CS274442B2 - Agent for plants' anthers sterilization - Google Patents

Abstract

The agent for sterilization of plant anthers contains active substance in combination with diluter. Terpenoid compounds of the general formula I serve as active substance, where A stands for<IMAGE>or their mixtures. Plant treatment with this agent is carried out in the fifth and/or sixth stage of organogenesis. The agent is used in cultivation and seed growing industry.

Description

BACKGROUND OF THE INVENTION The present invention relates to biology and agriculture, and more particularly to a composition for sterilizing plant anthers which can be used in breeding and seed production.

Today, the challenge of intensifying agriculture, in particular increasing the profitability of cereals, feed, vegetables and technical cultures, is being addressed worldwide by the widespread use of first generation hybrids. Hybrids differing from parental forms can be obtained by crossing with higher productivity (25-30%) and better product quality.

There is a method of producing new hybrids that corresponds to a system of cytoplasmic pollen sterility - regenerated fertility. The basis of this method is continuous (12 to 14 years) and complicated selection work, which includes obtaining sterile analogs, sterility fixators and fertility restorers. The methods of sterilizing plant anthers with chemical sterilizing agents (gametocides) are particularly promising. The use of gametocides is considerably more economical than the use of a cytoplasmic pollen sterility system, since there is no need to develop such forms as sterile analogs, analogues for fixing sterility in female forms and for regenerating fertility in male forms. The seeds of the first generation hybrids can practically be obtained both during the breeding of the starting forms and also in the organization of their technical production.

There are currently found about 200 compounds having gametocidal activity and their chemical structure belongs to different classes of chemical compounds. Gametocides in treated plants must ensure complete sterility of the pollen while maintaining the ability of the egg cells to live and ensure a sufficiently high ability (at least 70% of control) to produce the testicle under free pollination. Their phototoxicity and warm-blooded toxicity values shall be minimal.

Methods of sterilizing cereal anthers are known (L. Dzh. Nikell, Regulators of Growth Growth, Primenenie in Rural Farming, Moscow, Ed. Kolos, 1984, pp. 28-31;

SU 906457), which consists in treating plants with sterilizing agents such as 2-chloroethylphosphonic acid (Ethrel), maleic hydrazide, di- (polyfluoroalkyl) phosphoric acids and their salts and others. Treatment of plants with sterilizing agents is carried out in V or VI. stage of organogenesis (according to F.M. Kupermann).

In the V stage of organogenesis, the processes of the formation and differentiation of flowers begin. Towards the end of this stage, neoplasms of sporogene archesporium tissue arise. During this stage germs of stamens, pistils and petals are formed. In Stage V differentiation of the rod embryo in the rod and pistil occurs. Stadium VI. is characterized by the process of the formation of the flower (micro and macro sporogenoza). Separate mononuclear pollen grains are formed at this stage (F.M. Kupermann, Morphofiziologia rastenii, Miskva, Higher School, 1973, pp. 30-36).

Further, a method of sterilizing grass anthers (GB.A. 1567153) is known, which comprises treating the grasses with a sterilizing agent for a period between the appearance of the second stem and the ear. As the sterilizing agent, heterocyclic compounds are used, the main representatives of which are 2-carboxy-3,4-methanpyrrolidine or 2-methoxycarbonyl-3,4-methanpyrrolidine. The compounds are used in combination with diluents and surfactants.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method of sterilizing anthers in a wide variety of cultures with a high degree of sterilization, while maintaining a high seed capacity for free pollination.

The object is solved by the fact that in the proposed method of sterilization of anthers of plants by treatment with a sterilizing agent in conjunction with a diluent in the fifth and / or sixth stage of organogenesis, according to the invention the terpenoid compounds of the general formula

CHj H

in which AND indicates -CH-CH, -CHCH, 1 , -CH, CH-CHCH,, -CH, CH, CHCH, 2 _{(/ X} / 2 2 2, 2 C (CH ₃ ) ₂ C (CH ₃ ) = CH ₂ C (CH3) ₂ -CH, CH, CHCH, 2 2 2 , -CH, CH, CHCH, -, -CH, CH, CCH, - ² 2, 2 ^{2 2} J ² C (CH ^{3) 2} C ₀ CH 3 C (CH 3) ₂ C (CH3) ₂ OH -CH = CHC = CH- HC _{(CH3) 2} or theirs mixtures.

