RU2043028C1 - Method for production stimulant of growing plants - Google Patents

Abstract

FIELD: agriculture. SUBSTANCE: before extraction culture liquid which contains mycelium of fungus endophyte is treated with the help of ultrasound, its intensity being 0,3-1,2 V/cm². Said process is carried out within 10-20 min. It is followed by drying, dry precipitate of culture liquid being extracted. EFFECT: improves efficiency of the method. 3 cl, 6 tbl

Description

 The invention relates to agriculture and relates to a method for producing a plant growth stimulator of the Symbiont type, which can be used to increase the yield of various agricultural plants (potatoes, cucumbers, lettuce, etc.).

A known method of obtaining the drug "Symbiont-1", which consists in the following. The endophyte fungus isolated from ginseng roots is cultivated on a nutrient medium containing glucose as a carbon source, asparagine as a nitrogen source, as well as sources of phosphorus, potassium, magnesium, iron, manganese. Culturing is carried out in optimal conditions of growth of the fungus, for example at 28 ^{° C} for 30 days. Sterile activated carbon is introduced into the obtained culture fluid containing fungal mycelium, which adsorbs biologically active substances (active principles) from the culture fluid and mycelium, after three hours the charcoal is filtered off, the filter cake is washed with 70% ethanol. Ethanol extract is used as a preparation "Symbiont-1". The amount of ethanol used for extraction should be such that the volume of the preparation obtained is 1/3 of the volume of the culture fluid. When using the drug "Symbiont-1" the yield of cucumbers, oats, barley increased by 12.3-19.9% and potatoes up to 65%, "Symbiont-1" is used to process both seed and growing plants. The disadvantage of this method is the scarcity of raw materials from which the initial mushroom endophyte is obtained, which prevents the organization of mass production of Symbiont-1.

 There is another way to obtain a plant growth stimulator, called "Symbiont-2", which consists in the fact that sea buckthorn roots are used as raw material for the isolation of the original endophyte fungus. The technology for producing a growth stimulator is identical to that described above. The growth stimulator obtained by this method can be used when diluted 10 thousand times for pre-sowing treatment of plant seeds and, when diluted 50 thousand 100 thousand times, for introduction into the soil with irrigation water.

 The disadvantages of the methods is the relatively low activity of the target product and the associated high consumption of stimulant per unit area.

 The basis of the invention is the task of developing a method for producing a plant growth stimulator of the Symbiont type, which provides high activity of the target product.

The problem is solved as follows. The endophyte fungus isolated from any raw material, in particular from sea buckthorn, half a floor, is cultivated under optimal conditions on a nutrient medium containing sources of carbon, nitrogen, phosphorus, potassium, magnesium, iron, manganese, for 1-6 months. Temperature 20-25 ^{° C,} pH 6.5. The culture fluid containing mycelium is treated with ultrasound with an intensity of 0.3-1.2 W / cm ² for 10-20 minutes. The resulting mixture was dried at a temperature of 20-40 ^C. The dry residue is treated with 40-70% ethanol to extract active principles. The extract is filtered and the filtrate after appropriate dilution is used for pre-sowing treatment of seeds of cucumbers, lettuce, etc. and for introduction into the soil with irrigation water. The specified filtrate is a symbiont type plant growth promoter, which is called “Symbiont K”. Its activity is 5-20 times higher than the activity of Symbiont-2 and Symbiont-2 preparations.

 Ultrasonic treatment of the culture fluid with fungal mycelium causes destruction of the cell walls of the mycelium of the endophyte fungus and promotes the release of intracellular biologically active substances into the culture fluid. The choice of ultrasound parameters and the duration of ultrasonic treatment of the culture fluid is determined by expediency. It was found that when the parameters are lower than the specified lower limits, the completeness of the extraction of biologically active substances from the mycelium is not achieved. At higher values of the parameters than the indicated upper limits, a difficult to filter dispersion is obtained, which complicates and increases the cost of obtaining the target product.

