Use of denatonium benzoate as a crop seed treatment agent as a bird and / or insect repellent

Abstract

The present invention relates to the protection of seeds and plants from damage caused by birds and / or insects. Treatment with denatonium benzoate provides seeds protected from attack by birds, especially corvids, and insects, especially nematodes.

Description

[0001] The present invention relates to protecting seeds and plants from damage caused by birds and / or insects. Treatment with denatonium benzoate provides seeds protected from attack by birds, especially corvids, and insects, especially wireworms, grain beetles or cabbage stem flea beetles.

[0002] Birds of the genus Corvus, commonly called corvids, are widespread and abundant throughout Europe and North America throughout the year. The diet of corvids includes insects, worms, snails, slugs, berries, legumes, and cereals. The worst damage to cereals by corvids is caused by feeding on seeds before or after emergence, resulting in a reduction in plant populations in the field. Maize (corn) is severely damaged by corvid feeding activities. Corvids are very efficient at locating seeds in newly sown crops, as shown by the pattern of "dig" holes where the damage occurs. In newly emerged crops, corvids will uproot seedlings that are attached to the seeds with some endosperm remaining.

[0003] Today, in maize cultivation, there are two main substances used for bird control: Mesurol, which contains the active ingredient methiocarb (4 (methylthio) 3,5-xylyl-N-methylcarbamate), and Thiram (tetramethylthiuram sulphide). These substances have been used for decades, and many studies have been conducted on their efficiency as bird repellents, and many studies have shown that these substances have strong effects as bird repellents without affecting the germination ability of seeds. However, there are also studies showing that, for example, Thiram seems to significantly reduce early seedling vigor (Sandhu et al. (1987). Evaluation of methiocarb and thiram as seed treatments for protecting sprouting maize from birds in Punjab (India). International Journal of Pest Management, 33 (4), 370-372.). Since the European Commission has recently banned several chemicals, alternative substances are needed.

[0004] Another pest of seeds is insects and insect larvae, which also often cause damage by feeding on seeds. An example of such insect pests is nematodes. Nematodes are larvae of click beetles (Coleoptera: Elateridae). They consist of more than 9,000 species distributed throughout the world, some of which are important pests of various crops such as corn, potatoes, cereals, carrots, sugar beets, sugar cane and soft fruits. In Europe, the damage caused by nematode infestation is mainly attributed to the genus Agriotes Eschscholtz. It contains more than 200 species worldwide, including more than 100 species in the Palearctic region. The Agriotes community is widely distributed throughout the continent. The genus Melanotus Eschscholtz has been studied in depth in Japan (Melanotus okinawensis Ohira) and the United States (Melanotus communis (Gyll.)). The "Pacific coast" nematode, Limoniuscanus Leconte, along with other species of this genus, causes damage to crops along the west coast of North America. Other genera of importance to agriculture are Athous Eschscholtz, Conoderus Eschscholtz, Ctenicera Latreille, and Hypolithus Eschscholtz.

[0005] The main effect of nematode feeding on stems and underground plant organs is seedling mortality and all the yield losses that implies. Nematode larvae may feed on the germ of a corn kernel or completely hollow out the seed, leaving only the seed coat. This damage can cause gaps to appear in the rows. Nematodes can also cut rootlets or tunnels into the roots or underground parts of the stem of young corn plants. These plants will appear stunted or wilted, with the whorl leaves wilting first.

[0006] Nematode damage occurs primarily during the early stages of plant growth. Damage typically occurs when corn is planted early and the weather cools enough to slow seed germination. Nematode infestations are typically found in areas of the field that remain wet for long periods of time.

[0007] Nematode management in corn is typically performed through seed treatments or treatments at planting. Cultural practices such as crop rotation, flooding, or cultivation are generally ineffective because nematodes have a broad host range and can adapt to different environments. Delaying planting to allow soil drying and warming can improve corn emergence because nematodes retreat deep into the soil under these conditions, but this management practice may not always be possible.

[0008] Another important insect pest is the grain beetle (Zabrus tenebrioides), which damages all cereal crops, especially early varieties (winter varieties). The larvae and beetles especially damage the leaf tissue of cereal crops. In oilseed rape, the cabbage stem flea beetle (Psylliodes chrysocephala) is a problem. The lack of effective chemical options in oilseed rape has accelerated the development of cabbage stem flea beetles. Currently, this beetle is the number one pest of this crop and can damage the growing points of the plants and cause crop failure.

[0009] A disadvantage of many insecticides commonly used to protect seeds is that they are also dangerous to non-target insects, especially bees. Information from standard tests and field studies as well as incident reports involving neonicotinoid insecticides (e.g., clothianidin) indicate that there may be a long-term toxicity risk to bees and other beneficial insects.

[0010] Therefore, the problem underlying the present invention is to provide a seed treatment agent suitable for protecting corn seeds from damage caused by birds and / or insects, preferably without impairing the germination ability of the seeds. Compromising the germination and quality of the seeds may result in a significant reduction in yield. In this context, it is particularly desirable to have a substance that can protect corn seeds and seedlings, but which is safe for plant consumption and preferably detectable for a longer period of time in mature plants.

[0011] A substance often suggested for use as an animal repellent is denatonium benzoate (N-benzyl-2-(2,6-dimethylanilino)-N,N-diethyl-2-oxoethane-1-aminium benzoate, Bitrex), which is a bittering agent used to prevent, for example, accidental poisoning of children (Sibert & Frude (1991). Emergency Medicine Journal, 8 (1), 1-7.). However, there are few studies on the use of denatonium benzoate as a bird repellent, especially a corvid bird repellent, in cereal plants. Kennedy and Connery ((2008). An investigation of seed treatments for the control of crow damage to newly-sown wheat. Irish Journal of Agricultural and Food Research, 79-91.) reported on the use of different substances as corvid bird repellents in wheat. They concluded that denatonium benzoate provided only a modest improvement, and at certain concentrations, the damage caused by crows was even higher than that of untreated controls.

[0012] However, in the present invention, it is surprisingly found that, especially for seeds, especially for corn seeds, denatonium benzoate provides excellent protection to prevent bird damage. The activity of denatonium benzoate as a Corvidae bird repellent for example corn is similar to or even better than the best bird repellents such as Thiram, Korit and Mesurol. Although previous studies on the use of denatonium benzoate as a wheat seed treatment agent have concluded that denatonium benzoate cannot provide protection to prevent Corvidae birds, it is surprisingly found that for example corn, denatonium benzoate is particularly effective. In addition, it has been found that denatonium benzoate also provides excellent protection for seeds of crops such as corn, rape or rye to prevent damage caused by insects and insect larvae, particularly to prevent nematodes, grain beetles or cabbage stem flea beetles. The activity of denatonium benzoate as an insect repellent for corn, oilseed rape or rye, preferably as a nematode, grain beetle or cabbage stem flea beetle repellent, is acceptable and reaches levels similar to standard insecticides such as tefluthrin and clothianidin. In addition, it is advantageous because it is harmless to beneficial insects.

[0013] In addition, denatonium benzoate has been found to have a systemic effect on crop plants such as corn, rapeseed and rye. This protects not only the seeds, but also the sprouts that emerge. Since denatonium benzoate is diluted during growth and maturation, the harvested crops (e.g. corn cobs, rapeseed or rye grains) do not show an effect on the amount of denatonium that is used after the harvest of these crops (e.g. for animal feeding or silage purposes). In addition, it has been found that treatment with denatonium benzoate is safe for seeds and does not reduce the germination or quality of the seeds.

[0014] Therefore, in a first aspect, the present invention provides a plant seed treated with denatonium methyl, wherein the denatonium methyl is adsorbed on the seed and / or absorbed into the seed. Therefore, the denatonium methyl may be present on the surface of the seed, inside the seed, or at both locations. If the denatonium methyl is present on the surface of the seed, the wording "seed coated with denatonium methyl" may be used alternatively in this application. However, the term "coating" does not mean that the seed is completely covered with denatonium methyl, but refers to that the denatonium methyl adheres to at least a portion of the seed surface.

