SU1707015A1 - Carbamoyl-derivatives of alkanolamines as regulators of plant growth - Google Patents

Abstract

The invention relates to carbamoyl derivatives, in particular alkanolamines of the total f-ly RO-C (0) -NH- (CH2) n-0-C- {0) NRiRa, where R is C-C-alkyl, phenyl; RI H or CH3: R2 chlorhexyl, naphthyl, chlorophenylsulfonyl, phenyl, mono- or dichlorophenyl, mono- or dimethyl-, trifluoromethyl-, nitro, methoxyphenyl; n 2 or 3, as plant growth regulators that can be used in agriculture. The purpose of the invention is the creation of new, more active substances of the specified class from available raw materials. The synthesis is carried out by the reaction of the compound f-ly RO-C (0) -NH- (CH2) n-OH, with isocyanate in an aprotic solvent medium or with phosgene, followed by treatment with an appropriate amine. The new compounds are more effective than cartoline in increasing the frost resistance of winter wheat, as well as the drought tolerance of wheat barley and seed by increasing osmotic cell pressure and stimulating synthetic processes in plants under conditions of moisture deficiency. 11 tab. n 2 or 3, which can be used as plant growth regulators. The closest to the proposed compounds in structure are their aromatic analogs - derivatives of O. N - diaoylcarbamate of the general formula (Qr-N-COO-0-HCOOC R and c from N. with 3 iso

Description

where R is OH or H,

possessing the properties of growth regulators

plants (cortolin and kinetin).

However, known compounds, including kartolin, have a number of drawbacks that limit their use: their activity as plant growth regulators is not high enough; relatively scarce and expensive starting materials, such as m-nitrotoluene and m-aminophenol, are used to produce these compounds. ; (during the preparation, intermediates that are not stable during storage — arylhydroxylamines, 0- (m-alkoxycarbonylaminophenyl) chlorocarbates, are formed;

The purpose of the invention is the search for new plant growth regulators that would be obtained from less scarce and expensive raw materials and exceeded in activity (as regulators) known plant growth regulators.

Compounds of formula (I) about the quality of plant growth regulators are proposed ..

The compounds of formula (I) are synthesized from available raw materials — alkanolamines, alcohol, amines, and phosgene — in two ways, starting from the compounds obtained by known methods

ROCONH (CH2) nOH. (II) exposing the compound of formula (II) to an isocyanate in an aprotic solvent medium or by stimulating the compound of formula (II) with phosgene and then treating the resulting chlorogol ester with an appropriate amine.

The compounds obtained by these methods are listed in Table. 1. They are new and not described in the literature. Compounds of formula (I) with R - H are stable crystalline substances, and with R - alkali - viscous oils. All compounds of formula (I) are poorly soluble in water and in common solvents; they can be used with the addition of surfactants in the form of aqueous dispersions or in the form of wettable powders to powder the seeds before sowing.

As shown by the results of the primary toxicological studies conducted on the example of compound (I) (code name Kartolin-2), this group of substances has low toxicity for warm-blooded living animals (acute toxicity LDso for white rats, -1000 mg / kg).

Example 1, To a solution of 0.005 mol N-alkoxy (eryloxy) carbonylalkanolamine of formula (P) in 15 ml of dry ether at 20 ° C, let the flow of phosgene slowly. By

at the end of the reaction (TLC control, 3 h), excess phosgene and hydrogen chloride are removed by stripping with nitrogen. 10 ml of ether are poured and 0.01 mol of the corresponding amine in 10 ml of ether is gradually added with vigorous stirring. The mixture is kept up, filtered, the solvent is evaporated, the residue is washed with hexane and the compound of formula (I) is obtained. Compounds 1,2 and 13 are obtained by this method (Table 1).

Example 2. To a solution of 0.04 mol of the compound of formula (II) in 60 ml of dry ether is added 0.1 ml of triethylamine and

With vigorous stirring, a solution of 0.04 mol of isocyanate in 40 ml of ether is added. The mixture is stirred for 2 hours at room temperature. The precipitated product is filtered and washed with a mixture of ether - hexane. Compounds of formula (I) (R1 - H) are obtained. Compounds 3-12 and 14-21 were obtained by this method.

