RU2787400C1 - Agrobiovit p1 - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to agriculture. The composition for plant protection against apple and/or pear scab contains Trichoderma viride, Azotobacter chroococcum, Bacillus pumilus, Pseudomonas fluorescens. Additionally, the composition may contain Bacillus subtilis and Pythium oligandrum.

EFFECT: proposed composition for plant protection from apple and/or pear scab has high biological activity, reduces damage to plants and plant parts, reduces the loss of harvested fruit caused by scab.

4 cl, 7 tbl, 2 ex

Description

Field of invention.

The present invention relates to compositions for protecting plants from harmful diseases, reducing damage to plants and plant parts, as well as losses in harvested fruits or vegetables caused by such diseases, in particular scab.

State of the art

Apple scab is a harmful plant disease. Exposure to a phytopathogen leads to premature leaf fall, drying and death of shoots, infected ovaries crumble. In this regard, the quantitative and qualitative characteristics of the yield are deteriorating, since the commercial qualities of the fruit are significantly reduced. During storage, scab-damaged apple fruits are susceptible to infection with fruit rot and various molds. Harm from the impact of infection is observed in fruit nurseries, where seedlings are severely inhibited. Under favorable development conditions, this plant disease can destroy the entire crop. In addition, the infection reduces the frost resistance of trees.

Apple scab is a widespread plant disease. It is especially harmful in the North Caucasus, Transcaucasia, on the southern coast of the Crimea, in the Polissya of Ukraine, in Moldova, Belarus, the Baltic countries, and the northwestern regions of the Russian Federation. In some years, outbreaks of the disease are observed in the central regions of Russia, forest-steppe Ukraine.

Apple scab is caused by a pathogen that has two stages in the development cycle: the teleomorph Venturia inaegualis, the anamorph Fusicladium dendriticum. The phytopathogen is

highly specialized parasite. The infection affects leaves, sepals, fruits, stalks, less often young shoots.

Existing scab control measures include:

1) agrotechnical, such as cleaning and destruction of fallen leaves; autumn plowing of the soil in the garden; digging trunk circles; selection of well-drained and ventilated areas for laying new plantations; cultivation of resistant varieties,

2) chemical, such as preventive and eradicating spraying of trees, fruits, soil, fallen leaves in spring, autumn and during the growing season with alternating systemic and preventive fungicides based on strobilurins, triazoles, inorganic substances, copper compounds and other substances, and

3) biological, in particular spraying during the growing season with biological products.

One of the most effective anti-scab preparations are Delan, VG from Basf and Fitosporin-M, manufactured by BashIncom.

Fitosporin-M is a live spore bacterial culture of Bacillus Subtilis 26 D, which suppresses the reproduction of many fungal and bacterial plant diseases with its products. Effectively protects plants from powdery mildew, helminthosporium root rot, brown rust, rhizoctoniosis, alternariosis, dry and wet rot of tubers, phomosis, mucous bacteriosis, tracheomycosis wilt, peronosporosis (downy mildew), black bacterial spot, bacterial cancer, cercosporosis, rot (fusarium rot, black moldy rot, green penicile rot), storage rot (white rot, gray rot, dry black rot, phomosis, monolial rot), late blight, snow mold, scab, fruit rot, rust, white spot, rusty blotch, pink mold, American powdery mildew, etc.

In addition, Fitosporin-M has powerful anti-stress, growth-promoting, immunostimulating properties, which is important for climatic, chemical, pesticide and other stresses and for increasing crop productivity and reducing re-infections.

Delan is a universal fungicide of contact action, highly effective in the fight against many fungal diseases of fruit crops and grapes.

The active ingredient dithianon affects several enzymes of the fungus at once, as a result of which the emergence of resistance in pathogens to dithianon is practically excluded. Delan is a contact fungicide for preventive use. After treatment, a rain-resistant protective layer is formed on the plant, which effectively inhibits the germination of fungal spores. At the same time, a certain part of the active substance (the so-called "depot") can always dissolve in water, which ensures a long-term and stable fungicidal effect of the drug.

At the same time, the development of new biological preparations with an effective protective effect against scab remains relevant.

Disclosure of invention

The present invention provides compositions and/or combinations of active biological ingredients for protecting crops from scab.

In one embodiment of the present invention, the composition contains Trichoderma viride, Azotobacter chroococcum, Bacillus pumilus, Pseudomonas fluorescens.

