Method to stimulate growth and protect small-fruit crops against diseases caused by fungic pathogens

FIELD: agriculture.

SUBSTANCE: method to stimulate growth and to protect small-fruit crops against diseases caused fungic pathogens includes treatment of plants with a biopreparation in a liquid form by soaking roots of seedlings prior to planting, or their sprinkling after planting, or sprinkling of plants in the period of vegetation and fruiting, or sprinkling of soil around plants with its further soaking with water. The biopreparation is a mixture of strains of bacteria B. subtilis VKPM B-10641, B. amyloliquefaciens VLPM B-10642, B. licheniformis VKPM B-10561 and B. licheniformis VKPM B-10562 or a mixture of strains of bacteria B. subtilis VKPM B-10641, B. amyloliquefaciens VKPM B-10642 and B. licheniformis VKPM B-10562 with a titre of each strain of not less than 105 CFU/ml in the form of a water suspension.

EFFECT: invention provides for higher protection of small-fruit crops against diseases caused by fungic pathogens, due to use of a mixture of strains of different types of bacteria of Bacillus type, having higher antagonistic activity, and also provision of the possibility of stimulation of growth of small-fruit crops.

5 cl, 10 tbl, 8 ex

 

The invention relates to methods of promoting growth and protecting fruit crops from diseases caused by fungal pathogens, and can be used in agricultural Microbiology, agricultural biotechnology, plant protection.

There is a method of cultivation and plant protection (Patent RF №2154935, IPC A01G 1/00, publ. 27.08.2000 year), including soil preparation, planting or sowing of the plants, periodic cultivation, and is characterized by the fact that they are carrying out pre-processing of seeds and tubers with a mixture of activators, Immunostimulants, antagonists and biofertilizers [Overcrop, agate-25 or Rhizoplane, Ampelomyces, Gerbamin, Disprin or Nematol, Complex biological fertilizer (BMF), the Symbiont-Estate] in half the dose recommended for this product (mixture 1), the same mixture in combination with biological fertilizers (Vermicompost, Root mixture at recommended doses) are inserted in holes before planting, the soil together with the biological make waste Trichogramma production or waste viral biologics (dead butterflies) with sapropel or rotted chicken manure, or with powder from naturally fallen leaves or needles (mix 2), further a mixture of 1 conduct periodic ground-based spraying or irrigation of vegetative plants (every two weeks) with pre-razbrasivayutsya and the introduction into the soil of the Root mixture (same rules), soil inhabit a range of microorganisms, microbial antagonists contained in the biologics [Alien-In, Alien, Pacific, Gliocladium, Mycostop, Narcissus, Nematology or Nematology (activated supernatant is away from the production of a biological product, Disprin), Pentothal, Isoplan-f, various forms of Trichodermin, Trichothecin, Vitaflavan, Vitaflavan-L] (mixture 4), at half the dose recommended for this drug, which contribute in the hole before planting, then periodically (1 every two weeks) spraying or watering plants; enter as elements of the fauna of predators and entomophages (Aphidius, Amblyseius, ladybirds, Verbliudka, Midge, Lacewing, Lysiphlebus, Sifredi, Phytoseiulus) - complex 1 - by periodic release of half doses recommended for this drug and microorganisms, which is the current top of biologics (Overcrop, Verticalin, Cephalosporins, Endomorphin, Bicol, Methoxylation, Bowery, Vertimax, Mihaylin-T, Torild, Entertain) (mix 5), at half the dose recommended for this drug by periodic treatment - spraying or watering; before spraying Diprima and/or Namatala use sodium alginate to prevent drying and death of nematodes, and the use of entomophages, at the beginning of the growing season is about cultivation of agricultural crops, additionally implanted into the aisle staple crops Compositae and/or umbrella plants, which are an additional source of food for adult entomophages.

The disadvantage of the above is similar complexity and costs playback method of plant protection.

There is a method of biological protection of plants on the example of potatoes, including the processing of tubers and potato plants during the growing season a biological product in a mixture with adhesive, characterized in that the processing of potato tubers as a biological product used taking into consideration of the calculation of 1000 ml per tonne of tubers, and to treat 1 ha of planting during the growing season using a mixture of biological products taking into consideration the amount of 0.5 l and bankol in the amount of 0.2 l, dissolved in 300-350 liters of mineral water "Samanco", as the adhesive used corn starch in the amount of 1-1,5% of the volume of the mixture (patent of the Russian Federation No. 2343667, IPC AS 1/00, A01N 63/00, publ. 20.01.2009,).

However, in this method there is no data about the opportunity to stimulate growth and protect fruit crops from diseases.

There is a method of growing plants on depleted in nitrogen and micronutrients to the soil, including the processing of seeds and/or vegetative plants biological preparation containing a suspension of bacteria strain Pseudomonas, characterized in that as beapr the parathas use a mixture of suspensions of strains of Pseudomonas species 7G, GC and 17-2, taken in the ratio 1:2:1 with a concentration of 2.5 to 5.0×1010cells/ml, while the treatment of the seeds produced from the calculation of 30-50 ml of the suspension per 1 ton of seeds, and processing plants in active growth phase, the rate of 30-75 ml/ha (RF patent No. 2403748, IPC A01N 63/00, publ. 20.11.2010,).