[0015] According to a preferred embodiment of the present invention, denatonium benzoate is at least partially absorbed into the seed. Thus, denatonium benzoate is at least partially absorbed by the seed, but may also be partially retained outside the seed. Absorption of denatonium benzoate into the seed prevents the active agent from being washed off the seed surface, for example by rain or mechanically rubbed off, and allows the bird and / or insect repellent activity to be maintained.

[0016] According to the present invention, denatonium benzoate can be used in the form of a single active agent or optionally in combination with one or more further active agents and / or additives. The one or more active agents that can be used can be any preparation, preferably a chemical preparation that produces a desired effect on seeds, plants that eventually grow from the seeds, or both. Non-limiting examples of such chemical preparations include pesticides (such as fungicides, miticides, acaricides, insecticides, insect repellents, rodenticides, molluscicides, nematicides, bactericides, and fumigants), herbicides, chemical hybrids, plant growth hormones, antibiotics and other drugs, bioattractants, growth regulators, pheromones, and dyes. Specific non-limiting examples of chemical preparations that can be used as active ingredients include triticonazole, imidacloprid, tefluthrin, and silthiophenamide (silthiophenamide, N-allyl-4,5-dimethyl-2-trimethylsilylthiophene-3-carboxamide). Original textcaboxamide (carboxamide) is suitable for further active agents of the present invention, for example, phytochemicals and antimicrobials suitable for protecting plant seeds. Preferred examples thereof are bactericides, antiparasitics and fungicides, such as DSM RD9103BRP, pepper extract, black cumin oil, yucca extract, mentha pulegium extract, ammonium aluminum sulfate, hop extract, fat-encapsulated plant extracts, alginate-encapsulated plant extracts, tefluthrin, yellow substances, metalaxyl, fludioxonil, prothioconazole and combinations thereof, preferably Redigo M and / or Maxim XL.

[0017] Potential additives may be adhesives that improve the adhesion of denatonium benzoate to plant seeds. Particularly preferred are non-hazardous adhesives, preferably adhesives based on natural resins, such as Satec WFO20, COMBICOAT or Satec C, adhesives based on lignin sulfonates, such as Satec WWL, adhesives based on polymers, such as Satec RHI, and / or water-based adhesives, such as Quality or Active, Disco L-450; Disco L-1101, Agifix, etc. Further exemplary additives are fillers or extenders and binders. In addition, talc or graphite powder can be added to prevent the seeds from agglomerating.

[0018] As used herein, the term "corn" means a plant of the species Zea mays. The term corn is interchangeable with maize and maize. As used herein, the term "rapeseed" or "oilseed rape" means a plant of the species Brassicanapus. As used herein, the term "rye" means a plant of the species Secale cereale.

[0019] Plant seeds according to the present invention preferably contain an effective amount of denatonium benzoate to repel birds and / or insects and insect larvae, and thus provide protection to prevent damage caused by birds and / or insects. Specifically, the amount of denatonium benzoate on and / or in the seeds according to the present invention can effectively repel birds of the genus Corvus, which are commonly known as corvids, such as crows, ravens, rooks and jackdaws. Birds are prevented from eating seeds. However, their health is not affected. Further, insects and insect larvae such as nematodes, grain beetles or cabbage stem flea beetles are repelled, but are not affected in terms of health. Denatonium benzoate is also harmless to the health of other animals that may come into contact with the seeds according to the present invention.

[0020] For the purposes of the present invention, the term "insect" shall in principle cover all types of insects at any stage of development, including adults and larvae. Advantageously, harmful insects such as nematodes, grain beetles or cabbage stem flea beetles are repelled and thus prevented from feeding on seeds or developing plant parts. Thus, preferably, the term insect means nematodes, grain beetles (Zabrus tenebrioides) or cabbage stem flea beetles (Psylliodes chrysocephala).

[0021] According to the present invention, nematodes mean the larvae of the click beetle belonging to the family of click beetles. Other names of click beetles include elater, snapping beetle, spring beetle or skipjack. There are about 9300 known species in the world, and 965 effective species in North America. Nematodes as used herein are agricultural pests. The larvae are slender, cylindrical or slightly flat, and the body is hard. The three pairs of legs on the thoracic segment are shorter, and the last abdominal segment is the same as the situation in the beetle larvae, usually pointing downward, and can be used as the terminal forelimbs in some species. The ninth segment (i.e. the last segment) is pointed in the larvae of the genus Melanonotus, Dalopius and the genus Melanonotus, but in the species of Selatosomus (formerly known as Ctenicera), Limonius, Hypnoides and Athous, it is bifid due to the so-called tail notch. The back of the ninth abdominal segment may also have a sharp protrusion, as in Oestodini, including the genera Drapetes and Oestodes. Although some species complete their development in one year (e.g., Conoderus), most nematodes spend three or four years in the soil, feeding on the roots of rotten vegetation and plants, and often causing damage to crops such as potatoes, strawberries, corn and wheat. According to the present invention, the term nematode may be understood as the larvae of the click beetle belonging to the genus selected from the group consisting of the following: click beetle, Dalopius, comb claw click beetle, bright click beetle, Limonius, Hypnoides, Hypolithus, Athous, Drapetes, Oestodes or wide chest click beetle.

[0022] A further advantage of the seeds according to the invention which have been treated with denatonium benzoate is that the germination capacity of the seeds is similar to that of untreated seeds of the same species or of the same genotype. Healthy growing seedlings and plants develop from the seeds according to the invention to preferably at least the same extent as from untreated seeds. A similar level of germination capacity means that the treated seeds of a plant variety show a germination rate which deviates by a maximum of + / -10% from the germination rate of untreated seeds of the same plant variety, preferably a germination rate which deviates by a maximum of + / -5%, more preferably a germination rate which deviates by a maximum of + / -3% from the germination rate of untreated seeds of the same plant variety.

[0023] As used herein, the seed, plant or part thereof may be derived from any plant species, preferably, the seed, plant or part thereof belongs to a species selected from the group consisting of carrot (Daucus carota), cucumber (Cucumis sativus), Cucurbita argyrosperma, black seed pumpkin (Cucurbita ficifolia), Indian pumpkin (Cucurbita maxima), Chinese pumpkin (Cucurbita moschata), watermelon (Citrullus lanatus), American squash (Cucurbita pepo), strawberry (Fragaria×ananassa), rapeseed (Brassica napus), cabbage (Brassica oleracea), onion (Allium cepa), sugar beet, Swiss chard or red beet (Betavulgaris), beans (Phaseolus vulgaris), soybeans (Glycine max), peas (Pisum sativum), peppers (Capsicum spp.), spinach (Spinacia oleracea), celery (Apium More preferably, the seed, plant or part thereof belongs to a species selected from the group consisting of rapeseed (Brassica napus), cabbage (Brassica oleracea), sorghum (Sorghum bicolor), corn (Zea mays), barley (Hordeum vulgare) or rye (Secale cereale).

[0024] A further aspect of the present invention is a plant or part thereof grown from a seed according to the present invention. Since denatonium benzoate surprisingly has a systemic effect and can be at least partially absorbed into the seed, it is still present in the grown seedling. The concentration in the seedling is generally lower than the concentration in the seed in which the seedling is grown. Since denatonium benzoate is diluted during growth and maturation, the crop (e.g., cob, leaf, root, tuber or any fruit) harvested does not show an amount of denatonium that affects post-harvest use (e.g., for animal feeding, food / feed production, silage purposes or human consumption).

[0025] In plants developed from seeds according to the invention, denatonium benzoate showed concentration differences between different plant parts / organs and developmental stages. In the early stages, denatonium benzoate was selectively detectable. The concentrations in the whole plant and / or leaf tissue and root tissue were generally in the ppm range, and the concentrations in the buds were in the higher ppb range. In middle-aged plants, the concentrations in the leaves were generally in the lower ppb range. The variation in concentrations varied between types (between) and plants (within), and the concentration of denatonium benzoate decreased significantly from the early stages to the mid-stages.

