CS273345B2 - Agent for plants' anthers sterilization - Google Patents

Abstract

The solution concerns an agent for sterilization of plant anthers containing solvent and the active substance is represented by terpene compounds of general formula I<IMAGE>,where A means<IMAGE>.The agent is optimally used in form of 0.1 to 2 percent water-based emulsion.

Description

The invention relates to biology and agriculture, and in particular to a method of sterilizing plant pillers for use in selection and seed production.

Today, the world's challenge is to intensify agriculture, in particular to increase the profitability of cereals, feed, vegetables and technical cultures through the widespread use of first generation hybrids. Hybrids differing from parental forms by higher yields (about 25 to 30%) and better product quality can be obtained by crossing.

There is a method for 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 involving the acquisition of sterile analogs, sterility fixators and fertility restorers. Especially promising are methods of sterilizing plant anthers with chemical sterilizers (gametooids). The use of gametocides is considerably more economical than the use of a cytoplasmic pollen sterility system, as there is no need to develop such forms as sterile analogs, analogues for fixation of sterility in female forms and for fertility recovery 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 control of their mass production.

Currently, about 200 compounds have been found to have gametooid activity and their chemical structure belongs to different classes of chemical compounds. Gametooids must cause complete pollen sterility in the treated plants while maintaining the ability of the egg cells to live and ensure a sufficiently high (at least 70% control) testicle formation under free pollination. Their phytotoxicity and toxicity to warm-blooded animals must be minimal.

Methods of sterilizing cereal anthers are known (L. Dzh, Nikell, Rosta rasteni controllers. Primenenie v rusticske chozjajetve, Moscow, et al., Kolos, 1984, p.

- 31; SU, A. 906457), which consist of treating plants with sterilizing agents such as 2-chloroethylphosphonic acid (Etrel), maleic anhydride, di- (polyfluoroalkyl) phosphoric acid and its salts and others. Treatment of plants with sterilizing agents is carried out in V or VI. stage of organogenesis (according to P.M, Kupermann).

The processes of formation and differentiation of flowers begin in the V. atadium of organogenesis. Towards the end of this stage, neoplasms, archesporium tissues are formed. 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. VI. Stage is characterized by the process of formation of the flower (micro and macrosporogenoza). Separate mononuclear pollen grains are formed at this stage (P, M, Kupermann, Morphofiziologija rasteniya, Moscow, 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 in a period between the appearance of the second stem and the ear. As a sterilizing agent, heterocyclic compounds are used, the main representatives of which are 2-carboxy-3,4-methanpyrrolidine or 2-methoxycarbonyl-3,4-methanpyrrolidine. These compounds are used in admixture with diluents and surfactants.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a new sterilizing agent which allows sterilization of plant anthers in a wide range of cultures with a high sterilization degree, while maintaining the high ability of the seeds to deploy in free pollination.

The problem is solved by treating the anthers of the plants in a fifth and / or sixth stage of organogenesis with a sterilizing agent containing a diluent which contains terpene compounds of the formula I as active ingredient.

in which A denotes

OO exo-OCH, exo-OCCFl, exo-QCCCl4, exo-OCCHgCl, exo-OCCHgBr, exo-OCCCH3, exo-OCH ₂ CH ₂ Cl, exo-OCH (CH ₂ Cl) ₂ , exo-OPh , exo-OCCH-j, endo-OCCH-j, or mixtures thereof.

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

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

The composition according to the invention is used as follows:

Plants such as winter wheat and spring wheat, diploid and tetraploid rye, triticale, millet, sunflower and the like are treated with a sterilizing agent according to the invention, in which the terpene compounds of the general formula are used as active ingredient.

wherein A is exo-OCH, exo-OCCF4,

0

II II exo-OCCO1 ^, exo-oCCHgCl, exo-OCCHgBr,

exo-OCH 2 CH ₂ Cl, exo-OCH (CH ₂ Cl) ₂ , exo-OPh, endo-OCCH 2

Or mixtures thereof.

Terpene compounds can be used in conjunction with any suitable diluent. Suitably, water is used as the diluent. A 0.1 to 2% aqueous emulsion of said compounds is preferably 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 are usually added to the ready-to-use solutions. dispersing agents and adhesives.

The sterilizing agent is applied to the plants by a variety of treatments, such as by 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 the VI stage of organogenesis is expedient. The total dose of the sterilizing agent is 0.6 to 20 kg / ha.

