RU2337543C2 - Agent for seed greensprouting stimulation and increasing sprout resistance to moisture deficiency, and method for its obtaining - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: method for obtaining an agent stimulating seed greensprouting and increasing sprout resistance to moisture deficiency is proposed. First hydroquinone is oxidised with hydrogen peroxide in the presence of ferric iron salts. The resulting acid is then neutralised with alkali. After that the mixture is incubated at a temperature of 50-65°C during 5-6 hours, and subsequently the unreacted hydroquinone content is measured. If its content exceeds 20%, a fresh portion of hydrogen peroxide is added and a reaction is maintained until the hydroquinone content does not exceed 10-15% of the total weight of the agent, or the reaction mixture is blown with air during 2-3 hours. An agent stimulating seed greensprouting and increasing sprout resistance to moisture deficiency is described. The main functionally active component of the agent is an oligomer containing several phenolic ring and functional OH groups, having a molecular weight of approximately 3 kDa.

EFFECT: increased quantity of sprouts; weight and height of plants.

2 cl, 1 tbl, 6 dwg, 5 ex

Description

The invention relates to agriculture, namely to crop production, and can be used to increase seed germination, seedling growth rate and increase their resistance to lack of moisture.

In the germination of seeds of grain crops, the limiting factors are humidity, its maintenance at a constant level, as well as sterility from pathogenic fungi and microorganisms. In the seed kernel there is all the necessary supply of nutrients, and until they are completely consumed, the seedlings can do without top dressing.

Various substances are used to stimulate seed germination.

A known method of producing liquid suspended peat humic fertilizers (RU No.2001038, class C05F 11/02, 1993.10.15), which can also be used to accelerate seed germination.

However, the resulting preparations have an unstable composition, so it is difficult to standardize, i.e. select a dose for optimal stimulation of seed germination. And in a non-optimal concentration, it can produce the opposite effect, i.e. inhibit the process of seed germination.

Good results for stimulating seed germination are given by soaking before germination in solutions of gibberrelin or calcium pangamate with a concentration of 50-100 mg / L (RU No. 2112344, CL A01C 1/00, 1993.10.15).

Known drug Katapol used for presowing treatment of seeds. It is a complex of a copolymer of N-vinylpyrrolidone and crotonic acid with alkylmethylbenzylammonium (RU No. 2133568, class A01N 25/10, 1999.07.27).

However, in the latter case, the substance is not a characteristic natural component for plants and, despite its non-toxicity in mice, the question of possible long-term effects on the human body remains open.

The closest to the claimed drug analogue that stimulates the germination of seeds is potassium humate grade "A" produced by ZAO "TPK Technoexport", Sergiev Posad, Moscow region. under the trade name "Energen". (TU 2387-020-423 15284-00, No. of state reg. 27-7410-0143-1-mark-A). This drug, as written on the package, contains salts of humic acids, salts of silicic acids, macro- and microelements.

However, due to the wide variety of chemical composition of humic acids obtained from various natural sources, such preparations are difficult to standardize. In addition, the seed germ already contains trace elements and other nutrients necessary for growing without additional addition of trace elements to the medium at the stage of seedling growth up to 12 days of age. Thus, the presence of a multicomponent nutrient composition in humates is not strictly necessary to stimulate seed germination.

It is known that the complex effect of huminate on the properties of sandy podzolic soil was expressed in increasing the total biomass of plants and soil moisture capacity (ttp: //www.humina.ur.ru/doc/flexcom.doc) and improving its structure (http: // www. moarcc.aris.ru).

A known method of producing polyoxyphenylene ethers (RU No. 2224737, class C07C 43/295, 2004.02.27), which consists in the fact that the hydroquinone is oxidized with hydrogen peroxide at a molar ratio of these reagents 1: (1.5-2), respectively, in the presence of ferrous salts, and then the neutralization of the formed acid with alkali.

However, in the preparation obtained according to the aforementioned patent, the content of free unreacted hydroquinone is not regulated, it can be up to 60% of the total preparation. This causes a toxic effect on seed germination.

In the proposed general structure of the base compound, there are two phenyl rings joined by oxygen.

where R = —OH, —S — SO ₂ —ONa; x = 1, 2; M = —OH, —O — C ₆ H ₄ —OH.

In addition to it, the final preparation contains iron and sodium cations, the remaining free hydroquinone and, possibly, a quinone-hydroquinone compound.

The obtained compounds possessing a combined antioxidant and antihypoxant effect were used only in medicine.

The objective of the invention is to provide a drug to stimulate seed germination and increase the resistance of seedlings to moisture deficiency.

The technical result that can be obtained using the proposed drug is an increase in the number of seedlings, mass and height of plants.

In the period of seed germination, the stimulating role of humus and humic acids is determined by their positive effect on maintaining the moisture content of the grain and stable supply of water to it.