[0026] In a specific embodiment of the present invention, the plant is a seedling and contains an average concentration of 1 to 200 ppm or 1 to 100 ppm, preferably 1 to 50 ppm, more preferably 5 to 30 ppm, even more preferably 10 to 22 ppm of denatonium benzoate throughout the seedling. A further embodiment is a seedling containing an average concentration of 50 to 4000 ppb, preferably 50 to 2500 ppb, more preferably 100 to 1500 ppb, even more preferably 200 to 800 ppb of denatonium benzoate in the shoot tissue of the seedling. Another embodiment is a seedling containing an average concentration of 5 to 50,000 ppb, preferably 5 to 25,000 ppb, more preferably 5 to 10,000 ppb, even more preferably 5 to 5,000 ppb or 5 to 2500 ppb or 5 to 1000 ppb of denatonium benzoate in the leaf tissue of the seedling. A further embodiment is a plant at development stage V6-V8, containing in the leaf tissue of the plant an average concentration of 0.5 to 200 ppb, preferably 0.5 to 100 ppb, more preferably 1 to 50 ppb, even more preferably 5 to 20 ppb of denatonium benzoate. The amount of denatonium benzoate in the developing plant is still sufficient to repel birds, especially birds of the corvidae family, and / or insects, especially nematodes, grain beetles or cabbage stem flea beetles. In further embodiments, the plant is a seedling and contains an average concentration of at least 1 ppm, at least 2 ppm, at least 3 ppm, at least 5 ppm, at least 10 ppm, at least 20 ppm, at least 30 ppm, at least 50 ppm, at least 100 ppm or at least 200 ppm of denatonium benzoate throughout the seedling or an average concentration of at least 50 ppb, at least 75 ppb, at least 100 ppb, at least 150 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 750 ppb, at least 1000 ppb in the shoot tissue of the seedling. ppb, at least 2000ppb or at least 4000ppb of denatonium benzoate, or the seedling contains an average concentration of at least 5ppb, at least 10ppb, at least 20ppb, at least 30ppb, at least 50ppb, at least 100ppb, at least 200ppb, at least 300ppb, at least 500ppb, at least 1000ppb, at least 2000ppb, at least 3000ppb, at least 5000ppb, at least 10000ppb, at least 20000ppb, at least 30000ppb or at least 50000ppb of denatonium benzoate in leaf tissue of the seedling.

[0027] In one embodiment of the invention, the plant is a corn seedling and contains an average concentration of 1 to 200 ppm or 1 to 100 ppm, preferably 1 to 50 ppm, more preferably 5 to 30 ppm, even more preferably 10 to 22 ppm of denatonium benzoate in the entire seedling. A further embodiment is a corn seedling containing an average concentration of 50 to 4000 ppb, preferably 50 to 2500 ppb, more preferably 100 to 1500 ppb, even more preferably 200 to 800 ppb of denatonium benzoate in the shoot tissue of the seedling. Another embodiment is a corn seedling containing an average concentration of 1 to 2000 ppb, preferably 1 to 1500 ppb, more preferably 1 to 1000 ppb, even more preferably 1 to 500 ppb of denatonium benzoate in the leaf tissue of the seedling. A further embodiment is a corn plant at development stage V6-V8, containing an average concentration of 0.5 to 200 ppb, preferably 0.5 to 100 ppb, more preferably 1 to 50 ppb, even more preferably 5 to 20 ppb of denatonium benzoate in the leaf tissue of the plant. The amount of denatonium benzoate in the developing plant is still sufficient to repel birds, especially corvids, and / or insects, especially nematodes. In a further embodiment, the plant is a corn seedling and contains an average concentration of at least 1 ppm, at least 2 ppm, at least 3 ppm, at least 5 ppm, at least 10 ppm, at least 20 ppm, at least 30 ppm, at least 50 ppm, at least 100 ppm, at least 150 ppm or at least 200 ppm of denatonium benzoate throughout the seedling, or an average concentration of at least 50 ppb, at least 75 ppb, at least 100 ppb, at least 150 ppb, at least 200 ppb, at least 300 ppb in the shoot tissue of the seedling. b, at least 500 ppb, at least 750 ppb, at least 1000 ppb, at least 2000 ppb or at least 4000 ppb of denatonium benzoate, or the seedlings contain an average concentration of at least 1 ppb, at least 2 ppb, at least 3 ppb, at least 5 ppb, at least 10 ppb, at least 20 ppb, at least 30 ppb, at least 50 ppb, at least 100 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 1000 ppb or at least 2000 ppb of denatonium benzoate in leaf tissue.

[0028] In another embodiment of the invention, the plant is a rapeseed seedling and contains an average concentration of 10 to 20,000 ppb, preferably 10 to 15,000 ppb or 10 to 10,000 ppb, more preferably 20 to 7,500 ppb, even more preferably 20 to 5,000 ppb of denatonium benzoate in the leaf tissue of the seedling. The amount of denatonium benzoate in the developing plant is still sufficient to repel birds, especially birds of the corvidae family, and / or insects, especially cabbage stem flea beetles. In a further embodiment, the plant is a rapeseed seedling and contains an average concentration of at least 10 ppb, at least 20 ppb, at least 30 ppb, at least 50 ppb, at least 100 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 1000 ppb, at least 2000 ppb, at least 3000 ppb, at least 5000 ppb, at least 10000 ppb, at least 15000 ppb or at least 20000 ppb of denatonium benzoate in leaf tissue of the seedling.

[0029] In a further embodiment of the invention, the plant is a rye seedling and contains an average concentration of 1 to 200 ppm or 1 to 100 ppm, preferably 1 to 50 ppm, more preferably 5 to 30 ppm, even more preferably 10 to 22 ppm of denatonium benzoate in the leaf tissue and / or root tissue of the seedling. The amount of denatonium benzoate in the developing plant is still sufficient to repel birds, especially corvids, and / or insects, especially grain beetles. In a further embodiment, the plant is a rye seedling and contains an average concentration of at least 1 ppm, at least 2 ppm, at least 3 ppm, at least 5 ppm, at least 10 ppm, at least 20 ppm, at least 30 ppm, at least 50 ppm, at least 75 ppm, at least 100 ppm, at least 150 ppm or at least 200 ppm of denatonium benzoate in the leaf tissue and / or root tissue of the seedling.

[0030] As used in this application, the term "seedling" refers to a plant from emergence to the three-leaf stage of a monocot or the second pair of secondary leaves, preferably the first pair of secondary leaves, of a dicot. Preferably, the seedlings of monocots such as maize or Secale graine are in a developmental stage from emergence (i.e., the coleoptile penetrates the soil surface (cracking stage)) to the V3 developmental stage (i.e., three leaves unfolded), more preferably the V2 developmental stage (i.e., two leaves unfolded).

[0031] As used herein, the middle developmental stage of a plant means the developmental stage after germination and seed emergence and the seedling stage but before maturity and harvest. Preferably, at this stage, the plant contains six to eight fully grown leaves. More preferably, the reproductive stage of the plant begins. The reproductive stage of plant development occurs after the juvenile stage is completed. At this stage, the plants are considered mature, that is, they are physiologically able to begin to produce reproductive parts: flowers, fruits and seeds. This stage consists of the period from when the plant begins to form inflorescences or flower primordia until flowering, pollination and fertilization.

[0032] In another specific embodiment of the invention, the plant is a seedling and contains an average amount of 1 to 100 μg per plant, preferably 1 to 40 μg per plant, more preferably 2 to 25 μg per plant, even more preferably 5 to 15 μg of denatonium benzoate per whole plant. A further embodiment is a seedling containing denatonium benzoate in an average amount of 0.01 to 1 μg per seedling sprout, preferably 0.01 to 0.5 μg per seedling sprout, more preferably 0.03 to 0.3 μg per seedling sprout, even more preferably 0.05 to 0.2 μg per seedling sprout. In developing plants, such as seedlings, the amount of denatonium benzoate is still sufficient to repel birds, especially birds of the corvidae family, and / or insects, especially nematodes, grain beetles or cabbage stem flea beetles.

[0033] In addition to denatonium benzoate, the plant or its part according to the present invention may also contain one or more further active agents and / or additives in addition. These additional active agents and / or additives are preferably those mentioned above in the context of the seed. Depending on the extent to which these further active agents and / or additives are absorbed by the seed, their concentration in the plant or its part may vary. Active agents and additives that are not absorbed by the seed or are only absorbed by the seed in small amounts may still be present on the surface of the plant or its part.