The yield of the compounds of formula (I) obtained in examples 1 and 2 is 605 70%,

The structure of the compounds of formula (I) is proved on the basis of yes and other elemental analysis, IR and PMR spectroscopy,

In the IR spectra of these compounds, the absorption bands of the groups are observed, CM V KW): С 0 at 1730-1660: NHCO at 1550-1500: СОО at 1240-1190: (in CCU): NH at 3480-3420.

PRI me R 3. In the qualifiers, tests

5, a cell culture of parenchymal tissue of tobacco grown under strictly conditioned conditions on modified Schonk and Hilde-Brandt medium is used. The tests are carried out at

0 concentrations of substances. The activity of substances is evaluated according to the degree of their influence on the growth of the cell mass and the amount of carbon emitted by the cells in the process of respiration. Selection criteria are obtained when

5 study the nature of the effect of reference substances on cells. The duration of the experiment is 120 hours. As the main reference, take kartolin in the concentration of M, additional - kiietin in the concentration of M,

0 The test results are shown in Table. 2 as a percentage of the control (an object without drugs),

A series of compounds of formula (I) show activity characteristic of cytokinins.

5 purine plots taken in high concentrations, when the inhibition of carbon dioxide excretion by cells is inhibited significantly more actively than their growth. Therefore, to assess the degree of activity of the proposed compounds, it is convenient to use the difference in the criteria characterizing cell growth and respiration.

The greater the difference, the higher the activity. All proposed compounds, except for compounds 4, 8, 10, 18, and 19, have a significant amount of this indicator, and compounds 1, 3, 5, 7, 13, 16, and 21 have activity comparable to or exceeding the activity of the standard, caroline. Compounds 4, 8. 10, 18, and 19 have a significant stimulating effect on both recorded parameters, which is characteristic of cytokinins at lower doses.

Since all the studied compounds exhibit to some extent cytokinin activity, in the future they will be in special tests that assess the degree of this activity, as well as their protective properties.

EXAMPLE 4 The cytokinin activity of compound 1 was evaluated by determining its effect on the germination of salty seeds, which were in a state of thermo-induced rest (keeping for 1 day at 30 ° C). The results are shown in Table. 3

As a view but from the table. 3, compound 1 in this test shows an effect comparable to kinetin at a concentration three orders of magnitude lower, while certholine is inactive in this case.

EXAMPLE 5 The cytokinin activity of compound 1 was evaluated on experiments to study its effect on the intensity of the trans- formation of ten leaves cut from three-week-old wheat plants. The assessment is carried out by taking into account the amount of water evaporated over 24 hours of leaves immersed in the solution of the test preparation. In this test, compound 1, like kartolin, does not possess cytokinin activity.

This experience shows that, in contrast to cytokinins, the studied compounds do not intensify transpiration in the leaves, which should contribute to the manifestation of a protective effect on plants under drought conditions. The results are shown in Table. four.

PRI me R 6. Further testing for cytokinin activity was carried out with seven of the cucumber. Cucumber seedlings are grown in a thermostat at 27 ° C for 7 days. The seeds are separated and put into Petri dishes on filters moistened with a solution of the test compound in water. In the control filters are moistened with water, in the reference experiment - with a solution of kinetin. The seeds were incubated for 24 hours at 27 ° C in the dark, then exposed to light for 3 hours, after which chlorophyll was extracted with acetone and its content was determined on a spectrophotometer at a wavelength of 665 im.

In tab. Figure 5 shows the data on stimulating the formation of chlorophyll by the drug - 5% as a percentage of the control (water containing no drugs).

Compounds 1 and 16 are not inferior in strength to kinetin, which is ten times higher in concentration.

0 EXAMPLE 7 Compound 1 shows a high cytokinin activity in various biochemical reactions characteristic of the cytokinin action substances described in examples 7 and 8.

