RU2587553C2 - Method of increasing number of root nodules of plant - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to use of sedaxan (SDX) for increasing the number of tubercules on plant roots compared with the number of tubercules on roots raw control plants by using the above SDX to a plant, and/or its location, and/or material for propagation of plants, and then growing said plants or material for propagation. In a specific embodiment, the above plant is a soya and said material for propagation represents seeds.

EFFECT: invention enables to realise the specified purpose.

15 cl, 3 tbl, 1 ex

Description

The present invention relates to the use of sedaxan to produce a plant having an increased number of nodules on its roots. In particular, the invention relates to the use of sedaxan to produce a soybean plant having a larger number of nodules.

The binding of atmospheric nitrogen (N ₂ ) to form nitrogen-containing organic compounds is called nitrogen fixation. In plants, symbiotic nitrogen binding occurs in nodules, usually legumes, of plants, whereby bacteria called Rhizobia, which otherwise live freely in the soil, infect roots and colonize bark cells. Bacteria utilize carbohydrates from the plant's phloem, in exchange providing the plant with nitrogen-containing products through xylem. Rhizobia belongs to the families Rhizobiales and Burkholderiales, the most significant species belong to the genera Rhizobium and Bradyrhizobium, but many other species in some other genera Rhizobiales or Burkholderiales have been noted in their ability to fix atmospheric nitrogen.

Plants that can efficiently apply the nitrogen fixation process are desirable from the point of view of commercial farming because they can reduce the amount of additional fertilizer needed to get an acceptable crop.

With the growing world population, there is still a need to create even more effective ways to maximize returns on increasingly valuable and expensive world agricultural lands.

It has been unexpectedly discovered that fungicide sedaxan can increase the number of nodules on plant roots, thus leading to an increase in the possibility of colonization with a nitrogen-fixing bacterium such as Rhizobium or Bradyrhizobium, leading to an increase in the availability of nitrogen in the plant itself.

The present invention provides the use of sedaxan (SDX) to increase the number of nodules on the roots of a plant by applying said SDX to the plant and / or its location and / or material for propagating the plant, and then growing said plant or material for propagating the plant where the number of nodules on the roots of said plant is higher than the number of nodules on the roots of the untreated control plant.

Sedaxan is a pyrazolecarboxamide: 3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxylic acid fungicide based on (2-bicyclopropyl-2-yl-phenyl) -amide having the formula:

He and his trans- and cis-isomeric forms, namely:

also described in various published international patent applications, including: WO 03/074491, WO 2006/015865, WO 2007/090623, WO 2007/131678, WO 2008/000377, WO 2008/003403, WO 2009/098218, WO 2010/000790 and disclosed research information [(2006), 507 (July), P783-P791 (Number 507002) CODEN: RSDSBB; ISSN: 0374-4353].

An additional preferred embodiment of the present invention is represented by the use of sedaxan or a method in which sedaxan is used to increase the number of nodules on the roots of a plant, where sedaxan is present in a composition that includes a racemic mixture of trans isomers and cis isomers of sedaxan with respect to trans / cis from 1: 1 to 100: 1, for example 1: 1, 2: 1, 3: 1, 4: 1, 5: 1, 6: 1, 7: 1, 8: 1, 9: 1, 10: 1, 20: 1 , 50: 1 or 100: 1. Further preference is given to ratios from 2: 1 to 100: 1, more preferably from 4: 1 to 10: 1. In a further embodiment, sedaxan is applied to the plant and / or its location and / or material for propagating the plant in a composition in which sedaxan is present in an amount of 65-99% of its trans isomer.

The present invention further provides the use of a compound according to formula (I):

Where

R ₁ represents trifluoromethyl or difluoromethyl and

R ₂ represents hydrogen or methyl; or a tautomer of such a compound, to increase the number of nodules on the roots of the plant by applying said compound to the plant and / or its location, and / or material for propagating the plant, and then growing said plant or propagation material, where the number of nodules on the roots of said plant higher than the number of nodules on the roots of the untreated control plant.

The present invention provides a method for producing a plant with an increased number of nodules on the roots of said plant, said method comprising applying sedaxan to said plant and / or its location and / or plant propagation material, and then growing said plant or plant propagation material where the number of nodules on the roots of said plant is higher than the number of nodules on the roots of the untreated control plant.

