Fertilizer composition comprising zeolite and basic l-amino acid

Abstract

The present invention relates to promoting the growth of at least one plant by providing a fertilizer composition comprising at least one zeolite having adsorbed in its pores at least one basic L-amino acid; adding the fertilizer composition to a plant growth medium; and plants are cultivated in the soil. The present invention will provide nitrogen released from the fertilizer composition during plant cultivation at a rate that facilitates plant utilization. The invention also relates to a fertilizer composition, optionally in combination with a plant growth medium to constitute a growth promoter material. The invention is advantageously used to reduce the resources required to fertilize slowly growing plants, wherein repeated supply of nutrients and leakage of nitrogen cause huge cost to the society.

Description

[0001] This application is a divisional application of patent application No. 201780039053.6, filed on June 22, 2017, entitled "Fertilizer Composition Containing Zeolite and Alkaline L-Amino Acids". Technical Field

[0002] This invention relates to fertilizer compositions and their use in providing nitrogen release to plants. More specifically, this invention relates to a method for providing nitrogen to growing plants at a rate corresponding to the nitrogen requirements of the plants. Background Technology

[0003] Methods for improving soil and / or growing conditions have been used in principle since the very first days of agriculture and horticulture. Starting with a very limited understanding of the mechanisms, it was recognized that waste from livestock such as dairy cows could improve the growth of field crops. Because nitrogen, potassium, and phosphorus were identified as key components needed to effectively fertilize soil, commercial formulations became widely available, and the principle of "more is less" was generally applied for decades, leading to the now well-known effects of over-fertilization. While formulations including nitrogen, potassium, and phosphorus, along with various other mineral nutrients, still constitute the standard in most plant cultivation, meticulous research is constantly being conducted on fertilizer compositions that provide plants with the substances they need for optimal growth. Specially designed compositions for certain plants have been developed, and different forms, such as liquid and dry formulations, are also available to balance desired growth, applicability, and minimal environmental impact.

[0004] One approach to reducing the harmful environmental impacts of fertilizers, particularly the loss of mineral nutrients to recipient ecosystems, is to develop compositions that provide slow or delayed release of the active ingredients. Such compositions are commonly referred to as sustained-release or controlled-release formulations.

[0005] Coating mineral nutrient salts has been proposed as a method to slow down this release. However, as a common mechanism, coatings typically delay all release rather than slow the release rate of the nutrients they encapsulate. Thus, in the early stages, the coating may prevent the release of any and all nutrients, and once the formulation is "opened" or the coating has been consumed, the nutrients are immediately and fully available. Therefore, at that point, the released nutrients will either be utilized by the cultivated plants or, if in greater quantities than needed, will leak into the environment. Therefore, a common challenge with coating technology is to provide a time-prolonged release at a rate suitable for the needs of the cultivated plants.

[0006] WO 2015 / 066691 (University of Florida Research Foundation) relates to a slow-release fertilizer composition in which a graphene oxide film is used to delay release. More specifically, the fertilizer composition comprises a plurality of fertilizer particles and a layer of reduced graphene oxide disposed on the surface of each particle. The fertilizer particles may contain one or more of nitrogen, phosphorus, potassium, calcium, magnesium and sulfur, boron, chlorine, copper, iron, manganese, molybdenum, zinc and nickel, at least one of which is in the form of a salt and is available for reducing graphene oxide. The coating technology described herein is claimed to offer great promise for environmentally friendly controlled-release fertilizers for crop production.

[0007] An alternative method for optimizing nutrient release to plants is proposed: the production of its complex. WO 2016 / 035090 (Chaudhry) relates to such fertilizer compositions and methods of their preparation. More specifically, a multifunctional organic-biocomplex composition is described, comprising nutrient sources such as nitrogen, phosphorus, and potassium, and phosphopeptides, such as phosphopeptides comprising an organic acid: a biocomplex agent and a phosphorus source. The biocomplex agent can be a peptide, amino acid, or hydrolyzed protein. The described nitrogen-cation complex is proposed to improve efficiency compared to conventional fertilizers using nitrogen from urea (which is claimed to evaporate rapidly). In the agricultural field, microporous aluminosilicate materials (called zeolites) have been proposed to have soil-strengthening properties due to their high mineral content. For example, Frederick A. Mumpton (in La roca magica: Uses of natural zeolites in agriculture and industry; Proc. Natl. Acad. Sci. USA, Vol. 96, pp. 3463–3470, March 1999, Colloquium Paper) proposed adding natural zeolite clinoptilite to the soil along with standard fertilizers to delay its ammonium release.

[0008] Because zeolites are cation exchangers, they can be advantageously used in water purification, particularly in water softening. In sodium zeolite softening, water containing scaling ions (such as calcium and magnesium) flows through a resin bed, where hard ions are exchanged for sodium ions, which diffuse into a large volume of the aqueous solution. Hardness-free water can then be used in boiler feedwater, reverse osmosis systems, and various chemical processes.

[0009] Zeolites have also been studied as alternatives to organic ion exchangers for the separation of amino acids, particularly in the preparation of amino acids through extraction, synthesis, or fermentation.

[0010] FC Nachod (in Ion Exchange: Theory and Application, Elsevier, December 2, 2012, Chapter II (Separation of basic amino acids) shows that neutral and acidic amino acids can be effectively extracted from zeolite using standard extraction procedures and media, but basic amino acids, especially arginine and lysine, bind so strongly to zeolite that they are more or less immobile.

[0011] Furthermore, Nelson et al. pointed out that only strong acid (2N HCl) could effectively extract arginine and lysine bound to the Delcaso (a type of zeolite) column, while all other amino acids could be effectively extracted using pyridine.

[0012] Finally, WO 2005 / 075602 (Balance Agri-Nutrients Ltd) relates to fertilizer compositions, and more specifically to compositions in the form of granular zeolite containing at least one nitrification inhibitor. Therefore, the purpose of WO 2005 / 075602 is to reduce the loss of nitrates to the environment by inhibiting the conversion of ammonium to nitrites and nitrates. An exemplary fertilizer composition according to WO 2005 / 075602 comprises 10% fertilizer (e.g., urea), 10-70% zeolite, and 1-45% nitrification inhibitor.

