Water-dispersed composition for delivery of fungi designed for biocontrol, reducing content of aflatoxin

FIELD: biotechnologies.

SUBSTANCE: composition contains an agent designed for biocontrol, for reduction of level of pollution of food and fodder with aflatoxin, a binding agent, an agent having osmoprotector and adhesive properties, a carrier and a source of nutrient elements for an agent, designed for biocontrol.

EFFECT: reduced level of food and fodder contamination with aflatoxin in raw materials.

29 cl, 4 tbl, 6 ex

 

The technical field

The present invention relates to water dispersible granular composition, conducting biological control, designed to reduce aflatoxin contamination of food and feed for animals, in particular, corn, and also to methods of preparing such composition. Water dispersible granular composition contains an agent that is designed to biocontrol, enclosed in a granular matrix, forming a dispersion by adding water. Agent intended for biocontrol, are netoxygen and neoplatonically strains of Aspergillus flavus, is able to inhibit the formation of colonies aflatoxinogenic mushrooms, and also able to inhibit the production of aflatoxin toxicogenic fungi. Water dispersible granular composition according to the invention has a high stability during storage and in the field.

The technical field

Many fungi produce secondary metabolites that are not necessary for their growth and reproduction. If these metabolites are toxic to humans or animals, then they are classified as mycotoxins. There are four most important genus of fungi that produce mycotoxins, including: Aspergillus, Fusarium, Penicillium and Alternaria (Council for Agricultural Science and Technology [CAST]. 2003. Task Force Report 139, Ames, IA). These fungi produce Miko is oxine, which can adversely affect the quality and delivery of food and feed, including corn, cotton seeds, cereal grains, peanuts and tree nuts.

The total amount of damage caused by mycotoxins in the food industry and animal husbandry in the USA and Canada is estimated at five billion annually; however, the greatest concern is aflatoxin - class of mycotoxins produced by strains of Aspergillus spp. (Robbens and Cardwell. 2005. In: aflatoxin on and Food Safety, Abbas, H.K. (Ed.), CRC Press, Boca Raton, FL, pp.1-12). The two main types of mycotoxins, prevailing as contaminants of food and feed are produced A.Flavus mycotoxins - aflatoxins B1 and B2 (Payne, G.S. 1992. Critical Rev. Plant Sci. 10: 423-440). I believe that aflatoxin Bi (AFB1) is the strongest and most common mycotoxin (International Agency for Research on Cancer-World Health Organization [IRAC-WHO]. 1993. In: IARC Monographs on the Evaluation of Carcinogenic Risks to Humans, Lyon, France, pp.56, 467-488). The contamination is most often associated with A.flavus (Diener et al. 1987. Ann. Rev. Phytopath. 25: 249-270). This fungus is able to grow in a wide temperature range, namely 10°C-43°C, and a wide range of water activity (0.82-0.998) (Food and Agriculture Organization of the United Nations/ International Atomic Energy Agency [FAO/IAEA]. 2001. In: FAO Food and Nutrition Paper, FAO, Rome, Italy, pp.73, 75-93). However, dry weather, mechanical damage or the presence of pests tend to increase zagryaznennost the ü corn with aflatoxin in the pre-harvest period.

Currently, the maximum allowable levels of aflatoxin in human food and animal feed is 20 µg/kg (CAST, supra; van Egmond and Jonker. 2004. J.Toxicol. - Toxin Rev. 23: 273-293). Despite the fact that to avoid the presence of mycotoxins in agricultural commodities is not possible, the level of contamination can be controlled with the use of certain agricultural practices. Some variants of pre-harvest strategies varying degrees of success, regulating aflatoxin contamination (Betran and Isakeit. 2004. Agron. J. 96: 565-570). A promising control strategy is biological control using netoxygen A.flavus (Domer, J.W. 2004. J.Toxicol. - Toxin Rev. 23: 425-450). Brown et al. (1991. J.Food Protect. 54: 623-626) showed that the levels of aflatoxin can be reduced by direct scarification corn cobs and injection netoxygen strains of A.Flavus. In contrast to the direct, mechanistic strategy Brown et al., did the strategy of indirect delivery in soil that received more widespread. Soil inoculum, as a rule, is an active, netoxygen strain .flavus who originally cultivated grains of cereals. These grains are nutrient medium providing the proliferation of strain .Flavus intended for biocontrol, and their use for the treatment of soil under the planned grain the cultures. While on grains, netoxygen strain produces many spores spread by wind and water, and competes with endemic aflatoxigenic strains for resources, which in General leads to lower levels of aflatoxin. This strategy tillage has been used successfully for growing peanuts (Domer et al. 1992. J. Food Protect. 55: 888-892), cotton (Cotty, P.J.1994. Phytopath. S4: 1270-1277) and maize (Domer et al. 1999. J. Food Protection 62: 650-656). A similar strategy using inoculum applied to soil, used in the production of corn in the Mississippi Delta (Abbas et al. 2006. Biocontrol Sci. Tech. 16: 437-449). Study of the process of production of corn in the Mississippi Delta revealed C - netoxygen strain of A. flavus, which contributed to a significant reduction in aflatoxin contamination during four years of field trials and showed a high capacity to colonize.

Despite the success of the above strategies, there are obstacles that limit their practical application in commercial agricultural production. Consequently, there remains a need to develop approaches involving direct air delivery, allowing you to achieve an effective reduction of the level of contamination of maize with aflatoxin in the pre-harvest period and to reduce the dependency of applications from optimal environmental conditions with the food.

BRIEF description of the INVENTION

The authors of the present invention is designed composition, representing a water dispersible granular composition comprising biocontrol agents suitable for use as a sprayable conidial suspension designed to prevent aflatoxin contamination of food and feed, as well as the method of preparation of the water dispersible granular composition.

Thus, in accordance with the foregoing, the present invention is the provision of water dispersible granular composition containing isolated neoplecostominae and netoxygen strains of A. Flavus, can function as agents of biocontrol and to inhibit the proliferation of the fungi that produce aflatoxin, thereby preventing the contamination with aflatoxin food and feed. According to preferred examples of implementation of the proposed netoxygen strain, designated as C.

In addition, another objective of this invention is the provision of such a composition on the basis of control agents, in which these agents would preserve the viability, with a high degree of stability of the composition during storage and in the field.

Another object of the invention is the preparation of products for the unit is ntrolle, that are clean, easy to handle and has a relatively low phytotoxicity to crops.

Another object of the invention is the inclusion of biocontrol agents in the compositions suitable for use in traditional agricultural sprayers.

Other objectives and advantages of the present invention will become apparent from further description.

