RU2074158C1 - Strain of bacterium azotobacter chroococcum used for preparing bacterial fertilizer for amatanth - Google Patents

Abstract

FIELD: agriculture microbiology, bacterial fertilizers. SUBSTANCE: invention proposes the strain of bacterium Azotobacter chroococcum that can be used for preparing the bacterial fertilizer for amaranth. Use of the proposed strain increases culture productivity by 70-75%. EFFECT: increased productivity of culture. 2 tbl

Description

 The invention relates to agricultural microbiology and is a new strain of nitrogen fixer Azotobacter chroococcum, which can be implemented in the microbiological industry to produce various biological fertilizers.

 A known strain of Azotobacter chroococcum T-12 (deposited in the Central Museum of industrial microorganisms of the All-Russian Research Institute of Genetics for N B-3344), designed to obtain bacterial fertilizers. The disadvantage of this known strain is the specificity of its action, which extends mainly to nightshade (in particular tomatoes) and the absence of any positive effect in terms of increasing the yield of amaranth crops, as well as the relatively high nitrate content in plants grown with its use. In addition, bacterial fertilizers made on the basis of this strain must necessarily contain peat filler.

 Also known is the Azotobacter chroococcum strain deposited at the Central Museum of Industrial Microorganisms of the All-Russian Research Institute of Genetics for N В-3173, intended for the production of bacterial fertilizer4. This strain in terms of the combination of morphological and physiological and biochemical characteristics, as well as in terms of the achieved technical effect, is the closest to the object we are declaring and therefore we have chosen as a prototype. The disadvantages of this known strain are the specificity of its action, which applies only to barley and cereals and cereals close to it and the lack of a positive effect in increasing the yield of amaranth (in particular, amaranth crimson), as well as the relatively high nitrate content in all parts grown with its use plants. In addition, this strain multiplies and functions in the rhizosphere of amaranth for no more than 1.5 months, which reduces its effectiveness in terms of increasing productivity.

 The aim of the present invention is to increase the yield of amaranth, reducing the nitrate content in the resulting green mass and increasing the life of the strain in the atmosphere of amaranth.

 The declared goal is achieved through the use of a new strain - Azotobacter chroococcum Ш 25, isolated from the resosphere of crimson amaranth (Amaranthus crueutus) when growing on sod-podzolic soil with pH 5.5 6.0. The new strain was deposited at the Central Museum of Industrial Microorganisms of the All-Russian Research Institute of Genetics under N В-6011.

 The inventive strain Azotobacter chroococcum W 25 has the following individual characteristics.

 Morphological signs. The cells of the strain at a young age are small cocci and diplococci; depending on their age and growth environment, their size ranges from 0.7 to 1.7 μ. Cells can be oriented singly, in pairs or short chains, the tendency to the formation of which increases with age. Weakly mobile, the capsule around the cells is invisible. On solid media forms typical colonies. The resting form of the cyst. With aging, the culture of the strain acquires a brown color, which appears at the age of 2 3 days. The pigment does not penetrate the medium; cells can be stained with various Gram aniline dyes.

 With the growth of the strain Azotobacter chroococcum W 25 in Burke culture medium, the stroke of a 2-day-old ivory culture is opaque with a shiny surface; rounded colonies with smooth edges, opaque, also ivory.

Physiological and biochemical characteristics. Strain Azotobacter chroococcum Ш25 - aerob, optimal growth temperature 28 30 ^o C; minimum 15, maximum 45 ^o C. Reaction of the strain to pH: optimum 6.9 7.5, minimum 5.8, maximum 8.8. Strain B-6011 assimilates the following carbohydrates: glucose, levulose, dextrin, maltose, sucrose, arabinose, xylose, mannose, galactose and alcohols: mannitol, ethanol, glycerin. Does not assimilate: salts of formic, salicylic, oleic acid, methanol, phenol, cellulose, lignin. It uses free, ammonium, nitrite and nitrate nitrogen for its development, and the latter is first transformed by the strain into ammonia, and then it is used by an azotobacter. On the IPA is growing well.

Synthesis products on Burka culture medium after 4 days of incubation at 28-30 ^o C: B vitamins (biotin 0.09, pyridoxine 1.5, thiamine 0.9 mg / l), indolylacetic acid 69.0 mg / l; nitrogen fixation of 0.45 μg nitrogen / ml per 1 h.