When used, any composition containing sedaxan may also include additional active ingredients, for example, an insecticide, fungicide, nematicide, synergist, herbicide, plant growth regulator or a compound that promotes "plant health". Examples of active ingredients that can be added to a sedaxan-containing composition include the compounds listed in The Pesticide Manual (British Crop Production Council - ISBN No. 9781901396188), which can be found at www.bcpc.org. Particularly preferred mixtures include those disclosed in WO 2006/015865 and in WO 2007/090623.

If necessary, the use of sedaxan and an additional active ingredient (for example, in the form of a “combination”) can be carried out in the form of a single “finished mixture” such as a finished composition comprising two components in the prescribed ratio; or in a combined spraying mixture or a mixture for use composed of separate formulations of individually taken active ingredients as components (for example, “tank mixture”); or by combined use of the individual active ingredients in a general plan or spray schedule in a consistent manner, i.e. one after another for an acceptable short period (for example, with an interval of several hours one from another). In the case of application in separate sprayings or in seed treatments, following one after another, the order of application of the active ingredients is not so important to achieve the effect in accordance with the present invention.

The present invention also further provides a use or method as described above, wherein said plant or plant propagation material is soy.

The present invention also further provides a use or method as described above, wherein said plant propagation material is seeds.

The present invention also further provides a use or method as described above, where sedaxane is applied to said plant and / or its location and / or plant propagation material in such an amount that the number of nodules on the roots of the plant is at least 5% higher than the number of nodules on the roots of the untreated control plant.

The present invention also further provides a use or method as described above, where sedaxane is applied to said plant and / or its location and / or plant propagation material in such an amount that the number of nodules on the roots of the plant is at least 10% higher than the number of nodules on the roots of the untreated control plant.

The present invention also further provides a use or method as described above, where sedaxane is applied to said plant and / or its location and / or plant propagation material in such an amount that the number of nodules on the roots of the plant is at least 50% higher than the number of nodules on the roots of the untreated control plant.

The present invention also further provides a use or method as described above, where sedaxane is applied to said plant and / or its location and / or plant propagation material in such an amount that the number of nodules on the roots of the plant is at least 60% higher than the number of nodules on the roots of the untreated control plant.

The present invention also further provides a use or method as described above, wherein said plant grows in the presence of Rhizobium sp. and / or Bradyrhizobium sp.

The present invention also further provides a use or method as described above, wherein the plant contains a higher percentage of nitrogen in its foliage compared to the untreated control plant.

The present invention also further provides use as described above, wherein said plant grows in the presence of Rhizoctonia sp.

The present invention also further provides use as described above, wherein said plant grows in the absence of Rhizoctonia sp.

Rhizoctonia is a genus of anamorphic fungi in the order Cantharellales. Varieties of Rhizoctonia are plant pathogens that cause commercially important plant diseases, in particular root diseases.

"Plant" and "plants" in accordance with this invention encompass plants that are capable of forming nodules on roots that can be colonized by strains of Rhizobium sp. or Bradyrhizobium sp. Typically, such plants include legumes of the Fabaceae family. Examples of legumes include beans, lentils, peas, soybeans, castor oil plants, cocoa beans or peanuts. Examples of legumes commonly used for animal consumption include: alfalfa; asparagus beans; asparagus peas; small lima beans; common dolichos; cow peas; black tortoise bean; Yankee Beans; white navy beans; fodder beans; Cannellini beans chickpeas; chili beans; cacao beans; Cranberry Beans; dwarf beans; hyacinth beans; Egyptian lobia; English beans; fava beans; field peas; french green beans; great northern bean; green beans; green and yellow peas; Kidney Beans; lentils; lespedetsu; licorice; lima beans; Madagascar Beans; Mexican black beans; Mexican Red Beans; yellow eye beans; golden beans; golden peas; navy beans; fine seed beans; peanut; Peruvian Beans; pinto beans; red bean; Red clover; red eye beans; red Kidney beans; rice beans; green beans; red string beans; small red beans; small white beans; peas; southern peas; sugar peas; soy; bean beans; white clover; White Kidney Beans and White Small Seed Beans. This list is not a limitation.

