PT89597B - PROCESS FOR THE CREATION OF MICROORGANISMS AND GENETICALLY ANTI-DOTATED MACRORGANISMS AGAINST THE HARMFUL EFFECT OF TIOLCARBAMATES AND CHLOROACETANILIDES ON CULTIVATED PLANTS AND THEIR USE - Google Patents

Abstract

Prepn. of an antidote-carrying Pioneer 3707 maize plant part for lowering the activated plant damage effect of the thiolcarbamates of formula (I) by (a) cultivating maize seals in field soil or under laboratory conditions; (b) applying a cpd. of formula (II) or an acylated deriv. or esterified deriv. to obtain maize plants carrying a herbicidal antidote which is a cpd. of formula (III) to (VIII); (c) harvesting the plants within 30 days of application of (II); (d) isolating the root or shoot, crushing in water and filtering to obtain filtrate and fibrous material; and (e) recovering either the filtrate or fibrous material. In formulae, R1 and R2 are 1-6C alkyl or 3-6C cycloalkyl; or together with a N form a hexamethyleneimino gp.; R3 is 1-5C alkyl or chloroacetanitriles of formula (a); R4 and R5 are H or 1-5C alkyl; and R6 is 1-5C alkyl, 2-5C alkenyl, pyrazolyl alkyl or alkoxyalkyl.

Description

PROCESS FOR THE CREATION OF MICRORGANISMS AND MACRGRGANISMS GENETICALLY BEING

ANTIDOTE AGAINST THE HARMFUL EFFECT OF TIOLCARBAMATES AND OLOROACETANILIDES

IN CULTIVATED PLANTS AND THEIR USE

ESZAKMAGYARORSZÁGI VEGYIMUVEK, Hungarian, industrial and commercial, based in 3792 Sajóbábony, Hungary

RESUME:

The invention relates to a process for the creation and use of microorganisms and macrorganisms carrying an antidote, which are useful in reducing the harmful effect caused in plants cultivated by thiolcarbamates and chloroacetanilides, which comprises the treatment of microorganisms or macroorganisms natural or artificially genetically modified, living in the soil or on the surface of the soil, using the compounds of the general formulas (I) to (I) (VII)

RRR '

Ç

P σ

the s

ll o

| O

- R,

- RR ^L \

R,

R ^L \

R,

Rn

R, (II) (III)

RR - C - S - glutathione

CH _O - CH - COOH ² 1

NH „

CH ₂ - CH - COOH

NH - C - CH, ll (IV) (V) (VI)

COOH

C - S ll o

CH _O - CH - COOH ² l

O - C - CH, (VII)

COOH and the use of living organisms, thus transformed into antidote carriers, in the treatment of agricultural soils, in the natural state or after preservation by lyophilization or by a carrier, in its original state or after elaboration by grinding, spraying, extraction or drying , before, simultaneously with or after the treatment with berbici das, at any time until the germination of the seeds, from the cu

the general formula (VIII), the selectivity of thiolcarbamageral (I) and chloroacetanilides of formula

(VIII) in relation to cultivated plants, preferably corn.

The present invention relates to a process for the creation and use of microorganisms and macrorganisms carrying antidote, which are used against the harmful effect in cultivated plants of the thiolcarbamates of general formula (I), ^E l \ ¹¹ / N - C - S - R;

R '(I) in which

R1 and R2 are the same or different and represent an alkyl group in ol and a cycloalkyl group in 0-6 ' ^{or R} 1 ^{and R} 2 | together with the nitrogen atom, to which they are attached, they form a hexamethylenimino group; and R ^ means an alkyl group in, or by chloroacet

nilides of general formula (VIII),

(VIII) in which

R4 and R4 are the same or different and represent hydrogen or a C1-4 alkyl group; and

Rg is an alkyl group C 2-5 alkenyl '(L alkoxy C ^ _ ^) - (alkyl of C - ^ _ ^) pirazolil- group ^or an (alk :. it into Cj-Cj) ·

It is generally known in plant protection practice that thiolcarbamates of general formula (I), in particular EPTC [S-ethyl-N-thiolcarbamate, N-di (n-propyl)], buylate [S-thiolcarbamate] ethyl-N, Nudi (isobutyl), vernola to [S-propyl-N, N-di (n-propyl) thiolcarbamate], ethylate (S-ethyl-N, N-diethyl thiolcarbamate), (thiolcarbamate thiolcarbamate) of S-ethyl-N-ethyl-N-cyclohexyl), as well as chlorine acetanilides of the general formula (VIII), especially alachlor (2-chloro-2 ', 6'-diethyl-N-methoxy-methyl-acetanilide), acetochlor (2-chloro-2'-ethyl-6'-methyl-N-ethoxy-methyl-acetanilide and propachlor (2-chloro-N-isopropyl-acetanilide) are very effective herbicides.

However, a common disadvantage for both classes of these herbicidally active compounds is their relatively low selectivity against some cultivated plants, in particular maize.

β

When used in the usual herbicidally active doses, mainly EPTC, vernolate and chloride can cause important phytotoxic symptoms. About twenty-five years ago Hoffmann recognized that there are compounds, such as anhydride derivatives of 1,8-naphthalic acid, that have the ability to reduce the phytotoxic effect. Since that time, research has been initiated that was largely developed in order to find synthetic compounds that combat the effect of herbicides that is harmful to cultivated plants.

The antidote action of some classes of compounds has been revealed in several Hungarian patent descriptions mentioned below.

