A frost mitigation method for the vegetative and flower buds of fruits using composite coating materials (CCM)

Abstract

This disclosure relates to a sophisticated method for protecting pome and stone fruit trees from freezing and subfreezing temperatures. The method involves applying a specialized Coating Composite Material (CCM), which can be used alone or in conjunction with other polymers, fertilizers, plant growth regulators (PGRs), and cryoprotectants, directly to the trees from the bud swell stage to full bloom. The protective CCM is primarily formulated from sodium alginate and calcium chloride, which together form a biodegradable film around plant tissues, providing the first layer of protection against frost. This advanced coating methodology not only forms a physical barrier against frost, but also enhances plant resilience to cold stress through a variety of biochemical mechanisms, offering a comprehensive solution to protect vulnerable fruit trees in changing climatic conditions.

Description

ATTORNEY DOCKET NO. 222204-2810A FROST MITIGATION METHOD FOR THE VEGETATIVE AND FLOWER BUDS OF POME AND STONE FRUITS USING COMPOSITE COATING MATERIALS (CCM) CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application Ser. No. 63 / 689,059, filed August 30, 2024, which is incorporated herein by reference in its entirety.BACKGROUND

[0002] The Intergovernmental Panel on Climate Change (IPCC) has detailed various potential outcomes for global warming by the 21st century's end, using Representative Concentration Pathways (RCPs) as a basis. These outcomes highlight that, contingent upon the rigor of measures to curb greenhouse gas emissions, we could see an increase in average global surface air temperatures ranging from 0.3°C to 4.8°C by 2100, relative to the recent historical period (1995-2014). Specifically, under a scenario of low emissions, SSP1-1.9, temperature rises could be between 0.2°C and 1.0°C, whereas a scenario of high emissions, SSP5-8.5, predicts a starker increase ranging from 2.4°C to 4.8°C. Other scenarios, including SSP1-2.6, SSP2-4.5, and SSP3-7.0, forecast varied temperature rises, thereby presenting a wide array of possible future conditions based on differing mitigation efforts (IPCC, 2021). Research increasingly shows that such a warming climate significantly affects temperate fruit species by altering crucial phenological traits (Fraga and Santos, 2021). This includes changes in the bud dormancy period, cold hardiness, blooming dates, which in turn affect tree productivity and sustainability. Dormancy, a survival mechanism for plants during winter to withstand freezing temperatures, involves inactivity and enhanced cold hardiness in plant meristem cells (Liu et al., 2021). Temperate fruit species have a specified winter chill requirement (CR) necessary to break dormancy and recommence growth in spring (Liu and Sherif, 2019). However, milder winters, as predicted under global warming, are expected to disrupt the completion of CR, leading to irregular and diminished crop yields as per IPCC projections (Fraga and Santos, 2021).

[0003] Further studies have underscored the profound impact of global climate change on the timing of plant dormancy and flowering stages, coupled with a heightened risk of spring frost events. Significant changes in plant phenology, including shorter dormancy periods and earlier budburst and flowering, have been recorded across a variety of woody perennials in the Northern Hemisphere (Augspurger, 2013; Ma et al., 2019). For example, over the past three decades, the flowering time of apple trees in Europe has advanced by approximately 6- 9 days (Hoffmann and Rath, 2013; Vitasse et al., 2011 ; Vitasse et al., 2018), a trend expected to continue in the future (Unterberger et al., 2018). Earlier bloom time in temperate tree fruit cultivars severely increases the risk and severity of late-spring frost and freeze damageATTORNEY DOCKET NO. 222204-2810(Lamichhane, 2021 ; Vitasse et al., 2018).

[0004] The distinction between "frost" and "freeze" is often misunderstood in various literature sources, mistakenly using the terms interchangeably. Frost occurs at air temperatures above the freezing point (>32°F) but with dew points below freezing, causing water vapor to condense as ice on surfaces, including plants, without necessarily freezing the water within the plant itself. Freezing takes place when air temperatures fall below the freezing point and dew points are significantly lower, presenting a distinct threat to plant tissues.

[0005] Plant susceptibility to frost or freeze damage depends on factors like species, genetic variety, and developmental stage. There are two primary types of frost trees face in spring: advective and radiation frosts, with radiation frosts being more common. Advective frosts are caused by a large cold air mass moving into an area and dropping air temperatures below freezing. Radiation frosts occur due to energy loss through radiant exchange at night under clear skies, non-windy conditions, and low dew points, leading to temperature inversions where air temperature drops below 32°F at night and rises above freezing during the day.

[0006] During a radiation frost event, interconnected processes significantly impact plant health, influenced by the rate and duration of temperature decline and the plant's inherent cold resistance mechanisms. Initially, ambient temperatures drop towards or below freezing, particularly under clear skies, low humidity, and calm winds at night, causing significant heat loss from the earth to the atmosphere. This leads to plant tissues cooling and undergoing supercooling, allowing water in plant tissues to remain liquid below the freezing point, effectively suppressing nucleation (Wisniewski et al., 2004; Londo and Kovaleski, 2019). However, supercooling provides only partial protection as progressive dehydration can still occur.

[0007] As temperatures continue to fall, extracellular ice may form, especially if freezing nuclei are present, drawing water out of cells and creating an osmotic gradient leading to cellular dehydration, volume loss, and potentially cell collapse or rupture (Pearce, 2001 ; Larcher, 2003; Snyder and de Melo-Abreu 2005; Zhang et al., 2019). To counteract lethal intracellular freezing, plants activate defense mechanisms, producing antifreeze proteins, sugars, and cryoprotectants to mitigate dehydration stress and prevent ice crystal growth within cells (Karabudak et al., 2014; Lunn et al., 2014; Judy and Kishore, 2016; Roychoudhury and Banerjee, 2016; Suo et al., 2017; Siddique et al., 2018; Liu et al., 2019; Chen et al., 2022). These adaptations also include morphological, biochemical, and physiological changes, such as osmolyte accumulation and cell wall reinforcement with suberin and lignin, to protect against freezing injury (Trache et al., 2017; Jian et al., 2020; Sun et al., 2021). However, rapid and severe extracellular freezing can lead to intracellular ice formation, often causing fatalATTORNEY DOCKET NO. 222204-2810 damage to cellular structures.

