A method to delay plant senescence and restore abscisic acid-inhibited seed germination

Abstract

This invention relates to a method for delaying plant senescence and restoring abscisic acid-inhibited seed germination. Specifically, this invention provides the use of oxaloyl for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination; or for preparing a composition or formulation for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination. This invention also discovers for the first time that oxaloyl can also be used for (a) delaying plant hormone-induced senescence; (b) restoring the phenotype of ABA-inhibited seed germination; and (c) preserving plants.

Description

Technical Field

[0001] This invention relates to the field of agronomy, and more specifically, to a method for delaying plant senescence and restoring seed germination inhibited by abscisic acid. Background Technology

[0002] Plant senescence is a crucial stage in the growth and development of plants, reflecting their adaptive response to the environment. As sessile organisms, plants cannot escape danger like animals. Therefore, through long-term biological evolution, plants have evolved senescence mechanisms to adapt to adverse growing environments, prematurely ending their life cycle and producing offspring. Research categorizes plant senescence into four main types: partial senescence, aboveground tissue senescence, synchronous senescence, and whole-plant senescence. Annual plants, such as rice, wheat, and Arabidopsis, flower and fruit only once in their lifetime. They die after seed maturation, so whole-plant senescence promotes premature seed maturation, ending the plant's life cycle. However, plants like bananas and gladioli exhibit aboveground senescence. They maintain the life of their underground tissues, such as roots and tubers, while the aboveground tissues age and die after fruiting. The following year, the living underground parts will nurture the next generation of leaves, flowers, and fruits. There are many examples of partially aging and dying plants. These plants, like eucalyptus and pine, retain the vitality of their stem and root tips, even as most of their tissues age, and can still differentiate new branches and leaves. Trees like elm and maple, on the other hand, exhibit aging and regeneration that change with the seasons—falling in autumn and sprouting in spring, a cyclical process of life. Scientists define this type of aging as synchronous aging.

[0003] The aging process of plants is accompanied by important structural and physiological and biochemical changes. For example, the rate of cell division slows down, cell size decreases, cell membrane fluidity changes, intercellular connectivity decreases, photosynthesis decreases, starch content decreases, chlorophyll decreases while anthocyanin accumulation increases, protein degradation accelerates, protein synthesis rate decreases, and total RNA volume decreases, etc.

[0004] Seed germination, as a fundamental life process, has an efficiency that significantly impacts crop yield. Effective seed germination can significantly increase crop output. However, during seed storage and transportation, germination efficiency and yield can decrease. Current research indicates that a major limiting factor for seed germination is the influence of plant hormones, such as ABA levels. During germination, seeds first rapidly absorb water from the soil, and some secrete a mucus that softens the seed coat. This step is considered a critical rate-limiting step in seed germination because the seed needs to activate various physiological and biochemical regulatory factors, such as relevant enzymes, to secrete the mucus that softens the hard seed coat. After the seed coat softens, the radicle begins to develop. At this stage, the seed actively absorbs water and nutrients from the soil through the formation of the radicle to prepare for the development of the above-ground parts, such as stems and leaves. The final step in seed germination involves the comprehensive activation, integration, and redistribution of the metabolic levels of the seedling embryo, promoting the overall growth of the entire seedling.

[0005] Besides their own sugar stores, seeds require specific environmental conditions to germinate. Water is an essential element for seed germination. Some overly dry seeds need to absorb sufficient water to promote germination. Water helps soften the seed coat, converting insoluble substances into soluble substances to provide nutrients for germination. Oxygen is another necessary element for seed germination. It provides energy for the seed to activate its metabolism in the early stages, aiding in aerobic respiration before green leaves emerge. Appropriate temperature is also crucial; generally, seed germination occurs within the range of 25-30℃. However, the optimal temperature varies significantly among different seeds. Some seeds can germinate at temperatures fluctuating between 5-40℃. Light is also a vital factor in seed germination; many seeds require sunlight to germinate.

[0006] Currently, there are very limited reports on soil bacteria improving seed germination efficiency and inhibiting senescence.