[0034] Another aspect of the invention is the use of denatonium benzoate for protecting plant seeds or plants, preferably seedlings, from damage caused by birds and / or insects. For this purpose, denatonium benzoate can be optionally combined with one or more further agents and / or additives, preferably those defined above with reference to the seeds according to the invention. In principle, denatonium benzoate can be applied to seeds or plants, or can be applied together with seeds during sowing in agricultural areas. It is preferred to pre-treat the seeds, because the amount of denatonium benzoate required is significantly lower than the amount applied only during sowing.

[0035] Furthermore, the present invention relates to a method for protecting seeds from damage caused by birds and / or insects, the method comprising contacting the seeds with denatonium benzoate. Optionally, denatonium benzoate may be combined with one or more further active agents and / or additives as described above.

[0036] As used herein, "contacting" means that the seed is brought into contact with the denatonium benzoate in some manner. The contact may result in a more or less continuous coating of the seed. For the purposes of the present invention, coating does not necessarily mean that the seed surface is completely coated with the denatonium benzoate, but includes any kind of adhesion of the denatonium benzoate to any part of the seed surface. There does not necessarily have to be direct contact between the denatonium benzoate and the seed surface, but in principle adhesion may also occur via any intermediate layer or substance.

[0037] Any method known in the art can be used to contact the seed with denatonium benzoate so as to apply the active agent to the seed. Exemplary techniques suitable for effectively applying denatonium benzoate uniformly to the seed surface and uniformly from seed to seed are granulation and film coating methods. Granulation is defined as the deposition of a layer of inert material that may mask the original shape and size of the seed, thereby causing a significant increase in weight and improving seed suitability. Film coating retains the shape and general size of the original seed with minimal weight gain.

[0038] Seed granulation is composed of the following methods: solid particles acting as fillers (e.g., diatomaceous earth, calcium carbonate or silicon dioxide) or fillers are applied together with adhesives or tackifiers to form more or less spherical dispersed units. The granulation process is usually carried out in a coating pan or drum. The adhesive or tackifier can be mixed with water and sprayed on the seeds, or dry mixed with fillers. The coating formulation can be a blend of adhesive or tackifier and filler, which is formulated into a dry powder. In an alternating manner, during the coating operation, the powder is sprinkled on the seeds and water is sprayed on the seeds. Water temporarily binds the pellets and activates the tackifier. Granulation is a wet operation, so the pellets must be dried when the coating process is completed.

[0039] The adhesive used for seed coating composition is well-known in the art, and its example comprises water-soluble polymer, as polyvinyl acetate, polyvinyl alcohol, polyvinyl pyrrolidone, polyurethane, methylcellulose, carboxymethyl cellulose, hydroxypropyl cellulose, sodium alginate, polyurethane, polyacrylate, casein, gelatin, amylopectin, polyacrylamide, polyethylene oxide, polystyrene, styrene acrylic acid copolymer, styrene butadiene polymer and poly-(N-vinylacetamide).In addition, waxes such as carnauba wax (carnauba wax), paraffin, polyethylene wax, beeswax and polypropylene wax also can be used as adhesive or extra flow additive.In addition, ethylene vinyl acetate also can be suitably used as adhesive.

[0040] Thin-film coating of seeds consists of spraying a solution or suspension of a film-forming polymer onto the seeds to achieve a uniform deposition of the material. Coating pans for granulation can also be used; however, in contrast to wet operations for granulation, aqueous film-forming formulations must be dried immediately after spraying to avoid agglomerations.

[0041] Both granulation and film coating can be used as a delivery system for denatonium benzoate and optional further active agents and / or additives at high loading rates. In the case of granulation, the active ingredient formulated as a dry powder can be applied in the outer granular layer. The final coating application using a filler material encapsulates the plant protection agent and prevents workers from contacting the active ingredient.

[0042] In a preferred aspect of the present invention, the amount of denatonium benzoate is 0.1 to 70 g / 50,000 corn seeds, 0.1 to 50 g / 50,000 corn seeds, 0.1 to 20 g / 50,000 corn seeds or 0.1 to 10 g / 50,000 corn seeds, particularly 0.2 to 5 g / 50,000 corn seeds. In the case where denatonium benzoate is used in the form of a single active agent to prevent bird damage, the amount of denatonium benzoate used according to the present invention is preferably 0.5 to 1.5 g / 50,000 corn seeds. When denatonium benzoate is used in combination with one or more further active agents, a lower amount of denatonium benzoate may be sufficient, preferably 0.2 to 1 g / 50,000 corn seeds. For the purpose of insect repellents, even higher concentrations can be applied to seeds, which are as high as 50 to 70 g / 50,000 corn seeds. In another embodiment, the amount of denatonium benzoate is at least 0.2 g / 50,000 corn seeds, at least 0.4 g / 50,000 corn seeds, at least 0.5 g / 50,000 corn seeds, at least 0.8 g / 50,000 corn seeds, at least 1 g / 50,000 corn seeds, at least 2 g / 50,000 corn seeds, at least 5 g / 50,000 corn seeds, at least 10 g / 50,000 corn seeds, at least 20 g / 50,000 corn seeds, at least 30 g / 50,000 corn seeds, at least 50 g / 50,000 corn seeds, or at least 70 g / 50,000 corn seeds.

[0043] In a preferred aspect of the invention, the amount of denatonium benzoate is 2 to 200 g / 1.5 million rapeseed seeds (equivalent to about 7 kg of seeds), 2 to 100 g / 1.5 million rapeseed seeds, 2 to 50 g / 1.5 million rapeseed seeds, 2 to 20 g / 1.5 million rapeseed seeds or 2 to 10 g / 1.5 million rapeseed seeds, in particular 2 to 25 g / 1.5 million rapeseed seeds. In the case where denatonium benzoate is used as a single active agent to prevent bird damage, the amount of denatonium benzoate used according to the invention is preferably 2 to 100 g / 1.5 million rapeseed seeds. When denatonium benzoate is used in combination with one or more further active agents, a lower amount of denatonium benzoate may be sufficient, preferably 2 to 50 g / 1.5 million rapeseed seeds. For the purpose of insect repellents, even higher concentrations can be applied to the seeds, which are up to 50 or even 200 g / 1.5 million rapeseed seeds. Preferably, almost all or all of the aforementioned 1.5 million rapeseed seeds are viable. In another embodiment, the amount of denatonium benzoate used is at least 2 g / 1.5 million rapeseed seeds, at least 5 g / 1.5 million rapeseed seeds, at least 10 g / 1.5 million rapeseed seeds, at least 20 g / 1.5 million rapeseed seeds, at least 50 g / 1.5 million rapeseed seeds, or at least 200 g / 1.5 million rapeseed seeds.

[0044] In another preferred aspect of the invention, the amount of denatonium benzoate used is 10 to 1500 g / 100 kg rye seeds, 10 to 1000 g / 100 kg rye seeds, 10 to 750 g / 100 kg rye seeds, 10 to 500 g / 100 kg rye seeds or 250 g / 100 kg rye seeds, in particular 10 to 75 g / 100 kg rye seeds or 10 to 150 g / 100 kg rye seeds. In the case where denatonium benzoate is used as a single active agent to prevent bird damage, the amount of denatonium benzoate used according to the invention is preferably 10 to 75 g / 100 kg rye seeds. When denatonium benzoate is used in combination with one or more further active agents, lower amounts of denatonium benzoate may be sufficient, preferably 10 to 50 g / 100 kg rye seeds. For insect repellent purposes, even higher concentrations can be applied to the seeds, up to 150 or even 1500 g / 100 kg rye seeds. In another embodiment, the amount of denatonium benzoate used is at least 10 g / 100 kg rye seeds, at least 20 g / 100 kg rye seeds, at least 30 g / 100 kg rye seeds, at least 50 g / 100 kg rye seeds, at least 75 g / 100 kg rye seeds, at least 100 g / 100 kg rye seeds, at least 150 g / 100 kg rye seeds, at least 200 g / 100 kg rye seeds, at least 300 g / 100 kg rye seeds, at least 500 g / 100 kg rye seeds, at least 750 g / 100 kg rye seeds, at least 1000 g / 100 kg rye seeds or at least 1500 g / 100 kg rye seeds.

