RU2413762C2 - STRAIN OF RHIZOBIA Bradyrhizobium japonicum BKM B-2456D FOR MANUFACTURING BACTERIAL FERTILISER FOR SOYA - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: strain of rhizobia Bradyrhizobium japonicum BM-85k is isolated from natural population of rhizobia, spread in soils of southern zone of Amur region and is deposited in All-Russian Collection of Microorganisms of G.K.Skryabin Institute of Biochemistry and Physiology of microorganisms (IBPM) of Russian Academy of Sciences under registration number "ВКМ В"-2456D.

EFFECT: increase of soya beans yield and improvement of its quality.

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Description

The invention relates to agricultural microbiology and can be used for the production of bacterial fertilizers for soybeans.

Closest to the invention is a strain of Bradyrhizobium japonicum 648A used for the preparation of soya nitragin (AS USSR No. 1524432).

Known strains have a relatively low level of virulence and activity and only mannitol is used as a carbon nutrition source.

When creating the invention, the task was to obtain a strain having higher efficiency with high adaptive capabilities.

The problem is solved by a new strain of nodule bacteria Bradyrhizobium japonicum BM-85k, which was isolated in 2003 from nodules of wild soybean that grows on floodplain soils of the Blagoveshchensk district of the Amur Region.

The resulting strain is deposited and stored in the All-Russian collection of microorganisms of the Institute of Biochemistry and Physiology of Microorganisms (IBPM) named after G.K.Skryabin of the Russian Academy of Sciences under registration number VKM B-2456D and the collection of the State Scientific-Research Institute of the All-Russian Research Institute of Soybean under the number BM-85k.

The strain Bradyrhizobium japonicum VKM B-2456D is characterized by the following cultural-morphological and physiological-biochemical characteristics.

Cultural and morphological features

At a young age, the bacteria of the strain Bradyrhizobium japonicum VKM B-2456D are motile, they are elongated rods of size (2.3 ... 2.5) × 0.4 microns. Gram-negative. Fast growing culture. In Petri dishes on mannitol-soy solid nutrient medium, these bacteria grow 20 days after sowing, forming convex, milky-white, mucous colonies with a diameter of 1.0 ... 3.1 mm. The bacteria Bradyrhizobium japonicum VKM B-2456D grow well on other nutrient media:

Norris's medium, g / l: K ₂ HPO ₄ - 0.5; MgSO ₄ - 0.8; NaCl - 0.2; FeCl ₃ - 0.01; yeast extract - 2.0; mannitol - 10.0.

Soybean medium with mannitol, g / l: K ₂ HPO ₄ - 0.5; KH ₂ PO ₄ - 0.5; NaCl - 0.1; MgSO ₄ - 0.1; CaSO ₄ - 0.1; molybdenum salt - traces; mannitol - 20; soy flour - 10.0; agar-agar - 15.0.

Soya medium with glucose, g / l: K ₂ HPO ₄ - 0.5; KH ₂ PO ₄ - 0.5; NaCl - 0.1; MgSO ₄ - 0.1; CaSO ₄ - 0.1; molybdenum salt - traces; glucose - 20.0; soy flour - 10.0; agar-agar - 15.0.

Soybean medium with sucrose, g / l: K ₂ HPO ₄ - 0.5; KH ₂ PO ₄ - 0.5; NaCl - 0.1; MgSO ₄ - 0.1; CaSO ₄ - 0.1; molybdenum salt - traces; sucrose - 20.0; soy flour - 10.0; agar-agar - 15.0.

Soybean medium with maltose, g / l: K ₂ HPO ₄ - 0.5; KH ₂ PO ₄ - 0.5; NaCl - 0.1; MgSO ₄ - 0.1; CaSO ₄ - 0.1; molybdenum salt - traces; maltose - 20.0; soy flour - 10.0; agar-agar - 15.0.