[0007] Therefore, there is an urgent need in this field to develop a new method that can effectively improve the seed germination efficiency inhibited by ABA and significantly delay the senescence of plant leaves. Summary of the Invention

[0008] The purpose of this invention is to provide a new method that can effectively improve the ABA-inhibited seed germination efficiency and significantly delay the senescence of plant leaves.

[0009] A first aspect of the invention provides the use of diacetyl for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination; or for preparing a composition or formulation for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination.

[0010] In another preferred embodiment, the formulation or composition is also used for one or more purposes selected from the group consisting of:

[0011] (a) Delaying plant hormone-induced senescence;

[0012] (b) Restoring the phenotype of ABA-inhibited seed germination;

[0013] (c) Preserving plants (such as leafy green plants);

[0014] (d) Maintain good germination efficiency for seeds during long-term transport and storage.

[0015] In another preferred embodiment, the plant hormone is selected from the group consisting of ABA, jasmonic acid, salicylic acid, or combinations thereof.

[0016] In another preferred embodiment, the diacetyl is derived from rhizosphere growth-promoting bacteria, such as Bacillus amyloliquefaciens GB03; lactic acid bacteria, such as Lactobacillus harbinensis; or cocoa beans.

[0017] In another preferred embodiment, the composition is an agricultural composition.

[0018] In another preferred embodiment, the composition comprises (a) diacetyl; and (b) an agronomically acceptable carrier.

[0019] In another preferred embodiment, the composition further includes other substances for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination.

[0020] In another preferred embodiment, the other substances used to (a) delay plant senescence and / or (b) promote ABA-inhibited seed germination include: diacetyl small molecules, diacetyl salts, BA (6-benzyladenine), vitamin E, gibberellin, and auxin.

[0021] In another preferred embodiment, the dosage form of the composition or preparation is selected from the group consisting of solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols, or combinations thereof.

[0022] In another preferred embodiment, the plants include those that require long-distance transportation or require storage in the dark and / or in low light conditions.

[0023] In another preferred embodiment, the plant comprises an detached plant.

[0024] In another preferred embodiment, the plants include agricultural plants, horticultural plants, and forestry plants.

[0025] In another preferred embodiment, the plants include woody plants and herbaceous plants.

[0026] In another preferred embodiment, the plant includes a whole plant, organs (such as roots, stems, leaves, branches, flowers, fruits, seeds), tissues (such as callus), or cells.

[0027] In another preferred embodiment, the plants include those belonging to the Solanaceae, Fabaceae, Musaceae, Bananaaceae, Oliveaceae, Brassicaceae, Chenopodiaceae, Liliaceae, and Poaceae families.

[0028] In another preferred embodiment, the plants include plants from the genera *Solanum*, *Soybean*, *Tobacco*, *Arabidopsis*, *Brassica*, *Allium*, *Spinach*, and *Oryza*.

[0029] In another preferred embodiment, the plants include Arabidopsis thaliana, purslane, potato, spinach, rice, leek, bermudagrass, and spotted bamboo.

[0030] A second aspect of the present invention provides a composition comprising:

[0031] (i) Diacetyl;

[0032] (ii) Other substances used to (a) delay plant senescence; and / or (b) promote ABA-inhibited seed germination;

[0033] (iii) An agriculturally acceptable carrier.

[0034] In another preferred embodiment, the composition comprises an agricultural composition.

[0035] In another preferred embodiment, the composition includes a preservative and a germination promoter.

[0036] In another preferred embodiment, the dosage form of the composition is selected from the group consisting of solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols, or combinations thereof.

[0037] In another preferred embodiment, the composition contains 0.0001-99 wt%, preferably 0.1-90 wt%, of component (a) based on the total weight of the composition.

[0038] In another preferred embodiment, the composition contains 0.0001-99 wt%, preferably 0.1-90 wt%, of component (b) based on the total weight of the composition.

[0039] In another preferred embodiment, the content of component (a) in the agricultural composition is 0.01-400 micromoles, more preferably 0.02-60 micromoles, even more preferably 0.02-20 micromoles, and even more preferably 0.264-20 micromoles.

[0040] In another preferred embodiment, the concentration of component (a) in the agricultural composition is 5-400 μM, more preferably 20-300 μM, and even more preferably 50-200 μM.