The sterilizing agent may be used in conjunction with any known diluent known in the art. It is expedient to use it in conjunction with water as a 0.1 to 2% aqueous emulsion. The plants which are treated with said sterilizing agent are preferably grasses or sunflowers.

The composition according to the invention makes it possible to achieve the sterility of male plants (98 to 100 Ss) and to obtain a high seed formation capacity (over 70%). In order to achieve a high sterilization degree of anthers under unfavorable climatic conditions, treatment of the plants with a sterilizing agent in the fifth and / or sixth stage of organogenesis (according to Kupermann) is repeated.

The process is carried out as follows.

Plants such as winter wheat and spring wheat, diploid and tetraploid rye, triticale, millet, sunflower are treated with a sterilizing agent, the terpenoid compounds of the formula

CHj H

in which

And it means

-CH-CH, -CHCH, 12 ^ -2

C (CH3) 2

CH CH - CH CH ² LX ^ ² C (CH) ₂

-CH, CH, CHCH2,2,2

C (CH3) ₂ OH

-CH ₂ CH ₂ CHCH ₂ -CH ₂ CH ₂ CHCH ₂ C (CH ₃ ) -CH ₂

C (CH ₃ ) ₂ OCOCH 3

-CH ₂ -CH ₂ -CCH ₂ R (CH ₃ ) ₂

-CH = CHC = CH HC (CH 3) ₂ or mixtures thereof.

The terpenoid compounds may be used in combination with any suitable diluent. Suitably, water is used as the diluent. Preferably, a 0.1 to 2¾ aqueous emulsion of said compounds is used.

Suitable surfactants may be added to the ready-to-use solutions as desired. When applied to plants, any known adjuvants such as wetting agents, dispersing agents and enhancing agents are usually added to the ready-to-use solutions.

The sterilizing agent is applied to the plants by various treatments, such as liquid spraying and air spraying (aerosols). Treatment of plants with a sterilizing agent is performed at the fifth and / or sixth stage of organogenesis (according to Kupermann). The dosage of the sterilizing agent depends on the type of compound, the culture being treated, the stage of treatment and natural climatic factors. In order to ensure a high sterilization effect under adverse climatic conditions, repeated treatment of the plants in VI. stage of organogenesis. The total dose of the sterilizing agent is 0.6 to 20 kg / ha.

All of the terpenoid compounds of the invention for use as a sterilizing agent were tested for toxicity in animal experiments. The results of the tests showed that the compounds are slightly toxic or practically non-toxic.

For example, the LD-g is 4300 mg / kg for 6-terpenol and 3700 mg / kg for o-pinene.

All terpenoid compounds of the invention are found in nature and are derived from coniferous resin.

The gametocidal activity of the sterilizing agents of the invention was determined by field trials in different soil-climatic zones with partial areas of 10 m in size, with two, three to four replicates. Each sterilizer has been tested for at least 5 years.

The development of organogenesis stages is monitored cytologically. Treatment of plants with a sterilizing agent is carried out at the beginning of the fifth stage of Kupermann's organogenesis.

After the ears have grown, isolate the largest ears and the others in parchment bags. For wheat and triticale, individual isolates are used. At the rye, the ears of up to 7 different side-by-side plants are sealed in a single bag. For millet, each lath is isolated separately. The percent sterility (X) for wheat, rye, triticale and millet is determined by the formula number of seeds fertilized per isolate of treated plants

X = (1-) 100 number of seeds fertilized in one isolate of untreated control plants

The number of grains in the uninsulated spikes of the control plants is taken as 100% free pollination.

In order to obtain reliable data, 20 to 25 wheat and triticale isolates are used each time, 10 to 15 rye and millet isolates are used.

