A METHOD FOR IMPROVING THE HEALTH OF A PLANT USING AT LEAST ONE (THIO)PHOSPHORIC ACID TRIAMIDE SUCH AS N-(N-BUTYL)THIOPHOSPHORIC ACID TRIAMIDE (NBPT) AND/OR N-(N-PROPYL)THIOPHOSPHORIC ACID TRIAMIDE (NPPT) ESSENTIALLY IN ABSENCE OF A UREA-CONTAINING FERTILIZER

Abstract

A method for improving the health of a plant, comprising treating a plant growing on soil or soil substituents and/or treating the locus where the plant is growing or is intended to grow with at least one (thio)phosphoric acid triamide according to the general formula (I) wherein X is oxygen or sulfur, R1 and R2 are—independent from each other—H, substituted or unsubstituted 2-nitrophenyl, C1 to C20 alkyl, C3 to C20 cycloalkyl, C6 to C20 heterocycloaryl, C6 to C20 aryl, or dialkylaminocarbonyl group wherein R1 and R2 together with the nitrogen atom linking them define a 5- or 6-membered saturated or unsaturated heterocyclic radical, which optionally comprises 1 or 2 further heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, wherein improved plant health is determined by —increased crop biomass or crop yield, —improved plant vigor, —improved plant quality, and/or —improved tolerance or resistance of the plant to abiotic stress factors.

Claims

1. A method for improving the health of a plant, comprising treating a plant growing on soil or soil substituents and/or treating the locus where the plant is growing or is intended to grow with at least one (thio)phosphoric acid triamide, wherein the at least one (thio)phosphoric acid triamide is a mixture comprising N-(n-butyl) thiophosphoric acid triamide (NBPT) and N-(n-propyl) thiophosphoric acid triamide (NPPT) wherein improved plant health is determined by increased crop biomass or crop yield, improved plant vigor, improved plant quality, and/or improved tolerance or resistance of the plant to abiotic stress factors; and wherein no urea-containing fertilizer is used.

2. The method according to claim 1, wherein the health of a plant is improved by treating a plant growing on soil or soil substituents and/or treating the locus where the plant is growing or is intended to grow with at least one (thio)phosphoric acid triamide, wherein the at least one (thio)phosphoric acid triamide is a mixture comprising N-(n-butyl) thiophosphoric acid triamide (NBPT) and N-(n-propyl) thiophosphoric acid triamide (NPPT) wherein improved plant health is determined by increased crop biomass or crop yield, improved plant vigor, improved plant quality, and/or improved tolerance or resistance of the plant to abiotic stress factors; and wherein no urea-containing fertilizer is used.

3. The method according to claim 1, wherein the plant, or the plant growing on soil or soil substituents and/or the locus where the plant is growing or is intended to grow is treated with the at least one (thio)phosphoric acid triamide in an amount of from 0.03 kg/hectare to 0.5 kg/hectare.

4. The method according to claim 1, wherein the crop biomass or crop yield is increased by at least 1%, compared to the biomass or yield of the same crop produced under the same conditions, but without the treatment of the invention.

5. The method according to claim 1, wherein improved plant vigor is determined by at least one of the features or parameters selected from the group consisting of: improved vitality of the plant, improved plant growth, improved plant development, improved visual appearance, improved plant stand, less plant verse/lodging, improved emergence, enhanced root growth and/or more developed root system, enhanced nodulation, bigger leaf blade, bigger size, increased plant weight, increased plant height, increased tiller number, increased number of side shoots, increased number of flowers per plant, increased shoot growth, increased root growth, enhanced photosynthetic activity, increased stomatal conductance, increased CO2 assimilation rate, enhanced pigment content, flowering, earlier fruiting, earlier and improved germination, earlier grain maturity, improved self-defense response, less non-productive tillers, less dead basal leaves, less input fertilizer or water needed, greener leaves, complete maturation under shortened vegetation periods, less fertilizers needed, less seeds needed, easier harvesting, ripening, longer shelf-life, longer panicles, delay of senescence, stronger and/or more productive tillers, better extractability of ingredients, improved quality of seeds for being seeded in the following seasons for seed production, altered or reduced production of ethylene and/or the inhibition of its reception by the plant, and growth repression.

