RU1784618C - Strain of bacteria rhizobium sp (ohobrychis) for making fertilizers for sainfoin - Google Patents

Abstract

Usage: microbiological industry, production of means for increasing the yield of leguminous plants, in particular the production of sainfoin fertilizer. SUMMARY OF THE INVENTION strain of bacteria Rhlzobium sp.fOnobrychis) isolated from sainfoin nodules. Deposited in the collection of VNIISCHM under No. 2921. The strain is grown on a medium containing sources of carbon, nitrogen and mineral salts, at aeration and a temperature of 28-30 ° C for 36-48 hours. A culture fluid with a titer of 4-5 billion / ml is obtained, then peat is inoculated, hermetically sealed in plastic bags and cultivate the culture for 3-5 days. at a temperature of 15-20 ° C. Risotorfin is obtained with a bacterium titer of 5-10 billion / g peat. The resulting rhizotorfin increases sainfoin yield by 8.6-26.6%. 2 tab. cl

Description

The invention relates to microbiological means for increasing the yield of leguminous plants,

A known strain of Rhizoblum sp. (Onobrychis) 820, which is used to inoculate sainfoin. However, its nitrogen-fixing activity and manufacturability are not high enough and stable. In addition, crop increases, especially in the southern steppe part of the USSR, are insignificant.

The aim of the invention is to provide a strain of sainfoin nodule bacteria that meets the requirements for the production of rhizotorfin, which has a higher nitrogen fixation activity and improves and improves the quality of the sainfoin crop. As such a strain, strain 2921 is provided.

2921 nodule bacteria isolated from sainfoin nodules in 1985

The strain is determined by the identifier Bergi.

Strain Rhizobium sp (Onobrychis) 2921 was deposited in the collection of the All-Russian Research Institute of Agricultural Microbiology under number 2921.

Strain 2921 grows on nutrient media of the following composition1

1. Pea agar, g / l peas - 100 0 sucrose - 20.0; agar - 20.0, pH 7.0-7.2

2. Liquid medium Norris g / l K2NRO / 1 0.5; MgS04-7H20 - 0.8, FeCl3 - 0 01 mannitol - 10.0, yeast extract - 100 ml

pH 7.1.

Morphological and cultural properties Vegetative cells

00

On pea agar with a pH of 7.1–7.2 at 28–30 ° C in a six-day-old culture, they are in the form of rods 1-2–0.8–0.6–0.7 μm in size, gram-negative, type of towing - perichi. In young cultures, they are motile, homogeneous, and with aging, the hbnB is replaced by granularity with a simultaneous loss of mobility. During aging, the culot $ s form bacteroids, the image of which is $ bp large flask-like: lettuce, .J

Characterization of growth in a dense medium.

Pea agar, T 28-30 ° C, pH 7.0-7.2

Colonies appear on the sixth day, mucous, translucent, convex, round, sometimes irregular in shape, up to 2 mm in diameter.

Growth on carbohydrate media is usually accompanied by the formation of abundant extracellular polysaccharide mucus.

Physiological and biochemical properties.

Symbiont.

Macrosymbiont is a sainfoin bean plant.

Kg ratio

a) oxygen - aerob

b) T ° maximum 40 ° C

minimum 10 ° C

optimal - 27-29 ° C

c) pH maximum 7.5

minimum 6.0 is optimal. 7.0-7.2

9th

GlucoseAsch (-)

LactoseAsch (-)

MaltoseAsch (-)

XyloseAsch (-)

SucroseAsch (-)

RamnoseAsch (-)

MannitAsh (-)

SorbitolAsc (-)

Cellulose-

Starch

Liquid medium Norris, T 26-28 ° С Sources of nitrogen

W NaNOa

(NH4) 2 $ 04

tyrosine

MPA

Nitrogen is fixed in symbiosis with esp arcem.

Other characteristic physiological features.

It does not dilute gelatin.

+++

Milk with litmus does not peptonize, alkalizes, forming the mucous zone on the surface.

The content of G + C in DNA is 62.3%.

The strain is not pathogenic. Strain 2921 was stored on pea agar of the above composition at 2-4 ° C for 6 months, then reseeded.

PRI me R 1. Cooking techniques

drug based on strain 2921.

