METHODS AND PRODUCTS FOR INHIBITING AN ETHYLENE RESPONSE

PRIORITY CLAIM

[0001] This application claims priority to Australian provisional patent application number 2014902208 filed on 10 June 2014, the content of which is hereby incorporated by reference.

FIELD

[0002] The present disclosure relates to methods and products for inhibiting an ethylene response in a plant product.

BACKGROUND

[0003] Ethylene is a molecule that has a wide range of effects on plants. It is produced by higher plants and mediates its effects through specific receptors. It acts at trace levels throughout the life of the plant by stimulating or regulating various responses, such as the ripening of fruit, the opening of flowers, and the abscission (shedding) of leaves.

[0004] Ethylene shortens the shelf life of many fruits by hastening fruit ripening and floral senescence. Ethylene also shortens the shelf life of cut flowers and potted plants by accelerating floral senescence and floral abscission.

[0005] To control the effects of ethylene on plants, a variety of methods have been developed, including inhibition of ethylene synthesis and inhibition of ethylene sensing. Inhibition of ethylene synthesis is considered less effective for reducing post harvest loss than inhibiting ethylene sensing, as the plants can no longer respond to either endogenously produced ethylene or ethylene from exogenous sources.

[0006] 1-methylcyclopropene (1-MCP) is an inhibitor of ethylene sensing and is one of the most widely used commercial agents to inhibit ripening. Typically the molecule is combined with other materials for handling and then mixed with a specific amount of water or other solution to release it into the air in enclosed indoor sites, such as coolers, truck trailers, greenhouses, storage facilities, and shipping containers.

[0007] It would be advantageous to identify other agents that have the ability to inhibit the ethylene response in plants, such as identifying agents with improved volatility and/or to provide a commercial choice. The present disclosure relates to the determination that 1- butyne is an effective inhibitor of the ethylene response in plants.

SUMMARY

[0008] The present disclosure is based on the recognition that 1-butyne is an effective inhibitor of the ethylene response in plants.

[0009] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene response in a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[0010] Certain embodiments of the present disclosure provide a method of slowing ripening of a plant product, reducing post-harvest decay of a plant product, and/or improving shelf life of a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[001 1] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene response in a plant product, the method comprising:

providing a source of 1-butyne; and

exposing a plant product to the 1-buytne from the source that has been released into the air;

thereby inhibiting the ethylene response in the plant product.

[0012] Certain embodiments of the present disclosure provide use of 1-butyne in the preparation of a formulation for inhibiting an ethylene response in a plant product.

[0013] Certain embodiments of the present disclosure provide an anti-ripening composition, the composition comprising an effective amount of 1-butyne.

[0014] Certain embodiments of the present disclosure provide a composition comprising 1- butyne and a cyclodextrin.

[0015] Certain embodiments of the present disclosure provide a composition for slowing ripening of a plant product, reducing postharvest decay of a plant product, slow softening of a plant product, inhibiting discolouration of a plant product, improving shelf life of a plant product, preventing premature wilting of a plant product, reducing leaf yellowing of a plant product, preventing premature opening of a plant product, and/or preventing premature death of a plant product, the composition comprising an effective amount of 1-butyne.

[0016] Certain embodiments of the present disclosure provide a method of inhibiting ethylene binding to an ethylene receptor in a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[0017] Certain embodiments of the present disclosure provide a post-harvest treatment product, the product comprising one or more sources of 1- butyne, wherein the one or more sources of 1-butyne provide a releasable amount of 1-butyne effective to inhibit ripening of a plant product for a given volume of air.

[0018] Certain embodiments of the present disclosure provide a plant product exposed to 1- butyne using a method and/or product as described herein

[0019] Other embodiments are disclosed herein.

BRIEF DESCRIPTION OF THE FIGURES

[0020] Exemplary embodiments will be better understood and appreciated in conjunction with the following detailed description of example embodiments taken together with the accompanying figures. It is to be understood that the following description of the figures is for the purpose of describing example embodiments only and is not intended to be limiting with respect to this disclosure. [0021] Figure 1 shows the results of pressure tests on avocados treated with 1-butyne as compared to untreated avocados over time (days).

[0022] Figure 2 shows the results of pressure tests on bananas treated with 1-butyne as compared to untreated bananas over time (days).

[0023] Figure 3 shows the results of pressure tests on persimmons treated with 1-butyne as compared to untreated bananas over time (days).

DETAILED DESCRIPTION

[0024] The present disclosure relates to methods and products for inhibiting an ethylene response in a plant product.

[0025] The present disclosure is based on the recognition that 1-butyne is an effective inhibitor of the ethylene response in plants. The efficacy of 1-butyne as an ethylene inhibiting agent has been demonstrated by controlled atmosphere trials. Exposure of different types of fruits to 1-butyne results in a significant reduction in the deterioration of the fruits by a variety of measures (as compared to untreated controls).

