Organosulfur compounds as plant biostimulants.

Abstract

The present disclosure relates to organic sulfur-containing compositions, particularly di-n-propyl thiosulfonate (PTSO) and di-n-propyl thiosulfinate (PTS), dimethyl thiosulfonate, and diphenyl thiosulfonate. Such compositions are useful as biostimulants for plants. In particular, such compositions may result in improved nutrient utilization efficiency, improved resistance to abiotic stress, and / or improved quality attributes. Compositions comprising said organic sulfur compounds are also provided for agricultural use.

Description

[Technical field]

[0001] The present disclosure relates to organic sulfur-containing compositions, particularly di-n-propyl thiosulfonate (PTSO) and di-n-propyl thiosulfinate (PTS), dimethyl thiosulfonate, diphenyl thiosulfonate, and di-n-propyl disulfide. Such compositions are useful as biostimulants for plants. In particular, such compositions may result in improved nutrient utilization efficiency, improved abiotic stress tolerance, and / or improved quality characteristics. Compositions comprising the organic sulfur compounds are also provided for agricultural use. [Background technology]

[0002] Plants are ideally grown under optimum conditions to obtain desired characteristics, such as high yield or high quality.

[0003] Growth conditions can be optimized in terms of root substrate, fertilization, watering, drainage, light supply, temperature, air movement, air humidity and pest control. During crop and plant production in horticulture, most of the aforementioned growth conditions are maintained under strict control. However, there are situations where the control system is unable to maintain the optimal parameters under precise control, e.g., sudden changes in weather conditions. Examples include alternating periods of sunny and cloudy weather, resulting in short periods of sunshine and high temperatures, or vice versa. Sudden temperature changes, e.g., cold nights with increased heating and decreased humidity, result in high evaporation rates by plants. Long periods of cloudy weather result in low photosynthetic rates and high humidity. Long periods of sunlight generate excess heat and low humidity, leading to high evaporation by plants.

[0004] Even during favorable growing seasons, growing conditions may occur where plants are unable to take up sufficient water and nutrients, causing them to grow more slowly than if there was an excess of nutrients and water.

[0005] Plant nutrients are chemical elements and compounds necessary for plant growth and / or plant metabolism. For example, vascular plants require the following elements for growth and development: carbon (C), hydrogen (H), oxygen (O), nitrogen (N), phosphorus (P), potassium (K), sulfur (S), calcium (Ca), magnesium (Mg), boron (B), chlorine (Cl), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), nickel (Ni) and zinc (Zn). Many vascular plants also require silicate (Si).

[0006] In agriculture, nitrogen (N), phosphorus (P), potassium (K), sulfur (S), calcium (Ca), magnesium (Mg), boron (B), chlorine (Cl), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), nickel (Ni), zinc (Zn) and silicate (Si) are commonly provided as fertilizers. These elements can be provided as relatively pure salts or as complex organic matter, such as compost, plant waste, or (partially) decomposed animal faeces, such as cows, chickens and pigs, and are usually provided in excess to prevent nutrient limitation.

[0007] It is clear that during open-field agriculture plants are grown under more variable conditions than in greenhouses. Temperature, sunshine and root support / air humidity, but also wind, frost, salinity, flooding and drought are important abiotic factors. The conditions described result in abiotic stress for the crop, resulting in reduced yields and deterioration of crop quality parameters. Abiotic stress is defined as the adverse effect of abiotic factors on living organisms in a particular environment. It is clear that some crops and ornamental plants are less tolerant to abiotic stresses, even under controlled greenhouse conditions. This strongly depends on the species, subspecies hybrid, variant or cultivar.

[0008] In recent years, the use of plant biostimulants has been attracting attention as a means to improve plant characteristics. According to Du Jardin, P (2012) “The science of plant biostimulants-a bibliographic analysis. Ad hoc Study Report to the European Commission DG ENTR;2012”, a plant biostimulant is any substance or microorganism that is applied to plants with the aim of increasing nutrient efficiency, abiotic stress resistance and / or crop quality traits, regardless of nutrient content. Biostimulants have the ability to modify physiological processes of plants in ways that potentially benefit growth, development and / or stress responses. Summary of the Invention [Problem to be solved by the invention]

[0009] One object of the present disclosure is to provide plant biostimulants suitable for use in agriculture. [Means for solving the problem]

[0010] The present disclosure provides the following preferred embodiments.

[0011] 1. The use of a compound according to formula I below or a composition comprising a compound according to formula I below as a biostimulant for plants, wherein formula I is [ka] and where n is 2; wherein one X is -S- and the other X is -S-, -S(O)-, and -S(O) 2 - selected from the group consisting of; Also, R 1 and R 2 are each independently selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl, preferably, wherein the compound according to formula I is di-n-propylthiosulfonate (PTSO), di-n-propyl disulfide, dimethylthiosulfonate, or diphenylthiosulfonate.

[0012] 2. A method comprising providing a plant with a compound according to formula I below or a composition comprising a compound according to formula I below. [ka] where n is 2; wherein one X is -S- and the other X is -S-, -S(O)-, and -S(O) 2 - selected from the group consisting of; Also, R 1 and R 2are independently selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl, preferably wherein the compound according to formula I is di-n-propylthiosulfonate (PTSO), di-n-propyl disulfide, dimethylthiosulfonate, or diphenylthiosulfonate. Preferably, the method is for increasing the plant growth rate, plant development, plant yield, and / or plant harvest. The method may also increase plant vigor. Preferably, the method is for increasing nutrient utilization efficiency, for increasing the plant resistance to biotic stress, for improving the quality attributes of the plant, and / or for increasing the availability of nutrients retained in the soil or in the rhizosphere for the plant.

[0013] 3.R 1 and R 2 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted aryl.

[0014] 4.R 1 and R 2 are independent of each other, C 1~6 alkyl and phenyl; wherein the phenyl group is selected from C 1~3 Alkyl or C 1~3 optionally substituted with alkoxy; Here, preferably, R 1 and R 2 are independently selected from methyl, ethyl, n-propyl, n-butyl, phenyl, p-tolyl and 4-methoxyphenyl; The use or method according to any one of the embodiments of items 1 to 3.

[0015] 5.R 1 and R 2The use or method according to any one of the embodiments of items 1 to 4, wherein

[0016] 6. The compound according to formula I is selected from the group consisting of di-n-propyl thiosulfonate (PTSO), S-methyl methane thiosulfonate, S-phenyl benzene thiosulfonate, di-n-propyl thiosulfinate (PTS), methyl methane thiosulfinate, butyl butane thiosulfinate, methyl propene thiosulfinate, di-n-propyl disulfide, dimethyl disulfide, diethyl disulfide, di-n-butyl disulfide, diphenyl disulfide, di-p-tolyl disulfide, and bis(4-methoxyphenyl) disulfide; Preferably, the compound according to formula I is di-n-propylthiosulfonate (PTSO) or di-n-propylthiosulfinate (PTS), The use or method according to any one of the embodiments of items 1 to 5.

[0017] 7. The use or method according to any one of the embodiments 1 to 6, wherein at least 60% by weight, preferably at least 80% by weight, more preferably at least 95% by weight, of the organosulfur compounds in the composition are selected from compounds according to formula I.

[0018] 8. When the composition comprises diallyl thiosulfinate, the weight ratio of the compound or the compound according to formula I to diallyl thiosulfinate is greater than 0.1, preferably greater than 1, and wherein, when the composition comprises diallyl disulfide, the weight ratio of the compound or the compound according to formula I to diallyl disulfide is greater than 0.1, preferably greater than 1; The use or method according to any one of the embodiments of items 1 to 7.

[0019] 9. The use as a biostimulant, a) nutrient use efficiency, b) resistance to abiotic stress, c) quality attributes, and d) availability of confined nutrients held in the soil or in the rhizosphere. The use or method according to any one of the embodiments of items 1 to 8, which improves one or more of the following:

[0020] 10. The use or method according to any one of the preceding embodiments, wherein the composition is essentially free of diallyl thiosulfinate.

[0021] 11. The use or method according to any one of the preceding embodiments, wherein the composition is essentially free of diallyl disulfide.

[0022] 12. The use, method or composition according to any one of the preceding embodiments, wherein the composition comprises at least 0.5 mg / L of a compound according to formula I.

[0023] 13. The use or method according to any one of the embodiments of paragraphs 1 to 12, wherein the plant belongs to the clade Embryophyta or the clade Angiospermae.

[0024] 14. The use or method according to any one of the preceding embodiments, wherein the plant is selected from the group consisting of Phalaenopsis, Chrysanthemum, Solanum lycopersicum and Lactuca sativa.

[0025] 15. The use or method according to any one of the preceding embodiments, wherein the composition further comprises a fertilizer, an insecticide, a wetting agent, an antimicrobial compound, a bactericide, a chelating compound, an aromatic compound, and / or an additional biostimulant.

[0026] 16. The use or method according to any one of the preceding embodiments, wherein the compound or the composition is applied directly to the plant, to the seeds of the plant, or to the soil of the plant or the seeds.

[0027] 17. The use or method according to embodiment 16, wherein the compound or the composition is applied via drip irrigation.

[0028] 18. A method comprising providing to a plant, preferably via irrigation, a composition comprising a compound according to formula I: [ka] where n is 2; wherein one X is -S- and the other X is -S-, -S(O)-, and -S(O) 2 - selected from the group consisting of; Also, R 1 and R 2 are independently selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl, preferably wherein the compound according to formula I is di-n-propylthiosulfonate (PTSO), di-n-propyl disulfide, dimethylthiosulfonate, or diphenylthiosulfonate; wherein the composition is provided to the plant at least 6 times during the crop cycle of the plant, and / or the composition is provided to the plant every 4 to 21 days during the crop cycle of the plant.

[0029] 19. The use, method or composition according to any one of the preceding embodiments, wherein the composition comprises an emulsifier.

[0030] 20. The use, method or composition according to any one of the preceding embodiments, wherein the composition comprises an amino acid-based biostimulant.

[0031] 21. The use, method or composition according to any one of the preceding embodiments, wherein the composition comprises free amino acids and peptides.

[0032] 22. The use or method according to any one of the preceding embodiments, wherein the use or method further comprises providing the plant with an amino acid-based biostimulant.

[0033] 23. The use or method according to any one of the preceding embodiments, wherein the amino acid-based biostimulant comprises 8-15% (w / w) free amino acids and 45-55% attached amino acids.

[0034] 24. The use or method according to any one of embodiments 22 to 23, wherein the composition comprising a compound according to formula I and an amino acid-based biostimulant are provided within the same crop cycle.

[0035] 25. The use or method according to any one of the embodiments of paragraphs 22 to 24, wherein the composition comprising a compound according to formula I is provided at least 24 hours before or after the amino acid-based biostimulant is provided to the plant. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] The present disclosure provides organosulfur compounds for use as plant biostimulants and methods for increasing plant growth rate, plant development, plant yield and harvest, and other plant characteristics described herein.

[0037] In some embodiments, the organosulfur compound is a compound according to Formula I: [ka] where n is 2; wherein one X is -S- and the other X is -S-, -S(O)-, and -S(O) 2 - selected from the group consisting of; Also, R 1 and R 2 are each independently selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, and optionally substituted heterocycloalkyl.

[0038] In a preferred embodiment, the compound according to formula I is [ka] No.

[0039] In a preferred embodiment, the compound according to formula I is [ka] No.

[0040] The compounds according to Formula I are referred to herein as "organosulfur compounds" or "biostimulant organosulfur compounds."

[0041] Preferably, R 1 and R 2 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted aryl. More preferably, R 1 and R 2 are independent of each other, C 1~6 alkyl and phenyl, wherein the phenyl group is selected from C 1~3 Alkyl or C 1~3 It is preferably substituted with alkoxy. Most preferably, R 1 and R 2are independently selected from methyl, ethyl, n-propyl, n-butyl, phenyl, p-tolyl and 4-methoxyphenyl.

[0042] In a preferred embodiment, R 1 and R 2 are identical.

[0043] In a preferred embodiment, each X is -S-. In a preferred embodiment, one X is -S- and the other X is -S(O)-. In another preferred embodiment, one X is -S- and the other X is -S(O)-. 2 -It is.

[0044] Preferably, the compound of formula I is selected from the group consisting of di-n-propylthiosulfonate (PTSO), S-methylmethanethiosulfonate, S-phenylbenzenethiosulfonate, di-n-propylthiosulfinate (PTS), methylmethanethiosulfinate, butylbutanethiosulfinate, methylpropenethiosulfinate, di-n-propyldisulfide, dimethyldisulfide, diethyldisulfide, di-n-butyldisulfide, diphenyldisulfide, di-p-tolyldisulfide, and bis(4-methoxyphenyl)disulfide. More preferably, the compound according to formula I is di-n-propylthiosulfonate (PTSO) or di-n-propylthiosulfinate (PTS).

[0045] In a preferred embodiment of formula I, n is 2 and one X is -S- and the other X is -S(O) 2 - and; and R 1 and R 2 are independent of each other, C 1~6 It is selected from alkyl and phenyl, and is preferably selected from the group consisting of methyl, phenyl and n-propyl.

[0046] In a preferred embodiment, the compound of formula I is propyl-propane thiosulfonate (PTSO), also referred to as di-n-propylthiosulfonate. PTSO has the following structure: [ka]

[0047] In a preferred embodiment, the compound of formula I is propyl-propane-thiosulfinate (PTS), also referred to as di-n-propyl thiosulfinate. PTS has the following structure: [ka]

[0048] In a preferred embodiment, the compound of formula I is dimethylthiosulfonate.

