to about DP100, DP40 to about DP50, about DP40 to about DP60, about DP40 to about DP70, about DP40 to about DP80, about DP40 to about DP90, about DP40 to about DP100, DP50 to about DP70, about DP50 to about DP80, about DP50 to about DP90, about DP50 to about DP100, and any range or value therein.
In some embodiments, a composition may comprise an amount of MLGs (comprising a single degree of polymerization or a mixture of degree of polymerization) in a range from about 0.1 mg per liter (L) to about 100 g per liter of the composition. Thus, in some embodiments, a composition of the invention may comprise MLGs in a range from about 0.1 mg/L to about 1 mg/L, about 0.1 mg/L to about 10 mg/L, about 0.1 mg/L to about 100 mg/L, about 0.1 mg/L to about 1 g/L, about 0.1 mg/L to about 10 g/L, about 0.1 mg/L to about 100 g/L, about 0.1 g/L to about 1 g/L, about 0.1 g/L to about 5 g/L, about 0.1 g/L to about 10 g/L, about 0.1 g/L to about 15 g/L, about 0.1 g/L to about 20 g/L, about 0.1 g/L to about 30 g/L, about 0.1 g/L to about 40 g/L, about 0.1 g/L to about 100 g/L, about 0.5 g/L to about 1 g/L, about 0.5 g/L to about 5 g/L, about 0.5 g/L to about 10 g/L, about 0.5 g/L to about 20 g/L, about 0.5 g/L to about 30 g/L, about 0.5 g/L to about 40 g/L, about 0.5 g/L to about 50 g/L, about 0.5 g/L to about 100 g/, about 1 g/L to about 5 g/L, about 1 g/L to about 10 g/L, about 1 g/L to about 15 g/L, about 1 g/L to about 20 g/L, about 1 g/L to about 30 g/L, about 1 g/L to about 40 g/L, about 1 g/L to about 50 g/L, about 1 g/L to about 100 g/L, about 5 g/L to about 10 g/L, about 5 g/L to about 15 g/L, about 5 g/L to about 20 g/L, about 5 g/L to about 30 g/L, about 5 g/L to about 40 g/L, 1, about 5 g/L to about 50 g/L, about 5 g/L to about 100 g/L, about 10 g/L to about 15 g/L, about 10 g/L to about 20 g/L, about 10 g/L to about 30 g/L, about 10 g/L to about 40 g/L, about 10 g/L to about 50 g/L, about 10 g/L to about 100 g/L, about 15 g/L to about 20 g/L, about 15 g/L to about 30 g/L, about 15 g/L to about 40 g/L, about 15 g/L to about 50 g/L, about 15 g/L to about 100 g/L, about 20 g/L to about 30 g/L, or about 20 g/L to about 40 g/L, about 20 g/L to about 50 g/L, about 20 g/L to about 100 g/L, about 30 g/L to about 40 g/L, about 30 g/L to about 50 g/L,, about 30 g/L to about 100 g/L, about 40 g/L to about 50 g/L, or about 40 g/L to about 100 g/L of the composition, or any value or range therein.
In some embodiments, an effective amount of MLGs is an amount sufficient to increase a growth characteristic of a plant and/or part thereof, increase nutrient use efficiency in a plant and/or part thereof, and/or increase abiotic stress and/or biotic stress tolerance/resistance of a plant and/or part thereof. In some embodiments, an effective amount of MLGs in a composition may be from about 0.1 mg per liter to about 100 g per liter of the composition. In some embodiments, an effective amount of MLGs in a composition may be from about 0.1 mg/L to about 1 mg/L, about 0.1 mg/L to about 10 mg/L, about 0.1 mg/L to about 100 mg/L, about 0.1 mg/L to about 1 g/L, about 0.1 mg/L to about 10 g/L, about 0.1 mg/L to about 100 g/L, about 0.1 g/L to about 1 g/L, about 0.1 g/L to about 5 g/L, about 0.1 g/L to about 10 g/L, about 0.1 g/L to about 15 g/L, about 0.1 g/L to about 20 g/L, about 0.1 g/L to about 30 g/L, about 0.1 g/L to about 40 g/L, about 0.1 g/L to about 100 g/L, about 0.5 g/L to about 1 g/L, about 0.5 g/L to about 5 g/L, about 0.5 g/L to about 10 g/L, about 0.5 g/L to about 20 g/L, about 0.5 g/L to about 30 g/L, about 0.5 g/L to about 40 g/L, about 0.5 g/L to about 50 g/L, about 0.5 g/L to about 100 g/, about 1 g/L to about 5 g/L, about 1 g/L to about 10 g/L, about 1 g/L to about 15 g/L, about 1 g/L to about 20 g/L, about 1 g/L to about 30 g/L, about 1 g/L to about 40 g/L, about 1 g/L to about 50 g/L, about 1 g/L to about 100 g/L, about 5 g/L to about 10 g/L, about 5 g/L to about 15 g/L, about 5 g/L to about 20 g/L, about 5 g/L to about 30 g/L, about 5 g/L to about 40 g/L, 1, about 5 g/L to about 50 g/L, about 5 g/L to about 100 g/L, about 10 g/L to about 15 g/L, about 10 g/L to about 20 g/L, about 10 g/L to about 30 g/L, about 10 g/L to about 40 g/L, about 10 g/L to about 50 g/L, about 10 g/L to about 100 g/L, about 15 g/L to about 20 g/L, about 15 g/L to about 30 g/L, about 15 g/L to about 40 g/L, about 15 g/L to about 50 g/L, about 15 g/L to about 100 g/L, about 20 g/L to about 30 g/L, or about 20 g/L to about 40 g/L, about 20 g/L to about 50 g/L, about 20 g/L to about 100 g/L, about 30 g/L to about 40 g/L, about 30 g/L to about 50 g/L,, about 30 g/L to about 100 g/L, about 40 g/L to about 50 g/L or about 40 g/L to about 100 g/L of the composition, or any value or range therein.
In some embodiments, a composition comprising MLGs may further comprise a peptide, a protein, a sugar and/or a carbohydrate. In some embodiments, a MLGs composition may comprise peptides and/or proteins in an amount from about 0.1% to about 10% w/w of the extract. In some embodiments, a composition comprising MLGs comprise peptides and/or proteins in an amount from about 0.1% to about 1%, about 0.1% to about 3%, about 0.1% to about 5%, about 0.1% to about 7%, about 0.5% to about 1%, about 0.5% to about 3%, about 0.5% to about 5%, about 0.5% to about 7%, about 0.5 to about 10%, about 1% to about 3%, about 1% to about 5%, about 1% to about 7%, about 1% to about 10%, about 3% to about 5%, about 3% to about 7%, about 3% to about 10%, about 5% to about 7%, about 5% to about 10%, or about 7% to about 10%, or any range or value therein of the composition. Thus, in some embodiments, a composition comprising MLGs may comprise peptides and/or proteins in an amount of about 0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10% w/w or any range or value therein of the composition.