However, in this method there is no data about the opportunity to stimulate growth and protect fruit crops from diseases.

There is a method to stimulate growth and protect plants from diseases, which includes seed treatment composition of the bacterial cells of the microorganism of the genus Pseudomonas in the form of a complex of strains of Pseudomonas Species 7G, Pseudomonas Species 7G 2K, Pseudomonas Species 17-2 and physiologically active components, characterized in that the physiologically active components use humic fertilizer "Darina", the ratio of bacterial strains is 0,7:1,5:0,7-1,5:0,7:1.5, the concentration of bacterial cells 2-5·1010cells/ml, the ratio of the complex strains and "Darina" is 1:1-1:1,8, composition prepared before processing seed breeding in culture liquid of a three-day Chlorella Vulgaris, and for processing 1 ton of seeds using 2,3-2,7 ml of the complex strains of bacteria and the rate of flow of the working solution is 10-12 litres per tonne of grain and leguminous crops and 70-80 litres per tonne of potato tubers (RF patent No. 2291620, IPC A01G 5/00, publ. 20.01.2007,).

However, in this method also the data are present about the opportunity to stimulate growth and protect fruit crops from diseases.

The closest analogue (prototype) is a method of protecting fruit and berry crops from diseases, including the processing of the drug in liquid form, containing a mixture from the culture fluid of the bacterial strain Bacillus subtilis 24 D (ARRIAM 129) at a concentration of 5·109cells/ml, humates and trace elements, described in the patent of the Russian Federation (RF patent No. 2216173, IPC A01N 63/00, publ. 20.11.2003 year). The method is used on berry bushes, fruit trees. Taken 50 ml and diluted with two liters of water. This solution is poured 2-3 tree/shrub or 100 m2planting strawberries.

However, this method has the lack of efficiency of protection of fruit crops due to the use of the drug based on a single strain of bacteria Bacillus subtilis 24D (ARRIAM 129), and there is no evidence of stimulation of plant growth using the prototype method.

The technical result is to increase the protection of fruit crops from diseases caused by fungal pathogens through the use of a mixture of strains of different species of bacteria-probiotics with high antagonistic activity and more diverse forms of processing plants, as well as providing the possibility of stimulating the growth of berries.

This technical result is achieved in that in the method of promoting the growth and protection of fruit crops from bol is SNA, caused by fungal pathogens, including the processing of berry crops biological product in liquid form, according to the invention the plants are treated by soaking the roots of the seedlings before planting, or spray them after planting, or spray plants during the growing season and fruiting, or spray the soil around the plants with subsequent moistening with water, and as a biological product, a mixture of bacterial strains .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642, .licheniformis VKPM B-10561 and .licheniformis VKPM B-10562, or a mixture of strains of bacteria .subtilis VKPM B-10641; B. amyloliquefaciens VKPM B-10642 and .licheniformis PMBC In-10562 with a title for each strain of not less than 105CFU/ml in aqueous suspension. By stimulating the growth and protection of strawberries from fungal diseases it landing in the soil is carried out after soaking the roots of the seedlings in the aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: strain .licheniformis VKPM B-10562; strain .subtilis VKPM B-10641; strain .amyloliquefaciens VKPM B-10642 with a title for each strain, equal to 105CFU/ml, and fruiting strawberry plants are sprayed with an aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: .licheniformis VKPM B-10562; .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642 with a title for each strain, equal to 106CFU/ml

By stimulating the growth of buckthorn seedlings sprayed the soil is in between the plants with an aqueous suspension of the drug, containing a mixture in equal proportions of bacterial strains: .licheniformis VKPM B-10562; .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642 with a title for each strain, equal to 106CFU/ml, followed by moistening the soil to a depth of 10-15 cm irrigation water.

When the protection planting raspberries or blackcurrants from diseases landing sprayed with an aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains .licheniformis VKPM B-10561; .licheniformis VKPM B-10562; .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642 with a title for each strain, equal to 106CFU/ml, with a flow rate of the aqueous suspension of 500 l per 1 hectare

When stimulated seedling growth of black currant sprayed the soil between the plants with an aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: .licheniformis VKPM B-10561, .licheniformis VKPM B-10562; .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642 with a title for each strain, equal to 106CFU/ml with subsequent moistening soil between the plants to a depth of 10-15 cm irrigation water.

