Tobacco alkaloid insecticide in plants

Abstract

Use of a formulation for preventing or reducing the infestation of Hemiptera insects, wherein the formulation comprises a compound of formula I.

Description

Technical Field

[0001] In its broadest aspects, the present disclosure relates to the use of formulations comprising a compound of formula I for preventing Hemiptera insects. Methods for preventing Hemiptera insects are also provided herein.

Background Art

[0002] Agriculture is an important activity for humans, providing basic needs such as food, clothing, and shelter. Agricultural productivity can be adversely affected by many factors, including insect infestations. Insect infestations can impede crop and plant growth and damage cultivated and natural plants. The interference and damage can cause the crops to wither and prevent the plants from maturing. Insect infestations are estimated to destroy at least 30-40 percent (%) of world agricultural production and pose a particular threat to parts of the world economy that depend on agriculture.

[0003] Insects are responsible for two main types of damage to cultivated crops. First, insect feeding causes direct damage to plants by eating leaves or entering the plant's stem, fruit, or roots. Second, there can be indirect damage when the insects themselves cause little or no harm to the plants but transmit bacterial, viral, or fungal infections to the crops.

[0004] Whiteflies are Hemiptera insects belonging to the Aleyrodidae family. They are considered major pests of many crops because they cause significant damage and reduction in yields. They obtain nutrients by sucking sap from host plants. They are polyphagous, meaning they feed on many different plants and pose a danger to most crops.

[0005] Two common species of whiteflies are the tobacco whitefly (Bemisia tabaci) and the greenhouse whitefly (Trialeurodes vaporariorum). Bemisia tabaci is currently recognized as a complex of approximately 40 cryptic species distributed worldwide and can feed on over 900 plant species, including a wide range of vegetables (such as tomatoes, eggplants, sweet potatoes, okra, broccoli, cabbages, etc.), as well as cotton, tobacco, and ornamental plants such as poinsettias, garden roses, lantanas, or lilies.

[0006] The entire life cycle of whiteflies lasts from 15 to 40 days, depending on the host plant and environmental conditions. For example, the number of eggs laid, as well as the survival rates of the immature stages and adults, are greatly affected by temperature, and the optimal reproductive adaptation is at approximately 28 - 30 degrees Celsius (°C) (Curnutte et al., Annals of the Entomological Society of America, Vol. 107, No. 5).

[0007] Whiteflies lay eggs on both the upper and lower sides of leaves. From the hatched eggs, the first larval instar (crawler) emerges and moves along the leaf until it finds a suitable place to start feeding on the phloem sap, which is rich in sugar. The larvae mature into the second, third, and fourth larval instars, during which they remain in the same place and continue to feed on the plant. The fourth instar stage is also called the pupal or red larval stage from which the adults emerge (Sani et al., Insects, 2020, 11, 619).

[0008] Direct damage is caused to the plant by the feeding of whiteflies. The sucking of sap leaves discolored spots on the leaves. Furthermore, during the sucking of sap, they release toxic substances into the phloem, which spreads throughout the plant. This leads to metabolic imbalance, causing overall weakening, whitening of the plant, as well as changes in flowers and fruits.

[0009] From the perspective of indirect damage, the honey excreted by the larvae causes fungi such as sooty mold to form on the leaves. This mold acts as a screen and reduces the photosynthetic ability of the plant.

[0010] Finally, the most serious damage caused by whiteflies to crops is virus transmission. The B. tabaci complex is known to vector over 200 plant viruses. These include TYLCV (Tomato Yellow Leaf Curl Virus), TYMV (Tomato Yellow Mosaic Virus), or the viruses of the cotton leaf curl disease complex.

[0011] One of the main objectives in controlling whiteflies is to avoid the crop being infected by viruses. The use of protective barriers such as nets and covers is a good option for preventing entry, but they are only suitable for smaller and less mature crops.

[0012] The use of control agents is the most effective way to prevent entry throughout the life of the crop.

[0013] Most insecticides are synthetic pesticides. Due to the long-term use of such agents, many pests have developed resistance. Resistance is defined as a change in the susceptibility of a pest population to an insecticide, resulting in the failure of the correct application of the insecticide. The intensive use of insecticides over the past decade to control whiteflies has led to the emergence of populations resistant to various classes of insecticides such as carbamates, organophosphates, and pyrethroids.

[0014] In addition to the difficulty of controlling these pests with current insecticides, some insecticides are highly toxic and there is increasing evidence that they take years to biodegrade. Therefore, it is desirable to develop new, naturally derived insecticides with a mode of action different from those known. Furthermore, naturally derived insecticides have a lower impact on the environment and are generally safer and less hazardous for those who use them.

[0015] Among naturally occurring insecticides, tobacco (Nicotiana genus spp.) plants have been used for centuries in the form of dried leaves, powders, extracts, or fumigants. However, due to concerns about the safety of nicotine, the main alkaloid in tobacco, and the discovery of more specific and potent synthetic insecticides, interest in tobacco has declined. As a result, tobacco-derived products or nicotine-based products are not currently commercially available.