[0013] However, given the many different plants and growing conditions in consumer use and at the commercial scale, and the growing need to reduce the environmental impact of fertilizer compositions, alternative methods and products are still needed to support the effective cultivation of plants. Summary of the Invention

[0014] One object of the present invention is to provide a method for fertilizing plant growth media in a manner that minimizes nitrogen loss.

[0015] Another object of the present invention is to provide a method that has the effect of fertilizing plants over an extended period of time.

[0016] A specific object of the present invention is to provide a form of applying organic nitrogen to growing plants that provides protection against nitrogen, for example, by preventing microbial utilization.

[0017] Another object of the present invention is to provide on-demand fertilization of plants, wherein plant activity controls the release of nitrogen from the fertilizer composition.

[0018] The above-mentioned objectives can be achieved as described in the appended independent claims. Other embodiments, details, and advantages of the invention will become apparent from the dependent claims and the following detailed description and experimental section.

[0019] The term “plant” is used in a broad sense in this article to refer to a single plant or a class of plants.

[0020] The term “promote” plant growth is used broadly in this article, including providing or enhancing, that is, improving the growth of any or all parts of a plant.

[0021] The term "amino acid" as used in this article includes amino acid derivatives or modified forms thereof.

[0022] The term "zeolite" refers to microporous aluminosilicate minerals, including both natural and synthetic forms of this material.

[0023] In the context of zeolites used according to the present invention, the term "adsorption" is used in its broadest sense to include any chemical interaction and / or binding principle that provides a connection.

[0024] The term "mushroom" is used in this article to refer to the fleshy, spore-bearing fruiting body of a fungus, which typically grows on the surface of soil or other media used for plant growth.

[0025] The term "field root" is defined as roots that grow in the field, rather than in the peat filler of a pot.

[0026] The term “plant growth medium” is used in its broadest context herein and may include, for example, peat, clay, various compositions of sand, soil and any combination thereof that are considered suitable or desirable for plant cultivation. Attached Figure Description

[0027] Figure 1 The growth of barley loaded with arginine-containing zeolite of the present invention is shown with (right) and without (left).

[0028] Figure 2 The growth of lettuce loaded with arginine-containing zeolite of the present invention is shown with (right) and without (left).

[0029] Figure 3 These are photographs illustrating the growth of lettuce and barley using the unfertilized control (left) and the arginine-zeolite of the present invention (right).

[0030] Figure 4A -B shows the biomass (4A) and total nitrogen content (4B) of the branch growth of zeolite-loaded arginine seedlings of the present invention with (right) and without (left) arginine.

[0031] Figure 5 These are photographs illustrating the growth of pine seedlings in a control group without fertilizer (left) and in a pine seedling using the arginine-zeolite of this invention (right).

[0032] Figure 6The germination of pine seeds on zeolite loaded with arginine (left) or pine seeds using commercially available fertilizer (right) is shown.

[0033] Figure 7 The growth of pine seedlings on zeolite loaded with arginine (right) and pine seedlings on control (left) without zeolite are shown.

[0034] Figure 8 These are photographs illustrating the growth of pine seedlings with mycorrhizae (left) or without mycorrhizae (right) using zeolite loaded with arginine according to the present invention.

[0035] Figure 9A -B shows the growth of biomass and total nitrogen in the needles of pine seedlings with mycorrhizae (right) or without mycorrhizae (left) on zeolite loaded with arginine.

[0036] Figure 9C This is a set of four photos of pine seedlings with mycorrhizae (1 and 2) and without mycorrhizae (3 and 4).

[0037] Figure 10 This describes the mass loss during the extraction process after 18 days of growth promotion according to the present invention, compared to growth using existing fertilizers.

[0038] Figure 11A-C This describes the total biomass of field roots in Norway spruce and twisted pine seedlings as described in Example 7.

[0039] Figure 12A -B shows increased branch growth in Norway spruce and twisted pine with (right) and without (left) arginine-loaded zeolite obtained according to Example 7 below.

[0040] Figure 13 The dry weight of grass fragments from fairway turf treated with the arginine-loaded zeolite of the present invention is shown compared to different prior art fertilizers.

[0041] Figure 14 The nitrogen (N) recovery rate in fairway turf fragments is shown after a 6-week response time to different fertilizers. Detailed Implementation

[0042] This invention relates to methods and products for fertilizing and cultivating plants in a manner that minimizes nitrogen leakage into the environment. More specifically, the invention enables fertilized plants to obtain and utilize the amount of nitrogen required for their nitrogen metabolism. Therefore, the invention can be considered as relating to on-demand fertilization, wherein the activity of the fertilized plant controls the release of nitrogen from the fertilizer composition.

[0043] A first aspect of the present invention is a method for promoting the growth of at least one plant, the method comprising: a) Provide a fertilizer composition comprising at least one zeolite in which at least one basic L-amino acid is adsorbed in its pores; b) Add the fertilizer composition to the plant growth medium related to planting; c) Provides nitrogen release from the fertilizer composition during subsequent cultivation of the plant.

[0044] Fertilizer compositions may optionally contain other growth-promoting components, such as those known in the art.

[0045] In step b), adding the fertilizer composition “related to planting” includes adding the fertilizer composition within a limited time before, simultaneously with, and / or within a limited time after planting. In this context, “planting” can include adding seeds, seedlings, or plants to a plant growth medium.

[0046] In step c), the technician will be able to easily determine commonly used methods and measures to provide nitrogen release. For example, adjusting the pH of the growth medium or simply providing humidity by watering can constitute the measures in step c). In some cases, if the plant is already being cultivated under suitable growing conditions, step c) can be simply constructed to maintain the plant under such suitable conditions.

[0047] According to the present invention, fertilized plants will obtain organic nitrogen, i.e., nitrogen derived from amino acids, which has been shown in the prior art to have different and in many cases beneficial effects on plant growth compared to inorganic nitrogen derived from, for example, ammonium fertilizers.

[0048] The basic L-amino acid may be selected from L-arginine; L-lysine; and L-histidine. In one embodiment, the basic L-amino acid is L-arginine and / or L-lysine. In this context, it should be understood that the amino acids used in this invention may include modified forms of basic L-amino acids, as long as they have the property of providing nitrogen to plants as described herein. Basic L-amino acids are available from commercial sources. Fertilizer compositions may include mixtures of basic L-amino acids.

