RU1811365C - Agent for plant growth regulation - Google Patents

Abstract

An agent comprising a mixture of 2-piperidone and 1 - {(2- (4-hydroxyphenyl) ethanoyl -2-piperidonyl or 1- {3- (4-hydroxyphenyl) propanoyl -2-piperidone in a ratio of 1 : 1-1.5, respectively. The product may also contain 2-7 x 10 cells of microorganisms of the species Rhizoblum or Streptomyces per 1 mg of active substance. Processing the plants or soil with this product can increase productivity and reduce the incidence of plants. The product not only stimulates plant growth , but also inhibits the growth of harmful microorganisms and promotes the development of beneficial ones. -22 tab.

Description

The invention relates to means for regulating plant growth and may find application in agriculture.

The purpose of the invention is to increase yield and reduce the incidence of plants.

Important conditions for achieving the goal are accelerated root (plant) growth, rapid propagation of leguminous bacteria of the genus Rhizoblum and actinomycetes of the genus Streptomyces which are useful for growing crops (soil), and suppression of the predominance of bacteria of the genus Pseudomonas and pathogenic fungi that cause painful damage.

Careful studies of physiologically active substances for use in agriculture, taking all these points into account, have led us to discover that the problems mentioned above can be solved by using a physiologically active substance, including the N-acyl lactam compound and 2-piperidone. which highly stimulates plant root growth and regulates painful lesions. It was also found that the use of this substance together with leguminous bacteria or actinomycetes of the genus Streptomyces has a good effect on the population of these beneficial microorganisms in the soil.

... The invention relates to physiologically active substances for use in agriculture, comprising, as an active ingredient, a mixture of the M-acyllactam compound represented in the general formula

nol (sn2) asoy),

about

y-7 where n 1.2

and 2-piperidone.

The invention also relates to a physiologically active substance for use in agriculture, which

With

WITH

00

00

about eating

includes N-acylactam compound, 2-piperidone and legume bacteria Rhlzoblum jap. Further, the present invention relates to a physiologically active substance for agricultural use, which comprises an N-acylactam compound, 2-piperidone and actinomycetes of the genus Streptomyces.

N-acyllactam and 2-piperidone can be mixed together in powder form, or a uniform mixture can be obtained by dissolving the two compounds in a suitable solvent.

There is no particular restriction on the choice of solvent type, and any solvent capable of dissolving the two compounds can be used for this purpose. Unusually, methanol, ethanol, acetone and ethyl acetate are used. When catalytic reduction is selected for the production of N-acyllactam, 2-piperidone may be added to the reactor in advance.

The physiologically active substances of the present invention are preferably used in the form of a solution. For uniformity, but can also be used as a mixture with a suitable carrier such as talc, cyclodextrin, dextrin, vermiculite, diatomaceous earth and silicon powder.:

The appropriate amount used may vary depending on the type of plant, soil environment, and the purpose being pursued. Typically, an aqueous solution with a concentration of 0.1-10 mg / L is used in an amount of about T L / m for use in soil, and an aqueous solution with a concentration of 0.01-0.1 mg / L is used in an amount of 1-10 t / 10 acre for use on leaves.

Substances of the invention are preferably used in the seed stage (e.g., in a potted state before planting or within two weeks after planting), but can also be used at any other convenient time.

Alternatively, the compounds of the invention can be applied to the soil before planting, or they can be added, hydroponically, to a tank of water, to the nutrients or fertilizers used.

The physiologically active substance, including the compound of N-acyllactam, is examined in detail below. 2-piperidone and leguminous bacteria of the genus Rhizobium.

As an example, legume bacteria. Rhizobium genus. used in the present invention can be cited among others

Rhizobium japonlcum, R. leguminosarum and R. trifolll.

The use of such leguminous bacteria in combination with the compound of N-acyllactam and 2-piperidone is especially effective in the cultivation of legumes.

For the purposes of the present invention, any legume bacteria cultured in the usual manner can be used. They are

can be used in the form of a culture solution, granules obtained after centrifugal separation, or in the form of a mixture with a suitable carrier, as described above (e.g. talc).

The physiologically active substance of the present invention containing legume bacteria is particularly effective when applied around the roots of each plant. . .