[0016] Therefore, there is a clear need to identify more plant-derived compounds that are as effective as synthetic insecticides but are more environmentally friendly, safer, and have novel modes of action for inclusion in agricultural crop pest management programs. It is desirable to provide new compounds and formulations that act in a different way than killing insects and are particularly active as insecticides. SUMMARY OF THE INVENTION

[0017] The present disclosure relates to the use of a formulation for preventing or reducing the infestation of Hemiptera insects. The formulation may contain a compound of Formula I. The compound of Formula I may be as follows,

Chemical formula

Chemical formula

[0018] According to one aspect of the present invention, there is provided the use of a formulation for preventing Hemiptera insects. According to another aspect of the present invention, there is provided the use of a formulation for reducing the infestation of Hemiptera insects. The formulation contains a compound of Formula I. The compound of Formula I is as follows,

Chemical formula

Chemical formula

[0019] As used herein, the terms "insect repellent", "insect repellent effect", or "insect repellent efficacy" mean the effect of preventing insects from staying away from and / or landing on, walking on, contacting, or laying eggs on a surface or substance.

[0020] As used herein, the term "infestation" means a state in which an abnormally large number of insects or other species are present on a surface or substance.

[0021] As used herein, the term "insect" means any member of the largest class of the phylum Arthropoda.

[0022] As used herein, the term "hemipteran insect" means an insect belonging to a taxonomic rank generally called true bugs, which share a common arrangement of sucking mouthparts. Thus, as used herein, the term "hemipteran insect" is intended to define an insect that feeds on plants and extracts sap from plants using sucking and piercing mouthparts.

[0023] Here, the present invention will be further described with reference to the drawings.

Brief Description of the Drawings

[0024]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

[0025] As described above, the present invention relates to the use of a formulation for preventing or reducing the invasion of Hemiptera insects.

[0026] The use of a formulation containing a compound of formula I has shown an improved insect repellent effect against Hemiptera insects, particularly Bemisia tabaci.

[0027] In particular, the use of a formulation containing anatabine has shown an improved insect repellent effect against Hemiptera insects, particularly Bemisia tabaci.

[0028] The insecticidal effect of the formulation according to the present invention has been shown to be stronger and more persistent than commonly used chemical agents such as DEET (N,N-diethyl-meta-toluamide). This has the advantage of reducing the amount and frequency with which the formulation has to be applied to crops or plants. Furthermore, the insecticidal effect has been found to be more effective than methyl benzoate, a natural plant-derived compound known in the art to be effective against Bemisia tabaci (Mostafiz et al., PLoS ONE 13, 2018). Surprisingly, it has been found that the insecticidal effect of the formulation according to the present invention can be maintained at 100 percent over a long period without damaging the crop or plant. The absence of damage to the crop or plant is evidence that the effectiveness of the formulation according to the present invention is directly due to driving away the hemipteran insects from their food source rather than destroying it. This insecticidal effect has the additional advantage of preventing oviposition on the leaves. The combination of the effect of preventing access to food and the effect of preventing oviposition results in an overall insecticidal effect. This overall insecticidal effect has been shown to last surprisingly long, ultimately resulting in the disruption of the life cycle of Bemisia tabaci and consequently a reduction in the Bemisia tabaci population in the area where the formulation is applied.

[0029] Some tobacco plant compounds such as nicotine are known to be able to provide a defensive barrier against insect invasion. Nicotine is one of the most abundant compounds in tobacco plants and has been reported to be highly toxic to most insects that feed on the plants. However, Bemisia tabaci shows significant tolerance to these and other tobacco plant compounds to the extent that it can feed on tobacco plants and consume these compounds at levels 40 times higher compared to other insects. Surprisingly, it has been found that tobacco plants, particularly natural compounds derived from anatabine, provide an insecticidal effect against hemipteran insects, particularly Bemisia tabaci, despite tobacco plants being a common food source for hemipteran insects. Advantageously, the use of anatabine and formulations containing anatabine provides a naturally derived insecticide that is very effective and efficient while being more environmentally friendly and safer to use.

[0030] In some embodiments of the present invention, there is provided the use of a formulation according to the present invention, wherein the Hemiptera insect may be an insect that feeds on sap. The insect that feeds on sap feeds on the leaves of a tree by ingesting the fluid component. The insect that feeds on sap has a straw or sucking mouthpart, which forms a mouth tip used to penetrate the tissue and suck from the phloem tissue embedded in the leaf. The nutritionally rich phloem tissue is a unique source of nutrition for insects that feed on sap, which have evolved mechanisms to gain access to it. The use of the formulation according to the present invention prevents Hemiptera insects from accessing this source of nutrition. Advantageously, this protects the source of nutrition from damage by insects that feed on sap and causes the insects that feed on sap to search for food elsewhere.

[0031] In a further embodiment, there is provided the use according to the present invention, wherein the Hemiptera insect may be a whitefly.

[0032] In other embodiments of the present invention, the Hemiptera insect may be selected from the family Aleyrodidae consisting of Aleyrodes proletella, Aleurocanthus spiniferus, Aleurolobus barodensis, Aleurothrixus floccosus, Bemisia tabaci, Bemisia argentifolii, Dialeurodes citri, Parabemisia myricae, Trialeurodes vaporarium, or Siphoninus phillyreae.