[0049] The zeolites of this invention may comprise any natural and / or synthetic microporous aluminosilicate minerals having a three-dimensional structure comprising coplanar AlO4 and SiO4 tetrahedra. Those skilled in the art will understand that the higher the aluminum content of the zeolite, the more negative charges it can be used for ion exchange processes, and therefore the higher the content of basic L-amino acids per zeolite.

[0050] In one embodiment, the zeolite is a natural zeolite. In a specific embodiment, the zeolite is selected from analcime; chalcogenide; clinoptilolite; bufosite; octahedralite; magnesium alkali zeolite; flaky zeolite; turbidite; mordenite; calcium cruciformite; linde A; and linde B. In an advantageous embodiment, the zeolite is clinoptilolite, which is a zeolite mainly comprising SiO2 and Al2O3 and small amounts of CaO and K2O, or a mixture of different zeolites including clinoptilolite.

[0051] Modified zeolites can be used, provided they have the cation exchange capacity described herein.

[0052] In this context, the term "zeolite" should be understood to refer to multiple zeolite entities of the same kind or form.

[0053] The fertilizer compositions of the present invention can be prepared according to previously proposed methods, see, for example, Krohn et al. discussed above. Those skilled in the art will understand that the adsorption of amino acids to zeolites will involve ion exchange, but may also include other mechanisms, such as hydrogen bonding. Zeolites or zeolite mixtures may be provided in granular, particulate, or any other suitable form.

[0054] In step a, it is advantageous to wash the zeolite after adsorbing the amino acids to avoid any potential toxicity caused by the immediate release of a relatively large amount of nitrogen if nitrogen loosely adhered to the outside of the zeolite is left. Therefore, washing prevents premature nitrogen release, i.e., release unrelated to plant activity.

[0055] While it is well known that plants can exude chemicals from their roots to improve the acquisition of mineral nutrients, the difficulty of releasing basic amino acids from zeolite has also been extensively documented, see FC Nachod and Nelson et al., as discussed in the background section above. Therefore, it can be expected that nitrogen from zeolite adsorbed with basic amino acids such as arginine and lysine will be difficult or even impossible to enter the plant, and thus cannot be used as a fertilizer in plant cultivation. Therefore, the discovery of this invention is quite unexpected: the plant itself is actually able not only to release nitrogen as basic amino acids adsorbed in zeolite, but also to control the rate of nitrogen release according to their activity.

[0056] Those skilled in the art will understand that, according to the present invention, the properties and amounts of amino acids, as well as the appropriate selection of zeolites, can be used as a means of optimizing fertilizer compositions for specific plants and / or growth conditions.

[0057] Therefore, the amount of amino acids, i.e., the amino acid loading on the zeolite, should be adjusted according to the plant species, the plant growth medium, the humidity in which the plant will grow, and the expected or desired growth rate or growth period. In this application, adsorption to zeolite sometimes refers to the loading or charge on the zeolite.

[0058] As discussed above, the on-demand functionality of the present invention can be used for the cultivation of any plant, and as shown in the experimental section, zeolite can have any nitrogen content because the plant itself will direct the release of nitrogen. However, as will be discussed below, specific growth materials can be designed to be optimal in various aspects of a particular environment.

[0059] Therefore, in one embodiment, the nitrogen loading from the basic L-amino acids in the zeolite is at least 1%, for example at least 2% or at least 3%, based on the total weight of the zeolite loaded per load. In one embodiment, the nitrogen loading from the basic L-amino acids in the zeolite is up to 10%, based on the total weight of the zeolite loaded per load. Useful ranges may be 1-3%; 2-3%; 1-10%; 2-10%; or 3-10% nitrogen from the basic L-amino acids, based on the total weight of the zeolite loaded per load. In this context, the term "load" refers to the amount adsorbed into the zeolite pores by ion exchange and other optional binding mechanisms. Those skilled in the art will be able to adjust the appropriate nitrogen loading per weight or volume of zeolite according to various factors, such as the container or environment in which the plants will grow. For smaller containers, a higher loading may be advantageous, while in other cases, larger soil volumes may require a lower nitrogen loading per zeolite or work well with a lower nitrogen loading per zeolite.

[0060] This invention can be used, for example, in the cultivation of slow-growing plants that require fertilizer during their extended growth period. By using this invention, such plants can be successfully cultivated with less fertilizer additives and less nitrogen leakage into the environment compared to existing technologies. Therefore, this invention enables the addition of large amounts of fertilizer for the long-term and continuous nutrition of slow-growing plants.

[0061] In one implementation, the plants to be fertilized are conifers, such as members of the order Pinales, including members of the family Cupressaceae, such as *Cupressus* spp., *Juniperus* spp., *Sequoia* spp., and *Sequoiadendron* spp.; members of the family Taxaceae (*Taxus* spp.); and members of the family Pinaceae, such as *Abies* spp., *Cedrus* spp., *Larix* spp., *Picea* spp., *Pinus* spp., *Pseudotsuga* spp., and *Tsuga* spp. In one favorable implementation, the plant to be fertilized is a member of the genus *Pinus* or *Picea*, such as Scots pine (*Pinus sylvestris*), twisted pine (*Pinus contorta*), or Norway spruce (*Picea abies*).

[0062] In another implementation, the plants to be fertilized are broad-leaved trees, including hybrids and cultivars, such as locust (Acacia spp.), alder (Alnus spp.), birch (Betula spp.), hornbeam (Carpinus spp.), hickory (Carya spp.), chestnut (Castanea spp.), beech (Fagus spp.), walnut (Juglans spp.), oak (Quercus spp.), ash (Fraxinus spp.), poplar (Populus spp.), aspen (Populus), and willow (Salix spp.). Eucalyptus (Eucalyptus spp.), Platanus (Platanus spp.), Acer (Acer spp.), Mahogany (Swietenia spp.), and Liquidambar (Liquidambar spp.).

[0063] In the specific implementation plan, the plants to be fertilized are woody plants whose leaves can be eaten as leafy vegetables, including species of the genera Adansonia, Aralia, Moringa, Morus, and Toona.

[0064] In another implementation, the plants to be fertilized are fruit-bearing plants, including hybrids and cultivars, such as apples (Malus spp.), plums (Prunus spp.), pears (Pyrus spp.), citrus (Citrus spp.), lemons (Citrus spp.), kiwifruit (Actinidia spp.), cherries (Prunus spp.), grapevines (Vitis spp.), figs (Ficus spp.), and bananas (Musa spp.). Other fruit-bearing plants include shrubs such as raspberries or blueberries (Vaccimium spp.) and bromeliads such as pineapples.

