Spectrophotometric indicators of “Zamin-M” biopreparation immobilized with gypan flocculinate

. From the turn of the 20th century, biopreparations that quicken plant growth and development have been frequently employed. The accumulated knowledge of the mechanisms governing interactions between microorganisms and plants necessitates the use of minimal resources and energy in the targeted design of phytomicrobial systems and the optimization of their adaptability in order to increase soil fertility and plant productivity, increase plant yield, and increase plant resistance to diseases and unfavorable environmental conditions and stress factors. In the article today, enough experience has been collected on the scientific management of soil microorganism processes in the optimization of agricultural production and maintaining soil fertility, and it envisages the creation and use of microbiological preparations as the main link of intensive technologies in plant science. It is known that concentration of biomass of microorganisms using flocculants in the technological processes of production of bacterial preparations, after the stage of growth in enzymes, the stage of concentration of biomass of microorganisms from culture liquid is carried out.


Introduction
The use of microorganisms that accelerate the growth and development of plants as biopreparations has been widely used since the last century [1]. The accumulated knowledge about the mechanisms of interactions between microorganisms and plants requires the use of minimal resource and energy costs in the targeted design of phytomicrobial systems and the optimization of their adaptability in order to increase soil fertility and plant productivity, and increase plant yield, resistance to diseases and adverse conditions, and stress factors [2].
In general, rhizobacteria are one of the potential and brilliant tools in creating sustainable agricultural practices and maintaining existing traditions. Therefore, it is necessary to find out whether it is possible to use the beneficial properties of any bacteria, taking into account the various conditions of the environment and the characteristics of plants. Development of dry powder forms of microbiological preparations [3]. According to the type of product, biological drugs can be divided into two in the biotechnological diagram: 1. Drugs based on living cells; 2. Preparations based on secondary metabolites of microorganisms [42].
The vacuum-sublimation dehydration method is used to obtain the dry form of biopreparations used in various branches of the microbiological industry. This method makes it possible to obtain a high-quality product [5].
"Organika-S" preparation Bacillus amylolique faciens VKPM V-12464 consisting of a strain of It was grown in a nutrient medium containing bacteria of the genus Bacillus . The biomass of rhizobacteria was obtained by cultivation in meat peptone broth [6][7][8][9].
Antigonistic activity of the drug against the main phytopathogens of cotton in the following mycelial microscopic fungal strains: Fusarium oxysporum, Alternaria alternata, Rhizoctonia solani JGKuhn. was seen. All strains of micromycetes are stored in ToshDAU collection of microorganisms [10].
Within this study, the method for producing "Zamin-M" biopreparation in dry form that is immobilized with flocculant has been devised and put into practice. The capacity of soil rhizobacteria to speed up plant growth and development under salt stress has proven extremely beneficial for the long-term sustainability of agriculture around the world. The dry form of the biopreparation was found to belong to the genus and was chosen as the basis for the creation of stimulant drugs for further research. Improving the production technology of the dry powder form of the biopreparation "Zamin-M". Extraction of concentrated microbial biomass from the culture liquid. Development of the optimized technology for obtaining the immobilized form of the biopreparation.
Gypan is a partially hydrolyzable polyacrylonitrile that is highly soluble in water. A solution of 9-12% in water is usually used. The color varies from light yellow to dark brown. The density at -20 0 C was 1.06-1.07 g/cm.. It does not freeze until 5-10 in the cold.

Results and discussion
Our studies on the concentration of the biomass of microorganisms were carried out by applying Gypan polyacrylonitrile in the proportions of 0.06%, 0.13%, 0.2%, 0.27% to the culture fluids of the rhizobacterial strains that make up the biopreparation "Zamin-M" .
The advantage of this polymer is that it takes place when the conditions of polymerization are рN-7-8, and it is explained by the fact that it corresponds to the conditions of development of rhizobacteria strains that are part of the biopreparation "Zamin-M" adapted to the conditions of salinity stress in an alkaline environment. Another advantage of the polymer is that it does not freeze at a temperature of 5-10 0 C, which allows to keep living cells viable for a long time. The values of the optical densities of bacterial cells concentrated with Gypan below are shown in Figure 1.

Fig. 1. Optical density of bacterial cells concentrated with Gypan
The density of the supernatant of cells precipitated with a concentration of Gypan of 0.1% was 0.070968 mg/l, 0.092487 at 0.2% Gypan, 0.97640 at 03%, 0.81941 at 04%, It was noted that it reached 1.06351 mg/l at 0.5%, that 0.1% Gypan was the reason for the clarification of the settling liquid, that it was difficult for biomass to settle in other parameters, that increasing the concentration of Gypan causes the polymer to thicken with biomass ( Figure 2). After that, it was decided that the concentration of Gypan should be reduced. Precipitation of the biomass of microorganisms included in the "Zamin-M" biopreparation from the culture liquid under the influence of coagulant was used. As can be seen from Figure 3, the relative density of biomass in the sedimentation liquid in the experiment with a concentration of gypan of 0.01% is 0.53283 mg/l, and in the experiment with 0.06% gypan, this indicator is 0.09687, compared to the first experiment . almost 5.5

