SU686707A1 - Bacillus thuringiensis var galleriae h5bt-103 strain - Google Patents

Description

(54) BACILLUS THURINGIENSIS VAR. Galleriae

H5BT-103

Claims (2)

This invention relates to agricultural microbiology and is a new strain of you. Ihuringiens Js used to produce entobacterin used to kill insect pests. It is known that one of the means for the destruction of insect pests is the bacterial preparation ENTO bacterin, the production of which occurs on the help of the strains of Bac. tHuringieMsis Ulp galleriae 1. At the present time, microbiological industry uses enteroconstrictor bacterium to produce anthoacterin. The lhurin9ieTisis of the 5th, isolated from the dead insect pest larvae 2. However, these are wild phage producing you, thuringieTisis, do not withstand large amounts of production enzymenters and are rejected due to lysis during the first commission checks. The aim of the invention is the strain You. thuringfensis var. galleriae H5WT-103, answerI1-1y all necessary. requirements imposed on manufacturing shp11mm-producers of entobacterin, i.e. under deep cultivation conditions, it does not produce virulent phage and allows to obtain products of higher quality. The resulting strain you. thuringiensis ag. galleriae H5WT-103 has the following morphological and physiological characteristics. Morphological signs. The size of the cells in the exponential phase of development on ordinary SPA type agar is 4.0-6.0 x 2.0-2.5 µm. The predominant form is rod-shaped. Under conditions of submerged cultivation on liquid nutrient media, cells are slow-moving, occur in the form of short and long chains. Peritrichs are gram positive. During the transition from the exponential phase to the stationary phase, they form oval cnofoi 1.8-2.2 microns in size and protein, in the form of bulbs, ethocidal crystals measuring 2.2 x 2.5 microns. According to the H-antigen, the H5BT-103 strain belongs to the 5th serotype, 5a-5b. Physiological signs. Organic medium for supporting selection and reproduction is dry nutrient agar type SPA. Colonies in & forme, convex with rhizoid edges, The diameter of the colonies of the cult of the culture reaches O, 6, O9 cm. The pigment does not form. Nutritious broth. Everywhere is significant, the precipitate is filamentary or veil on the fourth or sixth day. Litmus milk. Peptonization and alkalization from the top (24-48 h), then acidification and reduction of milk, coagulation on the third to fourth day digestion is weak on the sixth day. Potato slices. The culture is dry yellow. Agar with gelatin. The hydrolysis zone is 2-3 mm. Agar - casein .. Enlightenment partial. Agar-starch. Idrolysis zone 5-6 mm. . Yeast polysaccharide regulated nutrient medium used for the production of entomopathogenic bacterial preparations. Intensive growth is observed by 12-14 hours of cultivation at 28-30 C. Spore formation and precipitation is completed by 36-42 hours of growth. The titer of viable spore in the culture fluid at the end of cultivation on depleted DPS (2% EBK-J-1% flour) reaches 2.5-3.0 billion / ml. Relation to carbon sources. Absorbs glucose, cellobiose, maltose, glycerium and salads without releasing gas. Does not digest sucrose / mannose, arabinose, inulin mannitol, sorbitol, dulcite, acetates, citrates. Relation to phages. Own phages do not fall. Resistant to viral phage type FD-2 Pi-13. Shows antagonism to you, thuringiensis to 4,1,3-serotypes. - to obtain the strain H5BT-ld3. used the principle of gene recombination, i.e. redistribution of determinants of the genome of cells in the range of cellular salinization of the parent population. As a factor inducing this redistribution, we used the temperature difference minus 10 plus. As an initial culture, from which the H5BT-103 strain was obtained, a conversion variant of the 1st serotype HI BT was found. storage in VNIIbakpreparat. The table presents the data on the productivity of the H5BT-103 mtam in comparison with the data on the productivity of strains 69/6 prototype and the HIBT parent strain. As can be seen from the table, the head H5BT-103 differs from the parent in relation to the serotype and is therefore the producer of another e-pathogenic drug. In addition, the H5BT-103 in comparison with the strain of the rototype produces twice as much crystalline complex, which will significantly reduce the consumption of raw materials for the production of entobacterin and in connection with this and reduce its cost. Tamm H5BT-103 is a phage-producing culture, and it exceeds: Tamm prototype is virulent and more than 6 times. Claims of the invention: Strain Bacillus thurinqiejisis var. ST0.1 ler-iaeHSBT-lOB, is used for the manufacture of the entomopathogenic drug entobacteric. Obtained from you. ihutingiensis of the first serotype H1BT-2 by recombination of genes when exposed to temperature minus plus. Stored in the collection VNIIbakpreparat. Morphological signs. The size of the cells in the exponential phase of development, on a conventional agar of the SPA type, is 4.0-6.0 x 2, 5 µm. The predominant form is rod-shaped. Under conditions of submerged cultivation on liquid nutrient media, the cells are not mobile. They are found in the form of short and long chains. Peritrichia are gram-positive. During the transition from the exponential phase to the stationary phase, ovalBL spores of 1.82.2 μm in size and protein-like entsmotsidny crystals of 2.2-2.5 μm in size form. According to the H-antigen, the H5BT-103 strain is referred to the 5th serotype, 5a-5b. Physiological features, brethan- "environment for dl. supportive selection and reproduction - dry nutrient agar of the SPA type. Colonies in the R-shape are convex with p isoidyg “1 edges. 1Diameter of COLONY 5-day culture reaches 0.6-0.9 cm. The pigment does not form. Nutritious broth. Cloudiness significant, sediment filiform or on the fourth or sixth day. Litmus milk. Peptonation, understroking from above 24-48 hours, then subcrescence and reduction of milk, coagulation on the third or fourth day, digestion weak on the sixth or eighth day. Potato slices. The culture is dry, yellow. Agar with gelatin. The hydrolysis zone is 3-2 mm. Agar casein. Partial enlightenment. Agar-starch. The hydrolysis zone is 5-6 mm. Yeast polysaccharide is a regulated, reliable and environment used for production of pathogenic bacterial preparations. Intensive growth is observed for 12-14 hours of cultivation at 28-30 ° C. Sporulation and rash complete by 6-42 hours of growth. The titer of viable ctiof in the culture fluid at the end of cultivation on depleted DPS (2% BVK + 1% flour) reaches 2, .5-3.0 billion / ml. Relation to carbon sources. Absorbs glucose-free gas Comparative characteristic productivity of the H5BT-103 strain and the H1BT-2 strains parent and 69/6 SPROTOTIP) H5WT-103 U 2.5-3.0 Bulb He H1ST-21 3, 0-4.0Romb parent 69 / 6U 1.2-1, 8 Bulb Pg-l Prototype Sources of information taken into account in the examination 1, Aleshina O.A. and others. The biological method of combating pests and diseases of agricultural plants. .Knies 1975, p. 33-37, cellobioe, maltose, glycerin, bksalata. Does not absorb sucrose, mannose arabinose lactose, inulin, mannitol, sorbitol, dulcite, acetates, citrates. Attitude to phages1m. Own phages do not produce. Resistant to virulent phages like FD-2 and Fi-l3. Antagonism to you. thuriwgicnsjs 1,3 and 4- serotypes. Not 2. M. Choverda and Simonov L.A. The selection of different variants of the bacilli of the Turin yenses group from one the same insect. The use of microorganisms in animal husbandry and for plant protection, l., 1968, p.123-126. Resistant 0.08 produces products Sustainable about O to Fg-lL rips L Sensitive - 0.5 flax