5 When studying the effect of compound 1 on nitrate reductase activity, work was performed on isolated embryos of puppets (Agrostemma githago, Z).

Seeds are soaked for 12 hours in cups.

0 Petri on distilled water at 30 ° C in the dark. Then, embryos are isolated from seeds and incubated in water, in solutions of different concentrations of the natural cytokinin analog — 6-benzylaminopurine and in connection with 5 denineni 1. The embryos are incubated in the dark at 30 ° C for 18 hours. WITH.

Nitrate reductase activity is determined in the in vitro system by the method of Ewens and

0 Nason with some modifications. Compound 1 stimulated nitrate reductase 250% (for kartolin 110%).

The cytokinin activity of compound 1 was established by induction of the synthesis of nitrate5 reductase in isolated embryos of dolls. This is the participation of compound 1 in the regulation of cytokinin-type activity of the genetic apparatus of plant cells. Kartolin in this experiment does not show a cytokinin effect. Example The protective effect of compound 1 on the protein synthesis apparatus is studied on barley plants. For experiments, leaves are taken from plants that have been subjected to drought without treatment and with preliminary treatment with synthetic cytokinins under the conditions of Example 12. As a control, leaves are taken from plants grown under conditions of

0 normal soil moisture. The benchmark is kartolin.

Barley leaves are ground in liquid nitrogen and homogenized in a buffer with high ionic strength (200 mmol tris-acetate, pH 8.5; 60 mmol KCI; 30 mmol MgCla: 250

5 mg / oh of syrup; 4 mmol / -mercaptoethanol).

The homogenate is centrifuged sequentially at 15,000 g for 10 minutes and at 18,000 g for 20 minutes. Post-mitochondrial

g of the supernatant add triton X-100 to a final concentration of 1%, layered on a pillow of 1.75 M sucrose in buffer. containing, mmol: Gris-acetate 40. pH 8.5: KCI 20; MgCl2 S; / -mercaptoethanol 4. and centrifuged at 42 thousand rpm D for 90 min. The sediment of ribosomes is suspended in a buffer containing, mmol: Tris-acetate 10, pH 7.9; KCI25; MgCl25, / J-mercaptoethanol 4, and fractionated in a sucrose concentration gradient (10-35%) in a SW-40 rotor of a centrifuge for 70 minutes. Thereafter, the tubes are punctured, the fractions are collected in distilled water, and the optical density is measured at 260 nm.

Results:

Option Percent Policy

Control45

Drought18

Drought connection 169

Drought + Certerolin63

Treatment of barley with compound 1 leads to an increase in the proportion of polysomes in ribosome preparations. isolated from the leaves of fused plants, Dol by polis under the action of drought decreases in leaves from 45 to 18% and rises under the action of compound 2 to 69%,

The obtained data testify to the .j, ui-oM deigdBpi of compound 1 to the SS apparatus. High to-.1 polisom indicates the functional activity of the apparatus of protein synthesis fj d. St x. what determines the intensity of protein synthesis about them and reflects the overall functional activity of the leaves, which plays a sooty role in the productivity of plants.

EXAMPLE 9 In laboratory conditions, the effect of the compounds of formula (1) on the factor of plant resistance to drought is determined by the ability of the seeds of wheat seeds of variety Bezos I to grow on solutions with increased osmotic pressure. This test is the ability of compounds of formula (I) to increase osmotic pressure (OD) in cells and stimulate synthetic processes in plants under conditions of moisture deficiency, which determines the resistance of plants to drought as well as it causes the development of a more powerful primary root system.

The neck poses soy osmotic pressure of 16 atm is created by dissolving sucrose. The results of the action of the compounds of formula (I) introduced into the solution of azarose are given in table. b in percentage of the control - the amount

seeds germinated on sucrose solution with the same osmotic pressure but without the addition of compounds of formula (I).