6. The method according to claim 1, wherein improved plant quality is determined by at least one of the features or parameters selected from the group consisting of: increased nutrient content, increased protein content, increased content of fatty acids, increased metabolite content, increased carotenoid content, increased sugar content, increased content of amino acids, including essential amino acids, improved nutrient composition, improved protein composition, improved composition of fatty acids, improved metabolite composition, improved carotenoid composition, improved sugar composition, improved amino acids composition, im-proved or optimal fruit color, improved leaf color, higher storage capacity, and higher processability of the harvested products.

7. The method according to claim 1, wherein improved tolerance or resistance of the plant to abiotic stress factors is determined by the improved tolerance and/or resistance to at least one of the stress factors selected from the group consisting of: heat stress, temperature conditions causing heat damage to a plant, cold stress, periods of thawing and freezing, frost, variations in temperature, temperature unusual for the season, drought stress, exposure to cold water, flood, water-logging, wind, sun light, acid or alkaline pH conditions in the soil with pH values lower than pH 5 and/or pH values higher than 9, salt stress, inorganic pollution, soil contamination or soil pollution with chemicals, acid rain, air pollution, radiation, oxidative stress, organic pollution, oil and/or fuel dumping or spilling, nuclear radiation, contact with sewage, over-fertilization, nutrient deficiencies, herbicide injuries, plant wounding, compaction, natural disasters, and combinations thereof.

8. The method according to claim 5, wherein at least one of the features or parameters is improved or increased by at least 1%, compared to the plant produced under the same conditions, but without the treatment of the invention.

9. The method according to claim 7, wherein the tolerance or resistance of the plant to at least one of the stress factors is improved or increased by at least 1%, compared to the plant produced under the same conditions, but without the treatment of the invention.

10. (canceled)

Description

EXPERIMENTAL PART

[0097] Test of formulations comprising at least one (thio)phosphoric acid triamide according to the present invention on corn plant development. Preferably, the at least one (thio)phosphoric acid triamide is NBPT and/or NPPT.

[0098] More preferably, Limus® formulations on corn plant development are tested.

Side Area

[0099] Projected area (in mm2) of plant biomass (side RGB image segment above pot upper border, different from background or other selected colour). Average of 6 side images from different angles.

Side Gravity Y

[0100] Side gravity Y is a robust indication of the height of the plant. It is the distance (in mm) along y-axis from bottom of image (pot upper border) to centre of gravity of plant biomass (side RGB image segment above pot upper border, different from background or other selected colour). Average of 6 side images from different angles.

Side Greenness

[0101] The index used the average greenness of all plant biomass pixels (identified as plants). The index is based on the RGB color model (value between 0 and 255 for each color) and is calculated as the ratio of Green value over Red value. Average of 6 side images from different angles.

Side Stem Width

[0102] Average width (in mm) across the length of the plant stem. The plant stem is segmented from the side RGB image as the 2 most parallel edges in the lower plant part (limited by the last fully expanded leaf, where parallel edge starts to divert). Average of 6 side images from different angles.

Top Area

[0103] Projected area (in mm2) of plant biomass (top RGB image segment different from background or other selected colour). Value of one top RGB image.

Root Area

[0104] Projected area (in mm2) of root biomass (bottom RGB image of transparent pot, segment different from substrate background). Value of one bottom RGB image.

Corn Hybrids Used in the Experiments:

[0105] Kalimnos (KWS) [0106] Torres (KWS)

Substrates Used in the Experiments:

[0107] Organic: a mixture of organic matter (81%) and clay [0108] Mineral: 66.7% Vermiculite supplemented with sand, clay and organic matter

Camera Technology Used for Phenotyping

[0109] Root area: Bottom view RGB camera [0110] Side Greenness, Side stem width, Side Gravity Y: Side view RGB camera [0111] Top area: Top view Multispectral camera

[0112] The following table shows the air humidity and the temperature change during the 24 hours of the day at the various time points:

TABLE-US-00001 TABLE 1 Overview on the formulations: Hour (time point) air humidity in % Temperature ° C. 1 85 18 2 85 18 3 85 18 4 85 18 5 85 18 6 80 18 7 80 19 8 80 20 9 70 21 10 70 22 11 70 23 12 70 24 13 70 25 14 60 26 15 60 27 16 60 28 17 60 28 18 60 28 19 60 27 20 70 25 21 70 23 22 80 21 23 80 19 24 85 18

[0113] All percentages are weight percentages.