The substances of humic acids, which determine the moisture capacity of the substrate of germination of seeds and the seeds themselves, are substances of aromatic nature that give these acids and humus a dark color.

These substances can be replaced by synthetic preparations, in particular, substances obtained by the oxidation of hydroquinone with the formation of polyoxyphenyl ethers.

The drug, obtained as described in patent RU No. 2224737, as well as humic acids, is highly soluble in water and poorly in alcohol, forms a precipitate with trivalent aluminum and other heavy metals. Common with humic acids is also good solubility in alkalis and sorption on the surface of silica gel, sand, cotton, at neutral and acidic pH.

Since this compound is formed by oxidation of hydroquinone in the presence of ferrous iron and hydrogen peroxide, the final product has a dark color and a absorption spectrum similar to humic acids in the visible region. Some other physico-chemical properties of the preparation prepared according to patent RU No. 2224737 are also similar to humic acids, in particular, in the absorption spectrum in the visible region. (J. Cieslewicz. Comparison of the chemical composition of sediments in lakes in catchments forested with beech and pine / in collection Earth and Environmental Sciennces // Acta Universitatis Latviensis, v.692, 2005, pp. 7-18).

The proposed drug to stimulate seed germination and increase the resistance of seedlings to moisture deficiency is obtained as follows.

Hydroquinone is oxidized with hydrogen peroxide in the presence of ferrous salts, and then the formed acid is neutralized with alkali, the mixture is incubated at a temperature of 50-65 ° C for 5-6 hours, then the content of unreacted hydroquinone is measured if its content is more than 20%, add a fresh portion of hydrogen peroxide and carry out the reaction to a hydroquinone content of not more than 10-15% of the total weight of the whole preparation.

To reduce the content of unreacted hydroquinone, the reaction mixture is purged with air for 2-3 hours. During the entire course of the reaction, the content of free hydroquinone is monitored by one of the known methods, for example spectrophotometric.

Then the mixture is dried in a freeze dryer to a concentration corresponding to a 1.5% -3.5% solution, or until dry.

The liquid form is well stored and more convenient for dosage when using. A small precipitate formed during storage can be completely dissolved in water.

The resulting preparation for stimulating seed germination and increasing the resistance of seedlings to moisture deficiency is a substance containing several components. The main functionally active component is an oligomer having a molecular weight of 3-4 kDa, including several phenolic rings and functional OH groups.

Confirmation of this is the data of thin layer chromatography (Figure 1), spectrometric characteristics in the visible region (Figure 2) and data of electrophoretic separation of the drug by molecular weight in polyacrylamide gel (figure 3).

Figure 1 presents the result of thin-layer chromatography of hydroquinone, the drug "Energen", the drug according to patent RU No. 2224737 and the proposed drug, obtained using air purging of the reaction mixture. Chromatography was performed on 20 × 10 cm cellulose plates (Watman) in a solvent system of methanol: dimethylformamide: water 75:15:10. Components showed iodine vapor. Hydroquinone has an R _{f of} 0.95. In "Energene" and the proposed drug there is no hydroquinone, while in the preparation prepared according to patent RU No. 2224737, it is present in an amount greater than 20%. In the preparation "Energen" and in the proposed preparation, the bulk of the substance remains at the start. The second component of the proposed drug has a mobility lower than that of hydroquinone, R _f 0.86 (indicated by arrow).

The spectrometric characteristics in the visible region in FIG. 2 indicate the similarity of the substances presented (there is no pronounced absorption maximum), all substances are equally colored — they have a dark brown color at a pH of about 7. This indicates that they have conjugated bonds between phenolic rings and oligomeric structure. The difference in the spectrum when KOH is added to the Energene preparation and the proposed preparation — the shift of the absorption spectrum to the long-wavelength region is less intense than in the case of the preparation according to RU patent No. 2224737. This is consistent with the data of thin layer chromatography (Figure 1) and confirms the different contents of hydroquinone in the preparations.

The data of polyacrylamide gel electrophoresis (Figure 3) of two preparations - proposed and patent RU No. 2224737 - confirm the similarity and difference in composition. The similarity is the presence of units at the start (indicated by arrow). The difference in composition: in the preparation according to patent RU No. 2224737, there are about three bands with different molecular weights from 3 to 5 kDa, in the proposed preparation one component with a molecular weight of about 3 kDa is found.

The possibility of implementing the invention is confirmed by examples, but is not limited to them.

Tests of the proposed drug was carried out in comparison with the drug "Energene", since this drug is widely used at present as a stimulator of seed germination.

Example 1. Dissolve 5 g of hydroquinone in 100 ml of distilled water until complete dissolution. With poor dissolution, ethanol is added in a ratio of 1: 2 v / v (aqueous solution - ethanol), the volume is adjusted to 100 ml. Then add 0.5 g of iron sulfate and 150 ml of 3% hydrogen peroxide, mix, adjust the pH to 6.5 with 0.1 m NaOH solution. The mixture is incubated with stirring at a temperature of 50 ° C for 5 hours.

