RU2257059C1 - Growth-controlling complex, method for production thereof, preparation containing the same and uses in agriculture - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: air-dried raw material from terraneous parts of Echinacea purpurea plants is extracted with 90-95 % ethanol in ethanol/raw material ratio of 59-61:1, respectively. Extract is acidified with acid to obtain pH not more than 5.0. Extract is purified with polymeric sorbent and concentrated by vaporization. Obtained plant growth controlling agent contains chicory, chlorogenic and caffeic acids. Growth-controlling preparation contains growth-controlling complex in amount of 0.009-0.11 mg/ml and oxyethylated alkylphenol surfactant in ratio of 1:1. Claimed complex is useful in plant and scrub treatment and in seed soak.

EFFECT: preparation useful as immunomodulator and antistress agent.

6 cl, 6 ex

Description

The present invention relates to plant growth regulators used in crop production to improve the quality of seeds by enhancing germination and enhancing germination energy, enhancing adaptive properties and increasing crop yields, stimulating root formation processes in plants, including the decorative direction, for flowering induction, as well as increase resistance to phytopathogens.

Plant growth regulators are chemicals that affect plant growth and development. Used to accelerate ripening, increase productivity and quality of crops; increase their frost, drought and salt tolerance; fight against lodging of crops, flax and with drawing out vegetable seedlings; suppressing the growth of young shoots and the development of axillary buds in order to increase the fruiting intensity of some fruit and berry crops; prevent pre-harvest fall of the fruit; prevent the germination of tubers and root crops during their long-term storage; acceleration of rooting of cuttings during vegetative propagation; induction of flowering agricultural and ornamental crops, and in a number of other cases.

As regulators of plant growth in agricultural practice, various chemicals are widely used that belong to different classes of organic compounds. This, for example, is the sodium salt of benzoyl formic acid, used to increase the field germination of cotton seeds, increase its yield by treating the seeds before sowing with a 0.001% solution; α-naphthylacetic acid (alpha-NAA), used to spray tobacco seedlings before sampling with a 0.001-0.0015% solution to stimulate root formation; a combined preparation, which is a mixture of ethephon and chlorocholinchloride, used to combat lodging of winter wheat and rye by spraying plants in the tubing phase; Ketostim, a mixture of sodium salts of benzene (mono) diketocarboxylic acids, used for use on cotton to increase yield by moistening the seeds with a 0.005% solution; Nicofan - a mixture of growth substances and vitamins: metabolic products of the symbiotic sea buckthorn fungus, including vitamins A ₁ , A ₂ , B ₁ , B ₂ , biotin, nicotinic and pantothenic acids, giberellins A ₁ -A ₃ , methylene blue and toluene; increases productivity and resistance to diseases (Handbook edited by V.I. Martynenko and other Pesticides, M .: Agropromizdat, 1992, s.264-279). As follows from the above, plant growth regulators (herbicides, retardants, etc.) differ in toxicity and selectivity of action on individual plant species. Their great variety (plant growth regulators) is determined by their selective effect on certain types of plant crops (tobacco, cotton, wheat, rye, rice, flax, vegetables, fruit plants, ornamental plants), as well as their effect on productivity, improving their technological qualities increasing immunity to diseases.

In particular, the drug “Furolan, g” [2- (2-furyl) -1,3-dioxolane] is known as a plant growth regulator used for fruit crops (peach, plum, sweet cherry), cereals (rice, wheat), vegetable crops (sugar beet), contributing to an increase in their productivity, increase in their size, increase resistance to lodging, immunity to diseases (Handbook of pesticides and agrochemicals approved for use in the Russian Federation ”, M .: Publishing house“ Agrorus ”, 1999, p.178 -179).

A sharp reduction in the current volume of supplies to the country's agriculture of intensification means - machinery, fertilizers, pesticides, as well as a natural deterioration of the climate requires a significant increase in the biologization of agricultural production, among which the use of technologies of phytohormonal regulation of production processes in agriculture should take one of the most important places .