TABLE-US-00002 TABLE 1 part 1 D/P/PG + D/P/PG + D/P/PG + Name D/P/PG NBPT + NPPT NBPT NPPT NBPT 0.00% 18.9% 20.2% 0.00% NPPT 0.00% 6.3% 0.00% 7.8% DMSO 27.0% 20.2% 21.6% 24.9% Polyethyleneimine 8.5% 6.3% 6.7% 7.8% Propylene glycol 64.5% 48.2% 51.5% 59.5% Density 1.1 g/ccm 1.1 g/ccm 1.1 g/ccm 1.1 g/ccm

TABLE-US-00003 TABLE 1 part 2 DMSO + DMSO + DMSO + Name DMSO NBPT + NPPT NBPT NPPT NBPT 0.00% 18.9% 20.2% 0.00% NPPT 0.00% 6.3% 0.00% 7.8% DMSO 100.00% 74.8% 79.8% 92.2% Density 1.1 g/ccm 1.1 g/ccm 1.1 g/ccm 1.1 g/ccm

[0114] (“D/P/PG” stands for DMSO+polyethyleneimine+propylene glycol)

[0115] The polyethyleneimine used (see Table 1) was polyethyleneimine with a weight average molecular weight of 800 g/mol as measured by GPC (dry substance, at pH 4.5).

[0116] 2 screens: well-watered & mild nutrients (2.5 g osmocote/L; Osmocote® Exact Standard 3-4M)

[0117] Application—Drench: 6 mL at DAS00 (days after seeding)

[0118] 40 reps per treatment; randomized block design

[0119] Parameters: Root area, Side area, Side gravity Y, Side greenness, Side stem width, Top area (determined as described above)

TABLE-US-00004 TABLE 2 overview on the treatments: # Treatment 1 Treatment 2 Focus 3 D/P/PG + NBPT + NPPT D/P/PG a.i. effect 4 DMSO + NBPT + NPPT DMSO a.i. effect 8 D/P/PG + NBPT D/P/PG a.i. effect 9 DMSO + NBPT DMSO a.i. effect 10 D/P/PG + NPPT D/P/PG a.i. effect 11 DMSO + NPPT DMSO a.i. effect

TABLE-US-00005 TABLE 3 a.i. effect 3 + 4 Time better Parameter Substrate Screen Variety Point Treatment than Significance Percent Root mineral Mild Torres 7 DMSO + DMSO true 5 area Nitrogen NBPT + NPPT Deficiency Root organic Mild Kalimnos 5 DMSO + DMSO true 18 area Nitrogen NBPT + NPPT Deficiency Root organic Mild Kalimnos 5 D/P/PG + D/P/PG true 17 area Nitrogen NBPT + NPPT Deficiency Root organic Mild Kalimnos 7 D/P/PG + D/P/PG true 16 area Nitrogen NBPT + NPPT Deficiency Root organic Mild Kalimnos 8 DMSO + DMSO true 10 area Nitrogen NBPT + NPPT Deficiency Root organic Mild Kalimnos 8 D/P/PG + D/P/PG true 16 area Nitrogen NBPT + NPPT Deficiency Side mineral Mild Torres 7 DMSO + DMSO true 3 gravity Nitrogen NBPT + NPPT Y Deficiency Side mineral Mild Torres 8 DMSO + DMSO true 4 gravity Nitrogen NBPT + NPPT Y Deficiency