NS. 165 · 756: - antidote action against thiolcarbamates by dichloroacetamide derivatives, for example N, N-diallyl-2,2-dichloroacetamide (code no. R-25 788);

NS. 168-977: ^- Antidotes of the oxazolidine and thiazolidine type, especially advantageous against thiolcarbamates;

NS. 170.214: - Antidotes of the type of sulfone and thiazolidine sulfoxide, useful against thiolcarbamates and chloroacetanilides;

NS. 173 · 775: ^- Sulfide-type antidotes, useful against thiolcarbamates;

NS. 176,867: - Ozazolidine derivatives advantageous to combat thiolcarbamates as antidotes;

NS. 179 · 643: - N-benzoyl.L sulfonyl carbamate compounds, useful as antidotes to thiolcarbamates;

NS. 180.069: - N-benzoylsulfonyl-thiol carbamate compounds, useful against thiolcarbamates, as antidotes; 5

NS. 180,068: - Derivatives of 2,3-dibromopropionami-

- 6 da, useful as antidotes against thiolcarbamates;

We. 174.487, 176 * 784, 180.422, 181.621, 183-997, 185 * 400 and 188.135: - Substitutes of substituted acetamide type advantageous against thiolcarbamates and chloroacetanilides.

Derivatives of dicarboxylic acid with different chemical structures that fight, as antidotes, thiolcar bamates and / or chloroacetanilides have been described in the Hungarian patent specification Nos. 176,669, 176 * 796, 178,892 and 178,895; The effect, as an antidote, of naphthalene carboxylic acid derivatives was revealed in the descriptions of the Hungarian patents numbers 178,883, 179 * 092 and

179 * 093 *

A result of the increased selectivity of thiol carbamates was described in a publication by JE Oasida and others, entitled Thiolçarbamate sulfoxide herbicides, from 1974. The main point of this publication is the fact that EPTC sulfoxide, which is a product from the decomposition of EPTC, it has an advantageous herbicidal action, although it is not harmless in relation to maize. However, the finding of these authors extended only to the use of EPTC sulfoxide itself, which cannot be used for the practical protection of plants, because of its other disadvantageous properties (for example, it cannot be applied to all types of soils).

Despite the results of research described in a large number of patent descriptions, it has not yet been possible to discover a compound that is capable of providing general protection against the harmful effects of the various herbicides, a fact that is also confirmed by the fact that research, oriented towards 0 clarification about them. action of the antidote, failed to develop a single starting point as to the decision on biochemical processes.

occurring in the case of antidotes as additives, or four to the definition of the influence of the mentioned antidotes on the processes induced by herbicides in cultivated plants.

In the course of our research, aimed at reducing the harmful effect on plants cultivated by tiolcarbamate type herbicides of general formula (I) and chloroacetanilides of general formula (VIII), it was found that compounds of general formulas (II) to (VII):

R,

R,

N

(II) ^Rl \ r ₂ o

II s

II

- R:

(III)

glutathione (IV) r ₂ o

CH ₂

CH - COOH

I nh ₂ (V)

CH _O - CH ² I NH

COOH (VI)

C - CH _O - COOH

II ²

CH _O - CH - COOH (VII) ² I

- C - CH _O - COOH

I ² in which, in the compounds of the general formulas (I) to (VIII):

R ^ and R £ are the same or different, and represent a C ^ -C ^ alkyl group or a 0 ^ -0θ, οι cycloalkyl group. R2 and R2 together with the nitrogen atom form a hexamethylenimino group;

means an alkyl group in

R ^ and R ^ are the same or different, and represent hydrogen or an alkyl group in and

R _c represents a G - ^ - C ^ alkyl group, a C2-C5 alkenyl group, a (C - ^ - C ^ alkoxy) - (G - ^ - C ^ alkyl) group or a (alkyl) group in C ^ -C ^) - pyrazolyl formed in the soil or in microorganisms or macrorganisms previously treated with the thiolcarhamates of general formula (I) or their esterified or acylated derivatives can be successfully used to protect against the phytotoxicity of herbicides from both types, when the compounds of general formulas (I) to (VII) are applied to the soil before, or simultaneously with, or after treatment with compositions that are active as herbicides.

re

In this way, the present invention relates to the set of processes, in which natural or genetically artificially manipulated microorganisms or macroranisms that live in soils or on the surface of soils, or whether organs or cultures of cells or cellular components are treated with the compounds of general formulas (I) to (VII), preferably with EPTC.

us

In the course of the treatment process, the decomposition products formed from the compounds of general formulas (I) to (VII), which can preferably be one of the codons of general formulas (II) to (VII) or their mixture or a derivative thereof, esterified or acylated, have an antidote action to the herbicide and are intended for microorganisms or macroorganisms, so that, as a final result, living organisms with antidote are obtained.

The present invention also relates to the use of microorganisms and macrorganisms carrying an antidote, obtained by applying the above processes, to reduce the harmful effect on cultivated plants, by the thiolcarbamates of formula (I) and by the chloroacetanilides of formula (VIII ).

These microorganisms and macrorganisms are used, either in their natural state, or after being preserved by lyophilization or by applying them to a vehicle substance, in its original form or after being processed by grinding, spraying, extraction, drying or in any other way, before or simultaneously with (in the form of a combination produced or tank mix) or after the treatment with the herbicides, at any time until the germination of the cultivated plants, to treat the soil of the fields, by whereas the selectivity of the herbicidally active compounds of the general formulas (I) to (VIII) is increased against cultivated plants, preferably in the case of maize.

The invention is illustrated in detail by the following non-limiting examples.