In some embodiments, a composition comprising MLGs may comprise others sugars and/or carbohydrates in an amount from about 1% to about 35% w/w of the extract. In some embodiments, a composition comprising MLGs may comprise others sugars and/or carbohydrates in an amount from about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 25% to about 30%, about 25% to about 35%, or about 30% to about 35% w/w of the extract, or any value or range therein. Thus, in some embodiments, a composition comprising MLGs may comprise additional sugars and/or carbohydrates in an amount of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35% w/w of the extract or any range or value therein.
In some embodiments, sugars and/or carbohydrates that may be comprised in a composition comprising MLGs can include, but is not limited to glucose, mannose, galactose, arabinose, xylose other glucan oligosaccharides, glucose-derived low branched polysaccharides, glycogen, mannan oligosaccharides, mannose-derived low branched polysaccharides, galactans, galactomannans, arabinans and/or xylans.
In some embodiments, a composition comprising MLGs may further comprise additional components including, but not limited to, a surfactant, a humectant, an adjuvant, an antioxidant, a preservative, a plant macronutrient, a plant micronutrient, a plant growth regulator, a pesticide, a fungicide, an antiviral, an anti-bacterial, an herbicide, or any combination thereof.
Example surfactants can include, but are not limited to, alkali metal, alkaline earth metal and ammonium salts of ligno-sulfonic acid, naphthalenesulfonic acid, phenolsulfonic acid, dibutylnaphthalenesulfonic acid, alkylarylsulfonates, sodium dodecylsulfate, alkyl sulfates, alkylsulfonates, fatty alcohol sulfates, fatty acids and sulfated fatty alcohol glycol ethers, of sulfonated condensates naphthalene and naphthalene derivatives with formaldehyde, condensates of naphthalene or of naphthalenesulfonic acid with phenol and formaldehyde, polyoxyethylene octyl-phenyl ether, ethoxylated isooctylphenol, octylphenol, nonylphenol, alkylphenyl poly-glycol ethers, tributylphenyl polyglycol ether, tristearylphenyl polyglycol ether, alkylaryl polyether alcohols, alcohol and fatty alcohol/ethylene oxide condensates, ethoxylated castor oil, polyoxyethylene alkyl ethers, ethoxylated polyoxypropylene, lauryl alcohol, polyglycol ether acetal, sorbitol esters, lignin-sulfite waste liquors and/or methylcellulose.
In some embodiments, a surfactant may be present in a composition comprising MLGs in an amount from about 10% to about 40% w/w of the composition. In some embodiments, a surfactant may be present in a composition comprising MLGs in an amount from about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 15% to about 40%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 20% to about 40%, about 25% to about 30%, about 25% to about 35%, about 25% to about 40%, about 30% to about 35%, about 30% to about 40%, about 35% to about 40% w/w of the composition or any range or value therein). Thus, in some embodiments, the surfactant may be present in a composition comprising MLGs in an amount of about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40% w/w of the composition or any range or value therein.
Example humectants can include, but are not limited to, glycerol, sorbitol, xylitol, maltitol, glyceryl triacetate, sodium lactate, urea formaldehyde, propylene glycol, ethylene glycol and/or fatty acids.
An example antioxidant can include, but is not limited to, ascorbic acid, tocopherols, propyl gallate, tertiary butylhydroquinone, butylated hydroxyanisole, and/or butylated hydroxytoluene.
An example preservative can include, but is not limited to, sorbic acid, sodium sorbate, sorbates, benzoic acid, sodium benzoate, benzoates, hydroxybenzoate and derivatives, sulfur dioxide and sulphites, nitrite, nitrate, lactic acid, propionic acid and sodium propionate, tocopherol, plant extract, hops, salt, sugar, vinegar, alcohol (e.g. methanol and ethanol), diatomaceous earth and castor oil, citric acid, ascorbic acid, sodium ascorbate, phenol derivatives (butylated hydroxytoluene, butylated hydroxyanisole, BHA, BHT, TBHQ and propyl gallate), gallic acid, sodium gallate, sulfur dioxide, sulphites, tocopherols, and/or methylchloroisothiazolinone, 1,2-Benzisothiazolin-3-one (BIT), Hexahydro-1,3,5-tris-hydroxyethyl -s-triazine (HTHT), 5-chloro-2-methyl-2H-isothiazol-3-one (CMIT), 2-methyl-2H-isothiazol-3-one (MIT), Zinc pyrithione (ZPT), 2-Bromo-2-nitropropane-1,3-diol (Bronopol), Formaldehyde, 1,3-Dimethylol-5,5-dimethylhydantoin (DMDMH), 2,2-Dibromo-3-nitrilopropionamide (DBNPA), and/or Poly (hexamethylene biguanide) hydrochloride (PHMB).
In some embodiments, a preservative may be present in a composition comprising MLGs in an amount ranging from about 0.001% to about 5% w/w or any range or value therein. In some embodiments, a composition may comprise a preservative in an amount ranging from about 0.001% to about 0.1%, about 0.001% to about 0.5%, about 0.001% to about 1%, about 0.001% to about 2%, about 0.001% to about 3%, about 0.001% to about 4%, about 0.01% to about 0.1%, about 0.01% to about 0.5%, about 0.01% to about 1%, about 0.01% to about 2%, about 0.01% to about 3%, about 0.01% to about 4%, about 0.01% to about 5%, about 0.05% to about 0.1%, about 0.05% to about 0.5%, about 0.05% to about 1%, about 0.05% to about 2%, about 0.05% to about 3%, about 0.05% to about 4%, about 0.05% to about 5%, about 0.1% to about 0.5%, about 0.1% to about 1%, about 0.1% to about 2%, about 0.1% to about 3%, about 0.1% to about 4%, about 0.1% to about 5%, about 0.5% to about 1%, about 0.5% to about 2%, about 0.5% to about 3%, about 0.5% to about 4%, about 0.5% to about 5%, about 1% to about 2%, about 1% to about 3%, about 1% to about 4%, about 1% to about 5%, about 2% to about 3%, about 2% to about 4%, about 4% to about 5%, about 3% to about 4%, about 3% to about 5%, about 4% to about 5% w/w of the composition, or any range or value therein. Thus, in some embodiments, the preservative may be present in a composition comprising MLGs in an amount of about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5% w/w of the composition or any range or value therein.
Example plant macronutrients include, but are not limited to, nitrogen, potassium, calcium, magnesium, phosphorus, and/or sulfur.
Example plant micronutrients can include, but are not limited to, iron, manganese, boron, molybdenum, copper, zinc, chlorine, and/or cobalt.
Example plant growth regulators include, but are not limited to, auxin (including but not limited to naphthalene acetic acid (NAA) and/or indole-3-butyric acid (IBA) and/or indole-3-acetic acid (IAA, 3-IAA)), cytokinin, abscisic acid, gibberellin, ethylene, salicylic acid, jasmonic acid, brassinosteriod (e.g., brassinolide), or any combination thereof.