To treat chemical sterilization of sunflower pollen, 45 treated plants were used for each compound, 15 plants were isolated for self-pollination, 15 plants were pollinated with a mixture of pollen collected from 20-25 isolated dressings and 15 plants were left free to pollinate to control deployment of pollen-treated achenes.

The sterility of the pollen of sunflower plants is assessed by pollen fertility and germination, by the morphological peculiarities of the male germ cells and by the ability of the pathogens to be deployed in pollinating the treated isolated plants with pollen untreated plants. The life of CS 274442 B2 egg cell potency is determined after seed formation in treated plants with free pollination of untreated plants.

It is desirable to treat plants in clear, windless weather.

All compounds enter the plant tissue within 4 hours. In the event of rainfall during these 4 hours, the plants must be re-treated in VI. stage of organogenesis.

For a better understanding of the invention, the following working examples of the process according to the invention are given.

Examples 1 to 6

Mironovskaya 808 winter wheat plants are treated by spraying a 2¾ aqueous emulsion of the following sterilizing agents at stage V of organogenesis (according to Kupermann) using a portable sprayer: lp-menthen-8-ol, lp-menthenyl-8-acetate, 3-carene, l, 4 (8) -p-menthadiene, 1,8-p-menthadadiene or p-cymol.

0.1% wt. C 12 -C 14 calcium alkylbenzene sulfonate. As an adjuvant, 0.01 wt. dodecyl sulfate. The consumption of the product is 12 kg / ha.

As a control, plants treated with a diluent without a sterilizing agent.

The test results are shown in Table 1. Similar results are obtained when treating plants in VI. stage of organogenesis.

Table 1

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free 'pollination. Control 40.6 0.0 42.6 100.0 1 0.0 100.0 40.1 94.1 2 0.0 100.0 38.8 91.1 3 0.0 100.0 39.7 93.2 4 0.6 98.5 40.2 94.4 5 3.8 90.7 41.8 98.1 6 0.7 98.3 39.9 93.7

Examples 7 to 13

Plants of spring wheat of the species Moskovskaya 35 in VI. stage of organogenesis treats with 1% aqueous emulsion of the following sterilizing agents: 6-terpenol, 1p-menthenyl-8-acetate, 3-carene, 1,4 (8) -p-menthadiene, 1,8-p-menthadiene, p-cymol optionally 06-pinene.

The emulsion contains an emulsion of 0.1% by weight. % C 12 -C 14 calcium alkyl benzene sulfonate and adjuvant 0.01 wt. Ν, Ν-dimethylformamide. The consumption of the product is 8 kg / ha.

Control plants treated with a diluent without sterilizing agent serve as control plants.

The test results are shown in Table 2. Similar results are obtained for the treatment of plants in stage V of organogenesis.

Table 2

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 29.4 0.0 31.2 100.0 7 0.0 100.0 27.2 87.2 8 0.0 100.0 28.6 91.7 9 0.2 99.3 29.0 92.9 10 0.0 100.0 28.1 90.1 11 0.8 97.3 30.0 96.2 12 1,2 95.9 27.3 87.5 13 0.0 100.0 28.7 91.0

Examples 14 to 20

The spring wheat of Botanicheskaya 4 is treated with a 1% aqueous emulsion of the following sterilizing agents in stage V of organogenesis: 3-carene, β-cymol, β-pinene, οϋ -terpenol, 1,4 (8) -p-menthadiene, 1 8-p-menthadiene or 1p-menthenyl-8-acetate.

The emulsion contains 0.1 wt. % C 12 -C 14 calcium alkyl benzene sulfonate as emulsifier and 0.01 wt. tetrahydrofuran as an adjuvant. The consumption of the product is 6 kg / ha.

As a control, plants treated analogously to Example 1 are used.

The test results are shown in Table 3. Similar results are obtained when treating plants in VI. stage of organogenesis.

Table 3

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 28.9 0.0 30.9 100.0 14 0.0 100.0 27.8 90.0 15 Dec 1,2 95.9 30.1 97.4 16 0.0 100.0 29.2 94.5 17 0.0 100.0 28.4 91.9 18 0.0 100.0 29.5 95.5 19 Dec 0.8 97.2 27.3 88.3 ^{20 May} | 0.0 100.0 29.5 95.5

Examples 21 to 23

Plants of diploid rye of Chulpan species in VI. stage of organogenesis (according to Kupermann) using a portable sprayer, treats a 1¾ aqueous emulsion of 1,4 (8) -p-menthadiene 1,8-p-menthadiene or p-cymol.