[0026] Certain disclosed embodiments may have one or more combinations of advantages. For example, some of the advantages of the embodiments disclosed herein include one or more of the following: identification of a molecule hereto unrecognised as being an inhibitor of an ethylene response in plants; use of a molecule with improved volatility to inhibit an ethylene response, thereby assisting in volatile penetration of the molecule; use of a molecule to inhibit an ethylene response in plants which has a lower synthesis or production cost; use of a molecule to inhibit an ethylene response in plants which permits the dose of the molecule to be increased due to its lower cost of application; to provide new products for inhibiting an ethylene response in plants; to provide one or more advantages in the art; or to provide a useful commercial choice. Other advantages of certain embodiments are disclosed herein or may be appreciated in practicing one or more embodiments. [0027] Certain embodiments of the present disclosure comprise a method of inhibiting an ethylene response in a plant product.

[0028] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene response in a plant product, the method comprising exposing the plant product to an effective amount of 1 -butyne.

[0029] The term " 1 -butyne'" as used herein refers to the alkyne with the chemical formula C ₄ H ₆ and CAS number 107-00-6, and which is sometimes referred to as ethylacetylene or ethylethyne. The molecule has the following structure:

[0030] The term "1 -butyne" includes the molecule as shown above and also any radicals or radical anions of the molecule. The molecule may be obtained commercially or may be synthesized by a method known in the art, for example as described in F. R. Morehouse and O. Maass ( 193 1 ) Canadian Journal of Research 5(3): 306-3 12.

[003 1 ] In certain embodiments, the ethylene response comprises one or more of ripening, postharvest decay, softening, discolouration, seedling triple response, plant, leaf and/or flower senescence, senescence of mature xylem cells, leaf abscission, fruit abscission, seed germination, root hair growth, epinasty, and flowering in pineapples. Other forms of ethylene responses are contemplated.

[0032] In certain embodiments, the ethylene response comprises one or more of ripening, postharvest decay, softening, and discolouration.

[0033] In certain embodiments, the plant product is a product that is susceptible to an ethylene response. In certain embodiments, the plant product is a product that is susceptible to spoilage, deterioration, decay, softening, or discolouration.

[0034] In certain embodiments, the plant product comprises a whole plant. In certain embodiments, the plant product comprises a part or portion of a plant.

[0035] In certain embodiments, the whole plant comprises one or more of an ornamental plant, a potted plant, a nursery plant, a tree, a shrub, a foliage plant, a flowering plant, a greenery plant, a field crop, a landscape plant, and an agricultural plant.

[0036] Examples of plants comprise cotton (Gossypium spp.), apples, pears, cherries (Primus avium), pecans (Carva illinoensis), grapes (Vitis vinifera), olives (e.g. Vitis vinifera and Olea europaea), coffee (Coffea arabica), snapbeans (Phaseolus vulgaris), and weeping fig (ficus benjamina), as well as dormant seedlings such as dormant seedlings of various fruit trees including apple, ornamental plants, shrubbery, and tree seedlings. In addition, shrubs which may be treated according to the present invention to inhibit an ethylene response, such as abscission of foliage, include privet (Ligustrum sp.), photinea (Photinia sp.), holly (Ilex sp.), ferns of the family Polypodiaceae, schefflera (Schefflera sp.), aglaonema (Aglaonema sp.), cotoneaster (Cotoneaster sp.), barberry (Berberis sp.), waxmyrtle (Myrica sp.), abelia (Abelia sp.), acacia (Acacia sp.) and bromeliades of the family Bromeliaceae. Other types of plants are contemplated. In certain embodiments, the plant product comprises a part or portion of the aforementioned plants.

[0037] In certain embodiments, a plant is exposed to 1-butyne as described herein to inhibit an ethylene response, such as to inhibit abscission of foliage, flowers and fruit.

[0038] In certain embodiments, an ornamental or flowering plant is exposed to 1-butyne as described herein to inhibit an ethylene response, such as senescence and/or shortening of flower life and thus prolong flower life and appearance (e.g. to delay wilting).

[0039] Examples of flowering plants comprise azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hybiscus (Hibiscus rosasanensis), snapdragons (Antirrhinum sp.), poinsettia (Euphorbia pulcherima), cactus (e.g. Cactaceae schlumbergera truncata), begonias (Begonia sp.), roses (Rosa spp.), tulips (Tulipa sp.), daffodils (Narcissus spp.), dandelions (Taraxacum ojfinale), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), lily (e.g., Lilium sp.), gladiolus (Gladiolus sp.), alstroemeria (Alstoemeria hrasiliensis), anemone (e.g., Anemone blanda), columbine (Aquilegia sp.), aralia (e.g., Aralia chinensis), aster (e.g., Aster carolinianus), bougainviilea (Bougainviilea sp.), camellia (Camellia sp.), bellflower (Campanula sp.), cockscomb (celosia sp.), falsecypress (Chamaecyparis sp.), chrysanthemum (Chrysanthemum sp.), clematis (Clematis sp.), cyclamen (Cyclamen sp.), freesia (e.g., Freesia refracta), and orchids of the family Orchidaceae. Other types of flowering plants are contemplated. Other types of plants are contemplated. In certain embodiments, the plant product comprises a part or portion of the aforementioned flowering plants.

[0040] In certain embodiments, the plant product comprises a part or portion of a plant. In certain embodiments, the plant product comprises a fruit, a vegetable, a cutting, and a cut flower. Other types of plant products are contemplated.

[0041] In certain embodiments, a fruit may be exposed to 1-butyne to inhibit an ethylene response, such as ripening.