Soybean medium with lactose, g / l: K ₂ HPO ₄ - 0.5; KH ₂ PO ₄ - 0.5; NaCl - 0.1; MgSO ₄ - 0.1; CaSO ₄ - 0.1; molybdenum salt - traces; lactose - 20.0; soy flour - 10.0; agar-agar - 15.0.

A portion of soy flour is obtained by grinding germinated soy seeds, pour 1 liter of water and boil for 30 minutes. Salts are added to the filtrate, adjusted to 1 liter.

When growing VKM strain B-2456D in test tubes on all of the above media (at a temperature of 28 ° C for 480 hours), a marked increase in the stroke of a gel-like milky-white consistency was noted. During long-term storage of pure culture in a refrigerator at a temperature of 4 ° C, the consistency of the stroke of the strain VKM B-2456D does not change.

Physiological and biochemical characteristics

Aerobe.

The temperature range of growth is 27-30 ° C. The optimum growth temperature is 27-28 ° C.

The optimum pH is 7.0.

Relation to carbon sources. It is well absorbed and grows on sucrose, glucose, maltose, lactose, mannitol with the formation of acidic products.

The signs of the strain are resistant. The strain is not pathogenic. Replanted once every 6 months. It is stored in test tubes on mannitol-soy agar at a temperature of 4-5 ° C.

Strain VKM B-2456D was identified by Bergie's “Qualifier” (1980) as a strain of the species Bradyrhizobium japonicum.

The strain Bradyrhizobium japonicum VKM B-2456D differs from the previously known strains of soybean nodule bacteria by its ability to use various sources of carbon nutrition.

The invention is illustrated by the following specific example of the use of a strain of Bradyrhizobium japonicum VKM B-2456D.

Example

Inoculum is obtained by growing the strain in test tubes with agarized nutrient soybean medium with mannitol, g / l: K ₂ HPO ₄ - 0.5; KH ₂ PO ₄ - 0.5; NaCl - 0.1; MgSO ₄ - 0.1; CaSO ₄ - 0.1; molybdenum salt - traces; mannitol - 20.0; soy flour - 10.0; agar-agar - 15.0.

After seeding, tubes with bacterial culture are placed in thermostats with a temperature of 27-28 ° C for 20 days. During this time, in tubes on a jamb of a dense nutrient medium, an abundant increase in the bacterial mass of the test strain is noted.

To inoculate seeds on the day of soybean sowing, a bacterial-molybdenum mixture is prepared. To do this, 2.5 ml of a pre-prepared 1.25% solution of molybdenum (25 g / l ammonium molybdenum) are added to the tubes with a stroke of the test strain of soybean nodule bacteria. The bar from the jamb of the nutrient medium is washed off with a molybdenum solution. By shaking the tubes, a homogeneous bacterial-molybdenum mixture is obtained, which is immediately treated with soybean seeds. The resulting suspension volume (2.5 ml) is used for presowing treatment of 250 g of soybean seeds. After applying the suspension, the soybean seeds are thoroughly mixed, dried in the shade and sown in the prepared soil.

The table provides data on the effectiveness of the strain VKM B-2456D, depending on the conditions of the year, obtained as a result of field experiments. The experiments were carried out on meadow chernozem-like soils with the soybean variety Sonata. Bacterialized seeds are sown on the day of treatment. The strain Bradyrhizobium japonicum VKM B-2456D compared with the control (B. japonicum 648A strain) stably increased the yield of soybean seeds during the three years of testing by an average of 1.8 c / ha.

The effectiveness of the strain Bradyrhizobium japonicum VKM B-2456D Strain Soybean seed yield by years, kg / ha Average productivity, kg / ha 2004 year 2006 year 2007 year Control (strain 648A) 17.1 24.8 16.6 19.5 BM-85k (VKM B-2456D) 18.5 27.4 18.0 21.3

Claims (1)

A strain of soybean nodule bacteria of the species Bradyrhizobium japonicum BKM B-2456D for the manufacture of bacterial fertilizer for soybeans.