Gypan relative concentration
Relative density times, 9.9 times less compared to 3 experimental options (gypan 0.1%), 5.53 times less than 4 experimental options (gypan -0.4%), less than 5 experimental variants (gypan 0 .5%) was 5.61 times lower, which indicated that it is superior in its ability to clarify the sediment surface liquid and adsorb more. Comparing the initial experiment and the subsequent experiment presented in Figure 3, the supernatant density of cells precipitated with a concentration of Gypan of 0.1% was 0.070968 mg/l, 0.06% Gypan was used. in the experimental version, this indicator was 0.09687 mg/l and was higher by 0.026 mg/l, so the 0.1% concentration of Gypan was found to be the optimal option. When the number of cells precipitated with Gypan polymer was studied, it was noted that the cell viability was the highest at a concentration of 0.1% [9,10] ( Table 1). The flocculant was diluted with 2-7 volumes of water to introduce it into the KS. Then the mixture of KS and flocculant was stirred for 2 minutes for complete mixing of the components of the solution and coalescence of the fragments. Precipitation was carried out at room temperature, pH of the culture fluid was 9.0. The optical density of the culture liquid was determined relative to the control centrifuged supernatant. The results of the experiment show that biomass sedimentation changes significantly at different values of the concentration of the Gypan reagent. This may be due to an increase in the viscosity of the culture liquid, which did not lead to the sedimentation of the formed floccules, or to the charges or morphological characteristics of the microorganism cells, or to the composition, physico-chemical properties of the culture liquid.
The optimum concentration of flocculant was found to be 0.06% flocculant applied to culture fluid for 90% sedimentation of KS. Biomass sedimentation period was 90 minutes. When examining the morphological signs of the association of microorganisms, it was noted that deposition did not affect the changes in the cultural-morphological signs of microorganisms in the biopreparation, and the ratio of microorganism strains Ps stutzeri SKB 308, B. subtilis SKB -309, B. megaterium SKB 310 was preserved in a ratio of 1:1:1 [10].
Thus, when the flocculation process was used for the "Zamin-M" association of microorganisms, the concentration of the flocculant in relation to the culture liquid was 0.1% by volume and the length of the sedimentation period was 90 minutes, resulting in 90% biomass sedimentation. It was achieved to preserve the proportion, stability and viability of microorganisms in the sedimented biomass .
"Zamin-M" biopreparation in the form of dry powder in accordance with the relevant technical conditions depends on the plant variety and indigenous microflora grown in specific soil and climate conditions.
The working solution is applied to the warmed seeds by drop spraying at 10 ml/kg of seed. It is carried out in a cool place with an air temperature of 18 o C to 30 o C (without direct sunlight). The shelf life of the drug is 2 years from the time of preparation at a temperature of 5 o C +25 o C.
The stage of preparation of the dry form of the biopreparation "Zamin-M" consisted of the following (Figure 4). Preparation of nutrient medium: the nutrient medium is sterilized in an autoclave at 120 o C at 0.1 MPa atmospheric pressure for 40-60 minutes. Planting of inoculum: the inoculum consisting of an association of microorganisms was planted in 20 l bottles, grown at 28 o C for 3 days under conditions of aeration consumption of 3.5 l/min. Sterilization of the technological system is carried out with steam at 120 o C and 0.3 MPa pressure for 40-60 minutes. It was carried out in 2 industrial fermenters with a volume of 3.2-6.3 m 3 for 72 hours at 28 o C. Biomass concentration of microorganisms was carried out in a 4 m 3 reactor using Gypan flocculant. The concentration of the reagent is 0.1% by volume relative to the culture liquid. The mixing period of biomass with flocculant is 1 hour, and the sedimentation period is 12 hours. Extraction of biomass from culture fluid should be observed. Drying the biomass and mixing it with kaolin in a ratio of 100:5. Biomass was dried in a vacuum dryer with a pressure of 0.7 MPa and a loading volume of -0.4 m 3 at 30 o C. The duration of drying was 24 hours [3,9]. According to the produced technical conditions, an experimental sample was developed and tested in a vegetative and microvegetative experiment.

Conclusions
Globally, salinity is one of the major threats to agricultural production. Nevertheless, the issue of using saline lands to meet the food needs of the growing population is urgent , and the viability and functional properties of immobilized cells have been determined.
The technology of obtaining the dry preparation form of "Zamin-M" biopreparation immobilized with flocculant has been developed and implemented. The ability of soil rhizobacteria to accelerate the growth and development of plants in conditions of salinity stress has been providing great benefits in the field of sustainable development of agriculture in the whole world. Extraction of concentrated microbial biomass from the culture liquid, development of the optimized technology for obtaining the immobilized form of the biopreparation, and the dry form of the biopreparation was determined to belong to the genus and was selected as the basis for the creation of stimulant drugs for further research , Improving the production technology of the dry powder form of the biopreparation "Zamin-M" immobilized with Gypan flocculant will lead to great results.