[0065] Orchids such as vanilla or phalaenopsis, succulents such as cacti (Cactaceae), and Euphorbia (Euphorbiaceae) are other examples of slow-growing plants that can be fertilized according to the present invention.

[0066] The method of the present invention can also be used to cultivate fast-growing plants that typically require nitrogen during a short growth period. Therefore, in one embodiment, the plant is an annual or biennial, and the loading of nitrogen derived from basic L-amino acids in the zeolite is about 1-10%, calculated based on the total weight of the loaded zeolite.

[0067] In one implementation, the plants to be fertilized are monocotyledonous plants, including hybrids and cultivars, selected from barley (Hordeum vulgare), maize (Zea mays), rice (Oryzasativa), sorghum (Sorghum spp.), wheat (Triticum), millet (Eleusine coracana), foxtail millet (Setaria italica), black millet (Pennisetum glaucum), millet (Panicum miliaceum), oats (Avenasativa), black wheat, wheat hybrids, crabgrass (Digitaria), onion (Allium spp.), and pineapple (Ananas). spp.), rye (Rye (Secale cereale)), amaryllis, bamboo (Bambuseae), banana (Musaceae), round-leaved bellflower (Hyacinthoides), reed, daffodil (Narcissus), ginger family (Zingiberaceae), butterfly flower (Iris), lily (Lilium), orchid (Orchidaceae), palm (Arecaceae), sugarcane (Saccharum spp.) and tulip (Tulipa).

[0068] In one advantageous implementation, the plant to be fertilized is a grass, such as a member of the Poaceae family, including hybrids and cultivars selected from: bluegrass (Poa spp.), creeping bentgrass (Agrostis spp.), ryegrass (Lolium spp.), fescue (Festuca spp.), Calamogrostis spp., hairgrass (Deschampsia spp.), tufted fescue (Festuca paradoxa spp.), Zoysia (Zoysia spp.), bermudagrass (Cynodon spp.), Stenotaphrum secundatum (Stenotaphrum secundatum), Paspalum (Paspalum spp.) The beneficial grasses to be fertilized according to the present invention are those from the genera *Poa* or *Festuca*.

[0069] In another embodiment, the plant to be fertilized is a dicotyledonous plant, which includes hybrids and cultivars selected from the following: alfalfa (Medicago sativa), medicago truncatula, legumes (Phaseolus), beets (Beta vulgaris), buckwheat (Fagopyrum esculentum), carob (Onia siliqua), chickpea (Cicer arietinum), cotton (Gossypium spp.), cucumber (Cucumis sativus), pea (Pisumsativum), peanut (Arachis hypagaea), pepper (Piper spp.), and potato (Solanum tuberosum). The following are considered suitable plants for fertilization according to the present invention: *Glycine max* (soybean), *Spinaciaoleracea* (spinach), *Lactuca spp.* (lettuce), *Cucurbita* (cucurbita), sunflower (sunflower), tomato (tomato), and wild soybean (wild soybean). Additionally, herbs such as basil (Ocimum spp.) and oregano (Origanum spp.), or ornamental plants belonging to the clade Rosids (e.g., *Geranium spp.*), can also be used.

[0070] As can be seen from the above, a fertilizer composition containing zeolite in which basic L-amino acids are adsorbed can be added to the growth medium before, after, or simultaneously with placing the seeds, plants, or seedlings in the growth medium. One advantage of this invention is the ability to prepare pre-fertilizer materials that provide fertilizer that will sustainably extend the growth period and will be supplied to the cultivated plants at a rate corresponding to the plant's own needs, i.e., corresponding to the nitrogen requirements of the plant associated with its growth activities. In this case, those skilled in the art will understand that "corresponding" is an approximation, and some excess nitrogen will still be released into the environment. However, from a leakage point of view, any such release will be small enough to be negligible.

[0071] A second aspect of the invention is a fertilizer composition comprising at least one zeolite in which at least one basic L-amino acid is adsorbed in its pores, optionally together with other growth-promoting components. Other growth promoters may be selected from potassium, phosphorus, metal ions, vitamins, and minerals. Furthermore, the fertilizer composition of the invention may include commonly used additives to provide a suitable physical form, such as granular or particulate material. A suitable particle size may depend on the contents of the plant to be cultivated therein and can be readily determined by those skilled in the art.

[0072] Therefore, in one embodiment, the present invention is a growth support material comprising any conventional medium for plant growth combined with at least one zeolite, wherein at least one basic L-amino acid is adsorbed in the pores of the zeolite. Thus, due to the content of organic nitrogen adsorbed in the pores of the zeolite, this embodiment can be considered a pre-fertilized growth material.

[0073] All the details, embodiments, and examples provided above regarding the invention, such as amino acids and their amounts, zeolites, plants, and growth media, are equally applicable to this second aspect.

[0074] Growth support materials consisting of any conventional growth medium combined with zeolite, in which amino acids are adsorbed in the pores, can be provided in any suitable form. Therefore, it can be provided in the form of microparticles or granules in bags of growth medium, or in jiffy pots. The materials of this invention can be provided in a form suitable for private use or in a larger form more suitable for commercial scale. Some forms including this invention are particularly suitable for automation.

[0075] In one embodiment, the growth support material of the present invention is provided in a biodegradable container. The biodegradable container may be a peat pot or the like.

[0076] In another embodiment, the growth support material of the present invention is a mat arranged for planting seeds or seedlings. In this case, such a mat can be compressed and optionally be a dry growth medium.

[0077] The growth support material of the present invention can be used in any environment where nitrogen is expected to be released at a rate corresponding to the growth needs of the cultivated plant, such as for agricultural or horticultural purposes, for consumer use in homes or gardens, in greenhouses, and in outdoor tree plantations.

[0078] A third aspect of the invention is the use of at least one zeolite as a fertilizer, wherein at least one basic L-amino acid is adsorbed in the pores of the zeolite.

[0079] A third aspect of the invention also includes the use of the growth support material as described above in the fertilization and cultivation of at least one plant.

[0080] All the details, implementation schemes, and examples provided above regarding, for example, amino acids and their amounts, zeolites, plants, forms, and growth media, are equally applicable to this third aspect.