[0008] As dawn breaks and temperatures rise, ice thaws both externally and internally within plant cells. If the plant has successfully minimized internal ice formation, it may recover from the frost event. Yet, the thawing process can introduce risks, such as sudden changes in cell volume and osmotic pressure, potentially causing further stress or damage. The extent of frost damage varies widely and may not be immediately evident, ranging from wilted or discolored foliage to damaged flower buds and blossoms, underscoring the complexity of plant responses to frost and the critical need for effective protective measures.

[0009] The existing strategies for mitigating frost damage are broadly categorized into active and passive methods. Passive methods, considered pre-emptive, include the selection of coldhardy and / or late-bloom varieties, treatment with plant growth regulators and chemical substances to increase tolerance to freezing, and careful site selection for planting. Active methods, which are employed immediately before or after a frost event, encompass various mechanical and irrigation techniques designed to protect the crops from freezing temperatures. However, the effectiveness of these strategies is often contingent on specific weather conditions and can be compromised by factors such as wind speed and temperature inversions. Additionally, the high costs associated with implementing certain active methods, such as wind machines and heaters, limit their applicability and accessibility for many growers.

[0010] Despite advances in frost mitigation research for plants, a need still exists for an approach that aims not only to safeguard the buds, stems, and vegetative tissues of pome and stone fruits from freezing and subfreezing temperatures but also to address the economic and social ramifications of climate change-induced frost events on the fruit industry. By developing more resilient agricultural practices and technologies, the enhancement of the sustainability and productivity of fruit cultivation in the face of an increasingly unpredictable climate may be achieved. These needs and other needs are satisfied by the present disclosure.SUMMARY

[0011] In accordance with the purpose(s) of the disclosure, as embodied and broadly described herein, the disclosure, in one aspect, relates to compositions comprising sodium alginate and calcium chloride, creating a coating composite material (or “CCM”) that may be used to coat plants and enhances their protection against cold, freezing, and subfreezing temperatures.

[0012] Disclosed are compositions comprising: sodium alginate having a concentration; and calcium chloride having a concentration; wherein the concentration of the sodium alginate is 0.1 - 3 wt%.ATTORNEY DOCKET NO. 222204-2810

[0013] Also disclosed are methods for protecting plants from freezing and subfreezing temperatures, the method comprising the steps: applying a first composition on a surface of a plant via a first manner; and applying a second composition on the surface of the plant via a second manner; wherein the first composition comprises: sodium alginate; and at least one polymer; wherein the second composition comprises: calcium chloride; and wherein the first composition and the second composition combine to form a coating composite material.

[0014] Other systems, methods, features, and advantages of the present disclosure will be or become apparent to one with skill in the art upon examination of the following drawings and detailed description. It is intended that all such additional systems, methods, features, and advantages be included within this description, be within the scope of the present disclosure, and be protected by the accompanying claims. In addition, all optional and preferred features and modifications of the described aspects are usable in all aspects of the disclosure taught herein. Furthermore, the individual features of the dependent claims, as well as all optional and preferred features and modifications of the described aspects are combinable and interchangeable with one another.BRIEF DESCRIPTION OF THE FIGURES

[0015] Many aspects of the present disclosure can be better understood with reference to the following drawings. The components in the drawings are not necessarily to scale, emphasis instead being placed upon clearly illustrating the principles of the present disclosure. Moreover, in the drawings, like reference numerals designate corresponding parts throughout the several views.

[0016] FIGs. 1A-1 B show the use of a painting brush to apply high sodium alginate (SA) concentrations (3%), followed by a backpack sprayer application of calcium chloride (CaCI2) at 100 mM on cherry trees at the tight cluster stage.

[0017] FIGs. 1C-1 D show the application of a lower SA concentration (1.5%) using a pressurized sprayer, followed by CaCI2application.

[0018] FIG. 2A shows the application which involved treating trees with a 3% sodium alginate (SA) solution applied via a painting brush, followed by a spray of 100 mM calcium chloride (CaCI2) using a backpack sprayer at the tight-cluster stage. Photos were taken four days posttreatment.

[0019] FIG. 2B shows a comparison of fruit set rates between CCM-treated and untreated control trees two weeks following full bloom.

[0020] FIG. 3A shows the trees that were treated with sodium alginate (SA) at a concentration of 1.5% and calcium chloride (CaCI2) at 100 mM at the tight-cluster stage.ATTORNEY DOCKET NO. 222204-2810Photos were taken for the same cluster at 2, 4 and 8 days after treatment (DAT).

[0021] FIG. 3B shows a comparison of fruit set rates between CCM-treated and untreated control trees two weeks following full bloom.

[0022] FIGs. 4A-4B shows branches of 'Sunhigh' peach trees at the pink stage, comparing untreated (FIG. 4A) to those treated with 1 .5% CCM (FIG. 4B) the day prior.

[0023] FIGs. 4C-4D shows longitudinal sections of these flower buds. Red arrow indicates brown or dead ovaries, while green arrows highlight green or alive flowers.

[0024] FIG. 5 shows a bar graph that presents the average findings from an assessment of 50 flower buds randomly selected from 'Sunhigh' peach branches. The buds were evaluated for flower mortality six hours following a frost event, illustrating the protective effects of the CCM coating.

[0025] FIG. 6 shows the delayed flowering observed in peach 'Sunhigh' trees treated with a 1 .5% CCM coating compared to untreated control trees. Photographs were captured six days following the CCM application, during which no rainfall occurred that might have washed off the coating film.