Example pesticides include, but are not limited to, malathion, parathion, methyl parathion, chlorpyrifos, diazinon, dichlorvos, phosmet, fenitrothion, tetrachlorvinphos, azamethiphos, fenvalerate, cyfluthrin, lambda-cyhalothrin, zeta-cypermethrin, permethrin, piperonyl butoxide, imidacloprid, acetamiprid, clothianidin, nitenpyram, nithiazine, thiacloprid, thiamethoxam, ryanodol, 9,21-didehydroryanodol, chlorantraniliprole, flubendiamide, and/or cyantraniliprole.
Example fungicides include, but are not limited to, prothioconazole, trifloxystrobin, azoxystrobin, propiconazole, and/or pyraclostrobin.
Example anti-bacterials (bactericides) include, but are not limited to, methylisothiazolinone, chloromethylisothiazolinone, benzisothiazolinone, octylisothiazolinone, dichlorooctylisothiazolinone, and/or butylbenzisothiazolinone
Example herbicides can include, but are not limited to, glyphosate, 2,4-dichlorophenoxyacetic acid, atrazine, S-metolachlor, and/or 3,6-dichloro-2-methoxybenzoic acid.
In some embodiments, a composition comprising MLGs may further comprise an antifoaming agent. Any antifoaming agent for use with agricultural and/or food products may be used. Example antifoaming agents include, but are not limited to, long chain unsaturated fatty acids including, but not limited to C12 to C14, C18:1 and C18:2 unsaturated fatty acids, and/or synthetic polysiloxanes (silicones) including, but not limited to, polydimethylsiloxane, and/or hydrophobic silica. In some embodiments, a composition comprising MLGs may comprise an amount of antifoaming agent in a range from about 0.0001% to about 0.05% w/w of the composition or any range or value therein. Thus, in some embodiments, the antifoaming agent may be present in the composition in an amount of about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, or 0.05% w/w of the composition or any range or value therein.
In some embodiments, a composition comprising MLGs may further comprise a biocide. Any biocide for use with agricultural and/or food products may be used. When included in a composition comprising MLGs, a biocide may be present at about 0.1 g L-1 to about 20 g L-1 of the composition, or any range or value therein. Thus, in some embodiments, the biocide may be present in the composition in an amount of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 g per liter of the composition, or any range or value therein.
A composition comprising MLGs may be applied to plant and/or part thereof. In some embodiments, a composition comprising MLGs may be atomized and sprayed on a plant and/or part thereof. A smearing treatment may also be used, for example, where a wettable powder, an emulsion or a flowable agent of a composition comprising MLGs may be applied to a plant and/or plant part thereof with or without water added. In another embodiments, an immersing treatment may be used in which a plant and/or part thereof are immersed in a composition comprising MLGs. In some embodiments, a composition comprising MLGs may be used in a film coating treatment and/or a pellet coating treatment for a plant and/or part hereof such as seeds and/or bulbs.
A composition comprising MLGs may be applied as a soil treatment in the form of a solid or a liquid. Thus, in some embodiments, the composition may be applied as a spray onto soil, soil incorporation, and/or perfusion of a chemical liquid into the soil (irrigation of chemical liquid, soil injection, and dripping of chemical liquid). Methods for applying a composition comprising MLGs during a soil treatment can include, but are not limited to, planting hole, furrow, around a planting hole, around a furrow, entire surface of cultivation lands, the parts between the soil and the plant, area between roots, area beneath the trunk, main furrow, growing box, seedling growing tray and seedbed. A soil treatment may be applied before seeding, at the time of seeding, immediately after seeding, germination and/or growing period after planting. Alternatively, an irrigation liquid may be mixed with a composition comprising MLGs in advance and, for example, used for treatment by an appropriate irrigating method including the irrigation method mentioned above and any other methods such as methods and compositions to improve plant health and protection.
A composition comprising MLGs may be applied to a plant and/or a plant part thereof, and/or to a medium in which a plant is growing for increasing, for example, a growth characteristic, nutrient use efficiency, disease tolerance (biotic stress, e.g., tolerance to fungal, bacterial, and/or viral diseases) and/or for increasing abiotic stress tolerance. Thus, in some embodiments, the present invention provides a method for increasing a growth characteristic of a plant and/or part thereof, the method comprising applying a composition comprising an effective amount of MLGs to a plant and/or part thereof and/or growth media, thereby increasing the growth characteristic of the plant and/or part thereof as compared to a control plant and/or part thereof (e.g., a plant and/or part thereof to which the composition of the invention has not been applied). In some embodiments, the method comprises applying a composition comprising MLGs to a plant and/or part thereof, and/or a growth media in which the plant and/or part thereof is present at least one time (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and/or 12 or more times; e.g., 1 to about 2 times, 1 to about 3 times, 1 to about 4 times, 1 to about 5, times, 1 to about 6 times, 1 to about 7 times, 1 to about 8 times, 1 to about 9 times, 1 to about 10 times, 1 to about 11 times, or 1 to about 12 times, or any range or value therein). In some embodiments, the method comprises applying the composition at least two times (e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times; e.g., about 2 to about 3 times, about 2 to about 4 times, about 2 to about 5, times, about 2 to about 6 times, about 2 to about 7 times, about 2 to about 8 times, about 2 to about 9 times, about 2 to about 10 times, about 2 to about 11 times, or about 2 to about 12 times, or any range or value therein).
In some embodiments, the present invention provides a method for increasing disease tolerance of a plant and/or part thereof, the method comprising applying a composition comprising an effective amount of MLGs to a plant and/or a plant part thereof, and/or to a growth media, thereby increasing the disease tolerance of the plant and/or part thereof as compared to a control plant and/or part thereof (e.g., a plant and/or part thereof to which a composition comprising MLGs has not been applied). In some embodiments, the method comprises applying the composition at least one time (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times). In some embodiments, the method comprises applying the composition at least two times (e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times).
In some embodiments, when a composition comprising MLGs is applied at least two times to a plant and/or part thereof, and/or growth media, the range of time between application may vary. Thus, for example, a subsequent application of a composition comprising MLGs may be any time from about 1 day to about six month after the prior application. Thus, for example, a subsequent application may be applied about 1, 2, 3, 4, 5, 6 days or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 weeks after previous application, or any range or value therein.
In some embodiments, a composition comprising MLGs applied to a plant and/or part thereof, and/or to a growth media, may increase disease tolerance or resistance to a viral pathogen including, but not limited to, a virus from the virus family of Caulimoviridae, Potyviridae, Sequiviridae, Rheoviridae, Capillovirus, Geminiviridae, Bromoviridae, Closteroviridae, Comoviridae Tombusviridae, Rhabdoviridae, Bunyaviridae, Partitiviridae, Carlavirus, Enamovirus, Furovirus, Hordeivirus, Idaeovirus, Luteovirus, Marafivirus, Potexvirus, Sobemovirus, Tenuivirus, Tobamovirus, Tobravirus, Trichovirus, Tymovirus and/or Umbravirus.