[0042] Examples of fruits comprise tomatoes (Lycopersicon esculentum), apples (Malus domestica), bananas (Musa sapientum), pears (Pyrus communis), papaya (Carica papaya), mangoes (Mangifera indica), peaches (Prunus persica), apricots (Prunus armeniaca), nectarines (Prunus persica nectarina), oranges (Citrus sp.), lemons (Citrus limonia), limes (Citrus aurantifolia), grapefruit (Citrus paradisi), tangerines (Citrus nobilis deliciosa), kiwi (Actinidia chinenus), melons such as cantaloupe (C. cantalupensis) and musk melon (C. melo), pineapple (Aranas comosus), persimmon (Diospyros sp.), various other fruits including stone fruits, and berries such as strawberries (Fragaria), blueberries (Vaccinium sp.) and raspberries (e.g., Rubus ursinus), green beans (Phaseolus vulgaris), members of the genus Cucumis such as cucumber (C. sativus), and avocados (Persea americana). Other types of fruits are contemplated.

[0043] In certain embodiments, a vegetable may be exposed to 1-butyne as described herein to inhibit an ethylene response, such as to inhibit spoilage, ripening and/or senescence. [0044] Examples of vegetables comprise leafy green vegetables, such as lettuce (e.g., Lactuea sativa), spinach {Spinaca oleracea), and cabbage {Brassica oleracea), various roots, potatoes {Solanum tuberosum), carrots {Daucus), bulbs, onions {Allium sp.), herbs, basil {Ocimum basilicum), oregano {Origanum vulgare), dill {Anethum graveolens), soybean {Glycine max), lima beans {Phaseolus limensis), peas {Lathyrus spp ), corn {Zea mays), broccoli {Brassica oleracea italica), cauliflower {Brassica oleracea botrytis), and asparagus {Asparagus officinalis). Other types of vegetables are contemplated.

[0045] In certain embodiments, the method inhibits an ethylene response to internal ethylene. In certain embodiments, the method inhibits an ethylene response to external ethylene. In certain embodiments, the method inhibits an ethylene response to internal and external ethylene. In certain embodiments, the method inhibits an ethylene response due to endogenous ethylene. In certain embodiments, the method inhibits an ethylene response due to exogenous ethylene. In certain embodiments, the method inhibits an ethylene response due to endogenous and exogenous ethylene. In certain embodiments, the method inhibits an ethylene response due to ethylene produced from other plant products.

[0046] In certain embodiments, the method comprises gaseous exposure of 1-butyne to the plant product.

[0047] In certain embodiments, the exposing of the plant product to 1-butyne comprises gaseous exposure. In certain embodiments, the exposing of the plant product to 1-butyne comprises exposure to a vapour. In certain embodiments, the exposing of the plant product comprises exposure to a vapour comprising 1-butyne.

[0048] In certain embodiments, the exposing comprises exposure in a closed system. Examples of a closed system, or a substantially closed system, comprise a room, a controlled atmosphere room, a bag, a bin, a greenhouse, a trailer, a cold room, a cooler, a shipping container, a storage facility, and a storage container. Other types of closed systems are contemplated.

[0049] The effective amount is that amount or dose of 1-butyne that when exposed to a plant product produces the desired outcome. [0050] In certain embodiments, the concentration of 1-butyne comprises lxlO ^"1 to lxlO ^"6 gm/m ³ , lxlO ^"1 to lxlO ^"6 gm/m ³ , lxlO ^"1 to lxlO ^"5 gm/m ³ , lxlO ^"1 to lxlO ^"4 gm/m ³ , lxlO ^"1 to lxlO ^"3 gm/m ³ , lxlO ^"1 to lxlO ^"2 gm/m ³ , lxlO ^"2 to lxlO ^"6 gm/m ³ , lxlO ^"2 to lxlO ^"5 gm m ³ , 1x10 ^{" 2} to lxlO ^"4 gm/m ³ , lxlO ^"1 to lxlO ^"2 gm/m ³ , lxlO ^"3 to lxlO ^"6 gm/m ³ , lxlO ^"3 to lxlO ^"5 gm m ³ , lxlO ^"3 to lxlO ^"4 gm/m ³ , lxlO ^"4 to lxlO ^"6 gm/m ³ , lxlO ^"4 to lxlO ^"5 gm/m ³ or lxlO ^"5 to lxlO ^"6 gm/m ³ .

[0051] In certain embodiments, the exposing comprises gaseous exposure in a closed system.

[0052] In certain embodiments, the exposing comprises gaseous exposure to a concentration of 1-butyne at a concentration in the air/atmo sphere as described herein.

[0053] In certain embodiments, the concentration of 1-butyne in the atmosphere comprises lxlO ^"1 to lxlO ^"6 gm/m ³ .