[0041] In another preferred embodiment, the weight ratio of component (a) to component (b) is 100:1-0.01:1, more preferably 10:1-0.1:1, and even more preferably 2:1-0.5:1.

[0042] In another preferred embodiment, the components (a) and (b) constitute 0.01-99.99 wt% of the total weight of the composition, more preferably 0.1-90 wt%, and even more preferably 1-80 wt%.

[0043] In another preferred embodiment, the other substances used to (a) delay plant senescence and / or (b) promote ABA-inhibited seed germination include: diacetyl small molecules, diacetyl salts, BA (6-benzyladenine), vitamin E, gibberellin, and auxin.

[0044] A third aspect of the present invention provides the use of the composition described in the second aspect of the present invention for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination.

[0045] A fourth aspect of the present invention provides a method for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination, comprising the steps of:

[0046] The plant may be treated with diacetyl or with the composition described in the second aspect of the invention.

[0047] In another preferred embodiment, the method is performed under conditions of light avoidance and / or low light.

[0048] In another preferred embodiment, the weak light refers to light intensity ≤ 60 micromoles / m²·second, more preferably 0-60 micromoles / m²·second, and even more preferably 0-30 micromoles / m²·second.

[0049] In another preferred embodiment, the application is selected from the group consisting of spraying, watering, drip irrigation, misting, coating, injection, or other methods known to those skilled in the art.

[0050] In another preferred embodiment, the application may be a single application, repeated application, or continuous application.

[0051] In another preferred embodiment, the application method is to apply it to plants, or to the soil surrounding the plants, or to detached leafy green vegetables.

[0052] In another preferred embodiment, the applied dose is 0.01-400 μmol / plant, more preferably 0.02-60 μmol / plant, even more preferably 0.02-20 μmol / plant, and still more preferably 0.066-20 μmol / plant.

[0053] In another preferred embodiment, the applied dose is 5-400 μM / plant, more preferably 20-300 μM / plant, and even more preferably 50-200 μM / plant.

[0054] The fifth aspect of the present invention provides a method for preserving plants, the method comprising applying diacetyl or the composition described in the second aspect of the present invention to the plants.

[0055] In another preferred embodiment, the method is performed under conditions of light avoidance and / or low light.

[0056] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0057] The following figures are used to illustrate specific embodiments of the present invention and are not intended to limit the scope of the invention as defined by the claims.

[0058] Figure 1 The results show the phenotypes of various small-molecule volatiles, including diacetyl, on the anti-aging effects of detached leaf tissue.

[0059] Figure 2 [A] Phenotype showing that diacetyl effectively delays the senescence of the entire plant. [B] Representative single plant senescence phenotypes under diacetyl-treated and untreated conditions.

[0060] Figure 3 [A] Diacetyl delays ABA-induced plant senescence. [B] Total chlorophyll content in plants; error bars represent 6 biological replicates, n>18. [C] Quantitative analysis of ion osmotic efficiency; error bars represent 6 biological replicates, n>18.

[0061] Figure 4 [A] Seed germination phenotype of diacetyl-restored ABA-inhibited germination. [B] Quantitative seed germination efficiency, error bar from 4 biological replicates, n>240. Detailed Implementation

[0062] Through extensive and in-depth research, the inventors unexpectedly discovered that diacetyl can effectively (a) delay plant senescence and / or (b) promote ABA-inhibited seed germination. Furthermore, the inventors also unexpectedly discovered that diacetyl can be used to (a) delay plant hormone-induced senescence; (b) restore the phenotype of ABA-inhibited seed germination; and (c) preserve plants. Based on these findings, the inventors completed this invention.

[0063] Diacetyl

[0064] Diacetyl is a small, volatile molecule released by bacteria. Studies have shown that diacetyl can regulate plant growth and immunity; however, its full role in plants remains unclear. Diacetyl is relatively inexpensive, averaging about 90 RMB per milliliter of concentrate. It is widely used as a food additive to enhance flavor in various foods, such as yogurt and bread.

[0065] In this invention, diacetyl is derived from rhizosphere growth-promoting bacteria, such as Bacillus amyloliquefaciens GB03; lactic acid bacteria, such as Lactobacillus harbinensis; cocoa beans, etc.