Example 1

Investigation of the soil, previously treated with EPTO belonging to the herbicides of general formula (I), as a vehicle substance of metabolic products of microbial origin

na, with antidote action.

A field soil, having a ρΞ-KCl value of 5.7, by weight of 46 AE and containing 0.03% of total salts, 1.9% of humus, 14.8 ppm of NO ^ + NO2 » 755 ppm of '^ 53 ppm of ϊ2θϊ 246 ppm of Mg, 60 ppm of Na, 2.1 ppm of Zn, 3.2 ppm of Cu and 151 ppm of Mn was previously treated with a dose of 8 liters / ha of Witox 72 EC, which contains 72% of active substance as an EPTC herbicide formulation, or with a dose of 8 liters / ha of Alirox 80 EG, which contains 72% EPTC and 8% AD-67 £ N- (2, 2-dichloroacetyl) -1-oxa-4-azabicyclo [4,5] decane] as an antidote.

Each 600 g of this soil was weighed in culture pots and 10 grains of different species of hybrid corn were sown, as well as 20 grains of sorghum, each of them as weed.

On the 112th day after sowing, the plants were grown under H MIE 400 / D, z ~ δ daylight bulbs with an illumination period of 16 hours and were watered daily until the water was able to absorb the soil, the green mass of the plants, as well as the length of the corn sprouts were measured, and the external appearance of the corn was evaluated according to the following grading scale:

4: strong deformation, torsion;

3: medium deformation, torsion;

2: mild phytotoxicity;

1: plant intact, healthy.

Then, on the 142nd day after sowing, the piartas were again sown in the culture pots and the soil was treated with a dose of 8 liters / ha or 20 liters / ha, respectively, of Witox 72 EC which contained 72 % of EPTC.

Eleven days after the second sowing, the green mass of the plants and the length of the corn sprouts were measured.

From the data shown in Table I, it can be said that the various species of hybrid maize, sown in the soil previously treated with EPTC or EPTC bearing the antidote AL-67, respectively, tolerated the two doses of 80 liters / ha, respectively. and 20 liters / ha of EPTC, while the sorghum sown as weed has been destroyed.

Table I

Development of the shoot length, the green mash and the outside of hybrid maize, as well as changes in the green mass of sorghum under the effect of treatments with EPTC.

Control No Dose of EPTC treated 8 1 / ha 20 1 / ha

Corn sapling length (cm / plant)

there. sowing 21 8 - 2nd. sowing 19 19 19 Green corn pasta (g / plant) there. sowing 0.56 0.49 - 2nd. sowing 0.5 0.59 0.63

Graduation (corn)

there. sowing 2.0 1.2 - 2nd. sowing 2.0 2.0 2.0

Table I (Continued)

Control No Dose of EPTC treated 8 1 / ha 20 1 / ha

Green mass of sorghum (g / pot) wool. sowing 0.15

2nd. sowing 0.14

Example 2

An experiment was carried out in a laboratory, in a pot culture in order to investigate the synergistic, antagonistic action exerted by different herbicides of the thiolcarbamate type and bacteria carrying an antidote (suspension of bacteria NI-86/21 EPTC, one on the other.

1. - Materials, equipment and processes

1.1 Preparation of NI-86/21 bacteria suspension

A pure strain in culture of NI-86 / / 21 bacteria that decompose EPTC, isolated from the soil, was inoculated in 5 ml of a nutrient element that contains the following components:

Meat extract 1 S Yeast 2 g Peptone 5 g NaCl 5 g Agar 20 g Distilled water 1000 ml PH 7j ⁴

Next, a culture that was more than 5 or 4 days old, but less than 120 days old was used.

Subsequently, the bacteria propagated in the test tube were washed with 10 ml of sterile distilled water. Thus, the suspension contained, in each 1 ml, cer *

of 10 bacteria.

Each 200 ml of physiological nutrient salt solution was poured into 500 ml Erlenmeyer flasks. The composition of the nutrient salt solution as calculated for 1000 ml was as follows:

nh ₄ at ₃

KH ₂ P0 ₄

K ₂ HP0 ₄

NaCl

MgS0 ₄ .2H ₂ 0

CaCl ₂ .2H ₂ 0

GuS0 ₄ .5H ₂ 0

FeS0 ₄ .7H ₂ 0

ZnCl ₂

0.5 g 0.4 g 1.6 g 0.1 g 0.2 g 0.5 mg 0.5 mg 0.5 mg 0.5 mg EPTC active ingredient was weighed into this solution so as to obtain a concentration of 250 ppm. Then, each 2 ml of this bacterial suspension was mixed in the containers which were then shaken in a stirring equipment for 24 hours, to obtain the antidote-bearing bacteria.

Subsequently, the selectivity of the herbicides (EPTC, butylate, vermolate, molinate) introduced in the experiments was observed in themselves and, on the other hand, in their combinations, formed with 10 ml of NI 76/21, suspension

of bacteria carrying an EPTC 250 antidote according to treatments.