The emulsion contains 0.1 wt. of C 12 -C 14 calcium alkyl benzene sulfonate and 0.01 wt. dodecylsulfate. The consumption of sterilizing agent is 10 kg / ha. As a control, plants treated analogously to Example 1 are used.

The results of the tests are shown in Table 4. Similar results are obtained for the treatment of plants in stage V of organogenesis.

Table 4

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free. pollination Control 44.1 0.0 49.6 100.0 21 0.8 97.8 47.8 96.4 22nd 0.3 99.2 43.2 87.1 23 0.0 100.0 47.4 95.6

Examples 24 to 27

Tetraploid rye plants of Ukraina tetra are treated as in Example 1. A 1¾ aqueous emulsion of the following sterilizing agents is used: 2-pinene, t-terpenol, terpenylacetate, 3-carene.

The consumption of sterilizing agents is 8 kg / ha. As a control, plants treated with a diluent without sterilizing agent. The test results are shown in Table 5.

Table 5

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 28.0 0.0 36.8 100.0 24 0.0 100.0 36.3 98.6 25 0.4 98.6 33.8 91.8 Control 36.1 0.1 40.2 100.0 26 0.0 100.0 32.8 81.6 27 Mar: 0.1 99.6 37.3 92.8

Examples 28 and 29

As in Example 1, diploid rye plants of Kharkovskaya 55 are treated with a 1% aqueous emulsion of the following sterilizing agents: terpenylacetate or 3-carene.

The consumption of sterilizing agent is 6 kg / ha. The test results are shown in Table 6.

Table 6

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 31.5 0.0 39.4 100.0 28 0.1 99.7 35.2 89.3 29 0.6 98.1 36.7 93.1

Examples 30 and 31

Plants of the triticale of the PRAG 109 type are treated, as in Example 1, in stage V of organogenesis, with a 2% aqueous emulsion of terpenylacetate or 3-carene sterilizing agents. The consumption of sterilizing agent is 12 kg / ha.

The test results are shown in Table 7. Similar results are obtained when treating plants in VI. stage of organogenesis.

Table 7

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 41.3 0.0 45.8 100.0 30 0.0 100.0 40.9 89.3 31 0.3 99.3 37.6 82.1

Examples 32 and 33

The triticale plants of Amfidiploid 206 were as in Example 1 of VI. stage of organogenesis is treated with a 2% aqueous emulsion of terpenylacetate or 3-kerene sterilizing agents. The consumption of the product is 12 kg / ha.

The results of the tests are in comparison with the control shown in Table 8. Similar results are obtained for the treatment of plants in stage V of organogenesis.

Table 8

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 23.4 0.0 25.2 100.0 32 0.4 97.8 23.0 91.3 33 0.0 100.0 23.8 94.4

Examples 34 to 39

Millet plants of Mironovskaya 94 are treated, as in Example 1, in V. stage of organogenesis with a 1% aqueous emulsion of the following sterilizing agents: terpenylacetate,

3-carene, 1,4 (8) -p-menthadiene, 1,8-p-menthadiene, p-cymol or N-terpenol. The consumption of sterilization product is 10 kg / ha.

The test results are shown in Table 9.

The same results are obtained with the treatment of the plants in VI. stage of organogenesis.

Table 9

Example number Number of grains laty u isolate Percent sterility Number of grains lat při free pollination Percent deployment grains at free pollination Control 124.9 0.0 145.4 100.0 34 0.0 100.0 138.9 95.5 35 0.0 100.0 89.1 62.3 36 0.0 100.0 137.3 89.6 37 0.0 100.0 109.0  71.1 38 0.0 100.0 107.3 70.0 39 0.0 100.0 128.5 86.0

Example 4

Spring wheat plants of Botanicheskaya 4 are similar to Example 1 in VI. stage of organogenesis treats with 1% aqueous emulsion a mixture of the following sterilizing agents:

1,4 (8) -p-menthadiene and p-cymol, taken at a 1: 1 ratio.