Used in the inventive method, the bacterial strains belong to the applicant and deposited in Russian national Collection of Industrial Microorganisms FSUE gosniigenetika:

Bacillus subtilis for the restoration of microflora of the soil and the gastrointestinal tract of animals with bactericidal, fungicidal and virucidal activity and Deposit the p in the all-Russian Collection of Industrial Microorganisms FSUE gosniigenetika under registration number VKPM B-10641 (certificate of Deposit attached);

Bacillus amyloliquefaciens VKPM B-10642 for the restoration of microflora of the soil and the gastrointestinal tract of animals, which have bactericidal and fungicidal activity and deposited in Russian national Collection of Industrial Microorganisms FSUE gosniigenetika under registration number VKPM B-10642 (certificate of Deposit attached);

- strains of Bacillus licheniformis for the restoration of microflora of the soil and the gastrointestinal tract of animals, which have bactericidal and fungicidal activity and deposited on 18.03.2010, in the all-Russian Collection of Industrial Microorganisms (VKPM) FSUE gosniigenetika, respectively, under the number VKPM B-10561, 10562.

The following is a description of the proposed method.

The way to stimulate growth and protect fruit crops from fungal diseases is as follows. The method includes processing and berry crops biological product in liquid form by soaking the roots of the seedlings before planting, or spray them after planting, or spray plants during the growing season and fruiting, or spray the soil around the plants with subsequent moistening with water.

As a biological product use the following options:

Preparation No. 1. A mixture in equal proportions of bacterial strains .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642 and .licheniformis PMBC is -10562 with a title for each strain of not less than 10 6CFU/ml in aqueous suspension.

Preparation No. 2. A mixture in equal proportions of bacterial strains .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642, and .licheniformis VKPM B-10562 with a title for each strain of not less than 106CFU/ml in aqueous suspension.

Preparation No. 3. A mixture in equal proportions of bacterial strains .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642, B. licheniformis VKPM B-10561 and .licheniformis VKPM B-10562 with a title for each strain of not less than 106CFU/ml in aqueous suspension. By stimulating the growth and protection of strawberries from disease its planting in the ground is carried out after soaking the roots of the seedlings in water suspensions of drug No. 1 based 9-10 l aqueous suspension at 3000 roots of strawberries. A soak time of one party the roots of the strawberry is 2 hours.

To protect fruit-bearing strawberry plants, for example, against Botrytis her sprayed with an aqueous suspension of the drug No. 2. The flow rate of the working fluid at the rate of 500 l/ha 50 ml/m

By stimulating the growth of buckthorn seedlings sprayed the soil between the plants with an aqueous suspension of the drug is No. 1 at the rate of 40 liters of water suspensions of the drug on 4000 roots with subsequent moistening the soil to a depth of 10-15 cm irrigation water.

When the protection planting raspberries or blackcurrants from diseases landing sprayed with an aqueous suspension of the drug No. 3 with the consumption of water suspension of 500 l per 1 hectare

Stimulation of restressed black currant sprayed the soil between the plants with an aqueous suspension of the drug No. 3 at the rate of 700 roots of seedlings per 10 l of water suspension of the drug with subsequent moistening soil between the plants to a depth of 10-15 cm irrigation water.

Below are the experimental data in examples 1-8, confirming the efficiency of the proposed method.

Example 1

They studied the effectiveness of bacterial strains of the drug No. 1 used in the present method, when the cultivation of the strawberry varieties Junius smayds influence on growth, development and the phytosanitary condition of the plantation culture. The dates of planting strawberries from the 3rd decade of may until mid June. The concentration of the applied strains in the advanced options is 105CFU/ml the Number of plants 3000 pieces per option 1 (one strain). The method of application of strain - soaking the root system of the seedlings strawberries in the working fluid containing the strain at a concentration of 105CFU/ml, the Flow rate of the working fluid on option 1 to 9 liters. Exposure time is 2 hours.

We studied the effect of strains on the survival, growth and development of young plants and defeat them in white spotting. For bookmarks of experience used plants grown in the mother liquor of the SHA "Gardens of Siberia". The excavated seedlings root system was immersed in the solutions of the studied strains for 2 hours prior to boarding to a permanent place on the industrial quarter of the economy.

At the end of the vegetation period, the degree of the damage to plants white blotch (caused by Ramularia tulasnei Sacc, Hyphomycetales, Deuteromycota) when processing strains .amyloliquefaciens VKPM B-10642, .licheniformis VKPM B-10562, .subtilis ITCM IN-10641 was decreased 2.0-2.4 times.

On young leaves, which grew up after landing on the production quarter, the degree of the damage was very low (0.1 to 1.9 per cent, there was observed a single spot. The prevalence of symptoms was significantly decreased in the variants with the use of strains of options using strains of B. subtilis ITCM IN-10641 and .licheniformis VKPM B-10562, respectively 7.2 and 9 times. This indicator biological efficacy of the treatment were, respectively, 86 and 89%.

Thus, treatment of bacterial strains .amyloliquefaciens VKPM B-10642, B. licheniformis VKPM B-10562, .subtilis ITCM IN-10641 drug No. 1 had antidepressive impact and improve the sustainability of strawberry plants to the white spot.