Experiment in culture vessels

Plastic containers coated with a sheet of PVC with a surface of 0.8 dm were used, which were able to receive 800 g of soil each. First, 400 g of air-dried soil soil weighing into the pots with a humus content of 1.95% i a pH value of 5 »7 θ a density (according to Arany) of 4. 10 Pioneer 5707 hybrid maize grains, AlftSldi 1 cultivated sorghum grains and 0.5 g of red millet were sown in each pot. The seeds were covered with 100 g of soil in each pot, then EPTC, vernolate and molinate were applied, as active ingredients, in a dose of 6 liters / ha, and butylate as active ingredient in a dose of 24 liters / ha, as manufactured EMV compositions (Che mical Works of North-tungary). The treated soil surface was covered with 100 g of soil in each pot, and then 10 ml of bio-preparation NI-86/21 EPTC was applied to the soil surface according to the treatment list mentioned below:

List of treatments:

untreated control

EPTC 6 liters / ha

EPTC 6 liters / ha + NI 86/21 EPTC 250 10 ml 5 butylate 24 liters / ha butylate 24 liters / ha + NI-86/21 vernolate 6 liters / ha vernolate 6 liters / ha + NI-86/21 molinate 6 liters /there is

EPTC 250 10 ml

EPTC 250 10 ml

molinate 6 liters / ha + NI-86/21 EPTC 250 10 ml

Results of pot culture experiment

The results of the pot culture experiment are shown in Tables II, III and IV.

In Table II it can be seen that the length of the untreated control maize shoots was 22 cm. After the application of EPTC, vernolate or molinate, in a dose of 6 liters / ha, or butylate in a dose of 24 liters / ha, each separately, the shoot lengths were reduced significantly (to 9, 11, 15 or 12 cm, respectively). On the contrary, when 10 ml of bio-preparation Ni-86/21 EPTC 250 were added, in each case, to the herbicides mentioned above, then the effect of reducing the length of the shoots of EPTC, butylate and vernolate (21 , 25 and 21 cm, respectively), while the vernolate phytotoxicity remained unchanged (17 cm).

The data shown in Table III regarding the change in the green mass of corn presented a similar aspect to that of the shoot length.

Corn was completely protected against the effects of the damage caused by EPTC, butylate and vernolate, active as herbicides, using a suspension of NI-86/21 EPTC in a dose of 10 ml in each culture vessel.

This fact is very illustrative in Table IV too, by which the excellent action as an antidote for this biopreparation can be justified based on the external aspect of corn.

red millet and sea bream sown as weeds were described by all treatments, that is, these plants were not protected by the bio-preparation of NI-86/21 EPTC 250.

Table II

Decreased phytotoxic effect caused by EPTC, butylate, vernola to and molinate herbicides in corn sprouts, due to the bio-preparation of NI-86/21 EPTC 250:

Herbicide Dose liters / ha

Shoot length (cm / plant)

Biopreparation of

NI-86/21 EPTC 250

EPTC 6 9 21 Butylate 24 12 23 Vernolato 6 11 21 Molinato 6 13 17 The length of control untreated er: of 22 cm. Table III Action on dough green of corn by herbicides EPTC, butylate, vernolate and molinate, and their combines tions with the bio-preparation of NI-86/21 EPTC 250, respectively mind Herbicide Mass Dose green of corn (10 ~ 2 g / pianta) liters / ha biopreparation of 1 1 H £ 5 1 36/21 EPTC 250 EPTC 6 51 86 Butylate 24 60 83 Vernolato 6 54 88 Molinato 6 57 64

The green mass of untreated control corn was 82 x 102 g / plant.

Table IV

Action on the exterior of corn by the herbicides of EPTC, butylate, vernolate and molinate, and their combinations with the bio-preparation of NI-86/21 EPTC 250 respectively:

Herbicide Dose liters / ha Corn abroad (%) Biopreparation of NI-86/21 EPTC 250 EPTC 6 55 100 Butylate 24 61 100 Vernolato 6 65 100 Molinato 6 58 61

outside of untreated control corn was referred to as 100%.

Rating scale:

100% 95 to 87% 86 to 75% 75 to 51% 50%

intact corn with normal development, mild phytotoxicity, with sapling deformation medium deformation of the saplings, with strong twisting deformation of the saplings, with destruction the maize was described, remained on the ear of the sapling.

Example 5

The action on the selectivity of the

EPTC for the various amounts of the NI-86/21 EPTC 250 bacteria suspension with antidote, prepared as described in example 2.

In this experiment, plastic culture vessels lined with PVC foil, with a surface area of 0.8 dm, were used to accommodate 800 g of air-dried soil in each vessel. First, 400 g of soil was weighed for each pot.

top layer field soil had an average density, a humus content of 1.95% θ pH value of 5.7- 10 grains of hybrid Pioneer 3707 corn were sown as a test plant, as well as 20 grains of sorghum and 0.5 g of millet in each pot. The seed was covered with 100 g of soil, in each pot, then the treatments were carried out with the thiolcarbamates and with the biopreparations of NI-86/21 EPTC 250, respectively, as follows:

Treatments:

untreated control

EPTC 6

EPTC 6

EPTC 6

EPTC 6

EPTC 6 liters / ha liters / ha + NI-86/21 EPTC 1 / ha + NI-86/21 EPTC 250 1 / ha + NI-86/21 EPTC 250 1 / ha + NI-86/21 EPTC 250

250

1000 1 / ha 5000 1 / ha

10000 1 / ha 20000 1 / ha

The treated soil surface was repeatedly covered with 100 g of soil in each case.

The experiment was rated at 10 ^. day after the treatment, when the length of the sprouts and the mass of the maize as well as the green mass of sorghum and millet were measured.

The advantages of combinations containing EPTC together with NI-86/21 EPTC 250 are summarized in Table V.

Table V

Action of the bio-preparation of NI-86/21 EPTC 250 on the phytotoxicity of EPTC in a corn crop (data are given as a percentage of the untreated control).