Suitable quantity will vary by type.

plants, environment, soil and other factors.

The following is a physiologically active substance, including the compound Nacillactam, 2-piperidone and actinomycetes

genus Streptomyces.

The physiologically active substance of the present invention using actinomycetes of the genus Streptomyces in combination with a compound of N-acylactam and 2-piperidone is particularly effective in accelerating the actinomycete population in the soil.

6 as an example of genus actinomycetes used in the present invention

Streptomyces can be cited, among others, Streptomyces olivocheomogenes, S. phacochromogenes, and S. grfscolus.

Any actinomycetes of the genus may be used for the purposes of the present invention.

Streptomyces cultured by conventional methods.

The method of application and the appropriate amount are the same as in the case of the substance described above containing

bean bacteria. Both soil and leaf applications can be used and there are no restrictions on the time of application.

The physiologically active substances of the present invention have the following effects:

- accelerate the growth of plant roots;

- legume bacteria of the genus Rhizobium and actinomycetes of the genus Streptomyces proliferate in the soil, which are useful microorganisms for plant growth;

- suppress the predominance of bacteria of the genus Pseudomonas and pathogenic fungi in the soil, which are microorganisms that cause painful damage to plants.

Thus, the inventive tool provides normal crop growth, yields high yields, especially legumes. In addition, a substance containing legume bacteria; or actinomycetes of the genus Streptomyces alleviate soil salinity with the aforementioned beneficial microorganisms, which cannot be achieved by conventional methods. The result is a significant increase in crop yield (with legumes) and the prevention and reduction of plant diseases.

All microorganisms used in the present invention are well known and can be obtained from the following storages:

ATCC: Collisions of American Cultures, Rockville, USA

IFO: Fermentation Institute, Osaka, Japan

NCTC: National Crop Collection, Central Health Laboratory, London, England

BUCSAV: Institute of Biology. Czechoslovak Academy of Sciences, Prague, Czechoslovakia

NCIB: National Collection of Industrial Bacteria. Tori Research Station, Aberden, Scotland

CBS: Centralburean voor Schimmelcultures, Baaen, The Netherlands.

PIA: All-Union Research Institute of Antibiotics, Moscow, USSR. The method of obtaining compounds 1 and 2

1. (Compound 1)

10 ml of a 5% by weight to volume methanol solution of sulfuric acid was added to 1 g of 4-hydroxyphenylacetic acid, after which the mixture was kept at 80 ° C for 6 hours under reflux conditions. After completion of the reaction, ether was added, the ether layer was separated and washed with 1N. a solution of sodium hydroxide and water in this order were dehydrated and then the ether was distilled off, whereby a methyl ester was obtained.

 The methyl ester was dissolved in dimethylformamide in a 10-fold amount: an equal molar amount of anhydrous potassium carbonate was added to this solution, followed by an equal molar amount of benzyl chloride, and this solution was kept at 80 ° C for 6 hours under conditions heating under reflux. After completion of the reaction, ether was added, the ether layer was washed with 1N. hydrochloric acid solution and water in

the indicated order, and then the ether was distilled off, whereby a benzyl ether was obtained.

1 n a solution of potassium hydroxide in a 10-fold amount was added to benzyl ether, this mixture was heated to 180-190 ° C and cooled when the oily substance disappeared and the mixture turned into a white suspension.

The white suspension was filtered, dissolved in excess ether, washed with 1N. a solution of hydrochloric acid and water in this order and dehydrated, and then the ether was distilled off, in

5 resulting in a white powder.

A 1.3-fold molar amount of thionyl chloride was added to the white powder, and the resulting mixture was kept at 80 ° C in

0 for 3 hours under reflux. After the completion of the reaction, thionyl chloride and gaseous hydrochloric acid were removed under reduced pressure, and a pyridine solution of 2 times the molar amount of 2-piperidone was added. A chlorine-calcium tube was installed and the mixture was kept at 80 ° C for 5 hours. After completion of the reaction, ether was added, the ether layer was separated, washed with 1N. hydrochloric acid solution, 1 N. a solution of sodium hydroxide and water in this order and was dehydrated, and then the ether was distilled off, resulting in

5, 1.02 g of (4-benzyloxyphenyl) ethanol -2-piperidone was obtained.