[0033] In other embodiments of the present invention, the hemipteran insects are Aphis spp., Acyrthosiphon spp, Anuraphis cardui, Aulacorthum solani, Brachycaudus helichrysi, Brevicoryne brassicae, Ceratovacuna lanigera, Chaetosiphon fragaefolii, Chromaphis juglandicola, Cryptomyzus ribis, Dysaphis spp, Myzus spp., Eriosoma spp., Hyalopterus arundinis, Lipaphis erysimi, Macrosiphum spp., Melanaphis sacchari, Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Nasonovia ribisnigri, Pemphigus spp., Phloeomyzus passerinii, Phorodon humuli, Rhopalosiphum spp., Schizaphis graminum, Tinocallis caryaefoliae, Toxoptera spp., Acizzia spp., Acrida turrita, Acrogonia spp., Aeneolamia spp., Agonoscena spp.Allocaridara malayensis, Amrasca spp., Aonidiella spp., Aphanostigma piri, Arboridia apicalis, Arytainilla spp., Aspidiella spp., Aspidiotus spp., Atanus spp., Balclutha spp., Blastopsylla occidentalis, Boreioglycaspis melaleucae, Cacopsylla spp., Calligypona marginata, Carneocephala fulgida, Cercopidae, Ceroplastes spp., Chionaspis tegalensis, Chlorita onukii, Chrysomphalus ficus, Cicadulina mbila, Coccomytilus halli, Coccus spp., Comstockaspis perniciosa, Cryptoneossa spp., Ctenarytaina spp., Dalbulus spp., Diaphorina citri, Diaspis spp., Drosicha spp., Dysmicoccus spp., Empoasca spp., Epiacanthus stramineus, Erythroneura spp., Eucalyptolyma spp., Euphyllura spp., Euscelis bilobatus, Ferrisia spp., Fiorinia theae, Geococcus coffeae, Glycaspis spp., Heteropsylla cubana, Heteropsylla spinulosa, Homalodisca coagulata, Icerya spp., Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lepidosaphes spp., Macrosteles facifrons, Mahanarva spp., Metcalfiella spp., Nephotettix spp., Nilaparvata lugens, Oncometopia spp., Orthezia praelonga, Pachypsylla spp., Paratrioza spp., Parlatoria spp., Peregrinus maidis, Phenacoccus spp., Phylloxera spp., Pinnaspis aspidistrae, Planococcus spp., Prosopidopsylla flava, Protopulvinaria pyriformis, Pseudaonidia paeoniae, Pseudaulacaspis ssp., Pseudococcus spp., Psyllopsis spp., Psylla spp., Pyrilla spp.It may be selected from the family Aphididae consisting of Quadraspidiotus spp., Quesada gigas, Rastrococcus spp., Saissetia spp., Scaphoideus titanus, Selenaspidus articulatus, Sogata spp., Sogatella furcifera, Sogatodes spp., Stictocephala festina, Tenalaphara malayensis, Tetragonocephela spp., Tomaspis spp., Trioza spp., Typhlocyba spp., Unaspis spp., Viteus vitifoliae, or Zygina spp..

[0034] In certain other embodiments of the present invention, the hemipteran insect may be selected from the group consisting of, but not limited to, Anasa tristis, Boisea spp., Blissus spp., Cavelerius spp., Cletus punctiger, Dasynus piperis, Diconocoris hewetti, Dysdercus spp, Eurygaster spp., Leptocorisa spp., Leptoglossus phyllopus, Macropes excavatus, Miridae, Pentastomidae, Piesma spp., Pseudacysta persea, Riptortus spp., Rhopalus spp., Scaptocoris castanea, Scotinophora spp., Stephanitis spp., or Togo hemipterus, which are heteropteran insects.

[0035] In some embodiments, there is provided a use according to the present invention, wherein the use may be for preventing or reducing the invasion of hemipteran insects from agricultural crops or plants.

[0036] In some further embodiments, agricultural crops or plants can be vulnerable to the invasion of whiteflies, and the agricultural crops or plants can be tobacco plants, cereals, legumes, oil plants, cucurbitaceae plants, fiber plants, vegetables, fruits, or flowers, and the use according to the present invention is provided.

[0037] In certain other embodiments of the present invention, the agricultural crops or plants can be selected from the group consisting of wheat, barley, rye, oats, corn, rice, sorghum, triticale, pome fruits, stone fruits, soft fruits, apples, grapes, pears, plums, peaches, almonds, pistachios, cherries, berries, strawberries, raspberries, blackberries, peppers, chili peppers, legumes, beans, lentils, peas, soybeans, oil plants, rape, mustard, sunflowers, marrows, cucumbers, melons, pumpkins, squashes, cotton, flax, hemp, jute, citrus fruits, calamondins, citrus citrons, citrus hybrids, kirs, tangelos, tangors, grapefruits, kumquats, lemons, limes, mandarins, sour oranges, sweet oranges, pomelos, satsumas, spinach, lettuce, asparagus, cabbages, carrots, onions, tomatoes, potatoes, paprika, broccoli, cauliflower, sweet potatoes, coffee, flowers, poinsettias, garden roses, lantanas, lilies, shrubs, broad-leaved trees, evergreen trees, coniferous trees, or Chinese hackberries.

[0038] In some embodiments of the present invention, the agricultural crops or plants can be found at all stages of their life cycle.

[0039] In a further embodiment, there is provided a use according to the invention, in which agricultural crops or plants can be cultivated in a greenhouse. The warm and humid conditions and abundant food inside the greenhouse provide an excellent and stable environment for the development of insects. Hemipteran insects, particularly whiteflies, thrive in temperate regions but are known to be able to develop easily in greenhouses located in conditions where they usually cannot maintain their life cycle. Advantageously, it has been found that the formulations according to the invention can be used in greenhouses. In fact, synthetic insecticides used in closed spaces bring serious harmful consequences to workers during and after application, while the use of the formulations according to the invention provides a safer and more environmentally sustainable natural alternative for users.