The seed culture is grown on a rocking chair in flasks or five-liter bottles for 4 days on a medium of the following composition, wt.%: Pea broth - 10.0; sugar (food) - 1.0: (MNfZSm - 0.05; KN2P04 - 0.05; K2NRSM-ZN20 - 0.05; MgSCM 7H20-0.02; CaCO 3-0.1; tap water - up to 100, 0. pH after sterilization (1.0 these, 30 min.) - 6.8-7.0.

cells 3-4 billion / ml. A seed crop is used to prepare a liquid work crop. It is ready for T1 in five-liter bottles or fermenters in an environment of the following composition, wt.%: Corn

extract - 1.0; molasses - 2.0; (MH4) 25Sm - 0.1: KH2P04 - 0.025; CaNRSm-ZNaO - 0.025; Md5Sm-7N20-0.02; CaCO3 - 0.1; tap water up to 100.0. pH after sterilization of 6.8-7.0.

Bottles and fermenters are seeded with a seed culture of nodule bacteria in an amount of 5%. The bottles are shaken on a rocking chair for 80-96 hours. In farms, bacteria grow 36-48 hours at

aeration of 0.7-1.5 r / g.in under an excess pressure of 0.3-0.5 these at a temperature of 28-30 ° C, a titer of 4-5 billion / md. After microbiological control, the culture fluid from inoculum bottles or fermenters is used for mixing with peat. Peat preparation.

Ground lowland peat neutralized with chalk to pY 6.8-7.0 with a moisture content of 30-40% in plastic bags of 200400 g is sterilized with gamma irradiation at a dose of 2.5 Mrad. Sterile peat in bags is inoculated with an injection of a mixture of culture fluid with molasses (6%) based on the initial titer of bacteria in peat

0.3-0.5 billion / g of peat and humidity of 45-60%. The bags are then loaded into special mixers, the rotation of which mixes their contents. Culture c. peat is grown by aging packages

3-5 days at a temperature of 15-20 ° C. Risotorfin is stored at a temperature of 3-8 ° C in a dark, dry room, separate from the pre-chemicals. Characteristics of the finished product.

Rizotorfin on the proposed strain is in appearance a moistened loose mass of dark color, odorless, insoluble in water. The humidity of the finished product is 50-55%. Rizotorfin contains 5-10 billion nodule bacteria in 1 g of peat. The shelf life of the drug is 6 months from the date of manufacture. The consumption of the drug is 200 g per hectare norm of seeds.

EXAMPLE 2 A comparative study of strains of Rhlzobtum sp. (0nobrychls) 820 and 2921 was carried out in vegetative and daily experiments with sainfoin of the South Ukrainian variety. In the growing experiments, dark chestnut soil was used containing 2.7 mg hydrolyzable nitrogen per 100 g of soil. Weighed soil in vessels of 6 kg. Repeatability is ten times. Business experiments were carried out on meadow chernozem-like soil with a content of easily hydrolyzable nitrogen 5.1 mg per 100 g of soil. The size of the accounting records is 2 m2, the repetition is four times.

The results of the experiments are given in table 1.

As can be seen from Table 1, the increase in green mass yield from inoculation of sainfoin seeds with strain 2921 was 60.0 c / ha (15.4%) compared with the uninoculated variant. Strain 820 was less effective.

Example 3. The same strains were tested under production conditions on sainfoin of the Yuzhnoukrainsky variety on southern loamy chernozem with easily hydrolyzable nitrogen soderonium 6.1 / 100 g of soil. The size of the accounting records is 50 m2, the repetition is fourfold.

The results are shown in table.2.

As can be seen from Table 2, strain 2921 is more effective compared to the prototype strain 820 and in the field.

The proposed strain, in addition, increases the protein content in the green mass of sainfoin by 1.8%, which increases its feed value, as well as high-tech - gaining a titer of up to 10 billion cells in 1 g of rhizotorfin.

FORMULA AND SECTION

The bacterial strain Rhizobium sp. (Onobrychis) VNIISCHM Mg 2921 for the production of sainfoin fertilizer.

Table 1

The effectiveness of inoculation of seeds of sainfoin with risotorfin, made on the basis of

strains 820 and 2921

1784618

8 table 2