Shows a significant treatment effect of bacterial strains on the performance of plant biomass. The total biomass of 1 plant was significantly increased in 1.7 times (table 1) in the variant with application of strain Century amyloliquefaciens VKPM B-10642 (23,2 g) relative to control (13,4 g). After removal from the plant of old leaves formed before the experimental treatment, differences emerged even more clearly. In the control biomass per plant was 7.0 g; in the variant strain Century amyloliquefaciens VKPM B-10642 - 12.5 g (an increase of 1.8 times); variations is the strain .subtilis ITCM IN-10641 - 12.2 g (1.7 times); in the variant strain .licheniformis VKPM B-10562 - 11,7 g (1.7 times).

In the variants with strains .subtilis ITCM IN-10641 and .licheniformis VKPM B-10562 also found significant (P<0.05) increase in the biomass of the root system (rhizomes and roots) in 1,4-1,5 times.

The length of the aerial parts was significantly increased in the variant strain .amyloliquefaciens VKPM B-10642 (1.4 - 7.0 cm) and with strain .licheniformis VKPM B-10562 (by a factor of 1.2 - 3.1 cm).

The length of the root system was significantly increased in variation with strain .amyloliquefaciens VKPM B-10642 (14%) and in the variant strain .subtilis ITCM IN-10641 (21%).

Table 2
The influence of bacterial strains of drug # 1 on the biomass and the length of the strawberry plants in the manufacturing experience (SHA "Gardens of Siberia", September 14, 2010)
Biomass, gLength, cm
Optiontotalwithout old leavesthe root systemaerial partsroots
Control (without about the creation) the 13.47,03,219,115,1
.amyloliquefaciens VKPM B-1064223,2*12,5*4,226,1*17,2*
In subtilis ITCM IN-1064118,612,2*4,9*22,818,2*
.licheniformis VKPM B-1056219,011,7*4,5*23,2*16,4
The drug is No. 1 on the basis of a mixture of strains20,312,1*4,5*24,0*17,3*
NDS056,53,71,1a 3.92,0

Thus, the obtained data show the presence of a catalyst of action in all 3 tested strains of drug # 1 on various pairs of the points of the growth and development of strawberry plants, and the defeat of the white spot.

Example 2

The positive influence of strains .amyloliquefaciens VKPM B-10642, .subtilis ITCM IN-10641 drug No. 1 when landing on rooting woody cuttings of sea buckthorn. The dates of planting cuttings buckthorn experience : 2nd decade of may. The concentration of the applied strains in the advanced options is 105CFU/ml Number of cultivated plants - 4000 pieces per option 1. The method of application of bioagents - spraying the soil surface with the working fluid containing bioagent, followed by moistening the soil to a depth of 10-15 cm by irrigation water. The hydraulic fluid option 1 - 40 litres.

Action test bacterial strains to become rooted cuttings of sea buckthorn were performed on plant varieties are Plentiful.

The cuttings buckthorn was not detected malicious background of the development of diseases or pests during the growing season.

Revealed that the length of annual growth of aboveground parts (along the length of the main stem) in the control was 28,4 cm (table 3; figure 1). Under the influence of the processing strain .amyloliquefaciens VKPM B-10642 was statistically significant (P<0.05) elongation growth of 3.8 cm (20%), under the influence of the strain B. subtilis ITCM IN-10641 - 4.3 cm (22%).

The length of the root system in the control was 26.2 see When processed by a strain .amyloliquefaciens VKPM B-10642 root system reliably UD is inalas 6.9 cm (26%), when processed by a strain of B. subtilis ITCM IN-10641 - 8.9 cm (34%).

Table 3
The treatment effect of bacterial strains of drug # 1 on the parameters of the growth and development of woody cuttings buckthorn (sort Abundant kennel SHA "Gardens of Siberia", accounting, September 25)
OptionLength of annual growth, cmThe length of the root system, cmThe average biomass of 1 plant, g% of control
Control (without treatment)28,426,212,2-
.amyloliquefaciens VKPM B-1064234,2*33,1*16,3*33,6
.subtilis ITCM IN-1064134,7*35,1*16,3*33,2
.licheniformis VKPM B-1056228,030,212,633,
The drug is No. 1 on the basis of a mixture of strains32,3*32,8*15,133,3
NDS055,04,44,1

Biomass per plant in the control was 12.2, options processing strains .amyloliquefaciens VKPM B-10642 and .subtilis ITCM IN-10641 biomass 1 plants reached 16.3 g (an increase of 33-34%).

Thus, strains .amyloliquefaciens VKPM B-10642 and .subtilis ITCM IN-10641 drug No. 1 provided for rooting woody cuttings buckthorn expressed stimulating effect.

Example 3

Model experience on fructiferous stocky strawberries on the square 20 m2the effect on grey mould on fruits. Option for each strain of drug No. 2 includes 20 fruiting strawberry plants located in 2 adjacent rows in a square 1 m2and the control without treatment. The biological agents are applied by spraying at the beginning of fruit ripening 3rd decade of June (for 3-5 days to 1st harvest). The concentration of the used strains of drug No. 2 in the advanced options is 106CFU/ml, the Flow rate of the working fluid at the rate of 500 l/ha 50 ml/m2. Estimated individual plants in popcorn the STI 20 copies of each version.