 Growth stimulator activity is measured by the rate of oxygen uptake in the soil with the roots of 7-day-old cucumber seedlings developed in it, the seeds of which were previously treated with a growth stimulator. A significant increase in the rate of oxygen uptake compared to the control indicates an increase in the synthesis processes in the roots and correlates with the future crop.

 The rate of oxygen absorption is measured by the amount of absorbed gas, reduced to normal conditions and referred to 1 kg of soil and 1 h.

PRI me R 1. The culture of the endophyte fungus isolated from the roots of sea buckthorn, cultivated on a nutrient medium composition, g: glucose 8; K ₂ HPO ₄ 0.6; KH ₂ PO ₄ 1.8; MgSO ₄ 0.2; K ₂ SO ₄ 0.1; MnSO ₄ and FeSO ₄ traces; Asparagine 20 mg per 1 liter of tap water at a temperature of 20-25 ^{° C,} pH 6.5, for 2 months.

The culture fluid containing mycelium, endophyte fungus is treated with ultrasound with a wavelength of 59.2 mm and an intensity of 0.3 W / cm ² in an ultrasonic cleaner UM-4 "Winta" of Polish production for 20 minutes.

 Then the resulting dispersion is dried at room temperature, the dry residue is treated with 40-70% ethanol, the mixture is kept for three hours and filtered. The filter residue is washed with 40-70% ethanol. The total amount of ethanol is 1/3 of the volume of the original culture fluid.

 The resulting alcoholic extract is a symbiont-2K plant growth stimulator. Its activity was determined as follows. The drug is diluted 100 thousand or more times. For comparison, use a standard (diluted 10 thousand times) solution of Symbiont-2. The tests were carried out with a hybrid of cucumbers "Regatta" and soils from greenhouses. The seeds were soaked for 30 minutes with the preparations: one portion of Symbiont-2, the second portion with a new preparation. The control portion was soaked in distilled water.

To measure the rate of oxygen absorption, a Warburg apparatus was used. The reliability of the results was checked by pairwise comparing the results of the options with the control. It is expressed by the actual value of the student criterion (t _f ) and compared with the theoretical value of the student criterion (tm) at significance levels of 0.00 and 0.01.

 Comparative data on the activity of the drugs are presented in table 1.

 As from Table 1, the Symbiont-2K preparation obtained according to the invention, being diluted 10-20 times more than Symbiont-2, provides a higher oxygen absorption compared to the control, i.e. the activity of the obtained growth stimulator is 10-20 times higher than the prototype.

 PRI me R 2. The method is carried out analogously to example 1, but using an endophyte fungus isolated from half the floor. The resulting preparation is called "Symbiont-3K." The results were compared with a drug called "Symbiont-3", the manufacturing technology of which does not involve the use of ultrasound. The results are presented in table.2. The significance of the difference (in terms of tf) is given in comparison with the control.

 As can be seen from the table. 2, the activity of the drug "Symbiont-3K" is 5 times higher than the activity of "Symbiont-3" obtained in a known manner.

 In the table. 3-5, information is presented that substantiates the parameters of ultrasonic treatment of the culture fluid.

As follows from table 3, the absorption of oxygen is maximum when the ultrasound intensity in the range of 0.3-1.2 W / cm ² .

 As follows from the table. 4 and 5, it is enough to carry out the treatment for 10-20 minutes.

 In the table. 6 the results of two experiments are presented, justifying the drying parameters of the cultural fluid.

From the table. 6 shows that drying of the culture liquid at the temperature at both ²⁰ ° C and 40 ^{° C,} provides a formulation having the required catalytic activity.

Claims (3)

1. METHOD FOR PRODUCING A PLANT GROWTH STIMULANT, including cultivating an endophyte fungus, extracting a culture liquid with ethyl alcohol, filtering an alcohol solution containing a target product, characterized in that before extraction the culture liquid is treated with ultrasound with an intensity of 0.3 1.2 W / cm ² for 10 20 minutes and dried, and the dry residue of the culture fluid is subjected to extraction. 2. The method according to claim 1, characterized in that the culture fluid is dried at 20 40 ^o C.  3. The method according to claim 1, characterized in that they use an endophyte fungus isolated from the roots of sea buckthorn or half a floor.

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