[0081] In an advantageous embodiment of the use of the invention, the plant is a perennial plant and the loading of nitrogen derived from the basic L-amino acid in the zeolite is about 1-3%, calculated based on the total weight of the loaded zeolite.

[0082] In one specific embodiment of the invention, the plant is a coniferous tree, such as a member of the Pinaceae family, such as the genus Pinus or the genus Picea.

[0083] In one embodiment, the method of the present invention, the fertilizer composition of the present invention, or the growth-promoting material of the present invention is used in the cultivation of at least one mycorrhizal plant.

[0084] In one embodiment, the plant is able to form a symbiotic relationship with the fungus. The fungus may be able to form fruiting bodies, such as those used as food and for cooking. Therefore, the present invention can be advantageously used for the large-scale cultivation of fungal fruiting bodies in the food industry and of any mushroom.

[0085] Fungi can also enhance the performance of cultivated plants or seedlings. Therefore, this invention enables plants to grow rapidly while maintaining or improving a symbiotic relationship with one or more mycorrhizal fungi. Once planted in soil, such as in field environments like farmland or reforestation areas, these mycorrhizal fungi will subsequently and over an extended period improve the performance of the plants or seedlings.

[0086] The present invention includes any combination of embodiments described in the context of the specific aspects above, provided that those skilled in the art can recognize, for example, that such combinations can satisfy one or more of the objectives of the present invention. Attached Figure Description

[0087] Figure 1 The biomass of each barley plant, expressed in grams, is shown. The bars on the left represent the control without zeolite; while the bars on the right represent the results of growth of the present invention with zeolite loaded with arginine. More specifically, the nitrogen content was 2% (in the form of L-arginine), totaling 20 mg N per pot. Plants were grown in pots filled with soil and harvested after 8 weeks, as described in more detail in Example 3. The bars represent the mean ± standard deviation (n = 18–21) of the dry weight of the whole plant (including roots and branches). Figure 1 As shown, the biomass obtained when using the arginine-loaded zeolite of the present invention is almost twice that of the control. Figure 2The biomass, expressed in grams, is shown for the dry weight of each lettuce plant. The bars on the left represent the control without zeolite; while the bars on the right represent the results of growth of the present invention with zeolite loaded with arginine. More specifically, the nitrogen content was 2% (in the form of L-arginine), totaling 20 mg N per pot. Plants were grown in pots filled with soil and harvested after 8 weeks, as described in more detail in Example 3. The bars represent the mean ± standard deviation (n = 18–21) of the dry weight of the whole plant (including roots and branches).

[0088] Figure 3 This describes the result obtained according to Example 3. Figure 1 and 2 The photos show increased biomass. More specifically, Figure 3 The growth of lettuce (Lactuca) and barley (Haloxylon) on unfertilized control (left) and zeolite loaded with arginine (20 mg N) (right) is shown. The growth of the plant on the right was significantly enhanced by the addition of the amino acid-loaded zeolite of the present invention.

[0089] Figure 4 shows the branch growth and total nitrogen content in the needles of *Pinus tabuliformis* seedlings with and without arginine-loaded zeolite (2% nitrogen content in the form of L-arginine, totaling 40 mg N per pot) as described in Example 4. The plants were pre-grown in a conifer nursery, and the arginine-loaded zeolite was added to the rootstock before planting in the field. The plants were harvested after one growing season (3 months), and the dry weight of the stems and the total nitrogen content in the needles were determined. Figure 4A In this study, growth is described by branch biomass, where each branch represents the mean ± standard deviation (n = 25) of the dry weight of the entire plant (roots and branches). Clearly, the arginine-loaded zeolite used according to this invention results in a significant increase in branch biomass. Figure 4B The total nitrogen content in the needles shows evidence of pine trees utilizing nitrogen from zeolite.

[0090] Figure 5 These are photographs illustrating the growth of pine seedlings coated with arginine-loaded zeolite in a field trial, comparing the unfertilized control (left) and arginine-zeolite (40 mg N) (right) as described in Example 5. Clearly, the arginine-loaded zeolite used according to the present invention has a growth-enhancing effect.

[0091] Figure 6 The illustration shows how germination rates of pine seedlings (Pinus sylvestris) fertilized with arginine-loaded zeolite of the present invention (left) were almost 100%, while commercially fertilized (right) seedlings had not yet shown any germination. The latter were fertilized with an amino acid-based non-zeolite fertilizer.

[0092] Figure 7The growth of pine seedlings (European Scots pine) with or without zeolite loading (2% nitrogen in the form of L-arginine, totaling 20 mg N per pot) is shown. The plants were grown in pots filled with peat moss and harvested after 12 weeks. Bars represent the mean and standard deviation (n = 10) of the dry weight of the whole plant (roots and branches). The significant differences in biomass of the pine trees cultivated according to the invention clearly illustrate the effects of the invention.

[0093] Figure 8 The images show the growth of pine seedlings (European red pine), and more specifically, photographs of their growth with (left) or without (right) mycorrhizal growth using zeolite loaded with arginine. The seedlings with mycorrhizal growth are significantly larger than those without, further illustrating the effectiveness of the invention.

[0094] Figure 9 shows the growth of pine (Scottish red pine) seedlings with or without mycorrhizal roots on zeolite loaded with arginine, and the total nitrogen content in the needles (2% nitrogen in the form of L-arginine, 20 mg N per pot). Plants were grown in pots filled with peat moss and harvested after 12 weeks. Bars represent the mean and standard deviation (n = 10) of the dry weight of the whole plant (roots and branches). More specifically, Figure 9A The dry biomass of pine trees with and without mycorrhizae (left) and with mycorrhizae (right) is shown; while Figure 9B The images show total nitrogen (mg N / dry weight) in needles without mycorrhizae (left) and with mycorrhizae (right), serving as evidence of pine trees utilizing nitrogen from zeolite. Clearly, the zeolite-bound mycorrhizae loaded with arginine according to the present invention result in a significant increase in biomass.

[0095] Figure 9C This is a set of four photographs showing the growth of pine seedlings with and without mycorrhizal addition. More specifically, boxes 1-4 show the growth of pine seedlings with mycorrhizal addition in area 1; the bottom of the box with mycorrhizal addition in area 2; the growth of pine seedlings without mycorrhizal addition in area 3; and the bottom of the box without mycorrhizal addition in area 4. It should be noted that the pine plantlets in area 1 are very green and vigorous, while the plantlets in area 3 have a pale yellow hue and suffer from nitrogen deficiency.