[0054] In certain embodiments, the concentration of 1-butyne in the atmosphere is lxlO ^"1 to lxlO ^"6 gm/m ³ , lxlO ^"1 to lxlO ^"6 gm/m ³ , lxlO ^"1 to lxlO ^"5 gm/m ³ , lxlO ^"1 to lxlO ^"4 gm m ³ , 1x10 ^{" 1} to lxlO ^"3 gm/m ³ , lxlO ^"1 to lxlO ^"2 gm/m ³ , lxlO ^"2 to lxlO ^"6 gm/m ³ , lxlO ^"2 to lxlO ^"5 gm/m ³ , lxlO ^"2 to lxlO ^"4 gm m ³ , lxlO ^"1 to lxlO ^"2 gm/m ³ , lxlO ^"3 to lxlO ^"6 gm m ³ , lxlO ^"3 to lxlO ^"5 gm/m ³ , lxlO ^"3 to lxlO ^"4 gm m ³ , lxlO ^"4 to lxlO ^"6 gm/m ³ , lxlO ^"4 to lxlO ^"5 gm/m ³ or lxlO ^"5 to lxlO ^"6 gm m ³ .

[0055] In certain embodiments, the exposure of the plant product to 1-butyne comprises post-harvest exposure to the 1-butyne.

[0056] In certain embodiments, the exposure of the plant product to 1-butyne comprises pre-harvest exposure to the 1-butyne.

[0057] In certain embodiments, the exposure of the plant product to 1-butyne comprises pre-harvest and post-harvest exposure to the 1-butyne.

[0058] In certain embodiments, the exposure to 1-butyne comprises a temperature of exposure of 30°C or less, 25°C or less, 20°C or less, 15°C or less, 10°C or less, 5°C or less, 4°C or less, 3°C or less, 2°C or less, or 1°C or less. In certain embodiments, the temperature is in the range from -2°C to 30°C, 0°C to 30°C, 1°C to 30°C, 2°C to 30°C, 3°C to 30°C, 4°C to 30°C, 5°C to 30°C, 10°C to 30°C, 15°C to 30°C, 20°C to 30°C, 25°C to 30°C, -2°C to 25°C, 0°C to 25°C, 1°C to 25°C, 2°C to 25°C, 3°C to 25°C, 4°C to 25°C, 5°C to 25°C, 10°C to 25°C, 15°C to 25°C, 20°C to 25°C, -2°C to 20°C, 0°C to 20°C, 1°C to 20°C, 2°C to 20°C, 3°C to 20°C, 4°C to 20°C, 5°C to 20°C, 10°C to 20°C, 15°C to 20°C, -2°C to 15°C, 0°C to 15°C, 1°C to 15°C, 2°C to 15°C, 3°C to 15°C, 4°C to 15°C, 5°C to 15°C, 10°C to 15°C, -2°C to 10°C, 0°C to 10°C, 1°C to 10°C, 2°C to 10°C, 3°C to 10°C, 4°C to 10°C, 5°C to 10°C, -2°C to 5°C, 0°C to 5°C, 1°C to 5°C, 2°C to 5°C, 3°C to 5°C, or 4°C to 5°C.

[0059] In certain embodiments, the exposure comprises exposing the plant product to 1- butyne for a time of at least 6 hours, at least 12 hours, at least 18 hours, or at least 24 hours. In certain embodiments, the exposure comprises exposing the plant product to 1-butyne for a time of 6 hours or greater, 12 hours or greater, 18 hours or greater, or 24 hours or greater. In certain embodiments, the exposure comprises exposing the plant product to 1-butyne for a time of 6 to 36 hours, 6 to 24 hours, 6 to 18 hours, 6 to 12 hours, 12 to 36 hours, 12 to 24 hours, 12 to 18 hours, 18 to 36 hours, 18 to 24 hours or 24 to 36 hours. Other lengths of time are contemplated.

[0060] In certain embodiments, the aforementioned times of exposure comprise exposure at a temperature as described herein, such as at room temperature or at a colder temperature.

[0061] In certain embodiments, the aforementioned times of exposure comprises exposure at a temperature in the range from -2°C to 30°C.

[0062] In certain embodiments, the aforementioned times of exposure comprise exposure at a temperature of 30°C or less, 25°C or less, 20°C or less, 15°C or less, 10°C or less, 5°C or less, 4°C or less, 3°C or less, 2°C or less, or 1°C or less. In certain embodiments, the time of exposure comprises exposure at a temperature in the range from -2°C to 30°C, 0°C to 30°C, 1°C to 30°C, 2°C to 30°C, 3°C to 30°C, 4°C to 30°C, 5°C to 30°C, 10°C to 30°C, 15°C to 30°C, 20°C to 30°C, 25°C to 30°C, -2°C to 25°C, 0°C to 25°C, 1°C to 25°C, 2°C to 25°C, 3°C to 25°C, 4°C to 25°C, 5°C to 25°C, 10°C to 25°C, 15°C to 25°C, 20°C to 25°C, -2°C to 20°C, 0°C to 20°C, 1°C to 20°C, 2°C to 20°C, 3°C to 20°C, 4°C to 20°C, 5°C to 20°C, 10°C to 20°C, 15°C to 20°C, -2°C to 15°C, 0°C to 15°C, 1°C to 15°C, 2°C to 15°C, 3°C to 15°C, 4°C to 15°C, 5°C to 15°C, 10°C to 15°C, -2°C to 10°C, 0°C to 10°C, 1°C to 10°C, 2°C to 10°C, 3°C to 10°C, 4°C to 10°C, 5°C to 10°C, -2°C to 5°C, 0°C to 5°C, 1°C to 5°C, 2°C to 5°C, 3°C to 5°C, or 4°C to 5°C.