[0026] FIG. 7 shows the effect of sodium alginate concentration on bud mortality of ‘Red Delicious’ apple trees following a frost event. Bars represent mean percentage mortality (± SE) for king flowers (left bars) and side flowers (right bars) across five treatments: 0.50% sodium alginate, 0.25% sodium alginate, 0.10% sodium alginate, untreated control, and control with urea only. Buds were collected approximately six hours after the frost event, and mortality was determined based on ovary and style tissue browning. Different letters above bars indicate statistically significant differences among treatments within each flower type according to Tukey’s HSD test at P < 0.05.

[0027] Additional advantages of the disclosure will be set forth in part in the description which follows, and in part will be obvious from the description, or can be learned by practice of the disclosure. The advantages of the disclosure will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure, as claimed.DETAILED DESCRIPTION

[0028] Many modifications and other aspects disclosed herein will come to mind to one skilled in the art to which the disclosed compositions and methods pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the disclosures are not to be limited to the specificATTORNEY DOCKET NO. 222204-2810 aspects disclosed and that modifications and other aspects are intended to be included within the scope of the appended claims. The skilled artisan will recognize many variants and adaptations of the aspects described herein. These variants and adaptations are intended to be included in the teachings of this disclosure and to be encompassed by the claims herein.

[0029] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0030] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual aspects described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several aspects without departing from the scope or spirit of the present disclosure.

[0031] Any recited method can be carried out in the order of events recited or in any other order that is logically possible. That is, unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order. Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is no way intended that an order be inferred, in any respect. This holds for any possible non-express basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0032] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided herein can be different from the actual publication dates, which can require independent confirmation.

[0033] While aspects of the present disclosure can be described and claimed in a particular statutory class, such as the system statutory class, this is for convenience only and one of skill in the art will understand that each aspect of the present disclosure can be described and claimed in any statutory class.

[0034] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed compositions and methods belong. It will be further understood that terms, such as those defined in commonly usedATTORNEY DOCKET NO. 222204-2810 dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly defined herein.

[0035] Prior to describing the various aspects of the present disclosure, the following definitions are provided and should be used unless otherwise indicated. Additional terms may be defined elsewhere in the present disclosure.A. DEFINITIONS

[0036] As used herein, “comprising” is to be interpreted as specifying the presence of the stated features, integers, steps, or components as referred to, but does not preclude the presence or addition of one or more features, integers, steps, or components, or groups thereof. Moreover, each of the terms “by”, “comprising,” “comprises”, “comprised of,” “including,” “includes,” “included,” “involving,” “involves,” “involved,” and “such as” are used in their open, non-limiting sense and may be used interchangeably. Further, the term “comprising” is intended to include examples and aspects encompassed by the terms “consisting essentially of’ and “consisting of.” Similarly, the term “consisting essentially of’ is intended to include examples encompassed by the term “consisting of.

[0037] As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items. Expressions such as “at least one of,” when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

[0038] As used herein, nomenclature for compounds, including organic compounds, can be given using common names, IUPAC, IUBMB, or CAS recommendations for nomenclature. When one or more stereochemical features are present, Cahn-lngold-Prelog rules for stereochemistry can be employed to designate stereochemical priority, E / Z specification, and the like. One of skill in the art can readily ascertain the structure of a compound if given a name, either by systemic reduction of the compound structure using naming conventions, or by commercially available software, such as CHEMDRAW™ (Cambridgesoft Corporation, U.S.A.).

[0039] Reference to "a" chemical compound refers to one or more molecules of the chemical compound rather than being limited to a single molecule of the chemical compound. Furthermore, the one or more molecules may or may not be identical, so long as they fall under the category of the chemical compound. Thus, for example, "a" chemical compound is interpreted to include one or more molecules of the chemical, where the molecules may or may not be identical (e.g., different isotopic ratios, enantiomers, and the like).ATTORNEY DOCKET NO. 222204-2810

[0040] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a polymer,” “an alcohol,” or “a catalyst,” includes, but is not limited to, two or more such polymers, alcohols, or catalysts, and the like.

[0041] Reference to "a / an" chemical compound, protein, and antibody each refers to one or more molecules of the chemical compound, protein, and antibody rather than being limited to a single molecule of the chemical compound, protein, and antibody. Furthermore, the one or more molecules may or may not be identical, so long as they fall under the category of the chemical compound, protein, and antibody. Thus, for example, "an" antibody is interpreted to include one or more antibody molecules of the antibody, where the antibody molecules may or may not be identical (e.g., different isotypes and / or different antigen binding sites as may be found in a polyclonal antibody).

[0042] It should be noted that ratios, concentrations, amounts, and other numerical data can be expressed herein in a range format. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms a further aspect. For example, if the value “about 10” is disclosed, then “10” is also disclosed.

[0043] When a range is expressed, a further aspect includes from the one particular value and / or to the other particular value. For example, where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure, e.g. the phrase “x to y” includes the range from ‘x’ to ‘y’ as well as the range greater than ‘x’ and less than ‘y’. The range can also be expressed as an upper limit, e.g. ‘about x, y, z, or less’ and should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘less than x’, less than y’, and ‘less than z’. Likewise, the phrase ‘about x, y, z, or greater’ should be interpreted to include the specific ranges of ‘about x’, ‘about y’, and ‘about z’ as well as the ranges of ‘greater than x’, greater than y’, and ‘greater than z’. In addition, the phrase “about ‘x’ to ‘y’”, where ‘x’ and ‘y’ are numerical values, includes “about x’ to about ‘y’”.