[0049] In a preferred embodiment, the compound of formula I is a diphenylthiosulfonate.

[0050] PTSO and PTS are preferred. The compounds of the present disclosure may be used together or individually as biostimulants and in the methods described herein.

[0051] As used herein, "alkyl" refers to a saturated aliphatic hydrocarbyl group. Unless otherwise specified, the alkyl group can be linear or branched. Preferably, the alkyl group is linear. As used herein, the alkyl group can be substituted or unsubstituted. Preferably, the alkyl group is unsubstituted. In a preferred embodiment, in formula I, the alkyl group is selected from the group consisting of C 1~6 Alkyl, more preferably C 1~4 Alkyl.

[0052] As used herein, "aryl" refers to an aromatic hydrocarbon ring system containing 6 to 24 carbon atoms, more preferably 6 to 12 carbon atoms, and may include monocyclic and polycyclic structures. When the aryl group is a polycyclic structure, it is preferably a bicyclic structure. Optionally, the aryl group may be substituted with one or more substituents as further specified herein. Preferably, the aryl group is substituted with a methyl or methoxy group. Examples of the aryl group are phenyl and naphthyl. Most preferably, the aryl group is phenyl.

[0053] As used herein, "alkenyl" refers to an unsaturated aliphatic hydrocarbyl group containing one or more carbon-carbon double bonds. Unless otherwise specified, an alkenyl group can be linear or branched. Preferably, an alkenyl group is linear. As used herein, an alkenyl group can be substituted or unsubstituted. Preferably, an alkenyl group is unsubstituted. In a preferred embodiment, in formula I, alkenyl is C 2~6 Alkenyl, more preferably C 2~4 Alkenyl.

[0054] As used herein, "alkynyl" refers to an unsaturated aliphatic hydrocarbyl group containing one or more carbon-carbon triple bonds. Unless otherwise specified, an alkynyl group can be linear or branched. Preferably, an alkynyl group is linear. As used herein, an alkynyl group can be substituted or unsubstituted. Preferably, an alkynyl group is unsubstituted. In a preferred embodiment, in formula I, an alkynyl group is selected from the group consisting of C 2~6 Alkynyl, more preferably C 2~4 Alkynyl.

[0055] As used herein, "cycloalkyl" refers to a cyclic saturated aliphatic hydrocarbyl group. The cycloalkyl group can be substituted or unsubstituted. Preferably, the cycloalkyl group is unsubstituted. In a preferred embodiment, in formula I, the cycloalkyl is C 3~6 Cycloalkyl, more preferably C 3~5 Cycloalkyl.

[0056] As used herein, "heteroalkyl" refers to a saturated aliphatic hydrocarbyl group containing one or more heteroatoms. Unless otherwise specified, the heteroalkyl group can be linear or branched. Preferably, the heteroalkyl group is linear. As used herein, the heteroalkyl group can be substituted or unsubstituted. Preferably, the heteroalkyl group is unsubstituted. In a preferred embodiment, in formula I, the heteroalkyl group is selected from the group consisting of C 1~6 Heteroalkyl, more preferably C 1~4 Heteroalkyl. Preferably, the heteroalkyl group contains one or more heteroatoms selected from the group consisting of O, N and S, more preferably, the heteroalkyl group contains up to two heteroatoms, most preferably one heteroatom. Examples of preferred heteroalkyl groups include alkoxy groups (e.g., methoxy and ethoxy groups) and ethers.

[0057] As used herein, "heterocycloalkyl" refers to a cyclic saturated aliphatic hydrocarbyl group containing one or more heteroatoms. The heterocycloalkyl group can be substituted or unsubstituted. Preferably, the heterocycloalkyl group can be unsubstituted. In a preferred embodiment, in formula I, the heterocycloalkyl is C 1~5 Heterocycloalkyl, more preferably C 2~4Preferably, the heterocycloalkyl group is a 5- or 6-membered ring structure containing up to two heteroatoms, more preferably one heteroatom. Preferably, the heterocycloalkyl group contains a heteroatom selected from the group consisting of O, N, and S.

[0058] As used herein, "heteroaryl" refers to an aromatic ring system containing one or more heteroatoms. Preferably, a heteroaryl group contains at least two carbon atoms (i.e., at least C 2 ) and one or more heteroatoms N, O or S. Preferably, the heteroaryl group contains at most 5 carbon atoms. Preferably, the heteroaryl group contains up to 2 heteroatoms selected from the group consisting of N, O and S. In a preferred embodiment, the heteroaryl group is a 5- or 6-membered ring structure. Optionally, the heteroaryl group may be substituted by one or more substituents as further specified herein. Preferably, the heteroaryl group is unsubstituted. Examples of suitable heteroaryl groups include pyridinyl, quinolinyl, pyrimidinyl, pyrazinyl, pyrazolyl, imidazolyl, thiazolyl, pyrrolyl, furanyl, triazolyl, benzofuranyl, indolyl, purinyl, benzoxazolyl, thienyl, phosphoryl and oxazolyl.

[0059] As used herein, "substituted" indicates that a group contains one or more substituents. Preferably, the substituents are, independently of each other, halogen, C 1~3 Alkyl, -C(O)OH, -C(O)NH 2 , -OH, =O, C 1~3 Alkoxy, -NH 2 , -NH-C 1~3 Alkyl, -NHC(O)-C 1~3 Alkyl, -NO 2 , -SO 3 H, and CF 3Preferably, the halogen is selected from the group consisting of -Cl, -F, -Br, and -I. In preferred embodiments, groups as disclosed herein contain no more than three substituents, more preferably no more than two substituents, and most preferably no more than one substituent.

[0060] A number of organosulfur compounds have been identified in extracts from plants belonging to the Allium family. Propyl-propane-thiosufinate (PTS) is a naturally occurring compound found in plants belonging to the Allium family, particularly Allium cepa (onion), Allium ampeloprasum (leek), Allium schoenoprasum (chive), and Allium chinense (Chinese onion). Propiin is hydrolyzed by alliinase to propylsulfenic acid, which condenses with loss of water to produce PTS (J. Chromatogr. A 1112 (2006) 3-22). Further reaction with PTS results in the production of PTSO. One of the best plant sources for PTS is Allium schoenoprasum (chive) (see Table 3 in Rose et al. Nat. Prod. Rep., 2005, 22, 351-368). In contrast to PTS, PTSO is not present in onion. PTSO has been described in the literature as "derived from Allium", but the inventors are not aware of any literature describing the measurement of PTSO from onion extracts. Example 10 demonstrates that PTSO is not present at detectable limits in onion oil, onion extract, garlic oil or garlic extract.

[0061] Allium sativum (garlic) has been reported to contain undetectable levels of PTS, but significant amounts of allicin (i.e., diallyl thiosulfinate) cannot be detected in extracts of Allium cepa (onion), Allium ascalonicum (shallot), or Allium schoenoprasum (chive) (Rose et al.). Allicin is produced as a major compound from the non-proteinogenic amino acid alliin (S-allyl cysteine ​​sulfoxide) in a reaction catalyzed by the enzyme alliinase, together with minor amounts of methyl allyl thiosulfinate, in response to damage to raw garlic tissue (Molecule, 19, 2014, 12591-12618 and J. Chromatogr. A 1112 (2006) 3-22). Allicin is unstable and is readily converted to a series of other sulfur-containing compounds, such as diallyl disulfide. Because allicin is an unstable compound, its use in agriculture is limited (Fujisawa et al (2008) J Agric Food Chem:56 (11):4229-4235).

[0062] The compounds can be extracted from natural sources or synthetically produced. Both compounds PTS and PTSO are also commercially available. In some embodiments, the compounds are obtained from natural sources, such as plants. Compounds can be extracted from plant materials in a variety of ways. The appropriate method depends on the chemical properties of the compound. For example, extraction can begin with a non-polar solvent, followed by extraction with solvents of increasing polarity. Alternatively, the plant compounds can be extracted in alcohol.

[0063] The present disclosure provides compositions comprising the biostimulatory organosulfur compounds disclosed herein (i.e., according to Formula I).

[0064] In some embodiments, the composition comprises at least 40%, preferably at least 50%, of one or more organosulfur compounds. Preferably, the composition comprises at least 40%, preferably at least 50%, of PTSO. In some embodiments, the composition further comprises preferably less than 20%, more preferably less than 10%, of PTS. Such compositions are also referred to herein as "concentrated solutions of organosulfur compounds."

[0065] In some embodiments, at least 60% by weight, preferably at least 80% by weight, more preferably at least 95% by weight, of the organosulfur compounds of the compositions disclosed herein are selected from compounds according to Formula I. In some embodiments, at least 60% by weight, preferably at least 80% by weight, more preferably at least 95% by weight, of the organosulfur compounds of the compositions disclosed herein are selected from PTSO and PTS.

[0066] The concentrated compositions are typically diluted 1:100 to 1:100,000 prior to use to form working solutions. It will be understood that in this specification, "working" refers to the composition, e.g., solution, that can be applied to plants, and does not refer to other concentrations that relate to non-working embodiments.

[0067] A suitable composition comprises at least 0.5 mg / L of the organosulfur compound disclosed herein. Preferably, the composition comprises at least 0.5 mg / L, more preferably at least 1 mg / L of the organosulfur compound, particularly PTSO. In some embodiments, the working solution comprises at least 10 mg / L of the organosulfur compound, particularly PTSO.

[0068] Preferably, the composition contains at least 1 μmol / L, more preferably at least 10 μmol / L, of the organosulfur compound, particularly PTSO. In some embodiments, the working solution contains at least 35 μmol / L of the organosulfur compound, particularly at least 35 μmol / L of PTSO, more particularly at least 50 μmol / L of PTSO.

[0069] Preferably, the composition, especially when used as a working solution, contains 10 mmol / L or less, more preferably 8 mmol / L or less of organosulfur compounds, especially PTSO. In some embodiments, the working solution contains 6 mmol / L or less of organosulfur compounds, especially PTSO. In a preferred embodiment, the working solution contains 3.5 mmol / L or less of PTSO, more preferably 1.5 mmol / L or less of PTSO.

[0070] In a preferred embodiment, the compound according to formula I is applied to the plant (i.e., working solution) at a concentration of 200 mg / L or less, preferably 150 mg / L or less, more preferably 100 mg / L or less. In a preferred embodiment, the composition applied to the plant comprises 50 mg / L or less. In some embodiments, the composition applied to the plant comprises 0.5 to 150 mg / L, preferably 0.5 to 100 mg / L. In some embodiments, the composition applied to the plant comprises 0.5 to 50 mg / L. In some embodiments, the composition applied to the plant comprises 1 to 50 mg / L. In some embodiments, the composition applied to the plant comprises 1 to 10 mg / L.

[0071] Those skilled in the art will recognize that the amount of PTSO required will depend on the size of the crop, with larger crops generally requiring larger amounts. In an exemplary embodiment, the average plant density may be, for example, 30,000 plants / ha, and 10,000 liters of PTSO solution / ha may be applied.

[0072] In some embodiments, the composition comprises an additional organosulfur compound, which may or may not act as a biostimulant. Preferably, at least 60% by weight of the organosulfur compounds in the composition are selected from compounds according to formula I.

[0073] In some embodiments, when the composition includes diallyl thiosulfinate, the weight ratio of the compound or compounds according to formula I to the diallyl thiosulfinate is greater than 0.1, preferably greater than 1, and more preferably at least 10: 1. It will be understood that when the composition includes two or more compounds of formula I, the weights of all these compounds reach the above ratio compared to the weight of the diallyl thiosulfinate.

[0074] In some embodiments, when the composition includes diallyl disulfide, the weight ratio of the compound or compounds according to formula I to diallyl disulfide is greater than 0.1, preferably greater than 1, more preferably at least 10:1. It will be understood that when the composition includes more than one compound of formula I, the weights of all of these compounds will be in the above ratios compared to the weight of diallyl disulfide.

[0075] Preferably, such compositions are substantially free of diallyl thiosulfinate. As used herein, "substantially free" refers to compositions containing less than 5% by weight, preferably less than 1% by weight, more preferably less than 0.5% by weight, of diallyl thiosulfinate, for example when referring to a concentrated composition having 50% or more PTSO. Dilute working solutions will contain significantly less diallyl thiosulfinate.

[0076] The composition of the present disclosure is also preferably substantially free of diallyl disulfide.As used herein, "substantially free" refers to a composition that contains less than 5% by weight of diallyl disulfide, preferably less than 1% by weight, more preferably less than 0.5% by weight, when referring to a concentrated composition, such as a concentrated composition having 50% or more PTSO.Diluted working solutions will contain significantly less diallyl disulfide.

[0077] The composition may include any suitable "agriculturally acceptable carrier, excipient and / or solvent." Such carriers and solvents are known to those skilled in the art and are not unacceptably damaging to the plant or its environment and / or unsafe for the user or other people to whom they may be exposed. For example, the agriculturally acceptable carrier may be a solid carrier, a gel carrier, a liquid carrier, a suspension, or an emulsion. A non-limiting example of a solvent is water.