[0063] In certain embodiments, the plant product is exposed to the 1-butyne within 5 days of harvest, within 4 days of harvest, within 3 days of harvest, within 2 days of harvest, within 1 day of harvest, within 18 hours of harvest, within 12 hours of harvest, within 6 hours of harvest, within 5 hours of harvest, within 4 hours of harvest, within 3 hours of harvest, within 2 hours of harvest, within 1 hour of harvest or immediately following harvest. In certain embodiments, the plant product is exposed to the 1-butyne immediately following harvest.

[0064] In certain embodiments, the method comprises providing a source of 1-butyne. In certain embodiments the method comprises providing a source of 1-butyne in a gaseous form, a liquid form or a gaseous/liquid form. In certain embodiments the method comprises providing a sealed source of 1-butyne. In certain embodiments the method comprises providing a sealed source of 1-butyne in a gaseous form, a liquid form or a gaseous/liquid form. In certain embodiments, the 1-butyne is in a complexed form. In certain embodiments, the 1-butyne is in a chelated form. In certain embodiments, the 1-butyne is incorporated in a solid matrix. Other forms are contemplated.

[0065] In certain embodiments, the method comprises providing a sealed source of 1-butyne and releasing the 1-butyne from the source.

[0066] In certain embodiments, the method comprises providing a sealed source of 1-butyne in a gaseous form, a liquid form or a gaseous/liquid form and releasing gaseous 1-butyne from the sealed source. Other forms of 1-butyne are contemplated.

[0067] Various methods are known in the art for detecting and/or measuring 1-buytne, including for example gas chromatography, kitagawa tubes, chemical sensors or other types of plants products that are sensitive to ethylene. [0068] In certain embodiments, the exposing comprises gaseous exposure in a closed system. Examples of closed systems are as described herein.

[0069] In certain embodiments, the exposing comprises contacting the plant product with 1- butyne.

[0070] In certain embodiments, the exposing comprising exposing the plant product to a composition comprising 1-butyne. Compositions comprising 1-butyne are as described herein. In certain embodiments, the exposing comprises contacting the plant product with a composition comprising 1-butyne. In certain embodiments, the exposing comprises gaseous exposure of the plant product to 1-butyne released from a composition.

[0071] In certain embodiments, the exposing comprises spraying and/or dipping.

[0072] For example, the exposing may comprise exposure as a liquid, exposure as a semi- liquid, exposure as a semi-solid, exposure as a gel, or exposure as a solid. In certain embodiments, the exposing comprises exposing the plant to a liquid composition, to a semi- liquid composition, to a semi-solid composition, to a gel, or to a solid composition.

[0073] In certain embodiments, the exposing occurs at a temperature in the range from -2°C to 30°C. Other temperatures, and ranges of temperatures, are as described herein.

[0074] In certain embodiments, the 1-butyne is complexed with an agent. In certain embodiments, the 1-butyne is chelated with an agent.

[0075] In certain embodiments, the agent comprises an encapsulating agent. In certain embodiments, the encapsulating agent comprises a cyclodextrin. Examples of cyclodextrins include a-cyclodextrin, β-cyclodextrin and/or γ-cyclodextrin. In certain embodiments, the encapsulating agent comprises α-cyclodextrin. Methods for use of cyclodextrin to encapsulate agents ae known in the art, for example as described in US Patent 8,414,989.

[0076] In certain embodiments, the method comprises providing a source of 1-butyne. In certain embodiments, the source of 1-butyne comprises 1-butyne alone, a solution comprising 1-butyne and/or a composition comprising 1-butyne. [0077] In certain embodiments, the method comprises providing a source of 1-butyne and exposing the plants to the 1-butyne from the source.

[0078] In certain embodiments, the method comprises providing a source of 1-butyne and exposing the plants to the 1-butyne from that source that has been released into the air.

[0079] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene response in a plant product, the method comprising:

providing a source of 1-butyne; and

exposing a plant product to the 1-buytne from the source that has been released into the air;

thereby inhibiting the ethylene response in the plant product.

[0080] In certain embodiments, the methods of the present disclosure may be used to slow ripening, reduce postharvest decay, slow softening, inhibit discolouration, improve shelf life, prevent premature wilting, reduce leaf yellowing, to prevent premature opening, and/or to prevent premature death. Other uses are contemplated.

[0081] Certain embodiments of the present disclosure provide a method of slowing ripening of a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[0082] Certain embodiments of the present disclosure provide a method of reducing postharvest decay of a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[0083] Certain embodiments of the present disclosure provide a method of improving shelf life of a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[0084] Certain embodiments of the present disclosure provide a method of slowing ripening of a plant product, reducing post-harvest decay of a plant product, and/or improving shelf life of a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne. [0085] Certain embodiments of the present disclosure provide a plant product exposed to 1- butyne according to a method as described herein. Examples of plants products are as described herein.

[0086] Certain embodiments of the present disclosure provide use of 1-butyne in the preparation of a formulation or composition for inhibiting an ethylene response in a plant product. Formulations and compositions comprising 1-butyne are as described herein.

[0087] Certain embodiments of the present disclosure provide a composition for inhibiting an ethylene response in a plant product, the composition comprising an effective amount of 1-butyne.

