RU2513621C2 - Agent exhibiting growth-regulating and immunomodulating activity - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to agriculture and specifically to plant growth regulators. The agent is 5,5-dimethyl-2-(1-(4-methoxyphenyl)-3-phenyl-3-oxopropyl)-cyclohexanedione-1,3

.

EFFECT: along with high growth-regulating activity, the agent has immunomodulating properties.

12 tbl, 3 ex

Description

The invention relates to the field of agriculture, namely to plant growth regulators, and can be used as an immunomodulating agent.

Known means to stimulate growth and development, protection against fungal diseases of crops, which is 7,7-dimethyl-2-phenyl-4- (4-methoxyphenyl) -5-oxo-5,6,7,8-tetrahydroquinoline. The invention can be used to treat seeds or tubers in order to increase their germination and resistance to fungal diseases (see Pat. RF invention No. 2359457, IPC A01N 43/42, A01P 21/00, A01P 3/00).

However, the known agent has insufficient growth-regulating and immunomodulating properties.

Known agent for the treatment and prevention of intestinal helminthiases in animals is a substance of the series 5,5-dimethyl-2- (1-R, 3-phenyl-3-oxopropyl) -cyclohexanedione-1,3, moreover, the substituents (R) may be phenyl or 4-methoxyphenyl groups. The tool has high therapeutic efficacy in ascariasis and other intestinal helminthiases of animals (see US Pat. RF invention No. 2132185, IPC A61K 31/12, A01N 35/06).

However, the known tool was not used to accelerate growth and increase immunity.

The technical result of the proposed solution is to increase the growth-regulating and immunomodulating properties.

The claimed agent exhibiting growth-regulating and immunomodulating activity, which is a compound of 5,5-dimethyl-2- (1- (4-methoxyphenyl) -3-phenyl-3-oxopropyl) cyclohexanedione-1,3

(далее МФК)

(hereinafter IFC)

The claimed agent is obtained by Michael condensation of cyclic (3-diketone-5,5-dimethylcyclohexane-1,3-dione (dimedone) with chalcones benzalacetophenone (R = phenyl) and 4-methoxybenralacetophenone (R = 4-methoxyphenyl) under basic catalysis Piperidine or sodium ethylate is used as a catalyst. The yield of products is from 59 to 87%. The composition and structure of the obtained triketones were confirmed by elemental analysis and IR spectroscopy. Samples of the compounds used in these studies were identified by temperature melting range, mixing samples with previously obtained samples (L.I. Lelyukh, K.M. Korshunova, V. G. Kharchenko. Synthesis of tricetones -1,3-di-aryl-3- (2-dihydroresorcinol) -1- propanones // Chemistry and Chemical Technology. - 1975. - vol. XVIII. - issue 1. - p. 1537-1539.).

Example 1. Laboratory studies of the growth-regulating effect of the IFC preparation to assess the intensity of seedling formation, germination energy and laboratory germination of seeds, peroxidase enzyme activity in seedlings, amylase activity in grain and grain quality.

The experiments were carried out on cucumbers of the Phoenix variety, tomatoes of the Volgograd early variety, spring wheat of the Saratovskaya 68 variety in 2007-2009.

To assess the effect of IFCs on the processes of cucumber and tomato germination, 50 seeds of each culture were soaked for 24 hours in a ^10-3 % IFC solution (10 mg of substance per 1 liter of solution), then washed with water and dried [See Velik V.F., Krotova R.A. Seed quality, methods for their pre-sowing preparation // Sat. Vegetable farming of open ground. - M .: Kolos, 1984. - P.108].

The standard in experiments on cucumbers and tomatoes was a solution of the industrial plant growth stimulator Immunocytophyte (IC), the processing of which was carried out according to the recommended method (see Kulnev A.I. Multipurpose stimulants of the protective reactions of plant growth and development (using the example of the drug Immunocytophyte) / A.I. .Kulnev, E.A. Sokolova. - Pushchino: ONTI PNC RAS, 1997.-100 p.).

The control in the experiments was water.

In studies with spring wheat, the standard was a solution of zenobacterial fertilizer Cenosis, prepared on the basis of a crushed mass of spring wheat straw [See A.S. No. 14548151]. The control was water.

In one embodiment of the experiment on wheat, the soaking of seeds in a MFC solution was combined with the soaking in a solution of Cenose.