DETAILED description of the INVENTION

Autorama of the present invention, a method of preparing water-dispersible granular composition containing the biocontrol agent - netoxygen strain of A. flavus. Compared the effect of inoculation of soil netoxygen strain K included and not included in the composition, to determine the impact on the ability to colonize and levels of aflatoxin in field corn. This comparison was performed with the aim of orankestra direct delivery to reduce aflatoxin contamination of corn in the pre-harvest period. A similar degree of colonization and reduce the level of aflatoxin was detected when comparing the effect obtained by spraying conidia included and not included in the composition. The value of the detected data is that they demonstrate the ability of the product development of biological control in use is lovanii traditional application techniques, in order to reduce contamination of maize with aflatoxin. Excellent result, which characterizes the decrease in the levels of aflatoxin, and the apparent survival rate of strain intended for biocontrol, supports the hypothesis that the most effective way to reduce the level of contamination of maize with aflatoxin, is a direct application of the biocontrol agent to susceptible to aflatoxin or reproductive organs of maize.

The method consists in adding competitive, netoxygen strains of A. Flaws in the soil, used in the traditional way; the result has been a decrease in the concentration of toxins in crops, due to the fact that biological competition netoxygen strains of Aspergillus with the microflora of the soil prevented the accumulation of strains producing toxins, usually appearing during the late seasonal droughts. During the competitive replacement of toxicogenic strains of fungi living in soil under natural conditions, was replaced methoxycinnamic and neoplachennyj strains added to the soil. Therefore, all crops affected by late seasonal drought was amazed prevailing bioconcentrate strains are unable to produce toxins.

However, the use of grain inoculants to regulate the possession of aflatoxin in maize is associated with the following drawbacks: 1) the use of the solid matrix may be difficult for commercial use in the case when grains are in the later stages of ontogenesis, 2) some of the biotic, abiotic and weather factors may limit or stop the dispersal of conidia from granular sources to the organs of maize available for air delivery 3) the sporulation occurring on the treated grains in the field may cause problems related to health and safety. In the end, the success of the strategy of biological control depends on the dynamics of biological competition between different strains of A. flavus. Conidia K included and not included in the composition, was applied on the reproductive tissues of maize and compared their effects on reducing the content of aflatoxin in corn. When directly sprayed on corn on the cob is included and not included in the composition conidia were highly effective in reducing the level of contamination of maize with aflatoxin. Despite the fact that the material included in the composition, prepared and kept for eleven months, with no difference in effectiveness between the material included in the composition, and a material consisting of freshly harvested conidia are not included in the composition. Since the application svezhevyrabotannym biocontrol agents is a difficult condition from a commercial point the of view, stable composition capable of effectively delivering the biocontrol agents, is an important and commercially feasible product that allows you to adjust the levels of aflatoxin in corn and other crops, are susceptible to mycotoxin contamination. The concentration of material used is also different. In this research material included in the composition, the use of 9 kg/ha; while the grains of soil inoculants are traditionally applied in an amount of from 20 to 200 kg/ha (Abbas et al. 2006; Cotty; Domer et al. 1998; supra). Further optimization of the composition described herein, as well as atomization method may further contribute to the reduction of the quantity of composition applied per unit area, for regulating the content of aflatoxin in corn.

The method according to the invention is applicable to any agricultural raw materials cultivated for human consumption and/or as feed for animals and/or damaged by the toxins of fungi for which direct application to the target organs of plants, such as maize, cotton, hazelnut and olive, has a beneficial effect.

In accordance with the objectives of the present invention, the term "drug mushrooms or agricultural composition, prednaznachennaya for biocontrol containing mushrooms" Rel which relates to microbial drug in which microbacterial, essentially consist of, or are netoxygen or neoplecostominae strains of Aspergillus. Drugs mushrooms can contain one or more netoxygen or neoplastigenic strains of Aspergillus. "Netoxygen strain Aspergillus)) includes any strain that does not produce the toxin aflatoxin and cyclopiazonic acid (CPA). "Neoplecostominae strain Aspergillus" includes any strain that does not produce the toxin aflatoxin, however, continues to produce cyclopiazonic acid (CPA). Agricultural composition intended for biocontrol containing mushrooms, in accordance with the objectives of the present invention, includes netoxygen strain (or strains) of mushrooms, or Neoplecostominae strain (or strains) of mushrooms, enclosed in an acceptable agricultural carriers, which can be any media that you can attach mushrooms and which are not harmful fungi or grain, which is treated with a composition. Example netoxygen strain includes A. flavus K49. Mushrooms, particularly useful when applied in accordance with the present invention, represented by strains with identification characteristics netoxygen A. flavus K49, designated as NRRL 30797. These characteristics are the inability to production of toxin aflata the Cena and CPA, as well as the ability to biological competition in the soil, which produces an agricultural commodity culture. Example neoplecostominae strain includes A. flavus CT3. In addition, fungi, particularly useful when applied in accordance with the present invention, represented by strains with identification characteristics neoplecostominae strain A. flavus CT3, designated as NRRL 30798. These characteristics are the inability to producing aflatoxin, as well as the ability to biological competition in the soil, which produces an agricultural commodity culture.

When netoxygen and neoplecostominae strains of Aspergillus cultivated as single strains in granular food sources, such as, for example, wheat, rice, rye, etc., these food sources with full colonization contain approximately 108colony forming units (CFU) of fungi per gram weight of the food source. These food sources inoculated grain incubated at approximately 35°C. After 24 h growth of inoculated wheat were shaken manually and incubated for 24 h in addition, further homogenized by shaking manually. Colonization by strain inoculant confirmed by determining the concentration of aflatoxin in inoculants. Inoculated p is oduct can be stored at approximately 5°C for approximately 2 months or more if the water content in the dried product below critical.

Netoxygen and neoplecostominae strains of Aspergillus applied to plants in amounts effective to reduce the levels of toxins in agricultural raw materials. The term "lower levels of toxin" in the present description refers to a decrease in the number of toxin compared with the amount of toxin in agricultural raw materials, expected in the case where the agricultural raw materials are not processed by the methods according to the present invention. For the person skilled in the art it is obvious that for such comparisons, you can apply all accurate methods of measuring and comparing levels of toxin.

The terms "effective amount", "amount effective", "effective amount" refers to that introduced the number one drug of fungi, in which the effect of its use is manifested in the reduction of toxin contamination of agricultural raw materials.

EXAMPLES

After a General description of the invention for a better understanding presents specific examples are included for purposes of illustration and not limiting of the present invention, the scope of which is defined by the claims.

EXAMPLE 1

Strains of Aspergillus flavus

Netoxygen strain A. flavus K49 (NRRL 30797) and aflatoxinogenic F3W4 (NRRL 30796) kept on silica gel at 4°C, before the study examined the relevant phenotypic characteristics of the strains, the profiles of aflatoxin, the formation of sclerotia, the morphology of the colonies and the formation of conidia (Abbas et al. 2006. Biocontrol Science and Technology 16: 437-449).