In another embodiment, the composition of the invention contains Bacillus subtilis, Bacillus pumilus, Azotobacter chroococcum, Pseudomonas fluorescens, Trichoderma viride, Pythium oligandrum.

Implementation of the invention

Example 1. Evaluation of the effect of preparations Agrobiovit P1 and Agrobiovit P2 on the growth of pure cultures of the apple scab pathogen in laboratory conditions.

The study was carried out on monospore isolates of the apple scab pathogen Venturia inaequalis (Cook) Wint. from the collection of phytopathogenic micromycetes of the laboratory of biotechnological control of phytopathogens and phytophages. 8 different isolates were analyzed: 4 fast growing and 4 slow growing.

Components of the preparations: transparent liquid cultures of biologically active microorganisms (BAM) with a cultural sediment in glass test tubes (diameter 20 mm, length 200 mm), marked with an inscription and sealed with rubber stoppers. The volume of culture liquid - 10 ml. The total number of test tubes - 10 pcs. The test tubes were divided into two groups: 1) 4 test tubes for the preparation of Agrobiovit P1 and 2) 6 test tubes for preparation - Agrobiovit P2. The concentration of the working solution is 10 ml of each BAM suspension per 10 liters of sterile water.

Labeling tubes with liquid cultures. For the preparation of Agrobiovit P1: Az.ch., V.r., Ps.Pl, Tr.ho. For the preparation of Agrobiovit P2: Az.ch., V.r., Ps.Pl, Tr.ho., R. o., B.s.

Scheme for preparing preparations: BAM cultures were thoroughly mixed before obtaining preparations, obtaining a uniform opalescent cultural suspension. To obtain working solutions of the components, 10 ml of each BAM culture was brought to 10 l with sterile water. To obtain preparations, working solutions of the components were mixed in equal proportions.

The studies were performed according to the generally accepted methodology: Minaeva O.M., Akimova E.E., Zyubanova T.I., Tereshchenko N.N. Biopreparations for plant protection: assessment of quality and efficiency: textbook. allowance. - Tomsk: Publishing House of Tomsk State University, 2018. - 130 p.

Setting up an experiment.

The experimental scheme included 5 options: control; chemical standard - drug Delan, VG (700 g/kg of dithianon), which is a contact fungicide; biological standard - the drug Fitosporin-M, the active component of which is the strain 26 D of the bacterium Bacillus subtilis, the titer is not less than 1 billion living cells and spores / ml; tested preparations - innovative multicomponent biological preparations Agrobiovit P1 and Agrobiovit P2. The repetition in the experiment is fourfold. The total number of inoculations was 20. The analysis time was 40 days, not including obtaining a pure culture of the pathogen.

Working solutions of preparations were applied to the surface of nutrient agar in Petri dishes and evenly distributed with a sterile spatula. After that, Petri dishes were placed in a thermostat at a temperature of 26°C for the growth of the active components of biological preparations and the absorption of the active substance into agar-agar in a chemical preparation. Sterile distilled water was applied to the control Petri dishes. Three days later, the surface of the nutrient agar was covered with filter paper soaked in sterile water, and the agar plates were turned upside down. Agar blocks 5 mm in size cut from various isolates of the pathogen V. inaequalis were laid out on the control and experimental media thus obtained. 4 agar blocks were placed in each Petri dish (opposite each other). A total of 8 different isolates were analyzed.

Obtaining a pure culture of Venturia inaequalis was carried out on potato glucose agar (CGA) for 30 days at a temperature of 20°C in the dark.

The antifungal efficacy of all drugs was evaluated in two ways:

1) according to the intensity of the radial growth of the pathogen culture on the 30th day of the experiment, measured in millimeters and expressed in biological efficiency relative to the control variant;

2) according to the degree of development of aerial mycelium on the seed block of the pathogen, assessed on a five-point scale. After evaluating the radial growth, the size of the seed block was subtracted from the obtained values.

Evaluation scale for the development of mycelium on the sowing block, points: 0-0%; 1 - 10%; 2-25%; 3-50%; 4 - 100%. The density of aerial mycelium development was estimated according to the degree of coverage of the sowing block with fungal hyphae.

Statistical processing and validation of the received data. For each tested pathogen isolate, the biological effectiveness of the drug (BE) was calculated using the formula:

BE \u003d (A-B) / A * 100,%,

where: BE - biological efficiency;

A - the number of harmful organisms before processing;

B - the number of harmful organisms after treatment.