In laboratory experiments with wheat, seeds were also grown in the field under different nutritional backgrounds using pre-sowing seed treatment with water and an IFC solution when a complete mineral fertilizer N ₁₆ P ₁₆ K _{16 was} added to the rows before sowing.

To obtain seedlings, the seeds of all cultures were germinated in wet rolls of filter paper in an incubator at 24-25 ° C until germination of 20-40 mm in length [See A large workshop on plant physiology / Gavrilina V.F., Ladygina M.E., Khaydobina L.M. / Higher. school., 1975. - 391 s].

Laboratory germination and seed germination energy was evaluated according to the generally accepted methodology (see GOST 12038-84. Seeds of agricultural crops. Methods for determining germination).

Observations of the formation of seedlings in a Phoenix cucumber and early Volgogradsky tomato varieties showed that the germination energy of both the cucumber and tomato treated with the IFC solution was higher than in the control and than when treated with an industrial stimulant IC (Table 1) . In the IFC variant, both cultures formed more powerful shoots with a well-developed root system.

The results of a study of laboratory germination and energy of germination of wheat seeds indicate that the IFC preparation increases these indicators by 3-6% compared with the control and enhances the effect of the use of Cenosis and N ₁₆ P ₁₆ K ₁₆ (Table 2).

In the seedlings of all studied cultures, the activity of the peroxidase enzyme was determined. The resulting sprouts were separated from the seeds, triturated with cold acetate buffer (pH 4.7) in a ratio of 1:10. The suspension was infused in the refrigerator for 12 hours, then centrifuged at 12,000 g. In the supernatant, the activity of the peroxidase enzyme was determined by the method with orthophenylenediamine (see Suslova T.A., Khorosheva T.M. To the question of biochemical aspects of alfalfa resistance to mycoplasmosis // Sat .: Zap. Rast. - Saratov, 1993. - S.111.).

In all cultures, treatment with the IFC preparation led to a significant activation of the peroxidase enzyme compared to the control and standard (Table 3).

In experiments on wheat, the use of IFC significantly increased the effect of coenosis and N ₁₆ P ₁₆ K ₁₆ . The peroxidase activity in the variants Cenosis + IFC and N ₁₆ P ₁₆ K ₁₆ + IFC increased in comparison with the control by 2475 and 3175 unit units / 100 g of crude material, respectively.

A similar effect from treatment with IFC indicates its high biological activity and ability to be included in redox processes in plants.

The total activity of amylases was determined in wheat grain by the amount of starch not cleaved by amylase on a photocolorimeter after treatment with iodine (see Pleshkov B.P. Workshop on Plant Biochemistry. M .: Vyssh.shk., 1985. - 256 s).

A decrease in the activity of amylases in grain helps to increase the resistance of crops to sprouting, since pre-harvest germination reduces the sowing and technological qualities of grain (see Sergeev A.V. Selection, seed production and cultivation of triticale. - M .: Agropromizdat, 1982. - 30 s )

Under the influence of IFC treatment, amylase activity in wheat grain significantly decreased compared to the control (Table 4).

In the case of the use of MPC in combination with Cenosis and N ₁₆ P ₁₆ K _16, this effect was not observed.

An important indicator of the quality of wheat grain is its content of raw gluten and its quality (IDK).

The determination of the amount of raw gluten in wheat grain and its quality was carried out according to the generally accepted method (see GOST 27839-88 Wheat flour. Methods for determining the quantity and quality of gluten).

When using the IFC preparation, the content of crude gluten in wheat grain was 3-4% higher compared to the control (Table 5).

On average, IFC increased this indicator by 1% when using Cenosis and N ₁₆ P ₁₆ K ₁₆ .

The use of IFCs did not impair the quality of gluten; in all variants of the experiment, the quality of gluten can be attributed to quality group II.

Example 2. Growth-regulating activity of IFC on various crops in open ground.

Field experiments in open ground for the study of IFC were carried out during 2007-2009. Phoenix cucumber in the conditions of the Bazarno-Karabulaksky district of the Saratov region, early Volgogradsky tomatoes in the conditions of the Novo-Burassky district of the Saratov region and spring wheat of the Saratovskaya 68 variety in the conditions of the Arkadaksky district of the Saratov region.

Testing, harvesting and recording yields were carried out according to the method of state variety testing of agricultural crops [See Moscow VASHNIL VIUA. - 1985, issue 2. - 286 s].

Statistical processing of data was carried out according to the Dospekhov method (see B. Dospekhov, Field Experiment Technique. - M.: Agropromizdat, 1985. - 351 s).