[0078] In a preferred embodiment, the composition further comprises an emulsifier. Suitable emulsifiers include propylene glycol and polyethylene glycol glyceryl ricinoleate. Yuca extract and Tween are also suitable emulsifiers. In some embodiments, the composition comprises 40-70% emulsifier, preferably 55-60% emulsifier, particularly where the emulsifier is a combination of propylene glycol and polyethylene glycol glyceryl ricinoleate. In some embodiments, the composition comprises 70-92% emulsifier, preferably about 90% emulsifier, particularly where the emulsifier is Yuca extract. In some embodiments, the composition comprises 25-70% emulsifier, preferably 25-35% emulsifier, particularly where the emulsifier is Tween. Preferably, the composition comprises 1-30%, preferably 3-10%, of a compound as disclosed herein (e.g., PTSO). The compositions may further include vitamins and minerals, such as vitamin H and vitamins B1, B2, B3, B5, B6 and B12. Such compositions can be diluted into working solutions for application to plants, as further disclosed herein.

[0079] In some embodiments, the composition further comprises a fertilizer, an insecticide, a wetting agent, an antimicrobial compound, a fungicide, a chelating compound, an aromatic compound, and / or an additional biostimulant.

[0080] Those skilled in the art will also understand that fertilizers, insecticides, wetting agents, antimicrobial compounds, fungicides, chelating compounds, aromatic compounds, and / or additional biostimulants (particularly amino acid-based biostimulants as described herein) can also be provided in separate compositions. For example, the present disclosure further contemplates a method comprising providing the plant with a compound or composition of the present invention disclosed herein, and providing the plant with one or more additional agents selected from fertilizers, insecticides, wetting agents, antimicrobial compounds, fungicides, chelating compounds, aromatic compounds, and / or additional biostimulants. Preferably, the additional agents are provided at about the same time as the compound of the present invention. The additional agents may also be provided before or after the compound of the present invention, for example, hours or days before or after the compound of the present invention. Preferably, the compound of the present invention and the additional agents are provided within 7 days of each other. The additional agents are preferably provided multiple times during the crop cycle. For example, the additional agents may be applied every 4 to 14 days. The additional agents may be applied at least four times, preferably at least six times, during the crop cycle. In particular, the composition comprising formula I and the additional agent are both provided during the same crop cycle.

[0081] In some embodiments, the methods and uses disclosed herein further comprise applying an amino acid-based biostimulant to the plant. As referred to herein, an "amino acid-based biostimulant" comprises at least 10% (w / w), preferably at least 15% (w / w), of amino acids. In some embodiments, the biostimulant comprises at least 50%, preferably at least 55% (w / w), of amino acids. The amino acids may be free amino acids (i.e., amino acids in free form) or may be bound to other amino acids (e.g., via peptide bonds). In some embodiments, the amino acid biostimulant comprises 8-15% (w / w) of free amino acids and 45-55% of attached amino acids. Such biostimulants are generally diluted 1:10 to 1:1,000 before use.

[0082] Amino acid-based biostimulants and their methods of use are known to those skilled in the art and include Metalosate Calcium and Metalosate Fe (Albion Minerals, Layton, UT, USA); Agrocean B (Agrimer, Plouguerneau, France); Tecamin Brix, Tecamin Max, Tecnokel Amino Mix, and Terra-Sorb Foliar (Agritecno Fertilizantes, Valencia, Spain); Amino Quelant Ca (Bioiberica, Barcelona, ​​Spain); Bosfoliar Activ (COMPO EXPERT, Munster, Germany); NaturalCrop SL (NaturalCrop Poland Sp.z oo, Warszaw, Poland); and Delfan Plus (Tradecorp, Madrid, Spain). See Table 2 in Molecules. 2018 Feb;23(2):470 for a description of the composition of the amino acid biostimulants AminoPrim and AminoHort (containing 15% and 20% amino acids, respectively, and 0.27% and 2.1% trace elements, respectively).

[0083] TerraSorb 商標 The Terra Sorb Complex is another suitable amino acid-based biostimulant that contains 20% (w / w) of the following free amino acids: ASP, SER, GLU, GLY, HIS, ARG, THR, ALA, PRO, CIS, TYR, VAL, MET, LYS, ILE, LEU, PHE, and TRP. The Terra Sorb Complex also contains 5.5% nitrogen (of which 5% is organic nitrogen), as well as B (1.5%), Mg (0.8%), Fe (1%), Zn (0.1%), Mn (0.1%), Mo (0.001%), and 25% organic matter.

[0084] The preferred amino acid-based biostimulant is Isabion 商標 Isabion 商標is a mixture of water, ash, free amino acids, and short and long chain peptides.

[0085] Preferably, Isabion 商標 is a mixture of 33.5% (w / w) water, 4% (w / w) ash, and 62.50% (w / w) organic matter. In particular, the mixture contains 10.3% (w / w) free amino acids and 47.96% (w / w) attached amino acids. The free amino acids are 3.80% (w / w) glycine, 1.45% (w / w) proline, 1.87% (w / w) alanine, 0.27% (w / w) glutamic acid, 0.85% (w / w) hydroxyproline, 0.35% (w / w) aspartic acid, 0.20% (w / w) leucine, 0.35% (w / w) lysine, 0.09% (w / w) sucrose ... (w / w) valine, 0.33% (w / w) tyrosine, 0.16% (w / w) phenylalanine, 0.07% (w / w) isoleucine, 0.12% (w / w) arginine, 0.08% (w / w) threonine, 0.08% (w / w) methionine, 0.10% (w / w) histidine and 0.13% (w / w) serine.

[0086] The attached amino acids were 8.65% (w / w) glycine, 8.78% (w / w) proline, 5.16% (w / w) alanine, 6.33% (w / w) glutamic acid, 5.35% (w / w) hydroxyproline, 2.71% (w / w) aspartic acid, 1.90% (w / w) leucine, 1.68% (w / w) lysine, and 1.67% (w / w). (w / w) valine, 1.14% (w / w) tyrosine, 1.18% (w / w) phenylalanine, 0.87% (w / w) isoleucine, 0.80% (w / w) arginine, 0.66% (w / w) threonine, 0.57% (w / w) methionine, 0.37% (w / w) histidine and 0.14% (w / w) serine.

[0087] Furthermore, the attached amino acid can be a short peptide and / or a long peptide. The molecular weight of the short peptide is generally about 1160 Da to about 3500 Da. The molecular weight of the long peptide is generally about 3600 Da to about 8500 Da.

[0088] Isabion 商標 is usually diluted with water and applied as a foliar spray or in irrigation water. 商標 is usually diluted to 200-300ml / 100L of water.

[0089] Isabion 商標 The recommended dose is 200ml / hl to 300ml / hl for foliar sprays and 2L / ha to 3L / ha for irrigation systems. In case of frost or crop damage, the dose can be increased to 400ml / hl for foliar sprays and 4L / ha for irrigation systems.

[0090] [Table A]

[0091] In an exemplary embodiment, providing a plant with a compound according to Formula I or a composition comprising a compound according to Formula I, and administering an amino acid-based biostimulant, such as Isabion, to the plant. 商標 and providing said plant with

[0092] In some embodiments, the compound according to Formula I and the amino acid-based biostimulant are provided in a single composition. Thus, the present disclosure provides a composition comprising a compound according to Formula I and an amino acid-based biostimulant.

[0093] In some embodiments, the composition comprises a compound according to Formula I, glycine and proline. In some embodiments, the composition comprises a compound according to Formula I and a free amino acid selected from the group consisting of glycine and proline. Preferably, the composition comprises a compound according to Formula I, a free amino acid selected from the group consisting of glycine and proline, and a peptide.

[0094] Preferably, the composition further comprises a free amino acid selected from the group consisting of alanine, glutamic acid and hydroxyproline; more preferably, the composition even further comprises a free amino acid selected from the group consisting of aspartic acid, leucine, lysine, valine, tyrosine, phenylalanine, isoleucine, arginine, threonine, methionine, histidine and serine.

[0095] In some embodiments, compositions are provided that include a compound according to Formula I; the free amino acids alanine, glutamic acid, hydroxyproline, aspartic acid, leucine, lysine, valine, tyrosine, phenyl-alanine, isoleucine, arginine, threonine, methionine, histidine, serine, glycine, and proline; and a peptide.

[0096] In a preferred embodiment, the composition comprising formula I described herein and the amino acid-based biostimulant described herein are provided as separate compositions. Preferably, the composition comprising a compound according to formula I is provided at least 24 hours (or 1-3 days) before or 24 hours (or 1-3 days) after the amino acid-based biostimulant is provided to the plant. Also provided is a kit of parts comprising i) a composition comprising formula I described herein and ii) an amino acid-based biostimulant described herein.

[0097] In a preferred embodiment, the composition comprising formula I further comprises cadmium (Cd) in an amount of 1.5 mg / kg or less of dry matter. In a preferred embodiment, the composition comprises hexavalent chromium (Cr VI) in an amount of 2 mg / kg or less of dry matter. In a preferred embodiment, the composition comprises lead (Pb) in an amount of 120 mg / kg or less of dry matter. In a preferred embodiment, the composition comprises mercury (Hg) in an amount of 1 mg / kg or less of dry matter. In a preferred embodiment, the composition comprises nickel (Ni) in an amount of 50 mg / kg or less of dry matter. In a preferred embodiment, the composition comprises inorganic arsenic (As) in an amount of 40 mg / kg or less of dry matter. In a preferred embodiment, the composition comprises inorganic zinc (Zn) in an amount of 1500 mg / kg or less of dry matter.

[0098] Dimethylthiosulfonate, Diphenylthiosulfonate

[0099] The present disclosure provides organosulfur compounds and compositions comprising the compounds disclosed herein for use as plant biostimulants.The compounds and compositions can be used in methods for treating plants.Such methods can be useful for increasing the growth rate and / or development of plants, increasing the yield of plants, or increasing the harvest of plants.

[0100] In some embodiments, a method is provided that includes providing a compound or composition disclosed herein to a plant. The plant can be provided with the compound or composition by any means known in the art, such as topical nutrition through roots, flowers, leaves and stems, watering nutrition, spraying or wetting solution, coating, piling, aerial solution, epidermis, intravascular, etc. The preferred route is uptake through roots, spraying, aerial administration or topical administration. The preferred route of administration is through the roots using irrigation (e.g., drip / drip irrigation). Without wishing to be bound by theory, providing the compound directly to the roots may provide improved efficacy than, for example, spraying the leaves. Table 17 below compares the results between spraying and drip irrigation. Spraying the plant is also effective, especially when the composition can penetrate into the ground and reach the roots.

[0101] In some embodiments, the compounds of the present invention are provided multiple times during the crop cycle, which as used herein refers to the time from germination to harvest of the crop.

[0102] For example, the compound may be applied at least 4 times, preferably at least 6 times, during the crop cycle. Preferably, the compound is applied every 4-27 days, particularly every 4-14 days. In an exemplary embodiment, the compound is provided every 1-2 weeks. In an exemplary embodiment, the compound is provided every 4-27 days, preferably every 4-14 days, and at least 4 times, preferably at least 6 times, during the crop cycle. The first application of the compound is preferably immediately after germination. The effect of the compound continues for several weeks after the last application, but it is recommended to apply the compound at least 7 days before the planned harvest date.

[0103] The compound or composition can be applied to plants (including cuttings, emerged seedlings, and established vegetation, including roots and above-ground parts such as leaves, stems, flowers, fruits, branches, limbs, roots, etc.), to plant seeds (e.g., before germination), or to surrounding soil, particularly to the root zone of plants.As used herein, the term "rhizosphere" refers to the area of ​​soil adjacent to the root of a living plant.The width of the root zone is generally within 100 mm from the surface of the root.

[0104] The compound or the composition can be administered in a single dose or administered as multiple doses.For example, the composition can be provided daily, weekly, monthly or yearly.In exemplary embodiments, the composition can be provided once a day, for a week or until the biostimulant is effective.

[0105] Watering of plants is performed as a watering turn, and the amount of water supplied is selected so that the plant will excrete a portion of the water supplied as wastewater. This means that the organosulfur composition will be washed away relatively quickly. To achieve a uniform concentration in the root support, the organosulfur composition can be administered as granules that release the composition at a desired rate. In this manner, a constant concentration is provided over an extended period of time.

[0106] In some embodiments, the compositions disclosed herein are provided as spray solution.When the composition is sprayed on plants, the solution can be deposited on leaves as droplets with small surface-to-volume ratios that evaporate, which can cause the composition to remain on leaves as residue.This effect can be reduced by including a wetting agent in the composition.

[0107] The amount of the organosulfur compound to be applied depends on a variety of factors, including the method of application, the time of application, the rate of decomposition of the particular compound used, the duration of treatment, the insecticide treatment, the compounds and / or materials used in combination, the age, weight, general health, and previous treatments, as well as factors well known in the agricultural arts. A horticulturist, plant grower, or farmer having ordinary skill in the art can readily determine the effective amount of the composition required.

[0108] It is clear to those skilled in the art that lower concentrations / amounts of organosulfur compounds can be administered to slow-growing plants, such as cacti and succulents. It is also clear to those skilled in the art that the concentration / amount in water and the frequency of application depend on the plant species, subspecies, cultivars, hybrids, and variants. Moreover, it is clear to those skilled in the art that the dosage that the plant can tolerate depends on the growth stage and size of the plant. Moreover, it is clear to those skilled in the art that the concentration / amount in water and the frequency of application depend on the growth conditions, such as light, temperature, evaporation, nutrient concentration and pH in the root support, air movement, and application of other insecticides. Moreover, it is clear to those skilled in the art that the concentration / amount in water and the frequency of application depend on the moment it is applied, day, night, season, and weather.

[0109] In an exemplary embodiment, the organosulfur compound, preferably PTSO, is provided in an amount of from 0.01 kg / ha to 100 kg / ha.