The content of unreacted hydroquinone is measured. Its content is 27%. A fresh portion of 3% hydrogen peroxide in an amount of 70 ml is added. The content of hydroquinone after incubation under the same conditions is 12% of the total dry weight of the entire preparation. The reaction stops when the evolution of foam (oxygen) ceases.

Example 2. The method is carried out similarly to that described in example 1 except that when the content of unreacted hydroquinone is equal to 23%, air is passed through the reaction mixture to the complete absence of hydroquinone. The determination of hydroquinone content is carried out spectrophotometrically by shifting the absorption spectrum in the visible region with the addition of a 0.1 M KOH solution using a calibration graph (Figure 1).

Example 3. The treatment of seeds with the proposed drug at a concentration of 0.05%.

The indicated concentration approximately corresponds to that for the Energen preparation in terms of the content of colored aromatic substances. The drug "Energen" was used according to the attached preparation instructions: 5-10 drops are diluted in 50 ml of water.

Seeds are soaked in a ratio of _^1/2 volume of seeds / volume working solution products in plastic cups of 150 ml. In the control experiment, water is used instead of solutions. In this case, the seeds are completely immersed in liquids in three experiments.

The glasses are kept open at room temperature. After a day, seed swelling was observed in three experiments. After another day, the drying of the remaining free liquid in each cup is observed. In this case, the concentration of colored substances in the solution of the experiment with the drug "Energene" and the experience with the test drug increases.

On day 3, the onset of germination is observed in the experiment with the test drug and Energen, and in the control (soaking in water) this process has not yet begun.

In the first two experiments, there is no specific smell of decomposition and fermentation and germination is observed.

In the photograph (Fig. 4), it is seen that in the experiment with the proposed preparation, a noticeably greater root formation and greater germination of seeds are observed than in experiments with the Energena solution and with water.

After 10-12 days, the leaves of all three experiments are aligned in height. However, the number of leaves grown to the same height in the experiment with the proposed drug is 40-45% more than in the experiment with Energen and 55-65% more than in the experiment with water.

Example 4

In plastic cuvettes 350 × 220 × 60 mm seeds were soaked. The thickness of the dry seed layer is 20 mm. The concentration of the drug solution in water is 0.05%. A solution of the drug covers the seed layer.

Figure 5 shows that in comparison with the control (seeds soaked in water) in the experiment with the proposed drug, a significantly greater root formation in the context of the seed layer.

The table presents the results of determining the germination of wheat seeds, determined by a standard method (taking into account the germination of 100 seeds on filter paper).

Wheat germination Control (water) (in%) With the drug "Energen" (in%) With the proposed drug (in%) 68.0 72.5 94.5

Example 5

The effect of the proposed drug on the resistance of seedlings to a lack of moisture when germinating seedlings of wheat varieties Moskovskaya 358.

Before sowing seeds in sand cuvettes, the proposed preparation is added in a concentration of 0.07%; after alteration, wheat seeds were sown. The control did not contain the proposed drug.

Cultivation is carried out at room temperature and lighting with fluorescent lamps. 12-day-old seedlings do not water for 3 days. As a result, a significant number of plants wilt in the control. Figure 6 shows that the number of unfading wheat seedlings in the experiment is greater than in the control.

Thus, a preparation is proposed for stimulating seed germination. The presented examples confirm the claimed effectiveness and usefulness of the proposed drug.

The drug used is low toxic (data obtained in experiments on mice), and after its incubation with seeds it loses toxicity.

The proposed substance and the method for its preparation can be used in the industrial production of plant seedlings in order to obtain biologically active components from them, for example, wheat germ oil, vitamin E, phytosterols and lipase and proteinase inhibitors. In addition, the drug can be used to increase resistance to moisture deficiency.

Claims (2)

1. A method of obtaining a drug to stimulate seed germination and increase the resistance of seedlings to moisture deficiency, which consists in the oxidation of hydroquinone with hydrogen peroxide in the presence of ferrous salts, and then neutralization of the formed acid with alkali, the mixture is incubated at a temperature of 50-65 ° C for 5-6 hours, after which the content of unreacted hydroquinone is measured, if its content is more than 20%, a fresh portion of hydrogen peroxide is added and the reaction is carried out until the content of hydroxy it is not more than 10-15% of the total weight of the whole preparation is carried out or purging the reaction mixture with air for 2-3 hours. 2. A preparation for stimulating seed germination and increasing the resistance of seedlings to moisture deficiency, the main functionally active component of which is an oligomer comprising several phenolic rings and functional OH groups having a molecular weight of about 3 kDa obtained by the method according to claim 1.

Images