In a model experiment to test the validity of the bacterial strains of the drug No. 2 on the defeat of the fruit of the strawberry gray mold (caused by Botrytis cinerea Pers., Hyphomycetales, Deuteromycota) processing plants held on June 26 in the phase of ripening beginning of ripening. Experienced plant varieties Junius smayds (20 plants in each case) were sprayed once, the drug was applied to the entire above-ground part and on the soil surface in rows. The first ripe fruit was formed on July 2-3. Records destruction carried out in 2 time, before mass charges berries - July 10 and July 15. Valued at each period of 250-300 fruit in each case.

In 1st period (July 10) under the influence of the strain B. subtilis ITCM IN-10641 prevalence berries grey mould significantly (P<0,05) decreased 1.7 times (EB=41%), under the influence of strain Century licheniformis VKPM B-10562 - 2.5 times (EB=61%). In the 2nd period (15 July) the effect of the strains showed still more clearly. All 3 of the test strain significantly reduced the lesion grey mould from 2.2 to 3 times, with EB=54-61%.

Thus, all 3 tested strains of drug No. 2 showed similar efficacy against Botrytis fruit of the strawberry: strain .amyloliquefaciens VKPM B-10642 had a pronounced effect on the growth of aboveground parts and an increase in the biomass of wild strawberry and sea buckthorn, on the growth of the roots of sea-buckthorn, and this strain was reduced defeat Mr. gray is pour strawberries; strain .subtilis ITCM IN-10641 increased biomass and length of the roots of strawberry and berries, and increased the length of the annual increment of above-ground system at sea buckthorn, reduced the incidence of shot hole leaf and affection of strawberry grey mould; strain .licheniformis VKPM B-10562 caused an increase in the biomass of the root system of wild strawberry, effectively influenced against fungal shot hole Skytrain system of strawberries, as well as against gray rot of strawberries.

Example 4

To protect fruit crops during the growing season from fungal diseases conducted twice processing raspberry varieties Zorenka Altai with an aqueous suspension of the drug No. 3 (.licheniformis VKPM B-10561; .licheniformis VKPM B-10562; B. subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642) against purple blotch. As control was used plants treated with water.

The results of drug and No. 3 on the development of the purple spot in the field experiment are presented in table 4.

Under the influence of biologics prevalence and development of disease decreased 1.5-1.7 and 1.6-1.8 times compared with the control. Deep defeat raspberry canes this year was not marked. The development of the disease in the control did not exceed the economic threshold (ET) (ET - 25%). The yield increase of 0.1 t/ha

Thus, the tests showed that the strains .licheniformis - PMBC-10561 and VKPM B-10562 and strains .subtilis In the PM-10641, .amyloliquefaciens VKPM B-10642 drug No. 3 actively inhibit the development and spread of purple spot and can be agents of biological control of the pathogen purple gummy raspberries.

Table 4
The influence of strains of B. subtilis, B. licheniformis and Bacillus amyloliquefaciens drug No. 3 defeat raspberry purple spot
OptionPrevalence, %Development %Yield, t/ha
Control75.018,71,4
Strain .licheniformis-PMBC-1056242,510,61,5
Strain .licheniformis-PMBC-1056145,011,31,5
Strain B.subtilis VKPM B-1064142,511,91,5
Strain Century amyloliquefaciens VKPM B-1064247,511.9 1,5
The drug is No. 3 on the basis of a mixture of strains44,411,41,5
Topaz 0,1%4011,31,5
NDS053,0

Example 5

Processing raspberry directed against the underlying disease culture-purple spot at the first symptoms of the disease. Processing single - when the first symptoms of the disease. The concentration of the used strains of drug No. 3 in the advanced options is 106CFU/ml. the Method of application of bioagents - ground mechanized spraying (MT3-80+OECS-2200). The flow rate of the working fluid at the rate of 500 liters per 1 ha - 125 l option.

In the manufacturing experience to test the validity of bacterial strains to failure raspberry fungal diseases treatments were carried out on 15 July 2010, the Drug was applied to the plants tractor sprayer OECS-2200.

At the time of processing plants was observed individual symptoms of Septoria leaf blotch (caused by Septoria rubi (West.) Sacc, Spaeropsidales, Deuteromycota) on the leaves. By the date of 1st registration, held on 28 July, the degree of destruction is aphorism in the control reached 10%, the prevalence of the disease is 15% (table 5). Proven reduction in the prevalence of disease in the variant strain Century amyloliquefaciens VKPM B-10642 - 1.6 times, with strain Century licheniformis VKPM B-10562 - 1.5 times.

Table 5
The influence of bacterial strains of the drug No. 3 defeat raspberry leaf neighbour (%SHA "Gardens of Siberia", accounting, July 28)
OptionsThe prevalence worksheets, %The extent of the lesion, %
Control (without treatment)15,010,0
.amyloliquefaciens VKPM B-106429,3*8,8
.subtilis ITCM IN-1064111,0to 12.0
.licheniformis VKPM B-1056210,0*8,0
The drug is No. 3 on the basis of a mixture of strainsthe 10.1*9,0
NDS054,6Ff<F05

Lat is her during August the disease in all cases there was little or no dry, relatively cool weather. Against this background, at 2-m accounting (final) - August 21, trend lesion was shown to be less pronounced and statistically was not confirmed (table 6). In General, the defeat of Septoria, including in the control variant was weak and had no harmful effect.