Treatment

Corn

Mass Green shoot length%%

Green Pasta Green Pasta dc · from Sorghum millet oZ oL / 0 / o

1 EPTC 6 1 / ha 74 41 0 0 2 EPTC 6 1 / ha + NI-86/21 EPTC 250 1000 1 / ha 117 115 0 0 3 EPTC 6 1 / ha + NI-86/21 EPTC 25O 5000 1 / ha 113 112 0 0 4 EPTC 6 1 / ha + NI-86/1 EPTC 25O 10000 1 / ha 95 113 0 0 5 EPTC 6 1 / ha + NI-86/21 EPTC 25O 20000 1 / ha 118 113 0 0

Example 4

An experiment was carried out in a culture vessel for the purpose of comparing the action as an antidote to the biophilized forms of the NI-86/21 bacteria, with the bacteria carrying the NI-86/21 EPTC 250 antidote, against the EPTC, butylate, vernolate and acetochlor, respectively.

1. Material, equipment and method.

1.1- Preparation of lyophilized samples NI-86/21 and NI 86/21 EPTC 250.

A pure NI-86/21 culture was inaccurate. in a Nutrient composition (for composition, see example

2) in 5 test cubes. After cultivating the bacteria. for 3 days, at room temperature, the bacteria mass was washed in 5 ml of a physiological solution (see example 2 regarding the composition of this solution). Each 0.1 ml of the bacteria suspension was thus spread over 100 Petri dishes, for nutrient dishes. After an incubation at 25 ° C for 4 days, the bacterial colonies were suspended in 3 ml of saline each, which were then taken ten to ten for ten Erlenmeyer flasks with 100 ml volume, in order to obtain 10 times 30 ml of bacteria suspension. Of these bacteria, 3 times 30 ml were treated with 3 times 8 of EPTC, then the samples thus obtained were kept at -10 C, until freeze-drying carried out on the 5th ^and · day after that moment.

Lyophilization was carried out in a Labor MIM device, at -40 ° C, without the use of a protective colloid.

In practice, the lyophilized products NI-86/21 and NI-86/21 EPTC 250 were again transformed into suspensions in 60 ml of tap water, each dose in a container, then immediately applied simultaneously with the herbicide treatments, in pre -emergence or pre-sowing.

1.2 - Experiment in culture vessel

In this experiment, plastic pots with PVC sheet lining were used, having a surface of 0.8 dm, which were used to contain 800 g of soil each. First, 400 g of field soil, from the top layer and air-dried, were weighed into said pots. The soil contained 1.95% humus, had a pH value of 5.7 θ and a density (according to Arany) of 46.

> r

In this soil, 10 grains of Pioneer 3901 corn were sown in each pot, as well as 20 grains of HIBAE sorghum. The seed was covered with 100 g of soil, in each case, followed by EPTC in a dose of 6 liters / ha, butylate in a dose of 12 liters / ha, vernolate in a dose of 6 liters / ha and acetochlor in a dose of 3 liters / ha, respectively.

On the surface of the soil so treated, doses of 0.05 x 10, 0.5 x 10,

3

1.0 x 10 and 2.5 x 10 liters / ha, respectively, of the suspension are obtained by lyophilizing the suspension of the NI-86/21 bacteria, as well as the same doses of the NI-86 / / 21 EPTC 250 biopreparation, and , finally the treated soil surface mouth. repeatedly covered with 100 g of soil in each case. Treatments:

Butylate

EPTC 6 1 / ha

EPTC 6 1 / ha + NI-86/21 0.050.5 ~ 1.0-2.5x10 ⁵ 1 / ha EPTC 6 1 / ha + NI-86/21 EPTC 250 0.05 "0.5 ~ 1.0 -2.5xl0 ⁵ 1 Butylate 12 1 / ha

Butylate 12 1 / ha + NI-86/21 0.05-0.5-1.0-2.5xL ³ 1 / ha

1 / ha + NI-86/21 EPTC 250 0.05-0.5-1.0-2.5x x 10 ⁵ 1 / ha z'ha

Vernolate 6 1 / ha

Vernolate 6 1 / ha + NI-86/21 0.05-0.5-1.0-2.5x10 ^ 1 / ha

Vernolate 6 1 / ha + NI-86/21 EPTC 250 0.05-0, 5-1.0-2.5x x 10 ⁵ 1 / ha

Acetochlor 5 1 / ha

Acetochlor 5 1 / ha + NI-86/21 0.05-0.5-1, O-2.5x10 ^ 1 / ha

Acetochlor 5 1 / ha + NI-86/21 EPTC 250 0.05-0.5-1.0-2.5x χ 10 ⁵ 1 / ha

Each treatment was carried out with five repetitions.

The plants were grown until the 102nd day after sowing, or after treatment under HgMIF / 400 W daylight bulbs, using a light intensity of 15 KLX, with a lighting period of 16 hours. The water was supplied daily, in excess of the water capacity. The green masses of the plants and the length of the corn sprouts were measured. The degree of corn torsion was evaluated using the scale described in example 1.

2. - Results and conclusions

The results are shown in Tables VI to XIII. From these Tables it can be said that the lyophilization prepared from the NI-86/21 bacteria, grown in the composition containing 250 ppm of EPTC, reveals a significant antidote action. The phytotoxic effect caused in maize by a dose of 6 1 / ha of EPTC or a dose of 12 1 / ha of butylate or 6 1 / ha of vernolate or 5 1 / ha of acetochlor, respectively, is eliminated by a dose 500, 1,000 or 2,500 1 / ha of NI-86/21 EPTC 250 lyophilisate, prepared in accordance with the present invention. Without pre-treatment with EPTC, the NI-86/21 bacterium has no antidote action.

Table VI

The antidote action of increasing doses of the suspension of the bacterium NI-86/21, against a dose of 6 1 / ha of the herbicide EPTC in corn.