[0110] Preferably, for use in orchids, preferably Phalaenopsis, the organosulfur compound, preferably PTSO, is provided in an amount of from 10 kg / ha to 100 kg / ha, more preferably from 20 kg / ha to 60 kg / ha. Preferably, for use in Chrysanthemum, the organosulfur compound, preferably PTSO, is provided in an amount of from 0.01 kg / ha to 1 kg / ha, more preferably from 0.01 kg / ha to 0.25 kg / ha.

[0111] In some embodiments, a compound of formula I, or a composition comprising the compound, is applied directly to the plant, to the seeds of the plant, or to the soil of the plant or seeds.

[0112] As demonstrated in the Examples, the compounds disclosed herein have beneficial effects on plants, particularly on plant quality characteristics, and are therefore useful as plant / soil additives, fertilizers, and biostimulants.

[0113] In a preferred embodiment, the term "biostimulant" refers to a product, in particular a compound or composition, that stimulates the nutritional processes of a plant, independently of the nutrient content of the product, with the aim of improving one or more of the following properties of the plant or the rhizosphere of the plant: a) nutrient utilization efficiency, b) biological stress resistance, c) quality traits (also known as quality characteristics), and d) availability of trapped nutrients in the soil or rhizosphere.

[0114] In a preferred embodiment, the use of the compounds described herein as biostimulants comprises a) nutrient use efficiency, b) resistance to abiotic stress, c) quality traits, and d) availability of trapped nutrients held in the soil or in the rhizosphere. The present invention relates to improving one or more of the following:

[0115] In a preferred embodiment, use as a biostimulant does not include use as an antibacterial and / or antifungal agent.

[0116] Preferably, the organosulfur compounds disclosed herein are Increase nutrient use efficiency, and / or Increase plant tolerance to abiotic stresses, and / or improve the quality characteristics of plants, and / or Increases the availability to plants of nutrients held in the soil or in the rhizosphere.

[0117] More preferably, the organosulfur compounds disclosed herein are Increase nutrient use efficiency, and / or Increase plant tolerance to abiotic stresses, and / or Improve plant quality characteristics.

[0118] Nutrient Use Efficiency

[0119] As used herein, nutrient utilization efficiency is used according to the usual definition in the art. Typically, nutrient utilization efficiency is defined as yield (biomass) per unit input (fertilizer, nutrient content) (see, for example, Nutrient Use Efficiency in Plants, Concepts and Approaches, Editors: Hawkesford, Malcolm J., Kopriva, Stanislav, De Kok, Luit J. (Eds.) ISBN 978-3-319-10635-9). Preferably, the nutrient is nitrogen (N) and / or phosphorus (P).

[0120] Those skilled in the art are familiar with the methods for determining plant nutrient utilization efficiency, and therefore can also fully establish whether biostimulants enhance said nutrient utilization efficiency.For example, in field studies, nutrient utilization efficiency can be calculated based on the difference in crop yield and / or nutrient uptake between a field treated with biostimulants and an untreated control field, or can be calculated by estimating the crop and soil recovery of applied nutrients using isotope-labeled nutrients (see, for example, A. Dobermann, Nutrient use efficiency - measurement and management: Fertilizer Best Management Practices, 2007, ISBN: 2-9523139-2-X).

[0121] Abiotic stress

[0122] As used herein, abiotic stress is defined as the negative effect of non-biotic factors on living organisms in a particular environment. Thus, improving the resistance of a plant to abiotic stress indicates that the plant is more able to withstand abiotic stress factors that usually have a negative effect on plants, such as a decrease in plant growth rate. Abiotic stress factors include, but are not limited to, drought, excess water (especially too much rain or too much humidity), too much or too little direct sunlight, strong winds, suboptimal soil structure, salinity (especially excessive salinity), too low or too high temperature, strong and / or sudden fluctuations in temperature, and other environmental extremes. Meanwhile, plant pathogens or pests are called biotic stress.

[0123] Adverse effects on plants resulting from abiotic stress include, but are not limited to, reduced plant growth rate, reduced maximum plant height, reduced root formation, poor leaf development (e.g., smaller leaves, different leaf color), increased susceptibility to insect or pathogen damage, and smaller flowers or flower stems.

[0124] The average improvement in the resistance of plants to abiotic stress as a result of treating the plants with a biostimulant can be measured by comparing a field with plants treated with a biostimulant to an untreated control, where the treated plants and the control plants are subjected to substantially the same abiotic stress factor, e.g., when the treated plants and the control plants are grown in the same field or adjacent fields.

[0125] In a preferred embodiment, the abiotic stress is drought, excess water, direct sunlight, heat, and / or cold in Phalaenopsis plants, which results in slowed growth, reduced root formation, pale or red leaves, increased susceptibility to insect or pathogen attack, and / or reduced flowers or flower stems.

[0126] In a preferred embodiment, the abiotic stress is related to too little nutrient uptake or too little water in Chrysanthemum plants due to insufficient and slow root formation due to lack of sufficient roots, too high temperature, and / or air movement.

[0127] Quality traits

[0128] The definition of a quality trait (also known as a quality characteristic) may depend on the plant genus or species. Typically, a person skilled in the art, e.g. a farmer or grower, knows which plants are associated with which quality traits.

[0129] Some examples of crops / ornamental plants and their quality attributes are given below. Other quality attributes are given in Table 31 below.

[0130] [Table B]

[0131] The term "improvement in plant quality" refers to a qualitative or quantitative improvement in a trait when compared to the same trait in a control plant grown under the same conditions in the absence of application of the compounds disclosed herein. Such traits include, but are not limited to, improved visual appearance of the plant, improved quality of the harvested product, such as seeds, fruits, leaves, vegetables; improved visual appearance of the harvested product, improved nutritional content, increased shelf life, etc.

[0132] In some embodiments, the plant quality is plant vigor.Improved plant vigor includes, for example, improved plant vitality of the plant; improved plant stand; improved heading; and / or more developed root system; enhanced nodules; larger leaf blades; improved leaf color, increased plant size; increased plant weight; increased plant height; increased yield when grown in poor soil or adverse climate; earlier flowering / fruiting / germination / grain maturity; faster and more uniform maturation, etc.

[0133] In some embodiments, the quality of the plant refers to the quantity of the plant, such as the yield of the plant (e.g., grains, nuts, fruits, vegetables, seeds, etc.). Improved plant yield includes, for example, an increase in biomass production and an increase in the harvestability of the plant.

[0134] In some embodiments, improved quality characteristics refer to one or more characteristics selected from improved visual appearance of the plant, improved quality of the harvested product, improved nutritional content, increased shelf life, improved plant vigor, and improved plant yield.

[0135] Availability of trapped nutrients held in the soil or in the rhizosphere

[0136] As those skilled in the art will recognize, some of the nutrients may be trapped in the soil or rhizosphere, making a certain amount of nutrients unavailable to plants. Trapped nutrients include, but are not limited to, nutrients that have low mobility in the soil or rhizosphere and / or are poorly soluble in water. Low mobility may be, for example, the result of the nutrient interacting with other soil components, such as clay-sized particles or organic matter associated with minerals. The physicochemical interactions of nutrients with other soil components may limit the availability of nutrients to plants (Jilling et al.Biogeochemistry (2018) 139:p.103-122).

[0137] A variety of methods are available to measure the availability of trapped nutrients in soils and the rhizosphere (see, e.g., Brinkley and Vitousek, Soil nutrient availability, in: Plant Physiological Ecology: Field Methods and Instrumentation (ed. by Pearcy, Ehleringer, Mooney, and Rundel), 2000, ISBN: 13:978-0-412-40730-7).

[0138] On top of that, To increase the growth rate and / or development of plants, To increase the yield of plants, To increase the yield of plants As will be appreciated by one of ordinary skill in the art, an increase in the growth rate of a plant, for example, refers to an increase in the growth rate of a plant treated with a compound disclosed herein compared to the growth rate of a plant grown under similar conditions without treatment.

[0139] As used herein, the term "plant" includes crop plants, ornamental plants, trees, grasses, annuals, perennials, or other commonly cultivated members of the Plant Kingdom. As used herein, the term "crop plants" includes plant species that have commercial value and are planted and cultivated for commercial use. Thus, crop plants include flowering and non-flowering plants, perennials and annuals, trees, shrubs, vegetable plants, fruit trees, turf, and ground cover plants.

[0140] Suitable plants include Cymbidium, Oncidium, Miltonia, Paphiopedilum, Cypripedium, Calanthe and hybrids of the orchid genus, Bromelia, Begonia, Impatiens, Azalea, ferns; horticultural crops such as sweet pepper, tomato, eggplant, cucumber, zucchini, etc.; arable crops such as wheat, potato, beet, chore, Luzerne, etc.; arable horticultural crops such as endive, cauliflower, Brussels sprouts, etc. sprouts, lettuce, broccoli, chicory, peas, beans, red cabbage, kale, etc.; garden trees, such as azalea, magnolia, forsythia, peony, hollyhock, laburnum, palm, wisteria, etc.; fruit trees and shrubs, such as apple trees, pear trees, cherry trees, prune trees, gooseberry, black currant, blueberry trees, cranberry trees, etc.; tropical fruits, such as bananas, papayas, cassava, pineapple, avocado, mango, etc.

[0141] Preferably, the plant belongs to the clade Embryophyta. Preferably, the plant is a vascular plant. In an exemplary embodiment, the plant is a crop plant, such as Lactuca sativa (lettuce).

[0142] In another preferred embodiment, the plant belongs to the clade Angiospermae. Preferably, the plant belongs to a family selected from the group consisting of nightshades (Solanaceae), cucumbers (Cucurbitaceae), roses (Rosaceae), orchids (Orchidaceae), lilies (Liliaceae), composites (Asteraceae or Composite), carnations (Caryophyllaceae), crucifers (Brassicaceae or Cruciferae), grasses (Poaceae), and umbellifers (Apiaceae or Umbelliferae).

[0143] In a preferred embodiment, the plant belongs to the family of nightshades (Solanaceae). Preferably, the plant belongs to the genus Solanum or Capsicum. Preferably, the plant belongs to the genus Solanum and is selected from the group consisting of tomato (S. lycopersicum), potato (S. tuberosum), eggplant (S. melongena) and pepino (S. muricatum). In another preferred embodiment, the plant belongs to the genus Capsicum and belongs to the pepper species (C. annuum), in particular sweet pepper, chili pepper and jalapeno.

[0144] In a preferred embodiment, the plant belongs to the Cucurbitaceae family. Preferably, the plant belongs to the genus Cucumis or Cucurbita. Preferably, the plant belongs to the genus Cucumis and is selected from the group consisting of cucumber (C. sativus), sugar melon and gherkin (C. anguria). In another preferred embodiment, the plant belongs to the genus Cucurbita and is selected from the group consisting of C. pepo (especially zucchini) and pumpkins (C. argyrosperma, C. digitate, C. maxima and C. moschata).

[0145] In a preferred embodiment, the plant belongs to the family Rosa ceae. Preferably, the plant belongs to the family Fragaria, Pyrus, Malus, Rosa, and raspberry (particularly the subfamily Rubus and blackberries).

[0146] In a preferred embodiment, the plant belongs to the orchid family (Orchidaceae). In an exemplary embodiment, the plant is an ornamental plant, such as an orchid, in particular from the Phalaenopsis genus, or other flowering plants, such as those from the Cymbidium genus.

[0147] In a preferred embodiment, the plant belongs to the lily family (Liliaceae). Preferably, the plant belongs to the genus Tulipa or Lilies (Lillium).

[0148] In a preferred embodiment, the plant belongs to the composite family (Asteraceae or Compositae). Preferably, the plant belongs to the genus Chrysanthemum, Asterea or Lactuca.

[0149] In a preferred embodiment, the plant belongs to the carnation family (Caryophyllaceae).

[0150] In a preferred embodiment, the plant belongs to the crucifers family (Brassica ceae or Cruciferae). Preferably, the plant belongs to the genus Brassica.

[0151] In a preferred embodiment, the plant belongs to the grass family (Poaceae). Preferably, the plant belongs to the genus Triticum, Oryza or Zea.

[0152] In preferred embodiments, the plant belongs to the umbellifers family (Apiaceae or Umbelliferae). Preferably, the plant is selected from the group consisting of carrots (Caucus carota), parsnip (Pastinaca sativa), anise (Pimpinella anisum), coriander (Coriandrum sativum), and cumin (Cuminum cyminum). In some embodiments, the plant is not cumin.

[0153] Preferably, the plant is selected from the group consisting of the genus Phalaenopsis, Cymbidium, Chrysanthenum, Rosa, Fabaceae (preferably Phaseolus, more preferably Phaseolus vulgaris), Brassica (preferably Brassica oleracea and Brassica rapa), Cucurbita (preferably Cucurbita pepo giromontiina, Cucumus melo, and Cucumus sativus), Solanaceae (preferably Solanum or Capsicum, more preferably Solanum lycopersicum or Capsicum annuum), Vitis (preferably Vitis vinifera), Vaccinia (preferably Vaccinia corymbosum or Vaccinium cyanococcusi), and Lactuca (preferably Lactuca sativa).

[0154] As used herein, the word "comprise" and its conjugations are used in an open-ended sense, meaning that the items following the word are included, but items not specifically mentioned are not excluded. In addition, the verb "consisting of" may be replaced by "consisting essentially of," meaning that the compound or auxiliary compound defined herein may contain one or more additional components than those specifically identified, where the one or more additional components do not alter the inherent properties of the present invention.

