UA73682C2 - Insectofungicide biopreparation for pest and disease control of farm crops - Google Patents

Abstract

The invention relates to biotechnology, namely to obtaining of complex biopreparation for pest and pathogen control of plants. Ecologically safe complex insectofungicide preparation is produced on the base of such two strains as Pseudomonas aureofaciens IMB B-7096 and Pseudomonas aureofaciens IMB B-7097. Both strains are characterized by entomopathogenic, antifungal and antibacterial activity resulting from synthesis of pigments of phenazine series and by stimulation effect on plants growth. The preparation can be used for control of vine fungus diseases, root rots of other farm crops (wheat, barley, pea), as well as for treatment of harvest of vegetable crops while storing.

Description

Description of the invention

The invention relates to biotechnology, namely to the preparation of a complex biological preparation for the protection of plants from pests and pathogens.

Biological means of plant protection, which occupy an increasingly significant place in the market of biotechnology products, are usually monovalent and are directed only against insect pests or microorganisms - plant pathogens. At the same time, the creation of complex biological preparations based on microorganisms, which have several types of useful activity, is quite attractive. Microorganisms that are promising from this point of view include strains of 70 Rvetsdotopav ashgeogasiepv, the antifungal properties of which are devoted to many reports | Smirnov,

Kyprianova, 1990; VeskKeg ei aiI., 1990, Tpotazpom/ ei aiI., 1990). We have shown that, along with high antibacterial and antifungal activity, some strains of this species have pronounced entomopathogenic properties.

The task of the invention is to create a biological drug with 79 complex antimicrobial, antifungal, entomopathogenic and growth-stimulating effects for the protection of agricultural crops from diseases and pests based on active strains of Rzeidotopaz aigeoiasiepz.

The drug Agat 25K for increasing plant yield (Patent of the Russian Federation Mo2111196 dated 05.20.98) can be considered as a prototype of the proposed drug, which contains inactivated bacteria Rzeiidotopaz ashygeoiasiepz strain H1I6 in the form of a paste. The drug is used for seed treatment and spraying of barley, wheat, and sugar beet crops. It inhibits the development of root rot, septoriosis, fusarium, brown rust, thus increasing the yield of agricultural crops. However, this drug contains only one strain of bacteria with a limited spectrum of antifungal activity directed against several types of fungal pathogens.

As an analogue of the drug, the bacterial strain Rzeidotopaz ashtsygeoiasiape 51 can be considered (Patent of the Russian

Federation Mo2203945 dated 10.05.2001), used for pre-sowing treatment of wheat seeds. The drug, c 22 obtained on the basis of a strain isolated from arable soils of Bashkortostan, inhibits the development of wheat diseases caused by hard soot and root rot. However, this is a monovalent drug consisting of one strain of bacteria, it has no entomopathogenic activity and does not affect harmful insects, and the pigment with which the antimicrobial and growth-stimulating effect of the drug is associated has a fairly moderate antimicrobial activity. Therefore, the spectrum of action of the drug is limited, and its use gives positive results only in wheat diseases caused by fungal pathogens. with

The invention was based on the task of creating an ecologically safe complex insecticidal preparation based on Rzepdotopaz atsgeoiasiep5z strains, which would ensure its wide application, use for the protection of vegetable, cereal, fruit crops, grape plantations, forest areas from insect pests and fungal and bacterial diseases and stimulation of plant development. 3o The task is solved by the fact that the biopreparation contains cell biomass mixed with the medium for growing two strains: Rzeidotopaz ashygeoiasiep5 UKM B-111 and Rzeidotopaz ashygeoiasiepz UKM B-306.

The named strains were selected as a result of a large-scale study of a significant collection of microorganisms, in particular representatives of the Rzeidotopaz ashgeoiasiepv species, for antagonistic activity against phytopathogenic fungi and bacteria. They were characterized by significant antibiotic activity due to the synthesis of antibiotics - C derivatives of phenazine (Table 1). c Entomicidal activity of different strains of Rzepdotopaz ashgeoiasiepz against the caterpillar of the 5th instar apple borer

Iz" fluctuated widely, reaching 90905 in individual strains and in different experiments (Table 2).