Dose of EPTC 1 / ha NI-86/21 10 ⁵ 1 / ha Corn Evaluation value Comp. From sprouts cm / plant Green pasta g / vase 6 0 4.4 2.1 4 6 0.05 4.8 2.8 4 6 0.5 5.0 3.4 4 6 1.0 5.3 3.4 4 6 2.5 7.6 3.9 3 Control not treated 18.9 5.0 1 ^SZD % 1.7 0.8

Table VII

The antidote action of increasing doses of the NI-86/21 EPTC 250 biopreparation against a dose of 6 1 / ha of the herbicide EPTC in corn.

Dose of EPTC 1 / ha NI-86/21 EPTC 250 10 ⁵ 1 / ha Corn Comp. From sprouts cm / plant Green pasta g / vase Value of evaluation 6 0 4.4 2.1 4

lA-fc / A Table VII (Continued) Dose of NI-86/21 Corn EPTC 1 / ha 10 ⁵ 1 / ha Comp. Green mass of sprouts g / pot cm / plant Evaluation value 6 0.05 5.9 3.1 3 6 o, 5 18.6 5.1 1 6 1.0 20.9 5.7 1 6 2.5 22.5 5.8 1 Control treated no 18.9 5.0 ^SZD % 1.7 0.8 Table VIII pension from herbicide The action as an antidote for NI-86/21 bacteria, against butylate in corn. increasing doses of a dose of 12 1 / ha of Dose of NI-86/21 Corn butylate 1 / ha 10 ⁵ 1 / ha Comp. Green pasta sprouts g / pot cm / plant Evaluation value 12 0 16.1 4.6 3 12 0.05 16.3 4.6 3 12 0v5 17.4 4.8 2 12 1.0 16.7 4.6 2

Table VIII (Continued)

Dose of butylate 1 / ha NI-86/21 10 ³ 1 / ha Corn Value of liation ava- Comp. From sprouts cm / plant Green pasta g / vase 12 2.5 15.5 4.1 2 Control not treated 18.9 5.0 ^SZD 5% 1.7 0.8 Table IX Action as an antidote increasing doses of bio- preparation NI NI-86/21 EPTC 25O against one dose 12 1 / ha of herbicide butylate in corn. Dose of NI-86/21 Corn butylate EPTC 250 C omp. From Green pasta Value of ava- 1 / ha 10 ³ 1 / ha sprouts g / vase liation cm / plant 12 0 16.1 5.6 3 12 0.05 16.0 5.0 2 12 0.5 18.1 5.1 2 12 1.0 18.6 5.0 1 12 2.5 18.7 5.0 1 Control not treated 18.9 5.0 1 ^SZD 5% 1.7 0.8

Table X j The action as an antidote to increasing doses of the NI-86/21 bacterium suspension against a dose of 6 l / ha of the vernolate herbicide in corn:

Dose of vernolate 1 / ha NI-86/21 IO ⁵ 1 / ha Corn Comp. From sprouts cm / plant Green pasta g / vase Avé value liation 6 0 7.5 4.1 4 6 0.05 6.5 3.7 4 6 0.5 8.1 4.0 4 6 1.0 7.7 4.1 4 6 2.5 12.2 4.6 5 Control not treated 18.9 5.0 1 SZD ^ z 77 ° 1.7 0.8 Table XI THE action as antidote to increasing doses of bio- preparation NI-86/21 EPTC 250, against a dose of 6 1 / ha of the vernolate herbicide in corn Dose of NI-86/21 Corn vernolate EPTC 250 C omp. From Green pasta Value of 1 / ha 10 ⁵ 1 / ha sprouts g / vase evaluation cm / plant

Table XI (Continued)

Dose of vernolate 1 / ha NI-86/21 EPTC 250 IO ⁵ 1 / ha Comp. From sprouts cm / plant Green pasta g / vase Value of evaluation 6 0 7.5 4.1 4 6 0.05 8.8 4.3 4 6 0.5 16.6 4.6 3 6 1.0 18.7 5.1 2 6 2.5 19.0 5.0 1 Control not treated 18.9 5.0 1 ^SZD % 1.7 0.8 Table XII Action as an antidote increasing doses of sus- pension from bacteria NI-86/21, against one dose 3 1 / ha of herbicide of acetochlor in corn Dose of NI-86/21 Corn acetochlor 10 ⁵ 1 / ha Comp. From Green pasta Value of 1 / ha sprouts g / vase evaluation cm / plant 3 0 16.0 5.9 3 3 0.05 16.4 4.1 3 3 0.5 16.6 4.8 2 3 1.0 17.3 4.8 2

Table XII (Continued)

Dose of Acetochlor 1 / ha NI-86/21 10 ⁵ 1 / ha Corn Comp. From sprouts cm / plant Green pasta g / vase Value of evaluation 3 2.5 17.9 4.7 2 Control not treated 18.9 5.0 1 ^SZD % 1.7 1 1 (00 ** 0 Table XIII THE preparation action like the NI-86/21 antidote to increasing doses of bio- EPTC 25O, against a dose of 5 1 / ha

j of the acetochlor herbicide in corn:

Dose of acetochlor 1 / ha NI-86/21 EPTC 25O 10 ⁵ 1 / ha Corn Comp. From sprouts cm / plant Green pasta g / vase Value of evaluation 3 0 16.0 3.9 3 3 0.05 18.9 5.1 2 3 0.5 19.1 5.4 1 3 1.0 20.8 5.5 1 3 2.5 21.4 6.0 1 Control not treated 18.9 5.0 ^SZD % 1.7 0.8

Example 5

A laboratory experiment was carried out in culture vessels in order to investigate the antagonistic action of EPTC in a corn culture, aqueous filtrates and acetone filtrates, as well as the residual fibrous substances obtained from the sprout and root of small maize plants, previously treated with EPTC.

