SU1446132A1 - Strain of rhilobium leguminosarum tuber bacteria for producing bacterial fertilizer for lens - Google Patents

Abstract

This invention relates to microbiological means of increasing leguminous crops and can be used to produce nitragin fertilizer for lentils. The aim of the invention is to obtain a new, more efficient strain of nodule bihisteries, which produce grain yield and protein content. The strain of nodule bacteria Rhizobium legu-rainosarum 724 was isolated from nodules of lentil varieties Petrovsk 4/105. The strain of nodule bacteria of the lentil Rhizobium leguminosarxim (Lens) 724 was deposited and stored in this collection in the ARRIEM collection under No. 724. When the lentils were inoculated with the strain Rhizobium legumino-sarum (Lens) 724, the yield of green mass increased in comparison with the variant inoculated with the R. rum strain of the strain Rhizobium legumino-sarum (Lens) 724. a, at 0.4 g / vessel (30.8%) and the total nitrogen content in plants by 0.31% (vegetation experience), compared to uninoculated pzb-bi many lentils, the grain yield in field experiments is average 9.5 centners per hectare (79.8%) and protein content in grain by 0.9%. 2 tab.

Description

The invention relates to microbiological means of increasing the yield of leguminous crops and can be used for the production of bacterial fertilizer nitragin for lentils,

The aim of the invention is to obtain a new, more efficient strain of nodule bacteria, which increases the yield of grain of lentils and its protein content.

Strain nodule bacteria Rh. leguminosarum (Lens) 724 is grown from nodules of Lentil A / Yu5 Petrovsk variety grown on ordinary black soil.

 Identified by Berg.

Strain nodule bacteria Rh. legiBninos arum (Lens) 724 is deposited and stored in the VNIISM collection under 724.

Cultural and morphological properties of the strain Rh.leg. (Lens) ARRIAM 8 724.

Calculation on the trace of nutrient media.

Pea agar, g / l: pea 100.0; sucrose 20.0; agar-agar 20.0; pH of 6.8-7.0. On the third day, colonies on pea agar appeared of the same type, round, whitish, strongly convex, 2-2.5 mm in diameter.

Bacteria have chopsticks that are motile at a young age. The size of the cells of a 2-day culture on pea agar 1.2-2.5 0.6-0.9 MCO

Peritrichs Bactero; ides are club-shaped. Stroke on pea agar - plentiful, whitish, slightly weak, mucous

Lupine agar: g / l: K NRO 0.5; KHjPO 0.5; MgSO -THiO 0.2; NaCl 0.2; CaS04 0.1; (NH) iMo04 traces; mannitol 20.0; lupine flour 10.0; agar-agar 20.0, pH. 6.8-7.0.

The stroke is plentiful, whitish, slightly convex, slimy, and stacked. Colonies appear on the third day, of the same type, round, whitish, convex 2.5-3.0 mm in diameter.

Mannitol-yeast agar, g / l: mannitol 10.0; 0.4; agar-agar 15.0 pH of 6.8-7.0.

The stroke is plentiful, colorless, flat, slimy, strongly flowing. Colonies occur on the third day, same round, whitish, convex

0

five

0

five

0

five

spreading, 3-3.5 mm in diameter.

Physiological and biochemical characteristics of the strain Rh.leguminosarum (Lens) 724.

Milk with a litmus does not peptonize, alkalinize with the formation of a mucous ring on the surface. Gelatin does not dilute. Aerobe, Optimum temperature 26-28 C.

Attitude to carbohydrates “Grows on media with carbohydrates, acidic medium. It develops well on media with glucose, galactose, rhamnose, xylose, less actively on media with maltose, sucrose, lactose. Fiber and starch do not. Digest.

Relation to nitrogen sources. It grows well on media with sodium nitrate, ammonium salts, tyrosine, MPA does not grow. Signs of the strain are resistant Strain is not pathogenic. It is subcultured once every 6 months and stored on pea agar at 4-5 ° C.