[0045] Basically, denatonium benzoate can be administered in any form. For example, denatonium benzoate can be used in the form of a solid powder in crystalline pure form (CAS: 3734-33-6; ai=3500ppm), or in the form of a liquid composition, or in the form of a concentrated solution of at least 50% dissolved denatonium benzoate, at least 60% dissolved denatonium benzoate or even at least 70% dissolved denatonium benzoate. Preferably, the liquid composition used according to the present invention further comprises one or more adhesives. In a particular aspect of the present invention, the liquid composition comprises denatonium benzoate in a concentration of about 1-800 g / l, about 1-500 g / l, about 1-250 g / l, about 1-100 g / l, in particular 10-50 g / l, preferably 13-27 g / l. In another specific aspect of the present invention, the liquid composition comprises the denatonium benzoate in a concentration of at least 10 g / l, at least 25 g / l, at least 50 g / l, at least 100 g / l, at least 250 g / l or at least 500 g / l. In addition, the liquid composition may include one or more further activating agents or additives, preferably as described above. Alternatively or additionally, the further activating agent may be applied to the seed in the form of a separate composition. In this case, an additional activating agent may be used to process before or after processing with the denatonium benzoate, or denatonium benzoate and an additional activating agent may be used to process simultaneously.

[0046] A further aspect of the invention relates to a method for providing plants protected from damage by birds and / or insects, the method comprising

[0047] a) providing a plant seed as described above or a seed obtained or obtainable according to the method described above, and

[0048] b) growing plants from the seeds of a).

[0049] The seeds provided in step (a) do not need to be treated with denatonium benzoate immediately prior to being subjected to the method, but seeds may be used which have been treated at some earlier stage and then stored.

[0050] The conditions used for growing the plants in step b) correspond to the conditions usually applied for growing said plants or plant varieties. The fact that the seeds have been treated with denatonium benzoate does not impose any special requirements on the growing conditions. In particular, the seeds can be sown in agricultural areas and will germinate, grow and mature as long as there are sufficient nutrients, water and light available.

[0051] Since denatonium benzoate is at least partially absorbed by the seeds, the resulting plant preferably contains denatonium benzoate. This also protects the emerging seedlings and developing plants from attack by birds, in particular birds of the corvidae family, and / or insects, in particular nematodes, grain beetles or cabbage stem flea beetles and their larvae. Optionally, the plant may further contain one or more additional active agents and / or additives, in particular those described above.

[0052] The plant may, for example, be a seedling containing an average concentration of 1 to 200 ppm or 1 to 100 ppm, preferably 1 to 50 ppm, more preferably 5 to 30 ppm, even more preferably 10 to 22 ppm of denatonium benzoate in the entire seedling; or a seedling containing an average concentration of 50 to 4000 ppb, preferably 50 to 2500 ppb, more preferably 100 to 1500 ppb, even more preferably 200 to 800 ppb of denatonium benzoate in the shoot tissue of the seedling; or a seedling containing an average concentration of 100 to 200 ppb, preferably 100 to 300 ppb, even more preferably 200 to 800 ppb, in the leaf tissue of the seedling. A seedling containing denatonium benzoate at a concentration of 5 to 50,000 ppb, preferably 5 to 25,000 ppb, more preferably 5 to 10,000 ppb, even more preferably 5 to 5,000 ppb or 5 to 2500 ppb or 5 to 1000 ppb; or a plant at development stage V6-V8 containing an average concentration of 0.5 to 200 ppb, preferably 0.5 to 100 ppb, more preferably 1 to 50 ppb, even more preferably 5 to 20 ppb of denatonium benzoate in the leaf tissue of the plant. Alternatively, the plant may be, for example, a seedling containing an average concentration of at least 1 ppm, at least 2 ppm, at least 3 ppm, at least 5 ppm, at least 10 ppm, at least 20 ppm, at least 30 ppm, at least 50 ppm, at least 100 ppm or at least 200 ppm of denatonium benzoate throughout the seedling, or an average concentration of at least 50 ppb, at least 75 ppb, at least 100 ppb, at least 150 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 750 ppb, at least 1000 ppb, at least 200 ppb, at least 300 ppb, at least 400 ppb, at least 500 ppb, at least 600 ppb, at least 750 ppb, at least 800 ppb, at least 900 ppb, at least 1000 ppb, at least 1200 ppb, at least 1500 ppb, at least 1600 ppb, at least 1700 ppb, at least 1800 ppb, at least 1900 ppb, at least 20 ... or seedlings having an average concentration of at least 5 ppb, at least 10 ppb, at least 20 ppb, at least 30 ppb, at least 50 ppb, at least 100 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 1000 ppb, at least 2000 ppb, at least 3000 ppb, at least 5000 ppb, at least 10000 ppb, at least 20000 ppb, at least 30000 ppb, or at least 50000 ppb of denatonium benzoate in leaf tissue of the seedlings.

[0053] The plant may be, for example, a corn seedling containing an average concentration of 1 to 200 ppm or 1 to 100 ppm, preferably 1 to 50 ppm, more preferably 5 to 30 ppm, even more preferably 10 to 22 ppm of denatonium benzoate in the entire seedling; or a corn seedling containing an average concentration of 50 to 4000 ppb, preferably 50 to 2500 ppb, more preferably 100 to 1500 ppb, even more preferably 200 to 800 ppb of denatonium benzoate in the shoot tissue of the seedling. ; or corn seedlings having an average concentration of 1 to 2000 ppb, preferably 1 to 1500 ppb, more preferably 1 to 1000 ppb, even more preferably 1 to 500 ppb of denatonium benzoate in the leaf tissue of the seedlings; or corn plants at developmental stages V6-V8, having an average concentration of 0.5 to 200 ppb, preferably 0.5 to 100 ppb, more preferably 1 to 50 ppb, even more preferably 5 to 20 ppb of denatonium benzoate in the leaf tissue of the plant. Alternatively, the plant can be, for example, a corn seedling containing an average concentration of at least 1 ppm, at least 2 ppm, at least 3 ppm, at least 5 ppm, at least 10 ppm, at least 20 ppm, at least 30 ppm, at least 50 ppm, at least 100 ppm, at least 150 ppm or at least 200 ppm of denatonium benzoate throughout the seedling, or an average concentration of at least 50 ppb, at least 75 ppb, at least 100 ppb, at least 150 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb in the shoot tissue of the seedling. ppb, at least 750 ppb, at least 1000 ppb, at least 2000 ppb or at least 4000 ppb of denatonium benzoate, or corn seedlings containing an average concentration of at least 1 ppb, at least 2 ppb, at least 3 ppb, at least 5 ppb, at least 10 ppb, at least 20 ppb, at least 30 ppb, at least 50 ppb, at least 100 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 1000 ppb or at least 2000 ppb of denatonium benzoate in leaf tissue of the seedlings.

[0054] Plants can be, for example, rape seedlings containing an average concentration of 10 to 20000 ppb, preferably 10 to 15000 ppb or 10 to 10000 ppb, more preferably 20 to 7500 ppb, even more preferably 20 to 5000 ppb of denatonium benzoate in the leaf tissue of the seedlings. Alternatively, plants can be, for example, rape seedlings containing an average concentration of at least 10 ppb, at least 20 ppb, at least 30 ppb, at least 50 ppb, at least 100 ppb, at least 200 ppb, at least 300 ppb, at least 500 ppb, at least 1000 ppb, at least 2000 ppb, at least 3000 ppb, at least 5000 ppb, at least 10000 ppb, at least 15000 ppb or at least 20000 ppb of denatonium benzoate in the leaf tissue of the seedlings.