Then this substance was dissolved in ethyl acetate and this solution was kept at room temperature for 5 hours,

0 by stirring in a hydrogen atmosphere and using 5% palladium carbon as a catalyst. After completion of the reaction, ethyl acetate was distilled off, whereby 0.70 g was obtained.

5 1- 2- (4-hydroxyphenyl) ethanoyl -2-piperidone.

2. (Compound No. 2)

10 ml 5% in terms of weight to volume

A 0 methanol solution of sulfuric acid was added to 1 g of 3- (4-hydroxyphenyl) propionic acid, and this mixture was kept at 80 ° C for 6 hours under reflux conditions.

5 After completion of the reaction, the ether layer was separated and washed with 1N. sodium hydroxide solution and water in that order. dehydrated and then the ether was distilled off, whereby a methyl ester was obtained.

The methyl ester was dissolved in dimethyl formamide in a ten-fold amount, then an equal molar amount of anhydrous potassium carbonate and an equal molar amount of benzyl chloride were added to this solution. This mixture was kept at 80 ° C for 6 hours under reflux conditions. After completion of the reaction, ether was added to this mixture, the ether layer was washed with 1N. hydrochloric acid solution, 1 N. a solution of sodium hydroxide and water in this order, after which the ether was distilled off, whereby a benzyl ether was obtained.

1 n a 10-fold amount of potassium hydroxide solution was added to benzyl ether, after which the mixture was heated to 180-190 ° C and cooled when the oil substance disappeared and the mixture turned into a white suspension.

The white suspension was filtered and dissolved in excess ether, the resulting product was washed with 1N. ether solution, the resulting product was washed with 1 N. a solution of hydrochloric acid and water in that order, then dehydrated and the ether distilled off, resulting in a white powder.

A 1; 3-fold molar amount of thionyl chloride was added to the white powder, and this mixture was kept at 80 ° C for 3 hours under reflux conditions.

After the completion of the reaction, thionyl chloride and gaseous hydrochloric acid were removed and, under reduced pressure, a pyridine solution of 2: a multiple molar amount of 2-piperidone was added, a calcium chloride tube was installed and this: the product was kept at 80 ° C for 5 hours

After completion of the reaction, ether was added, the ether layer was separated, washed with 1N. hydrochloric acid solution, 1 N. sodium hydroxide solution and water in the indicated order, 1N. a solution of sodium hydroxide and water in this order and was dehydrated, after which the ether was distilled off, whereby 1.42 g of 1- 3- / 4-benzyloxyphenyl / propanoyl-2-piperidone was obtained.

Then this substance was dissolved in ethanol and the resulting solution was kept at room temperature for 6 or 5 hours, stirring in a hydrogen atmosphere and using 5% palladium-carbon as a catalyst. After completion of the reaction, ethanol was distilled off,

and the solution was extracted with a mixture of water and methanol (1: 1), which allowed to obtain 0.74 g of 1-3 / 4-hydroxyphenyl / propanoyl -2-piperidone.

Compounds No. 1 and No. 2 thus obtained were identified by H-NMR spectroscopy.

Thus, each microorganism referred to in the following examples will be given its location number.

Example. Physiologically active substances of the present invention (molar ratio 1: 1) were dissolved in ethyl acetate, obtaining experimental solutions

certain concentrations, as indicated in the table. 2.

Each test solution (2 ml) was added to a piece of filter paper (70 mm in diameter) placed in a Petri dish,

the solvent was distilled off under reduced pressure, 2 ml of sterile water was added, and Brasslca rapal seeds were planted. (25 pcs.) And kept at 25 ° C in the dark.

For control, the same operation was repeated, but using only ethyl acetate. After 48 hours, the length of the grown roots was measured and the degree of root growth was calculated from the difference in the data of the control samples. The results are given in table. 1.

Example 2. Twenty-five pieces of germinated rice seeds were placed on a 2% agar in a Petri dish, with each coleoptil pointing up and kept at 25 ° C for two days, spraying

water.