TABLE-US-00006 TABLE 4 a.i. effect 8 + 9 Time better Parameter Substrate Screen Variety Point Treatment than Significance Percent Side Mineral Mild Kalimnos 8 D/P/PG + NBPT D/P/PG True 3 gravity Nitrogen Y Deficiency Side Mineral Mild Torres 8 DMSO + NBPT DMSO True 4 gravity Nitrogen Y Deficiency Side mineral Mild Torres 7 DMSO + NBPT DMSO True 4 gravity Nitrogen Y Deficiency Side mineral Mild Torres 8 DMSO + NBPT DMSO true 5 area Nitrogen Deficiency Side mineral Mild Torres 7 DMSO + NBPT DMSO true 6 area Nitrogen Deficiency Root organic Mild Kalimnos 8 DMSO + NBPT DMSO true 12 area Nitrogen Deficiency Root organic Mild Kalimnos 7 DMSO + NBPT DMSO true 15 area Nitrogen Deficiency Root organic Mild Kalimnos 5 DMSO + NBPT DMSO true 21 area Nitrogen Deficiency Side mineral Mild Kalimnos 8 DMSO + NBPT DMSO true 4 stem Nitrogen Width Deficiency

TABLE-US-00007 TABLE 5 a.i. effect 10 + 11 Time better Parameter Substrate Screen Variety Point Treatment than Significance Percent Side organic Mild Kalimnos 8 D/P/PG + NPPT D/P/PG True 5 gravity Nitrogen Y Deficiency Side organic Mild Kalimnos 7 D/P/PG + NPPT D/P/PG True 5 gravity Nitrogen Y Deficiency Side organic Mild Torres 7 DMSO + NPPT DMSO True 3 gravity Nitrogen Y Deficiency Side organic Mild Torres 8 DMSO + NPPT DMSO True 4 gravity Nitrogen Y Deficiency Root organic Mild Kalimnos 8 D/P/PG + NPPT D/P/PG true 13 area Nitrogen Deficiency Root organic Mild Kalimnos 7 D/P/PG + NPPT D/P/PG true 13 area Nitrogen Deficiency Root organic Mild Kalimnos 8 DMSO + NPPT DMSO true 13 area Nitrogen Deficiency Root organic Mild Kalimnos 7 DMSO + NPPT DMSO true 15 area Nitrogen Deficiency Root organic Mild Kalimnos 5 DMSO + NPPT DMSO true 24 area Nitrogen Deficiency

[0120] The experimental results show that the health of a plant could be improved via the method of the invention. The percentage values in Table 3, Table 4, and Table 5, last column, show how much the plant-health-related parameters have been improved.

Statistics:

[0121] Traits were modelled individually using a Linear Mixed Effect Model (LMM).

[0122] Treatment, Time and their interaction where used as fixed categorical explanatory variables.

[0123] The distribution of the trait was assumed to be normal and we fitted varying variances for each timepoint.

[0124] Block and Plant within Block where used as random intercept, to capture the grouping structures in the experimental design.

[0125] Post-hoc pairwise & additional custom contrasts for treatments were computed for each timepoint separately.

[0126] In a preferred embodiment, the present invention refers to the following further items. [0127] 1. A method for improving the health of a plant, comprising treating a plant growing on soil or soil substituents and/or treating the locus where the plant is growing or is intended to grow with at least one (thio)phosphoric acid triamide according to the general formula (I)