[0055] Plants can be, for example, rye seedlings containing an average concentration of 1 to 200 ppm or 1 to 100 ppm, preferably 1 to 50 ppm, more preferably 5 to 30 ppm, even more preferably 10 to 22 ppm of denatonium benzoate in the leaf tissue and / or root tissue of the seedlings. Alternatively, plants can be, for example, rye seedlings containing an average concentration of at least 1 ppm, at least 2 ppm, at least 3 ppm, at least 5 ppm, at least 10 ppm, at least 20 ppm, at least 30 ppm, at least 50 ppm, at least 75 ppm, at least 100 ppm, at least 150 ppm or at least 200 ppm of denatonium benzoate in the leaf tissue and / or root tissue of the seedlings.

[0056] The plant may for example be any seedling as further described above.

[0057] Under bird infestation conditions, the seeds and plants according to the invention are protected from bird damage due to the presence of denatonium benzoate on and / or in the seeds or plants, which results in the birds being repelled away from the seeds or plants. In addition, the seeds and plants according to the invention are protected from damage caused by insects, particularly nematodes, due to the presence of denatonium benzoate on and / or in the seeds or plants, which results in the insects being repelled away from the seeds or plants. In agricultural areas, these plants will grow better than plants grown from untreated seeds.

[0058] Therefore, a further aspect of the invention is a method for controlling damage to seeds or plants in an agricultural area by birds and / or insects, the method comprising the steps of:

[0059] a) sowing seeds according to the invention or seeds obtained or obtainable by the method according to the invention in an agricultural area, and

[0060] a) growing plants from seeds,

[0061] wherein one or both of steps a) and b) are carried out under conditions of bird and / or insect infestation;

[0062] wherein the plant stand count in the agricultural area is higher than the plant stand count in a similar agricultural area having untreated seeds or plants grown from untreated seeds under the same conditions.

[0063] The seeds sown in step (a) do not need to be treated with denatonium benzoate immediately before sowing, but seeds which have been treated at an earlier stage and then stored may be used.

[0064] The conditions for growing the plants in step b) correspond to the conditions usually applied in cultivation. The fact that the seeds have been treated with denatonium benzoate does not impose any special requirements on the growing conditions.

[0065] "Stand counts" analyze the number of plants or crops on a field or a specific area of ​​a field. In a preferred embodiment, the plant stand counts of the agricultural area under conditions of bird and / or insect infestation are at least 5% or at least 10%, preferably 20% and more preferably at least 30% higher than the plant stand counts of a similar agricultural area obtained under the same conditions with untreated seeds or plants grown from untreated seeds.

[0066] The term "similar agricultural areas having untreated seeds or plants grown from untreated seeds" refers to areas where plants are grown under the same conditions (e.g., with respect to soil conditions, light, water, nutrients, etc.) and under the same conditions of bird and / or insect infestation, with the only difference being that the seeds or plants grown from untreated seeds have not been treated to protect them from bird and / or insect infestation.

[0067] The present invention will be further described by the following figures and examples.

[0068] Attached photos

[0069] Figure 1A Shown are plant stand counts at various test locations for corn seeds treated with denatonium benzoate or combinations of denatonium benzoate with various common fungicides compared to corn seeds treated with common bird repellents and controls

[0070] Figure 1B Plant stand counts summed over all locations are shown.

[0071] Figure 2 A shows a series of plant pots in which four corn seeds have been sown in each pot.

[0072] Figure 2 B shows the placement of a nematode of the genus Tapeworm in the central cavity of a plant pot.

[0073] Figure 3 is a bar graph showing plant biomass of maize plantlets developed from differently treated seeds (Variants 1-10) in the greenhouse.

[0074] 1: Negative control (no seed treatment)

[0075] 2: Adhesive alone (control)

[0076] 3.4: Bitrex

[0077] 5: RedigoM

[0078] 6-8: Bitrex and RedigoM

[0079] 9: RedigoM and Force 20CS (positive control)

[0080] 10: RedigoM and Poncho (positive control)

[0081] Figure 4 : Bar graph showing the assessment of seed damage caused by nematodes on maize plantlets developed from differently treated seeds (variants 1-10) in a greenhouse (A) and a phytotron (B).

[0082] Figure 5 Analysis of variance of plant stand counts at three different locations (AC) is shown. Denatonium Benzyl (DNB) alone or in combination with a micronutrient solution mixed with a sticker (DNB+HA450) was applied to the seeds. Three chemical references were included in the trial: Korit, Sphere and Schwefal. Control: Untreated seeds Example 1. Evaluation of efficacy as a bird repellent.

[0083] In order to evaluate the efficacy of different substances as bird repellents, 25 locations throughout Europe were identified to have potential high bird damage pressure. 17 different substances (DSM RD9103 BRP, pepper extract, black cumin oil, yucca extract, labial mint extract, aluminum ammonium sulfate, hop extract, fat-encapsulated plant extract, alginate-encapsulated plant extract, tefluthrin, Thiram, Ziram, Methiocarb, yellow substance, benzyl denatonium, SatecWFO20, Redigo M, Maxim XL) have been selected to evaluate the difference in crow feeding behavior. Three chemical references are included in the test: Mesurol liquid (active substance Methiocarb), Korit (active substance Ziram) and TMTD (active substance Thiram). Mesurol liquid (Methiocarb) is a standard bird repellent in the past, and the expiration date in Europe is April 3, 2020. TMTD (Thiram) can be used until January 30, 2020. Korit (Ziram) expires in 2021.

[0084] Denatonium benzoate was tested at 2 different concentrations: 0.4 g / 50.000 seeds in combination with the standard fungicides Redigo M and Maxim XL, and 0.8 g / 50.000 seeds alone. In all combinations and variations, a film coating (adhesive) has been used to provide a uniform distribution of the different substances to the seeds.

[0085] In one embodiment, denatonium benzoate is used in the form of a solid powder in a crystalline pure form (CAS: 3734-33-6; ai=3500 ppm). In another embodiment, the denatonium benzoate powder is preferably mixed with a sticking agent before application, and then preferably applied at a concentration of 13.14 g per liter to 26.28 g per liter.

[0086] Table 1. Exemplary compound mixtures in seed treatments:

[0087]

[0088] Specifically, the adhesive is selected from Quality or Active, particularly preferably the adhesive is selected from Sepiret 9290red or Quality 316 https: / / www.seedgrowth.bayer.com / ~ / media / d3ed77a4560740d8a568fa06f50129dc.ashx. Other possible adhesives can be found at https: / / agriculture.basf.com / en / Crop-Protection / Seed-treatment.html.

[0089] The treatment process can be based on the rotor / stator principle. The seeds are accelerated by the rotating mixing cone and moved to the top of the static cylinder wall. Special bypass blades on the top of the wall guide the rising seeds back to the middle. Thus, a fine seed curtain is established around the chemical liquid spray disc. The spray disc produces a fine mist of chemical liquid, which covers the inner seed curtain as well as the rising seeds at the cylinder wall.

[0090] For the field trials, three different types of experimental arrangements were arranged: completely randomized microplot trials at 4 locations, non-randomized 8-row plots with three replications of untreated controls within the plots at two locations; non-randomized 2-replication 1-row plots with three untreated controls at four locations; and non-randomized 1-row plots with five replications of untreated controls at 16 locations.

[0091] Three weeks after emergence, nine locations affected by bird damage were identified. Plant stand counts were assessed and summarized. Figure 1A The data for each test location are shown. Figure 1B Plant stand counts summed over all locations are shown.

[0092] Surprisingly, it has been found that the use of denatonium methyl as a Corvidae bird repellent for corn has shown excellent results. The efficiency is similar to or even better than the best bird repellents such as Thiram, Korit and Mesurol. In addition, it has been observed that denatonium methyl has a systemic effect on corn plants. This not only protects the seeds from the infringement of birds, but also protects the seedlings that grow. Since denatonium methyl is diluted during growth and maturity, the crops (e.g. corn cobs) harvested do not show the amount of denatonium methyl that affects the use of corn after harvest (e.g., for animal feeding or silage purposes).

[0093] The bird repellent effect of denatonium benzoate was further evaluated in additional field trials at three different locations in Germany. Denatonium benzoate (DNB) alone or in combination with a micronutrient solution mixed with an adhesive (DNB+HA450) was applied to the seeds. The application concentration of denatonium benzoate was 1 g / 50.000 seeds. Three chemical references were included in the test: Korit (active substance: Ziram), Curb Crop Garden powder with Sphere's formulation (active substance: aluminum ammonium sulfate) and Schwefal (active substance: organic sulfur). As a negative control, untreated seeds were used. All three locations showed moderate to high levels of bird damage.