[0044] It is to be understood that such a range format is used for convenience and brevity, and thus, should be interpreted in a flexible manner to include not only the numerical valuesATTORNEY DOCKET NO. 222204-2810 explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. To illustrate, a numerical range of “about 0.1 % to 5%” should be interpreted to include not only the explicitly recited values of about 0.1 % to about 5%, but also include individual values (e.g., about 1 %, about 2%, about 3%, and about 4%) and the sub-ranges (e.g., about 0.5% to about 1.1 %; about 5% to about 2.4%; about 0.5% to about 3.2%, and about 0.5% to about 4.4%, and other possible sub-ranges) within the indicated range.

[0045] As used herein, the terms “about,” “approximate,” “at or about,” and “substantially” mean that the amount or value in question can be the exact value or a value that provides equivalent results or effects as recited in the claims or taught herein. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art such that equivalent results or effects are obtained. In some circumstances, the value that provides equivalent results or effects cannot be reasonably determined. In such cases, it is generally understood, as used herein, that “about” and “at or about” mean the nominal value indicated ±10% variation unless otherwise indicated or inferred. In general, an amount, size, formulation, parameter or other quantity or characteristic is “about,” “approximate,” or “at or about” whether or not expressly stated to be such. It is understood that where “about,” “approximate,” or “at or about” is used before a quantitative value, the parameter also includes the specific quantitative value itself, unless specifically stated otherwise.

[0046] As used herein, the term “effective amount” refers to an amount that is sufficient to achieve the desired modification of a physical property of the composition or material. For example, an “effective amount” of sodium alginate refers to an amount that is sufficient to achieve the desired improvement in the property modulated by the formulation component, e.g. achieving the desired level of modulus. The specific level in terms of wt% in a composition required as an effective amount will depend upon a variety of factors including the amount and type of sodium alginate, amount and type of calcium chloride, amount and type of polymer, and amount and type of other additives and the like.

[0047] As used herein, the terms “optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0048] Unless otherwise specified, temperatures referred to herein are based on atmospheric pressure (i.e. one atmosphere).ATTORNEY DOCKET NO. 222204-2810B. COMPOSITIONS

[0049] Primary components of the CCM and their possible concentrations may comprise Sodium Alginate (0.1 % to 3%) and Calcium Chloride (5-100 mM): These components are known for their ability to form a biodegradable film around plant tissues, serving as the initial layer of protection against frost.

[0050] In addition to these components, the CCM may contain one or more of the following along with their possible concentrations as enhanced protective additives:

[0051] Xanthan Gum (0.1% to 1 %): May increase the viscosity and adhesion of the coating, prolonging its protective effect and enhancing resistance to environmental factors.

[0052] Antifreeze Proteins (AFPs) (1 to 10 mg / L): May improve frost resistance by inhibiting ice crystal growth within plant tissues, thus reducing cellular damage.

[0053] Humectants (Glycerol 1-5%, Sorbitol 1-5%, Propylene Glycol 1-5%): These compounds can help retain moisture within the bud, mitigating desiccation stress under frost conditions.

[0054] Polyvinyl Alcohol (PVA) (0.1 % to 5%): Can be mixed with sodium alginate to form a more flexible, durable film, possibly improving the coating's resistance to physical damage and wear.

[0055] Chitosan (0.1 % to 2%): May enhance the mechanical strength and barrier properties of the alginate film with its film-forming, biodegradable, and antimicrobial properties.

[0056] Nanomaterials (Cellulose Nanocrystals, Silica Nanoparticles) (0.1 % to 3%): Act as reinforcing agents within the polymer matrix, possibly improving rigidity and strength without significantly increasing thickness or weight.

[0057] Ethylene Vinyl Acetate (EVA) (0.1 % to 10%): Can modify the coating's flexibility and water resistance, possibly aiding in the formation of a semi-permeable barrier that supports gas exchange while ensuring frost protection.

[0058] Cross-linking Enhancers (Magnesium, Barium) (1-50 mM): May enhance the crosslinking efficiency between sodium alginate and calcium ions, promoting a denser gel network.

[0059] Essential Oils (0.01% to 0.1 %): Can offer antifreeze and antimicrobial benefits, with careful optimization required to avoid phytotoxic effects.

[0060] Micronutrients (Zinc, Manganese, Boron) (1 to 100 ppm): Can support bud health and enhance resistance to stress conditions, including frost.

[0061] Seaweed Extract (0.1% to 3%): Rich in bioactive compounds, minerals, and growthATTORNEY DOCKET NO. 222204-2810 hormones, seaweed extract promotes plant vigor, enhances stress resistance including frost tolerance, and supports overall plant health.

[0062] Protein Additives (Gelatin, Whey Protein Isolate) (0.1% to 5%): Can improve the strength and flexibility of alginate films, possibly allowing for lower alginate concentrations while maintaining or improving the film's rigidity.

[0063] Plant Growth Regulators (PGRs) and Hormone Biosynthetic Inhibitors: ethylene Inhibitors (e.g., 1 -Methylcyclopropene, 1-MCP) (1-100 ppm): By inhibiting ethylene, these compounds can help maintain cell integrity and delay senescence in response to stress; abscisic Acid (ABA) (1-500 pM): Plays a critical role in plant response to environmental stress, including cold; ABA can improve water retention and induce the expression of cold-responsive genes; jasmonic acid (JA) Inhibitors (e.g, DIECA, Naproxen) (10-500 pM): While jasmonic acid is involved in stress responses, its inhibitors can modulate the plant's stress response to avoid overreaction, helping to maintain energy reserves for recovery.