[0096] Figure 10 The following example 2 illustrates the mass loss during extraction 18 days after growth promotion according to the present invention, compared to growth using existing fertilizers (Osmocote). In summary, Figure 10 As shown, even after washing with water (H2O) (left), 50 mM calcium chloride (CaCl2) (middle), and 50 mM oxalic acid (right), arginine strongly binds to zeolite.

[0097] Figure 11 illustrates the total biomass growth of field roots in Norway spruce and twisted pine seedlings as described in Example 7. More specifically, Figure 11A is a photograph illustrating how field roots develop, see arrows. In Figures 11B and 11C, field root biomass is shown in gram dry weight per plant. Reference seedlings do not receive nitrogen. Field root growth is promoted according to the invention by using zeolite loaded with arginine.

[0098] Figure 12A -B respectively show the increased branch growth of *Pinus tabuliformis* and *Picea spruce* when treated with the zeolite loaded with arginine according to the present invention. More specifically, Figure 12A The dry weight of the branches of the twisted pine is shown, while Figure 12B The dry weight of Norway spruce branches is shown. According to Example 7 below, control seedlings do not receive nitrogen. This figure illustrates that, compared to the control, providing zeolite loaded with arginine to the seedling roots according to the present invention has superior long-term effects on field root growth (Fig. 11) and branch growth (Fig. 12).

[0099] Figure 13 The dry weight of grass fragments collected weekly from 20 mm above the sandy growing substrate during the 6-week response period for different fertilizers is shown in Table 1 below. The dry weight of the pretreated fragments was determined at week 0. N = 4. Error band = standard deviation.

[0100] Figure 14 The weekly nitrogen (N) recovery rates in fairway turf fragments over a 6-week response period with different granular fertilizers are shown in Table 1 below. The dry weight of the pretreated fragments was determined at week 0. N = 4. Error band = standard deviation. Detailed Implementation

[0101] The embodiments of the present invention are for illustrative purposes only and should not be construed as limiting the invention as defined by the appended claims. All references cited herein, both below and elsewhere in this application, are incorporated herein by reference.

[0102] General methods for preparing zeolites loaded with L-amino acids Prepare an aqueous solution of L-amino acids in alkaline or HCl form and adjust the pH to 3-9. Before adsorption, rinse the zeolite in pure water to remove dust particles and other impurities. Add the L-amino acid solution to the rinsed zeolite and stir continuously at room temperature for 2-4 days. Then, thoroughly wash the zeolite in pure water and dry it.

[0103] Example 1: Preparation of zeolites loaded with basic amino acids Example 1(a): Preparation of arginine-loaded zeolite A 0.14 M L-arginine solution was prepared by dissolving 60 g of arginine in 2500 mL of pure water. The pH of the solution was adjusted to 3.5 by adding concentrated HCl. Natural clinoptilolite-type zeolite was obtained from Incal Mineral (Izmir, Turkey) and impurities were removed by rinsing with water. 600 g of zeolite was added to the arginine solution, and the solution was kept at room temperature on a rotary table for 3 days. After removing the L-arginine solution, the zeolite was washed three times with pure water and dried in an oven at 65 °C for 24 hours. The nitrogen concentration in the loaded zeolite was determined using a DeltaV isotope ratio mass spectrometer and a Flash EA 2000 elemental analyzer (both provided by Thermo Fisher Scientific).

[0104] Example 1(b): Preparation of lysine-loaded zeolite A 0.2 M L-lysine solution was prepared by dissolving 7.3 g of L-lysine hydrochloride (98%, Sigma) in pure water. The pH of the resulting solution was adjusted to 8.5 by adding 5 M sodium hydroxide solution.

[0105] Obtain clinoptilolite-type natural zeolite from Incal Mineral (Izmir, Turkey). Add 10 g of zeolite to each of four 50 mL polypropylene test tubes.

[0106] The zeolite was rinsed three times in pure water to remove fine particles, and then the top of the tube was filled with L-lysine solution. The tube was kept on a rotary table at room temperature for 4 days. After removing the L-lysine solution, the zeolite was washed three times in pure water and dried in an oven at 65°C for 24 hours. The nitrogen content was determined using a DeltaV isotope ratio mass spectrometer and a Flash EA 2000 elemental analyzer (both provided by Thermo Fisher Scientific).

[0107] Example 2: Nitrogen extraction from loaded zeolite As described above, basic L-amino acids are strongly adsorbed onto zeolite. To demonstrate that nitrogen release from the zeolite loaded according to the present invention is very slow in the absence of plants, a series of extraction experiments were conducted. This example was carried out according to Example 1 using zeolite loaded with (a) basic amino acids and zeolite loaded with (b) ammonium.

[0108] Example 2(a) Zeolites loaded with L-arginine or L-lysine were immersed in an extraction solvent consisting of water, 0.5 mM CaCl2 (pH 5.8), and 0.5 mM oxalic acid (pH 1.6), respectively.

[0109] Add 1 g of arginine-zeolite or lysine-zeolite and 10 mL of their respective extraction solvents to a 15 mL polypropylene test tube.

[0110] In summary, this example demonstrates that arginine strongly binds to zeolite when washed with water (H2O), calcium chloride, and oxalic acid. (See also...) Figure 10 .

[0111] Examples 2(b) and (c) Parallel experiments were conducted using ammonium-containing zeolites, compared with amino acid zeolites and commercial slow-release fertilizers containing nitrates and ammonium (Osmocote). TM The method used by The Scotts Miracle-Gro Company is similar. Ammonium-loaded zeolite was prepared by washing natural clinoptilolite-type zeolite obtained from Incal Mineral (Izmir, Turkey) with water to remove impurities. The zeolite particles were then immersed in 2500 mL of 0.2 M ammonium sulfate solution. The zeolite was placed on a rotary table at room temperature (20°C) for 3 days. The zeolite was then rinsed three times with pure water to remove excess ammonium sulfate from the zeolite surface. All test tubes were placed on a rotary table at room temperature (20°C). Samples were taken every three or four days to measure the concentrations of amino acids (a), ammonium (b), and nitrates (c). After each sampling, the solution in the tube was replaced with fresh extraction solution.