To confirm the presence of the effect of Agrobiovit P1 and Agrobiovit P2 preparations on the growth of pure cultures of scab isolates in laboratory conditions, an analysis of variance was carried out on the data obtained, and in the presence of the effect, NSR ₀₅ was calculated - the smallest significant difference at a significance level of 0.05.

Results.

All variants with the use of drugs showed an inhibitory effect on the radial growth of the pathogen on all or several isolates of the apple scab pathogen, but with different biological efficiency.

On the chemical standard, a significant inhibitory effect against Venturia inaequalis was shown only for 5 out of 8 isolates. These were all fast growing isolates and one slow growing isolate (Table 1). The biological effectiveness of Delan at a sufficient level (83%) was found only for isolate 11_1; four isolates were found to have insufficient biological effectiveness. For two slow-growing isolates 7_5 and 8_1, a stimulating effect was detected, and for isolate 8_2, no effect was found.

Fitosporin-M produced a significant inhibitory effect for all fast-growing and two slow-growing strains of the pathogen (7_4 and 7_5, Table 2). At the same time, only for 3 isolates the effectiveness of the biological preparation was 100% (10_2_1, 11_1 and 22_2) and for another 92% (11_3, Table 2).

The test preparations Agrobiovit P1 and P2 significantly inhibited the radial growth of all test isolates of Venturia inaequalis with high biological efficiency: 100% (Table 3). In Petri dishes, the growth of both substrate and aerial mycelium of the isolates was completely absent. The effectiveness of both drugs was equally high.

Table 4 presents the results of evaluating the antifungal activity of preparations for the development of aerial mycelium on the sowing block. The highest degree of aerial mycelium development was characteristic of the control variant and the chemical standard (4 points, Table 4).

In the Fitosporin variant, there were several variants from oppressed to medium development of aerial mycelium, from 1 to 3 points. The highest antifungal activity was noted for the experimental preparations Agrobiovit P1 and P2, on which the development of aerial mycelium on the seed block in the pathogen did not occur and was equal to 0 points.

Thus, the results of two approaches to assessing antifungal activity, according to the rate of radial growth and the degree of development of aerial mycelium on the seed block, coincided.

The study showed the presence of fungicidal properties in the preparations Agrobiovit P1 and Agrobiovit P2 in relation to all eight tested isolates of the apple scab pathogen with 100% biological efficiency and exceeding the effects of chemical and biological standards, Delan, VG and Fitosporin-M, Zh, respectively. Differences in the degree of fungicidal activity between the tested preparations Agrobiovit P1 and Agrobiovit P2 were not revealed.

Example 2. Conducting experimental studies to study the effectiveness of the use of microbiological fungicides Agrobiovit P1 and Agrobiovit P2 to protect apple plants from scab Venturia inaequalis (Cke.) Wint.

Period of the experiment: June-July 2021

Type of experience: small-plot field.

The scheme of experience is presented in table 5.

As a standard, highly effective preparations against apple scab were used: the microbiological preparation Fitosporin-M, Zh (titer of at least 1 billion living cells and spores / ml), the active principle of Bacillus subtilis, strain 26 D, and the fungicide of chemical genesis Delan, VG (700 g /kg dithianon).

Plot size and placement: 4 trees, in blocks.

Number of repetitions: 4.

Variety: Renet Simirenko - late ripening, highly susceptible to scab.

Methodology: The following guidelines were used for testing: Guidelines for registration tests of fungicides in agriculture. - St. Petersburg, 2009. - 266 p., Armor B.A. "Methodology of field experience". - M., 2014

Results.

On the experimental plot, the scab had an epiphytotic character of development during the main growing season. A high rate of scab infection was noted in the second or third decade of June.

leaf protection

Before the application of biofungicides, the spread and intensity of scab development in the treated variants were the same: on June 17, 1.50% of the leaves (disease spread) were affected with a disease development intensity of 0.65%, on control trees - 13.00% and 10.96%, respectively. The biological effectiveness of protection was 94.1%.

After the first application of biological preparations on the 5th day (June 23), the effectiveness of protection in all variants, in comparison with the efficiency on June 17, decreased, but slightly: by 2.2-7.3% (Table 6). In the control during this period, a high rate of infection was noted: for 5 days, the number of affected leaves increased by 1.4 times. Between the options, the difference in efficiency was obtained insignificant: from 3.9 to 5.1%. Thus, within 5 days after treatment, the scab was contained quite effectively in all cases.