[0088] Certain embodiments of the present disclosure provide a composition for slowing ripening of a plant product, reducing postharvest decay of a plant product, slow softening of a plant product, inhibiting discolouration of a plant product, improving shelf life of a plant product, preventing premature wilting of a plant product, reducing leaf yellowing of a plant product, preventing premature opening of a plant product, and/or preventing premature death of a plant product, the composition comprising 1-butyne.

[0089] The compositions may be applied in gaseous, liquid or solid form or a mixture of any combination thereof, by contacting the composition with the plant product to be treated. Other forms are contemplated. Such compositions may include an inert carrier.

[0090] In certain embodiments, the composition is a gaseous composition, a liquid composition, an aerosol composition, a semi-solid composition or a solid composition. Other types of compositions are contemplated.

[0091] For example, a gaseous composition may comprise one or more gases and 1-butyne.

[0092] A liquid composition may comprise one or more organic solvents and 1-butyne. Examples of organic solvents are hydrocarbons such as hexane, benzene, toluene, xylene, kerosene, diesel oil, fuel oil and petroleum naphtha, ketones such as acetone, methyl ethyl ketone and cyclohexanone, chlorinated hydrocarbons such as carbon tetrachloride, esters such as ethyl acetate, amyl acetate and butyl acetate, ethers, e.g., ethylene glycol monomethyl ether and diethylene glycol monomethyl ether, alcohols, e.g., ethanol, methanol, isopropanol, amyl alcohol, ethylene glycol, propylene glycol, butyl carbitol acetate and glycerine.

[0093] Mixtures of water and/or organic solvents, either as solutions or emulsions, can also be employed.

[0094] A solid composition may comprise 1-butyne and a complexing agent. In certain embodiments, the complexing agent comprises an encapsulating agent, such as a cyclodextrin. Examples of cyclodextrins include a cyclodextrin, β cyclodextrin and γ cyclodextrin.

[0095] Solid and liquid compositions may comprise one or more adjuvants, carriers, extenders, binders, lubricants, surfactants and/or dispersants, wetting agents, spreading agents, dispersing agents, stickers, adhesives, defoamers, thickeners, and emulsifying agents. Such adjuvants commonly used in the art can be found in the John W. McCutcheon, Inc. publication Detergents and Emulsifiers, Annual, Allured Publishing Company, Ridgewood, N.J., U.S.A.

[0096] Solid, liquid, and gaseous formulations can be prepared by various conventional procedures.

[0097] In certain embodiments, a composition comprising 1-butyne is used to slow ripening in plants products.

[0098] Certain embodiments of the present disclosure provide an anti-ripening composition, the composition comprising an effective amount of 1-butyne.

[0099] In certain embodiments, the composition comprises a solvent, such as an organic solvent. Organic solvents are as described herein. In certain embodiments, the composition comprises water as a solvent. For example, 1-butyne may be provided in conjunction with an encapsulating agent, and the complex mixed in water.

[00100] In certain embodiments, the composition comprises a complexing agent. In certain embodiments, the complexing agent is a cyclodextrin based encapsulating agent. For example, cyclodextrin molecules can be used to produce a commercially stable source of the 1-butyne. For example, 1-butyne may be formulated with , β or γ cyclodextrin at a ratio of 3 to 5%. Such compositions may further comprise a release agent (eg acid, carbonate, bicarbonate) that assist with dissipation of the encapsulated 1-butyne when placed in water. Application of water or a buffer solution can then be used to initiate release of the 1-butyne from its cyclodextrin matrix.

[00101] Certain embodiments of the present disclosure provide a composition comprising 1-butyne and a complexing agent. Certain embodiments of the present disclosure provide a composition comprising 1-butyne and a chelating agent. Certain embodiments of the present disclosure provide a composition comprising 1-butyne and an encapsulating agent. In certain embodiments, the complexing agent comprises a cyclodextrin.

[00102] Certain embodiments of the present disclosure provide a composition comprising 1-butyne and a cyclodextrin. Cyclodextrins are as described herein.

[00103] Certain embodiments of the present disclosure provide an anti-ripening product, the product comprising the following components:

1-butyne; and

a solvent for 1-butyne,

wherein the components are provided in a form for mixing of the components to produce a volatile solution of 1-butyne for exposure to a plant product. Solvents are as described herein.

[00104] Certain embodiments of the present disclosure provide a package comprising a composition as described herein, and/or one or more delivery vehicles.

[00105] Certain embodiments of the present disclosure provide a post-harvest treatment product.

[00106] Certain embodiments of the present disclosure provide a post-harvest treatment product, the product comprising one or more sources of 1- butyne, wherein the one or more sources of 1-butyne provide a releasable amount of 1-butyne effective to inhibit ripening of a plant product for a given volume of air.

[00107] In certain embodiments, the one or more sources comprise a plurality of sources. Sources of 1-butyne are as described herein. Methods for exposing a plant product to 1- butyne are as described herein. [00108] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene response in a plant product in a container, the method comprising using a post- harvest product as described herein to release 1- butyne from one or more sources of 1- butyne sufficient to provide an amount of 1-butyne in the container to inhibit an ethylene response in the plant product in the container.

[00109] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene response in a plant product, the method comprising exposing the plant product to a formulation, composition and/or a product as described herein.