[0155] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0156] The words "approximately" or "about" when used in connection with a numerical value (such as approximately 10, about 10) preferably mean that the value may be greater than or equal to 1% of the given value of 10 or less than 1% of the value.

[0157] The present invention will be further described in the following examples, which are not intended to limit the scope of the invention but are merely intended to clarify the invention.

[0158] Working Example

[0159] This experiment demonstrated for the first time the effectiveness of PTSO / PTS as a biostimulant. Although the growing conditions (watering, fertilization, light, temperature, air movement, air humidity) were optimal for ornamental potted Phalaenopsis hybrid "Pure silk", the administration of various amounts of PTSO / PTS resulted in higher growth rates and improved plant characteristics, as demonstrated in Example 2 below. During periods of high temperature and light in the greenhouse, the treated plants continued to grow at a high rate, while the growth of untreated plants slowed down. Apparently, the plants treated with PTSO were better able to resist periods of increased abiotic stress.

[0160] Example 3 below shows that treating Chrysanthemum cuttings with various amounts of PTSO / PTS resulted in increased growth and better root development. In this experimental design, treated cuttings were compared to untreated cuttings under the same conditions for optimal rooting and pre-growth of the cuttings. In this example, treatment with PTSO / PTS demonstrated increased rooting and growth compared to untreated cuttings under optimal conditions. Moreover, these experiments also showed the additional effect of more efficient nutrient uptake, indicating that the PTSO / PTS composition used herein is a biostimulant.

[0161] Example 1. Separation of PTSO and PTS

[0162] range:

[0163] In many of the examples described herein, an aqueous extract of PTSO is used, referred to herein as PTSO extract (PE), which contains at least 56% PTSO (wt %) and no more than 14% PTS (wt %).

[0164] Study design: The elution order of the compounds according to their polarity is deduced from high-performance liquid chromatography (HPLC). The conditions for separation are determined by thin-layer chromatography (TLC) using heptane and ethyl acetate as mobile phase. A flash column of 110 cm length and 25 cm diameter is loaded with 15 kg of silica (ACROS Organics) in heptane. 商標 The column was packed with 40-60 μm-particle size cellulose (from Sigma-Aldrich) and allowed to stabilize overnight. Purification was performed by gradually increasing the polarity from 0% to 40%. 1 The fractions of interest, namely PTS (= peak 6) and PTSO (= peak 7), both identified by H-NMR, began to elute after 20 liters of mobile phase and were collected in fraction sizes of 2 liters. The fractions were separated according to the TLC identification of each compound, and the corresponding fractions for each compound (PTS or PTSO) were combined. The solvent was evaporated by rotary evaporation to obtain a purified fraction of PTS (32435-2-A) and two purified fractions of PTSO (MHA32435-2-B, MHA32435-2-C) as outlined in Table 1.

[0165] The required purity of >98% was achieved only in fraction MHA32435-2-B (=PTSO), whereas fractions MHA32435-2-A (PTS) and MHA32435-2-C (=second fraction of PTSO) showed purities below 98% and required further purification by a second flash chromatography run to achieve the desired purity.

[0166] The repurification of PTS (MHA32435-2-A) was carried out by loading 2 kg of silica (particle size 40-60 μm, ACROS Organics) into a flash column (length 50 cm, diameter 20 cm). 商標 ), with ethyl acetate and heptane as the mobile phase and increasing the polarity from 0% to 20%. Fractions of interest were collected in fraction sizes of 250 ml, identified by TLC, and combined. The solvent was evaporated by rotary evaporation to obtain PTS (EWR32514-01-1) with a purity of >98%.

[0167] Repurification of the PTSO impurity fraction (MHA32435-2-B) was performed on the same flash column using the same mobile phase as for MHA32435-2-A, except that 3 kg of silica (for column packing) was used, the polarity was increased from 0% to 30%, and the fraction size was 100 ml. The repurification resulted in two fractions of PTSO (EWR32514-02-1 and EWR32514-02-02) with a purity of over 98%, as outlined in Table 1.

[0168] [Table 1]

[0169] This example demonstrates that PTSO and PTS were purified to high purity.

[0170] In the case of Examples 2-5 described below, the PTSO extract (PE) contains 56% PTSO and approximately 30% polyethylene glycol glyceryl ricinoleate.

[0171] Example 2. Treatment of ornamental plants

[0172] Scope: In this example, the biostimulatory effect of PTSO extract was demonstrated. For this purpose, Phalaenopsis "Pure silk" was selected. Plant growth and development involves three stages: (1) vegetative growth, (2) flower induction, and (3) flower stalk development.

[0173] The plants were evaluated at the end of the flowering induction period following the treatments described below.At the time of sale, the plants were evaluated for flowering.

[0174] Plant Cultivation Procedure

[0175] Phalaenopsis "Pure silk" plants were produced from meristems and pre-grown. Plants delivered to Phalaenopsis growers were treated according to the following procedure: they were potted in clear 12 cm pots in medium-sized bark (supplier: Bas van Buuren BV, De Lier), cultivated at 29°C for 30 weeks, and watered and fertilised (100 litres / m2) every 5 days. 2 ) was showered (EC 1.1).

[0176] If necessary, the plants were exposed to artificial light (7 am to 6 pm). For flowering induction, the plants were chilled and grown at 19 °C for 2 months. After that, the plants were grown at 20-25 °C for 3-4 months and exposed to artificial light optionally if necessary. In this way, flowering plants of appropriate size according to market demand were obtained.

[0177] In July 2020, a young Phalaenopsis orchid was potted in a bark. July was a hot and sunny month, and despite the grower's use of a shade screen, the plant exhibited dark green / reddish droopy thin leaves and little growth. This indicated that the plant was experiencing abiotic stress. Root number and root growth were observed by visual inspection through a transparent pot. Considerable variation in root number and root growth was observed within the group of plants.

[0178] The following experimental design was set up to determine the sensitivity of plants to various dilutions and treatment times with PTSO extract. 2 An area (containing about 100 plants) was treated with diluted solutions of PTSO / PTS as follows, and then watered (10 L / m 2 ) was manually showered. Manual watering with diluted PTSO extract was done directly after regular showering for 5 days. In this way, the moisture content of root supports of control and treated plants was almost the same.

[0179] Treatment 1:: Two treatments with 1,000-fold diluted feeding solution of PTSO extract (EC 1.1), given every other watering (10-day interval); Treatment 2: two treatments with 2,500-fold diluted feeding solution of PTSO extract (EC 1.1), given every other watering (10-day interval); Treatment 3: eight treatments with 1,000-fold diluted feeding solution of PTSO extract (EC 1.1), given every other watering (10-day interval); Treatment 4: eight treatments with 2,500-fold diluted feeding solution of PTSO extract (EC 1.1), given every other watering (10-day interval); After treatment, the treated plants underwent the same cultivation procedures as the control, i.e., untreated, plants.

[0180] After 36 weeks, the lines were scored for the following characteristics:

[0181] Differences between plants were noted in root mass, root stump growth, leaf progression, leaf number and length, plant and bark quality (assessed by color and water retention, i.e. estimated weight). Evaluation was performed as follows: From the control plants, the largest and smallest plants were selected. The largest was rated 5 and the smallest was rated 1.

[0182] For rating root formation the same scoring procedure was followed: plants with the most roots were rated 5 and plants with the least roots were rated 1.

[0183] The length of the largest fully developed leaf was measured with a tape measure.

[0184] "6-7 leaves" means that 6 leaves are already fully developed and the 7th leaf is still growing.

[0185] "Leaf thickness" was scored according to the resistance of the leaf when arched by hand.

[0186] "Bark quality" was scored by judging the degree of composition: the wetter and muddier the bark, the lower the score.

[0187] "Progressive growth" is an estimate of how much larger each leaf on a plant is than the previous leaf, specifically the last fully developed leaf compared to the penultimate developed leaf.

[0188] "Plant size" was scored as follows: the larger the plant, the higher the score.

[0189] In the case of "plant characteristics", the focus was on the size of the plant.

[0190] Control plants were rated 1-5 and compared to the treated plants. If any one of the treated plants performed better on the evaluation criteria, they could be scored higher than 5. The plants with the worst "plant traits" were rated "1" and the best plants were rated "5".

[0191] Results and Discussion

[0192] The average measurements are summarized in Table 2 below and are based on the raw data. Data was collected 36 weeks after treatment, the plants had just left the refrigerated portion of the greenhouse for 2 weeks for flowering induction.

[0193] [Table 2]

[0194] Plants treated twice with 1,000 and 2,500 dilutions of PTSO extract showed significantly less root growth compared to the control. Although the roots of these treated plants were less developed than those of the control, the roots of the treated plants allowed them to grow faster.

[0195] Similar, but more striking, observations were made for the plants treated 8 times with both dilutions. The size of the plants was the largest among all treatments. In addition, the roots showed a fairly good development. Moreover, not only were there more roots, but the root growth tips were larger and thicker. It was concluded that the treatment with 8 applications of the 1,000-fold dilution had the greatest effect on the Phalaenopsis plants.

[0196] The quality of the root support bark was also scored, as shown in Table 1. Bark is gradually decomposed by microbial activity during plant cultivation. As the bark decomposes, it turns into moist anaerobic material, which can lead to root death. Table 1 shows that the bark quality is slightly lower for the treated plants (2.72-3) compared to the control (3.18), but these differences are so small that no adverse effects on the treated plants are expected.

[0197] conclusion

[0198] A very significant effect of the PTSO extract on the properties of the scored strains was demonstrated. Growth stimulation was observed compared to the control strain. Since plant growth hormones are not contained in the PTSO extract, this composition falls under the category of biostimulants. During the experiment, which resulted in abiotic stress (there were periods of drought, heat, and very strong light), it was clear that the strains could not grow at their maximum growth rate. In the experiment, it was shown that the PTSO extract has the ability to compensate for abiotic stress.

[0199] Although the fertilization conditions were exactly the same for all plants, the plants treated with PTSO extract had improved plant characteristics and more plant biomass was formed, indicating that the nutrient utilization efficiency for the plants treated with PTSO extract was higher.

[0200] Another interesting observation was that the plant had a very pronounced shine on the leaves. This was attributed to the formation of a thicker wax layer on the leaves, which is thought to indicate better health of the plant. Without wishing to be bound by theory, it is believed that a thicker layer of wax typically reduces water evaporation from the upper leaf surface, which contributed to the reduction in abiotic stress.

[0201] Post-treatment observations of Phalaenopsis plants treated twice with PTSO extract showed that it was sufficient to induce long-term effects over a cultivation period of more than six months, which was quite surprising.

[0202] Flowering of the plant was not affected by the treatments and the plant has become an excellent commercial product. No wilting, flower abnormalities, or abnormalities in stem shape or size were observed.

[0203] In this example, it is clearly shown that the PTSO extract is a potent biostimulant.

[0204] Example 2b. Treatment of Phalaenopsis strains

[0205] Scope: In this example, survival time after repotting and production of flower stalks during the cultivation period were demonstrated. For this purpose, various Phalaenopsis hybrids were selected. Plant growth and development included three stages: (1) vegetative growth, (2) flower induction, and (3) flower stalk development. The same grow-up regime was followed as described in Example 2. After the treatments described below, the failure rate after repotting and the number of flower stalks at the end of the cultivation cycle were evaluated.

[0206] Scope: During Phalaenopsis cultivation, growers face significant losses after transplantation. These can reach up to 10%, depending on the size, condition and genetic background of the hybrid. In this example, it was demonstrated that PTSO extract significantly reduces the failure of (young) Phalaenopsis after transplantation and increases the number of flower stalks before the sale of the plants. For this purpose, different hybrids of Phalaenopsis were grown from meristems and pre-grown in 70-hole trays in Floricultura until they reached a diameter of 5 cm. The plants were then handed over to growers and transplanted from the 70-hole trays to 45-hole trays with pea / cocos plugs as substrate (Xcellent Plug Quick plug BV, Monster, The Netherlands).

[0207] Repotting a plant is considered a stressful period: the roots are inevitably damaged and the root support (bark) immediately after repotting often does not have optimal properties in terms of moisture retention and capillary action, which the plant experiences as drought, i.e. as a kind of abiotic stress.

[0208] After replanting, they were treated weekly with 2500-fold diluted PTSO extract (800 ml / ha). 26 weeks after replanting, 13,000 plants were found to have failed (=completely died) compared to 13,000 untreated plants. The results are shown in Table 2a below.

[0209] [Table 2a]

[0210] From these results it is clear that treatment with PTSO extract has a strong positive effect on survival rate after repotting, which can be considered as a stressful treatment.

[0211] After 26 weeks, the plants were potted into clear 12 cm pots and grown under the same cultivation regime as given in Example 2.

[0212] The hybrids Mekong, Orinoco, Ferrara and Goya were evaluated for growth as shown in Example 2, and growth enhancement was observed in all these hybrids (raw data not shown here).

[0213] After inducing flower stalks in the cold room, the number of flower stalks per plant was counted, for which 800 plants per cross were evaluated (see Table 2b below).

[0214] [Table 2b]

[0215] From Table 2b, it is clear that the number of flower stalks formed after treatment was more than that of the control group when grown under the same conditions, which makes the PTSO extract a biostimulant.

[0216] Example 3. Treatment of Chrysanthemum Cuttings

[0217] PTSO extracts were tested for their biostimulant activity as defined in the EU Regulation 2019 / 1009. The results confirmed the occurrence of the four mentioned properties on the rooting and growth of chrysanthemum cuttings. The experiments were carried out by the independent research institute Vertify (The Netherlands).