An experimental solution of 50% acetone (1 ml) was added using the microsyringe between the first leaves of the sprouts with the concentration indicated in the table. 2. and continued

cultivation.

For control, the above procedure was repeated, but using only 50% acetone.

The results are presented in

tab. 2 ..

Plant height is expressed with the size of the control group after 24 hours, taken for 100.

PRI me R 3. Each physiologically

the active substance of the invention (1: 1 molar ratio) was dissolved in ethyl acetate in a conical flask and the solvent was distilled off under reduced pressure.

Separately, each of the species indicated in the table. 3, up to 50 ml of a liquid medium containing 2% malt extract was inoculated. Static cultivation was continued at 25 ° C for 4 days and the resulting mycelium was collected, which was dispersed in

sterile water, which gave an experimental microbial solution.

The culture medium of the composition indicated in the table. 4 (50 ml each) was dispensed into test flasks prepared previously (100 ml capacity, each containing 5 mg of test sample), each flask was sonicated in the tank to completely disperse the test samples. 1 ml of the biological test solution obtained above was inoculated and the static culture was continued at 25 ° C. for 10 days. Then, the weight of dry biological cells was measured and the degree of change in their weight was calculated. For control, the same procedure was repeated, but using ethyl acetate alone.

The results are given in table. 5.

From the results given in table. 6. It is clear that the physiologically active substances of the present invention explicitly regulate the propagation of fungi.

Example 4, Physiologically active substances of the invention (molar ratio 0.2) were dissolved in acetone to a concentration of 10 mg / L, a disc for antibiotic testing (8 mm in diameter) was placed in each solution and the solvent was evaporated, thereby obtaining experimental discs .

An agar medium containing 2% malt extract was placed in a Petri dish, water was removed from the surface, the experimental disk obtained earlier was placed in the medium, and 0.05 ml of the biological solution obtained from species No. 4 in the table was inoculated. 3 in the same manner as in example 3.

Static cultivation was laid at. 25 ° C for 10 days, on the fourth day and subsequent days (on which growth was observed), the area of biological growth on the surface was measured and the degree of growth of the area was calculated.

The same control procedure was carried out, but acetone alone was used. The results are shown in table. 6.

From the table. 6 it is clear that the physiologically active substances of the present invention are not fungicidal substances, the growth of fungi in the initial stage reaches a maximum on the 5th day, and then drops sharply.

PRI me R 5. Each of the species listed in the table. 7, were cultured at 30 ° C. using the growth medium shown in Table 8. Immediately after confirmation of the grown cells, the cells were kept on the surface of the beveled growth medium at 4 ° C.

as the main culture for subsequent experience.

In a 100 ml conical flask was placed 0.5 ml of an ethyl acetate solution (1000 mg / L) of the physiologically active substances of the present invention (molar ratio: 0.5). the solvent was distilled off and 50 ml of broth liquid medium was added.

One loop of the main culture obtained above was inoculable in 10 ml of liquid broth medium and uniformly dispersed. 0.5 ml of this dispersion was inoculated into a conical flask.

The same control procedure was repeated, but ethyl acetate alone was used. Cultivation was continued at 30 ° C for 48 h and the number of grown cells was counted using a Petrof-Hausser counting chamber. The results are given in table. 10.

As can be seen from the table. 9, the agents of the present invention are effective in controlling the growth of bacteria.

EXAMPLE 6 Each active substance of the present invention (molar ratios of 0.1, 1 and 10) was added to a stead of the composition below to a concentration of 0.1 mg / L. One loop of microorganisms shown in table. 10 was suspended in 10 ml of sterile water, 0.1 ml of the resulting suspension was inoculated to the above agar medium and cultivation was continued on Petri dishes at 25 ° C.

On the seventh day, the total number of grown cells was counted and their shape examined. The results are given in table. eleven.

Example. Wagner's vegetation vessels (1/5000) were filled with a mixture of soil and compost (1: 1) and fertilizers were applied so that the ratio -P20s teO was 0.5-0.5-0.5 / vessel.

Five Gliclne max. Were planted in each prepared vessel. with three leaves (total 15 pcs.).