EXAMPLE 2

Materials composition and water-dispersible granular preparation

As a carrier in the preparation of water dispersible granular composition (WG) used soda Kalinovo clay with an average particle size less than 1 micron. In addition to trehalose as a binder used sodium carboxymethylcellulose. Trehalose was used as a multi-component composition. This disaccharide was included as osmoprotective, adhesive or the adhesive functioning after application, and a potential source of nutrients for K. In particular, the composition of the dry ingredients in the composition contained:

76-90% Satintone 5HB as a carrier, 1-4% Nilyn XL 90 as a binder, and a 5-20% trehalose as osmoprotective, adhesive or the adhesive functioning after application, and a potential source of nutrients for K. The dry ingredients were mixed until visual homogeneity in the mixer with large shear effort before mixing in approximately 510 ml of 0.1% (VASAB) peptone solution, containing 5% of total dry trehalose and conidia K at 4×108CFU/g wet mixture of 500 g of the dry ingredients. Conidia were harvested from agar cups with the best extract by adding a small aliquot of 0.1% peptone solution. Thus, conidia were prisoners in the pellets made from the composition. Control pellets not containing conidia of A. flavus, were prepared and processed as described above.

Clay, other than calcined kaolin clay, can be used in the compositions according to the invention, i.e. compounds you can use any clay with an appropriate particle size, i.e. a particle size comparable with the size of body, small enough not to cause clogging of spray. Thus, it is possible to use other siliceous clays and mixtures of clays, such as bentonite, kaolinite and smectites, including montmorillonite and beidellite.

The above mixture was prepared separately using a disc pelletizer (LCI Corp) matrix 1.2 mm or 2.0 mm, was dried under vacuum until the water activity, which constitutes approximately 0,30. Granules with a size of 2.0 mm and 1.2 mm is designated as Product 1 and Product 2. respectively. Pellets were stored at 4°C for ~330 days; during this period, the survival of propagules of A. Flavus were periodically measured sowing on pruselect the ing the environment. Three parallel samples homogenized in water agar (0.2% wt./about.) reciprocating shaking (30 min, 100 beats/min), were serially diluted and were sown on a modified environment with Bengal rose (MDRB; Horn and Domer. 1998. Mycologia 90: 767-776).

Analysis of the granulated composition comprising conidia are carried out immediately after drying, showed >3×108CFU/hotmetal relatively high survival of A. flavus strain K, which indicates no loss of viability of the strain within 11 months of storage C included in the composition at 4°C (table 1). After two years of storage the number of viable propagules of fungi was reduced by only 50%.

Pellets of the composition contain a prisoner in their biocontrol agent, in this case, conidia C. Upon contact with water (as in Example 5) or other aqueous solution, the granules form a dispersion or dissolved with the release of the biocontrol agent who acquires the ability to function as a biocontrol agent.

Table 1
Survival A.flavus strains C in water-dispersible granules.
Product 1 (2.0 mm WG)Product 2 (1.2 mm WG)
Colony-forming units A.flavus 1 g
After drying3.12±0.35×1083.70±0.10×108
Storage at 4°C for 20 days2.57±0.15×1082.95±0.34×108
Storage at 4°C for 330 days3.90±0.36×83.18±0.21×108
Storage at 4°C for 745 days1.66±0.44×1081.49±0.20×108
Data presented are mean values of three samples ± standard deviation.

EXAMPLE 3

Conidia and solid preparation for inoculation

For inoculum production of conidia, are not included in the composition of the original culture were sown at 40 cups potato-dextrose agar (KDA), and incubated for 5-7 days at 28°C, under a 12 hour light regime (165 umol/m2/s1and 12-hour dark conditions. Conidia and mycelium was removed from the Cup using water tween-20 (0.2% wt./vol.). Vegetative fungal structures were separated from the suspension of conidia by filtration through DV is a layer of gauze. The density of conidia was measured using hemocytometer; final concentration is brought to a value of 4.1×106conidia/ml

For inoculation of the soil used wheat as a carrier inoculum, as described in the literature (Abbas et al. 2006, supra). Wheat soaked in water overnight, dried and placed in autoclave bags (1 kg/bag with 250 ml of water) for 1 hour at 121°C. the Primary inoculum of A. flavus was the 5-day culture KDA - section area of 3 cm2on the bag, which were incubated at 35°C. After 48 hours at 35°C wheat was fully colonized. Then this product is homogenized by manual shaking and kept at 4°C before use in field tests.

EXAMPLE 4

A field study of the colonization using the techniques of "Pin Bar" Method of inoculation with the use of technology "pin bar" (Windham et al. 2003. J.Toxicol. - Toxin Rev. 22: 313-325) was used to compare the degree of colonization of maize inoculated with a suspension of conidia C not included in the composition, and maize inoculated WG granulirovannykh composition comprising C in 2005 in the field trials conducted in Stoneville and Elizabeth, Mississippi. In Stoneville studies were performed on the hybrid maize resistant to glyphosphate (Garst 8270 rr), and in studies in Elisabeth participated hybrid expressing the gene is endotoxin Bacillus thuringiensis (Agrigold A6333 bt). Corn cobs were inoculable 25 days after 50% tasseling pistillate columns (development of dent grains). Corn cobs (100 per treatment) were inoculable individually suspension included or not included in the composition of conidia (5×106conidia/ml), in the phase of emergence pistillate columns in 50% of plants with the help of technology "pin bar" (three 100 mm number of 12 sewing needles, mounted on wooden block, the tip of each needle is 6 mm). The needles were dipped in a suspension of conidia and were immersed into the cob. Through various time intervals after the "pin bar" inoculation were collected from ten inoculated ears treated the same way; the total number of cobs in the inoculated area and the number of infected ears was determined by counting and visual assessment of fungal growth on separate ears.

Similar to the final level of colonization of corn cobs strain K introduced in the form of conidia, included or not included in the composition, observed in 2005 at two sites in the study with the "pin bar" inoculation (table 2). The initial speed of colonization of corn strain C observed at stations located in Stoneville (not-WATT hybrid) was higher compared with the speed at sites located in Elizabeth (WATTS hybrid). In studies in which onville inoculation was carried out on 10 days earlier than Elizabeth, and, possibly, on the rate of colonization affected by weather conditions. Cases of colonization A.flavus on the control pinakamaraming sites Stoneville observed more frequently than in Elizabeth. This difference is possibly caused by the use of WATTS hybrid (on sites located in Elizabeth), reducing the frequency of lesions corn borer European comborer (Dowd, .W. 2003. J.Toxicology-Toxin Review 22: 327-350). On the plot in the area of Elizabeth watched accelerated colonization of corn kernels on the cob, inoculated with Product 1, compared with colonization on the cob, inoculated with conidia that are not included in the composition, after 5 and 7 days after inoculation; at the same time, in the area of Stoneville both composition contributed to enhanced colonization compared with colonization by inoculation with conidia that are not included in the composition, 7 days after inoculation. Enhanced colonization by strain C was observed after application of the Product 1 in comparison with the Product 2 in the area of Stoneville after 5 days and in the area of Elizabeth 9 days after inoculation. Not noted the negative impact of the composition and of the same composition after storage for 20 days) on the colonization of conidia K; early research at certain points mentioned more active colonization of conidia K included in the composition, compared with conidia that are not included in the song. Based on these preliminary results for use in field testing of the second year of the experiment were chosen Product 1 (diameter 2.0 mm WG).