[0218] Scope: In this example, the effect of PTSO extract as a biostimulant was demonstrated. For this purpose, cuttings of Chrysanthemum variety "Chic" were selected. Chrysanthemum cuttings were planted in flat plant containers ("plots", each plot consisting of 30 plants) in standard potting soil with fertilizer.

[0219] All treatments were applied to the roots in triplicate with one week intervals as indicated below: Temperature, light control and relative humidity in the greenhouse were recorded using a climate computer (Sercom).

[0220] After treatment, the plants were evaluated at the end of the flowering induction period. The final evaluation was performed 13 days after the last treatment. The cuttings were judged for root formation, stem length, and overall condition of the plants.

[0221] Plant cell procedures

[0222] Rooted Chrysanthemum "Chic" cuttings with roots of the same length were delivered by Royal van Zanten of the Netherlands.

[0223] The trials were carried out under controlled conditions in a greenhouse facility. Wooden crates measuring 40 x 60 cm and 10 cm deep were filled with standard fertilized potting soil (organic medium) and planted with Chrysanthemum cuttings. Each plot consisted of two crates with 15 plants each (30 plants per plot).

[0224] Treatments (specific dilutions) consisted of three applications at weekly intervals, the first application was performed one week before transplanting (at the rooting stage) and the second application was performed at the time of transplanting. The last application was performed one week after transplanting. Randomization of the treatments was performed in the experimental greenhouse and the positions where the plants were placed were determined using Genstat software. Statistical analysis of the experimental results was performed using the same software. Details are described in the results section of this example.

[0225] The temperature and relative humidity in the greenhouse compartment were recorded using a climate computer and were set at 23° C. and 50%, respectively.

[0226] Application targeted to the roots was by drench or spray. The first application to the roots was done by drenching the plants in the solution for approximately 15 minutes. Untreated plots were submerged in water. The cuttings were kept in the crates with the roots drenched in water until they were transplanted into the crates containing soil. Further applications were done by spraying the solution onto the soil followed by a gentle rain to incorporate the product into the rooting zone. Application and evaluation details are summarized in Table 3 below.

[0227] [Table 3]

[0228] The treatments are summarized below in Table 4. Previcur Energy is the fungicide and Trianum-P is the biological fungicide.

[0229] [Table 4]

[0230] At each evaluation date, the plant height of 10 central plants per plot was measured. An additional evaluation of general crop vigor was made. General crop vigor was recorded on an index scale of 1 to 10 (1 = very poor crop vigor; 10 = excellent (fair or better) crop vigor) and based on the impression of general health. At the final evaluation date, the fresh weight of the above-ground plant mass was measured. The rooting density on the bottom of the crates was recorded. For this purpose, the crates were turned upside down and the percentage of roots covering the underside of the bottom was estimated.

[0231] Statistical analysis was performed using Genstat (LSD test, 95%). P in the table means probability. If P is less than or equal to 0.05, the difference between two treatments is statistically significant. The least significant difference (LSD) is the smallest difference between treatments that is significantly different at 95% (P=0.05). Values ​​indicated with the same letter are not significantly different (P=0.05).

[0232] One liter of PTSO extract was dissolved in 1,000 liters of water (dilution ratio 1:1,000) and 1,001 liters of this volume was sprayed onto a soil surface area of ​​1 ha.

[0233] result

[0234] The results after analyzing the crop height are summarized in Table 5 below.

[0235] [Table 5]

[0236] In the case of the PTSO extract with the highest concentration (1:1000), the plants were significantly lower at 0 DA-B at transplantation compared to all other treatments. At a concentration of 1:2,500, no adverse effect on plant height was observed with the drench treatment.

[0237] The differences were visible after repeated applications and over the test period. A dose-response was constructed with the PTSO extract. The tallest plants were measured at concentrations of 1:10,000 and 1:50,000.

[0238] General crop vigor was recorded on an index scale of 1 to 10 (1 = very poor crop vigor; 10 = excellent (above fair) crop vigor). Crop vigor is a subjective measurement of plant color and shape. Results are summarized in Table 6 below.

[0239] [Table 6]

[0240] For general crop vigor, only the PTSO extract at a concentration of 1:1,000 showed lower results compared to all other treatments. On the final evaluation day (19 days after transplanting), the fresh weight of the above-ground plant mass was measured. The rooting density on the bottom of the crate was recorded. The results are summarized in Table 7 below.

[0241] [Table 7]

[0242] The fresh weight of above ground plant mass of plants treated with PTSO extract at a concentration of 1:1,000 applied as a soil treatment was less than other treatments. Rooting at transplanting appeared to be affected, but final results obtained 19 days after transplanting did not show any adverse effect on rooting. In this treatment, the rooting density was even higher than other treatments. After all soil treatments with PTSO extract, the rooting density was equal (entry 7 in Table 7) or higher than both the untreated plots and the benchmarks Previcur Energy and Trianum-P (entries 4-6 and 8 in Table 7).

[0243] Few roots were observed after the first application during the rooting stage (drenching) with PTSO extract at the highest concentration (1:1,000). The plants were also shorter compared to untreated plots. Also, with PTSO extract at a concentration of 1:2,500, a smaller number of roots was observed compared to untreated and the benchmarks Previcur Energy and Trianum-P.

[0244] A dose-response relationship was observed for the treatment with PTSO extract after repeated application and during the test period after soil application. The tallest plants were measured at lower concentrations of PTSO extract, i.e., 1:10,000 and 1:50,000. At these concentrations, they were significantly taller than untreated plants until 2 weeks after planting. When exposed to the highest concentration of PTSO extract, i.e., 1:1,000 PTSO extract, the plants were smaller than all other treatments. The difference was also reflected by the fresh weight at the final evaluation day (19 days after planting).

[0245] During treatment with PTSO extract at a concentration of 1:1,000, rooting during the rooting stage (drenching) was affected, resulting in smaller plants compared to those subjected to other treatments, but no adverse effect on final rooting was observed. After treatment with PTSO extract at a concentration of 1:1,000, rooting at 19 days after transplanting was numerically better compared to treatments with other concentrations tested. Moreover, when applied as a soil treatment, PTSO extract resulted in numerically better rooting compared to untreated plots as well as to the standards Previcur Energy and Trianum-P.

[0246] Chrysanthemum is a mass produced cut flower, retailed by weight. In this example, it is clearly demonstrated that the composition provides more weight and is therefore beneficial to the horticulturist.

[0247] It is also clear from these experiments that in order to obtain a biostimulatory effect, an appropriate amount of the biostimulant must be administered.

[0248] Example 4. Treatment of lettuce seeds and seedlings

[0249] Seeds of lettuce (Lactuca sativa) variety "volare" were obtained from Enza Zaden, and Trianum P from Bayer AG, Crop Science Division.

[0250] Seeding boxes (45 pieces, 30 cm, 8 cm deep, 9 liters of soil / seeding box) were filled with standard seeding soil (BVB Substrates), consisting of a fine organic medium, in which lettuce was sown. The resulting seeding soil was fertilized by the grower and no additional fertilizer was applied during the experimental period. Each plot consisted of one seeding box with 50 seeds, planted 2 cm deep.

[0251] As shown in Table 8, seeds were proximity treated prior to sowing.

[0252] [Table 8]

[0253] A solution with PTSO extract was prepared in tap water. Application was performed by irrigating the seeds with the prepared solution for 10 minutes. After irrigation, the seeds were air-dried and sown. Untreated seeds were irrigated in water. "Vital seeds" was used as the reference drug. The boxes were covered until germination. The temperature was set at 21°C and the humidity varied from 70 to 95%. Light exposure was achieved by three 25 watt LED tubes at a distance of 30 cm from the plants (50 μmol). After germination, the lids were removed.

[0254] Randomization of treatments was performed manually. Statistical analysis of study results was performed using Genstat software. Temperature, light control and relative humidity were controlled and recorded using a climate computer (Sercom).

[0255] Study details are summarized in Table 9 below.

[0256] [Table 9]

[0257] When all seeds had fully germinated, the plants were scored as either normal, dwarf, or abnormal (plants with malformed cotyledons or leaves) and general crop vigor was recorded on an index scale of 1 to 10. On the final evaluation day (four weeks after planting the seeds), the fresh weight of the above-ground plant mass was measured.

[0258] At each evaluation date, the number of germinated plants was counted per plot. After full germination, the plants were scored into the categories of normal, small, or abnormal (plants with malformed cotyledons or leaves). Additionally, general crop vigor was recorded on an index scale of 1 to 10 (1 = very poor crop vigor; 10 = excellent (above average) crop vigor). At the final evaluation date, the fresh weight of the aboveground plant mass was measured. Statistical analysis was performed using Genstat (LSD test, 95%). In the table, P means probability. The difference between two treatments is statistically significant when P has a value of 0.05 or less. The least significant difference (lsd) is the smallest difference between treatments that is significantly different at 95% (P = 0.05). Values ​​with the same letter are not significantly different (P = 0.05). For example, at 12 DA-S, no statistically significant differences were observed between untreated lines ("a") and viable seeds ("a"), PE;1:50,000 ("a"), or PE;1:25,000 ("ab"), but statistically significant differences were observed between untreated lines ("a") and PE;1:10,000 ("bc") and PE;1:5,000 ("c"). Similarly, the PE;1:5,000 ("c") treatment was significantly different from all other treatments except PE;1:10,000 ("bc").

[0259] Results and Discussion

[0260] On each assessment date, the number of germinated seedlings was counted per plot.

[0261] When all plants had fully emerged, the plants were scored as either normal, dwarf, or abnormal (plants with malformed cotyledons or leaves).

[0262] The results are summarized in Tables 10 and 11 below.

[0263] [Table 10]

[0264] [Table 11]

[0265] It can be seen that only a slight difference was observed between the number of germinated plants and the number of normal plants, and few small or abnormal plants were observed.

[0266] During germination and at full emergence, no significant differences were found between treatments in both the number of germinated and normal plants. Germination levels were very good in all treatments. Treatments did not affect seedling emergence or quality, either positively or negatively.

[0267] At 12 days after sowing and on subsequent evaluation dates, seed shedding was observed, especially in untreated, viable seeds, and at low PE concentrations. No explanation was found. No phytopathogenic or other plaques were detected.

[0268] Significant differences were observed among treatments. A dose-response was constructed for PE. At the highest concentration (1:5,000), PE did not induce plant abscission. At the lowest concentration (1:50,000), PE was comparable to untreated plots.

[0269] At lower dilutions of PE (i.e., higher concentrations of PE), when applied as a seed drench application to seeds, seedlings were more resilient compared to seedlings from untreated seeds. Significant effects were measured at concentrations of 1:5,000 and 1:10,000. In the case of vital seeds, no effect was observed in untreated plots.

[0270] General crop vigor was recorded on an index scale of 1 to 10 (1 = very poor crop vigor; 10 = excellent (above fair) crop vigor). Crop vigor is an estimate of the plant, in this case color and shape (Table 12 below).

[0271] [Table 12]

[0272] Dilutions of 1:5,000 and 1:10,000 resulted in some more vigorous plants compared to the lower concentration of viable seeds and untreated plots.

[0273] On the final evaluation day (28 days after sowing; 24 days after germination), the fresh weight of the above-ground plant mass was measured. The results are summarized in Table 13 below.

[0274] [Table 13]

[0275] All treatments with PTSO extract dilutions resulted in an increase in fresh weight over the control. In conclusion, germination levels were very good for all treatments. Treatments did not adversely affect seedling emergence or early quality. Seedlings germinated from seeds treated with PE at 1:5,000 and 1:10,000 were significantly more resilient than seedlings germinated from untreated and viable seeds. Plants lost less leaves (±1% vs. ±10% untreated) and plant vigor was better than untreated plots. Plant weights measured 28 days after sowing were also greater for treatments with PE at 1:5,000 and 1:10,000, resulting in an 11% increase in weight over untreated and 26% over benchmark viable seeds. Plant size at that time was 4-6 grown leaves.

[0276] No effect was measured with the reference agent, Viable Seed, compared to untreated plots.

[0277] conclusion

[0278] This example demonstrated that use of PTSO extract resulted in larger lettuce heads and increased yield per nutrient input (i.e., a marker of nutrient utilization efficiency) as well as increased crop vigor, providing further evidence of the biostimulatory effects of the compounds disclosed herein.

[0279] Example 5. Treatment of tomato plants

[0280] Scope: In this example, the biostimulatory effect of PTSO extract (PE) and the method of administration were demonstrated. For this purpose, the commercial tomato cultivar "Xandor" was selected.

[0281] Plant Cultivation Procedure

[0282] Tomato cultivar "Xandor" from Axia seeds (Netherlands) was used. The tomato plants were pre-grown to a height of 60 cm.

[0283] The tomato plants of the variety "Xandor" were pre-cultured and planted in spring in rock wool mats (Grodan (ROCKWOOL BV)) in a greenhouse. The plants were fed and adjusted online according to actual evaporation and weather by a computer system provided by Priva BV. All nutrients in the plant feed were overfed and the compositions were sampled weekly and checked for nutrient limitations. EC value was maintained between 2.3 and 3.0, pH was kept at 5.4 and was continuously corrected. Table 14 below shows the growth parameters that were adjusted, monitored and controlled. Table 14 shows the settings, but the actual values ​​differ considerably due to changes in weather conditions during the day.

[0284] [Table 14]

[0285] [Table 15]

[0286] Treatments are summarized below in Table 16. Serenade is a fungicide that is often applied in tomato crops.