The terms “plant” and “plants” also include plants that have been given resistance to herbicides, insecticides, fungicides or have been modified in any other way, for example, to increase yield, drought tolerance or quality, using traditional breeding methods or by genetic engineering methods . Any genetically modified plants used according to the present invention could be modified by applying recombinant nucleic acid techniques well known to those skilled in the art. For example, “plant” and “plants” according to the present invention include those soy plants that have been bred by selection methods so that they are resistant to Asian soybean rust. Such soybean plant varieties are designated by the term “Inox” varieties and include varieties registered in Brazil under the Plant Variety Protection Act under the names TMG 801 and TMG 803.

The expression “location” of beneficial plants, as used herein, is intended to encompass a place where beneficial plants are grown, where propagation material for beneficial plants is sown, or where material for propagation of beneficial plants will be placed in the soil. An example of such a location is the field on which cultivated plants are grown.

The expression “plant propagation material” should be understood as meaning the generative organs of a plant, such as seeds that can be used to propagate the latter, and vegetative material, such as cuttings or tubers. This term includes seeds (in the strict sense), roots, fruits, tubers, bulbs, rhizomes or parts of plants. You can also note the sprouted plants and seedlings that need to be transplanted after germination or after emergence from the soil. These seedlings can be protected before transplantation by full or partial processing by immersion. Preferably, “plant propagation material” is to be understood as meaning seeds.

The term "untreated control plant" means a plant, its location or material for propagation of a plant, which is in almost identical conditions with a plant, the location of which or material for propagation of a plant according to this invention, with the exception of an untreated control plant (or its location or propagation material plants) were not treated with sedaxan. A person skilled in the art is familiar with properly controlled experiments and thus can conduct a comparative study by growing two groups of plants of the same variety / variety under the same conditions, where only one group (representing the plant, its location or plant propagation material) is exposed treatment with sedaxan, thereby allowing the specified specialist to establish the effect of treatment of plants with sedaxane on the formation of nodules by comparison with untreated controls.

The terms “root” and “roots” should be considered to mean the organ of a plant, which usually lies below the surface of a growth medium, such as soil. So, in the context of determining the number of nodules, they are counted by determining the number of nodules in the whole mass of what is usually considered the root of the plant.

The term "sedaxan" may be replaced by the term "SDX".

In yet a further aspect, the present invention also provides a use or method, as described above, to further improve plant health.

For example, useful properties that can be additionally obtained in combination with an increased number of root nodules, as described above, include: improved crop characteristics, including: germination, crop yield, protein content, improved plant power, faster / delayed ripening, increased seed germination rate, improved nutrient absorption efficiency, improved nitrogen absorption efficiency, improved water use efficiency, improved content oil quality and / or quality, improved digestion, faster / more uniform ripening, improved taste, improved starch content, more developed root system (improved root growth), improved stress tolerance (e.g. against drought, heat, salt, light, UV , water, cold), reduced ethylene (decreased formation and / or inhibition of perception), increased shoot formation, increased plant height, a larger leaf blade, less withering of the main leaves, stronger shoots, more green leaf color, pigment content, photosynthesis activity, less required amount of application (e.g. fertilizer or water), less needed seeds, more fruitful shoots, earlier flowering, early ripening of seeds, less lodging of plants (lodging), increased seedling growth, increased plant power, increased plant resistance to lodging and early germination.

The beneficial properties obtained, especially from the treated seeds, include, for example, improved germination and rooting in the field, better plant power and more uniform survival in the field.

The beneficial properties obtained, especially when applied to leaves and / or furrows, include, for example, improved plant growth and plant development, better growth, more shoots, greener leaves, larger leaves, greater biomass, better roots, improved plant tolerance to stress , larger grain yield, larger biomass collected, improved crop quality (content of fatty acids, metabolites, oils, etc.), more easily sold products (e.g., improved size), improved process (e.g., longer shelf life bottoms, easier extraction of substances), improved seed quality (for sowing in the next season for seed production); or any other advantages familiar to a person skilled in the art.

In a specific embodiment of the invention, as described above, there is provided a plant having a greater number of nodules on its roots than the roots of a control plant, which plant also exhibits improved plant health in addition to a higher number of nodules on its roots.