[0066] Agricultural compositions or formulations

[0067] The active substances of the present invention (such as diacetyl) can be prepared into agricultural formulations, such as solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols, natural and synthetic materials impregnated with the active substances, microcapsules in polymers, and seed coating agents, using conventional methods.

[0068] These formulations can be produced using known methods, such as mixing the active substance with a expander, which can be a liquid, liquefied gas, or solid diluent or carrier, and can be any type of surfactant, i.e., emulsifier and / or dispersant and / or foaming agent. For example, when water is used as the expander, organic solvents can also be used as adjuvants.

[0069] Liquid solvents are generally suitable as diluents or carriers, such as: aromatic hydrocarbons, such as xylene, toluene, or alkylnaphthalene; chlorinated aromatic or chlorinated aliphatic hydrocarbons, such as chlorobenzene, vinyl chloride, or dichloromethane; aliphatic hydrocarbons, such as cyclohexane or paraffins, such as mineral oil fractions; alcohols, such as ethanol or ethylene glycol and their ethers and esters; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, or cyclohexanone; or less commonly used polar solvents, such as dimethylformamide and dimethyl sulfoxide, and water.

[0070] In the case of liquefied gas diluents or carriers, it refers to liquids that become gaseous at normal temperature and pressure, such as aerosol propellants, such as halogenated hydrocarbons, as well as butane, propane, nitrogen, and carbon dioxide.

[0071] Solid carriers can be ground natural minerals such as kaolin, clay, talc, quartz, activated clay, montmorillonite, or diatomaceous earth, and ground synthetic minerals such as highly dispersed silica, alumina, and silicates. Solid carriers for granulation are crushed and graded natural zircon, such as calcite, marble, pumice, sepiolite, and dolomite, as well as granules synthesized from inorganic and organic coarse powders, and granules of organic materials such as sawdust, coconut husks, corncobs, and tobacco stalks.

[0072] Nonionic and anionic emulsifiers can be used as emulsifiers and / or foam forming agents. Examples include polyoxyethylene-fatty acid esters, polyoxyethylene-fatty alcohol ethers, such as alkylaryl polyethylene glycol ethers, alkyl sulfonates, alkyl sulfates, aryl sulfonates, and albumin hydrolysates. Dispersants include, for example, lignin sulfite waste and methylcellulose.

[0073] In the formulation, binders such as carboxymethyl cellulose and natural and synthetic polymers in the form of powders, granules or emulsions, such as gum arabic, polyvinyl alcohol and polyvinyl acetate, can be used.

[0074] Coloring agents such as inorganic dyes, such as iron oxide, cobalt oxide and Prussian blue; organic dyes, such as azo dyes or metallic titanium cyanide dyes; and trace nutrients, such as salts of iron, manganese, boron, copper, cobalt, aluminum and zinc, can be used.

[0075] In this invention, the “agricultural formulation” is typically an agricultural plant growth regulator containing diacetyl as an active ingredient for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination; and an agriculturally acceptable carrier.

[0076] As used herein, "agriculturally acceptable carrier" is a pesticide-acceptable solvent, suspending agent, or excipient for delivering diacetyl of the present invention to plants. The carrier may be liquid or solid. Agriculturally acceptable carriers suitable for use in the present invention are selected from the group consisting of water, buffer solutions, DMSO, surfactants such as Tween-20, or combinations thereof. Any agriculturally acceptable carrier known to those skilled in the art may be used in the present invention.

[0077] The agricultural formulations of the present invention may be contained in commercial formulations or in use formulations prepared from these formulations as a mixture with other substances that (a) delay plant senescence and / or (b) promote ABA-inhibited seed germination. These other substances that (a) delay plant senescence and / or (b) promote ABA-inhibited seed germination include (but are not limited to): small diacetyl molecules, diacetyl salts, BA (6-benzyladenine), vitamin E, gibberellins, and auxins.

[0078] Furthermore, the agricultural formulations of the present invention can also be mixed with synergists and exist in their commercial formulations or in application formulations prepared from these formulations. These synergists are compounds that enhance the activity of the active substance. Since the active substance itself is active, it is not necessary to add synergists.