Before sowing, the seeds of cucumber and tomatoes were soaked in water, IC and MPA solutions for 24 hours, then washed with water and dried. The consumption rate of the IFC preparation was 50 mg / ha.

Wheat seeds were soaked in water, a solution of Cenosis and a solution of MPA, and in accordance with the experimental plan, they were sequentially treated with Cenosis, then IFC.

In two variants of the experiment on wheat, in the rows before sowing, full mineral fertilizer N ₁₆ P ₁₆ K _{16 was} introduced: for seeds soaked in water and for seeds soaked in an MPA preparation.

The volume of the working solution for soaking 1 ton of seeds was 80 liters.

Tomatoes were planted in open ground in seedlings 45 days after sowing seeds.

Sowing cucumbers and wheat was carried out directly into the soil with treated seeds.

The repetition of the experiments is 4-fold. The size of registration plots for all crops was 25 m ² , the protective zone was 1.5 m.

For accounting, 10 plants were taken from each repetition.

The experiments were laid on chernozem soils. Soil preparation included autumn plowing to a depth of 25-27 cm, spring harrowing, double cultivation. Wheat crops rolled in ring rollers.

Before planting cucumber and tomato seedlings, fertilizers were applied (superphosphate 4 kg / ha, potassium chloride 2 kg / ha), and ammonium nitrate (1.5 kg / ha) was added during cultivation.

During the growing season of cucumber and tomatoes, 6-10 waterings were carried out. Wheat was grown in rainfed conditions.

From table 6 it is seen that under the influence of treatment with IFC, field germination in all crops increased: in cucumber - by 15%, in tomatoes - by 6%, in wheat - by 5% compared with the control. Wheat germination also increased significantly with the combination of the IFC preparation with coenosis and N ₁₆ P ₁₆ K ₁₆ .

The treatment of cucumber and tomato seeds with IFC positively affected the formation of earlier commercial products. The stages of the onset of organogenesis under the action of IFCs moved to an earlier date (Table 7).

From tables 8 and 9 it follows that the pre-sowing treatment of the seed material of cucumbers and tomatoes with a solution of IFC contributed to an increase in yield by 11% and 51%, respectively.

Moreover, in both cultures, in the variant with IFC, the number of fruits from one plant increased significantly, the fruits formed larger.

Seed treatment with an IFC for spring wheat cultivation changed positively such morphological indicators as plant height and spike length (Table 10).

In all variants of experiments on wheat using IFCs, the indicators of productivity elements increased (Table 11).

When treating wheat seeds with IFC solution, the number of spikelets per spike increased by 29% compared to the control, the amount of grain in the spike - by 15.5%, the weight of 1000 grains - by 10.4%, the yield - by 14.1%.

The combination of IFCs with Cenosis and complete mineral fertilizer contributed to an even greater increase in productivity - by 23.4 and 32.5%, respectively.

The data presented indicate their own growth-regulating effect of the IFC preparation on vegetables and grain crops, as well as its ability to intensify the effect of environmentally friendly zoobacterial fertilizer (Cenosis).

Example 3. The study of IFC as an inducer of resistance to fungal diseases in vegetable crops (see. Protection of plants from diseases // edited by V.A. Shkalikov - M .: Kolos, 2003. - P.254.).

In the field, cucumbers and tomatoes took into account the degree of development of the main fungal diseases: on cucumber, powdery mildew; on tomatoes, the prevalence and prevalence of late blight and the prevalence of alternariosis (see Phytosanitary examination of crops (recommendations). M: Rosinformagroteh of the Ministry of Agriculture, 2002.-134 p.).

Accounting for tomato diseases was carried out by examining the plants in the vine and during the collection of fruits (see Orekhovskaya MV, Gurkina LK Diseases of tomatoes // Plant Protection. - 1990. - No. 3. - P.54).

The account of the degree of development of powdery mildew on a cucumber was carried out in three stages: the beginning of flowering, the first fruiting, mass fruiting (see Methodology VIZR, 1985).

The results of field trials indicate that IFC intensifies the processes of plant cell metabolism and induces their resistance to diseases.

Under the influence of IFC, the prevalence of powdery mildew of cucumber at the stage of mass fruiting was 4% less than in the control (Table 12).

The inducing effect of IFC was also detected on tomatoes (Table 12). Under the influence of the treatment, blight decreased by 50%, the prevalence of late blight decreased by 40%, to a lesser extent in the variant with IFC, alternariosis was manifested in tomatoes.