The theory of hormonal regulation of plants has almost a century of history - from the first morphophysiological experiments of Kholodny-Vent to modern ideas about the hormonal status of plants and its genetic determination. Recent discoveries in this area are associated with the discovery of natural phytohormones in plants - auxins, gibberellins, cytokinins, abscisic acid. Nevertheless, the scale of production and use of phytoregulators in the world and in our country has far from reached the expected values.

An object of the present invention is to provide a new plant growth regulator having an immunomodulatory, anti-stress effect.

The stated technical problem is achieved in that the new plant growth regulator (stimulator) is a complex based on derivatives of hydroxycinnamic acid, including chicoric acid, chlorogenic acid and caffeic acid in a mass ratio of 79-81: 14-16: 4-6, respectively, obtained by sequentially treating the air-dried plant material obtained from the aerial part of plants Echinacea purpurea, including extraction of the material with 90-96% ethanol with a mass ratio of ethanol and raw materials of 59-61: 1, respectively the next acidification of the extract with salicylic acid to a pH of not more than 5.0, separation of the plant inhibitors from the extract by further purification with a polymer sorbent and concentration of the extract by evaporation.

The technical task is also achieved by the growth regulating preparation, including the above growth regulating complex based on derivatives of hydroxycinnamic acid in a certain ratio, obtained by the above method, which is one of the claimed group of inventions with its content in the preparation of 0.009 - 0.11 mg / ml, and surface-active a substance based on ethoxylated alkyl phenols in a ratio of 1: 1.

The stated technical problem is also achieved by the fact that the above-described growth-regulating preparation is used for processing agricultural, ornamental, woody plants and shrubs.

Moreover, to increase the germination of seeds and increase the energy of their germination, the drug is added in an amount of 0.2 - 1.0 mg / ton. To increase the resistance of plants to adverse environmental conditions, improve root formation and increase crop yields, this drug (taken in the calculation of 0.1 - 10 mg / ha) is sprayed with plants in the period from full germination to the beginning of budding.

Stated as one of the inventions, a growth regulating complex based on derivatives of hydroxycinnamic acid (chicory, chlorogenic, coffee) performs numerous functions in the life of a plant cell, the importance of which is comparable in some ways with the role of phytohormones. In the literature there is evidence of the participation of chlorogenic and caffeic acids in the processes of rhizogenesis (Collection edited by M.N. Talieva, L.V. Runkova “Natural phytohormones and inhibitors”, 1978, p.25-32). However, the results of these authors are preliminary in connection with their pronounced dependence on the type of culture and on the physiological background of the studied plants. In addition, the effectiveness and direction of action of the drug depends on the concentration of specific acids used in the complex, as well as on its concentration in the drug.

And this feature of the action of these representatives of hydroxycinnamic acids requires further studies to elucidate the nature and mechanism of their action, the selection of optimal conditions, including the range of sensitive crops that show appropriate responsiveness to the used HCC (hydroxycinnamic acids). Numerous results have been obtained on models of animal systems, indicating the presence of the considered representatives of HCC - chicorye, chlorogenic and caffeic acids - immunomodulating, antibacterial, antifungal and antitumor activity (to varying degrees for each type of acid). In addition, chicoryic acid, the main component of the active substance of the claimed drug, has a powerful antiviral activity, realized through the inhibition of virus integrase (M.N. Zaprometov “Phenolic compounds, distribution, metabolism and functions in plants”, M .: Nauka, 1993, p.66-67; V.I. Kefeli “Natural growth inhibitors and phytohormones”, M .: Nauka, 1974, p.24, 93-101).