[0040] As detailed above, there is provided a use of a formulation for preventing or reducing the infestation of Hemipteran insects, the formulation comprising a compound of formula I,

Chemical formula

Chemical formula

[0041] In some embodiments of the invention, in the compound of formula I, R can represent hydrogen.

[0042] In some other embodiments of the invention, in the compound of formula I,

Chemical formula

[0043] In a particular embodiment of the invention, the compound of formula I may be anatabine.

[0044] Anatabine can occur in the form of two enantiomeric forms, S-(−)-anatabine and R-(+)-anatabine. Thus, anatabine can be present in the formulations of the present invention as either S-(−)-anatabine, R-(+)-anatabine, a mixture of S-(−)-anatabine and R-(+)-anatabine, or a racemate of S-(−)-anatabine and R-(+)-anatabine. The amounts of R-(+)-anatabine and S-(−)-anatabine may be present in any enantiomeric ratio, i.e., any enantiomeric excess (ee) of (R)-(+)-anatabine or S-(−)-anatabine.

[0045] As used herein, the term "enantiomeric ratio" of R-(+)-anatabine and S-(−)-anatabine is intended to mean the weight ratio of R-(+)-anatabine to S-(−)-anatabine.

[0046] According to the use of the present invention, the formulation may contain an enantiomeric ratio of R-(+)-anatabine to S-(−)-anatabine of 10:90, 20:80, 30:70, 40:60, 60:40, 70:30, 80:20, or 90:10.

[0047] The enantiomeric ratio of R-(+)-anatabine and S-(−)-anatabine can be calculated using the following formula. Ratio of R-(+)-anatabine to S-(−)-anatabine = Amount of R-(+)-anatabine by weight / Amount of S-(−)-anatabine by weight

[0048] When using solvates of R-(+)-anatabine and / or S-(−)-anatabine, the solvent is ignored in the calculation of the ratio.

[0049] The enantiomeric ratio can be determined by many methods including, but not limited to, chromatography, polarimetry of the rotation of polarization, nuclear magnetic resonance spectroscopy, derivatization of the compound using a chiral compound such as Mosher's acid followed by chromatography or nuclear magnetic resonance spectroscopy, high performance liquid chromatography (HPLC), or direct fractional crystallization of the racemate.

[0050] According to some embodiments of the present invention, the compound of formula I may be extracted from tobacco or may be chemically synthesized. The compound of formula I may be extracted from any of a variety of tobacco plants. As used herein, the terms "extracted" or "extracted" are intended to mean a substance obtained from a natural source or a process for obtaining a substance from a natural source. Alternatively, the compound of formula I may be chemically synthesized such that the compound is identical to the compound that occurs naturally.

[0051] In some embodiments, uses according to the invention are provided, wherein the formulation may comprise a concentrate of the compound of formula I.

[0052] According to the use of the present invention, the formulation may contain the compound of formula I in an amount of at least 99% by weight, at least 95% by weight, at least 90% by weight, at least 85% by weight, at least 80% by weight, at least 75% by weight, at least 70% by weight, at least 65% by weight, at least 60% by weight, at least 55% by weight, at least 50% by weight, at least 45% by weight, at least 40% by weight, at least 35% by weight, at least 30% by weight, at least 25% by weight, at least 20% by weight, at least 15% by weight, at least 10% by weight, at least 5% by weight, at least 2.5% by weight, at least 2.4% by weight, at least 2.3% by weight, at least 2.2% by weight, at least 2.1% by weight, at least 2% by weight, at least 1.9% by weight, at least 1.8% by weight, at least 1.7% by weight, at least 1.6% by weight, at least 1.5% by weight, at least 1.4% by weight, at least 1.3% by weight, at least 1.2% by weight, at least 1.1% by weight, at least 1% by weight, at least 0.9% by weight, at least 0.8% by weight, at least 0.7% by weight, at least 0.6% by weight, at least 0.5% by weight, at least 0.4% by weight, at least 0.3% by weight, at least 0.2% by weight, or at least 0.1% by weight of the formulation.

[0053] According to the use of the present invention, the formulation may contain the compound of formula I in an amount of 99 wt% or less, 95 wt% or less, 90 wt% or less, 85 wt% or less, 80 wt% or less, 75 wt% or less, 70 wt% or less, 65 wt% or less, 60 wt% or less, 55 wt% or less, 50 wt% or less, 45 wt% or less, 40 wt% or less, 35 wt% or less, 30 wt% or less, 25 wt% or less, 20 wt% or less, 15 wt% or less, 10 wt% or less, 5 wt% or less, 2.5 wt% or less, 2.4 wt% or less, 2.3 wt% or less, 2.2 wt% or less, 2.1 wt% or less, 2 wt% or less, 1.9 wt% or less, 1.8 wt% or less, 1.7 wt% or less, 1.6 wt% or less, 1.5 wt% or less, 1.4 wt% or less, 1.3 wt% or less, 1.2 wt% or less, 1.1 wt% or less, 1 wt% or less, 0.9 wt% or less, 0.8 wt% or less, 0.7 wt% or less, 0.6 wt% or less, 0.5 wt% or less, 0.4 wt% or less, 0.3 wt% or less, 0.2 wt% or less, or 0.1 wt% or less of the formulation.