In some embodiments, a composition comprising MLGs applied to a plant and/or part thereof, and/or to a growth media, may increase resistance to a virus, including but not limited to, turnip mosaic virus, papaya ring spot virus, bud blight virus, bean pod mottle virus, lettuce mosaic virus, maize mosaic virus, cauliflower mosaic virus, tobacco mosaic virus, soybean mosaic virus, African cassava mosaic virus, tomato mosaic virus, pepino mosaic virus, zucchini yellow mosaic virus, plum pox virus, tomato bushy stunt virus, tomato spot wilt virus, tomato yellow leaf curl virus, rice ragged stunt virus, rice tungro bacilliform, virus, rice tungro spherical virus, rice yellow mottle virus, cucumber mosaic virus, brome mosaic virus, wheat yellow mosaic virus, barley yellow dwarf virus, sugarcane mosaic virus, beet yellows virus, lettuce yellows virus, maize dwarf mosaic virus, maize streak virus, peanut stunt virus, Citrus tristeza virus, potato leafroll virus, potato virus X, potato virus Y, sweet potato feathery mottle potyvirus, Melon necrotic spot virus, maize white line mosaic virus, maize chlorotic mottle virus, banana bunchy top virus, cacao swollen shoot virus, tomato leaf curl New Dehli virus, banana streak virus, and/or sweet potato sunken vein closterovirus.
In some embodiments, a composition comprising MLGs applied to a plant and/or part thereof, and/or to a growth media, may increase resistance to a fungal pathogen including, but not limited to, a fungal/oomycete pathogen from the family of Physodermataceae, Synchytriaceae, Olpidiaceae, Choanephoraceae, Gilbertellaceae, Mucoraceae, Dipodascaceae, Eremotheciaceae, Taphrinaceae, Botryosphaeriaceae, Capnodiaceae, Phaeosphaeriaceae, Leptosphaeriaceae, Cucurbitariaceae, Didymellaceae Davidiellaceae, Mycosphaerellaceae, Schizothyriaceae, Dothideaceae, Dothioraceae, Lahmiaceae, Elsinoaceae, Lophiostomataceae, Pleosporaceae, Venturiaceae, Trichochomaceae, Erysiphaceae, Cyttariaceae, Hemiphacidiaceae, Hyaloscyphaceae, Phacidiaceae, Sclerotiniaceae, Ascodichaenaceae, Mediolariaceae, Rhytismataceae, Meliolaceae, Caloscyphaceae, Sarcosomataceae, Cryphonectriaceae, Diaporthaceae, Gnomoniaceae, Valsaceae, Glomerellaceae, Plectosphaerellaceae, Bionectriaceae, Clavicipitaceae, Hypocreaceae, Nectriaceae, Magnaporthaceae, Pyriculariaceae, Ceratocystideae, Ophiostomataceae, Phyllachoraceae, Chaetomiaceae, Amphisphaeriaceae, Diatrypaceae, Xylariaceae, Psathyrellaceae, Marasmiaceae, Mycenaceae, Schizophyllaceae, Typhulaceae, Thelephoraceae, Atheliaceae, Atheliaceae, Stereaceae, Echinodontiaceae, Corticiaceae, Ganodermataceae, Hymenochaetaceae, Cystofilobasidiaceae, Helicobasidiaceae, Helicobasidiaceae, Melampsoraceae, Phakopsoraceae, Pucciniaceae, Tilletiaceae, Entylomataceae, Ustilaginaceae, Leptolegniaceae and/or Peronosporaceae.
In some embodiments, a composition comprising MLGs applied to a plant and/or part thereof, and/or to a growth media, may increase resistance to a fungal pathogen including, but not limited to, Physoderma alfalfa, Physoderma maydis, Synchytrium endobioticum, Olpidium brassicae, Choanephora cucurbitarum, Mucor circinelloides, Rhizopus stolonifera, Geotrichum candidum, Taphrina caerulescens, Taphrina deformans, Taphrina populina, Botryosphaeria dothidea, Diplodia mutila, Dothiorella sarmentorum, Macrophomina phaseolina, Phyllosticta ampelicida, Phyllosticta citricarpa, Stenocarpella maydis, Cladosporium allii-cepae, Cladosporium cladosporioides, Acrodontium simplex, Cercospora spp., Cercospora apii, Cercospora beticola, Cercospora brassicicola, Cercospora kikuchii, Corynespora cassiicola, Cercospora zeae-maydis, Cercospora zeina, Dothistroma septosporum, Lecanosticta acicula, Mycocentrospora acerina, Passaloraspp., Pseudocercospora fijiensis, Aureobasidiumspp., Ophiosphaerella herpotricha, Parastagonospora nodorum, Diplodia tumefaciens, Alternaria alternate, Bipolaris maydis, Bipolaris oryzae, Bipolaris sacchari, Bipolaris victoriae, Curvulariaspp., Leptosphaerulina trifolii, Venturia inaequalis, Aspergillusspp., Aspergillus flavus, Blumeria graminis, Erysiphespp., Podosphaera leucotricha, Botrytis cinerea, Moniliniaspp., Monilinia fructicola, Sclerotinia sclerotiorum, Amphilogia gyrosa, Cryphonectria parasitica, Diaporthe citri, Diaporthe helianthi, Diaporthe phaseolorum, Cytospora leucostoma, Colletotrichumspp., Colletotrichum coccodes, Colletotrichum gloeosporioides, Colletotrichum graminicola, Plectosphaerella cucumerina, Verticillium albo-atrum, Verticillium dahlia, Claviceps purpurea, Epichloe typhina, Trichoderma viride, Fusariumspp., Fusarium oxysporum, Fusarium solani, Fusarium graminearum, Nectria cinnabarina, Neonectriaspp., Gaeumannomyces graminis, Pyricularia grisea, Pyricularia oryzae, Ceratocystisspp., Thielaviopsis basicola, Ophiostoma ulmi, Phyllachora graminis, Cronartiumspp., Uromyces graminicola, Tranzscheliaspp., Tilletiaspp., Ustilagospp., Ustilago maydis, Peronosporaspp., Hyalperonosporaspp., Albugospp., Phytophthoraspp., Pythiumspp., Aphanomycesspp., Magnaporthe oryzae, Puccinia, Blumeria graminis, Exserohilum turcicum, Mycosphaerella graminicola, Melampsora lini, Phakopsora pachyrhizi, and/or Rhizoctonia solani.