Table 2
Colonization of corn grains after incubation K caused in the composition of the water-dispersible granules, or in the form of free conidia in two areas.
Infected grain (%)
Day 2Day 5Day 7Day 9Day 12Day 14
Elizabeth (W)*
Control000.82.1 b5.0 b7.5 b
Product 1054.1 85.7100.0100.098.1 and
Product 2051.173.197.2100.098.1 and
Free conidia034.4 b67.6 b100.0100.092.0
LSD LSB (Pr>0.05)16.210.04.58.210.8
Stoneville (RR)#
Control0.0 b0.95.19.4 b6.9 b23.9 b
Product 10.6 b 80.198.7 and93.898.599.2
Product 20.0 b37.5 b98.994.596.7 and93.6
Free conidia17.2 a69.785.7 b91.196.096.6 and
14.5512.08.95.214.3

Presents the average result of three samples collected every day. Values with the same letter indexes do not differ significantly at confidence level 95%. W=hybrid containing W design, RR=modified hybrid resistant to glyphosphate.

EXAMPLE 5

A field experiment on the regulation of the content of aflatoxin and implementation strains

In 2006, in the district of Stoneville, Mississippi, conducted a field study to assess the effectiveness of the NAS is the basis of toxicogenic strain K - the biocontrol agent on the leaves of the plant to reduce contamination aflatoxigenic isolates of A.flavus and contamination by aflatoxin. Corn (OK C69-70RR) were sown on April 14, 2006 in silty-clay soil (Dundee) Stoneville, Mississippi. Used an experimental randomized layout of the plot for 6 treatments, replicated in 3 blocks. Each experimental block consisted of three rows of length 9,36 m (1.06 m wide) with one processed next (in the centre from 25 to 30 plants) and two untreated buffer rows. Treatments included: 1) control - no treatment; 2) soil inoculum - wheat inoculated aflatoxigenic A. flavus strain F3W4 (20 kg/ha); 3) soil inoculum - wheat inoculated composition comprising C; 4) soil inoculum - wheat inoculated with both strains F3W4 and C; 5) processing the slurry extruded granules, including K; and 6) treatment sprayed with a suspension of freshly harvested conidia C.

26 June (early tasseling pistillate columns), wheat inoculant aflatoxigenic strain F3W4 and atoxigenic strain C has processed the corresponding plots of 20 kg/ha 30 June (at tasseling stage pistillate columns in 50% of plants), using a hand held sprayer sprayed inoculants (quenching and the TKA ways 5 and 6). The solution for spraying included 56 g of the product containing the composition, suspended in 4 liters of 0.2% wt./about. Tween-20, the number of working solution was ~600 ml per plot. Spent spraying the upper third of the plant, the target was the cobs. At the stage of physiological maturity (August 17) all the cobs from the plants, located 6 m from the centre of the treated series were collected manually, dried, thrashed them and grind.

The calculation of the density of propagules of Aspergillus and evaluation of characteristics of the isolate was carried out using a selective medium. Ground grain samples homogenized in 0.2% wt./about. water agar, diluted series and was inoculated on the MDRB agar. Colony-forming units (CFU) were counted after 5 days of incubation. Thirty colonies plot moved on CD-PDA (PDA with 0.3% β-cyclodextrin), and incubated for 5 days under continuous dark at 28°C, and assessed the production of aflatoxin by pigmentation of the colonies and the color change after exposure to vapors of ammonia (Abbas et al. 2004. Canad. J.Microbiol. 50: 193-199).

All field data, the results of colonization, excretion of toxin and phenotype Aspergillus and aflatoxin contamination analizirovali using PROC GLM of the Statistical Analysis System (SAS. 2001. SAS® Proprietary Software Release 8.2, Windows version 5.1.2600. SAS Institute Inc. Cary, NC). The separation medium was carried out according to the criterion of least significant once the spine (Fisher test).

The number of CFU of A. flavus isolated from milled grains ranged from log 5.4 to 6.3 propagules/g of grain at maximum separation from grain treated with a spray solution of conidia C not included in the composition (table 3). This level of colonization is much higher compared to the level detected during the counting propagules of A. flavus, found on maize in previous studies (Abbas et al. 2006, supra), where the number of selected colonies ranged from 3.4 to 4.4 log CFU/g of grain. The minimum share (from 4 to 6%) aflatoxigenic isolates found in the application of sprayed strain K included and not included in the composition (table 3). When all other methods of treatment have observed similar levels aflatoxigenic isolates (from 50%to 71%), which was not significantly affected by inoculation of soil as netoxygen strain K and toxicogenic strain F3W4.

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Table 3
The selection of isolates of A. flavus from maize under different inoculation processing.
ProcessingA. flavus (log10CFU/r)Toxicogenic isolates (%)
Without inoculation5.5 be
C wheat inoculant5.7 b52
F3W4 wheat inoculant5.7 b71
F3W4+C wheat inoculant5.6 bc50 and
F3W4 wheat inoculant +C composition for spraying, including conidia5.44 b
F3W4 wheat inoculant +C available conidia (not included in the composition)6.36 b
M3PLSD(Pr>0.05 level)0.242
Presents average values for the three experiments. Values with the same letter indexes do not differ significantly at confidence level 95%.

According to the average results of SOME calculations and the recording frequency of the two phenotypes was determined ratio toxicomanie:toxicogenic, amounting to 24:1, 15:1 and 1:1 for the spray solution containing the song; spray solution not containing compositions and all other processing methods, respectively. Previous research has shown that colonization by strain C corn cobs can be enhanced by inoculation of soil (Abbas et al. 2006, supra); however, this result was not confirmed in this study (table 3). A possible explanation for these differences is that in this study the soil inoculum K applied at tasseling stage pistillate bars at 50% of the plants, whereas in the previous study, the inoculum was applied at the V6 stage of ontogenesis. Studies on cotton Cony (supra) revealed that 67% were isolated from cotton bolls A. flavus were in the same group of vegetative compatibility, and netoxygen strain introduced in the form of soil inoculant; compared to 46% with the introduction of sprayable inoculant; and 25% on the untreated control plots. This indicates that under field trials in Arizona when applying by spraying conidia observed a lower degree of implementation biocontroller strain.

The low level allocation aflatoxinogenic A.flavus from corn, previous treatment by spraying conidia K included or not included in the composition shows a clear introduction, colonization and competitive replacement toxicogenic Asergillus embedded netoxygen strain K when directly applied on the reproductive patterns of maize.