[0054] According to the use of the present invention, the formulation may contain the compound of formula I in an amount of about 99 weight percent, about 95 percent, about 90 weight percent, about 85 weight percent, about 80 weight percent, about 75 weight percent, about 70 weight percent, about 65 weight percent, about 60 weight percent, about 55 weight percent, about 50 weight percent, about 45 weight percent, about 40 weight percent, about 35 weight percent, about 30 weight percent, about 25 weight percent, about 20 weight percent, about 15 weight percent, about 10 weight percent, about 5 weight percent, about 2.5 weight percent, about 2.4 weight percent, about 2.3 weight percent, about 2.2 weight percent, about 2.1 weight percent, about 2 weight percent, about 1.9 weight percent, about 1.8 weight percent, about 1.7 weight percent, about 1.6 weight percent, about 1.5 weight percent, about 1.4 weight percent, about 1.3 weight percent, about 1.2 weight percent, about 1.1 weight percent, about 1 weight percent, about 0.9 weight percent, about 0.8 weight percent, about 0.7 weight percent, about 0.6 weight percent, about 0.5 weight percent, about 0.4 weight percent, about 0.3 weight percent, about 0.2 weight percent, or about 0.1 weight percent of the formulation.

[0055] According to the use of the present invention, the formulation may contain the compound of formula I in an amount selected from 1 weight percent to 99 weight percent, 2.5 weight percent to 95 weight percent, 5 weight percent to 90 weight percent, 10 weight percent to 85 weight percent, 15 weight percent to 80 weight percent, 20 weight percent to 75 weight percent, 25 weight percent to 70 weight percent, 30 weight percent to 65 weight percent, 35 weight percent to 60 weight percent, 40 weight percent to 55 weight percent, 45 weight percent to 50 weight percent of the formulation. The amount of the compound of formula I may have any range from a given endpoint.

[0056] According to a further use of the present invention, the formulation may comprise the compound of formula I in an amount selected from 0.1% to 5% by weight, 0.15% to 4.9% by weight, 0.2% to 4.8% by weight, 0.25% to 4.7% by weight, 0.3% to 4.6% by weight, 0.35% to 4.5% by weight, 0.4% to 4.4% by weight, 0.45% to 4.3% by weight, 0.5% to 4.2% by weight, 0.55% to 4.1% by weight, 0.6% to 4.0% by weight, 0.65% to 3.9% by weight, 0.7% to 3.8% by weight, 0.75% to 3.7% by weight, 0.8% to 3.6% by weight, 0.85% to 3.5% by weight, 0.9% to 3.4% by weight, 0.95% to 3.3% by weight, 1% to 3.2% by weight, 1.15% to 3.1% by weight, 1.2% to 3.0% by weight, 1.25% to 2.9% by weight, 1.3% to 2.8% by weight, 1.35% to 2.7% by weight, 1.4% to 2.6% by weight, 1.45% to 2.5% by weight, 1.5% to 2.4% by weight, 1.6% to 2.3% by weight, 1.7% to 2.2% by weight, 1.8% to 2.1% by weight of the formulation. The amount of the compound of formula I can have any range from a given end point. According to some further embodiments of the present invention, the formulation may comprise the compound of formula I in an amount of 5%, 2%, 1%, or 0.5% by weight of the formulation.

[0057] In certain embodiments according to the present invention, the formulation may be a water-soluble concentrate, an emulsifiable concentrate, an emulsion, a microemulsion, an oily suspension concentrate, a flowable suspension, a water-dispersible granule, a water-soluble granule, a wettable powder, a water-soluble powder, a granule, an encapsulated granule, a fine granule, a macro granule, an aqueous suspoemulsion, a microencapsulated suspension, or a microgranule. Advantageously, the formulations of the present invention can be provided in different forms depending on the type of interaction required between the formulation and the agricultural crop or plant. Indeed, providing different forms of the formulations of the present invention allows the selection of the most suitable formulation for the intended use in order to ensure optimal biological efficiency depending on the targeted agricultural crop or plant.

[0058] According to the present invention, there is provided the use of a formulation that can be diluted with water. The formulations of the present invention may be diluted with water to produce the final product. The formulations of the present invention may be provided as pre-diluted, ready-to-use formulations, or may be concentrates intended for dilution prior to application. Advantageously, the pre-diluted ready-to-use formulations can be applied to plants or small agricultural crops. More advantageously, the formulations according to the present invention can be diluted immediately prior to application, allowing the user to adjust the dilution ratio of the formulation used for the agricultural crop or plant according to the intended purpose of the application.

[0059] According to the present invention, there is provided the use of a formulation, and the dilution ratio of the formulation to water can be selected from 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, 1:95, 1:100, 1:150, 1:200, 1:250, 1:300, 1:350, 1:400, 1:450, 1:500, 1:600, 1:700, 1:800, 1:900, or 1:1000.

[0060] According to the present invention, there is provided the use of a diluted formulation, the diluted formulation comprising the compound of formula I in an amount of at least 0.1 wt%, at least 0.2 wt%, at least 0.3 wt%, at least 0.4 wt%, at least 0.5 wt%, at least 0.6 wt%, at least 0.7 wt%, at least 0.8 wt%, at least 0.9 wt%, at least 1 wt%, at least 1.1 wt%, at least 1.2 wt%, at least 1.3 wt%, at least 1.4 wt%, at least 1.5 wt%, at least 1.6 wt%, at least 1.7 wt%, or at least 1.8 wt% of the diluted formulation.