[0112] Example 2(d) In another parallel experiment, 20 ml of zeolite loaded with arginine (equivalent to 1% nitrogen) was mixed with 80 ml of soil, peat, or sand and placed in a container watered twice daily. These containers contained no plants. After three months of watering, the soil, peat, or sand was washed off, and the nitrogen content of the cleaned zeolite was analyzed. Surprisingly, most of the intact arginine remained in the zeolite, providing evidence of sustained release obtainable according to the present invention. As those skilled in the art will understand, the choice of growth medium can be used as one of the parameters that can affect the release rate of a fertilizer composition, thus providing flexibility in terms of properties.

[0113] Example 3: Greenhouse experiment using barley and lettuce Natural zeolite of the clinoptilolite type obtained from Incal Mineral (Izmir, Turkey) loaded with arginine (2% nitrogen) was mixed with unfertilized calcareous soil (Hasselfors Garden) in 80 ml pots at a concentration of 20 mg N per pot (n = 20). One seed of barley (Hordeum vulgare) and lettuce (Lactuca sativa) was sown per pot, watered, and covered with nonwoven fabric until germination. As a control, unfertilized soil without added arginine-zeolite was used. After 8 weeks, the barley and lettuce were harvested and rinsed to remove all soil from the roots. The plants were dried at 65°C for 24 h and then ground into a fine powder using a mortar and pistils. Total dry biomass was measured. Total nitrogen content was measured using a carbon / nitrogen analysis method (see above), see [link to relevant documentation]. Figures 1 to 3 The results in.

[0114] Example 4: Greenhouse Experiment Using European Red Pine Seedlings A. Natural zeolite of the clinoptilolite type (obtained from Incal Mineral (Izmir, Turkey)) loaded with arginine (2% nitrogen) was mixed with sand in 80 mL pots to a concentration of 20 mg N per pot (n = 20). One Scots pine seed was sown in each pot and watered. Plants without arginine-loaded zeolite were used as a control. After 12 weeks, Scots pine seedlings were harvested and rinsed to remove all soil from the roots. The plants were dried at 65°C for 24 hours and ground into a fine powder using a mortar and pestle and pistils. Total biomass was measured, and the results are shown in Figure 4.

[0115] B. Natural zeolite of the clinoptilolite type (obtained from Incal Mineral (Izmir, Turkey)) loaded with arginine (2% nitrogen) was mixed with sand in 80 mL pots to a concentration of 20 mg N per pot (n = 20). One European red pine seed was sown in each pot and watered. After 4 weeks, mycorrhizae were added to the seedlings. Plants without mycorrhizae were used as a control. After 12 weeks, the pine seedlings were harvested and rinsed to remove all soil from the roots. The plants were dried at 65°C for 24 hours and ground into a fine powder using a mortar and pestle. Total biomass and total nitrogen, as well as the effect of mycorrhizae, were measured, and the results are shown in [Figure number missing]. Figure 8 And 9.

[0116] Example 5: Field trial using European red pine seedlings Prior to planting, European red pine seedlings (Pinus Sylvestris) (n = 50) cultured in a conifer nursery according to standard methods were treated with zeolite loaded with arginine (2% nitrogen) to a concentration of 40 mg N per plant. The seedlings were planted in clearly demarcated areas according to standard methods for pine forests. The seedlings were planted side-by-side with untreated pine seedlings in mineral soil. After one growing season (3 months), the plants were harvested and washed in water. After drying at 65°C for 24 hours, the dry weight of the plants was measured; see Figures 4 and 5 for the results.

[0117] Example 6: Germination test of European red pine seedlings treated with zeolite loaded with arginine Zeolite loaded with arginine (2% nitrogen) was mixed with unfertilized lime peat (Hasselfors garden) in 80 mL pots to a concentration of 40 mg N per pot (n = 50), compared to a commercially available amino acid-based non-zeolite fertilizer with 40 mg N mixed into the peat. One seed of Scots pine (Pinus Sylvestris) was sown in each pot and watered. Germination rate was scored after 4 weeks. Results are as follows: Figure 6 As shown.

[0118] Example 7: Treatment of Norway spruce (Picea abies) and twisted pine (Pinus truncatum) with arginine zeolite Pine (Pinus contorta) Seedlings of Scots pine, Norway spruce, and twisted pine were cultured in a nursery according to standard methods and then planted in mineral soil. Half of the seedlings received no additional fertilizer, while the other half were fed zeolite loaded with arginine, prepared according to Example 1, to the roots of the seedlings before planting. The total amount of nitrogen added to each seedling was approximately 28 mg N. Seedlings were harvested after one growing season, and the dry biomass of branches, roots, and total biomass was recorded. The biomass of roots that appeared during the growing season (“field roots”) was also measured.

[0119] Surprisingly, it was found that zeolite loaded with arginine, according to the present invention and supplied to the roots of seedlings at planting time, had a positive effect on root biomass (Fig. 12) and growth in the field. Figure 13 It has a positive long-term impact.

[0120] Example 8: Reaction of fairway turf to zeolite loaded with L-arginine Zeolite fertilizer loaded with L-arginine showed an enhanced turf colonization rate from the seeds, indicating that amino acid-loaded zeolite can support the effective growth of grass species.

[0121] Under standard greenhouse conditions, artificial light is used to supplement 16 hours of daytime (20-25°C) and 8 hours of nighttime (at 15°C). A grass seed mixture of 70% *Festuca rubra* spp. and 30% *Poa pratensis* (known as "fairway turf"), typically used on golf fairways in temperate and cold climates, is applied at a rate equivalent to 3 kg seeds / 100 m². 2 The seeds were sown in 3L pots containing sand with approximately 10% organic matter. To support full turf cover in all pots, a 0.15 kg N / 100 m² sowing rate was applied six weeks after sowing. 2 Liquid NH4NO3 fertilizer was applied at a rate of [missing information]. Subsequently, prior to the start of the experimental period, a cut-regeneration cycle was performed four times per week, during which the grass was cut to 20 mm and debris was removed.