After 10 days, the seedlings predicted and left at each place three seedlings of the same height. 14 days after planting, 20 ml of the alcohol solution of the prototype indicated in Table 1 was evenly distributed around the roots. 12, and the plants were left to grow, watering them with water and following insects and diseases.

74 days after planting, the number and weight of nodules and roots were measured; the measurement results are given in table. thirteen.

The biomass concentration in the above cell suspension was 1.4 x 107 cells / ml. In the case of tests Nsisfc 6-9

one ml of a suspension of cells was added to 20 ml of an active agent solution (the number of cells was 7 x 10 cells) mg of active agent). In case of Test No. 10, 10 ml of a cell suspension was added to 20 ml of an active agent solution (the number of cells was 7 x 108 cells) mg)

As can be seen from the table. 13, the mass of root nodules and the number of leguminous bacteria increased with the use of the invention.

An example. This example illustrates that the use of the physiologically active substances of the present invention greatly facilitates the propagation of Streptomyces genus actinomycetes.

One loop of the species listed below was inoculated to the beveled nutrient medium of a composition containing 1 g of 4 g of yeast extract, 10 g of malt extract, 4 g of glucose, 20 g of agar, which was then incubated at 30 ° C.

One loop of the thus grown cells was icoculated to a liquid medium of the same composition and cultivation was continued at 30 ° C for 72 hours to obtain a main culture.

Separately, ethanol solutions of the physiologically active substances of the present invention (molar ratio: -0.5) were diluted with sterile water to obtain solutions with a concentration of 1.0 and 10 mg / L, and each of the resulting solutions (2 ml) was placed in a Petri dish .

18 ml of agar medium was added to the Petri dish, to which 0.1 ml of the main culture was inoculated, after which cultivation in the cup was continued at 30 ° C.

For control, the above procedure was repeated, but the same amount of ethanol was added. On the seventh day, the number of colonies formed was counted and the results are shown in table. 14.

(I) Streptomyces canus IFO-12752 (ATCC-12237 and 19737;

CBS-475.68; PIA-1017)

(Ii) Streptomyces Fradlae IFO-12773 (ATCC-10745 and 19760;

CBS-498.69; PIA-1040)

PRI mep 9. The ethanol solution of each physiologically active substance of the present invention (molar ratio 1.5: 1) was diluted with water and a test solution with a concentration of 10 mg / L was obtained.

Separately, the wild type of a species of the genus Strepromyces, collected in the field, was cultivated for 4 days using the aqueous composition indicated in Table 1. 10. The cell concentration in the culture fluid is 2 x 109 cells / ml.

To 1 l of this culture solution was added 100 ml of the test solution obtained above.

One part of this active substance containing microbiological cells was mixed with ten parts of diatomaceous earth to obtain a microbiological sample for a field experiment.

This field sample was applied to 50 plants of cucumber (Cucumis satlvus), which were grown in the field for 50 days.

For control, only diatomaceous earth was added to the damaged area of the other 50 plants. We monitored the progress of painful injuries before harvesting and, at the last stage, counted the number of plants that produced the crop. The results are given in table. 15. An example. The test solution obtained in Example 9 was diluted with water to a concentration of 0.2 g / l and applied to 100 tomato plants growing in the field 10 days after planting (application in soil in an amount of 1 l / m2).

The state of painful lesions was examined on the 60th day after administration, which are given in table. 16. The number of diseases was determined by the presence or absence of damaged areas of the terrestrial part, the degree of stunting of leaves and stems, and the state of fruiting.

Example I. Pot experiments were conducted to determine the effect of the physiologically active substance of the present invention on soybean yield.

The conditions for soybean cultivation and use of the substance are given below.

(Soybean cultivation)

Main fertilizer: N-PaOS-feO 1.5-1.5-1.5 (g / pot) Planting time: July 1986

Harvest time: but in 1986 (Use of the substance)

(1) Type of substance: physiologically active substance No. 1 (molar ratio: 1)

(2) Concentration and amount: 1 mg / L,

1 l / m2, (3) Application time: 15 days after

landing

(4) Conditions for planting in a pot: three rows,

1/2000 acre pot, four plants per

pot. The results are given in table. 17.