[0064] The CCM may also comprise polymers including: polyvinyl alcohol, chitosan, polyethylene, polypropylene, polystyrene, polyacrylamide, polyacrylic acid, cellulose, starch, collagen, and gelatin.C. PLANT BENEFICIARIES OF CCM APPLICATION

[0065] Possible species that could benefit from the CCM application may include, but is not limited to: Cherry (Prunus); Sweet Cherry (Prunus avium); Sour Cherry (Prunus cerasus); Peach (Prunus persica), including Nectarines (often classified as Prunus persica var. nucipersica); Plum (Prunus domestica), including various subspecies and hybrids; Apricot (Prunus armeniaca); Apple (Malus domestica); Pear (Pyrus); Common Pear (Pyrus communis); Asian Pear (Pyrus pyrifolia); Quince (Cydonia oblonga); Almond (Prunus dulcis); Walnut (Juglans regia); Pecan (Carya illinoinensis); Hazelnut (Corylus avellana); Fig (Ficus carica); Persimmon (Diospyros); American Persimmon (Diospyros virginiana); Japanese Persimmon (Diospyros kaki); Pawpaw (Asimina triloba); Mulberry (Morus); Red Mulberry (Morus rubra); White Mulberry (Morus alba); Jujube (Ziziphus jujuba); and Medlar (Mespilus germanica).D. CCM’s APPLICATION1. CCM APPLICATION TIMING

[0066] In the life cycle of deciduous fruit trees like pome and stone fruits, the phenological stages from bud swell to full bloom are critical for successful fruit development, providing optimal windows for applying protective coatings. The application of a protective coating from the first swell stage to full bloom can shield these delicate stages from frost. Depending on theATTORNEY DOCKET NO. 222204-2810 coating's strength, thickness, and composition, it may also slow the transition from one stage to the next. This deceleration can be critical for frost protection as it keeps the buds at less vulnerable stages for longer periods. Importantly, the critical (lethal) temperature that can cause damage to flower and vegetative buds increases as the buds progress toward full bloom. Therefore, a well-timed application not only protects but can optimize the developmental trajectory against climatic risks.2. CCM APPLICATION METHODOLOGY

[0067] For the effective application of the CCM materials on fruit trees to protect against frost, there are two primary methods depending on the concentration of the coating solution: (1) Manual Application and (2) Sprayer Application.

[0068] For higher concentrations (above 1 wt%) of the coating material, manual application methods such as brushes, rollers, or sponges may be used. These tools allow for precise control over the application, ensuring that a thicker, more concentrated layer of the coating is evenly distributed over the buds and branches. This method is useful for small-scale applications or in research settings where specific areas of a tree need targeted protection.

[0069] For lower concentrations of the coating material, various types of sprayers can be used to ensure a uniform and widespread application across an orchard. This method is ideal for commercial growers due to its efficiency and effectiveness over large areas. Examples of sprayers commonly used in orchards may include: Backpack Sprayers (portable and suitable for small to medium-sized orchards or for targeted application, allowing the operator to control the pressure and coverage closely); Handheld Sprayers (similar to backpack sprayers but typically smaller, these are ideal for very small orchards or individual trees); Airblast Sprayers (also known as mist blowers, these sprayers are highly efficient for larger orchards, using a powerful air stream to carry droplets to the target, ensuring good coverage and penetration through the canopy); Boom Sprayers (mounted on tractors, boom sprayers can cover wide rows efficiently, and they may feature a long arm (boom) with multiple nozzles that spray the coating material as the tractor moves through the orchard); Electrostatic Sprayers (these sprayers charge the coating droplets as they are emitted, causing them to wrap around and uniformly coat all surfaces of the plant material, and ensuring thorough coverage, even on hard-to-reach surfaces); and Droplet-Control Sprayers (these sprayers are designed to minimize drift by producing larger droplets that are less likely to be carried away by wind, making them particularly useful in windy conditions or when precision is crucial to avoid contaminating nearby crops).

[0070] In both manual and spray application methods, the initial coating, which contains sodium alginate, may be applied first. Before this layer has a chance to dry completely, aATTORNEY DOCKET NO. 222204-2810 second coating containing calcium chloride, or other cross-linking materials may be sprayed over it. This process facilitates the formation of a film around the target tissues, effectively encapsulating them in a protective layer.E. REMOVAL OF THE CCM COATING

[0071] When the need arises to remove the CCM protective coating from fruit trees, it is important to use a method that is both effective and safe for the plant, while ensuring no harmful residues are left behind. The dissolution of this biodegradable film is achieved using specific chelating and disintegrating agents that break down the cross-linked structure of the coating.

[0072] A dissolving solution, or dissolution, may be used for the removal of the CCM coating. This solution may contain one or more of the following reagents, along with their possible concentrations: Sodium Citrate (1 to 100 mM), Sodium Hexametaphosphate (1 to 100 mM), EDTA (Ethylenediaminetetraacetic Acid) (1 to 100 mM), Sodium Carbonate and Sodium Bicarbonate (1 to 100 mM).

[0073] For method of application, the chosen dissolution agent is diluted in water to form a solution, which should be applied using a sprayer similar to those used for the initial coating application. This ensures even coverage and effective penetration of the solution into the film.

[0074] The dissolution solution may be applied when the tree is dry and during cooler parts of the day to minimize evaporation and maximize the solution's impact.

[0075] All chosen agents (sodium citrate, sodium hexametaphosphate, EDTA, sodium carbonate, and sodium bicarbonate) are generally recognized as safe for use in agricultural applications. However, appropriate concentrations will be maintained to prevent any potential phytotoxic effects.

[0076] By using these materials and methods, the removal process of the sodium alginatecalcium chloride coating is likely not only effective but also safe for the trees and the environment. This approach ensures that the protective benefits of the coating can be reversed easily likely without leaving any detrimental residues, maintaining the health and productivity of the orchard.F. ASPECTS

[0077] The following listing of exemplary aspects supports and is supported by the disclosure provided herein.

[0078] Aspect 1. A composition, comprising: (a) sodium alginate having a concentration; and (b) calcium chloride having a concentration; wherein the concentration of the sodium alginateATTORNEY DOCKET NO. 222204-2810 is 0.1-3 wt%.

[0079] Aspect 2. The composition of Aspect 1 , wherein the concentration of the sodium alginate is 0.5-1 .5 wt%.