When applied to a plant in accordance with the use / method of this invention, sedaxan (active ingredient - ai) is usually applied at application rates of 5 to 200 g ai / ha, in particular 10 to 1000 g ai ./ha, for example 50, 75, 100 or 200 g ai / ha, usually in combination with from 0.5 to 1000 g / ha, preferably from 1 to 750 g / ha, more preferably from 2.5 to 500 g / ha, more preferably 5 to 300 g / ha, more preferably 7.5 to 200 g / ha of the active ingredient.

In agricultural practice, the application rates of the compositions according to the use / method of the present invention depend on the type of effect required and, as a rule, range from 20 to 4000 g of the whole combination per hectare.

When using sedaxan for seed treatment, application rates of from 0.001 to 50 g sedaksan per kg of seeds, preferably from 0.01 to 10 g per kg of seeds, are usually sufficient.

A composition containing sedaxan and any mixtures with other compounds / active ingredients, as described above, can be used in any conventional form, for example in the form of a double package, powder for dry seed treatment (DS), emulsion for seed treatment (ES), fluid concentrate for seed treatment (FS), seed treatment solution (LS), water-dispersible seed treatment powder (WS), seed treatment capsule suspension (CF), seed treatment gel (GF), emulsion concentrate (EC), concentrate suspensions (SC), suspoemulsions (SE), capsular suspensions (CS) of water-dispersible granules (WG), emulsifiable granules (EG), water-in-oil emulsions (EO), oil-in-water emulsions (EW), microemulsions (ME), oil dispersion (OD) miscible with fluid concentrate oil (OF), miscible with fluid oil (OL), soluble concentrate (SL), suspension for ultra-low-volume application (SU), liquid for ultra-low-volume application (UL), technical concentrate (TC), dispersible concentrate (DC), wettable powder (WP) or any technically feasible composition in combination with acceptable in villages farm economy excipients.

Such mixtures and compositions can be prepared by a method well known to those skilled in the art, for example by mixing the active ingredients with at least one suitable inert excipient for the formulations, for example, diluents, solvents, fillers and optionally other formulation ingredients such as surfactants, biocides, antifreezes, adhesives, thickeners and compounds that give additional effects. One of skill in the art should carefully select inactive ingredients, especially biocides, so that they do not inhibit or otherwise harm nitrogen-fixing bacteria. If a sustained action is desired, then conventional slow release formulations may also be used. In particular, formulations that are applied in atomized states, such as water dispersible concentrates (e.g. EC, SC, DC, OD, SE, EW, EO, etc.), wettable powders and granules, may contain surfactants substances such as wetting and dispersing agents and other compounds that give additional effects, for example, a condensation product of formaldehyde with naphthalene sulfonate, alkylaryl sulfonate, lignin sulfonate, fatty alkyl sulfate and ethoxylated alkyl phenol, and ethoxylated fatty alcohol.

The constituent ingredients well known to those skilled in the art may, for example, include those constituent ingredients that do not have any significant biological activity or do not have biological activity at all. These include, for example, diluents, solvents, fillers, surfactants, biocides, antifreezes, adhesion promoters, thickeners and auxiliary agents.

The composition for seed dressing is used by a method known to a person skilled in the art, for example, in the form of an aqueous suspension or dry powder, having good adhesion to the seeds. Such seed dressing compositions are known in the art. Seed dressing compositions may contain the individual active ingredients or a combination of the active ingredients in an encapsulated form, for example, as slow release capsules or microcapsules. A typical composition of the tank mixture for seed treatment contains from 0.25 to 80%, especially from 1 to 75%, of the required ingredients, and from 99.75 to 20%, especially from 99 to 25%, of solid or liquid excipients (including, for example, a solvent such as water), where the auxiliary substances can be surfactants in an amount of from 0 to 40%, especially from 0.5 to 30%, based on the composition of the tank mixture. A typical composition of the seed treatment mixture contains from 0.5 to 99.9%, especially from 1 to 95%, of the required ingredients, and from 99.5 to 0.1%, especially from 99 to 5%, of a solid or liquid auxiliary substances (including, for example, a solvent such as water), where the auxiliary substances can be surfactants in an amount of from 0 to 50%, especially from 0.5 to 40%, based on the composition of the finished mixture.