[0079] The formulation of the agricultural preparation described in this invention can be of various types, as long as the active ingredient can be effectively delivered to the plant. From the perspective of ease of preparation and application, the preferred agricultural preparation is a spray or solution.

[0080] The agricultural formulations described in this invention typically contain 0.0001-99 wt%, preferably 0.1-90 wt%, of the active ingredient of this invention, accounting for 0.0001-99 wt% of the total weight of the agricultural formulation. The concentration of the active ingredient of this invention in commercial formulations or application formulations can vary within a wide range. The concentration of the compound of this invention in commercial formulations or application formulations can range from 0.0000001-100% (g / v), preferably between 0.0001 and 1% (g / v).

[0081] (a) Methods to delay plant senescence; and / or (b) methods to promote ABA-inhibited seed germination.

[0082] The present invention provides a method for delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination, comprising the steps of: applying diacetyl to the plant, or applying a corresponding agricultural composition or formulation.

[0083] Application can be carried out by various known methods, such as by spraying, misting, dusting or sowing diacetyl or agricultural compositions or formulations containing diacetyl on plant leaves, propagation material, or otherwise exposing plants to diacetyl or agricultural formulations containing diacetyl.

[0084] In a preferred embodiment, diacetyl or agricultural compositions or formulations containing diacetyl can also be delivered to the plants by spraying (e.g., aerial spraying) or irrigation.

[0085] The main advantages of this invention include:

[0086] (1) This invention is the first to discover that diacetyl can effectively (a) delay plant senescence; and / or (b) promote ABA-inhibited seed germination.

[0087] (2) The inventors have discovered for the first time that diacetyl can also be used to (a) delay plant senescence induced by plant hormones; (b) restore the phenotype of seed germination inhibited by ABA; and (c) delay the senescence of plants that need to be stored and transported for a long time, so as to achieve the purpose of preservation.

[0088] (3) The method of the present invention is highly safe and extremely low in cost.

[0089] (4) This invention has a wide range of applications and is easy to operate.

[0090] (5) The product of this invention is readily available.

[0091] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions as described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise specified, all materials and reagents used in the embodiments are commercially available products.

[0092] Unless otherwise specified, the experimental methods described in this example are conventional techniques and methods in the art. The experimental reagent kits and consumables used are also conventional reagents and consumables in the art, and can be purchased commercially.

[0093] The Arabidopsis thaliana species involved in the following examples are Col-0, obtained from ABRC (Arabidopsis Biological Resource Center). ABA, methyljasmonic acid and salicylic acid were purchased from Sigma-Aldrich, and diacetyl was purchased from Sangon Biotech. The purity is >97%.

[0094] General Method

[0095] The general method of use for diacetyl is as follows:

[0096] 1. The steps for diacetyl to help leafy green vegetables resist aging are as follows:

[0097] Diacetyl aqueous solution was uniformly sprayed onto the surface of isolated leafy green vegetables (approximately 0.066 micromoles of diacetyl (500 μL 132 μM) are required for one leafy green vegetable plant). The control group was uniformly sprayed with an equal volume of water. The plants of both treatments were placed in a corner at room temperature to observe their anti-aging phenotype.

[0098] 2. The steps for using diacetyl to help seeds resist abscisic acid inhibition of seed germination are as follows:

[0099] Diacetyl agar solution (1% agar content) was used for evaporation treatment of seeds, especially for seeds that have been stored for a long time or are dormant. Taking Arabidopsis thaliana as an example, in a 4-compartment culture plate, seeds and diacetyl were placed in different compartments, with an average of 20 seeds treated with 0.02 μmol diacetyl aqueous solution (100 μL 200 μM). After treatment, the culture dish was wrapped with an airtight membrane and placed under light, and the germination of the seeds was continuously observed and recorded in real time.

[0100] Example 1: Diacetyl specifically delayed the senescence of detached leaves.