[0080] Aspect 3. The composition of Aspect 1 , wherein the concentration of the calcium chloride is 5-100 mM.

[0081] Aspect 4. The composition of Aspect 1 , wherein the concentration of the calcium chloride is 5-50 mM.

[0082] Aspect 5. The composition of any one of Aspects 1-4, wherein the composition further comprises one or more of Xanthan gum, antifreeze proteins, glycerol, sorbitol, propylene glycol, polyvinyl alcohol, chitosan, cellulose nanocrystals, silica nanoparticles, ethylene vinyl acetate, magnesium, barium, oils, zinc, manganese, boron, seaweed extract, gelatin, whey protein isolate, plant growth regulators, hormone biosynthetic inhibitors, ethylene inhibitors, abscisic acid, and jasmonic acid inhibitors.

[0083] Aspect 6. The composition of Aspect 5, wherein the composition further comprises Xanthan gum, wherein the Xanthan gum has a concentration of 0.1-1 wt%.

[0084] Aspect 7. The composition of Aspect 5, wherein the composition further comprises glycerol, wherein the glycerol has a concentration of 1-5 wt%.

[0085] Aspect 8. The composition of Aspect 5, wherein the composition further comprises sorbitol, wherein the sorbitol has a concentration of 1-5 wt%.

[0086] Aspect 9. The composition of Aspect 5, wherein the composition further comprises propylene glycol, wherein the propylene glycol has a concentration of 1-5 wt%.

[0087] Aspect 10. The composition of Aspect 5, wherein the composition further comprises polyvinyl alcohol, wherein the polyvinyl alcohol has a concentration of 0.1-5 wt%.

[0088] Aspect 11 . The composition of Aspect 5, wherein the composition further comprises chitosan, wherein the chitosan has a concentration of 0.1-2 wt%.

[0089] Aspect 12. The composition of Aspect 5, wherein the composition further comprises silica nanoparticles, wherein the silica nanoparticles have a concentration of 0.1-3 wt%.

[0090] Aspect 13. The composition of Aspect 5, wherein the composition further comprises cellulose nanocrystals, wherein the cellulose nanocrystals have a concentration of 0.1-3 wt%.

[0091] Aspect 14. The composition of Aspect 5, wherein the composition further comprises ethylene vinyl acetate, wherein the ethylene vinyl acetate has a concentration of 0.1-10 wt%.

[0092] Aspect 15. The composition of Aspect 5, wherein the composition further comprisesATTORNEY DOCKET NO. 222204-2810 magnesium, wherein the magnesium has a concentration of 1-50 mM.

[0093] Aspect 16. The composition of Aspect 5, wherein the composition further comprises oils, wherein the oils have a concentration of 0.01-0.1 wt%.

[0094] Aspect 17. The composition of Aspect 5, wherein the composition further comprises zinc, wherein the zinc has a concentration of 1-100 ppm.

[0095] Aspect 18. The composition of Aspect 5, wherein the composition further comprises manganese, wherein the manganese has a concentration of 1-100 ppm.

[0096] Aspect 19. The composition of Aspect 5, wherein the composition further comprises boron, wherein the boron has a concentration of 1-100 ppm.

[0097] Aspect 20. The composition of Aspect 5, wherein the composition further comprises seaweed extract, wherein the seaweed extract has a concentration of 0.1-3 wt%.

[0098] Aspect 21. The composition of Aspect 5, wherein the composition further comprises gelatin, wherein the gelatin has a concentration of 0.1-5 wt%.

[0099] Aspect 22. The composition of Aspect 5, wherein the composition further comprises whey protein isolate, wherein the whey protein isolate has a concentration of 0.1-5 wt%.

[0100] Aspect 23. The composition of Aspect 5, wherein the composition further comprises an ethylene inhibitor, wherein the ethylene inhibitor has a concentration of 1-100 ppm.

[0101] Aspect 24. The composition of Aspect 5, wherein the composition further comprises abscisic acid, wherein the abscisic acid has a concentration of 1-500 pM.

[0102] Aspect 25. The composition of Aspect 5, wherein the composition further comprises a jasmonic acid inhibitor, wherein the jasmonic acid inhibitor has a concentration of 10-500 pM.

[0103] Aspect 26. The composition of any one of Aspects 1-25, wherein the composition is used to protect plants from freezing and subfreezing temperatures.

[0104] Aspect 27. A method for protecting plants from freezing and subfreezing temperatures, the method comprising the steps: (a) applying a first composition on a surface of a plant via a first manner; and (b) applying a second composition on the surface of the plant via a second manner; wherein the first composition comprises: sodium alginate; and at least one polymer; wherein the second composition comprises: calcium chloride; and wherein the first composition and the second composition combine to form a coating composite material.

[0105] Aspect 28. The method of Aspect 27, wherein the at least one polymer is selected from polyvinyl alcohol, chitosan, polyethylene, polypropylene, polystyrene, polyacrylamide,ATTORNEY DOCKET NO. 222204-2810 polyacrylic acid, cellulose, starch, collagen, and gelatin.

[0106] Aspect 29. The method of Aspect 28, wherein the at least one polymer is polyvinyl alcohol.

[0107] Aspect 30. The method of Aspect 28, wherein the at least one polymer is chitosan.

[0108] Aspect 31. The method of Aspect 29, wherein the polyvinyl alcohol has a concentration of 0.1-5 wt%.

[0109] Aspect 32. The method of Aspect 30, wherein the chitosan has a concentration of 0.5- 2 wt%.

[0110] Aspect 33. The method of any one of Aspects 27-32, wherein the first manner is manual application, using one or more of a brush, a roller, or a sponge.

[0111] Aspect 34. The method of any one of Aspects 27-32, wherein the first manner is application by a sprayer.

[0112] Aspect 35. The method of any one of Aspects 27-32, wherein the second manner is application by a sprayer.