Typically, the compositions include from 0.01 to 90% by weight of the active agent, from 0 to 20% of an agriculturally acceptable surfactant and from 10 to 99.99% of solid or liquid inert substances and excipients (substances), and the active agent contains at least sedaxan and optionally other active agents, including the above and / or microbiocides or preservatives, or the like. Concentrated forms of the compositions typically contain from about 2 to 80%, preferably from about 5 to 70% (wt.) Of the active agent. Formulations for application may, for example, contain from 0.01 to 20% (wt.), Preferably from 0.01 to 5% (wt.) Of the active agent. Since commercial products are predominantly in the form of concentrates, the consumer usually uses diluted formulations.

The following example serves to illustrate the invention. The present invention is not limited to this Example.

Example

The next experiment was carried out in a laboratory and in a greenhouse. The strain Rhizoctonia used in this experiment was obtained from the city of Durados, Mato Grosso do Sul. The strain used was Solo 44.

After obtaining the strains, they were purified and grown in a nutrient medium based on potato-dextrose agar (PDA). In conclusion, they were grown in a chamber with controlled temperature (22 +/- 1 ° C) and controlled daylight hours (12 hours in the light / 12 hours in the dark) for 7 days.

The investigated treatments in the experiment were carried out in accordance with Table 1.

Table 1 Number p / p Inoculation Chemical treatment Chemical application rate (ml of product / 100 kg of seeds) one - - - 2 - sedaxan 200 3 Rhizobia - No Rhizoctonia - Yes - - four Rhizobia - No Rhizoctonia - Yes sedaxan 200 5 Rhizobia - Yes Rhizoctonia - Yes - - 6 Rhizobia - Yes Rhizoctonia - Yes sedaxan 200 7 Rhizobia - yes Rhizoctonia - no - - 8 Rhizobia - yes Rhizoctonia - no sedaxan 200

The formulation used was FS (flowing seed concentrate) containing 500 g / L sedaxane.

Estimated parameters:

- Fresh mass of the airborne part of the plant (g / plant);

- Fresh root mass (g / plant);

- The number of nodules;

- Dry mass of nodules;

- The total nitrogen content in the leaves.

Bradyrhizobium infectivity rating:

The Roos soybean plant was grown in a greenhouse after its seeds were treated with sedaxan according to the list of treatments. Immediately after treatment with a chemical (following the instructions of the Ministry of Agriculture), the seeds were inoculated with a Cell Tech® commercial culture containing SEMIA 5079 and SEMIA 5080 Bradyrhizobium japonicum strain cultures, also according to the list of treatments.

Plants were grown in modified Leonard pots (Vincent, 1970 JM Vincent, A manual for the practical study of root-nodule bacteria, IBP Handb. No. 15, Blackwell, Oxford (1970), p.164.) Filled with planting medium, consisting of a mixture of vermiculite, washed sand and calcined charcoal in a ratio of 3: 1: 1, respectively. The pots were lightly watered, covered with rough brown paper and sterilized at 121 ° C and a pressure of 1 atm for 1.5 hours.

After cooling the pots in them, on top of a certain amount of growth medium, mugs of 2.5 cm diameter PDA medium were laid out, infected with a strain of Rhizoctonia, according to the list of treatments. Then the pots were completely filled with growth medium (approximately another 3 cm), then the seeds were sown so that they were not in direct contact with the Rhizoctonia strain.

Pots were regularly watered with McKnight's nutrient-free nitrogen-free solution, alternating with distilled water.

After germination, sprouted and dead sprouts were counted. Excess shoots were then removed, leaving one shoot in each pot. Sprouts were monitored every day, determining their development and symptoms of any diseases.

Bradyrhizobium Nitrogen Fixation Efficiency

During flowering, soybean plants were harvested, the airborne part was separated from the roots and dried in a heating cabinet at 65 ° C to constant weight. The dry weight of this fraction was determined.

Then the dry mass was burned and the total amount of nitrogen was determined using the method of mineralization and titration (microanalysis using the Kjeldahl method). Jackson, 1964 M.L. Jackson, Soil Chemical Analysis, Prentice Hall Inc. Englewood Cliffs, New York (1964) pp. 86-92.