The results of field experiments to study the inducing effect of IFCs directly correlate with laboratory data to determine the activity of the peroxidase enzyme (see Example 1).

In a plant cell, peroxidase performs a protective function, therefore, the enhancement of peroxidase activity in plants can be considered one of the forms of their biological protection (see Andreeva V.A. Enzyme peroxidase. - M .: Nauka, 1988. - 125 p.).

Under the influence of IFC, the activity of peroxidase in seedlings of cucumber, tomato and wheat increased significantly (Table 3).

It can be assumed that under the action of MPC in the plant cell, oxidation - reduction processes are intensified and, as a result, hydrogen peroxide accumulates, which is one of the active forms of oxygen that takes part in inducing plant immunity (see Ozerskovskaya OL, Vasyukova N. I. The effect of immunomodulators on the resistance and susceptibility of potatoes to Phytophtora infestans // Plant Physiology. - 2006. - T.53, No. 4. - S.546-553).

Thus, as a result of laboratory experiments and field trials, a high growth-regulating and immunomodulating activity of the IFC preparation was established.

The IFC preparation intensifies the processes of cell metabolism, including redox, which leads to positive morphological changes in plants; the formation of a more powerful root system and aerial parts, the intensification of the stages of vegetation, the development of generative organs.

Under the influence of IFCs, there is an increase in total marketable yield, an increase in the resistance of plants to diseases, which leads to an increase in the level of profitability of production.

So the recoupment of costs on a cucumber when applying IFC was 700 rubles. against 300 rubles. in the version with IC, on tomatoes - 80.7 rubles. (IFC) versus 64.8 rubles. (IC), on wheat the level of profitability in options with IFC varied from 102.5 to 121.4%.

Table 1.

The effect of IFC on the formation of seedlings of cucumber of the Phoenix variety and tomatoes of the Volgograd early variety (average for 2007 - 2009)

Indicator Cucumbers Tomatoes Control, H ₂ O Standard, IC IFC, 10 ^-3 % Control, H ₂ O Standard, IC IFC, 10 ^-3 % Sprout length, cm 4.2 5.8 6.4 1,2 1,6 2.1 Root length, cm 3.0 5.8 7.9 1.8 2.1 2,8 Germination energy,% 52 58 62 48 51 56

Table 2.

The influence of IFCs and methods of presowing treatment of seeds of spring wheat Saratovskaya 68 on their sowing qualities (average for 2007-2009)

Indicators
Options Germination energy,% Laboratory germination,% The mass of seedlings, g Control, N ₂ O 52 88 5,4 Standard, Cenosis 64 92 7.0 IFC, 10 ^-3 % 60 91 6.0 Cenosis + IFC · 10 ^-3 % 66 94 7.6 N ₁₆ P ₁₆ K ₁₆ 67 93 7.0 N ₁₆ P ₁₆ K ₁₆ + IFC · 10 ^-3 % 67 95 7.6 Hcp ₀₅ 2.2 1,6 0.4

Table 3

The effect of IFCs and presowing treatment of seeds on peroxidase activity (specific units / 100 g of raw material) in different crops (average for 2007-2009)

Culture Options Peroxidase activity Cucumber Control, N ₂ O 59 Standard, IC 101 IFC, 10 ^-3 % 150 Tomatoes Control, H ₂ O 28 Standard, IC 42 IFC, 10 ^-3 % 58 Wheat Control, H ₂ O 3321 Standard, Cenosis 4552 IFC, 10 ^-3 % 5334 Cenosis + IFC · 10 ^-3 % 5796 N ₁₆ P ₁₆ K ₁₆ 4934 N ₁₆ P ₁₆ K ₁₆ + IFC · 10 ^-3 % 6496

Table 4

The effect of IFCs and agricultural practices on the activity of amylases in grain of spring wheat Saratovskaya 68 (average for 2007-2009)

Table 5

Influence of IFCs and agricultural practices on grain quality of spring wheat Saratovskaya 68 (average for 2007-2009)

Indicators Experience Options Control, H ₂ O Standard, Cenosis IFC, 10 ^-3 % Cenosis + IFC · 10 ^-3 % N ₁₆ P ₁₆ K ₁₆ N ₁₆ P ₁₆ K ₁₆ + IFC · 10 ^-3 % Crude gluten,% 25.1 27,2 27.5 28,4 27.6 28.8 Idk 95 80 76 80 76 77