In some embodiments, a composition comprising MLGs applied to a plant and/or part thereof, and/or to a growth media, may increase resistance to a bacterial pathogen including, but not limited to, a bacterial pathogen from the family of Enterobacteriaceae, Pseudomonadaceae, Rhizobiaceae, Microbacteriaceae, Xanthomonadaceae, Rhizobiaceae, Corynebacteriaceae, Acetobacteraceae, Comamonadaceae, Bacillaceae, Burkholderiaceae, Micrococcaceae, Ralstoniaceae, Xanthomonadaceae, Spiroplasmataceae, Sphingomonadaceae, Acholeplasmataceae, Corynebacteriaceae, and/or Streptomycetaceae. In some embodiments, a composition of the invention applied to a plant and/or part thereof, and/or growth media, may increase resistance to a bacterial pathogen including, but not limited to, a bacterial pathogen from the genus of Erwinia spp., Dickeya spp., Pseudomonas spp., Xanthomonas spp., Agrobacterium spp., Rhizobium spp., Corynebacterium spp., Streptomyces spp., Pantoea spp., Serratia spp., Acetobacter spp., Acidovorax spp., Arthrobacter spp., Bacillus spp., Brenneria spp., Burkholderia spp., Clavibacter spp., Pectobacterium spp., Pantoea spp., Ralstonia spp., Xylella spp., Spiroplasma spp., Phytoplasma spp., and/or Sphingomonas spp.
In some embodiments, a composition comprising MLGs applied to a plant and/or part thereof, and/or to a growth medium, may increase resistance to a bacterial pathogen including, but not limited to, Erwinia amylovora, E.acarotovoravar. chrysanthemi, D. dadanti, Pseudomonas tabaci, P. angulate, P. phaseolicola, P. lachrymans, P. pisi, P. fluorescens, P. glycinea, P. vesicatoria, P. savastanoi, P. syringae, P. solanacearum, Xanthamonas phaseoli, X. malvacearum, X. oryzae, Xtranslucens, X. pruni, X. campestris, X. vasuclarum, Acidovorax avenae, Agrobacterium tumefaciens, A. rubi (=Rhizobium rubi), A. rhizogenes (=Rhizobium rhizogenes) and A. vitis (=Rhizobium vitis), Bacillus pumilus., Brenneria alni (= Erwinia alni), Clavibacter michiganensis, Pectobacterium carotovorum, Pantoea agglomerans, Ralstonia solanacearum, Corynebacterium insidiosum, C. sepedonicum, C. fascians, C. flacumfaciens, C. michiganense, Streptomyces scabies, S. ipomoeae, Pantoea agglomerans, Serratia marcescens, Streptomyces reticuliscabei, Acetobacter aceti, Spiroplasma citri Xylella fastidiosa, and/or Sphingomonas melonis.
As used herein, “disease resistance” or “disease tolerance” are used interchangeably and refer to a decrease in disease symptoms and/or a decrease pathogen growth and reproduction of a plant and/or part thereof. In some embodiments, the percent (%) increase in resistance/tolerance to disease as compared to a control may be in a range from about 0.1% to about 100%. In some embodiments, the percent increase in resistance/tolerance to disease may be an increase in a range from about 0.1% to about 10%, 0.1% to about 30%, about 0.1% to about 50%, about 0.1% to about 80%, about 0.1% to about 90%, about 0.1% to about 95%, about 1% to about 10%, about 1% to about 20%, about 1% to about 40%, about 1% to about 50%, about 1% to about 75%, about 1% to about 95%, about 1% to about 100%, about 10% to about 20%, about 10% to about 40%, about 10% to about 50%, about 10% to about 70%, about 10% to about 80%, about 10% to about 90%, about 10% to about 100%, about 20% to about 40%, about 20% to about 75%, about 20% to about 90%, about 20% to about 95%, about 20% to about 100%, about 25% to about 50%, about 50% to about 75%, about 50% to about 95%, about 50% to about 100%, about 75% to about 90%, about 75% to about 100%, about 90% to about 95%, about 90% to about 100% or any value or range therein, as compared to a control. In some embodiments, the % increase in resistance/tolerance to disease may be about 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17, 5, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, or 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%or 100% or any value or range therein, as compared to a control.
In some embodiments, a method for increasing abiotic stress tolerance of a plant and/or part thereof is provided, the method comprising applying a composition comprising an effective amount of MLGs to a plant and/or part thereof, and/or to the growth media, thereby increasing the abiotic stress tolerance of the plant and/or part thereof as compared to a control plant and/or part thereof (e.g., a plant and/or part thereof to which the composition comprising MLGs has not been applied). In some embodiments, the method comprises applying the composition at least one time (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times, or any range or value therein). In some embodiments, the method comprises applying the composition at least two times (e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times, or any range or value therein).
In some embodiments, abiotic stress may include, but is not limited to, drought, salinity (e.g., medium salinity (ECe=4-8 dSm-1); high salinity (ECe>8 dSm-1), flooding, freezing (e.g., about 0° C. or less), chilling or cold temperature (e.g., less than about 10-15° C.), heat or high temperatures (e.g., more than about 40° C.), high light intensity (e.g. more than about 10,000 foot candles), low light intensity (e.g. less than about 1000 foot candles), and/or ozone, and/or combinations thereof. In some embodiments, the abiotic stress is drought. In some embodiments, the abiotic stress is salinity.
As used herein, “an “increased tolerance to abiotic stress” or “increased resistance to abiotic stress” are used interchangeably and refer to the ability of a plant and/or part thereof,, where the plant and/or part thereof, and/or growth media has been contacted with a composition comprising MLGs, to withstand a given abiotic stress better than a control plant and/or part thereof (i.e., a plant and/or part thereof, and/or growth media, that has been exposed to the same abiotic stress but has not been contacted with a composition comprising MLGs). Increased tolerance to abiotic stress can be measured using a variety of parameters including, but not limited to, the size and number of plants and/or parts thereof, and the like (e.g., number and size of fruits), the level or amount of cell division, the amount of floral abortion, the amount of sunburn damage, crop yield, and the like. Thus, in some embodiments of this invention, a plant and/or part thereof, where the plant and/or part thereof, and/or growth media have been contacted with a composition comprising MLGs, having increased tolerance to the abiotic stress would have, for example, increased fruit/seed number and/or weight as compared to a plant and/or part thereof exposed to the same stress but not having been contacted with said composition.
In some embodiments, the percent increase in resistance/tolerance to abiotic stress as compared to a control may be an increase in a range from about 0.1% to about 100%. In some embodiments, the percent increase in resistance/tolerance to abiotic stress may be in a range from about 0.1% to about 10%, 0.1% to about 30%, about 0.1% to about 50%, about 0.1% to about 80%, about 0.1% to about 90%, about 0.1% to about 95%, about 1% to about 10%, about 1% to about 20%, about 1% to about 40%, about 1% to about 50%, about 1% to about 75%, about 1% to about 95%, about 1% to about 100%, about 10% to about 20%, about 10% to about 40%, about 10% to about 50%, about 10% to about 70%, about 10% to about 80%, about 10% to about 90%, about 10% to about 100%, about 20% to about 40%, about 20% to about 75%, about 20% to about 90%, about 20% to about 95%, about 20% to about 100%, about 25% to about 50%, about 50% to about 75%, about 50% to about 95%, about 50% to about 100%, about 75% to about 90%, about 75% to about 100%, about 90% to about 95%, about 90% to about 100% or any value or range therein, as compared to a control. In some embodiments, the percent increase in resistance/tolerance to abiotic stress may be about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17, 5, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, or 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% or any value or range therein, as compared to a control.