The roots were washed with water to remove soil, then the nodules were separated. The roots were dried at 65 ° C to constant weight. The dry weight of this fraction was determined. Nodules were counted and then dried in a heating cabinet at 65 ° C to constant weight. The dry weight of this fraction was determined.

Statistical analysis

The tests were carried out according to the type of randomized blocks (8 treatments and 5 repetitions). Analyzed data discrepancies. In test F, the significance of differences was 5%, and the average values for the treatments were compared with the Tukey test at 5%.

The results are shown in tables 2 and 3.

table 2 The effect of treatments on the number and dry weight of nodules Number p / p Number of nodules Dry weight of nodules (mg / g) one 21.8 b 0.124 b 2 24.6 b 0.13 b 3 15.4 b 0,1 b four 18.2 b 0,098 b 5 75.5 a 0.434 a 6 80 a 0.408 a 7 72 a 0.386 a 8 113 a 0.524 a

The average values with the same letter designations after them do not differ from the Tukey test (5% confidence).

Table 3 The effect of treatments on the nitrogen content, dry mass of airborne parts and roots Number p / p Nitrogen (%) Dry mass of airborne part (g / plant) Dry mass of roots (g / plant) one 1,984 b 1,598 b 0.906 b 2 2,866 b 1,640 b 1,022b 3 2,576 b 1,236 b 0.806 b four 2,562 b 1,226 b 0.862 b 5 3,376 ab 3,550 a 1,868 a 6 4,020 a 4,158 a 1,800 a 7 3,266 ab 3,304 a 1,546 ab 8 3,980 a 4,406 a 1,818 a

The average values with the same letter designations after them do not differ from the Tukey test (5% confidence).

Claims (15)

1. The use of sedaxan (SDX) to increase the number of nodules on the roots of the plant by applying the specified SDX to the plant, and / or its location, and / or material for propagating the plant, and then growing the specified plant or propagation material, where the number of nodules on the roots said plant is higher than the number of nodules on the roots of the untreated control plant. 2. Use according to claim 1, wherein said plant or propagation material is soy. 3. Use according to claim 1 or 2, wherein said propagation material is seeds. 4. The use according to claim 1 or 2, where the specified SDX is applied to the specified plant, and / or its location, and / or material for propagation of the plant in such an amount that the number of nodules on the roots of the plant is at least 5% higher, than the number of nodules on the roots of the untreated control plant. 5. The use according to claim 4, where the specified SDX is applied to the specified plant, and / or its location, and / or material for propagation of the plant in such an amount that the number of nodules on the roots of the plant is at least 10% higher than the number nodules on the roots of an untreated control plant. 6. The use according to claim 4, where the specified SDX is applied to the specified plant, and / or its location, and / or material for propagation of the plant in such an amount that the number of nodules on the roots of the plant is at least 50% higher than the number nodules on the roots of an untreated control plant. 7. The use according to claim 4, where the specified SDX is applied to the specified plant, and / or its location, and / or material for propagation of the plant in such an amount that the number of nodules on the roots of the plant is at least 60% higher than the number nodules on the roots of an untreated control plant. 8. Use according to claim 1 or 2, wherein said plant grows in the presence of Rhizobium sp. and / or Bradyrhizobium sp. 9. Use according to claim 8, where the plant contains a higher percentage of nitrogen in its foliage compared to the untreated control plant. 10. Use according to claim 1 or 2, wherein said plant grows in the presence of Rhizoctonia sp. 11. Use according to claim 1 or 2, wherein said plant grows in the absence of Rhizoctonia sp. 12. A method of producing a plant with an increased number of nodules on the roots of said plant, said method comprising applying sedaxan to said plant and / or its location and / or plant propagation material, and then growing said plant or plant propagation material, where the number of nodules on the roots of the specified plant is higher than the number of nodules on the roots of the untreated control plant. 13. The method according to claim 12, wherein said plant or propagation material is soy. 14. The method according to item 12 or 13, where the specified material for propagation is a seed. 15. The method according to item 12 or 13, where the specified SDX is applied to the specified plant, and / or its location, and / or material for propagation of the plant in such an amount that the number of nodules on the roots of the plant is at least 5% higher, than the number of nodules on the roots of the untreated control plant.