[0113] Aspect 36. The method of any one of Aspects 27-35, wherein the method further comprises a step for removing the coating composite material from the plant, wherein a dissolving solution is applied to the plant.

[0114] Aspect 37. The method of Aspect 36, wherein the dissolving solution comprises one or more of sodium citrate, sodium hexametaphosphate, ethylenediaminetetraacetic acid, sodium carbonate, and sodium bicarbonate.

[0115] Aspect 38. The method of any one of Aspects 27-37, wherein the plant is selected from cherry, peach, plum, apricot, apple, pear, and quince.

[0116] From the foregoing, it will be seen that aspects herein are well adapted to attain all the ends and objects hereinabove set forth together with other advantages which are obvious and which are inherent to the structure.

[0117] While specific elements and steps are discussed in connection to one another, it is understood that any element and / or steps provided herein is contemplated as being combinable with any other elements and / or steps regardless of explicit provision of the same while still being within the scope provided herein.

[0118] It will be understood that certain features and subcombinations are of utility and may be employed without reference to other features and subcombinations. This is contemplated by and is within the scope of the claims.ATTORNEY DOCKET NO. 222204-2810

[0119] Since many possible aspects may be made without departing from the scope thereof, it is to be understood that all matter herein set forth or shown in the accompanying drawings and detailed description is to be interpreted as illustrative and not in a limiting sense.

[0120] It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting. The skilled artisan will recognize many variants and adaptations of the aspects described herein. These variants and adaptations are intended to be included in the teachings of this disclosure and to be encompassed by the claims herein.

[0121] Now having described the aspects of the present disclosure, in general, the following Examples describe some additional aspects of the present disclosure. While aspects of the present disclosure are described in connection with the following examples and the corresponding text and figures, there is no intent to limit aspects of the present disclosure to this description. On the contrary, the intent is to cover all alternatives, modifications, and equivalents included within the spirit and scope of the present disclosure.G. EXAMPLES

[0122] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated, and are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric.1. EXPERIMENT 1 : EFFICACY OF 3% COM ON SWEET CHERRY TREES

[0123] In this experiment, a 3% sodium alginate (SA) solution was applied to select branches of mature sweet cherry trees located at the Alson H. Smith Agricultural Research and Extension Center (AHS AREC) in Winchester, VA. The solution was brushed onto 2-3 flagged branches per tree. The remaining branches served as untreated controls. Subsequently, a 100 mM calcium chloride (CaCh) solution was sprayed onto the SA-treated branches using a backpack sprayer (FIGs. 1A-1 B). Treatments were made on March 15, 2024. Fruit set was assessed as the percentage of flowers that developed into fruit relative to the total number of initial flowers, with data collected from 20 flower clusters on both treated and untreated branches two weeks following full bloom (FIG. 2).2. EXPERIMENT 2: EFFICACY OF 1.5% CCM ON S EET CHERRY TREESATTORNEY DOCKET NO. 222204-2810

[0124] This experiment mirrored the conditions of Experiment 1 but utilized a 1 .5% SA concentration. SA was sprayed onto half of each tree's canopy using a tractor-mounted pressurized sprayer, with the other half serving as a control. While the SA was still wet, the CaCh solution was applied using a backpack sprayer (FIGs. 1C-1 D). Treatments for this experiment were made on March 21 , 2024. The trees were at the tight-cluster stage of flower development during treatments. T reated branches showed signs of bloom delay compared to untreated controls (FIG. 3A). Fruit set percentages were calculated as in Experiment 1 (FIG. 3B).3. EXPERIMENT 3: EFFICACY OF 1.5% COM ON PEACH TREES

[0125] Mature ‘Sunhigh’ peach trees at the Alson H. Smith Agricultural Research and Extension Center (AHS AREC) received treatments similar to those in Experiment 2, with applications made about 12 hours prior to a forecasted frost event. Approximately six hours after the frost, 50 flower buds were collected from both CCM-treated and untreated trees to assess mortality. The flowers were dissected using a razor blade; buds with brown ovary and style tissues were deemed dead, while those with green tissues were classified as alive (refer to FIG. 4). Flower mortality was quantified by calculating the ratio of dead flowers to the total number of flowers examined (refer to FIG. 5). Notably, CCM treatments were applied when the trees were at the Pink stage of flower development. Additionally, peach trees treated with 1.5% CCM that was not washed off by rain exhibited a delay in blooming compared to untreated controls (refer to FIG. 6).4. EXPERIMENT 4: EFFICACY OF CCM APPLICATIONS ON ‘RED DELICIOUS’ APPLE TREES

[0126] Mature ‘Red Delicious’ apple trees at the Alson H. Smith Agricultural Research and Extension Center (AHS AREC) received foliar applications of CCM at concentrations of 0.50%, 0.25%, and 0.10%, in addition to two control treatments: untreated control and control with urea only. Applications were made approximately 12 hours prior to a forecasted frost event. Approximately six hours after the frost, 50 flower buds were collected from each treatment group to assess flower mortality.

[0127] The flowers were dissected using a razor blade; buds with brown ovary and style tissues were deemed dead, while those with green tissues were classified as alive. Flower mortality was quantified by calculating the ratio of dead flowers to the total number of flowers examined.

[0128] As shown in FIG. 7, CCM at 0.50% and 0.25% concentrations significantly reduced king flower mortality compared to both controls, with mortality rates of approximately 77% and 86%, respectively, versus ~100% in the untreated and urea controls. Side flower mortality was lowest in the 0.25% treatment (~56%), followed closely by the 0.50% treatment (~60%), bothATTORNEY DOCKET NO. 222204-2810 of which were significantly lower than the untreated control (-80%) and urea control (-72%). The 0.10% treatment resulted in intermediate side flower mortality (-74%), not significantly different from the controls.