As can be seen from the data presented in the tables, strain B-111 showed the most significant antifungal properties, and strain B-306 caused the highest percentage of death of apple borer and Colorado potato beetle. Significant entomocidal activity of the named strains against a number of pests was revealed, and to a greater extent it was manifested in the B-306 strain. (Table 3). None of the strains was antagonistic to the other; they grew well in a mixed culture. On the basis of both these cultures, a complex drug called haupsin was created. b Both strains are deposited in the Depository of the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine under numbers 1MV bo 20 V-7096 and 1MV V-7097.

The method of obtaining haupsin includes the joint cultivation of both strains in deep culture on an organo-mineral medium with a content of total nitrogen 1.8-2.595 (including amino nitrogen 0.15-0.3090), reducing substances 1.8-2.595, compounds phosphorus 0.04-0.0795, potassium 0.05-0.0795, magnesium 0.005-0.0195. The source of organic compounds is the by-products of the food industry. Broth 59 cultures of strains B-111 and B-306, obtained by growing on a rolling pin, with a titer of 5x109 - 1x1079 cells/ml are used as seed material.

GF) Seed material in the amount of 595 from the total volume of the medium was introduced into a 100-liter fermenter with 7 7Ol of the medium of the above composition. Cultivation temperature - 25-27 2, aeration - 1 volume of air per 106. medium / min., stirring - 250 rpm, duration of fermentation - 24 hours.

The joint growth of bacteria in conditions of aeration is accompanied by intensive synthesis of antibiotically active phenazine pigments, which is evidenced by the orange-red color of the culture liquid.

The preparation obtained in the fermenter contains 1.0-1.2x10!! cells per ml and is not inferior in productivity to preparations obtained by separate cultivation of strains B-311 and B-306 (Table 4)

Haupsin is non-virulent and non-toxic for laboratory animals and humans, does not have a negative effect on beneficial organisms of the biocenosis, can be used in any phase of plant development (flowering, harvesting).

The liquid form of haupsin is a native culture liquid with cells, obtained by growing in cylinders on a rocker or a culture liquid with cells from a fermenter, diluted with an aqueous solution of a stabilizer to a titer of 1x1079 cells/ml, characterized by a high level of activity during three months of storage at room temperature.

With the use of rifampicin-resistant mutants of Rzeidotopaz ashygeoiasiepz, it was established that when spraying the leaves of plants, haupsin remained in high titers for 7 days.

The results of field tests of the liquid form of haupsin are shown in Table 5.

The drug at the consumption rate of 2 zlotys/ha effectively reduced fruit damage by apple fruit borer (92-9390) and fungal diseases (94-9695). In general, haupsin was not inferior to chemical pesticides in terms of effectiveness in relation to fruit-eaters, inhibition of 70 fungal diseases, yield and profitability. It was established that, along with its insecticidal and fungicidal properties, haupsin had a stimulating effect on plants (table. b).

Pre-sowing treatment of seeds with haupsin not only reduced the damage of plants by root rots, but also increased their resistance to powdery mildew on cereals, late blight on potatoes, late blight and macrosporiosis on tomatoes during the growing season, which contributed to increasing the yield of these crops by 14-70905.

In recent years, haupsin has also proven itself against such diseases and pests as oidium, mildew, black spot, gray rot, powdery mildew, ascochitosis, verticillosis, septoriosis, fusarium, peronosporosis, hard soot, many types of leaf curlers, aphids, mealybugs, bonfire, fruit eater, etc. Haupsin not only protects plants, inhibits the development and spread of the disease, but also has a positive effect on the plant itself, "treats" it.

Haupsin is successfully used for pre-sowing treatment of seeds, soaking seedlings, treatment of vineyards and forest areas from pests.