EXAMPLE 6

Determination of aflatoxin and isolation of Aspergillus

The concentration of aflatoxin was quantified using commercial ELISA kits (Neogen Corp., Lansing, MI) according to Abbas et al. (2002, 2006, supra). Prepared three parallel hinge crushed grain (20 g); each was extracted with 100 ml of methanol (70%) for 30 min at high speed shaker for reciprocating movement, cleared by centrifugation (10 min, 8000×g) and analyzed methanolic extracts using ELISA. The limit of detection of aflatoxin in this study was 5 ng/g total aflatoxin.

In these field trials observed a relatively high level of aflatoxin contamination caused by natural infection in untreated control plots and control plots where the soil was inoculated with wheat grains infected aflatoxinogenic F3W4. Accordingly, the content of aflatoxin amounted to 428 and 635 mg/kg with a large scatter of results for the samples for these two control plots (table 4). However, where inoculation of soil toxicogenic K spent making the infected grains in areas that were either not treated, or treated soil inoculum F3W4, the content of aflatoxin decreased significantly (P<0.05) to 44 and 223 mg/kg, respectively. Here vnesenii soil C with F3W4 resulted in a significant decrease in the average levels of aflatoxin (~400 µg/kg) in corn compared with levels in the corn, inoculated only one F3W4. However, the levels of aflatoxin in maize inoculated with infected strains C with F3W4 grain was not significantly different from a similar level in corn with untreated control plots.

Table 4
The content of aflatoxin in corn treated with the use of different methods of inoculation
ProcessingThe concentration of aflatoxin (µg/kg)
Without inoculant428 ab
C inoculant (wheat)44
F3W4 inoculant (wheat)635
F3W4+K inoculant (wheat)223 be
F3W4 inoculant (wheat) +C sprayed composition18
F3W4 inoculant (wheat) +C available conidia (not included in the composition)21
LSB (LSD) (Pr>0.05 level)287
Presents average values for the three experiments, values with the same letter indexes do not differ significantly at confidence level 95%.

In previous studies, where K made in the soil, the contamination of maize with aflatoxin decreased by 58-76% compared to untreated plots, which are in abundance were present natural aflatoxin (Abbas et al. 2006, supra). Inoculation of soil aflatoxigenic isolate F3W4, simultaneous inoculation strain K on wheat was possible to reduce the pollution on 74-95% compared with the concentrations of aflatoxin in areas where the soil was inoculable only one F3W4. In other studies, the content of aflatoxin decreased by 66 and 87% for two consecutive years in the corn plots treated with equal mixtures of rice, colonized by two different neoplecostominae species of Aspergillus, in comparison with the content of aflatoxin on untreated plots (Domer et al. 1999, supra). Percent reduction of aflatoxin content was 90 and 65%; our results are consistent with results from previous studies. While the soil C led to a significant reduction of aflatoxin contamination, the concentration of aflatoxin was still exceeds the marginal rate of products, prednaznachen the th for use as a food raw material or feed for animals.

Processing corn by spraying with a suspension of conidia K included or not included in the composition, in areas where the soil was infected aflatoxinogenic F3W4, led to a decrease in the average concentration of aflatoxin at ~615 µg/kg, In particular, the direct application of conidia K included or not included in the composition, on the reproductive patterns of the corn grown in areas with soil F3W4, significantly (P<0.05) reduced levels of aflatoxin to 18 and 21 µg/kg, respectively, compared with levels of aflatoxin 635 mg/kg, detected in the control plot, where the soil is made only F3W4. Thus, the decrease in the average concentration of aflatoxin by more than 97% was associated with inoculation by spraying C, against 65% reduction in indirect inoculation with soil C. Experience with cotton making grains reduced aflatoxin contamination by 75%, at the same time when applying neoplecostominae strain by sputtering on reproductive tissues was not observed any effect (Cotty, supra). However, the aforementioned research on cotton were conducted in Arizona, where environmental factors (low relative humidity and high temperature) could impede the establishment of inoculant at air spraying.

Because of the anatomical differences between corn and cotton or ar is the Heath, inoculation of the soil may not be the most effective strategy for biocontrol of corn. Despite widespread use and apparent efficiency of solid inoculants for growing cotton and peanuts, this strategy of regulating the content of aflatoxin in corn is not adopted on a commercial scale. The difference of the experimental results with tillage and spraying can be explained by the difference method of application. Fruiting patterns peanuts are placed under the soil surface, therefore making inoculant in the soil aimed at the area of infection. In addition, the reproductive patterns of cotton is distributed from the third to the twelfth node and are relatively close to the soil surface. In contrast, maize is a fast-growing plant, a relatively high instances, produce sensitive reproductive patterns, situated at a height more than two meters from the soil surface. For colonization and subsequent replacement of natural aflatoxigenic strains on corn may require high levels of inoculum with the corresponding universal transfer mechanism incorporated into the soil biocontrol agents for introduction and establishment in the areas of target available for aerial spraying, ie, corn on the cob. At the same time, in sluchayah spray, this condition ceases to be necessary, as shown by this study. Another advantage of the direct spraying of the composition containing C is a commercial advantage - availability of ready available stable composition containing viable conidia C.

All publications and patents mentioned in this application is included in the application by reference, which is equivalent to the inclusion of each individual publication or patent separately and individually by reference.

The foregoing description and a separate representative examples of implementation and details of the present invention is provided for illustration and description of the invention. They are not exhaustive and do not limit the present invention is described forms. For specialists in this area it is obvious that within the framework of the present invention possible modifications and variations.

1. Water dispersible granular composition intended for biocontrol, containing: (1) the agent is intended for biocontrol to reduce the level of contamination of food and feed with aflatoxin; (2) a binder; (3) an agent having osmoprotective and adhesive properties; (4) the media and (5) the source of nutrients for agent intended for biocontrol in which the agent is intended for biocontrol, prisutstvuet is in the form of a suspension, the content agent with osmoprotective and adhesive properties, approximately 5-20% of the dry weight of the total composition, with these agents and the source of nutrients present in the specified composition in the form of a mixture, and the specified agent intended for biocontrol included in this mixture and when the dispersion in water is released with the ability to carry out its functions.

2. The composition according to claim 1, where the specified agent intended for biocontrol, is an effective amount of the drug conidia netoxygen or neoplecostominae strain of Aspergillus.

3. The composition according to claim 2, where the specified strain Aspergillus is selected from the group consisting of: Aspergillus oryzae, Aspergillus flavus, Aspergillus parasiticus, Aspergillus sojae, and mixtures thereof.

4. The composition according to claim 3, where the specified netoxygen strain Aspergillus is a strain K Aspergillus flavus.

5. The composition according to claim 1, where the specified binding agent is a sodium carboxymethylcellulose.

6. The composition according to claim 5, where the composition contains about 0.4% sodium carboxymethylcellulose.

7. The composition according to claim 1, where specified, the agent that has osmoprotective and adhesive properties, is a trehalose.