An embodiment of the present disclosure further provides methods for obtaining plants having increased fruit production compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having increased inflorescence production compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having increased fruit quality when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having increased production of defense/immunity-related calcium when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having increased expression of defense/immunity-related genes when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having increased tolerance to fungi when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining pepper plants having increased tolerance to white mold (Sclerotinia sclerotiorum) when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining cucumber plants having increased tolerance to Podosphaera fusca when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having increased tolerance to bacteria when treated with a composition comprising MLGs.
An embodiment of the present disclosure further provides methods for obtaining plants having increased tolerance to bacterial speck (Pseudomonas syringae) when treated with a composition comprising MLGs.
An embodiment of the present disclosure further provides methods for obtaining plants having increased tolerance to plant viruses when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining plants having decreased viral load when compared to plants where a composition comprising MLGs has not been applied.
An embodiment of the present disclosure further provides methods for obtaining zucchini plants having increased tolerance to tomato leaf curl New Dehli virus (ToLCNDV) compared to plants where a composition comprising MLGs has not been applied.
In some embodiments, a method for increasing nutrient use efficiency of a plant and/or part thereof is provided, the method comprising applying a composition comprising an effective amount of MLGs to a plant and/or plant part thereof, and/or to a growth media, thereby increasing nutrient use efficiency of the plant and/or part thereof as compared to a control plant and/or part thereof (e.g., a plant and/or part thereof to which the composition of the invention has not been applied). In some embodiments, the method comprises applying the composition at least once (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and/or 12 or more times, or any range or value therein). In some embodiments, the method comprises applying the composition at least two times (e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times, or any range or value therein).
In some embodiments, a method is provided for obtaining plants having increased fruit production (e.g., increased by about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein) compared to plants where a composition comprising MLGs has not been applied. Thus in some embodiments, the methods of the invention may provide plants having increased fruit production of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased inflorescence production (e.g., increased by about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein) compared to plants where a composition comprising MLGs has not been applied. Thus in some embodiments, the methods of the invention may provide a plant having increased inflorescence production of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased fruit quality when compared to plants where a composition comprising MLGs has not been applied (e.g., increase in fruit quality by about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein) compared to plants where a composition comprising MLGs has not been applied. Thus in some embodiments, the methods of the invention may provide a plant having increased fruit quality of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
An embodiment of the present disclosure further provides methods for obtaining plants having increased production of defense/immunity-related calcium when compared to plants where a composition comprising MLGs has not been applied (e.g., increased by about 1 fold to about 100 fold or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150 fold, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased production of defense/immunity-related calcium of about 1 to about 5 fold, about 1 to about 10 fold, about 1 to about 20 fold, about 5 to about 15 fold, about 5 to 20 fold, about 5 to about 25 fold, about 5 to about 30 fold, about 5 to about 50 fold, about 10 to about 20 fold, about 10 to about 30 fold, about 10 to about 50 fold, about 10 to about 70 fold, about 15 to about 20 fold, about 15% to about 50 fold, about 20 to about 30 fold, about 20 to about 50 fold, about 20 to about 70 fold, about 40 to about 50 fold, about 40 to about 80 fold, about 40 to about 100 fold, about 50 to about 70 fold, about 50 to about 100 fold, about 70 to about 100 fold, about 80 to about 100 fold, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased expression of defense/immunity-related genes when compared to plants where a composition comprising MLGs has not been applied (e.g., an increase of about 1 fold to about 100 fold or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150 fold, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased expression of defense/immunity-related genes of about 1 to about 5 fold, about 1 to about 10 fold, about 1 to about 20 fold, about 5 to about 15 fold, about 5 to 20 fold, about 5 to about 25 fold, about 5 to about 30 fold, about 5 to about 50 fold, about 10 to about 20 fold, about 10 to about 30 fold, about 10 to about 50 fold, about 10 to about 70 fold, about 15 to about 20 fold, about 15% to about 50 fold, about 20 to about 30 fold, about 20 to about 50 fold, about 20 to about 70 fold, about 40 to about 50 fold, about 40 to about 80 fold, about 40 to about 100 fold, about 50 to about 70 fold, about 50 to about 100 fold, about 70 to about 100 fold, about 80 to about 100 fold, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased tolerance to one or more fungal pathogens when compared to plants where a composition comprising MLGs has not been applied (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to one or more fungal pathogens of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining pepper plants having increased tolerance to white mold (Sclerotinia sclerotiorum) when compared to plants where a composition comprising MLGs has not been applied (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to white mold of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining cucumber plants having increased tolerance to Podosphaera fusca when compared to plants where a composition comprising MLGs has not been applied (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to P. fusca of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased tolerance to one or more pathogenic bacteria when treated with a composition comprising MLGs (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to one or more pathogenic bacteria of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased tolerance to bacterial speck (Pseudomonas syringae) when treated with a composition comprising MLGs (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to bacterial speck of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having increased tolerance to one or more plant pathogenic viruses when compared to plants where a composition comprising MLGs has not been applied (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to one or more plant pathogenic viruses of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, or about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining plants having a decreased viral load when compared to plants where a composition comprising MLGs has not been applied (e.g., a decrease of about 5% to about 100%; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100%, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having a decreased viral load of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, or about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
In some embodiments, a method is provided for obtaining zucchini plants having increased tolerance to tomato leaf curl New Dehli virus (ToLCNDV) compared to plants where a composition comprising MLGs has not been applied (e.g., an increase of about 5% to about 100% or more; e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150%, or more, or any range or value therein). Thus in some embodiments, the methods of the invention may provide a plant having increased tolerance to tomato leaf curl New Dehli virus (ToLCNDV) of about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 50%, 10% to about 70%, about 15% to about 20%, about 15% to about 30%, 15% to about 50%, about 20% to about 30%, about 20% to about 50%, about 20% to about 70%, about 40% to about 50%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 50% to about 70%, about 50% to about 100%, about 50% to about 125%, about 75% to about 100%, about 75% to about 120%, or about 75% to about 140%, and any range or value therein, as compared to a plant not contacted with a composition of the invention comprising MLGs.
The invention will now be described with reference to the following examples. It should be appreciated that these examples are not intended to limit the scope of the claims to the invention but are rather intended to be exemplary of certain embodiments. Any variations in the exemplified methods that occur to the skilled artisan are intended to fall within the scope of the invention.
Cellular calcium influx measured as relative luminescence units (RLU) over time in 8-days old Arabidopsis Col-0AEQ seedlings after treatment with
Results shown in
12-day-old Arabidopsis seedlings grown on liquid MS medium were treated with 50 µM MLG43 or water (mock) solutions for 0 and 30 minutes. Total RNA was purified with the RNeasy Plant Mini Kit (Qiagen) according to the manufacturer’s protocol.