R.sup.1R.sup.2N—P(X)(NH.sub.2).sub.2  (I) [0128] wherein [0129] X is oxygen or sulfur; [0130] R.sup.1 and R.sup.2 are—independent from each other—H, substituted or unsubstituted 2-nitrophenyl, C.sub.1 to C.sub.20 alkyl, C.sub.3 to C.sub.20 cycloalkyl, C.sub.6 to C.sub.20 heterocycloaryl, C.sub.6 to C.sub.20 aryl, or dialkylaminocarbonyl group, wherein R.sup.1 and R.sup.2 together with the nitrogen atom linking them define a 5- or 6-membered saturated or unsaturated heterocyclic radical, which optionally comprises 1 or 2 further heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, [0131] wherein improved plant health is determined by [0132] increased crop biomass or crop yield, [0133] improved plant vigor, [0134] improved plant quality, and/or [0135] improved tolerance or resistance of the plant to abiotic stress factors. [0136] 2. The method according to item 1, wherein no urea-containing fertilizer is used or wherein less than 0.03 kg/hectare of urea-containing fertilizer is used. [0137] 3. The method according to item 1 or 2, wherein the at least one (thio)phosphoric acid triamide is selected from the group consisting of [0138] N-(n-butyl)thiophosphoric acid triamide (NBPT), [0139] N-(n-propyl)thiophosphoric acid triamide (NPPT) [0140] mixtures comprising N-(n-butyl)thiophosphoric acid triamide (NBPT) and N-(n-propyl)thiophosphoric acid triamide (NPPT), and [0141] mixtures comprising N-(n-butyl) thiophosphoric acid triamide (NBPT) and N-(n-propyl) thiophosphoric acid triamide (NPPT) wherein NBPT is contained in amounts of from 50 to 90 wt. % and NPPT is contained in amounts of from 10 to 50 wt. % based on the total amount of active urease inhibitors. [0142] 4. The method according to item 1 or 2, wherein the at least one (thio)phosphoric acid triamide is N-(n-butyl) thiophosphoric acid triamide (NBPT). [0143] 5. The method according to item 1 or 2, wherein the at least one (thio)phosphoric acid triamide is N-(n-propyl) thiophosphoric acid triamide (NPPT). [0144] 6. The method according to item 1 or 2, wherein the at least one (thio)phosphoric acid triamide is a mixture comprising N-(n-butyl) thiophosphoric acid triamide (NBPT) and N-(n-propyl) thiophosphoric acid triamide (NPPT). [0145] 7. The method according to anyone of the items 1 to 6, wherein the plant, or the plant growing on soil or soil substituents and/or the locus where the plant is growing or is intended to grow is treated with at least one (thio)phosphoric acid triamide in amounts of from 0.03 kg/hectare to 0.5 kg/hectare. [0146] 8. The method according to anyone of the items 1 to 7, wherein the crop biomass or crop yield is increased by at least 1%, more preferably at least 2%, most preferably at least 3%, particularly at least 4%, particularly preferably at least 5%, particularly more preferably at least 6%, particularly most preferably at least 7%, for example at least 8%, for example preferably at least 9%, for example more preferably at least 10%, compared to the biomass or yield of the same crop produced under the same conditions, but without the treatment of the invention. [0147] 9. The method according to anyone of the items 1 to 7, wherein improved plant vigor is determined by at least one of the features or parameters selected from the group consisting of improved vitality of the plant, improved plant growth, improved plant development, improved visual appearance, improved plant stand, less plant verse/lodging, improved emergence, enhanced root growth and/or more developed root system, enhanced nodulation, in particular rhizobial nodulation, bigger leaf blade, bigger size, increased plant weight, increased plant height, increased tiller number, increased number of side shoots, increased number of flowers per plant, increased shoot growth, increased root growth such as extensive root system, enhanced photosynthetic activity, preferably based on increased stomatal conductance and/or increased CO2 assimilation rate, increased stomatal conductance, increased CO2 assimilation rate, enhanced pigment content such as chlorophyll content, flowering, earlier fruiting, earlier and improved germination, earlier grain maturity, improved self-defense response, less non-productive tillers, less dead basal leaves, less input needed such as fertilizers or water, greener leaves, complete maturation under shortened vegetation periods, less fertilizers needed, less seeds needed, easier harvesting, ripening, longer shelf-life, longer panicles, delay of senescence, stronger and/or more productive tillers, better extractability of ingredients, improved quality of seeds for being seeded in the following seasons for seed production, altered or reduced production of ethylene and/or the inhibition of its reception by the plant, and growth repression. [0148] 10. The method according to anyone of the items 1 to 7, wherein improved plant quality is determined by at least one of the features or parameters selected from the group consisting of increased nutrient content, increased protein content, increased content of fatty acids, increased metabolite content, increased