1. - Materials, equipment and method

1.1. - Preparation of an antidote-bearing corn

Plastic, culture vessels, lined with a PVC sheet, each with a surface of 4 dm, were used, which were adequate to receive about 6000 g of dry air in each container. First, 4000 g of field soil with a 1.95% humus content, a pH value of 5.7 θ and a density of (according to Arany) of 46 were weighed into the containers. 50 Pioneer 3707 corn kernels are placed in each container, and cc is filled with 1000 g of soil in each case. Subsequently, the culture vessels, designated (Ε), were treated with a dose of 6 1 / ha of EPTC. The treated soil surface was repeatedly covered with 1000 g of soil in each container.

The corn plants, which received the treatment (I were grown, from the treatment until the 142nd day, under HgMIF / 400 W daylight bulbs, with a light intensity of 11 KLX, in a daily lighting period of 16 hours, at a temperature of 26+ 4 ° C.

The plants were daily sprayed from above to water capacity. In 142. cLia, the sprouts of the piers were cut separately in each container and the roots were washed to remove the soil.

1.2 - Separation of filtrates and fibrous substances from corn

1.2.1. - The processing of corn sprouts previously treated with EPTC (E)

After cutting, 55 g of the seedling (Ha) of the corn previously treated with EPTC (E) were immediately ground in 500 ml of water in a swirling machine. The grinding was divided into two halves, 250 ml were filtered through a laboratory glass filter and, in this way, an aqueous filtrate (Sz) was obtained, which was marked with the code E-Ha-Vi-Sz.

fibrous material remaining in the filter was shaken with 50 ml of acetone in a stirring tunnel and the acetone phase was removed. The acetone phase was labeled with the code E-Ha-Ac-Sz.

The remaining 250 ml aqueous milling portion was filtered through a vacuum filter and the fibrous substance remaining on the filter was removed as a prepared sample labeled with the code E-Ha-Vi-R.

1.2.2 - Processing of the corn root previously treated with EPTC processing was carried out in a manner similar to that described above, with the exception that 28 g of the roots crushed with a knife were treated with 400 g of tap water in a swirling machine. The aqueous filtrate and then the acetone filtrate were obtained from 200 ml of the mill; the fibrous substance was obtained from the other part with a volume of 200 ml.

These fractions were cross-linked with the following codes:

E-GyU-Vi-Sz

E-GytJ-Vi-R

E-Gyb-Ac-Sz.

1.5 - Investigation in a culture vessel on the EPTC antagonistic action of filtrates and fibers obtained from corn.

1. - Sequence of experience

Plastic pots with PVC sheet lining were used, having a surface of 0.8 dm each, which was adequate to contain 800 g of soil each. First, 400 g of field soil, from the upper layer, were weighed into each of the aforementioned pots, with the properties described in 1.1.

Then, 10 Pioneer 5707 corn kernels were sown in each pot, with sowing covered with 100 g of soil in each pot, then EPTC was applied in a dose of 6 liters / ha. The treated soil surface and the untreated pots were several times covered with 100 g of soil, in each case, then 100 ml of the aqueous filtrates, 100 g of the fibrous material or 5 ml were applied to each pot, respectively. of the acetone filtrates.

The plants were grown under the conditions described in 1.1, from sowing to the 7th day, then bird. the shoot length and the weight of the green mass of the corn, as well as the external aspect of the corn, were used using the evaluation scale described in example 1.

Table XIV

The action of aqueous filtrates (Sz) (Vi) and acetone (Ac), as well as residual fibrous substances (R) obtained from the previously treated corn seedling (Ha) (E) with a dose of 6 liters / ha of EPTC , on the selectivity of

EPTC (6 liters / ha) as a herbicide in corn. Treatment Corn Cornp. of sapling cm / plant Green mass 10 ² g / plant Evaluation value Control not treated 19 72 2.0 EPTC 9 56 1.2 EPTC + (E-Ha-Vi-SZ) 19 62 1.8 EPTC + (E-Ha-Vi-R) 19 72 2.0 EPTC + (E-Ha-Ac-Sz) 20 70 2.0 E-Ha-Vi-Sz 20 72 2.0 E-Ha-Vi-R 20 79 2.0 E-Ha-Ac-Sz 18 68 2.0

From the data in Table XIV, it is evident that the phytotoxic effect of a dose of 6 liters / ha of the herbicide EPTC has been completely eliminated by the extracts or by the residual fibrous material obtained from the roots of small corn plants, previously treated with a dose of 6 liters / ha of EPTC. As a result of the antidote action of these biopreparations, the shoot length and the external aspect of the cultivated plant remained intact and healthy, similar to those of the untreated control.

Table XV

Action of aqueous filtrates (Sz) (Vi) and acetone (Ac) as well as residual fibrous substances (R) obtained from previously treated roots (GyB) of corn (E) with a dose of 6 liters / ha of EPTC , on the selectivity of the herbicide EPTC (6 liters / ha) in maize.