Table 6
The influence of bacterial strains and preparation # 2 defeat raspberry leaf neighbour (%SHA "Gardens of Siberia", accounting, August 21)
OptionsThe prevalence worksheets, %The extent of the lesion, %
Control(without treatment)of 17.011,0
.amyloliquefaciens VKPM B-10642the 11.66,8
.subtilis ITCM IN-1064112,26,2
.licheniformis VKPM B-1056213,011,0
The drug is No. 2 on the basis of the e mixture of strains 12,38,0
NDS05Ff<F04Ff<F05

The incidence of purple blotch (caused by Didymella applanata (Nessl) Sacc., Dothideales, Ascomycota) in the experiment was slightly higher than the neighbour. The first symptoms were detected during the laying of experience 15 July. The growth of lesions in the control began with the end of the 3rd decade of July, in the advanced options in August. As a result, when accounting on August 21 in the control of disease prevalence amounted to 44.0%, the development is 12.5% (table 7). Significant (P<0.05) reduction in the prevalence of purple spot was detected in the variants with the strain .subtilis ITCM IN-10641 (1.6 times, EB=36,4%) and strain .licheniformis VKPM B-10562 (2.0 times, EB=50,0%). A significant reduction of lesion 1.8 times confirmed in the variant strain .licheniformis VKPM B-10562 also in terms of developing the disease.

As a result of a series of experiments (field, model, production) revealed a General picture of the effects from the use of bacterial strains in different cultures.

Table 7
The influence of bacterial strains and preparation # 2 on time is giving raspberry stems purple blotch (%, The SHA "Gardens of Siberia", accounting, August 21)
OptionsDisease prevalence, %EB, %The development of the disease, %
Control44,012,5
.amyloliquefaciens VKPM B-1064236,018,210,0
.subtilis ITCM IN-1064128,0*36,49,0
.licheniformis VKPM B-1056222,0*50,07,0*
Preparation No. 2 (mixed strains)28,734,98,7
NDS0515,7-the 4.7

Strains .amyloliquefaciens VKPM B-10642, .subtilis ITCM IN-10641, .licheniformis VKPM B-10561, .licheniformis VKPM B-10562 drug # 3 was reduced defeat raspberry different shot hole caused by fungal pathogens, in particular purple spots.

Example 6

With the aim of protection the berry crops during the growing season from fungal diseases conducted twice the processing of black currant cultivars Galinka water suspension .licheniformis VKPM B-10561; .licheniformis VKPM B-10562; .subtilis VKPM B-10641; .amyloliquefaciens VKPM B-10642 drug No. 3 against Septoria. As control was used plants treated with water.

Conducted field tests of all the studied strains against the infestation by Septoria currant showed that the disease has decreased 2.5-fold compared with control - 15% on July 10.

Surveys during the growing period show that the development of the disease in the variants with strains did not exceed 15% in the middle of the growing period and ranged from 18 to 20% at the end of the vegetation period, however, in the control of plants are affected reached 27%. At the end of the third decade of August the incidence of Septoria when using biologics does not exceed 40%and in the control reached 55%.

Biological efficacy of strains Century licheniformis - PMBC-10561 and VKPM B-10562 on July 10, was 68% and 79%, respectively, under the influence of the strain Bacillus subtilis VKPM B-10641 about 85%, a Bacillus amyloliquefaciens VKPM B-10642 - 70%. To July 17 biological efficacy of Bacillus licheniformis In PMBC-10561, VKPM B-10562 was the same and amounted to 67%. Biological efficacy of B. subtilis VKPM B-10641 was 77%, and Century amyloliquefaciens VKPM B-10642 accounted for 67 - 68%.

Example 7

The positive influence of strains .amyloliquefaciens VKPM B-10642, B. licheniformis VKPM B-10562, .subtilis ITCM IN-10641 drug No. 1 when landing on rooting travelnews the cuttings of black currant. The dates of planting cuttings of currants in the experience : 2nd decade of may. The concentration of the applied strains in the advanced options is 105CFU/ml Number of cultivated plants - 700 pieces 1 option. The method of application of bioagents - spraying the soil surface with the working fluid containing bioagent with subsequent moistening the soil to a depth of 10-15 cm by irrigation water. The hydraulic fluid option 1 - 10 litres.

Action test bacterial strains of the drug No. 3, become rooted cuttings of black currant were performed on plant varieties Xenia.

During the growing season were used to monitor the condition of the plants, which did not reveal any malicious background infectious diseases and pests. Final evaluation of the condition of the plants held at the end of the growing season on September 25. With this estimate, the plants were dug up, they weighed and measured.