carotenoid content, increased sugar content, increased content of amino acids, including essential amino acids, improved nutrient composition, improved protein composition, improved composition of fatty acids, improved metabolite composition, improved carotenoid composition, improved sugar composition, improved amino acids composition, improved or optimal fruit color, improved leaf color, higher storage capacity, and higher processability of the harvested products. [0149] 11. The method according to anyone of the items 1 to 7, wherein improved tolerance or resistance of the plant to abiotic stress factors is determined by the improved tolerance and/or resistance to at least one of the stress factors selected from the group consisting of: heat stress including temperatures higher than 30° C., temperature conditions causing heat damage to a plant such as heat damaged foliage or burnt leaves, cold stress such as temperature conditions below 10° C., periods of thawing and freezing, frost, variations in temperature such as temperatures conditions that lead to the freezing of water either for extended periods of time or only temporary periods, temperature unusual for the season, drought stress, exposure to cold water, flood, water-logging, wind, sun light, particularly sun light causing signs of scorch, sun burn or similar signs of irradiation and heat stress to the plant, acid or alkaline pH conditions in the soil with pH values lower than pH 5 and/or pH values higher than 9, salt stress such as soil salinity, soil erosion, inorganic pollution, soil contamination or soil pollution with chemicals, particularly with heavy metals, preferably chromium, lead, cadmium, arsenic, antimony, mercury, iron, thallium, barium, beryllium, polonium, uranium, toxic waste, nuclear waste, acid rain, air pollution, preferably radiation such as high UV radiation due to the exposure to the decreasing ozone layer, increased ozone levels, nitrogen oxides and/or sulfur oxides, oxidative stress, organic pollution, oil and/or fuel dumping or spilling, nuclear radiation, contact with sewage, over-fertilization, nutrient deficiencies, herbicide injuries, plant wounding, compaction, natural disasters, preferably tornadoes, hurricanes, wildfires, flooding and combinations thereof. [0150] 12. The method according to anyone of the items 9, wherein at least one of the features or parameters is improved or increased by at least 1%, more preferably at least 2%, most preferably at least 3%, particularly at least 4%, particularly preferably at least 5%, particularly more preferably at least 6%, particularly most preferably at least 7%, for example at least 8%, for example preferably at least 9%, for example more preferably at least 10%, compared to the plant produced under the same conditions, but without the treatment of the invention. [0151] 13. The method according to anyone of the items 10, wherein at least one of the features or parameters is improved or increased by at least 1%, more preferably at least 2%, most preferably at least 3%, particularly at least 4%, particularly preferably at least 5%, particularly more preferably at least 6%, particularly most preferably at least 7%, for example at least 8%, for example preferably at least 9%, for example more preferably at least 10%, compared to the plant produced under the same conditions, but without the treatment of the invention. [0152] 14. The method according to anyone of the items 11, wherein the tolerance or resistance of the plant to at least one of the stress factors is improved or increased by at least 1%, more preferably at least 2%, most preferably at least 3%, particularly at least 4%, particularly preferably at least 5%, particularly more preferably at least 6%, particularly most preferably at least 7%, for example at least 8%, for example preferably at least 9%, for example more preferably at least 10%, compared to the plant produced under the same conditions, but without the treatment of the invention. [0153] 15. Use of at least one (thio)phosphoric acid triamide according to the general formula (I)

R.sup.1R.sup.2N—P(X)(NH.sub.2).sub.2  (I) [0154] wherein [0155] X is oxygen or sulfur; [0156] R.sup.1 and R.sup.2 are—independent from each other—H, substituted or unsubstituted 2-nitrophenyl, C.sub.1 to C.sub.20 alkyl, C.sub.3 to C.sub.20 cycloalkyl, C.sub.6 to C.sub.20 heterocycloaryl, C.sub.6 to C.sub.20 aryl, or dialkylaminocarbonyl group, wherein R.sup.1 and R.sup.2 together with the nitrogen atom linking them define a 5- or 6-membered saturated or unsaturated heterocyclic radical, which optionally comprises 1 or 2 further heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, [0157] for improving the health of a plant, comprising treating a plant growing on soil or soil substituents and/or the locus where the plant is growing or is intended to grow, wherein improved plant health is determined by [0158] increased crop biomass or crop yield, [0159] improved plant vigor, [0160] improved plant quality, and/or [0161] improved tolerance or resistance of the plant to abiotic stress factors.