However, there is currently no evidence of a similar effect of chicory acid on plant viruses. There is also no direct evidence of the presence of a direct protective function in relation to plant pathogens in the examined GCC. Nevertheless, the information available in the above monographs regarding the relationship between derivatives of hydroxycinnamic acid and the activity of IAA oxidase, an enzyme that plays a key role in the growth processes of plant cells, as well as data on significant changes in the composition and content of phenolcarboxylic compounds (which include HCC) during the transition of the cell to the generative state and a possible connection with the transition to flowering allow us to consider these representatives of the class of derivatives of hydroxycinnamic acids from the point of view of the prospects of their use as means that can modulate in the right direction the physiological basis of growth and generative processes in plants. At the same time, the current situation of crop production in the face of increasing environmental pollution pressures, soil depletion and weakening of the protective properties of plants puts on the agenda the search for new ways to strengthen the adaptive mechanisms of the plant cell, increase their tolerance to adverse environmental conditions, and increase productivity , and in the field of ornamental crop production - in combination with the improvement of the decorative properties of crops.

Description of the invention

The active ingredient, or the active substance (AI) of the preparation according to the invention, is represented by a mixture of hydroxycinnamic acids (chicory acid: chlorogenic acid: caffeic acid in a mass ratio of them, for example, 80: 15: 5), isolated from the plant material - aboveground part Echinacea purpurea. The following is a description of a preferred embodiment of the invention from the claimed group relating to a method for producing a new drug.

I. Preparation

The procedure for obtaining the drug includes the steps of: isolating the active substance (AI) and creating the formulation. In turn, the stage of allocation of D.V. consists of two substages: primary processing of raw materials and fractionation of the extract. The specified production process includes the extraction of natural components from a plant source and, accordingly, is not associated with the implementation of the chemical conversion process.

The scheme for obtaining the drug (complex)

I. Obtaining the active ingredient (dv drug)

I.1. Preparation of plant material for the isolation of a complex of hydroxycinnamic acids and primary processing of raw materials

I.2. Extraction of fixed feed by polar solvent

I.3. Fractionation to Separate from Plant Inhibitors

I.4. Evaporation of partially purified enriched in AI extract

II. Obtaining a technical product in the form of a complex

I. A detailed description of the stages of the preferred method of obtaining AI growth regulating complex

I.1. Preparation of plant material for the isolation of a complex of hydroxycinnamic acids and primary processing of raw materials

Dry-air raw materials are obtained from the All-Russian Institute of Medicinal and Aromatic Plants (VILAR). The aerial part of Echinacea purpurea plants is collected manually in the second year of plant development. The raw materials supplied for use are accompanied by a quality certificate in which the supplier (VILAR) indicates the content of the amount of hydroxycinnamic acids in the dried raw materials (at least 2.5%). It should be noted that even higher requirements are imposed on Echinacea raw materials intended for medical use in the form of pharmaceutical preparations, which a number of suppliers (USA, Australia, etc.) certify not only by the total content of hydroxycinnamic acid derivatives, but also directly by level the main component is chicory acid (from 0.81%).

I.2. Extraction of fixed feed by polar solvent

Dry-air material is extracted with 96% ethanol (in the ratio of plant material to ethanol, for example, 1:60). The indicated ratio of extractable material to ethanol differs from traditional practice, which focuses on the use of 50 parts of ethanol per 1 part of plant material. This modification is the result of experimental selection of optimal methods for the extraction of components of AI (estimated by the amount of chlorogenic acid in the extract).

Next, the extract is decanted and 50 mM salicylic acid is added to acidify the extract (max. To pH 4-5) in order to enhance the stability of the active complex (primarily chicorye as well as chlorogenic acid) and increase the completeness of extraction of all three components of a.v.

The described extraction mode (including the extracting agent, the pH value and the nature of the acidifying agent) was selected based on the study and testing of different methods for isolating the HCC complex of interest and is optimal for the implementation of the purpose of the present invention, representing one of its aspects.

Thus, in the first stage of obtaining AI per 1 kg of raw materials, 60 l of ethyl alcohol and salicylic acid are used in an amount of 414 g (based on 6.9 g / l) per taken amount of raw materials.