[001 10] Certain embodiments of the present disclosure provide a method of inhibiting an ethylene receptor in a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[001 1 1] Certain embodiments of the present disclosure provide a method of inhibiting ethylene binding to an ethylene receptor in a plant product, the method comprising exposing the plant product to an effective amount of 1-butyne.

[00112] Methods for exposing a plant product to 1-butyne are as described herein.

[00113] Methods for determining the activity of an ethylene receptor are known in the art, for example as described in Biology and Biotechnology of the Plant Hormone Ethylene III (2002) edited by Miguel Vendrell, Harry Klee, Jean Claude Pech & Felix Romojaro, IOS press. In certain embodiments, an indirect assay for ethylene receptor activity may be used, such as ripening.

[001 14] Certain exemplary embodiments are illustrated by some of the following examples. It is to be understood that the following description is for the purpose of describing particular embodiments only and is not intended to be limiting with respect to the above description.

EXAMPLE 1 - 1-utyne functions as a post harvest ethylene inhibiting agent for fresh agricultural produce

[001 15] The production of 1-butyne was achieved by conventional organic synthesis, and was provided by Sigma- Aldrich of 3050 Spruce Street, Saint Louis MO 63103, USA. Various methods are known in the art to synthesize 1 -butyne, for example using acetylene.

[001 16] Owing to its particularly low boiling point of approximately 8 degrees Centigrade, a means of long term storage for application was achieved by containment in high pressure stainless steel tubes with gas tight end closures. No degradation or deterioration of the product was observed even after 1 year of storage at ambient temperatures and confirmation of this was achieved by in house GC/MS analysis, demonstrating that 1 -butyne is stable over extended periods of time and that the containment means was also suitable for the long term storage of 1 -butyne.

[001 17] An initial trial conducted involved the treatment of a 500 m ³ controlled atmosphere room containing 2 bins of fresh post harvest Golden delicious apples which was achieved by opening a sealed metal tube containing no more than 0.5 ml (0.339 gm) of 1- butyne in the controlled atmosphere room and leaving the "cracked open" tube in a well ventilated location in the controlled atmosphere room for thorough distribution for 24 hours, resulting in a vapour concentration of 0.000678 grams per m ³ throughout the controlled atmosphere room. Whilst this approaches the critical concentration required for effective ethylene inhibition of this variety of apple (based on applications of 1-MCP) it was also found that a double dose was more effective and provided an assurance buffer for efficacy and obviated the need for stringent sealing of controlled atmosphere rooms.

[001 18] It was found that for the apples there was a significant difference in the ripening and storage of the apples when exposed to 1 -butyne, as compared to the untreated controls.

[001 19] Discussion

[00120] The mode of action of 1 -butyne reduces the undesirable production of ethylene during the ripening process of fruit, which otherwise causes degradation. The historical use of 1-MCP, which appears to operate in an identical mode to 1 -butyne in relation to the biochemistry of ethylene inhibition, has adequately demonstrated the nutritional quality of stored treated fruit is no different to that of untreated fresh fruit and no residual trace of the treatment (or any by-product of it) can be detected by analytical instruments. [00121] Efficacy

[00122] Owing to the lower cost of production/application of 1-butyne in comparison to 1-MCP, the treatment dosage can be increased without any detriment to ensure against possible loss from poor sealing of controlled atmosphere rooms. As a more volatile compound than 1-MCP, the distribution of the vapour in a controlled atmosphere room is very rapid, as is the reaction kinetics of its ethylene inhibition.

EXAMPLE 2 - Efficacy of 1-butyne

[00123] The efficacy of 1-butyne as an ethylene inhibiting agent has been established by controlled testing conducted in an Adelaide Hills orchard. The following tests confirmed that 1-butyne is efficacious with regards to the ethylene blocking mechanism in fruit.

[00124] A small quantity of 1-butyne was synthesised for trials with a variety of fruit that spoils rapidly in order to determine efficacy. Confirmation of the molecular composition of 1- butyne was done initially with in-house analytical instrumentation, and also by a NATA registered ALS laboratory confirming the presence of 1-butyne in the synthesised material.

[00125] Bananas, avocados and persimmons were selected, however apples were also included to gain more quantitative data from specific testing normally associated with apples, and being the major product from the orchard, a readily available test subject.

[00126] The test apparatus consisted of 2 empty wooden fruit bins with (new) clear plastic 'pallet bags' placed over each to retain air composition within the given volume. The pallet bags were taped to the floor of the controlled atmosphere room in order to create an effective seal and each bin marked accordingly. Prior to sealing the bins, standard 'market size boxes' (-12-15 kg) of fruit, being bananas, avocados, persimmons and apples were placed in each bin. The bins remained in the controlled atmosphere room which had not ever been treated with 1-MCP and held at 1°C over the duration of the test.

[00127] It should be noted that the bananas were sourced green and first treated with 1- butyne in the above test and subsequently gassed with ethylene as this is standard practice for accelerated ripening. Despite this, the 1-butyne treated fruit still maintained integrity in comparison to the control. [00128] The marked bins containing the boxes of fruit remained in the controlled atmosphere room, for 6 hours to lower the temperature of the fruit. The control bin was placed adjacent to the test bin which received a precise charge of 1-butyne administered from a glass syringe micro pipette capable of delivering 1.0 uL. A 1.0 uL volume of 1-butyne was injected through a 'pin prick' hole in the plastic pallet bag, and sealed with tape. This represents an equivalent dose for a 500 m ³ controlled atmosphere room of only 0.5 ml of 1- butyne.