For RNA-seq analyses, samples from three biological replicates for each treatment were selected and processed as previously described (Mélida et al., 2018). The incubation with MLG43, regulated the expression of 2062 genes, most of which (1375) were up-regulated as shown in Table 1 (see, end of Examples, pages 37-83). MLG43 up-regulated genes mainly grouped into GO terms related to immune system processes and to response to different stimuli, including biotic and abiotic as shown in annex Table 1.
Tomato plants var. Mayoral were sown in a greenhouse on Dec. 15, 2018 MLG43 at 0.05 g/l (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) was applied at three different rates by foliar application every three weeks for a total of 6 applications, starting one week after transplanting. Tomato fruits were harvested at 6 different time points in 2019. Total yield at each harvest was recorded. Fruit quality was assessed at harvest point 2, 4 and 6. Data represent the average +/-SD of 4 replicates distributed in random blocks with 40 plants /replicate. (** p<0.05, * P< 0.1).
As shown in Table 2, tomato plants treated with MLG43 exhibited an increase in total weight of fruit production of 9% when compared to untreated plants. Fruit size and weight was increased by 8% and 7%, respectively, in MLG43-treated tomato plants (Table 3).
Watermelon plants var. Motril and Boston were sown a greenhouse on Dec. 19, 2018. A total of 1600 plants were used, 800 control plants and 800 MLG43-treated plants distributed in random blocks. MLG43 -treated plants were sprayed four times once per month, from January to April 2019 with 0.05 g/l MLG43 (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) at 10 ml/plant/month. Control plants were treated with water or mock (adjuvant alone). The plants were harvested twice in April 2020 and production data were collected per harvest day according to the category, with category 1 (CAT1) being the best and category 2 (CAT2) being of less quality. Data represent the average +/-SD of 4 replicates distributed in random blocks with 40 plants /replicate. (** p<0.05).
As shown above in Table 4, watermelon plants treated with MLG43 display an increased total fruit production by 46.5% and increased fruit quality by 62% when compared to untreated plants. Moreover, only 5.2% of the MLG43-treated plants where of Category 2, compared to the 14.4% of the untreated plants.
Pepper plants var. California, Guepard and Ferrari were sown in a greenhouse on Dec. 19, 2018. A total of 512 plants were used per variety, 256 control plants and 256 MLG43-treated plants distributed in random blocks. MLG43 0.05 g/ (with adjuvants: surfactant 0.05%, antifoaming agent and biocide)-treated plants were sprayed 5 times, once per month from January to April 2019 with 0.05 g/l MLG43 (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) at 5ml/plant/month. Data represent the average +/-SD of 4 replicates distributed in random blocks with plants/replicate. (* P< 0.1).
As shown above in Table 5, pepper plants treated with MLG43 had an increase of fruit production between 22.5% and 33.7% compared to untreated plants.
As shown above in Table 6, pepper plants treated with MLG43 had an increase in the average weight of fruit between 3.48% and 14% compared to untreated plants.
Pepper plants var. Ferrari were sown on Nov. 26, 2018 and grown in a greenhouse. 0.125 g/l MLG43 (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) was applied by foliar spray at 2 ml/plant on 5 week old plants, 2 days prior inoculation with 5 ml/plant of a foliar spray of a 250 cfu/ml suspension of Sclerotinia sclerotiorum Plants were kept at 100% relative humidity for ten days and then humidity was lowered to 80% for the rest of the experiment. Disease symptoms were determined at 5 and 9 day-post inoculation (dpi) according to a disease severity index score where 0 corresponds to no symptoms and 4 corresponds to a dead leaf. Data represent the average +/-SD of 4 replicates distributed in random blocks with 6 plants /replicate. Different letters indicate statistically significant differences according to Student’s t-test (p<0.05).
As shown above in Table 7 the disease index in MLG43 treated pepper plants was significantly lower than in untreated pepper plants at 9 dpi.
Tomato plants (Solanum lycopersicum var. Moneymaker) were sown on Nov. 15, 2018 and grown in a greenhouse. Three-week-old tomato plants were sprayed with 2 ml of a 0.125 g/l MLG43 solution (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) or mock-treatment (surfactant 0.05%, antifoaming agent and biocide). Pseudomonas syringae DC3000 (108 cfu/ml) infection was performed 48 h after treatment with MLG43 solution or Mock. Tomato leaf discs were collected at 0- and 11-days post-inoculation (dpi) and colony forming units (cfu) per foliar area was determined. Data represent mean ± σ (n=8). Statistically significant differences according to Student’s t-test (** p<0.01).
As shown in
Cucumber plants were sown on and distributed in random blocks of 9 plants with 7 replicates (63 plants per treatment) and grown in a standard greenhouse between March and September 2019. MLG43 1 g/l (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) was sprayed at 1, 5, 10, 50, 100 and 200 g/ha 2 days before inoculation with Podosphaera fusca. Plants were then evaluated for 6, 9, and 14 days post inoculation (dpi) according to a disease severity index score where 0 corresponds to no symptoms and 5 corresponds to a dead plant. The area under the disease progress curve (AUDPC) was calculated as a quantitative summary of disease intensity over time as well as the efficacy of protection. Different letters indicate statistically significant differences according to Student’s t-test (p<0.05).
As shown above in Table 8 the disease index, AUDPC and efficacy in cucumber plants treated with six different rates of MLG43 were significantly lower than in untreated cucumber plants inoculated with Podosphaera fusca. The differences were dose dependent, the highest rate giving statistically the highest level of protection against pathogen disease. The EC50 value (rate inducing half-maximum resistance) was 50 g/ha.
Natural ToLCNDV infection occurred in zucchini plants var. Victoria and Cronos grown under standard greenhouse conditions by an experimented farmer in Almería, Spain. In total, 2200 (var. victoria) and 4300 (var. cronos) plants were used as untreated controls whereas 4400 (var. Victoria) and 2190 (var. Cronos) plants were sprayed every two weeks with MLG43 0.05 g/l (with adjuvants: surfactant 0.05%, antifoaming agent and biocide) in combination with abamectine twice and with either imidacloprod or spinosad alternating every other week. Furthermore, mancozeb was also added as a combination twice. Control plants were treated with abamectine or imidacloprod or spinosad or mancoceb alone. To detect viral load, completely randomized blocks of 10 plants with 18 replicates (180 plants for each control and MLG43-treated) were designed and 2 young leaves/plant were harvested and used for tissue print hybridization with a virus specific Digoxigenin-labeled probe on positively charged nylon membranes. The Digoxigenin-labeled probe was obtained by PCR amplification from the partial AV1 gene from DNA-A of ToLCNDV using the primer pairs ToNDA-580F:5′-TCACACATCGCGTAGGCAAG-3′ (SEQ ID NO:1) and ToNDA-935R: 5′-TGCCGGCCTCTTGTTGATTG-3′ (SEQ ID NO:2) with the PCR DIG Labeling Mix (Roche Diagnostics, Switzerland) and following the manufacturer’s instructions. Immunodetection was performed with anti-digoxigenin antibody conjugated with alkaline phosphatase (Roche Diagnostics, Switzerland) and chemiluminescence with CSPD (Roche Diagnostics, Switzerland) as substrate, following the manufacturer’s instructions and different times of exposure (15 min -overnight) to a Lumi-film (Amersham Bioscience, UK). Virus disease index was calculated as the number of TolCNDV positive plants/180 total number of sampled plants.