The consumption of the sterilization mixture is 6 kg / ha.

The results of the tests are shown in Table 10. Similar results are obtained in the treatment of plants in stage V of organogenesis.

Table 10

Treatment plant Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 28.9 0.0 30.9 100.0 Mixture ste- rilizač- financial resources 0.0 100.0 27.4 88.7

Example 41

Plants treat genesis in a 1: 1 ratio.

spring wheat of the family Family, similar to Example 1 in VI. stage of the organo1¾ aqueous emulsion of a mixture of sterilizing agents 3-carene and ¢ 6-terpeniol

The consumption of sterilizing agents is 6 kg / ha.

The test results are shown in Table 11.

Similar results are obtained in the treatment of plants in stage V of organogenesis.

Table 11

Treatment plant Number of grains in ear u isolate Percent sterility Number of grains u cob at free pollination Percent deployment grains at free pollination Control 29.5 0.0 35.0 100.0 Mixture ste- rilizač- financial resources 0.0 100.0 29.4 84.0

Examples 42 to 44

Plants of Diploid Rye of Chulpan are treated in VI as in Example 1. stage of organogenesis with 1¾ aqueous emulsion of the following mixtures of sterilizing agents: 1,8-p-menthadiene with 2-pinene (1: 1 ratio), terpenylacetate with p-cymol (1: 1 ratio), 3-carene with terpenylacetate (ratio 1: 1) 1: 1).

The consumption of sterilizing agent mixtures is 6 kg / ha.

The test results are shown in Table 12.

Similar results are obtained in the treatment of plants in stage V of organogenesis.

Table 12

Example number Number of grains in ear u isolate Percent sterility Number of grains in ear at free pollination Percent deployment grains at free pollination Control 44.5 0.0 46.6 100.0 42 0.0 100.0 43.3 92.9 43 0.0 100.0 38.0 81.5 44 0.0 100.0 46.2 99.2

Examples 45 to 48

The Peredovik sunflower plants and Line BK-119 are treated analogously to Example 1 in the V stage of organogenesis with 0.2¾ aqueous emulsion of sterilizing agents h-terpenol or 3-carene.

The consumption of sterilizing agents is 1.2 kg / ha.

The test results are shown in Table 13.

Table 13

Example number Percent deployment nažek při pollination mixtures of pollen in isolate Percent deployment at free pollination Mass 1 000 nažek v 0 Number ster- financial plant in% Content oils nažek in% Keys- vost nažek V -Í Peredovik sunflower Control 85.0 85.0 84.0 0.0 54.7 100.0 45 0.0 80.0 82.3 100.0 53.4 100.0 46 0.0 85.3 84.0 100.0 53.6 100.0 Sunflower Line BK 119 Control 72.6 85.0 60.0 0.0 51.0 100.0 47 0.0 80.4 55.4 100.0 49.4 100.0 48. 0.0 82.5 56.7 100.0 51.7 j 100.0

Claims (2)

SUBJECT OF THE INVENTION A composition for sterilizing plants comprising a diluent, the composition comprising terpenoid compounds having the general formula: A represents -CH-CH "-CHCH" I ^2/2 C _{(CH3) 2} -CH "-CH = _CH-O -, -CH-CH" -CHCH ^{"2 1/2} 'C _{(CH3) 2} C (CH ₃ ) = CH ₂ -CH _O -CH _O -CHCH 2 2,2 C (CH ₃ ) ₂ OH -CH = CHC = CH 1 HC _{(CH3) 2} CH 2 -CH _o -CHCH 2 2 | 2 C _{(CH3) 2 3} 0C0CH -CH ₂ -CH ₂ II ² 'CCCH _{3) 2} or mixtures thereof. 2. The composition of claim 1, wherein the active ingredient is from about 0.1% to about 2% by weight. water as a diluent, characterized in that it is in the form of an aqueous emulsion. that the concentration-