[0061] According to the present invention, there is provided the use of a diluted formulation, the diluted formulation comprising the compound of formula I in an amount of 20 wt% or less, 19 wt% or less, 18 wt% or less, 17 wt% or less, 16 wt% or less, 15 wt% or less, 14 wt% or less, 13 wt% or less, 12 wt% or less, 11 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% or less, 7 wt% or less, 6 wt% or less, 5 wt% or less, 4 wt% or less, or 3 wt% or less of the diluted formulation.

[0062] According to the present invention, there is provided the use of a diluted formulation, the diluted formulation comprising the compound of formula I in an amount of from about 0.1 weight percent to about 20 weight percent, from about 0.2 weight percent to about 19 weight percent, from about 0.3 weight percent to about 18 weight percent, from about 0.4 weight percent to about 17 weight percent, from about 0.5 weight percent to about 16 weight percent, from about 0.6 weight percent to about 15 weight percent, from about 0.7 weight percent to about 14 weight percent, from about 0.8 weight percent to about 13 weight percent, from about 0.9 weight percent to about 12 weight percent, from about 1 weight percent to about 11 weight percent, from about 1.1 weight percent to about 10 weight percent, from about 1.2 weight percent to about 9 weight percent, from about 1.3 weight percent to about 8 weight percent, from about 1.4 weight percent to about 7 weight percent, from about 1.5 weight percent to about 6 weight percent, from about 1.6 weight percent to about 5 weight percent, from about 1.7 weight percent to about 4 weight percent, or from about 1.8 weight percent to about 3 weight percent of the diluted formulation.

[0063] In another embodiment of the present invention, there is provided the use of a diluted formulation, the diluted formulation comprising the compound of formula I in an amount of less than about 10 weight percent of the diluted formulation. In a further embodiment of the present invention, there is provided the use of a diluted formulation, the diluted formulation comprising the compound of formula I in an amount of about 5 weight percent of the diluted formulation. Surprisingly, at concentrations of less than 10 percent, the formulation has been shown not to cause damage to crops or plants.

[0064] In a further embodiment of the present invention, there is provided the use of a formulation, the formulation being applicable to agricultural crops or plants by coating, spraying, dipping, or perfusion. Advantageously, the choice of application method can target different families of hemipteran insects. For example, a method of deposition closer to the leaf with minimal air displacement may be more suitable for flying insects such as aphids.

[0065] In some embodiments of the present invention, there is provided the use of a formulation, which can be applied to agricultural crops or plants once, twice, three times, four times, five times, or six times within 24 hours.

[0066] According to the present invention, there is provided the use of a formulation, which can be applied to agricultural crops or plants daily, every two days, every five days, weekly, every two weeks (14 days), or monthly. Any application frequency can be combined with any of the aforementioned number of applications within 24 hours. According to the present invention, there is provided the use of a formulation, which can be applied routinely, regularly, or based on the evaluation after each application.

[0067] According to the present invention, there is provided the use of a formulation, which can be applied at the earliest detection time of insects or after the invasion becomes apparent. Alternatively, the formulation according to the present invention can be used preventively before the presence of insects is detected or before the invasion becomes apparent.

[0068] According to the present invention, there is provided the use of a formulation, which can be applied during a period of lower temperature in a day so that the formulation can be applied in the morning, evening, or night. Advantageously, applying the formulation according to the present invention during a period of lower temperature in a day has been found to reduce the possibility of evaporation of the formulation so that an effective concentration is maintained when the formulation is applied to agricultural crops or plants.

[0069] According to the present invention, there is provided the use of a formulation, which can be applied to agricultural crops or plants for at least 65 days, at least 60 days, at least 55 days, at least 50 days, at least 45 days, or at least 40 days, or at least 35 days, or at least 30 days, or at least 25 days, or at least 20 days.

[0070] According to the present invention, there is provided the use of a formulation, which can be applied to agricultural crops or plants for a period of 65 days or less, 60 days or less, 55 days or less, 50 days or less, 45 days or less, or 40 days or less.

[0071] Advantageously, the formulation according to the invention can be applied at a determined frequency for a period shorter than, equal to, or longer than the time of the life cycle of the hemipteran insects. More advantageously, the formulation according to the invention may be applied for a period equal to the period of the life cycle of the whiteflies at a determined frequency. Thus, the formulation according to the invention can be applied for a period of at least 40 days.

[0072] According to another aspect of the invention, there is provided a method for preventing hemipteran insects, the method comprising preparing a concentrated formulation comprising a compound of formula I, preferably anatabine, diluting the formulation with water to a predetermined dilution ratio or concentration, and applying the diluted formulation to agricultural crops or plants, preferably by spraying.

[0073] The invention is further illustrated by, but not limited to, the following examples.

[0074] Example 1: Comparative study on the insect repellent effects of anatabine and DEET. The insect repellent effect of anatabine against adult whiteflies was evaluated in a comparative study comparing the insect repellent effect of anatabine with that of the standard synthetic insecticide DEET and a placebo control in which the leaves were treated with the solvent only.