These attempts connect 1 and 16 in strength

The protective action is superior to caroline. The results are shown in Table. 6

Example 10: The effect of the proposed compounds on the resistance of wheat leaves to moisture deficiency and lowered temperatures is evaluated. The degree of damage to the leaves is assessed by the rate of electrolyte loss due to disrupted cell membranes. The lower the electrolyte yield, the lower the degree of disturbance

5 i.e. the effectiveness of the drug is higher, the seeds of wheat cultivar Illichivka are grown for 8 days on the tested preparations, then the cut leaves are kept for 24 hours over native calcium chloride or for 1 day at a temperature of

-4 ° C, Ds-ice are immersed in distilled water and, by its final electrical conductivity, the amount of electrolytes released.

5 The control, in percentage, to which the data obtained are attributed, are dead leaves.

The results are shown in Table. 7. Concentration of substances 10 mg / l.

0 As can be seen from the above results, all of the investigated compounds are comparable or superior to kartolin in terms of their protective effect.

PRI me R 11. Assess the ability

5 of compound 1 to protect wheat germ from low temperatures. For this, the seeds of wheat cultivar Illichivka are soaked for 4 hours and germinated for 5 days in a solution containing the preparation (control 0 is water). Then the seedlings quenched 5 days at 9 ° G. and pomooazhivat 5 days at a temperature

-10 ° C. An indicator of sustainability is the proportion of seedlings capable of producing new roots and a green leaf after freezing. as a percentage of the control. The results are shown in Table. 8. As can be seen from the table. 8, compound 1 at a dose of 0.1 mg / l has a protective effect exceeding the effect of kartolin. VZRTOGO

0 at a dose of 1.0 mg / l.

Example 12. Next, the effect of compounds on drought tolerance on the conditions of the phytotron with the maximum approximation to the real situation is estimated, in a field. The experiments were carried out on the barley of the Favorite variety, which is relatively little responsive to the effects of kartolin. Barley plants are grown in a phyto greenhouse complex. Drought conditions are created in the fourth and sixth stages of organogenesis within 2 weeks.

Before transferring to the drought regime, the first part of the plants is treated with compound 1 at a dose of 1 kg / ha, and the second - with kartolin at the same dose and the third - with water. The control plants are grown at a soil moisture of 70% of full capacity.

The test results are shown in table.

The treatment of plants with compound 1 contributes to the preservation of the harvest of barley at a higher level in comparison with the drought variant without treatment and with the treatment with kartolin.

The data obtained indicate a more effective protective action during the drought period of compound 1 in comparison with the preparation kartolin.

PRI me R 13. The effect of the proposed substances on frost resistance of winter wheat varieties Mironovsk 808 is studied. Frost resistance of wheat is assessed by freezing plants in refrigerators on a phytotron. Before sowing into the soil, seeds are coated with a wettable powder containing a wetting agent, an adhesive. filler and 20% active ingredient. Before freezing the plants undergo two hardening phases: Phase I - 10 days at 2 ° C, Phase II - 3 days at -5 ° C. Then step by step them

five

live in cabinets at temperatures of -13, -16. -18 and -21 ° C. After thawing, the plants are planted in the soil and after 5 weeks of frost resistance fall, which is expressed in percent of the surviving plants or the number of initial plants taken for freezing.

The results are shown in Table. ten.

For comparison, table. 11 shows similar data for the drug kartolin.

From tab. 10 and 11, it can be seen that compound 1 is more effective in increasing the frost resistance of winter wheat than kartolin.

Claims (1)

Invention Formula Carbamoyl derivatives of alkanolamines of general formula vosgasuposo / R 4R Ii. where R is C1 C4 alkyl. phenyl; R1 is hydrogen, methyl; R is chlorhexyl, naphthyl, chlorophenylsulfonyl, phenyl, phenyl, containing as substituents 1 or 2 chlorine atoms, methyl, trifluoromethyl, nitro or methoxygroup; n 2 or 3, as plant growth regulators. table 2 Table 3 Table 4 Table 5 Table 6 - chlorocholin chloride, Table 7 Table 8 Table 9 Table 10 Table 11