 The strain Azotobacter chroococcum W 25 has the ability to attach to the root surface of various amaranth (Amaranthaceae), in particular amaranth crimson (Amaranthus cruentus), where it multiplies and functions for 4-6 months after inoculation.

 Previously, the strain of the microorganism Azotobacter chroococcum W 25, registered at the Central Museum of Industrial Microorganisms of the All-Russian Research Institute of Genetics for N В-6011, with the above characteristics was not described in the literature, therefore, we can confirm the compliance of the claimed object with the first criterion feature of the invention of "novelty". And finally, our proposed microorganism strain is easily cultivated, its production without special difficulties can be established on an industrial scale and, therefore, we can talk about the presence of the third criterion of the invention “industrial applicability” in the claimed object.

 The strain of Azotobacter chroococcum W 25 that we declare as a bacterial fertilizer can be demonstrated in the following examples.

 EXAMPLE 1

 Weekly seedlings of crimson amaranth (Amaranthus cruentus) growing in a liquid nutrient medium are treated with a suspension of Azotobacter chroococcum W 25 cells and then grown in a room greenhouse for 1 day. Then the roots are separated and sequentially washed 7 times in Burke's medium; Further, weighed portions of the roots are ground, diluted and plated on the above nutrient medium. Data on the number of cells of the strain in 1 g of roots for this case are presented in table 1.

 EXAMPLE 2

 Perform as example 1, but the seedlings of Amaranthus cruentus are treated with a suspension of cells of the strain Azotobacter chroococcum N B-3173.

 EXAMPLE 3

 Carried out according to the scheme of example 1, but instead of the strain indicated therein, Azotobacter chroococcum T-12 is used.

 Of the presented in table. 1 data it is clearly seen that the number of cells of the claimed strain in 1 g of the roots of Amaranthus cruentus is 8 10 times more than the strain of the prototype, and 30 50 times more than the strain of the analogue, which clearly indicates a much better ability of the strain N B-6011 is attached to the surface of the roots of amaranth crimson. Note that similar results were obtained by us on other amaranth species (Amaranthus candatus, Amaranthus mantegazzianus).

 EXAMPLE 4

A strain of the microorganism Azotobacter chroococcum W 25 is grown in a fermenter in modified Burke medium to a cell concentration of 10 ⁸ 10 ⁹ cells / ml. Then, Amaranthus cruentus seeds are bacterized at the rate of 1 liter of preparation per portion and seeds that are optimal for sowing 1 ha of land, after which they are sown in soil traditional for the non-chernozem zone of Russia with a pH of 5.8 6.5. The duration of the growing season is 4.5 months. At the end, it is harvested in the form of green mass and its parameters such as the average total plant weight, its height, total yield and nitrate nitrogen content in various parts of the plants are determined. All these data are presented in table. 2.

 EXAMPLE 5

 Perform as example 4, but using a strain of Azotobacter chroococcum B-3173.

 EXAMPLE 6

 Carried out according to the technological scheme of example 4, but using a strain of Azotobacter chroococcum T-12.

 EXAMPLE 7 (comparative).

 A culture of Amaranthus cruentus is grown according to the scheme described in example 4, but without bacterialization of its seeds by any strain.

 As can be clearly seen from the data given in Table 2, the use of the strain Azotobacter chroococcum Ш25 we declare allows us to significantly (70 75%) increase the yield of Amaranthus cruentus, as well as reduce the nitrate content in various parts of plants (which is especially pronounced in the case of stem stalks by weight most of the green mass obtained during harvesting). In addition, the life time of the strain we declare (and, consequently, the duration of exposure to bacterial fertilizer) is no less than the duration of the growing stage of Amaranthus cruentus (> 5 months), while in prototype strains it is only 1.2 and 1.5 months, respectively. Note that we obtained similar results in the case of other representatives of the Amarantha ceae family (Amaranthus candatus, Amaranthus mantegazzianus), so that the strain we declare can be used as a bacterial fertilizer for a wide variety of amaranth cultures.

Claims (1)

 The bacterial strain Azotobacter chroococcum VKPM B-6011 used to obtain bacterial fertilizer under amaranth.

Images