All terpene compounds of the invention under consideration as a sterilizing agent were tested for toxicity in animal experiments. The results of the tests showed that said compounds were slightly toxic or practically non-toxic. For example, LD 2 -qu of 2-exo-hornyl monochloroacetate for white mice is 480 mg / kg animal weight, 2-exo- (2-chloroethoxy) hornane is ΙΒ 130 2130 mg / kg animal weight, 2-exo-hornyltrifluoroacetate is 2 700 mg / kg animal weight, BD ^ qu 2-exo-bornyltrichloroacetate 3 040 mg / kg, BD ^ qu 2-exo-phenoxyhornan 3500 mg / kg, BD ₁₀ for 2-exo-homyl formate for rats is 5,000 mg per kg animal weight, for 2-exo-hornyltrimethyl acetate for rats, is 3000 mg / kg animal weight, and u for 2-endo-bornylacetate for rats is 5,000 to 10,000 mg / kg animal weight.

The sterilizing agents according to the invention are obtained by known methods in the reaction of camphene with electrophilic agents (with carboxylic acid halides or carboxylic acid alkyl halides or alcohols) in an acid medium at a temperature of 30 to 100 ° C.

The gametocidal activity of the sterilizing agents of the invention was determined by field experiments in various soil-climatic zones with partial areas of 10 m @ 2, with two, three to four replicates. Each sterilizer has been tested for at least 5 years.

The developmental stage was controlled cytologically. Plant treatment with a sterilizing agent was performed at the beginning of the fifth stage of Kupermann's organogenesis. The type procedure is as follows;

After the ears have been grown, the largest ears and the others are insulated in parchment bags. For wheat and triticale, individual isolates are used. In rye, the ears of 5 to 7 different side-by-side plants are sealed in one common insulator. For millet, each lath is isolated separately. The percent sterility (X) for wheat, rye, triticale and millet is determined by the formula

IN.

and.

CS = 273345 B2 / number of seeds fertilized per isolate of treated plants \ number of seeds fertilized per isolate of untreated control plants

-i6.

The number of grains in the uninsulated spikes of the control plants during free pollination is taken as a comparative, ie. 100% deployment.

To obtain reliable data, 20 to 25 wheat and triticale isolates are used in each experiment, 10 to 15 rye and millet isolates are used in each experiment.

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 mixed pollen collected from 20-25 isolated dressings, and 15 plants were left free to pollinate to control deposition of achenes in pollen pollen from untreated parental plants.

The sterility of the pollen of sunflower plants is judged by the fertility of the pollen and the germination, the morphological peculiarities of the male germ cells and the ability of the seeds to deploy untreated plants after pollination. The viability of the egg cell is determined after seed formation in the treated plants by free male pollination.

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

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

For a better understanding of the invention, the following embodiment variants of the process according to the invention are provided.

Example 1

Mironovskaya 808 winter wheat plants are treated by spraying a 2% aqueous emulsion of 2-exobornyl bromoacetate in a V stage of organogenesis with a portable sprayer.

0.1% wt. 14 ° CI2 ^kalciuniall yl ^{b £} ® ^nzensu l ^{f 'he} ^' ^{t 'u} · 7 ^as adjuvants to the emulsion can be from 0.01% by weight, dimethyl sulfoxide. The consumption of the product is 12 kg / ha. As a control, plants treated with a diluent without sterilizing agent. The test results are shown in Table 1. Analogous results are obtained with the treatment of the plants in VI. stage of organogenesis. ,,

Jli

Examples 2 to 3 ^J

The procedure is analogous to Example 1. A 2% aqueous emulsion of 2-exo-bornyltrifluoroacetate and 2-exo-bornyltrichloroacetate is used as a sterilizing agent. The test results are given in Table 1,

Examples 4 to 7

The process is carried out analogously to the process described in Example 1 except that 0.5% by weight of aqueous emulsion of 2-exo-2-chloroethoxybornane, 2-exo-bornyl monochloroacetate, 2-exo-bornylformate or 2-exo-bornylacetate are used. . The consumption of the product is 3 kg / ha. The test results are given in Table 1.

Examples 8 to 9

The process was carried out analogously to that described in Example 1, except that a 1% aqueous emulsion of 2-exo-phenoxybornane and 2-exo- (2-chloro-1-chloromethylethoxy) was used.