[0287] [Table 16]

[0288] Each treatment consisted of a group of 13 strains.

[0289] The amount of spray applied was 150ml / m 2100 ml / plant was applied on the mat. An additional assessment was made for general crop vigor. General crop vigor was recorded on an index scale of 1 to 10 (1 = very poor crop vigor; 10 = excellent (fair or better) crop vigor) and was based on an impression of general health. Crop vigor criteria included "general condition of the plant (freshness of the plant, firmness of the leaves, healthy leaves)", "tomato head quality" (head diameter, stem thickness, and rated from 1 (very poor) to 10 (excellent) and leaf color (1 (yellow) to 5 (green)).

[0290] Additionally, once a week, ripe tomatoes were harvested and weighed.

[0291] Statistical analysis was performed using Genstat (LSD test, 95%). In the table, P means probability. The difference between two treatments is statistically significant when P has a value of 0.05 or less. The least significant difference (lsd) is the smallest difference between treatments that is significantly different at 95% (P=0.05). Values ​​with the same letter are not significantly different (P=0.05).

[0292] After planting on the mats, crop development proceeded as expected for this variety. Evaluation began on day 124. The results are presented in Tables 17-20 below.

[0293] Table 17 demonstrates that the PTSO extract did not adversely affect cumulative tomato yield and slightly increased cumulative tomato yield at a dilution of 1:10,000. Many plant quality parameters were scored, such as those listed above, including tomato head quality (Table 18), plant health (Table 19) and crop color (Table 20). All dilutions of the PTSO extract showed improved quality in terms of tomato heads, plant health and plant color.

[0294] Although the fertilization conditions were the same for all plants, more plant biomass was formed in the plants treated with PTSO extract, along with improved plant characteristics, indicating that the plants treated with PTSO extract had a higher nutrient utilization efficiency. It was quite surprising that the observations made after treatment of tomato plants treated with PTSO extract during the first 4 weeks of tomato crop cultivation showed that it was sufficient to induce positive effects over a cultivation period of more than 6 months.

[0295] No abnormalities were observed in terms of wilting, flower buds and / or flower drop, shape or size of fruit, stems or leaves.

[0296] Although all treatments showed improvement in crop quality parameters, only one application method was found to be effective in increasing yield, i.e. repeated dropwise treatment with 25,000-fold diluted PTSO extract, which resulted in an 8% increase in cumulative tomato production.

[0297] In this example, the PTSO extract is clearly shown to be a powerful biostimulant, especially when applied multiple times via irrigation water.

[0298] [Table 17]

[0299] [Table 18]

[0300] [Table 19]

[0301] [Table 20]

[0302] Example 5b. Treatment of various tomato varieties

[0303] In this example, the biostimulant effect of PTSO extract (PE) was demonstrated in various varieties of tomato and with different growers.

[0304] Scope: Various varieties of tomato plants were grown by different growers and treated with PTSO extracts. The tomato varieties and growers are summarized in Table 20a below. The same tomato cultivation regime as described in Example 5 was used.

[0305] [Table 20a]

[0306] Conclusion: Treatment with PTSO extract clearly demonstrated yield increase in all tomato varieties tested and is widely applicable.

[0307] Example 6 Lettuce

[0308] Based on the results from above, an agricultural composition was prepared containing 5.2% PTSO. The composition further contains 59% emulsifiers (propylene glycol and polyethylene glycol glyceryl ricinoleate). The composition is referred to herein as "PTSO Composition 5.2".

[0309] An orchard of outdoor lettuce variety Iceberg was selected. A randomised complete block design was performed with four replicates of each treatment. Six drip applications were performed through a drip irrigation system with a pressure of 120 kPa and a water volume of 10,000 L / ha.

[0310] Treatment 1: Untreated Check + Farmer Program + MATTER ORGANIC 10L / ha Treatment 2: 0.5L / ha PTSO Composition 5.2 + Farmer Program + MATTER ORGANIC 10L / ha Treatment 3: PTSO Composition 5.2 + Farmer Program + MATTER ORGANIC 10L / ha at 1L / ha

[0311] In the examples described herein, "FARMER PROGRAM" refers to the standard growing conditions used by farmers, such as nutrients, standard crop protection, light regimes, etc.

[0312] process

[0313] The treatments were applied six times, approximately every two weeks, at intervals of 14-24 days. The final treatment is referred to as treatment "F" and the blow referred to as 10DA-F refers to the evaluation performed 10 days after the final treatment (treatment "F").

[0314] Marketable yield (number of lettuce per hectare, kilograms of lettuce per hectare (kg / ha), weight per lettuce (grams), and percentage of the harvest that was marketable) was assessed at 10DA-F. In addition, number of non-marketable lettuce per hectare and percentage of the harvest that was non-marketable were assessed at 10DA-F.

[0315] The rest of the treatments showed higher values ​​for marketable yield when treatment 1 was given a value of 100%. Treatment 3 showed the highest values ​​(102% for seeds per hectare, 121% for kilograms per hectare, 118% for average grams per seed, 102% for percentage of marketable lettuce in the harvest), followed by treatment 2 (101% for seeds per hectare, 110% for kilograms per hectare, 108% for average grams per seed, 102% for percentage of marketable lettuce in the harvest).

[0316] The rest of the treatments showed low values ​​for non-marketable yield when treatment 1 was given a value of 100%. Treatment 3 showed the lowest values ​​(61% for numbers per hectare and percentage of non-marketable lettuce in the harvest) followed by treatment 2 (69% for numbers per hectare and percentage of non-marketable lettuce in the harvest).

[0317] The laboratory trial was conducted with eight lettuce plants per plot at similar developmental stages, and one evaluation was performed at 10 DA-F. The aboveground fresh weight (g), root weight (g), firmness (kg / cm 2 ) and actual diameter (cm) were assessed.

[0318] Regarding top fresh weight and root fresh weight, the rest of the treatments showed higher values ​​when treatment 1 was given a value of 100%. The highest top and root fresh weights were obtained in treatment 3 (117% and 123%, respectively), followed by treatment 2 (107% and 101%, respectively).

[0319] For lettuce diameter, when treatment 1 was given a value of 100%, treatment 3 had a value of 107% and treatment 2 had a value of 104%.

[0320] With regard to lettuce firmness, when treatment 1 was given a value of 100%, treatment 3 had a value of 133% and treatment 2 had a value of 125%.

[0321] Finally, five lettuce heads per plot at similar developmental stages from each harvest were removed, weighed, placed in a refrigerated room at 6°C, and the weight loss per head was assessed over the next 13 days; 92 DAP (Days After Plant), 93 DAP, 94 DAP, 95 DAP, 96 DAP, 97 DAP, 98 DAP, 99 DAP, 100 DAP, 101 DAP, 102 DAP, 103 DAP, and 104 DAP (the percentage weight loss per head over the entire storage period was calculated). The same lettuce were then left at room temperature (14°C) and the weight loss per lettuce re-evaluated over the next 7 days; 105 DAP, 106 DAP, 107 DAP, 108 DAP, 109 DAP, 110 DAP and 111 DAP (the final percentage of weight lost per lettuce over the entire storage period was calculated).

[0322] Example 7 Cucumber

[0323] Cucumber variety Katrina orchards were selected. The basic plot is 18 m 2 With a water volume of 10,000 L / ha and a pressure of 120 kPa, 14 drip applications (applications ABCDEFGHIJKLMN) were administered for PTSO composition 5.2 and 13 plants per plot. 商標 Seven drip applications (ACEGIKM) were made using a drip irrigation system. The applications were made at 7-day intervals for PTSO Composition 5.2 and Isabion 商標 were performed at 14-day intervals.

[0324] Process 1: Unprocessed check Process 2: Farmer check Treatment 3: 0.4 L / ha PTSO composition 5.2 + 4 L / ha Isabion 商標 +41g free amino acids, 40g nitrogen, 118g organic carbon Treatment 4: 0.4 L / ha of PTSO composition 5.2

[0325] Marketable yield (kilograms of fruit per hectare (kg / ha), number of fruit per hectare, and fruit weight (g)) was evaluated for 3DA-B (i.e., 3 days after treatment B), 6DA-B, 3DA-C, 6DA-C, 3DA-D, 6DA-D, 3DA-E, 7DA-E, 4DA-F, 1DA-G, 5DA-G, 3DA-H, 6DA-H, 3DA-I, 7DA-I, 5DA-J, 3DA-K, 7DA-K, 4DA-L, 1DA-M, 5DA-M, and 3DA-N by calculating the percentage weight loss per lettuce over the entire storage period, and finally the total yield and the average value were calculated. Fruit diameter and length (mm) were evaluated for 3DA-E and 7DA-K.

[0326] In terms of marketable yield (kg of fruit per hectare and number of fruit per hectare), when treatment 1 was given a value of 100%, treatment 3 obtained the highest yield (116% in kg / ha and 113% in number of fruit), followed by treatment 4 (112% in kg / ha and 110% in number of fruit), and then treatment 2 (104% in kg / ha and 104% in number of fruit).

[0327] Regarding fruit diameter, when treatment 1 was given a value of 100%, treatment 3 gave results of 103-104%, treatment 4 gave results of 102-104%, and treatment 1 gave results of 102-103%.

[0328] Regarding fruit length, when treatment 1 was given a value of 100%, treatment 3 gave results of 106–107%, treatment 4 gave results of 106%, and treatment 1 gave results of 103–105%.

[0329] Example 8 Treatment of cloned Cymbidium strains

[0330] Growers of Cymbidiums that produce cut flowers usually have to grow their own plants before the flowers can begin to bloom. To obtain flowers that are as uniform as possible, they must pre-grow their plants from clones (meristems). It may take five years or more before flower production reaches an economically desirable level.

[0331] Plant growth and development involves three stages: (1) vegetative growth, where pseudobulbs are formed at the end of shoot formation, (2) flower induction, and (3) flower stalk development. It is economically interesting to keep the growth time as short as possible to allow plants to grow to flowering size. In this experimental design, PTSO extract (at least 56% emulsified PTSO (PE)) is used to demonstrate the biostimulatory effect of PTSO on young plants of Cymbidium.

[0332] For this purpose, a young dividing plant of the orchid family Cymbidium "49er" was selected. The plant had not yet developed the first rhizoids after being transplanted from the flask. After the treatments described below, the plant was evaluated after 206 days as shown below.

[0333] Experimental design

[0334] Plant Cultivation Procedure

[0335] A plant of Cymbidium "49er" was created from the meristem. Upon delivery to the Cymbidium grower, the plant was fertilized with Osmocote granules (1.25 kg / m2). 3 The plants were potted in 9 cm black pots in medium sized coco peat with added fertilizer (EC 0.4) twice a day at 40 ml via an automatic watering system via an injection system. After adaptation to greenhouse conditions, treatment with PTSO extracts was initiated approximately 4 months after repotting.

[0336] In this example, 25 plants were used per treatment. The spacing between plants was about 15 cm. The plants were monitored for leaf growth by measuring the length and maximum width of the largest leaf, rhizoid formation, final size and appearance, and number of shoots. The number of roots and green tips of the roots were visually monitored. Additionally, an overall judgment was made on the plants regarding appearance and health. 8 to 24 plants were judged per treatment.

[0337] The dilution, frequency and intervention time of repeated treatment of PTSO extract are shown in Table 21 below. Plants were manually fed with 40ml of PTSO extract of the indicated dilution. Controls were fed with 40ml of standard fertilized plant feeding as described above. After treatment, the treated plants were subjected to the same cultivation procedures as the control plants, i.e., untreated plants.

[0338] [Table 21]

[0339] The lines were scored for the indicated variables at t=0, t=112 days, and t=206 days.

[0340] The height and thickness of the rhizoids; length; and width were measured by using a tape-line. The amount of root and the fresh white growing root tip were estimated on a scale of 1 to 10.

[0341] The amount of roots was determined after comparing the roots of the plant with those of a plant with average root formation of the control group. For this purpose, the plant was removed from the pot and compared with a standard plant, which was judged as the "average" of the control treatment.

[0342] The number of fresh root tips was scored as follows: 1: 0-20% of roots showed white grown root tips; 2: 20-40% of roots showed white grown root tips; 3: 40-60% of roots showed fresh grown root tips; 4: 60-80% of roots showed grown white root tips; 5: 80-100% of roots showed grown white root tips. "Shoot emergence" was calculated as the number of shoots divided by the total number of scored plants in the treatment.

[0343] The "Robustness" of a strain is an arbitrary judgment given after the strain is compared to the average strain in a control group and ranges from 1 to 10, with 1 being the lowest score and 10 being the highest score.

[0344] result

[0345] In Tables 22-24 the average results of the treatments are presented.

[0346] [Table 22]

[0347] [Table 23]

[0348] [Table 24]

[0349] During the experiment, all plants developed well, and no adverse effect of PTSO extract on the strain was detected. The strains were ranked at t=0 (Table 2), but strictly speaking, the difference between the strains was very small. This is normal for plants that grow from meristems and are cultivated under the same conditions for 4 months before the start of the experiment. Hereinafter, the strains were treated according to the schedule presented in Table 21.

[0350] After 112 days, plants from treatment 11 were significantly larger and more robust (Table 23). High scores indicate well-developed and robust plants with relatively large rhizoids (for reserve carbohydrate storage), large leaf areas (for photosynthesis), large fast-growing shoots, and well-developed root systems for water and mineral uptake. Repeated application of PTSO extract resulted in more positive effects (see, e.g., treatments 7-11).