When carrying out the extraction in accordance with the described procedure, it is possible to sufficiently completely extract a complex of phenolic compounds from plant materials, however, many by-products are extracted, including impurities of compounds of an inhibitory nature. This, of course, is its negative side and puts on the agenda the task of partial purification of the obtained extract. In this context, it should be noted that the contradictory nature of the literature data on the effect of HCC on rhizogenesis and plant growth may be explained by the fact that these authors, when isolating a complex or specific HCC representatives, did not get rid of phenolic compounds of inhibitory nature, but depending from the conditions of isolation, the level of such compounds and their ratio with stimulating compounds may vary, which, accordingly, affects the activity of the obtained extract. In order to reduce the negative impact of such compounds, the authors introduced a special stage of purification of the extract with a polymer sorbent, which allows separation of the activator complex of the cell and phenolic inhibitors, which previously was a serious problem due to the similar behavior of these compounds within the framework of various known in this field procedures for the allocation and purification of HCC.

I.3. Fractionation to Separate from Inhibitors

To reduce the level of impurities in the obtained ethanol extract, primarily the inhibitory effect, the authors developed a method for partial purification of an acidified extract to pH 4.5 by fractionation on a polymer sorbent. As the polymer sorbent, various sorbents are used, for example, crosslinked (co) polymers, such as a styrene-divinylbenzene copolymer (Polysorb), polyvinylpyrrolidone and others. The method of the present invention allows purification in the format of a batch procedure in which impurities, including compounds of an inhibitory nature, remain in the liquid phase, and the HCC complex of interest is deposited on a polymer sorbent (according to spectrophotometric evaluation of the active complex tester - chlorogenic acid deposited on the polymer sorbent) .

Elution is carried out with 10 l of ethyl alcohol, per 1 kg of raw material taken.

Variant of using 96% ethanol at the indicated stage of preparation of the drug is fundamental, because, as follows from a number of reports in the literature, chicory acid is sensitive to water content and undergoes rapid degradation at high humidity. A number of authors recommend the use of high concentration ethanol to increase stability, in general, of various HCCs, and, in particular, chlorogenic, which is characterized by instability during storage. So, for example, the chlorogenic acid preparation supplied by Fluka Chemica-Biochimica (cat. No. 25700) is regulated by the water content of not more than 3%, and the chicoryic acid-rich preparation of echinacea by Hammer Pharma is also limited by the water content of not more than 5% .

In addition, it is essential that chicory acid is poorly soluble in water at room temperature (only 0.5% solubility). And this fact additionally indicates the desirability of using high concentration of ethanol for desorption of the HCC complex.

I.4. Evaporation of partially purified enriched in AI extract

The extract after the desorption step in the partial fractionation procedure is evaporated, for example, in vacuum on a Buche R-500 rotary film evaporator to 1/10 volume.

II. Obtaining a technical product (Growth regulatory drug)

The technical product is obtained by combining ethanol solution (96% ethanol) in a concentration of 0.01-0.1 mg / ml with a carrier of the formulation — surfactant — monoalkylphenyl ethers of polyethylene glycol, for example, in a 1: 1 ratio.

The use of the preparation according to the invention for the treatment of plants

The preparation according to the invention can be used in amounts of from 0.1 to 10 mg / ha, the exact dose is determined by the specific direction of application and specific cultures, with the aim of:

strengthening adaptive properties, increasing crop yields: treatment is carried out by repeated spraying (2 times) of vegetating plants in the full germination phase - the beginning of budding with a working aqueous solution (1 ml of the drug in 10 l of water) in an amount of 0.1 to 10 mg / ha;

stimulation of root formation processes in plants by soaking the cuttings in a solution of the preparation according to the invention (1 ml of the product in 1 l of water) for a period of 10-24 hours;

flowering induction: a) corms are soaked in a working solution of the drug (1 ml of the product in 1 l of water); b) vegetative plants are sprayed with a working solution of the drug (0.1 - 10 mg / ha); in this case, the preparation according to the invention accelerates the beginning of flowering of decorative crops and increases the yield of elite products;

increase germination and increase the energy of seed germination by soaking the seeds at room temperature for 8-16 hours in a working solution of the drug (0.2 mg / t).