The method of preparation for the inoculation of lentils in field and vegetative experiments. Preparation of liquid seed culture. Liquid culture is grown in rocker flasks and in 5 liter bottles for 2 days on the medium of the following composition,%:

Pea decoction 10.0 Sugar (food grade) 1.0 (Ш4) г5040.05

КН2.Р040,05

Kj HP04-3H2.00,05

MgS04, 02

CaCOj0,1

Water supply one

waterUn.100

pH (after sterilization) 6.8-7.0 Sterilization at 1.0 atom and 30 min. Title 5-8 billion / ml. The seed culture is used to prepare a liquid working culture in 5 liter bottles or fermenters on a medium of the following composition:%:

0

five

Corn

extract

Molasses

(NH4) tS04

KHjP04

Kz.HP04 3HiO

MgS04-7HiO

CaCOj

Water pipe

water

1.0

2.0

0.1

0.025

0.025

0.02

0.1

Up to 00.0

one

pH (after sterilization) 6.8-7.0

The bottles and fermenters are seeded with 5% seed culture of nodule bacteria. The bottles are shaken on a rocking chair for 40-48 hours. In the fermenters, bacteria are grown for 18-36 hours with aeration of 0.7-1.5 rpm under an overpressure of 0.3-0.5 to 28-30 C. Titer not less than 5.0 billion / ml. After microbiological control, culture liquid from bottles or fermenters (inoculum) is used for mixing with peat.

Peat preparation. Ground ground peat neutralized with chalk to a pH of 6.8-7.2 with a moisture content of 30-40% in plastic bags up to 200-400 g is sterilized with gamma irradiation at a dose of 2.5 Mrad. Sterile in packets is inoculated by injecting the mixture: cultural liquid + molasses (6%) based on the initial titer of bacteria in peat 0.3-0.5 billion / g peat and humidity 45-60%. The bags are then loaded into special mixers, the rotation of which mixes their contents.

The culture in peat is grown by keeping the bags for 3-5 days at 15 - 20 ° C, then the bags are stored at 3 - 8 ° C in a dark, dry room, separate from the chemicals.

Characteristics of the finished product.

Rhisotorphin for lentils on the proposed strain is in appearance a moist, dark, odorless bulk, insoluble in water. The moisture content of the finished product 50-55% o Risotorphin contains 4-6 billion, nodule bacteria per 1 g of peat. The shelf life of the drug is 6 months from the date of manufacture. Consumption of the drug 200 g per hectare rate of seeds.

In vegetation experiments, inoculation with lentils was inoculated with a suspension of a pure bacterial culture, washed off agar, in field trials with a peat preparation, rhisotorphin.

The preparation for mastering the strain Rhizobium leguminosarum (Lens) 724 for inoculation of lentils is used in the following way.

Example 1. Vegetation experiments were carried out in vessels with 6 kg of sod-podzolic soil with the addition of salts.

32

According to the recipe Pr Nishnikova, without nitrogen. 4-fold repetition of experiments “Inoculate seeds with Penzenska 14 varieties of lentil strains,

The results of vegetation experiments are shown in table 1.

As the data of Table 1 show, bacteria of strain 724 actively fix atmospheric nitrogen in symbiosis with lentils, as a result of which the yield of green mass of plants bacterized with this strain is 53-107% higher than that of plants without inoculation, while increasing total nitrogen by 0.34-0.70%. As a result, the total accumulation of nitrogen in inoculated plants is 2.3–2.6 times higher than non-inoculated ones.

In comparison with the known strainer rhizobia 250a, strain 724 increases the yield of green mass of plants by 0, -4 g / vessel (30.8%) and the total nitrogen content in lentils by 0.31%.

PRI mme R 2. Field experiments carried out on ordinary chernozem. The area of the delta is 25 m. The repetition of the experiments is 4 times. Preparation for inoculation of lentils of two varieties of Penzensk 14 and Petrovsk 4/105.

The results of field experiments are given in table 2.

As can be seen from Table 2, the R.legu minosarum (Lens) 724 strain produced an effective symbiotic nitrogen fixer under field conditions. The yield of lentil grain when inoculated with this strain increases on average by 9.5 q / ha or by 79.8%, the protein content in the grain by 0.9%.

Thus, when lentils were inoculated with R. legximinosarum (Lens) 724 strain, the yield of green mass increased compared with that of strain 250a by 0.4 g / container (30.8%) and the total nitrogen content in the plant was 0.31%. In comparison with non-inoculated lentils, the grain yield increased by an average of 9.5 centners / ha (79.8%) and the protein content in the grain increased by 0.9%.

Claims (1)

Invention Formula Strain of nodule bacteria Rhizo bium leguminosarum (Lens) ARRIAM. No. 724 for the production of bacterial fertilizer for lentils. Table I  Wore inoculum I Without inoculum Table 2