RU2126050C1 - Method of submerged culturing of microbial association on bases of earthworm - producer of phytohormones - Google Patents

Abstract

FIELD: biotechnology. SUBSTANCE: invention relates to preparing phytohormone complex of broad spectrum by using a product of worms Eizemia foetida product vital activity as a substrate. Cultivation is carried out by mixing 50 vol.% of earthworm product vital activity with distilled water at 26-28 C under aeration at intensity 0.15-0.5 vol. rev./min. Process is carried out at medium pH = 6.0-7.8 for 116-124 h. Ready product contains indolyl-3-acetic acid, isopentenyladenosine, gelatin, humic substances and trace elements. Preparation is used for seed treatment before sowing and vegetating plants. EFFECT: enhanced effectiveness of plant growing. 2 cl

Description

 The invention relates to microbiology, in particular to the use of microorganisms to obtain enzymes and phytohormones.

 There are various methods of cultivating microorganisms, in particular fungi and bacteria, used to extract enzymes, vitamins and phytohormones from them. For example, the isolation and cultivation of a culture of microorganisms, the preparation of a nutrient medium, plating the culture on a nutrient medium, fermentation, separation of metabolic products.

 The physiological characteristics of the life of microorganisms and the ability of some of them to super-synthesize metabolic products led to their use for the microbiological synthesis of enzymes and phytohormones. For example, it is known to use Fusarium moniliforme fungi for the biosynthesis of gibberellins (ed. St. SU 1768633, class C 12 N 1/14), Aspergillus niger fungi for the production of hydroxy derivatives of indolyl-3-acetic acid (ed. St. SU 994558, class. C 12 P 1/02). The bacterial product is L-tryptophan (ed. St. SU 990814, class C 12 P 13/22).

 The nutrient medium for microbiological synthesis in a large number of cases is an aqueous mixture or suspension of organic compounds or substances and mineral salts. Various organic substances and compounds can be used to prepare nutrient media: from sugar and starch (in laboratory conditions) to wood hydrolysates, petrochemical wastes and food industry (in production conditions). To these sources of organic compounds are added mineral salts of potassium, magnesium, iron, sodium, calcium, etc. in a calculated amount that meets the needs of microorganisms.

 However, the disadvantages of these methods are the narrow focus of their productivity, the result of which is the production of substances of one or two names, and the complexity of preparatory operations, such as the isolation and cultivation of a culture of microorganisms and the preparation of a nutrient medium.

 There is also known a method for producing biological complexes using liquid from the digestive tract of lower invertebrates as a source of microorganisms, where the latter are subsequently grown on nutrient media (FR patent 2174659, CL 12 P 1/04, 11/23/73).

 However, this method has disadvantages associated with the isolation and cultivation of a culture of microorganisms and the preparation of a nutrient medium. In addition, the disadvantage of this method is the presence of a procedure for selecting fluid from the intestines of larvae or worms.

 The purpose of the invention is the elimination of these disadvantages by obtaining a complex of phytohormones of a wide spectrum by using Eizenia foetida as a source of vital activity of microorganisms and a source of nutrients.

 The proposed method of deep cultivation of the source of the association of microorganisms is as follows.

The product of the vital activity of earthworms Eizenia foetida in an amount of 640 l is mixed with 1280 l of distilled water and fermented for 120 hours at a temperature of 28 ^o C. The mixture is continuously aerated with air, and the pH is maintained at 7.8. The intensity of aeration is 0.15-0.5 rpm / min.

 The vital product of earthworms Eizenia foetida contains a wide range of microorganisms - producers of phytohormones, nitrogen-containing nutrients and a wide range of trace elements.

 Fusarium moniliforme fungi are synthesizers of gibberellin.

 Aspergillus niger fungi produce hydroxy derivatives of indolyl-3-acetic acid and humic substances.

 The bacteria Corynebacterium glutamicum and Bacillus subtillis produce L-tryptophan, which is subsequently transformed into heteroauxin (β-indolylacetic acid).

 The bacteria Azotobacterium chroococcum are producers of heteroauxin and gibberellin. With further use in the complex microfertilizer (CMU) obtained by the proposed method, for the treatment of crops of crops, Azotobacterium chroococcum forms a nitrogen-fixing association of microorganisms on the plant.

Bacteria of Rhizobium as part of the microbiological community of the indicated CMU, when processing plants, form colonies of nodule bacteria on their roots that provide nitrogen nutrition. The species of cultivated bacteria is determined by the composition of the manure used to feed the worms. So, if the clover red was part of the animal feed, then in the process of deep cultivation, the bacteria Rhizobium trifolii are predominantly developed, further respectively:
alfalfa - Rh. meliloti;
soybeans - Rh. japonicum;
peas, vetch, lentils - Rh. leguminosarum;
Lupine - Rh. lupini;
Beans - Rh. phaseoli;
sainfoin - Rh. onobzychis.

 Bacillus megaterium bacteria, which are part of the microbiological community of CMU, when applied to plants, provide their phosphorus nutrition.

 The livelihood product of the earthworm Eizenia foetida is 48% organic matter. In addition, it includes mineral elements: potassium, calcium, magnesium, sulfur, manganese, zinc, copper. Their quantitative content determines the ratio when mixing the product of the vital activity of worms with water in the proposed method.

The microorganisms listed above are aerobes, therefore, cultivation is carried out with constant saturation of the mixture with oxygen in the air, blown through the mixture with an intensity of 0.15-0.5 rpm. The temperature of the mixture is maintained at 26-28 ^o C and is optimal for the life of bacteria and fungi. The hydrogen index of the mixture is maintained at 6.0-7.8 for the same reasons. The full cycle of microbiological synthesis in the proposed method is carried out in 116-124 hours

 Thus, the use of Eizenia foetida earthworms as a source of nutrient medium and microfloral association for deep cultivation of the vital product of earthworms allows one to obtain CMU containing plant growth regulators, nutrients, a wide range of microelements, and soil complex microorganisms.

 The use of complex micronutrient obtained by the proposed method for processing seeds before sowing and vegetative plants can increase crop yields by 10-25%.

Claims (2)

 1. The method of deep cultivation of a source of microbial association - a producer of phytohormones from earthworms by introducing it into a nutrient medium and aeration, characterized in that the source of microbial activity of earthworms Eizenia foetida is used in an amount of 50% by volume of the medium, cultivation is carried out on distilled water with aeration intensity of 0.15 - 0.5 rpm / min.  2. The method according to claim 1, characterized in that the pH of the medium is maintained at 6.0 - 7.8.