[0094] Plant stand counts were evaluated and summarized. Figure 2 AC shows the analysis of variance for each test location. The results of the earlier field trials were confirmed. It was found that the use of denatonium benzoate as a corvid bird repellent for corn again showed excellent results. The efficiency was similar to the best bird repellents such as Korit and Sphere. In the field trials, the use of Schwefal on corn seeds did not show any significant bird repellent effect.

[0095] Example 2. Phytotoxicity Evaluation

[0096] To evaluate the phytotoxicity of different corn seed treatments, a safety test was performed. After five days at 25°C, the seeds were treated with different concentrations of denatonium benzoate. [g / unit, where 1 unit is 50.000 seeds] The germination rate of corn seeds treated with 100% chlorinated benzyl alcohol was counted. The cold test germination rate (CT) was counted after 7 days at 10°C and 5 days at 25°C. The treated and untreated seeds were compared. In addition, the germination rate of corn seeds treated with denatonium benzoate in combination with further active agents, in particular fungicides, was examined under different environmental conditions. The results are shown in the following Table 2, where

[0097] KF = germination percentage counted after 5 days at 25°C, and

[0098] CT = chill test germination percentage counted after 7 days at 10°C and 5 days at 25°C.

[0099] Table 2

[0100]

[0101]

[0102]

[0103] Example 3: Determination of systemic activity of denatonium benzoate

[0104] To determine whether denatonium benzoate could be systemic (absorbed by the plant), liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) was performed, which is a powerful analytical technique that combines the separation power of liquid chromatography with the high sensitivity and selective mass analysis capabilities of triple quadrupole mass spectrometry. Therefore, the inventors used 1) seedlings after field emergence to determine the content of denatonium benzoate in whole plants and in shoots alone (Table 3), and 2) middle-aged plants (V6-V8), where leaves were studied (Table 4).

[0105] Table 3. Denatonium benzoate determination results for seedlings after field emergence

[0106]

[0107] Table 4. Results of determination of denatonium benzoate in middle-aged plants

[0108]

[0109] Denatonium benzoate is absorbed by plants and shows differences in concentrations between different plant parts / organs and developmental stages:

[0110] In the early stages, denatonium benzoate was selectively detectable. The concentration in the whole plant was in the lower ppm range (about 10-20 ppm), and the concentration in the bud was in the higher ppb range (about 200-800 ppb). The concentration varied greatly (Table 3). In middle-aged corn plants, the concentration in the leaves was in the lower ppb range (about 5-20 ppb). The variation in concentration was different between types (between) and plants (within), and the concentration of DNB decreased significantly from the early stage (200-800 ppb in the bud) to the mid-stage (5-20 ppb in the leaf).

[0111] In addition to the above experiments, the concentration of Bitrex in the first leaves of seedlings was measured, in which different doses of Bitrex were applied to the seeds. The higher the application rate, the higher the Bitrex concentration in the tested leaf tissue.

[0112] Table 4a. Results of determination of denatonium benzoate in seedling leaves

[0113]

[0114] Further experiments on systemic uptake of Bitrex were also conducted on rapeseed (Brassica napus). Brassica napus seeds were treated with Bitrex at concentrations ranging from 0.15 g / unit to 75 g / unit. Seeds were sown and leaf samples were collected 6 days after emergence and 22 days after emergence. The amount of Bitrex in leaf tissue was determined by liquid chromatography combined with tandem mass spectrometry (LC-MS-MS) as described above, with 4 replicates per Bitrex concentration. Leaf samples were prepared as follows: leaf material was harvested, cut into small pieces and placed in 3-5 ml of ethanol. After vortexing for 45 minutes, centrifuged and 1 ml aliquots were taken for further analysis.

[0115] Table 4b: Denatonium benzoate determination results for seedlings. One unit of rapeseed is 1.5 million viable seeds, which is typically about 7 kg of seeds per unit.

[0116]

[0117]

[0118] The results (Table 4b) show that Bitrex has been absorbed into the tissues of the seedlings. The amount of Bitrex in the leaf tissue depends on the amount of Bitrix applied to the seeds. Interestingly, the uptake of Bitrex is quite similar after 6 days at concentrations of 0.15-7.5 g / unit. At concentrations of 15 g / unit and 75 g / unit, significantly higher amounts of Bitrex were observed. After 22 days, Bitrex was still detectable for all applications, but in lower amounts. This indicates that Bitrex is further distributed in the plantlets and is also degraded. However, especially in the early seedling stage, the amount of Bitrex in the plants is quite high and provides the desired repellent effect against birds and insects and reduces the corresponding damage. At the same time, due to further distribution and degradation, adult plants contain almost no Bitrex.

[0119] Additional experiments conducted in hybrid rye also showed excellent systemic uptake and distribution of Bitrex. Seeds of hybrid rye varieties were treated with KintoPlus 150 ml / 100 kg seed and Bitrex 75 g / 100 kg seed in Application A and KintoPlus 150 ml / 100 kg seed and Bitrex 150 g / 100 kg seed in Application B. Seeds were sown in longitudinal plots of 15 qm in four replicates. Seedlings (40 plants per plot) were harvested 3 weeks after sowing and leaf material and root tissue were analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) as described above.

[0120] Table 4c: Denatonium benzoate determination results for seedlings. One unit of rapeseed is 1.5 million viable seeds, which is typically about 7 kg of seeds per unit.

[0121]

[0122] The results in Table 4c show that rye plants absorb Bitrex in tissues such as roots and leaves. The applied dose of Bitrex on the seeds determines the subsequent concentration in the tissues. This clearly indicates that in rye, Bitrex is also distributed systemically. In this experiment, concentrations of Bitrex in leaves between 3,000 ng / g and 30,000 ng / g and in roots between 4,500 ng / g and 25,000 ng / g were measured. As expected, in the grains, the concentrations were even higher, up to 460,000 ng / g. Example 4: Evaluation of the efficacy of denatonium benzoate as a nematode repellent in greenhouses and climatic chambers The following experimental protocol was arranged as shown in Table 5. Denatonium benzoate (Bitrex) was tested in concentrations of 0.5 g / unit (where 1 unit = 50.000 seeds), 1 g / unit and 5 g / unit, without any other plant protection product (variants 3 and 4) or with RedigoM (active ingredients 100 g / l prothioconazole (PRO) and 20 g / l metalaxyl (MTL); (variants 6-8). The adhesive was always Sepiret 9290 red, whereby 30 ml / unit was applied. Variant 1 was a negative control in which the seeds were not treated, variant 2 was a further control treated only with the adhesive. Variant 5 was RedigoM and an adhesive without Bitrex. Variants 9 and 10 were used as positive controls. In addition to RedigoM and the adhesive, Force 20 CS (active ingredient: tefluthrin (TEF); or with 600 (active ingredient: clothianidin (CLO) for seed treatment. Force 20 CS and 600 are standard treatments against nematode infestations. All tests were conducted on the maize variety Amavit from KWS SAATSE & Co. KGaA.

[0123] Table 5: Setup for nematode repellent testing. Variants 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 were tested in the greenhouse and in the climatic chamber, and variant 4b was tested only in the climatic chamber.

[0124]

[0125] The test was conducted in a pot assay. Corn seeds were sown in each pot (see Figure 2A). In each pot, four nematodes of the genus Ceratitis were placed in a central cavity 5 cm deep (see Figure 2 B) Each treatment variant 1-10 was replicated seven times. Seeds in pots were germinated and seedlings were grown under controlled conditions in a greenhouse.

[0126] Eight days after sowing, plant biomass ( Figure 3 ; Table 6) and seed damage ( Figure 4 A; Table 6). The treatment with Bitrex had a positive effect on the development of biomass. Plantlets not treated with plant protection products (variants 1 and 2) suffered from infection and showed slower early plant development. Treatment with the adhesive Seperit 9290 red alone (variant 2) had no effect on plant development. Plantlets treated with Bitrex showed a similar or even greater biomass than control plants without nematode infestation ("Control (without WW)") and similar to variants 9 and 10 treated with the standard plant protection products Force 20CS and Poncho.