[00129] The control and test bins were left undisturbed for 24 hours and then opened to retrieve the boxes of fruit, where they were stored at ambient temperature thereafter. This fluctuated from 6°C at night to ~28°C during the day and represents typical ambient storage conditions for purchased fruit.

[00130] A series of quantitative tests were conducted on both the control and 1-butyne treated fruit for a period of up to 2 weeks, at which time most of the control fruit had deteriorated significantly. In the case of apples, which showed the least degradation in the control, tests were conducted after 2 months and significant differences then became quite apparent in the treated and control.

[00131] The quantitative tests applied were:

• Pressure test - kg/cm (also referred to as firmness test)

• Colour index - numeric index from colour chart comparator

• Starch index - chart comparator showing ranges of % sugar conversion from cross sections of fruit after addition of 1% Iodine

[00132] The results of pressure testing are shown in Figure 1 (avocado), Figure 2 (banana), and Figure 3 (persimmon). As can be seen, all three fruits showed significantly improved firmness as compared to control over the entire test period, demonstrating the efficacy of 1-butyne to inhibit ripening.

[00133] Additional qualitative testing was also performed on other fruit. During the cherry season, 1-butyne was applied to a standard box of 'Dame Roma' and 'Kordia' variety cherries. A similar application of 1-butyne was applied as described above and a control of untreated cherries was used to make a comparison.

[00134] The boxes of treated and control cherries were left at 1°C for 24 hours after which time both lots were held at ambient temperature. Careful observation was made of both lots and no apparent difference could be seen in the first 2 days, however on day 3 the treated boxes of cherries had a brighter and fresher appearance than the control cherries. A dramatic difference became evident after day 4 as the stalks of both varieties of the control cherries had started to brown significantly and the fruit was on the verge of being unsaleable. Even after 2 weeks, by which time the control cherries were virtually rotten, the 1-butyne treated cherry varieties remained in saleable condition with green stalks and plump skins. They were subsequently consumed and reported to have tasted as though they had just been picked.

[00135] A further test was conducted with fresh table grapes and results indicate that the abscission mechanism is blocked by similar applications of 1-butyne. Virtually all grapes remained on their stalk after treatment with 1-butyne as opposed to the control which lost up to half their berries at ambient temperature.

[00136] Additional testing of a variety of plum (Tegan Blue) was performed with outstanding results given the treated fruit stored in controlled atmosphere conditions for 3 months displayed qualities to that of fruit having just been picked, which was considerable remarkable for plums.

EXAMPLE 3 - 1 -butyne containing products to slow ripening

[00137] A variety of different products containing 1-butyne are envisaged for improving the quality of plant products when stored. For example, products for use in storage containers or for use in storage bags are described below.

[00138] (i) Product for use in storage containers, such as controlled atmosphere rooms.

[00139] A product may be provided that contains 1-butyne, for example in metal containers with a breakable seal, so that the amount of 1-butyne is sufficient to prevent ripening for a specific volume of atmosphere. For example, for small size storage containers

(eg 10m 3 ^J - 30m 3 ^J ) one product may contain 3 metal containers with 1-butyne whereby each of the containers contains sufficient releasable 1-butyne (eg 10 μΐ) for each 10 m ³ of atmosphere.

3 3

[00140] A product for intermediate size storage containers (eg 125 m -500 m ) may use 5 containers 1-butyne, each container having sufficient releasable 1-butyne (eg 100 μΐ) for each 100 m ³ of atmosphere.

[00141] The 1-butyne container may hold, for example, 1-butyne alone or utilise 1-butyne in a solvent or in a sold matrix.

[00142] (ii) Product for use in storage bags.

[00143] A product may be provided that contains 1-butyne, for example in one or more small containers, sachets or other form with an openable seal, so that the amount of 1-butyne is sufficient to prevent ripening of a plant product in a storage bag, such as used for bananas. For example, a product may be supplied that has separate containers having around 1 μg of 1- buytne in a releasable form for use in storage bags.

[00144] Although the present disclosure has been described with reference to particular examples, it will be appreciated by those skilled in the art that the disclosure may be embodied in many other forms.

[00145] Reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that this prior art forms part of the common general knowledge in any country.

[00146] As used herein, the singular forms "a," "an," and "the" may refer to plural articles (i.e., "one or more," "at least one," etc.) unless specifically stated otherwise.

[00147] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.

[00148] The subject headings used herein are included only for the ease of reference of the reader and should not be used to limit the subject matter found throughout the disclosure or the claims. The subject headings should not be used in construing the scope of the claims or the claim limitations.

[00149] Future patent applications may be filed on the basis of the present application, for example by claiming priority from the present application, by claiming a divisional status and/or by claiming a continuation status. It is to be understood that the following claims are provided by way of example only, and are not intended to limit the scope of what may be claimed in any such future application. Nor should the claims be considered to limit the understanding of (or exclude other understandings of) the present disclosure. Features may be added to or omitted from the example claims at a later date.