[0122] With the equivalent of 0.5 kg N / 100 m 2 A single treatment with zeolite loaded with arginine was performed at a rate of [missing information]. A reference treatment matched to the total nitrogen level was applied using a commercially available ammonium / urea-based product formulated for golf fairway coating, a commercially available methylurea-based product formulated for uncoated golf fairways, or chemically pure N-methylurea. A blank control was also established, which did not receive granular fertilizer treatment during the experiment. Treatments were repeated four times.

[0123] Table 1: Fertilizer Treatment

[0124] * Impact CGF is a commercial fertilizer sold by Indigrow (UK) Ltd.

[0125] **Premium elite is a commercial fertilizer sold by Skåne frö AB in Sweden.

[0126] Grass fragments were collected 20 mm above the sandy growing medium and oven-dried at 50°C once before fertilization (week 0), and then weekly for the following six weeks (weeks 1–6). Seven weeks after granular fertilizer treatment, roots were washed and oven-dried (at 50°C). The results of this example indicate that fairway turf exhibited a general increase in aboveground biomass production in response to granular nitrogen addition. Except for zeolite loaded with arginine, which showed a significant increase in biomass production in the first week after treatment, a significant increase was first observed in the second cut-regeneration cycle for all other fertilizer treatments. For all N treatments, peak biomass production levels were typically reached in the second or third cut-regeneration cycle. In the sixth cycle, aboveground biomass production in the cut-regeneration cycle decreased to below pretreatment levels for all fertilizer treatments.

[0127] In the response of uncoated N-methylurea, a leaf burn stress response, commonly referred to as “coking,” was observed, but it did not respond to zeolite loaded with arginine. N recovery was assessed by measuring N content using an elemental analyzer (Flash EA 2000, Thermo Fisher Scientific, Bremen, Germany) and adjusting for fragment biomass. Figure 14 After the initial delay, except for zeolite loaded with L-arginine which showed the highest recovery rate 3 weeks after application, N was recovered at the highest rate 2 weeks after N addition for all N treatments.

Claims

1. A method for promoting the growth of at least one plant, the method comprising: a) Provide a fertilizer composition comprising at least one zeolite having at least one basic L-amino acid adsorbed in its pores, the zeolite being washed after adsorbing at least one basic L-amino acid. b) Add the fertilizer composition to the plant growth medium related to planting; c) Provide released nitrogen from the fertilizer composition during subsequent cultivation of the plant. The amount of nitrogen derived from the basic L-amino acids in the zeolite is 1-3% based on the total weight of the zeolite, and the release provides long-term and continuous nutrition to the plant.

2. The method according to claim 1, wherein the zeolite is prepared by the following steps: i) Prepare an aqueous solution of alkaline L-amino acids and adjust the pH to 3-9; ii) Rinse the zeolite in water to remove dust particles and other impurities; iii) Add an aqueous solution of alkaline L-amino acids to the rinsed zeolite and stir the mixture at room temperature for 2-4 days; iv) Thoroughly wash the zeolite in water; and v) Dry the washed zeolite.

3. The method according to claim 1 or 2, wherein the basic L-amino acid is selected from L-arginine and L-lysine.

4. The method according to claim 1 or 2, wherein one or more additional growth-promoting components are added.

5. The method according to claim 1 or 2, wherein the zeolite is a natural zeolite.

6. The method according to claim 1 or 2, wherein the plant is a tree.

7. The method of claim 6, wherein the tree is a coniferous tree.

8. The method of claim 6, wherein the tree is a member of the pine family.

9. The method of claim 6, wherein the tree is a member of the genus *Pinus* or *Picea*.

10. The method according to claim 1 or 2, wherein the plant is a grass.

11. The method of claim 10, wherein the grass is a member of the Poaceae family.

12. A growth support material comprising a medium for plant growth, comprising at least one zeolite having at least one basic L-amino acid adsorbed in its pores, optionally together with other growth-promoting components, wherein the loading of nitrogen derived from the basic L-amino acid in the zeolite is 1-3% based on the total weight of the loaded zeolite, the zeolite is washed after adsorbing the at least one basic L-amino acid, and the zeolite provides nitrogen release to provide long-term and continuous nutrition to the plant. Zeolite is prepared through the following steps: i) Prepare an aqueous solution of alkaline L-amino acids and adjust the pH to 3-9; ii) Rinse the zeolite in water to remove dust particles and other impurities; iii) Add an aqueous solution of alkaline L-amino acids to the rinsed zeolite and stir the mixture at room temperature for 2-4 days; iv) Thoroughly wash the zeolite in water; and v) Dry the washed zeolite.

13. The growth support material of claim 12, wherein it is provided in a biodegradable container.

14. The growth support material according to claim 12, wherein it is a pad for seed planting.

15. Use of at least one zeolite having at least one basic L-amino acid adsorbed in its pores, optionally as a fertilizer along with other growth-promoting components, wherein the loading of nitrogen derived from said basic L-amino acid in the zeolite is 1-3% based on the total weight of the loaded zeolite, the zeolite is washed after adsorbing said at least one basic L-amino acid, and the zeolite provides nitrogen release to provide long-term and continuous nutrition to the plant. Zeolite is prepared through the following steps: i) Prepare an aqueous solution of alkaline L-amino acids and adjust the pH to 3-9; ii) Rinse the zeolite in water to remove dust particles and other impurities; iii) Add an aqueous solution of alkaline L-amino acids to the rinsed zeolite and stir the mixture at room temperature for 2-4 days; iv) Thoroughly wash the zeolite in water; and v) Dry the washed zeolite.

16. The use according to claim 15, wherein the growth support material of any one of claims 12-14 is combined with at least one plant.

17. The use according to claim 15 or 16, wherein the loading of nitrogen derived from the basic L-amino acid in the zeolite is at least 2% based on the total weight of the loaded zeolite.

18. The use according to claim 16, wherein the plant is a conifer.

19. The use according to claim 16, wherein the plant is a member of the Pinaceae family.

20. The use according to claim 16, wherein the plant is a member of the genus *Pinus* or *Picea*.

21. The use according to claim 16, wherein the plant is a grass.

22. The use according to claim 16, wherein the plant is a member of the Poaceae family.

23. The use according to claim 15 or 16, wherein it is used for cultivating at least one mycorrhizal plant.

24. The use according to claim 16, wherein the plant is capable of symbiotic relations with the fungus.

25. The use according to claim 24, wherein the fungus produces at least one edible mushroom.

26. The use according to claim 25, wherein the edible mushroom is chanterelle.

Images