The following examples illustrate the technical result when using a growth-regulating preparation based on a complex of derivatives of hydroxycinnamic acid obtained in a specific way that make up the group of inventions.

Example 1

For example, the effect of the drug on the susceptibility of pine seedlings to the common fungus Heterobasidium annosum (pathogen of coniferous root rot) was investigated. Pine seeds previously soaked (20 hours) in water (control) and a solution of the preparation according to the invention at a concentration of 0.1 mg ai in 25 l, infected with the filtrate of the fungus. After 10 days of germination, seeds not treated with the drug were infected with a fast-growing strain of the fungus by 94%, slow-growing - by 72%. The treatment of seeds with the preparation according to the invention increased their resistance: the number of infected pine seedlings decreased: with a rapidly growing strain decreased by 2 times, and by 9 times with a slowly growing strain.

Example 2

The influence of seed treatment of cucumbers, tomatoes, sweet pepper with the preparation according to the invention on the growth and formative processes is studied. The treatment of cucumber seeds (1.25 mg / kg / 1.5 L of water) had an impact on the germination rate and germination of seeds, the length of the roots and sprout, as well as their weight. The increase in the yield of cucumbers amounted to 24.9%, the increase in the yield of tomatoes - 34%. The drug stimulates the growth of pepper plants, the process of leaf formation and dry matter content, contributes to the formation of larger fruits by size and weight (fruit length - 7.6 - 9.4 cm (control - 6.5 cm), fruit diameter - 4.4 - 5.3 cm (control - 3.9 cm), fetal mass - 152.6 g (control - 130 g).

Thus, the growth-regulating complex according to the claimed invention and the growth-regulating preparation based on it activate the processes of chlorophyll synthesis, growth and plant rhizogenesis, compensate for the deficiency of natural growth regulators, increase the adaptive capacity of the body in adverse conditions, act as a flowering inducer of plants, and exhibit indirect antifungal and antibacterial activity .

Example 3

The preparation according to the invention showed good results when used both for treating pea seeds (0.4 mg / t) and for spraying crops (0.1 mg / ha) during budding - the beginning of flowering.

The results show that he has a growth-stimulating effect, which is manifested by an increase in length (up to 34.7%) and mass of seedlings (up to 31.5%), plant height at the initial stages of development (up to 16-31%). A decrease in the rate of development of root rot in pea crops to 20% was recorded. Pea productivity increased by 7.9-11.0%. The treatment of seeds of spring wheat and spring barley (1 mg / t) and subsequent spraying in the tillering phase (2 mg / ha) stimulated the passage of the main phases of crop development: entry into the tube, heading, milk and wax ripeness, which came 10 days earlier, than in control. Two-time use of the drug (seed treatment and spraying of vegetative crops) increased the collection of wheat from 24.9 to 30.0 kg / ha, barley from 23.6 to 27.0 kg / ha. The drug had a positive effect on the quality of grain: it reduced humidity during the harvesting period, increased the protein content, and in the grain of wheat and gluten. The defeat of crops by root rot when using the drug decreased from 2 to 1 point.

Example 4

On ornamental plants of various taxa when processing the preparation according to the invention, the processes of rhizogenesis in cuttings, flowering and fruiting were studied. Soaking of hard-rooted cuttings (0.2 mg / l, control - water, 14 hours) of Japanese sakura, three-lobed almond, floribunda roses and hybrid tea, clematis stimulated root formation from 30 to 50%. Double treatment of the marigold with vegetative plants (0.1 mg / l) by the preparation of budding accelerated the development of plants. Full (100%) flowering in these plots occurred by June 30, while in the control variant there were only 27% of flowering plants by this time. In a Starina rose, after treatment with the preparation, flowering occurred 10 days earlier than in the control. The number of buds and flowers exceeded the control by more than 3 times.

