RU2415924C1 - Sinorhizobium fredii RNCM NO B-2458D NODULE BACTERIAL STRAIN FOR MANUFACTURING OF BACTERIAL SOYA FERTILISER - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: Sinorhizobium fredii nodule bacterial strain is recovered from a natural population of nodule bacteria common in soils of a southern zone of the Amur Region and deposited in the Russian National Collection of Microorganisms, No. B-2458D, and also stored in the collection of the State Scientific Institution Russian Soya Research Institution, No. BB-49k.

EFFECT: stable improvement of soya seed crop.

1 ex, 1 tbl

Description

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

Known strains of nodule bacteria of soybean 639A, 648A, used for the preparation of soybean nitragin related to the slow-growing species Bradyrhizobium japonicum.

The disadvantages of these strains is that they grow only at pH 7 and a temperature of 27-28 ° C (optimal conditions for growing soybean nodule bacteria), only mannitol is used as a source of carbon nutrition.

The objective of the invention is to obtain a more technologically advanced strain with high adaptive capabilities, the rapid growth of biomass in conditions other than optimal.

The problem is solved by a new strain of nodule bacteria Sinorhizobium fredii BKM B-2458D, which was isolated in 1992 from nodules of wild soybean growing on brown forest 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 BKM B-2458D and in the collection of the GNU of the All-Russian Scientific Research Institute of Soybeans under the number BB-49k.

The strain Sinorhizobium fredii BKM B-2458D is characterized by the following cultural-morphological and physiological-biochemical characteristics. Cultural and morphological characters.

At a young age, the bacteria of the strain Sinorhizobium fredii BKM B-2458D are motile, 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 3-4 days after sowing, forming convex, transparent, mucous colonies with a diameter of 1.0 ... 7.0 mm. The bacteria Sinorhizobium fredii BKM B-2458D 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, obtained by grinding germinated soy seeds, is poured with 1 liter of water and boiled for 30 minutes. Salts are added to the filtrate, adjusted to 1 liter.

When growing the strain BKM B-2458D in test tubes on all of the above media (at a temperature of 28 ° C for 72 hours), an abundant increase in the stroke of a gel-like transparent consistency was noted. During long-term storage of pure culture in the refrigerator at a temperature of 4 ° C, the consistency of the stroke of the strain VKM B-2458D does not change.

Physiological and biochemical characteristics.

Aerobe.

The temperature range of growth is 27-32 ° C. The optimum growth temperature is 27-28 ° C.

Optimum pH 7.0; pH range 6.0-8.0.

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

Osmosis resistant, gives abundant growth on media with a concentration of NaCl of 0.4 microns.

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-2458D was identified by Bergie's Identifier (1980) as a strain of the species Sinorhizobium fredii.

The strain Sinorhizobium fredii VKM B-2458D differs from the previously known strains of soybean nodule bacteria in its ability to use various sources of carbon nutrition and grow well in a wide temperature and pH range.

The invention is illustrated by the following specific example of the use of a strain of Sinorhizobium fredii VKM B-2458D.

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 a bacterial culture are placed in thermostats with a temperature of 27-28 ° C for 5 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-2458D, depending on the conditions of the year, obtained as a result of field experiments.

The effectiveness of the strain Sinorhizobium fredii VKM B-2458D

Strain Soybean seed yield by years, kg / ha Average productivity, kg / ha 2001 year 2002 year 2004 year 2005 year 2006 year Control (no processing) 32,0 26.7 11.9 26.0 18.8 23.1 BB-49k (VKM B-2458D) 32,7 28,2 13.8 27.5 23.5 25.14

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 Sinorhizobium fredii VKM B-2458D compared with the control (background - natural inoculation) stably increased the yield of soybean seeds during 5 years of testing, an average of 2.0 c / ha.

Claims (1)

A strain of soybean nodule bacteria of the species Sinorhizobium fredii BKM B-2458D for the manufacture of bacterial fertilizers for soybeans.