[0127] If Bitrex is applied, the evaluated seed damage and seedling damage (preferably root damage) are significantly reduced. Untreated seeds (variant 1) showed that an average of 3 seeds out of four sown seeds per pot were damaged (average damage: 75%). As positive controls, Force 20CS and Poncho showed no seed damage at all (average damage: 0%). The application of Bitrex, especially in concentrations above 1 g / unit, significantly increased the tolerance to nematodes (see variants 4, 7 and 8). In variant 8, seed damage could be reduced to an average of 21.5% (corresponding to an average of 0.86 damaged seeds out of four seeds per pot).

[0128] The test was repeated in a climatic chamber in order to ensure high humidity and moderate temperature, which provide optimal environmental conditions for the nematodes. Thus, a high level of insect pressure on the sown maize should be simulated. In each pot, four nematodes of the genus Ceratoides were placed in a central cavity 5 cm deep (see Figure 2 B). Each treatment variant 1-10 was replicated eight times (8 pots, each with 4 seeds). The seeds in the pots were germinated and the seedlings were grown under phytotron conditions: 18°C ​​day / 12°C night.

[0129] Eighteen days after sowing, the biomass of plants (Table 6) and seed damage ( Figure 4 Even under such enhanced biotic stress conditions, the positive effects of Bitrex on biomass development as well as seed and seedling damage were confirmed.

[0130] Table 6: Results of tests in a greenhouse (GH) and a climate chamber (CC)

[0131]

[0132] Example 5: Evaluation of the efficacy of denatonium benzoate as a nematode repellent in field trials

[0133] In order to determine whether a positive effect of Bitrex on biomass development and on seed damage and seedling damage could also be expected in the field, a similar experimental arrangement as described for the greenhouse and climatic chamber described above was used, except that the application rate of Bitrix was different. Seeds were treated with 5 g Bitrex per 50,000 seeds, 10 g Bitrex per 50,000 seeds and 20 g Bitrex per 50,000 seeds. At one location in Italy, 4 rows (21 m 2 ) in a plot where the soil showed high exposure to the genus Ceratitis nematodes. Evaluations were performed at emergence (E), at the V3 stage (= 3 leaves unfolded) and at the V5 stage (= 5 leaves unfolded). The following characteristics were phenotyped: phytotoxicity (PHYGEN) at the E, V3 and V5 stages in % / plot; vigor (VIGOR) at stages E, V3 and V5 on a scale of 1-10 (10 being the best); insect damage (DAMINS) at stage V5 in number of plants / row (left DAMINS column in Table 7) and in % / row (right DAMINS column in Table 7). Force 20CS was again used as a positive control (60 ml / 50,000 seeds) and untreated seeds (Control) were used as negative controls.

[0134] Table 7: Test results of field trials.

[0135]

[0136] Field trials have shown that under natural conditions, treatment of seeds with Bitrex also has a repellent effect on nematodes. In this experiment, the application of 5-20 g of Bitrex reduced the number of corn plants affected by nematode damage by about 35%-45%. Even when Bitrex was applied at high concentrations, no negative effects on plant vitality related to phytotoxicity and plant vigor were observed. Thus, the trials have shown that under field conditions, Bitrex can also be used as an insect repellent, which presents a new ecologically safe alternative to other chemicals that may be harmful to the environment.

Claims

1. Use of denatonium benzoate as the sole active ingredient for protecting seeds, plants, seedlings from damage caused by birds of the genus Corvidae or nematodes by repelling them, the denatonium benzoate optionally in combination with one or more adhesion additives, the seeds, plants, seedlings being seeds, plants, seedlings of corn, rapeseed or rye.

2. The use of claim 1, wherein the adhesion additive comprises a sticky agent that improves the adhesion of denatonium benzoate to plant seeds.

3. The use of claim 1, wherein the adhesion additive comprises a natural resin-based adhesive, a lignin sulfonate-based adhesive, a polymer-based adhesive and / or a water-based adhesive.

4. The method of claim 3, wherein the water-based adhesive is Quality or Active.

5. A method for protecting seeds from damage caused by corvids or nematodes by repelling corvids or nematodes, the method comprising treating the seeds with denatonium benzoate as the sole active ingredient, optionally in combination with one or more adhesion additives, the seeds being seeds of corn, rapeseed or rye.

6. The method of claim 5, wherein the adhesion additive comprises a sticking agent that enhances the adhesion of denatonium benzoate to the plant seeds.

7. The method of claim 5, wherein the adhesion additive comprises a natural resin-based adhesive, a lignin sulfonate-based adhesive, a polymer-based adhesive, and / or a water-based adhesive.

8. The method of claim 7, wherein the water-based adhesive is Quality or Active.

9. The method according to any one of claims 5 to 8, wherein the amount of denatonium benzoate in corn is 0.1 to 70 g / 50,000 seeds, 0.1 to 50 g / 50,000 seeds, 0.1 to 20 g / 50,000 seeds, 0.1 to 10 g / 50,000 seeds or 0.1 to 5 g / 50,000 seeds; or wherein the amount of denatonium benzoate in rapeseed is 2 to 200 g / 1.5 million rapeseed seeds, 2 to 100 g / 1.5 million rapeseed seeds, 2 to 50 g / 1.5 million rapeseed seeds, 2 to 20 g / 1.5 million rapeseed seeds or 2 to 10 g / 1.5 million rapeseed seeds; or wherein the amount of denatonium benzoate in rye is 10 to 1500 g / 100 kg rye seeds, 10 to 1000 g / 100 kg rye seeds, 10 to 750 g / 100 kg rye seeds, 10 to 500 g / 100 kg rye seeds or 250 g / 100 kg rye seeds.

10. The method according to claim 9, wherein the amount of denatonium benzoate used in corn is 0.2 to 5 g / 50,000 seeds.

11. The method according to claim 9, wherein the amount of denatonium benzoate used in rapeseed is 2 to 25 g / 1.5 million rapeseed seeds.

12. The method according to claim 9, wherein the amount of denatonium benzoate in rye is 10 to 75 g / 100 kg rye seeds or 10 to 150 g / 100 kg rye seeds.

13. The method according to any one of claims 5 to 8, wherein the denatonium benzoate is used in the form of a solid powder in a crystalline pure form or in the form of a liquid composition further comprising a binder.

14. The method of claim 13, wherein the liquid composition comprises denatonium benzoate at a concentration of 1-800 g / l, 1-500 g / l, 1-250 g / l, 1-100 g / L, 10-50 g / L, or 13-27 g / L.

15. The method of claim 13, wherein the liquid composition comprises denatonium benzoate at a concentration of 10-50 g / L.

16. The method of claim 13, wherein the liquid composition comprises denatonium benzoate at a concentration of 13-27 g / L.

17. A method of providing a plant protected from damage caused by corvids or nematodes by repelling corvids or nematodes, the method comprising a) providing a treated seed obtained according to the method of any one of claims 5 to 16, and b) growing plants from the treated seeds of a).

18. A method for controlling damage to seeds or plants in an agricultural area caused by corvids or nematodes by repelling corvids or nematodes, the method comprising the steps of: a) sowing the treated seeds obtained according to any one of claims 5 to 16 in an agricultural area, and b) growing plants from said treated seeds, wherein one or both of steps a) and b) are carried out under conditions of infestation by corvids or nematodes; wherein the plant stand count of the agricultural area is higher than the plant stand count of a similar agricultural area having untreated seeds or plants grown from untreated seeds under the same conditions.

19. The method of claim 18, wherein the plant stand count of the agricultural area is at least 5% or at least 10% greater than the plant stand count of a similar agricultural area having untreated seeds or plants grown from untreated seeds.

20. The method of claim 18 or 19, wherein the plant stand count of the agricultural area is at least 20% greater than the plant stand count of a similar agricultural area having untreated seeds or plants grown from untreated seeds.

21. The method of claim 18 or 19, wherein the plant stand count of the agricultural area is at least 30% greater than the plant stand count of a similar agricultural area having untreated seeds or plants grown from untreated seeds.

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