8. The composition according to claim 7, where the composition contains approximately 20% trehalose.

9. The composition according to claim 1, where the specified media pre which is a clay with particle size, comparable with the size of the conidia, small enough in order not to cause clogging of spray.

10. The composition according to claim 9, where the specified carrier is a silica clay or mixture of clays.

11. The composition of claim 10 where the specified silica clay or mixture of clays is a calcinated kaolin, bentonite, kaolinite or smectite.

12. The composition according to claim 11, where the specified media is a soda kaolin clay.

13. The composition according to item 12, where the composition contains approximately 75-90% of calcined kaolin clay.

14. The composition according to claim 11, where the specified smectite is a montmorillonite or beidellite.

15. The composition according to claim 1, where the specified source of nutrients is a trehalose.

16. The composition according to claim 1, where the composition is a dispersion suitable for spraying, can remain in suspension with a minimum of agitation/stirring.

17. The composition according to item 16, where this dispersion is suitable for spraying, is a water solution.

18. The method of preparation of the water dispersible granular composition intended for biocontrol, including agent intended for biocontrol, according to claim 1, which includes stages:
(a) mixing dry ingredients, including sodium carboxymethylcellulose, trehalose and flint is slightly clay or mixture of clays until visually homogeneous state;
(b) the suspension of an effective amount of conidia of Aspergillus netoxygen or neoplastigenic, peptone solution containing trehalose or other known osmoprotectant/nutrients for preparation of a solution containing conidia; and
(c) mixing the dry ingredients with a solution containing conidia, for the preparation of water dispersible granular composition intended for biocontrol, silica clay or mixture of clays is the media and
(d) vacuum drying,
where the sodium carboxymethylcellulose is a binding agent, trehalose is an agent with osmoprotective and adhesive properties, the source of nutrients and a binder agent, silica clay or mixture of clays is a media and nontoxigenic or neoplatonically Aspergillus conidia are an agent, intended for biocontrol.

19. The method of preparation of the water dispersible granular composition intended for biocontrol, including agent intended for biocontrol, p, where silica clay or mixture of clays includes calcinated kaolin, bentonite, kaolinite or smectite.

20. The method of preparation of the water dispersible granular composition intended for biocontrol, including Agay is t, designed for biocontrol, p, where silica clay or mixture of clays is a soda kaolin clay.

21. The method of preparation of the water dispersible granular composition intended for biocontrol, including agent intended for biocontrol, p, where the method of vacuum drying to produce granules.

22. The method according to item 21, where the granules produced by the method of vacuum drying, re-dispersed in water to obtain a composition suitable for spraying.

23. The method according to item 22, where the granules produced by the method of vacuum drying, re-dispersed in water to obtain a composition suitable for spraying, including 0,2-2,0% pellets (wt./vol.).

24. The method according to p where water dispersible granular composition intended for biocontrol, is a stable structure that preserves the number of colonies at the level of 108-109CFU/g after a long storage period.

25. The method according to p where water dispersible granular composition intended for biocontrol, is a stable composition comprising a biocontrol agent that retains its phenotype and the ability to actively colonize the grain after a long storage period.

26. The method of reducing the contamination with aflatoxin in food and feed raw materials, VK is uchumi processing said raw materials water-dispersed granular composition, designed for biocontrol, including agent intended for biocontrol, according to claim 1 prepared by a process comprising the stages:
(a) mixing dry ingredients, including sodium carboxymethylcellulose, trehalose and silica clay or mixture of clays until visually homogeneous state;
(b) the suspension of an effective amount of conidia of Aspergillus netoxygen or neoplastigenic, in the peptone solution containing trehalose, for preparation of a solution containing conidia; and
(c) mixing the above dry ingredients with a solution containing conidia, for the preparation of water dispersible granular composition intended for biocontrol,
(d) vacuum drying,
where the sodium carboxymethylcellulose is a binding agent, trehalose is an agent with osmoprotective and adhesive properties, the source of nutrients and a binder agent, silica clay or mixture of clays is a media and nontoxigenic or neoplatonically Aspergillus conidia are an agent, intended for biocontrol.

27. The method of reducing the contamination with aflatoxin in food and feed raw p, where silica clay or mixture of clays includes calcinated kaolin, bentonite, kaolinite or smectite.

28. The way to decrease the possible level of contamination of aflatoxin in food and feed raw materials in item 27, where silica clay or mixture of clays is a soda kaolin clay.

29. The method according to any of PP-28, where the specified raw materials selected from the group including: peanuts, corn, cotton seeds, grain crops, olives and hazelnuts.



 

Same patents:

FIELD: food industry.

SUBSTANCE: invention relates to a food product with extended storage life and to usage of probiotic spores and/or resting cells as a probiotic ingredient in the said product. The product is chosen from the group containing milk products, fruit juices and fruit beverages. The probiotic spores are chosen from the group containing bacterial, yeast and fungous spores. The resting cells are chosen from bacterial, fungous and/or yeast and/or their mixtures. The produced is stored under uncooled conditions during a long period of time by way of probiotic bacteria spores inclusion.

EFFECT: probiotic organisms contained in fruit juices and beverages remain viable during a longer period of time in comparison with live probiotics.

23 cl, 9 ex

FIELD: biotechnologies.

SUBSTANCE: strain of fungus Trichoderma harzianum Rifai VKPM F-180 is used as a producent of an inhibitor of a stimulant of bacterial burn of fruit cultures.

EFFECT: invention makes it possible to reduce losses of decorative and fruit cultures caused by bacteria Erwinia amylovora.

1 ex

FIELD: biotechnology.

SUBSTANCE: fermentation medium for production of recombinant proteins using methanol inducible fungi of species Pichia pastoris is characterised by maintained concentration of urea or its derivatives in the range from about 0.3 M to about 1 M. The medium contains per liter of water basic salts in the following amounts: orthophosphoric acid (85%) from 2.67 to 133.5 ml, calcium sulfate from 0.093 to 4.65 g, potassium sulfate from 1.82 to 91 g, magnesium sulfate-7H2O from 1.49 to 74.5 g, potassium hydroxide from 0.413 to 20.65 g, glycerol from 4 to 200 g, and microelements per liter of water in the following amounts: copper sulfate-5H2O from 0.6 to 30 g, sodium iodide from 0.008 to 0.4 g, manganese sulfate-H2O from 0.3 to 15 g, sodium molybdate-H2O from 0.02 to 1 g, boric acid from 0.002 to 0.1 g, cobalt chloride from 0.05 to 2.5 g, zinc chloride from 2 to 100 g, sulphate divalent iron-7H2O from 6.5 to 325 g, biotin from 0.02 to 1 g, sulfuric acid from 0.5 to 25 ml. The method for production of recombinant proteins comprises the stage of reproduction of methanol inducible fungi of species Pichia pastoris, and the stage of recombinant protein expression using the said fermentation medium in which feeding with methanol is carried out at a rate of about 6 g/l/hr to about 20 g/l/hr.