It has been established that strains of Rzeidotopaz ashgeoiasiepz are resistant to chemical plant protection agents, therefore haupsin is compatible with most fungicides and can be used in the form of tank mixtures. with strains of Rzeidotopav atsshgeoiasiapie

The activity and the substances that determine it are sch 5 pet nn introduced acid

Zoyufenyain 10000106 20700006 000

Ttsroesyut uyent 00000530 105300 40130802 la v z Mepoyutavnyae 21680) 00160806 00700

Nevtiovrotit stelchyaest 80530530 53005530 0530 53030 la

Mshsorryutsht 00000700190190 30060050 from 53013 in

Antagonism against phytopathogenic bacteria" (Adgobasivtimt isteasiepo 060201 »30 530) 5 714 | 23020 | »30 o | «

Etmiasayuyuyuv 00000000 00000 000 t

Etpaayuueya 00000012 10 2207 2088. - vmeste tispoyanetiav 00800530 530 530010 5300530 5300 190530 th» Hatotopev chelstuta 00505300 530 30005308 16053000

Resolved solution 1711 11111111 15 on rvv|lv vvovi - vva 00111112 402 018 ml o cut 00000000 40902 80790822 o vesutae 00001000 LE " zone of growth retardation of test culture in mm trace; (Fe) 20 "x substance is present in chromatograms

Ko) caterpillars of the apple borer and larvae of the Colorado potato beetle

M strain of Apple caterpillar Larvae of the Colorado potato beetle, mortality, 95 | imago, 96 mortality, 95 | imago, 90 in 91 bo va 00010ю00ю1ю vv 11ю1ю1ю v 65 v-114 vh 38 BO" BO

Control (without processing) 25 75 25 74 nsrove 1111 " Data, the reliability of which compared to the control is proven at the 9595th level "x Data, the reliability of which compared to the control is proven at the 9995th level

Table C

Entomicidal activity of Rzendotopav ashgeoiasiepv strains

Type of insects Stage of development SCvo" for MoMo strains 19 v-306 v-111

Chichasta lityuuta 011101201о6 i ruvovavotya 00000100014000000060 7 cm

I

"SCvo - the number of cells in million/ml, which causes the 5095th death of insects

With co

Productivity and biological activity of Rzepdotopav ashgeoiasiepv strains

UKM B-111 and UKM B8-306 with separate and simultaneous I«o) cultivation under aeration conditions

M strain |Productivity (billion cells per ml) after 24 hours. of growth on Diameter of the zone of growth retardation |Mortality of apple caterpillars on corn-molasses environment Ryvzagiit ochuvrogyt fruit-eater with : : in. in cylinders on in the fermenter with correction for mortality in the rocking chair control in-zoe tz2:odo patio? both: « v-111-8-306 12.9--1.11 119.0-- 7.8 91,245.7 -v . 2 I

The use of haupsin to protect the orchard from apple fruit borer, scab and fruit rot

Year | Variant | Rate of expenditure Multiplicity | Yield, Affection of the fruits of the harvest crop, 9 pollinator | Biological - 1 l/ha of treatments t/ha - disease efficiency 95 oz tab Kontol! 0000101863000000000630000100002790001g

F o m r e m tauto 09000108 01000006Y00000000001000009160000000000100100000 m tt tags 01019 Pages "One full generation of apple fruit" X two generations expenditure norms: Zolon, Karate, Sherpa, Polycarbatsin, Skor

Table 6

The protective and growth-stimulating effect of haupsin on agricultural crops 2 Culture | Method of application Norm Year Similarity Reduction of damage to plants, 96 PIncreasing the yield of seed consumption, 95 to

Claims (1)

The formula of the invention is part 6) Insectofungicidal biopreparation for combating pests and diseases of agricultural crops, which differs in that it contains strains of Rzepdotopaz atsgeoiasiepz IMV B-7096 and Rzeidotopaz atsgeoiasiepz IMV B-7097. c Official Bulletin "Industrial Property". Book 1 "Inventions, useful models, topographies of integrated circuit microcircuits", 2005, M 8, 15.08.2005. State Department of Intellectual Property of the Ministry of Education and Science of Ukraine. (Se) (ze) and - - and? -i (95) (o) (ee) Ko) ime) 60 b5