As shown above in Table 9, plants treated with MLG43 had a lower infection index by tomato leaf curl New Dehli virus (ToLCNDV). Table 10 below shows the total zucchini fruit production per plant in treated and untreated plants. Treated plants were sprayed every three weeks with 0.05 g/l MLG43.
Column 1 shows the location, column 2 shows the group, column 3 shows the average harvested production for each plant in kilograms, column 4 shows the number of plants for each group, column 5 shows the percentage of gain with respect to the control values, and column 6 shows the P-value corresponding to statistical T2 analysis for daily production per plant data and treatment. P-values lower than 0.05 indicate significant differences between control and treated groups (α=0.05). The plants were grown in a conventional production greenhouse in Almeria, Murcia and Granada, Spain. Percentage respect to the control values is shown for each year harvest data.
As shown above in Table 10, plants treated with MLG43 showed between 4.9% and 38.3% increased fruit production compared to untreated plants.
Five week old pepper plants (Capsicum annuum, Murano) were pre-treated with MLG43 as a foliar spray application with MLG43 (0.25 mg/plant) 2 days prior to Botrytis cinerea fungal inoculation. Control plants were mock treated. Two-days after treatment, Control and MLG43-treated plants were moved to a 75% humidity greenhouse chamber and spray-inoculated with 3 ml of Gamborg’s B5 medium containing B. cinerea conidia. Disease symptoms were determined at 5 dpi and 9 dpi in all the leaves of each plant (n = 12) using a scale from 0 to 4 where 0 = no symptoms; 1 = 9 little necrotic spots (< 10 % of leaf area); 2 = two or more notable necrotic spots (10 - 25 % of leaf area); 3 = big necrotic area (25 - 50 % of leaf area); 4 = more than 50 % of leaf area affected, 5 = leaf senescence. The MLG43-treated pepper plants showed a reduction in the disease symptoms index in comparison to mock-treated plants; the reduction was significant (p<0.05) both at 5 and 9 dpi(days post inoculation) for B. cinerea infections. The results are shown in
In a further study, pepper plants were treated with 0.125 mg/ml MLG43. In this study, pepper plants (C. annuum, Murano) were grown in a greenhouse in soil-vermiculite (3:1) at under 14 hours of light/10 hours of dark at 21-19° C. Five-weeks-old plants were mock treated (control) or treated using 2 ml of a MLG43 solution (0.125 mg/ml). Two-days after treatment, plants were moved to a 75% humidity greenhouse chamber and spray-inoculated with 3 ml of Gamborg’s B5 medium containing 106B. cinerea conidia (Benito et al., 1998). Disease symptoms were determined at 5 dpi and 9 dpi in all the leaves of each plant (n = 12) using a scale from 0 to 5 where 0 = no symptoms; 1 = little necrotic spots (< 10 % of leaf area); 2 = two or more notable necrotic spots (10 - 25 % of leaf area); 3 = big necrotic area (25 - 50 % of leaf area); 4 = more than 50 % of leaf area affected, 5 = leaf senescence. Disease symptoms index produced by Botrytis cinerea at 5 and 9 days post inoculation (dpi) in leaves of pepper plants is shown in
Treatment of Arabidopsis thaliana (Col-0 ecotype) with MLG43 confers enhanced disease resistance to the oomycete Hyaloperonospora arabidopsidis (Noco2 isolate). In this example, Arabidopsis thaliana (Col-0 ecotype) plants were grown in soil-vermiculite (3:1) under short day conditions (10 hours of light/14 hours of dark) at 21-20° C. and 75% humidity. Two-weeks-old plants were untreated (mock control) or treated by foliar spray using 0.1 ml of MLG43 solution per pot at either 0.1 mg/ml or 0.5 mg/ml. Two-days after treatment, plants were spray-inoculated with 0.1 ml conidia suspension (4·104 conidia/ml) of a H. arabidopsidis isolate Noco2. The presence of H. arabidopsidis in plants was quantified at 7 days post inoculation as the abundance of conidiospores/mg of plant fresh weight by harvesting the inoculated plants in water, shaking them and counting the conidiospores using a Neubauer chamber. The number of conidiospores of Hyaloperonospora arabidopsidis per mg of plant fresh weight determined at 7 days post inoculation showed increased disease resistance in the MLG43 treated plants. Statistically significant differences according to Student’s t-test (* p<0.05; ** p<0.01) were measured in the MLG43 treated plants compared to the mock treatment control plants. The results are shown in
Seeds from wheat (Triticum aestivum L.) cultivars Chinese Spring were sown in peat substrate and grown for 17 d in a glasshouse at 17° C. (day) and 15° C. (night) with a 16-h photoperiod and 60% humidity. For all infection experiments, 2×3 pot arrays (7×7 cm and 200 ml each) containing two seedlings per unit were used.
Z. tritici inoculum (Swiss strain ST99CH_3D7) (described by Zhan et al., Molecular Ecology, 14: 2683-2693 (2005)) was prepared as follows: 3D7 was inoculated in YPD (yeast extract 10 g/L, peptone 10 g/L, dextrose 20 g/L, agar 15 g/L) plates. After 4 d of incubation at 18° C., spores were collected in sterile deionised water and stored on ice until infection. The concentrations of the spore suspensions were determined with a Neubauer counting chambers and adjusted to 2.5×106spores ml-1 in 0.1% (v/v) for inoculation.
Wheat plants were mock treated (control) or sprayed with 12 ml of MLG43 at either 0.25 mg/ml or 0.75 mg/ml. Twenty four hours later plants were spray-inoculated with 12 ml of the spores suspension of the fungus. Pot arrays were placed into a sealing bag to keep humidity at 100% during 2 d post-inoculation. For symptom quantification, second leaves were mounted on paper sheets, scanned with a flatbed scanner (CanoScan LiDE 400) and analysed using automated image analysis (Stewart et al., Molecular Plant Pathology, 19: 201-216 (2016)). Data analysis and plotting was performed using RStudio v.1.0.143. Confidence intervals of the medians were determined using the ‘boot’ package and Kolmogorov-Smirnov (KS) tests for statistical significance with the ‘matching’ package in RStudio. Statistically significant differences in disease resistance were observed in MLG43 treatments compared with mock treated plants according to Kolmogorov-Smirnov (p-value<0.1). The results are shown in
The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.
| Number | Date | Country | Kind |
|---|---|---|---|
| 20382671.4 | Jul 2020 | EP | regional |
| 20382893.4 | Oct 2020 | EP | regional |
| 20383070.8 | Dec 2020 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2021/071085 | 7/27/2021 | WO |