[0129] It will be apparent to those skilled in the art that various modifications and variations can be made in the present disclosure without departing from the scope or spirit of the disclosure. Other aspects of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the disclosure disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the disclosure being indicated by the following claims.

Claims

ATTORNEY DOCKET NO. 222204-2810CLAIMSWhat is claimed is:1 . A composition, comprising: sodium alginate having a concentration; and calcium chloride having a concentration; wherein the concentration of the sodium alginate is 0.1-3 wt%.

2. The composition of claim 1 , wherein the concentration of the sodium alginate is 0.5- 1.5 wt%.

3. The composition of claim 1 , wherein the concentration of the calcium chloride is 5- 100 mM.

4. The composition of claim 1 , wherein the concentration of the calcium chloride is 5-50 mM.

5. The composition of claim 1 , wherein the composition further comprises one or more of Xanthan gum, antifreeze proteins, glycerol, sorbitol, propylene glycol, polyvinyl alcohol, chitosan, cellulose nanocrystals, silica nanoparticles, ethylene vinyl acetate, magnesium, barium, oils, zinc, manganese, boron, seaweed extract, gelatin, whey protein isolate, plant growth regulators, hormone biosynthetic inhibitors, ethylene inhibitors, abscisic acid, and jasmonic acid inhibitors.

6. The composition of claim 5, wherein the composition further comprises Xanthan gum, wherein the Xanthan gum has a concentration of 0.1-1 wt%.

7. The composition of claim 5, wherein the composition further comprises glycerol, wherein the glycerol has a concentration of 1-5 wt%.

8. The composition of claim 5, wherein the composition further comprises sorbitol, wherein the sorbitol has a concentration of 1-5 wt%.

9. The composition of claim 5, wherein the composition further comprises propylene glycol, wherein the propylene glycol has a concentration of 1-5 wt%.

10. The composition of claim 5, wherein the composition further comprises polyvinyl alcohol, wherein the polyvinyl alcohol has a concentration of 0.1-5 wt%.11 . The composition of claim 5, wherein the composition further comprises chitosan, wherein the chitosan has a concentration of 0.1-2 wt%.

12. The composition of claim 5, wherein the composition further comprises silica nanoparticles, wherein the silica nanoparticles have a concentration of 0.1-3 wt%.

13. The composition of claim 5, wherein the composition further comprises cellulose nanocrystals, wherein the cellulose nanocrystals have a concentration of 0.1-3 wt%.ATTORNEY DOCKET NO. 222204-281014. The composition of claim 5, wherein the composition further comprises ethylene vinyl acetate, wherein the ethylene vinyl acetate has a concentration of 0.1-10 wt%.

15. The composition of claim 5, wherein the composition further comprises magnesium, wherein the magnesium has a concentration of 1-50 mM.

16. The composition of claim 5, wherein the composition further comprises oils, wherein the oils have a concentration of 0.01-0.1 wt%.

17. The composition of claim 5, wherein the composition further comprises zinc, wherein the zinc has a concentration of 1-100 ppm.

18. The composition of claim 5, wherein the composition further comprises manganese, wherein the manganese has a concentration of 1-100 ppm.

19. The composition of claim 5, wherein the composition further comprises boron, wherein the boron has a concentration of 1-100 ppm.

20. The composition of claim 5, wherein the composition further comprises seaweed extract, wherein the seaweed extract has a concentration of 0.1-3 wt%.21 . The composition of claim 5, wherein the composition further comprises gelatin, wherein the gelatin has a concentration of 0.1-5 wt%.

22. The composition of claim 5, wherein the composition further comprises whey protein isolate, wherein the whey protein isolate has a concentration of 0.1-5 wt%.

23. The composition of claim 5, wherein the composition further comprises an ethylene inhibitor, wherein the ethylene inhibitor has a concentration of 1-100 ppm.

24. The composition of claim 5, wherein the composition further comprises abscisic acid, wherein the abscisic acid has a concentration of 1-500 pM.

25. The composition of claim 5, wherein the composition further comprises a jasmonic acid inhibitor, wherein the jasmonic acid inhibitor has a concentration of 10-500 pM.

26. The composition of any one of claims 1-25, wherein the composition is used to protect plants from freezing and subfreezing temperatures.

27. A method for protecting plants from freezing and subfreezing temperatures, the method comprising the steps:(a) applying a first composition on a surface of a plant via a first manner; and(b) applying a second composition on the surface of the plant via a second manner; wherein the first composition comprises: sodium alginate; and at least one polymer; wherein the second composition comprises: calcium chloride; andATTORNEY DOCKET NO. 222204-2810 wherein the first composition and the second composition combine to form a coating composite material.

28. The method of claim 27, wherein the at least one polymer is selected from polyvinyl alcohol, chitosan, polyethylene, polypropylene, polystyrene, polyacrylamide, polyacrylic acid, cellulose, starch, collagen, and gelatin.

29. The method of claim 28, wherein the at least one polymer is polyvinyl alcohol.

30. The method of claim 28, wherein the at least one polymer is chitosan.

31. The method of claim 29, wherein the polyvinyl alcohol has a concentration of 0.1-5 wt%.

32. The method of claim 30, wherein the chitosan has a concentration of 0.5-2 wt%.

33. The method of claim 27, wherein the first manner is manual application, using one or more of a brush, a roller, or a sponge.

34. The method of claim 27, wherein the first manner is application by a sprayer.

35. The method of claim 27, wherein the second manner is application by a sprayer.

36. The method of claim 27, wherein the method further comprises a step for removing the coating composite material from the plant, wherein a dissolving solution is applied to the plant.

37. The method of claim 36, wherein the dissolving solution comprises one or more of sodium citrate, sodium hexametaphosphate, ethylenediaminetetraacetic acid, sodium carbonate, and sodium bicarbonate.

38. The method of claim 27, wherein the plant is selected from cherry, peach, plum, apricot, apple, pear, and quince.

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