The action of biological preparations proceeded in quite favorable conditions: the maximum air temperature was within 28.4-32.3°C, the average temperature was within 21.5-26.2°C, the average relative air humidity was within 64-78%.

On the 10th day (August 28), a significant decrease in the effectiveness of protection was noted in all variants. The spread and intensity of the development of the disease for the period June 23-28 in all variants increased: in the Agrobiovit P1 variant - by 1.9 and 2.2 times, in the Agrobiovit P2 variant - by 2.3 and 3.7 times, in the Delan variant - in 1.7 and 3.0 times, in the Fitosporin-M variant - 2.1 and 3.7 times. From June 23 to June 28, an abnormal amount of precipitation fell: 28.8 mm, which, according to long-term average data, was 76% of the ten-day norm, and, of course, affected the effectiveness of fungicides.

At the same time, in the control variant from June 23 to June 28, the increase in the infectious background was insignificant. At a low rate of infection by the 10th day after application, the effectiveness of fungicides in all variants was insufficient. The best scab control was provided in the Agrobiovit P1 variant, which corresponded to the effectiveness of Fitosporin-M. The Agrobiovit P2 variant was superior in efficiency to the second standard - Delan, VG. The difference in biological efficiency between the variants of application Agrobiovit P1 and Agrobiovit P2 was 13.0%.

After the second application of biological preparations on the 5th day (03.07), in all variants, the effect of blocking a new infection was noted - the absence of infection of young leaves on the shoot. The spread and intensity of the development of the disease for the period 28.06-03.07 in the variants decreased, respectively: in the Agrobiovit P1 variant - by 1.5 and 1.4 times, in the Agrobiovit P2 variant - by 1.6 and 1.4 times, in the Delan variant - by 1.7 and 1.8 times, in the Fitosporin-M variant - by 1.5 and 1.4 times. In the control variant, from June 23 to June 28, the increase in the infectious background was insignificant. This is due to insufficiently favorable weather conditions for the development of scab. Abnormal heat was noted: from 01.09 to 03.09 the maximum air temperature reached 33.8-34.6°С; there were no precipitations, the relative humidity of the air from 01.09 to 03.09 dropped to 31-40%. Thus, with a stable infectious background, the second sequential treatment with biofungicides made it possible to restrain the development of scab on the leaves.

On the 10th day (08.07) after the second treatment, only in the variant of application of Agrobiovit P2, there was no containment of the development of the infection: the spread and intensity indicators in this variant increased by 1.4 and 1.2 times, respectively. In the other three variants, these indicators practically did not change, that is, the infection was blocked. A difference was noted between the Agrobiovit P1 and Agrobiovit P2 variants: the spread of the disease on the leaves in the Agrobiovit P1 variant was 1.5 times lower, the intensity of development was 1, 8 times; efficiency is higher by 10.7%. A comparative analysis of the application of the fungicide Agrobiovit P1 with both standards showed that close values of the P and R indicators were obtained, and, consequently, close protection efficiency.

In the control variant, from 03.07 to 08.07, the indicator of the intensity of the development of the disease increased: 1.3 times. The increase in the disease was associated with a large amount of precipitation in a short period of time: from 04.07 to 07.07 16 mm; the average relative humidity these days was 78-93%.

The results obtained after the second treatment confirm the effectiveness of Agrobiovit P1 Agrobiovit P2 against apple scab under conditions of a stable infectious background.

Thus, as a result of the field experiment, the effectiveness of protection against leaf scab was established for the options for the use of biofungicides Agrobiovit P1 and Agrobiovit P2.

Fruit protection.

Before the application of biofungicides, the spread and intensity of scab development on fruits in the treated variants were the same: on June 17, 4.25% of the fruits were affected (disease spread) with a disease development intensity of 1.38%, on control trees - 29.91% and 14.87, respectively. %, the protection efficiency was thus 90.7%.