Table 6

The effect of the IFC preparation and agricultural practices on the field germination of cucumber, tomato, wheat (average for 2007-2009)

Culture Options Field germination,% Cucumber Control, N ₂ O 60.0 Standard, IC 71.0 IFC, 10 ^-3 % 75.0 Tomatoes Control, N ₂ O 81.0 Standard, IC 83.0 IFC 10 ^-3 % 87.0 Wheat Control, N ₂ O 76.3 Standard, Cenosis 80.3 IFC, 10 ^-3 % 81.3 Cenosis + IFC · 10 ^-3 % 84.1 N ₁₆ P ₁₆ K ₁₆ 82.7 N ₁₆ P ₁₆ K ₁₆ + IFC · 10 ^-3 % 87.2

Table 7

The effect of IFC on the passage of the stages of organogenesis in cucumber and tomatoes in the open ground (average for 2007-2009)

Experience Options Cucumber Tomatoes Bloom Beginning of fruiting Bloom Beginning of fruiting Mass bearing Maturation Control, H ₂ O 07/26 08/08 2906 07/15 07/22 07/30 Standard, IC 07/26 07/29 06/27 12.07 07/21 07/26 IFC, 10 ^-3 % 07/25 07/28 06/25 11.07 07/18 07/25

Table 8

Influence of IFC on the productivity of Phoenix cucumber (average for 2007-2009)

Experience Options The number of fruits from 1 plant, pcs. The average weight of 1 fetus, g Harvest, kg / m ² % Increase to control Control, N ₂ O 18.0 136.5 25.0 - Standard, IC 21.3 150.0 28.1 7.7 IFC, 10 ^-3 % 22.8 150.0 29.1 11.5 NDS ₀₅ 0.4 4.1 0.7

Table 9

The influence of IFC on the productivity of tomato of the early Volgograd variety (average for 2006-2009)

Experience Options The number of fruits from 1 plant, pcs. Weight 1 fruit, g Productivity, kg / m ² % Increase to control Control, H ₂ O 25 80 10,4 - Standard, IC 27 one hundred 13.5 17 IFC, 10 ^-3 % 29th 120 17.4 51 NDS ₀₅ 0.89 13.0 1.7

Table 10

The influence of IFCs and agricultural practices on the morphological parameters of wheat Saratovskaya 68 (average for 2007-2009)

Options
Indicators Control, H ₂ O IFC, 10 ^-3 % Cenosis Cenosis + IFC · 10 ^-3 % N ₁₆ P ₁₆ K ₁₆ N ₁₆ P ₁₆ K ₁₆ + IFC · 10 ^-3 % Plant height, cm 68.3 75,4 70,2 76,4 77,2 78.7 Spike length, cm 6.3 7.5 6.8 7.4 7.4 7.6

Table 11

The effect of IFCs and agricultural practices on wheat productivity Saratovskaya 68 (average for 2007-2009)

Indicators
Options The number of spikelets in one spike, pcs. The amount of grain in the ear, pcs. Mass of 1000 grains, g Productivity t / ha % Increase Control, N ₂ O 9,4 19.3 33,4 1.51 Standard, Cenosis 11,4 22.0 36.3 1,66 9.9 IFC, 10 ^-3 % 12.1 22.3 36.9 1.72 14.1 Cenosis + IFC · 10 ^-3 % 12.6 23.7 37.8 1.86 23,4 N ₁₆ P ₁₆ K ₁₆ 10,4 23.7 37.7 1.81 19.9 N ₁₆ P ₁₆ K ₁₆ + IFC · 10 ^-3 % 14.0 25.3 39.5 2.00 32,5 NDS ₀₅ 0.40 0.72 1.12 0.11

Table 12

The effect of IFC on the prevalence of cucumber and tomato by major diseases (average for 2007-2009)

Options Cucumber, powdery mildew,% Tomatoes Beginning of flowering Beginning of fruiting Mass bearing Late blight Alternariosis, affection, score Prevalence,% Spread.% Control, H ₂ O 3.75 6.25 10.0 70 70 four IC 0 3.15 7.25 45 60 four IFC, 10 ^-3 % 0 2.15 6.25 twenty thirty 3

Claims (1)

The agent exhibiting growth-regulating and immunomodulating activity, which is a compound of 5,5-dimethyl-2- (1- (4-methoxyphenyl) -3-phenyl-3-oxopropyl) cyclohexanedione-1,3