Treatment Corn Comp. of sapling cm / plant Green mass IO ² g / plant Evaluation value Control not treated 19 72 2.0 EPTC 9 56 1.2 EPTC + (E-GyB-Vi-Sz) 5 16 1.0 EPTC + (E-Gyo-Vi-R) 19 72 2.0 EPTC + (E-GyB-Ac-Sz) 18 70 2.0 E-GyB-Vi-Sz 7 22 13 E-GyB-Vi-R 20 73 2.0 E-GyB-Ac-Sz 19 68 2.0

From these data in Table XV, it is quite evident that both the acetone filtrate and the residual fibrous material obtained from the roots of small corn plants previously treated with a dose of 6 liters / ha of EPTC exert an excellent action as antidotes.

Claims (4)

i wool. - Process for the creation of microorganisms [antidote carriers against the harmful effect of thiol; carbamates, and chloroacetanilides, which are useful in j; reduction of the harmful effect in cultivated plants causes [[by the tiolcarbamates of general formula (I) i ' Ε, O ^x \ | N - C - S - R ₃ (I) | in which ! R ^ and R2 are the same or different, and represent an al i group kyl at C ^ _g or a cycloalkyl group at Cj_g, or R ^ and R ~, together with the nitrogen atom, to which they are attached, ¹ form a hexamethylenimino group; and jj Rj means a C1-4 alkyl group, or by chloroacetaniI read from general formula (VIII) i (VIII) in which R ^ and R are the same or different and represent hydrogen or alkyl C ₁ _j; θ Εθ means a C - ^ _ j alkyl, C2-5 'alkenyl ( ^at 2 C ₁ _j coxy) - (C - ^ _ j alkyl) or C ₁ _j pyrazolyl alkyl), - 35 characterized by the fact that it comprises the treatment of natural or genetically artificially manipulated microorganisms that live in the soil of the fields or are prepared in laboratory conditions, propagated and transformed into physiological suspension in an amount between 0.001 and 99 percent by weight, with a of the compounds of the following general formulas (I) to (VII) 0 8 N - C - S - S (I) R, R. Rn R,, N S II o II s ll o - R- R: (II) (III) RR. ) N - C - S - glutathione (IV) RRRn R,: n - o - s CH _O - CH - COOH ² I NHzn: N - G - S - CH _O - OH - COOH « ² I (v) (VI) NH - C - CH ₂ - COOH, Ν R. Ç Í1 o CH ₂ - CH - COOH (VII) O - C - CH ^ - COOH in which R ^, R ₂ and R ^ have the meanings as defined above, either with their acylated or esterified derivatives, or with a mixture of 0.0001 to 99% by weight of ones with others) or with other pesticides, regulators, antidotes, fertilizers or other organic or inorganic substances, natural or synthetic, corresponding to an active ingredient content of 5 to 50 000 g / ha or 0.002 to 5 000 ppm, respectively; and then, if necessary, preserving the microorganisms bearing the antidote thus obtained by lyophilization or by mixing with an organic or inorganic carrier, and finally, optionally processing the mixture containing the said microorganisms by grinding, spraying extraction or any other method. 2s. Process according to claim 1, characterized in that it comprises the treatment of a class of bacteria registered with the NS. NI-86/21, with those with general formulas (I) to (VII), in order to obtain a suspension of bacteria carrying an antidote against the harmful action of said compounds on the cultures. 3 ^. - Process for the creation of microorganisms that carry an antidote against the harmful effect of thiolcarbamates and chloroacetanilides, which are useful in reducing the harmful effect caused in plantations useful by the tiolcarbamates of general formula (I) Rn ‘N - C - S - R (I) in which Rz R-j_ and ϊ-2 are the same or different, and represent an alkyl group in Cj__g or cycloalkyl at 0 ^ _θ, or R ^ and R2 together with the nitrogen atom, to which they are attached, form a hexamethylenimino group; and R means an alkyl group in or of chloroacetanilines of the general formula (VIII) (VIII) in which R 2 and R 2 are the same or different and represent hydrogen at a C 1-4 alkyl group; ^and R ^ is an alkyl group alkenyl, C2_ ^ '(alkoxy C ₁ _ ^) - (C ^^ alkyl) or pirazolil- (alkyl wcaracterizado by comprising treating a species of natural or artificially genetically macroorganisms preferably handled from plants of higher orders, which live in soils or are grown under laboratory conditions, in a solid or liquid physiological medium, or the respective culture of organs, tissues or cells, with one of the compounds of general formulas (I) to (VII ), in which R,, I ?2 ^and R are as defined above, or with their acylated or esterified derivatives, or with their mixtures of 0.0001 to 99% by mass or with other pesticides, regulators, antidotes, adu bos or other organic or inorganic, natural or synthetic substances, corresponding to an active ingredient content of 5 to 50 000 g / ha or 0.002 to 5 000 ppm, respectively ss i I ι then, if the harvest of the macrorganisms thus transformed into carriers of the antidote is carried out, within the period of | 30 days after treatment and, if necessary, carry out! the preservation and / or processing of the aforementioned macrorganisms by crushing, spraying, extracting, drying or by any other method. 4th. - Process for reducing the harmful effect of plantations caused by thiolcarbamates of general formula (I) or chloroacetanilides of general formula (VIII), in which R ^, R ₂ , R ^, R ^, R ^ θ R ^ are in accordance was defined in vindications 1 or 3, characterized in that it comprises the treatment of agricultural soils with a dose of 5 ^- 5,000 liters / ha of a composition containing the microorganisms or macrorganisms that carry an antidote according to any of the claims 1 to 3 in a content of 0.000 1 to 99% by weight, before sowing or emergence of cultivated plants, before or simultaneously with or after treatment with a composition containing a thiolcarbamate of general formula (I) or a chloroacetanilide of formula general (VI 'at any time until the germination of the seeds of the cultivated plants.