Example 12. Definition of action

physiologically active substance

the present invention on potatoes. Below

potato cultivation conditions are given

and the use of the substance.

fifteen

. 1811365

16 table

The effect of the agent of the invention on root growth

Note: The degree of root growth was determined by the following equation.

The degree of root growth (%)

 (Medium, measurements of the experimental zotsa) - (Medium, measurements of the control zone) in (Medium, measurements of the control zone)

Substance No. 1: Compound No. 1 + 2-piperidone

Substance No. 2: Compound No. 2 + 2-piperidone

Table 2

The effect of the agent according to the invention on rice seedlings

17

Table The composition of the medium for growing microorganisms

Tablab Effect of the agent of the invention on the development of microorganisms

The degree of change in the mass of biological cells was calculated by the following equation:

The degree of change in the mass of biological cells (%) Dry mass in the experimental zone (mg) - dry mass in the control zone ---------------------------- ----------- x Yuo Suha mass in the control zone (mg)

1811365

18 table 3

Tablab Effect of the means of the invention on the development of microorganisms

The degree of biological growth of the area was calculated by the following equation:

The degree of biological growth of the area

Area growth in the experimental zone (cm)

Area growth in the control zone

Table

The microorganism cultures used in the experiment

Tablab Composition of a nutrient medium for microorganisms

Potato Extract Pressed Yeast Liver Extract

M sa extract

Thioglycolic Acid Medium Glucose Glycerol

Agar

Distilled water pH

The potatoes were peeled (100 g), cut into cubes about 1 cm, boiled in 500 ml of tap water for 30 minutes, cooled and separated from solid particles.

The liver (50 g) was finely chopped, cooked for 30 min in 150 ml of tap water, cooled and separated from solid particles.

200 g

30 g

25 g

5g

South

5g

15 g

15 g

l getting 1 liter) 7.0

Table E The effect of the agents of the invention on the development of microorganisms, table. 7

Soil extract; the soil (1 kg) was extracted with 500 ml of water at 121 ° C for 30 minutes, the mixture was left overnight and the filtrate was diluted to 1000 ml

The developmental impact of the agents of the invention

Table 10

Table 11

Note. The total number of cells is expressed as a factor to the value of the untreated samples.

The shape of the cells is shown in the bottom column, R; stick shape B; bacteroid.

The value in parentheses indicates the molar ratio of the agents of the present invention, accelerates the propagation of legume bacteria with a factor of about 1.5-3, and also enlarges the cells into a bacteroid.

Table 12 The composition of the active substances used in the experiment

Experience number

bean bacteria

1

2

3

4

6

7

8

9

10

Legume bacteria II; the cells obtained in example 6 by culturing species (II) at 25 ° C. for 10 days were suspended in sterile water to give an absorption of 0.3 at 660 mm.

Continuation of the table. eleven

Prepared sample solutions

e bacte and

additives (10 g / l)

2-piperidone

Compound No. 1

Substance Ms 1 of this invention

(molar ratio 1: 1}

Substance No. 2 of this invention

(molar ratio 1: 1)

2-piperidone

Compound No. 1

Substance No. 1 of this invention

(molar ratio 1: 1)

Substance No. 2 of this invention

(molar ratio 1: 1)

The effect of the agents of the invention on the development of Streptomyces

Table 13

Table 14

27

The effect of the agent of the invention on cucumber plants

Test sample

Diatom Earth

No

Compound. Number 1

Compound No. 2

Agent Mg 1

Agent number 2

Agent number 1 streptomycetes

actinomycetes Agent N ° 2 + streptomycetes actinomycetes

The effect of the agent of the invention on a tomato plant

Prototype

Substance of the invention:

substance No. 2 Comparative examples:

control.

2-piperidone

compound number 2

The effect of the agent of the invention on soybean yield

1811365

28 table 15

Plants capable of producing yield,%

4

sixteen

28

thirty

43

47

76

82

Table 16

Incidence of plants. %

5.1

18.15

19.6

15.3

Table 17

Table 18 The effect of the funds according to the invention on potato plants

Table 19 The effect of the funds according to the invention on the defeat of potatoes

Table 20

31

Agent molar ratio

This agent is not applicable.

1811365

32 table 21

Table 22