To control the length of the vegetative growth of the main shoot was $ 14.4 cm (table 8). Under the influence of treatments strains Century amyloliquefaciens VKPM B-10642 and B. licheniformis In PMBC-10562 significant differences in this parameter was not detected. Under the influence of the strain B. subtilis ITCM IN-10641 increase significantly (P<0,05) increased by 4.5 cm (1.4 times).

Table 8
The effect is the processing of bacterial strains of drug # 1 on the growth parameters and biomass woody cuttings of black currant (sort Ksyusha, kennel SHA "Gardens of Siberia", accounting, September 25)
OptionLength of annual growth, cmThe length of the root system, cmBiomass of 1 plant
g% of control
Control (without treatment)14.4V20,712,1-
.amyloliquefaciens VKPM B-1064214,523,8*13,711,9
.subtilis ITCM IN-1064119,9*24,9*18,8*54,1
.licheniformis VKPM B-1056215,423,5*14,115,2
The drug is No. 1 on the basis of a mixture of strains16,624,1*15,527,1
NDS052,1 1,83,4

The length of the root system of the seedlings in the control averaged 20.7 cm, under the influence of treatments strains length of roots was significantly increased. In particular, under the action of strain .amyloliquefaciens VKPM B-10642 - 3.1 cm (15%), under the effect of strain .subtilis ITCM IN-10641 - 4.2 cm (20%), under the effect of strain .licheniformis VKPM B-10562 - 2.8 cm (14%). In the control variant, the average biomass of 1 plant was 12.1, In the experimental variants were observed trend of increasing biomass under the influence of the treatments.

However, reliably increased biomass relative to control only under the action of a strain of B. subtilis ITCM IN-10641 - 6.7 g (54.1 per cent).

Thus, the greatest effect on the effect on plants of black currant in the process of woody cuttings rooting was achieved when using a strain of B. subtilis ITCM IN-10641 preparation No. 1.

Example 8

Processing black currant directed against Septoria and Anthracnose. Processing single - when the first symptoms of the disease. The concentration of the used strains of drug No. 3 in the advanced options is 106CFU/ml. the Method of application of bioagents - ground mechanized spraying (MT3-80+OECS-2200). The flow rate of the working fluid at the rate of 500 liters per 1 ha - 125 l option.

In production the experience to test the validity of bacterial strains to defeat black currant fungal diseases treatments were conducted on July 15, 2010 The drug was applied to the plants tractor sprayer OECS-2200.

At the time of processing plants currant single spot lesions Anthracnose (caused by Gloeosporium ribis (Lib.) Mont, et Desm.., Melanconiales, Deuteromycota) was observed in 3-5% of the total number of leaves. To date 1 accounting - July 28, extent of the lesion in the control reached 14.5%, prevalence of disease and 17.6% (table 9). The prevalence of the disease in this period there has been a downward trend in this indicator 1.4 - 1.5 times as compared with control. On the extent of damage Anthracnose differences 2 weeks after spraying showed significantly (P<0,05): the degree of the damage of the leaves under the influence of strain Century amyloliquefaciens VKPM B-10642 decreased 3.2 times that of B. subtilis ITCM IN-10641 - 2.3 times, .licheniformis VKPM B-10562 - 2.2 times.

Until the 3rd decade of August there occurred an increase in the incidence of Anthracnose in the control variant 2 times up to the level of 34.6%. Differences from control in the end proved to be significant (P<0,05) for all applied strains: under the influence of strain .amyloliquefaciens VKPM B-10642 the incidence of Anthracnose was reduced by 3.8 times, under the influence of strain .subtilis ITCM IN-10641 - 1.6 times, under the influence of strain .licheniformis VKPM B-10562 - 2.3 times.

Table 9
The influence of bacter the social strains of drug # 2 on the defeat of black currant leaves the Anthracnose and Septoria leaf blotch (SHA "Gardens of Siberia")
OptionsAnthracnose, %Septoria, %
the prevalence worksheetsthe degree of destructionEB, %the prevalence worksheetsthe degree of destructionEB, %
July 28August 21July 28August 21August 21August 21
Control17,634,614,523,3-23,0of 17.0-
.amyloliquefaciens VKPM B-1064211,99,2*4,6*6,0*74,215,26,* 61,2
.subtilis ITCM IN-1064112,921,6*6,3*13,2*43,316,09,2*45,9
.licheniformis VKPM B-10562the 11.615,2*6,5*7,8*66,5of 17.010,0*41,2
The drug is No. 2 on the basis of a mixture of strains12,115,3*5,8*9,0*61,316,18,6*49,4
NDS05options9,34,8-Ff<F055,0-
NDS05timing6,6 3,4----

Similarly showed differences with control and indicator of the extent of disease. To control the degree of the damage was the result of 23.3 per cent. Under the influence of strain Century amyloliquefaciens VKPM B-10642 this figure was significantly decreased by 3.9 times (EB=74%), under the influence of the strain B. subtilis ITCM IN-10641 - 1.8 times (EB=43%), under the influence of strain Century licheniformis VKPM B-10562 - 3.0 times (EB=67%).