[0075] For the purpose of the study, a tube test assay was carried out using solutions of anatabine and DEET prepared according to Table 1. [Table 1]

[0076] For the purpose of the study, leaf discs were immersed in the prepared solutions for a total of 30 seconds and dried at room temperature for 10 minutes. Thereafter, the leaf discs were deposited at the bottom of a glass tube and at least 20 adult whiteflies were introduced into the tube. The insect repellent effects of both anatabine and DEET were measured at different time points after 1, 3, and 6 hours. The insect repellent effects of both anatabine and DEET are shown in Table 2 and were calculated by counting the number of whiteflies on the leaves and comparing them with the placebo control using Abbott's formula. Insect - repellent effect = 1 - (Number of aphids on the leaves of the "placebo control" / Number of aphids on the leaves of the "treated")×100.

[0077] The placebo control defines a reference value, and based on that, the insect - repellent effect is calculated according to the above formula.

Table 2

[0078] The comparative study found that while the insect - repellent effect of anatabine was over 80% throughout the 6 - hour evaluation period, the insect - repellent effect of DEET decreased to approximately 40% (from about 50%) after 6 hours (Figure 1).

[0079] Example 2: Study on the insect - repellent effect of anatabine with respect to its concentration. The insect - repellent effect of anatabine against adult aphids was evaluated in a study aimed at optimizing the concentration of anatabine.

[0080] For the purpose of the study, a tube - test assay was conducted using three different concentrations of anatabine solutions prepared according to Table 3.

[0081] The placebo control defines a reference value, and then the insect - repellent effect is calculated based on it.

Table 3

[0082] For the purpose of the study, leaf disks were immersed in the prepared solutions for a total of 30 seconds and dried at room temperature for 10 minutes. Then, the leaf disks were deposited at the bottom of a glass tube, and at least 20 adult aphids were introduced into the tube. The insect - repellent effect of anatabine at three different concentrations was measured at different time points: 1 hour later, 3 hours later, and 6 hours later. The insect - repellent effect of anatabine at different concentrations is shown in Table 4.

Table 4

[0083] The insect - repellent effect of anatabine remained 100 percent regardless of the concentration of anatabine (Figure 2), although it was also observed that the leaves were not damaged by anatabine at concentrations of 5 percent or less.

[0084] Example 3: Comparative study on the insect - repellent effects of anatabine, DEET, and methyl benzoate. The insect - repellent effect of anatabine against adult aphids was evaluated in a comparative study aimed at comparing the insect - repellent effect of anatabine with DEET and methyl benzoate. DEET is a standard synthetic insecticide, and methyl benzoate is a plant - derived organic compound that has been reported to have insect - repellent and insecticidal properties, especially against aphids.

[0085] For the purpose of the study, a tube - test assay was carried out using solutions of anatabine, DEET, and methyl benzoate prepared according to Table 5. A concentration of 5 percent was selected based on the results of Example 2. The insect - repellent effects of anatabine, DEET, and methyl benzoate were evaluated with respect to a placebo control. In the placebo control, the leaves were treated with the solvent only. The placebo control defined a reference value against which the insect - repellent effect was then calculated.

Table 5

[0086] For the purpose of the study, leaf disks were immersed in the prepared solutions for a total of 30 seconds and dried at room temperature for 10 minutes. Then, the leaf disks were deposited at the bottom of a glass tube, and at least 20 adult aphids were introduced into the tube. The insect - repellent effects of anatabine, DEET, and methyl benzoate were measured at different time points after 6 hours, 24 hours, and 48 hours. The insect - repellent effects of anatabine, DEET, and methyl benzoate are shown in Table 6.

Table 6

[0087] The insect - repellent effect of anatabine remained 100% throughout the 48 - hour evaluation period. In contrast, the insect - repellent effect of methyl benzoate decreased by nearly 25% by the end of the 48 - hour evaluation period (Figure 3), and the insect - repellent effect of DEET decreased to 93%.

[0088] Furthermore, the presence and amount of eggs on the leaves after 48 hours were visually evaluated, and the results are shown in Table 7.

Table 7

[0089] Regardless of the insect - repellent used, no eggs were observed on the leaves treated with the insect - repellent, but they were clearly visible on the untreated leaves.

[0090] This study also evaluated the mortality rate of aphids over the evaluation period. The results are shown in Table 8.

Table 8

[0091] When the leaves were treated with placebo, the mortality rate of aphids remained less than 10% throughout the evaluation period. In contrast, when the leaves were treated with an insect - repellent to prevent aphid feeding, the aphids began to die within a day. By the end of the 48 - hour evaluation period, the aphid mortality rate for the treated leaves reached approximately 80 percent, as shown in Figure 4.

[0092] To better understand the mortality rate, a population of aphids without access to the leaves over the 48 - hour evaluation period was also observed. Without access to a food source, the aphids began to die after just 24 hours and reached a 60% mortality rate after 48 hours.

[0093] Furthermore, DEET and methyl benzoate were observed to cause severe damage to the leaves over the 48 - hour evaluation period. In contrast, anatabine caused no damage over the evaluation period, as shown in Table 9.

Table 9

[0094] Example 4: Comparative study on the insect repellent effects of anatabine and DEET. The insect repellent effect of anatabine against adult bed bugs was evaluated in a further comparative study, which introduced additional bed bugs and newly treated leaves at the set evaluation time points. For the purposes of the study, as shown in Table 10, assays were conducted using solutions of anatabine and DEET at a concentration of 5 percent together with a placebo solution in a tube test assay. The placebo control defined a baseline value against which the insect repellent effect was then calculated.

Table 10

[0095] Bed bugs were introduced into the tubes at the invasion time points of 48 hours, 72 hours, and 120 hours, and the insect repellent effect was measured at the evaluation time points of 3 hours later, 51 hours, 75 hours, and 123 hours. The results are shown in Table 11.