CS'273345 B2

bornanu. The consumption of the product is 6 and 10 kg / ha. The test results are: listed in the table Table 1 Example number number of ear grains in isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 34.5 0.0 40.5 100.0 1 0.0 100.0 38.2 94.3 2 0.0 100.0 28.1 69.4 3 0.0 100.0 28.3 69.9 4 0.5 98.6 32.3 79.7 5 0.5 98.6 31.4 77.5 6 0.2 99.4 33.7 83.2 7 0.0 100.0 31.7 - 78,3 8 0.0 100.0 38.9 96.0 9 0.0 100.0 36.1 89.1

Examples 10 to 16

Plants of spring wheat Moskovskaya 35 were in VI. stage of organogenesis (according to Kupermann) treated with 1% aqueous emulsion of 2-exo-bornyl monochloroacetate, 2-exo-bornyl imonobromoacetate, 2-exo-bornyltrifluoroacetate, 2-exo-bornyl acetate, 2-exo- (2-chloroethoxy) bornane, 2-exo 2-endo-bornyl acetate. The emulsion contains an emulsion of 0.1% by weight. kalciumalkylbenzensulfonátu ^{σ χ2 "σ ΐ4} eájuvans ^and 0.01% by weight, Ν, Ν-dimethylformamide. The consumption of the product is 6 kg / ha. Diluent-treated plants without sterilizing agent serve as a control. The test results are shown in Table 2. Analogous results are obtained in the treatment of plants in the V &apos; stage of organogenesis.

Table 2

Example number Number of ear grains in isolate Percent sterility and Number of ear grains in free pollination Percentage of grain application in free pollination Control 34.9 A - 36.4 100.0 10 0.0 100.0 22.1 60.7 11 0.0 , 100.0 22.7 62.4 12 0.0 100.0 30.4 83.5 13 0.1 99.7 20.9 57.4 14 0.0 100.0 22.6 62.1

Table 2 - continued

Example number Number of ear grains in isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination 15 Dec 0.0 100.0 27.5 75.5 16 0.2 99.4 35.6 97.8

Examples 17 to 23

Plants of diploid rye of Chulpan species in VI. stage of organogenesis (according to Kupermann) treats with 1% aqueous emulsion of 2-exo-bornyl monochloroacetate, 2-exo-bornyl monobromoacetate, 2-exo-bornyltrichloroacetate, 2-exo-phenoxybornane, 2-exo-2-chloro-1-chloromethylethoxybornane or 2 The emulsion contains 0.1 wt. θ] _ι 2 “θ14 of ciumalkylbenzenesulphonate and 0,01% by weight of dodecylsulphate. The consumption of the product is 6 kg / ha. As a control, plants treated with diluents without a sterilizing agent.

The test results are shown in Table 3. Analogous results are obtained in the treatment of plants in stage V of organogenesis.

Table 3

Example number Number of ear grains in isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 49.2 w 54.1 100.0 17 0.7. 98.7 46.2 85.4 18 0.6 98.9 38.6 71.3 19 Dec 0.0 100.0 38.1. 70.4 20 May 0.3 , 99.4 52.3 96.7 21 0.5 99.1 53.5 98.9 22nd 0.0 > 00,0 53.6 99.1 23 0.4 ¹ 99.3 V · 49.0 90.6

Examples 24-26

The Belta tetraploid rye plants are treated with a 1% aqueous emulsion of the following sterilizing agents in the V stage of organogenesis (according to Kupermann): 2-exo-bornyl (trifluoro) acetate, 2-exo-bornyl (trichloro) acetate, 2-exo- fenoxybornane. The consumption of sterilization product is 10 kg / ha.

The results of the tests performed compared to the control and the standard are shown in Table 4.

and;

Table 4

Example number Number of ear grains Percentage Number of ear grains in free pollination Percentage of grain application in free pollination in isolate sterility Control 39.3 0.0 44.2 100.0 24 0.0 100.0 38.1 86.2 25 0.0 100.0 36.0 81.4 26 0.0 100.0 39.2 88.6 Example 27 The Kharkovskaya diploid rye 55 plants are treated analogously to Example 1 in V and VI, the organogenesis stage with 1.0% aqueous 2-exo-phenoxyhornan emulsion. The sterilization consumption is 6 kg / ha. The test results are shown in Table 5.

Table 5

Example number Conceive of grains of ear In Isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 31.5 0.0 39.4 100.0 27 Mar: 0.0 100.0 34.3 87.0

Example 28

Similarly to Example 1, triticale plants of the PRAG-101 species are treated with a 2% aqueous emulsion of 2-exo-phenoxybornane in the V stage of organogenesis. The consumption of sterilizing agent is 12 kg / ha. The test results are shown in Table 6.

Tahulka 6

Example number Number of ear grains in isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 41.3 0.0 45.8 100.0 28 0.3 99.3 39.7 86.7

Example 29

The triticale Amfidiploid 206 plants are treated analogously to Example 1 in stage V of organogenesis with a 2% aqueous emulsion of 2-exo-phenoxyhornan. The consumption of sterilizing agent CS · -273345 B2 is 12 kg / ha. The results of the tests are shown in Table 7.