The defeat leaves a white spot (Septoria leaf blotch, caused by Septoria ribis Desm.., Spaeropsidales, Deuteromycota) was discovered in early August. By August 21, the disease spread to all grown at the experimental station of the plant, affecting them 23,0% leaves. In the variants with the use of bacterial strains, there was a decreasing trend in the prevalence of the disease in the 1.6 - 1.7 times, and in terms of the extent of damage the effect was proved in all 3 strains. Under the influence of the processing strain Century amyloliquefaciens PMBC In 10642 degree Septoria lesions decreased 2.6 times (EB=61%), B. subtilis ITCM IN-10641 - 1.8 times (EB=46%), B. licheniformis VKPM B-10562 - 1.7 times (EB=41%). Thus, against Septoria and, in General, shot hole leaf black currant greatest efficiency was handling the strain Century licheniformis VKPM B-10562.

In provideno the series of experiments (field, model, production) revealed a General picture of the effects from the use of bacterial strains of the drug №3 different cultures: the strain Century amyloliquefaciens VKPM B-10642 had a pronounced effect of the growth of the roots of black currant. In addition, this strain has reduced the defeat of the various cultures of the shot hole caused by fungal pathogens; the strain of B. subtilis ITCM IN-10641 increased biomass and length of the roots of black currant, increased the length of the annual increment of above-ground system of the black currant, reduced the incidence of all culture-shot hole leaf; strain Century licheniformis VKPM B-10561 caused an increase in the length of roots in cuttings of black currant, effectively inhibited the development of mushroom shot hole black currant.

A positive treatment effect of bacterial strains and preparation No. 3 on the basis of these strains on growth parameters and biomass berry crops are presented in table 10.

Table 10
The treatment effect of bacterial strains and preparation No. 3 on the basis of these strains on growth parameters and biomass berry plants
OptionLength of annual growth, cmThe length of the root of the second system, cmBiomass of 1 plant
g% of control
Control (without treatment)15,122,0to 12.0-
.amyloliquefaciens VKPM B-1064214,224,7*13,911,5
.subtilis ITCM IN-1064116,8*24,9*18,4*15,3
.licheniformis VKPM B-1056215,024,8*16,013,3
.licheniformis VKPM B-1056116,024,6*16,213,5
The drug is No. 3 on the basis of a mixture of strains15,524,716,1the 13.4
NDS0514,320,2 to 12.0-

1. The way to stimulate growth and protect fruit crops from diseases caused by fungal pathogens, including processing plants, biological product in liquid form, wherein the plant is treated by soaking the roots of the seedlings before planting, or spray them after planting, or spray plants during the growing season and fruiting, or spray the soil around the plants with subsequent moistening with water, and as a biological product, a mixture of bacterial strains .subtilis VKPM B-10641, .amyloliquefaciens VKPM B-10642, .licheniformis VKPM B-10561 and .licheniformis VKPM B-10562, or a mixture of strains of bacteria .subtilis VKPM B-10641, .amyloliquefaciens VKPM B-10642 and .licheniformis VKPM B-10562 with a title for each strain of not less than 105CFU/ml in aqueous suspension.

2. The method according to claim 1, characterized in that the stimulation of the growth and protection of strawberries from diseases caused by fungal pathogens, her landing in the soil is carried out after soaking the roots of the seedlings in the aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: strain .licheniformis VKPM B-10562, strain .subtilis VKPM B-10641, strain .amyloliquefaciens VKPM B-10642 with a title for each strain equal to 105CFU/ml, and fruiting strawberry plants are sprayed with an aqueous suspension of the drug containing a mixture in equal proportions of strains bacter the nd: .licheniformis VKPM B-10562, .subtilis VKPM B-10641, .amyloliquefaciens VKPM B-10642 with a title for each strain equal to 106CFU/ml

3. The method according to claim 1, characterized in that the stimulation of the growth of buckthorn seedlings sprayed the soil between the plants with an aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: .licheniformis VKPM B-10562, .subtilis VKPM B-10641, .amyloliquefaciens VKPM B-10642 with a title for each strain equal to 106CFU/ml, followed by moistening the soil to a depth of 10-15 cm irrigation water.

4. The method according to claim 1, characterized in that the protection planting raspberries or blackcurrants from diseases caused by fungal pathogens, landing sprayed with an aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: .licheniformis VKPM B-10561, .licheniformis VKPM B-10562, .subtilis VKPM B-10641, .amyloliquefaciens VKPM B-10642 with a title for each strain equal to 106CFU/ml with a flow rate of the aqueous suspension of 500 l per 1 hectare

5. The method according to claim 1, characterized in that the stimulation of seedling growth of black currant sprayed the soil between the plants with an aqueous suspension of the drug containing a mixture in equal proportions of bacterial strains: .licheniformis VKPM B-10561, .licheniformis VKPM B-10562, .subtilis VKPM B-10641, .amyloliquefaciens VKPM B-10642 with a title for each strain equal to 106CFU/ml with subsequent moistening soil between the plants to a depth of 10-15 cm irrigation water.



 

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