[0101] Prepare kitchen paper towels and arrange them into uniformly sized filter paper sheets. Place them in a 9 cm diameter petri dish. Add 2 mL of a 132 μM / L aqueous solution (containing four small molecules, including diacetyl) to the filter paper. The control group was treated with an equal volume of water. We isolated approximately 4-week-old Arabidopsis thaliana leaves and placed them on filter paper containing diacetyl in the petri dish. The petri dishes were wrapped in aluminum foil and placed under low light conditions (in a corner of the room, light intensity approximately 40 μmol / m²·s) or in darkness for 8 days. Each treatment used at least 9 plants. The senescence phenotype of the isolated leaves showed that only the leaves in the petri dish containing diacetyl exhibited a significantly anti-senescence phenotype compared to the control group, while other major small molecule volatiles from the same bacteria did not show a significantly anti-senescence phenotype compared to the control group. The main anti-aging phenotype was that, compared to the control group, the leaves of plants treated with diacetyl were more vibrant green, while the leaves of the control group and other small molecule-treated plants were more golden and wilted. This result shows that diacetyl can specifically delay the aging phenotype of detached leaves under low-light conditions. Figure 1 .

[0102] Example 2: Diacetyl helps preserve plants

[0103] We conducted a laboratory experiment to test the anti-aging effects of diacetyl on fresh bok choy purchased from the market. We found that bok choy, when stored indoors, cannot be kept for too long; generally, yellowing and wilting occur between the 5th and 8th day. We divided the fresh bok choy into two uniform groups (approximately 20 plants per group) under identical indoor low-light storage conditions. One group was sprayed with 2 micromoles of diacetyl (final concentration of diacetyl: 200 μM, total 10 mL), while the other group was not treated with diacetyl but sprayed with an equal volume of water. We observed the leaf senescence of the two groups of bok choy after a certain period (days 5 and 8). Compared to the control group, the bok choy treated with diacetyl showed a greener and healthier leaf condition and was still edible, while the control group's bok choy had wrinkled and yellowed leaves and began to rot (by day 8), rendering it inedible.

[0104] Example 3: Diacetyl delays the senescence of the entire plant.

[0105] Based on the significant delay in senescence of detached leaves by diacetyl, we further examined the anti-senescence phenotype of diacetyl at the whole-plant level. We treated 4-week-old Arabidopsis thaliana seedlings with approximately 42 ml of 132 μM (micromoles per liter) diacetyl per pot. The control group was treated with an equal volume of water. Both diacetyl-treated and untreated plants were placed under low-light conditions (indoor corner, light intensity approximately 40 μmol / m²·s) for 5 days without additional watering. The results showed that after 5 days of low-light treatment, the diacetyl-treated group exhibited a more pronounced anti-senescence phenotype compared to the control group. The main phenotypes were: compared to the control group, the diacetyl-treated plants had greener, more upright leaves, while the control group plants were more golden and their leaves appeared more wilted. This study demonstrates that diacetyl can help achieve anti-senescence at the whole-plant level. Figure 2 A and Figure 2 B.

[0106] Furthermore, experiments of this invention show that diacetyl in the range of 0.01 micromoles (50 μL of 200 μM diacetyl aqueous solution) to 20 micromoles (100 mL of 200 μM diacetyl aqueous solution) can delay the senescence of the whole plant.

[0107] Example 4: Diacetyl delays plant senescence induced by plant hormones

[0108] Our previous study found that diacetyl can effectively achieve anti-senescence in in vitro or whole plants. Next, we explored the effect of ABA on the diacetyl-induced anti-senescence process in plants. By treating plant leaves of approximately 4 weeks old in vitro with ABA and diacetyl (final concentration 132 μM), we found that diacetyl can effectively inhibit ABA-induced plant senescence. We further analyzed the chlorophyll content and ion permeability of the plants, finding that diacetyl can maintain chlorophyll content... Figure 3 B, see reduction in ion permeation efficiency. Figure 3 C. These results indicate that diacetyl can delay ABA-induced plant senescence, see... Figure 3 A.