Example 5

The preparation according to the invention accelerated the growth of roots and aerial parts of seedlings of Siberian larch and Scots pine when sprayed at a dose of 0.001 mg / L. Processing of seedlings of Siberian cedar led to an increase in both the height of the seedlings and their accumulation of biomass. Experienced seedlings had indicators of height that were 17-26% higher than the control, and the mass of their aboveground parts (the trunk with needles) exceeded the control by 19-34%. Spraying cherry trees (0.01 mg / l) with the preparation during the budding period increased the yield from 7% to 28% for Lebedyanskaya and from 8% to 16% for Kharitonovskaya. The cherry fruits in the experimental versions fully ripened, the fall of the ovary was minimal.

Example 6

Spraying blackcurrant bushes (4 mg / ha) during the budding phase with the preparation according to the invention increased the yield by 0.15-0.40 kg from the bush and reduced the susceptibility of powdery mildew by 33-37%, septoria by 19-42%.

Along with the growth-regulating properties, the preparation according to the invention exhibits immunomodulating and anti-stress effects. The use of the drug dramatically reduces the degree of damage to diseases such as late blight, alternariosis, rhizoctoniosis, peronosporosis, common scab, bacteriosis, fusariosis, brown rust, gray rot of strawberry, white rot of carrots, powdery mildew of roses and black currant. For example, treating winter wheat with a preparation reduces the rate of mildew sowing by 0% and brown rust from 52.3% to 4%; when growing cucumbers in both protected and open ground, the drug reduces the incidence of peronosporosis by 20-60%. It was found that the drug has an adaptogenic effect on plants in drought conditions. So, the biomass of Alpine penny, treated with the drug, was 53 mg versus 35 mg per plant in the control variant. In wheat plants treated with the preparation, photosynthetic potential increased during drought, and the work of a unit of assimilation surface was stimulated. The value of NPF increased from 0.30 to 0.41 mg / cm ² per day in the irrigated version and from 0.37 to 0.52 mg / cm ² per day under drought conditions, which determined the activation of biomass accumulation during treatment with the drug and increased stability plants to drought.

Claims (6)

1. A method of obtaining a plant growth and development regulator in the form of a complex of derivatives of hydroxycinnamic acid by sequentially treating air-dried plant materials from the aerial parts of plants Echinacea purpurea, including extracting it with a 90-96% standard with a ratio of ethanol and raw materials 59-61: 1 accordingly, subsequent acidification of the extract with salicylic acid to a pH of not more than 5.0, separation of the fraction of plant inhibitors from the extract by further purification with a polymer sorbent and concentration of the extract by evaporation . 2. The plant growth and development regulator, which is a complex of derivatives of hydroxycinnamic acid, including chicoric acid, chlorogenic acid and caffeic acid in a mass ratio of 79-81: 14-16: 4-6, respectively, obtained by the method according to claim 1. 3. The growth-regulating preparation, including the growth-regulating complex according to claim 2, with its concentration in the preparation of 0.009-0.11 mg / ml and a surfactant based on ethoxylated alkyl phenol in a ratio of 1: 1. 4. The use of a mixture of a complex of derivatives of hydroxycinnamic acid, including chicoric acid, chlorogenic acid and caffeic acid in a weight ratio of 70-81: 14-16: 4-6, respectively, obtained by the method according to claim 1, with a concentration in the mixture of 0.009-0, 11 mg / ml and a surfactant based on ethoxylated alkyl phenol with a 1: 1 ratio of the components of the mixture as a growth-regulating drug in the processing of agricultural ornamental, woody plants and shrubs. 5. The use of a mixture of a complex of derivatives of hydroxycinnamic acid, including chicoric acid, chlorogenic acid and caffeic acid in a weight ratio of 70-81: 14-16: 4-6, respectively, obtained by the method according to claim 1, with a concentration in the mixture of 0.009-0, 11 mg / ml and a surfactant based on ethoxylated alkyl phenol with a 1: 1 ratio of the components of the mixture as a growth-regulating drug introduced into the seed soaking medium in an amount of 0.2-1.0 mg / t. 6. The use according to claim 4, characterized in that the mixture is used for spraying vegetative plants in an amount of 0.1-10 mg / ha.