After the first application of biological preparations on the 5th day (June 23), the effectiveness of protection in all variants, in comparison with the effectiveness on June 17, decreased: in the Agrobiovit P1 variant - by 3.9%, in the Agrobiovit P2 variant - by 8.2%, in in the Delan variant - by 6.7%, in the Fitosporin-M variant - by 11.0% (Table 7). From an economic point of view, this is significant. In the control during this period, a high rate of infection was noted: for 5 days, the number of affected fetuses increased by 1.5 times, the intensity of the development of the disease - by 1.9 times. Thus, at a high rate of infection and against a high infectious background, none of the fungicides, including those of chemical origin, showed blocking of the infection. A difference was noted between the Agrobiovit P1 and Agrobiovit P2 variants: the spread of the disease on fruits in the Agrobiovit P1 variant was 1.5 times lower, the intensity of development was 1.3 times lower.

On the 10th day (August 28), a significant decrease in the effectiveness of protection was also noted in all variants. The spread and intensity of the development of the disease for the period June 23-28 in all variants increased, while almost the same: in the Agrobiovit P1 variant - by 3.3 and 3.2 times, in the Agrobiovit P2 variant - by 3.4 and 3.3 times, in the Delan variant - by 3.6 and 3.3 times, in the Fitosporin-M variant - by 3.3 and 3.0 times. In the control variant from 23.06 to 28.06, the increase in the infectious background was also significant: the number of affected fetuses increased by 1.8 times, the intensity of their damage - by 1.4 times. Thus, at a high rate of infection by the 10th day after application, the effectiveness of protection against scab in all variants was insufficient. The best scab control was provided in the Agrobiovit P1 variant, which exceeded the effectiveness of both standards. The efficiency of Agrobiovit P2 exceeded the efficiency of Fitosporin-M and corresponded to the efficiency of Delan, VG. The difference in biological efficiency between the variants of Agrobiovit P1 and Agrobiovit P2 was 11.7%.

After the second application of biological preparations on the 5th day (03.07), the same trend was noted: the absence of complete containment of the disease in all cases. From 28.06 to 03.07 in the variants of application of Delan, VG and Fitosporin-M, the intensity of development increased by 1.3 times, in the variants of Agrobiovit P1 and Agrobiovit P2 - by 1.7 times. In the control variant, from June 23 to June 28, the increase in the infectious background was insignificant.

On the 10th day, the increase in the number of affected fetuses was the largest - 1.4 times - in the Agrobiovit P2 variant, the increase in development intensity - also 1.4 times - in the variants of Delan, VG and Fitosporin-M. Thus, the best performance was obtained in the Agrobiovit P1 variant; the most noticeable differences are in the indicator of biological efficiency, which is 1.2-1.5 times higher in comparison with other options for Agrobiovit P1.

Phytotoxicity to culture. The presence of signs of acute and chronic phytotoxicity to the experimental fungicides Arobiovit P1 and Agrobiovit P2 on the leaves, shoots and fruits of the Renet Simirenko apple tree variety was not observed.

Conclusions about test results

1. Experimental biofungicides Agrobiovit P1 and Agrobiovit P2 were tested on apple varieties highly susceptible to scab under conditions of disease epiphytotics. Biofungicides were used in two successive treatments: in the phases of apple tree development "fruit diameter up to 40 mm, erect fruit" and "about 50% of the cultivar-typical fruit size achieved".

2. In the field, the effectiveness of Agrobiovit P1 and Agrobiovit P2 against apple scab was established. The level of biological effectiveness of Agrobiovit P1 in protecting leaves was not inferior to the level of effectiveness of both standards - the chemical fungicide Delan, VG and the microbiological preparation Fitosporin-M, Zh, and when protecting fruits - exceeded their level. Agrobiovit P2, when protecting leaves, was slightly inferior to both standards, while protecting fruits, it had a similar level of efficiency with them.

3. The period of protective action of Agrobiovit P1 Agrobiovit P2 from scab conidial infection was 5 days, which is close to the usual period of protection from the disease for biofungicides (7 days).

4. The effectiveness of the mixtures increased after the second application, therefore, it is advisable to apply the mixture in a block of two treatments.

5. Experimental biofungicides Agrobiovit P1 and Agrobiovit P2 have the prospect of being used as a fungicide

Claims (4)

1. Composition for protecting plants from apple and/or pear scab containing Trichoderma viride, Azotobacter chroococcum, Bacillus pumilus, Pseudomonas fluorescens. 2. Composition according to claim 1, additionally containing Bacillus subtilis and Pythium oligandrum. 3. Composition according to claim 1 or 2, wherein the plant is an apple tree. 4. Composition according to claim 1 or 2, wherein the plant is a pear.