[0109] Jasmonic acid (JA) has also been reported as an important plant hormone in the senescence process. We also investigated the effect of diacetyl on jasmonic acid-induced plant senescence. The results showed that detached leaves treated with 2 ml of 200 μM jasmonic acid exhibited a more pronounced dark yellow senescence phenotype compared to the control group. However, when leaves treated with jasmonic acid were further treated with 2 ml of 132 μM diacetyl, the leaves showed a more vibrant green phenotype compared to both the control group and the jasmonic acid treatment, demonstrating a significant anti-senescence phenotype. This result indicates that diacetyl can delay the jasmonic acid-induced plant senescence phenotype.

[0110] Salicylic acid (SA) has also been reported as an important plant hormone in the plant senescence process. We also explored the effect of diacetyl on salicylic acid-induced plant senescence. The results showed that detached leaves treated with 2 ml of 2 mM salicylic acid exhibited a more pronounced dark yellow senescence phenotype compared to the control group. However, further treatment of salicylic acid-treated leaves with 2 ml of 132 μM diacetyl resulted in leaves that were a brighter green compared to both the control group and the salicylic acid-treated group, demonstrating a significant anti-senescence phenotype. This result indicates that diacetyl can delay the salicylic acid-induced plant senescence phenotype.

[0111] Example 5: Diacetyl restores ABA-inhibited seed germination efficiency

[0112] Our study found that diacetyl can delay ABA-induced plant senescence. Next, we further explored the role of diacetyl in ABA-inhibited seed germination. We treated Arabidopsis seeds with 2 μM ABA and 200 μM diacetyl, continuously observing the germination efficiency. The results showed that diacetyl did not affect normal seed germination, but it significantly increased the seed germination rate under ABA conditions. This result indicates that diacetyl can restore ABA-inhibited seed germination. Figure 4 A and Figure 4 B.

[0113] We also observed the recovery of seed germination in crops such as corn and rice by treating them with ABA and then adding diacetyl exogenously. We found that compared to the control group, 5 μM / L ABA effectively inhibited the germination of corn and rice seeds. However, the experimental group with 200 μM diacetyl exogenously added showed a significant recovery in seed germination efficiency. The seed germination results of these crops indicate that diacetyl can also effectively restore the ABA-inhibited seed germination phenotype.

[0114] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A use of diacetyl, characterized in that, For (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination; or for the preparation of a composition or formulation for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination.

2. The use as described in claim 1, characterized in that, The formulation or composition is also used for one or more purposes selected from the group consisting of: (a) Delaying plant hormone-induced senescence; (b) Restoring the phenotype of ABA-inhibited seed germination; (c) Preserving plants.

3. The use as described in claim 1, characterized in that, The plants include those that have been stored in the dark and / or in low light conditions.

4. The use as described in claim 1, characterized in that, The plants mentioned include detached plants.

5. The use as described in claim 1, characterized in that, The plants mentioned include agricultural plants, horticultural plants, or forestry plants.

6. A composition, characterized in that, include: (i) Diacetyl; (ii) Other substances used to (a) delay plant senescence; and / or (b) promote ABA-inhibited seed germination; (iii) An agriculturally acceptable carrier; The other substances used to (a) delay plant senescence and / or (b) promote ABA-inhibited seed germination are BA, vitamin E, gibberellin, or auxin.

7. The composition according to claim 6, characterized in that, The dosage form of the composition is selected from the group consisting of: solutions, emulsions, suspensions, powders, foams, pastes, granules, and aerosols.

8. Use of the composition according to claim 6, characterized in that, Used to (a) delay plant senescence; and / or (b) promote ABA-inhibited seed germination.

9. A method for (a) delaying plant senescence; and / or (b) promoting ABA-inhibited seed germination, characterized in that, Including the following steps: The plant may be treated with diacetyl or the composition of claim 6.

10. The method as described in claim 9, characterized in that, The method is performed under conditions of light avoidance and / or low light.

11. The method as described in claim 10, characterized in that, The term "weak light" refers to light intensity ≤ 60 micromoles / square meter / second.

12. The method as described in claim 9, characterized in that, The dosage applied is 0.01-400 micromoles per plant.

13. The method as described in claim 9, characterized in that, The applied dose is 5-400 micromoles per plant.

14. A method for preserving plants, characterized in that, The method includes applying diacetyl or the composition of claim 6 to the plant.

15. The method as described in claim 14, characterized in that, The method is performed under conditions of light avoidance and / or low light.

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