Table 11

[0096] Anatabine was shown to have 100% efficacy 48 hours after treatment and also to have an insect repellent effect of over 80% for the remainder of the 5-day evaluation period (up to 123 hours). In contrast, the efficacy of DEET decreased by almost 50% after the first 48 hours and remained below 50% for the remainder of the 5-day evaluation period (up to 123 hours) (Figure 5).

[0097] The presence of eggs was visually monitored 24 hours from each invasion time point. The results are shown in Table 12.

Table 12

[0098] The number of eggs observed on leaves treated with anatabine remained low throughout the 6-day (144-hour) evaluation period. In contrast, eggs were observed on leaves treated with DEET after just 3 days (72 hours), and by the end of the 6-day (144-hour) evaluation period, the number of eggs was close to that on leaves treated with placebo.

[0099] Although the invention is defined in the claims, a non-exhaustive list of non-limiting aspects is provided below. Any one or more of the features of these aspects may be combined with any one or more of the features of any other aspect described herein.

[0100] Aspect 1: Use of a formulation for preventing or reducing infestation by Hemiptera insects, wherein the formulation comprises a compound of formula I,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0101] For the purposes of this specification and the appended claims, unless otherwise indicated, all numbers expressing amounts, quantities, percentages, ratios, etc. are to be understood as being modified in all instances by the term "about". Also, all ranges include the disclosed maximum and minimum points, and any intermediate ranges therebetween, whether or not specifically enumerated herein. Thus, in this context, number A is understood as A ± 5% of A. Within this context, number A may be considered to include numerical values within the normal standard error of measurement of the property being modified by number A. Number A may deviate by the percentages recited above in some instances as used in the appended claims, provided that the amount by which A deviates does not substantially affect the basic and novel characteristics of the claimed invention. Also, all ranges include the disclosed maximum and minimum points, and any intermediate ranges therebetween, whether or not specifically enumerated herein.

Claims

1. The use of formulations to prevent or reduce the invasion of hemipteran insects, The aforementioned hemipteran insect is an insect that feeds on tree sap, and the formulation contains the compound of formula I. 【Chemistry 1】 During the ceremony, R is hydrogen or C 1 -C 5 Represents alkyl, 【Chemistry 2】 The symbol is used to represent a single bond or a double bond.

2. The use according to claim 1, wherein the aforementioned hemipteran insect is a whitefly.

3. The use according to claim 1 for preventing or reducing the invasion of hemipteran insects from agricultural crops or plants.

4. The use according to claim 3, wherein the agricultural crop or plant is vulnerable to whitefly invasion.

5. The use according to claim 3, wherein the agricultural crop or plant is selected from tobacco plants, cereal plants, legumes, oil plants, cucurbits, fiber plants, vegetables, fruits, flowers, and ornamental plants.

6. The use according to claim 3, wherein the agricultural crop or plant is a tobacco plant.

7. The use according to claim 3, wherein the agricultural crop or plant is cultivated in a greenhouse.

8. The use according to claim 1, wherein the compound of formula I is selected from anatabine, S-(-)anatabine, R-(+)anatabine, a mixture of S-(-)anatabine and R-(+)anatabine, or a racemic mixture of S-(-)anatabine and R-(+)anatabine.

9. The use according to claim 1, wherein the formulation comprises a compound concentrate of formula I.

10. The use according to claim 1, wherein the formulation contains at least 99 weight percent, at least 95 weight percent, at least 90 weight percent, at least 85 weight percent, at least 80 weight percent, at least 75 weight percent, at least 70 weight percent, at least 65 weight percent, at least 60 weight percent, at least 55 weight percent, at least 50 weight percent, at least 45 weight percent, at least 40 weight percent, at least 35 weight percent, at least 30 weight percent, at least 25 weight percent, at least 20 weight percent, at least 15 weight percent, at least 10 weight percent, at least 5 weight percent, at least 2.5 weight percent, or at least 1 weight percent of the compound of formula I.

11. The use according to claim 1, wherein the formulation is diluted with water.

12. The use according to claim 11, wherein the amount of the compound of formula I is 0.1 to 20 weight percent, 0.2 to 19 weight percent, 0.3 to 18 weight percent, 0.4 to 17 weight percent, 0.5 to 16 weight percent, 0.6 to 15 weight percent, 0.7 to 14 weight percent, 0.8 to 13 weight percent, 0.9 to 12 weight percent, 1 to 11 weight percent, 1.1 to 10 weight percent, 1.2 to 9 weight percent, 1.3 to 8 weight percent, 1.4 to 7 weight percent, 1.5 to 6 weight percent, 1.6 to 5 weight percent, 1.7 to 4 weight percent, or 1.8 to 3 weight percent of the diluted formulation.

13. The use according to claim 1, wherein the formulation is applied to agricultural crops or plants by coating, spraying, dipping, or drenching.

14. A method for preventing Hemiptera insects, wherein the method is a. Compounds of formula I 【Transformation 3】 (In the formula, R is hydrogen or C 1 -C 5 Represents alkyl, 【Chemistry 4】 The process involves preparing a concentrated formulation containing anatabine, preferably with the notation representing a single or double bond. b. Diluting the preparation with water to a predetermined dilution ratio or concentration, c. A method comprising applying the diluted formulation to agricultural crops or plants, preferably by spraying.

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