EFFECT: increased yield of target products.

11 cl, 27 dwg, 16 ex

FIELD: biotechnologies.

SUBSTANCE: preparation with polyfunctional medical-biological activity that influences tissue exchange is produced as a result of liquid-phase depth cultivation of fungus Pleurotus ostreatus All-Russian collection of industrial microorganisms F-819 with further separation of mycelium and cultural liquid and extraction of biologically active substances in the form of a condensed extract from the mycelium. The produced extract, which represents a pharmacological substance of a preparation, is enriched with the agent of the derivative of sterol 4-hydroxy-17P-methylinecisterol with molecular weight of 332.2452 Dalton, polysaccharide β 1-3 glucan and dihydroquercetine with molecular weight of 304.26 Dalton. At the same time the specified components are taken at the following weight ratio: (1):(2):(3):(4) as (79.0-158.0):(1.0-2.0):(10.0-20.0):(10.0-20.0).

EFFECT: invention makes it possible to increase quality of a preparation as a result of increasing its antitumoral activity and reduced toxicity of preparations of antitumoral chemotherapy with simultaneous application with the suggested preparation.

2 cl, 21 dwg, 11 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: entomopathogenic biopreparation for protecting plants from pests contains a dry mixture of a concentrate of a culture fluid based on infection units of entomopathogenic fungi of the type Verticillium lecanii or Beauveria bassiana, or Metarhizium anisopliae, aerosil and an additive which protects the infection units from loss of viability and biological efficiency when drying, with the following ratio of components, wt %: culture fluid concentrate 69.0-77.5, additive - 10.8-13.8, aerosil - 11.6-17.2. The additive is a polyatomic alcohol, carbohydrate, vegetable oil, sodium gluconate and water, with the following ratio of components, wt %: polyatomic alcohol 6.2-13.4, carbohydrate 38.0-41.9, vegetable oil 38.7-41.0, sodium gluconate 0.4-0.5, water 9.3-10.5. The biopreparation is obtained by mixing the culture fluid concentrate with the additive in ratio of 5-7:1, and then adding aerosil in ratio of 4.5-7.6:1 and drying the mixture. Efficiency of the biopreparation on white flies is 95-100% in a working concentration of 1·107 spores/ml; the titre of viable units is (0.35-5.5)·1010 CFU/g.

EFFECT: invention increases biological efficiency of the biopreparation, preserves and increases viability of infection units of fungi during drying and storage.

8 cl, 3 tbl, 15 ex

FIELD: biotechnology.

SUBSTANCE: method of qualitative assessment of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys in operation of spacecrafts and the suspension of spore materials fungi of implementation of the said method is proposed. The test and control samples of aluminium and magnesium alloys are prepared. The prepared samples are dried, sterilised. Fungal cultures - strains of microorganisms Paecilomyces variotii Bainier of All-Russian collection of microorganisms F-4039D, Ulocladium botrytis Preuss of All-Russian collection of microorganisms F-4032D, Penicillium chrysogenum Thorn of All-Russian collection of microorganisms F-4034D, Aspergillus sydowii (Bainier et Sartory) Thorn et Church of All-Russian collection of microorganisms F-4037D, Cladosporium sphaerospermum Penz. of All-Russian collection of microorganisms F-4041D are inoculated for growing spores into tubes with a sloped agar medium of Czapek-Dox. The tubes are thermostated at a temperature of (29+2)°C for 14-28 days until appearance of mature spores. Spore materials suspensions of individual cultures of the said fungi are prepared, which are then mixed in equal proportions. And the concentration of spores of each fungi species in the suspension should be in the range of 1-2 mln/cm3. The resulting suspension is applied to the sterilised test samples. Then they are dried and placed as one sample in each Petri dish on the surface of agar medium of Czapek-Dox. The control samples not treated by the spore material are also placed on the surface of the agar medium of Czapek-Dox in the Petri dishes. The Petri dishes with the test and control samples are placed in different desiccators at the bottom of which water is poured to maintain the humidity more than 90%. The desiccators are closed and incubated in the thermostat at a temperature of (29±2)°C. The exposition of test and control samples is carried out for 40, 120 and 180 days. Then the samples are removed from the Petri dishes, and the corrosion products and mycelium are removed from the surface of the samples washing them in running water. The samples are soaked in 70% ethyl alcohol for 30 minutes, then they are washed with detergent and dried. Then, using a scanning electron microscope a qualitative assessment of biocorrosion damages is carried out. According to the change in the appearance of the surface of the samples the initial stages of biocorrosion and its type is evaluated, as well as the distribution of corrosion damage on the surface of the samples, and the dependence of the biocorrosion process on time is determined.

EFFECT: inventions enable to carry out the qualitative assessment of the initial stages of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys with the thickness of more than 2 mm.

2 cl, 9 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to microbological industry, particularly production of biologically active substances and can be used in producing fusicoccins for different purposes, for both agricultural and medical purposes. The culturing medium contains molasses, raw sugar, potassium nitrate (KNO3), potassium phosphate (KH2PO4), magnesium sulphate (MgSO4 x 7H2O) and tap water in a given ratio.

EFFECT: invention increases the range of fusicoccins.

4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a compound and a pharmaceutically acceptable salt thereof to be used as an antifungal agent, particularly, a therapeutic agent for deep fungal disease. The fungus Acremonium persicinum is collected, and a cyclic compound is recovered from its cultural fluid.

EFFECT: what is presented is the compound applicable as an antifungal agent.

10 cl, 16 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacology and medicine, and represents a method for preparing a concentrated culture fluid of active anamorphic fungi species P. farinosus, S. militare, V. brongniartii with antibiotic activity, including growth of the fungi Cordyceps in liquid nutrient media (Czapek's medium, cereal infusion, unhopped wort (7%)) in the steady-state environment followed by concentrating of the culture fluid, characterised be the fact that test insects are further infected intra-hemocoel by G. mellonella (introduction of the suspension 0.01 mL into a hemocoel), immersion of larvae into the suspension for 30 seconds, infection of feed with the fungi Cordyceps for preparing the anamorphic virulent forms; and then biotechnological cultures the concentrated culture medium of which has high antibiotic activity is prepared from the mummified and melanised larvae G. mellonella by fungi re-inoculation from the insects on the liquid nutrient media.

EFFECT: invention provides unifying the process of the concentrated culture fluid of the anamorphic fungi Cordyceps.

3 ex, 3 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry and use of a Trichoderma harzianum Rifai strain as a producer of an Impatiens necrotic spot tospovirus inhibitor. The Trichoderma harzianum Rifai strain, which is deposited in the Russian National Collection of Industrial Microorganisms under No.F-180, is a producer of the homogeneous enzyme L-lysine-a-oxidase, which exhibits marked antiviral activity with respect to Impatiens necrotic spot tospovirus.