Table 7

Example number Number of ear grains in isolate Prooento sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 23.4 0.0 25.2 100.0 29 0.2 98.9 25.2 100.0 Examples 30 to 39 Mironovskoye 94 millet plants are treated analogously to Example 1 in the V stage of organogenesis with a 1% aqueous emulsion of the following sterilizing agents: 2-exo-bornyl (trifluoro) acetate, 2-exo-phenoxyboraan, 2-exo- (2-chloro-1) 2-exo-bornyl monochloroacetate, 2-exo-bornyl monobromoacetate, 2-exo-bornyl (trichloro) acetate, 2-exo-bornyl acetate, 2-exo-bornyltrimethyl acetate, 2-exo-bornyltrimethyl acetate, 2-exo-bornyltrimethyl acetate, 2-exo-bornyl acetate, 2- exo - (2-chloroethoxy) bornane or 2-exo-bornyl formate, respectively. Consumption the sterilizing agent is 10 kg / ha. Test results in comparison with control are listed in Table 8. Table 8 Example number Number of lath grains in the isolate Percent sterility Number of grains in free pollination Percentage of grain application in free pollination Control 105.1 0.0 153.3 100.0 30 0.0 100.0 123.6 80.6 31 0.0 100.0 130.0 84.8 32 0.0 - 100.0 90.2 58.8 33 0.0 100.0 101.2 66.0 34 0.0 100.0 105.2 68.6 35 0.0 ^: 100.0 128.5 83.8 36 0.0 100.0 128.4 83.8 37 0.0 100.0 136.9 89.3 38 0.0 100.0 98.7 64.4 39 0.0 - 100.0 135.7 88.5

Examples 40 to 45

Peredovik sunflower plants and Line BK 119 are treated analogously to Example 1 with a 0.2% aqueous emulsion of the following sterilizing agents: 2-exo-bornyl acetate, 2-exo-bornyl formate, 2-exo-phenoxyboman, 2-exo-bornyl mono-chloroacetate, 2 -exo- (2-chloroethoxy) bornane or 2-exo-bornyltrichloroacetate. The treatment is carried out at the V stage of the organogenesis at the asterisk phase. The application rate is 1.2 kg / ha. The test results are shown in Table 9.

Table 9

Example number Percentage of seedings in the isolate when pollinated with pollen mixtures Percentage of free pollination Weight Qty Obeah oils in g Germination nažek in% 1 000 nažek in g steril. plant in% Peredovik sunflower Control 85.0 85.0 84.0 0.0 54.7 100.0 40 0.0 68.3 78.0 100.0 54.5 100.0 41 0.0 85.4 82.7 100.0 51.4 100.0 42 0.0 79.8 89.0 100.0 53.9 100.0 Sunflower Line VK 119 Control 72.6 85.0 60.0 0.0 51.0 100.0 43 0.0 77.4 55.8 100.0 51.4 100.0 44 0.0 88.2 59.5 100.0 50.3 100.0 45 0.0 83.5 56.0 100.0 50.1 100.0

Example 46

Spring wheat plants of the Botanicheskaya 4 type are treated analogously to Example 1 with an aqueous 1% emulsion of a mixture of sterilizing agents such as 2-exo-bornyl monochloroacetate with 2-endo-bornyl acetate, taken at a ratio of 1: 1.

10.

Table 10

Treatment plant Number of ear grains in isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 28.9 0.0 30,9 ‘ 100.0 Sterile mixture beginnings Wednesday 0.0 100.0 29.0 93.8

Examples 47 to 49

Analogous to Example 1, diploid rye plants of &lt; RTI ID = 0.0 &gt; 1 &lt; / RTI &gt; bornyltrimethyl acetate (1: 1 ratio), a mixture of 2-exo-bornyl formate with 2-exo-phenoxy bornane (1: 1 ratio).

The test results are given in Table 11,

Table 11

Example number Number of ear grains in isolate Percent sterility Number of ear grains in free pollination Percentage of grain application in free pollination Control 44.5 0.0 46.6 100.0 47 0.0 100.0 40.5 86.9. 48 0.0 100.0 36.4 78.1 49 0.0 100.0 38.8 83.3

OF THE INVENTION

Claims (1)

OF THE INVENTION 1. A composition for sterilizing anthers of plants comprising a diluent, characterized in that it contains, as an active ingredient, terpene compounds of the general formula I in which A denotes II exo OCH exo OCCHgCl ^ss' ^ oxo-OCCP ^, exo OCCCl ^, exo OCCHgBr, exo OCCÍCH ^ J ^, exo-0CH ₂ CH ₂ 01 exo-0CH (CH ₂ Cl _2), exo -OPh,