[0351] Example 9. Effects of organosulfur compounds on lettuce and sweet peppers

[0352] Experimental design: The biostimulatory effect of organosulfur compounds was tested on lettuce and sweet pepper (Capsicum annuum "Ritmico"). The compounds tested included di-n-propyl disulfide (CAS number 629-19-6), di-n-propyl thiosulfonate (i.e., PTSO, CAS number 1113-13-9), dimethyl thiosulfonate (CAS number 2949-92-0), and diphenyl thiosulfonate (Cas number 1212-08-4). Dimethyl thiosulfonate and diphenyl thiosulfonate were obtained from Adrich-Sigma. PTSO was purified as described in Example 1. To obtain a stable and homogeneous emulsion, the compounds were dissolved in an emulsifier solution (Table 25) and the crop was fed with the defined solution. In this way, the occurrence of precipitation or immiscibility of the organosulfur compounds was prevented. As examples herein, "PTSO Composition 5.2" refers to an agricultural composition that contains 5.2% PTSO and further contains 59% emulsifiers (propylene glycol and polyethylene glycol glyceryl ricinoleate). "PTSO Composition 5.2 Yuka" in the agricultural composition contains 5.2% PTSO and 90.2% Yuka extract. "PTSO Composition 5.2 Tween" in the agricultural composition contains 5.2% PTSO, 30.1% Tween, 30.1% DMSO and 30% water. Controls and treatments are shown in Table 27 below.

[0353] The amount of test compound given was standardized based on molar amount, and the same molar amount was given as PTSO. Emulsions were prepared by adding the required amount of compound as shown in Table 25 to 100 ml of 50% Tween 80 and 50 ml of DMSO. They were then made up to a final volume of 250 ml with the same mixture and incubated on an orbital shaker at 37°C (30 min, 250 rpm).

[0354] [Table 25]

[0355] Before sowing, lettuce seeds were immersed in 1000-fold diluted emulsions as shown in Table 25 and incubated for 10 minutes. Each treatment was performed in quadruplicate. In Table 26, the growth settings for lettuce and sweet peppers are summarized, and in Table 27, an overview of the achieved growth conditions and harvest for lettuce is presented.

[0356] As shown in Table 27, all lettuce treatments showed an increase in relative fresh weight / plant compared to the control, and an increase in harvest weight per plant compared to the control. At the time sweet pepper growth was measured, no fruits were present. However, preliminary results showed that all treatments resulted in an increase in crop vigor as measured by relative size compared to the control.

[0357] [Table 26]

[0358] [Table 27]

[0359] Example 10. Analysis of PTSO in onions and garlic oil

[0360] Scope: Onion and garlic oils and extracts of onion and garlic were obtained as described in Hemat S.Abd El-Salam et al.(2014) I Enhancement of Cumin (Cuminum cyminum L.) Productivity Using Some Natural Plant Extracts.Egypt.J.Hort.Vol.41,No.2,p 209-219. Samples were analyzed for the presence of PTSO as follows:

[0361] Experimental design: The presence of PTSO (di-n-propylthiosulfonate) was detected by LC-MS / MS. A standard substance of PTSO (CAS number 1113-13-9) was obtained by organic synthesis from Symeres (Mercachem Holding BV). The identity of the compound was confirmed by 1 It was confirmed by 1 H-NMR and LC-MS, and the purity was determined by LC-UV and was determined to be greater than 99.1%.

[0362] This sample was used to generate the equilibration graph. Samples containing 2-200 μM pure PTSO in HPLC grade methanol were prepared for spiking and equilibration.

[0363] The LC-MS / MS (API4000, Sciex) had the following configuration: synchronous mode setting LC-Sync, a Vanguard BEH C18 2.1 x 5 mm precolumn, and an Acquity BEH C18 2.1 x 50 mm 1.7 μm HPLC column were used. The injector was supplied by Nexer.

[0364] The oven was set to 40°C. The pump mode was ternary flow. The pump settings were as follows: total flow rate was 0.4 ml / min; concentration (%) for pump B was 0.0 and concentration (%) for pump C was 50.0. Pump B and pump C had zero curves and minimum and maximum pressure limits of 0 and 16,000 psi, respectively. Pump A was used with solution LC-A (0.05% FA in 5% methanol in ultrapure water). Pump C was used to deliver methanol.

[0365] MS settings:

[0366] [Table 28]

[0367] [Table 29]

[0368] Methods: Calibration standards in the range of 2.00-200 μM (PTSO) were freshly prepared in MeOH.

[0369] Isolation of PTSO from samples was performed by extraction into organic solvents (methanol (MeOH), ethyl acetate (EtOAc), n-butanol and dichloromethane (DCM)). The sample to extract ratio was up to 1. The results of the preparative study showed that the extraction efficiency of the samples was 30-40%, therefore, a reference sample needs to be included in the procedure.

[0370] A sample volume of 20 μl was injected by the autosampler.

[0371] The extracts were analyzed using an API 4000 LC-MS / MS system (PTSO).

[0372] Data acquisition was performed using Analyst software (version 1.6.3) from AB Sciex. After integration of peak areas, regression was also performed using Analyst. Concentrations were calculated using weighted linear regression according to the following formula: y=a+bx (weighting factor=1 / x2), where x=PTSO concentration (μM), y=peak area ratio, a=intercept, and b=slope.

[0373] The calibration curve range for PTSO is 2.0-200 μM, the quantitative analysis range is greater than 2.0 μM, and the qualitative analysis range is greater than 1.0 μM, with a sample volume of 20 μl in each case.

[0374] result:

[0375] [Table 30]

[0376] These results demonstrate that no PTSO was detected in onion and garlic from the individual oils and extracts.

[0377] Example 11. Effects on other crops

[0378] Scope: The effect of the PTSO extract has been tested on a variety of other crops. Table 31 below lists the results from crops where a visible effect in at least one measured variable was observed.

[0379] Study Design: Table 32 below summarizes the crops tested and their corresponding growing conditions and treatments. The crops were pre-grown in plant growers and then planted in greenhouses with constant aeration (day and night) or in open fields. Plants were given irrigation water or via plugs. All nutrients in the plant feed were provided in excess. Depending on the crop, fruits were harvested at regular intervals and the cumulative weight of the fruits was determined. PTSO extract was provided via the roots via plant feed. In a limited number of crops, root formation and chlorophyll content were also measured.

[0380] In table 32 below, the monitored growth variables and yields are presented. There is a high variability in the growth parameters temperature, relative humidity and light supply, because these parameters are not controlled and depend on the weather, which changes during the day. Weather conditions change with the seasons, and therefore the month in which the experiment started is indicated in the table. June has the highest light intensity and the highest variation in day and night temperature, corresponding relative humidity and evaporation. These (extreme) variations result in high abiotic stress. Since PTSO extract is thought to reduce the sensitivity of plants to abiotic stress, it was expected that the greater the variation in the abiotic stress factors, the more pronounced the positive effect of the treatment on the crop.

[0381] Table 33 below demonstrates that the PTSO extract did not have any negative effect on the crop characteristics, such as damaged leaves, crop color, deformed leaves or size, stems, fruit shape and flowers, and also did not have any negative effect on root formation. The crop condition remained unchanged or improved.

[0382] The growth conditions were kept the same for the plants treated with PTSO extract and the untreated plants (control), which means that the nutrient utilization efficiency for the plants treated with PTSO extract was higher, thus proving that PTSO is a biostimulant.

[0383] Since the ventilation of the greenhouse is constant during the day and is not controlled as a function of temperature, large deviations in temperature and humidity are expected depending on the wind and sun. The relative air humidity varies inversely with the temperature, therefore large deviations are also expected. The supply of irrigation water may also be limited during hot periods, resulting in too little water uptake, which relaxes the turgor pressure of the cells, causes the leaves to droop and the stomata to close. At that moment, photosynthesis stops. These uncontrolled environmental factors result in many abiotic stresses and, without wishing to be bound by theory, it has been suggested that the PTSO extract reduces the impact of biotic stresses on the plants, resulting in higher levels of photosynthesis-related metabolites, healthier crops and higher crop yields. Depending on the crop, further positive properties were observed, such as improved root systems and improved chlorophyll levels (Table 33). Other crops experienced significantly faster growth and improved crop characteristics (roses: thicker and / or longer stems, and shinier leaves; melons: greener and larger leaves; string beans: significantly larger crop; tomatoes: larger heads and more chlorophyll).

[0384] Table 31 provides a summary of the improved crop characteristics after treatment with PTSO extract. Not all possible crop characteristics were monitored, therefore other characteristics may have been improved but were not measured in this experiment.

[0385] [Table 31]

[0386] [Table 32] JPEG2024540856000046.jpg255140JPEG2024540856000047.jpg25566

[0387] [Table 33] JPEG2024540856000049.jpg255110

Claims

1. A biostimulant for plants comprising a compound selected from the group consisting of di-n-propylthiosulfonate (PTSO), diphenylthiosulfonate, and di-n-propylthiosulfinate (PTS).

2. The biostimulant, a) nutrient use efficiency, b) tolerance to abiotic stress, and / or c) quality attributes The biostimulant of claim 1, which is used to improve one or more of the following:

3. A method comprising providing a plant with a compound selected from the group consisting of di-n-propylthiosulfonate (PTSO), diphenylthiosulfonate, and di-n-propylthiosulfinate (PTS), or a composition containing a compound selected from the group consisting of PTSO, diphenylthiosulfonate, and PTS.

4. 3. The biostimulant according to claim 1 or 2, wherein the biostimulant is provided to the plant at least 6 times during the crop cycle of the plant, and / or the biostimulant is provided to the plant every 4 to 27 days during the crop cycle of the plant.

5. A biostimulant described in any one of claims 1 to 2, wherein the compound is provided in an amount of 0.01 kg / ha to 100 kg / ha.

6. A biostimulant described in any one of claims 1 to 2, wherein the biostimulant is provided to the plant as a solution containing the compound in an amount of 0.5 mg / L to 150 mg / L.

7. The biostimulant according to any one of claims 1 to 2, wherein the plant belongs to the Embryophyta clade or the Angiospermae clade, preferably wherein the plant is selected from the group consisting of Phalaenopsis, Cymbidium, Chrysanthemum, Rosa, Fabaceae, Brassica, Cucurbita, Solanaceae, Pisum, Vitis, Vaccinia, and Lactuca.

8. The biostimulant according to any one of claims 1 to 2, wherein the biostimulant is applied directly to the plant, to the seeds of the plant, or to the soil of the plant or the seeds.

9. 9. The biostimulant of claim 8, wherein the biostimulant is applied via drip irrigation.

10. The biostimulant according to any one of claims 1 to 2, wherein the biostimulant is used in combination with an amino acid-based biostimulant.

11. An agricultural composition comprising a compound selected from the group consisting of di-n-propylthiosulfonate (PTSO), diphenylthiosulfonate, and di-n-propylthiosulfinate (PTS), and 40 to 70% of an emulsifier.

12. The agricultural composition of claim 11, wherein the emulsifier is selected from one or more of propylene glycol, glyceryl polyethylene glycol ricinoleate, yuca extract, and Tween.

13. A biostimulant described in any one of claims 1 to 2, wherein the compound is di-n-propylthiosulfonate (PTSO).

14. The biostimulant according to any one of claims 1 to 2, further comprising a fertilizer, an insecticide, a wetting agent, an antimicrobial compound, a fungicide, a chelating compound, an aromatic compound, and / or an additional biostimulant.

15. The biostimulant according to any one of claims 1 to 2, wherein the biostimulant comprises an emulsifier.

16. The biostimulant of any one of claims 1 to 2, wherein the biostimulant comprises an emulsifier selected from one or more of propylene glycol, glyceryl polyethylene glycol ricinoleate, yuca extract, and Tween.

17. The biostimulant according to any one of claims 1 to 2, wherein the biostimulant comprises an amino acid-based biostimulant and / or free amino acids and peptides.

18. The method of claim 3, wherein the method is for increasing the growth rate, development, yield, harvest, and / or vigor of a plant, and / or the method improves a) nutrient utilization efficiency, b) resistance to abiotic stress, and / or c) quality characteristics.

19. The method described in claim 3 or 18, wherein the compound or composition is provided to the plant at least 6 times during the plant's crop cycle, and / or the compound or composition is provided to the plant every 4 to 27 days during the plant's crop cycle.

20. The method of claim 3 or 18, wherein the compound is provided in an amount of 0.01 kg / ha to 100 kg / ha.

21. The method of claim 3 or 18, wherein the compound is provided to the plant as a solution containing the compound in an amount of 0.5 mg / L to 150 mg / L.

22. The method of claim 3 or 18, wherein the compound or composition is applied directly to the plant, to the seeds of the plant, or to the soil of the plant or the seeds.

23. The method of claim 22, wherein the compound or composition is administered via drip irrigation.

24. The method of claim 3 or 18, wherein the compound is di-n-propylthiosulfonate (PTSO).

25. The method of claim 3 or 18, wherein the composition contains an emulsifier.

26. The method of claim 25, wherein the emulsifier is selected from one or more of propylene glycol, glyceryl polyethylene glycol ricinoleate, yuca extract, and Tween.

27. ​​An agricultural composition as described in claim 11 or 12, wherein the compound is di-n-propylthiosulfonate (PTSO).

28. The agricultural composition of claim 11 or 12, wherein the composition further comprises a fertilizer, an insecticide, a wetting agent, an antimicrobial compound, a fungicide, a chelating compound, an aromatic compound, and / or an additional biostimulant.

Images