EFFECT: reduced loss of decorative and vegetable crops caused by Impatiens necrotic spot tospovirus.

2 ex

FIELD: biotechnologies.

SUBSTANCE: strain of fungus Trichoderma harzianum Rifai VKPM F-180 is used as a producent of an inhibitor of a stimulant of bacterial burn of fruit cultures.

EFFECT: invention makes it possible to reduce losses of decorative and fruit cultures caused by bacteria Erwinia amylovora.

1 ex

FIELD: chemistry.

SUBSTANCE: entomopathogenic biopreparation for protecting plants from pests contains a dry mixture of a concentrate of a culture fluid based on infection units of entomopathogenic fungi of the type Verticillium lecanii or Beauveria bassiana, or Metarhizium anisopliae, aerosil and an additive which protects the infection units from loss of viability and biological efficiency when drying, with the following ratio of components, wt %: culture fluid concentrate 69.0-77.5, additive - 10.8-13.8, aerosil - 11.6-17.2. The additive is a polyatomic alcohol, carbohydrate, vegetable oil, sodium gluconate and water, with the following ratio of components, wt %: polyatomic alcohol 6.2-13.4, carbohydrate 38.0-41.9, vegetable oil 38.7-41.0, sodium gluconate 0.4-0.5, water 9.3-10.5. The biopreparation is obtained by mixing the culture fluid concentrate with the additive in ratio of 5-7:1, and then adding aerosil in ratio of 4.5-7.6:1 and drying the mixture. Efficiency of the biopreparation on white flies is 95-100% in a working concentration of 1·107 spores/ml; the titre of viable units is (0.35-5.5)·1010 CFU/g.

EFFECT: invention increases biological efficiency of the biopreparation, preserves and increases viability of infection units of fungi during drying and storage.

8 cl, 3 tbl, 15 ex

FIELD: chemistry.

SUBSTANCE: invention relates to plant protection agents. Disclosed is a preparation having fungicidal properties, where the active components used are terbuconazole, imazalil and prochloraz or metalaxyl. The ratio terbuconazole:imazalil:prochloraz/metalaxyl is equal to 1:(0.5-15.0):(0.5-15.0), with consumption rate of active substances of 45-300 g/t seeds.

EFFECT: invention widens the range of biological action against snow mould, increases biological efficiency against root rot.

4 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry and use of a Trichoderma harzianum Rifai strain as a producer of an Impatiens necrotic spot tospovirus inhibitor. The Trichoderma harzianum Rifai strain, which is deposited in the Russian National Collection of Industrial Microorganisms under No.F-180, is a producer of the homogeneous enzyme L-lysine-a-oxidase, which exhibits marked antiviral activity with respect to Impatiens necrotic spot tospovirus.

EFFECT: reduced loss of decorative and vegetable crops caused by Impatiens necrotic spot tospovirus.

2 ex

FIELD: biotechnologies.

SUBSTANCE: strain Trichoderma harzianum Rifai, having L-lysine-alpha-oxidase activity is deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under the registration number RNCIM F-180 and may be used in agricultural biotechnology and plant growing.

EFFECT: invention makes it possible to reduce losses of decorative and vegetable crops.

2 ex

FIELD: agriculture.

SUBSTANCE: agent comprises salicylic acid, lipids from mycelium of fungus Ascichyta pinodes and the source of magnesium - magnesium sulfate MgSO4 or magnesium chloride MgCl2 at the following ratio of components: wt %: salicylic acid - 0.001, lipids from mycelium of fungus Ascichyta pinodes - 0.0001; the source of magnesium (magnesium sulfate MgSO4; or magnesium chloride MgCl2) - 0.001; water - 99.9979.

EFFECT: invention enables to increase the growth indices of pea, yield, disease resistance.

7 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry, particularly protection of plants from diseases caused by phytopathogenic bacteria and phytoplasma. The disclosed agent is obtained by preparing a producer inoculum, growing the producer on a fermentative medium, drying the culture fluid, extracting a macrolide complex using lower alcohols, concentrating the extract and mixing said extract with additives and solvents in the following ratio, wt %: macrolide complex 17.0-23.5, solvents and additives - up to 100, where the producer used is a Streptomyces fradiae ARRIAM-53 strain.

EFFECT: invention increases plant protection effectiveness.

2 tbl, 10 ex

FIELD: chemistry.

SUBSTANCE: invention relates to bioengineering and specifically to a method of producing an entomopathogenic preparation. The method involves submerged cultivation of Verticillium lecanii fungus on a culture medium, followed by concentration of the culture fluid and adding a polyfunctional additive containing the following in wt %: potato starch 6-8, titanium dioxide 2-5, boric acid 0.01-0.03, streptomycin 0.004-0.008, soya lecithin 8-14 or liquid petrolatum 4-6, water - the rest.

EFFECT: invention increases biological effectiveness of the bio-preparation and preserves activity of the bio-preparation during storage for 4 months.

1 tbl

FIELD: agriculture.

SUBSTANCE: remedy for plant protection against bacterial and fungus diseases contains a complex of streptotricin compounds formed by Str. griseus or Str. Lavendulae geophilous fungus as a bactericide agent with the fungicide agent represented by carbendazime as well as solvents and functional additives. It is preferable that one use xanthane gum, sulphanole and benzyl alcohol as a thickener, a disperser and a preservative accordingly.

EFFECT: improvement of properties of the remedy for protection of plants.

2 cl, 4 tbl, 10 ex

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to biotechnology and can be used as plant protection agents. The method involves selecting and growing Arthrobotrys conoides Dreschsler fungi in a suitable culture medium. A jellifying agent and at least one carrier are added to the culture medium to obtain a mixture. The obtained mixture is jellified through reaction with a calcium salt containing solution which is added in drops. The obtained granules are dried to moisture of 13-18%.

EFFECT: method enables to obtain granules which ensures stability and survival of mycelial fungi.

14 cl, 3 ex

FIELD: biotechnology.

SUBSTANCE: the present innovation deals with immunization in plants, particularly, compositions and methods for inducing plant resistance to phytopathogenic organisms, such as phytopathogenic fungi. An agent indicated for inducing plant resistance to phytopathogenic microorganisms is being an extract out of biomass of non-phytopathogenic microorganisms. The method to obtain the above-mentioned agent includes the following stages: a) resuspending against 50-200 g (dry weight) biomass of non-phytopathogenic microorganisms in 1 l either inorganic or organic solvent, b) mixing at room temperature for 1-12 h, c) incubating, d) resuspending, e) cooling up to room temperature at maturing and f) filtrating, not obligatory. The innovation enables to induce plant resistance to phytopathogenic microorganisms.

EFFECT: higher efficiency of immunization.

12 cl, 6 ex, 5 tbl

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