RU2469538C1 - Agent for presowing treatment of seeds of pea - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: agent comprises salicylic acid, lipids from mycelium of fungus Ascichyta pinodes and the source of magnesium - magnesium sulfate MgSO₄ or magnesium chloride MgCl₂ at the following ratio of components: wt %: salicylic acid - 0.001, lipids from mycelium of fungus Ascichyta pinodes - 0.0001; the source of magnesium (magnesium sulfate MgSO₄; or magnesium chloride MgCl₂) - 0.001; water - 99.9979.

EFFECT: invention enables to increase the growth indices of pea, yield, disease resistance.

7 tbl, 7 ex

Description

The invention relates to agriculture and biotechnology, and in particular to means for presowing seed treatment, inducing disease resistance, resistance to pests and the yield of peas.

The rational use of environmental remedies in crop cultivation technologies does not provide for the complete extermination of pests and diseases, but their reduction to the threshold of harmfulness. At the same time, the crop is preserved, the harmful effect both on the plant itself and on the beneficial fauna and flora of the biocenosis is minimized. In this regard, the use of biological preparations and physiologically active substances has recently been considered a promising method of plant protection.

In recent years, metabolites of phytopathogenic fungi (elicitors) have been discovered, which are recognized by the plant and serve as a signal for triggering protective defense responses. Biogenic, i.e. isolated from microorganisms, elicitors cause the formation of protective substances in plant tissues. To date, about a hundred biogenic elicitors have been discovered, most of which are high molecular weight compounds of the lipoglycoprotein complex (LHPC).

Known is a presowing seed treatment drug NARCISS BP containing chitosan (50%), succinic acid (30%), glutamic acid (80%), which can be considered as a biological product (Handbook of pesticides and agrochemicals approved for use in the Russian Federation for 2004 year. The consumption rate of 1 l / t, it is recommended for use on rice, wheat, barley, sunflower, cucumbers. Publishing house "Agrorus", LLC "Agro 48", Lipetsk, p.318) [1].

The disadvantage of the drug described above is that it does not affect legumes, in particular peas.

The objective of the invention is to increase the disease resistance and productivity of pea seeds.

The problem is solved due to the fact that the known product containing organic acid (salicylic), according to the invention additionally contains lipids from the mycelium of the fungus Ascichyta pinodes and a source of magnesium (magnesium sulfate MgSO ₄ or magnesium chloride MgCl ₂ ) in the following ratio of components: wt.%:

Salicylic acid 0.001 Ascichyta pinodes mycelium lipids 0.0001 Source of magnesium (magnesium sulfate MgSO ₄ or magnesium chloride MgCl ₂ ) 0.001 Water 99,9979

When plants are treated with lipids of pathogenic fungi in plants, the phytoimmune potential increases, their tissue becomes able to respond more quickly and intensively to infection by pathogens, as a result of which the spread of infection is halved.

The method of lipid isolation from the mycelium of the fungus Ascichyta pinodes

The fungal mycelium is ground into a homogeneous mass using SiO ₂ . From a homogeneous mass, lipids are extracted with 70% alcohol (C ₂ H ₅ OH) in a ratio of 1 part of crushed mushroom and 9 parts of alcohol (1:10). Extracted by boiling in a water bath under reflux for 30 minutes. At the end of the extraction, the alcoholic extract of lipids and the insoluble sediment of the fungus are separated. Re-extraction was carried out in a water bath for 30 minutes with 70% alcohol (C ₂ H ₅ OH). Alcohol lipid extract and insoluble fungus precipitate are also shared. Combine both alcoholic lipid extracts. Alcohol was evaporated from the combined extract. Evaporation of alcohol is carried out in a water bath. After the cessation of boiling and bubbling, the flask stands in a water bath for 15 minutes (t water bath = 100 ° C). Then a sample is taken from the flask and a qualitative reaction is carried out for the presence of ethyl alcohol: 1 ml of the test solution containing alcohol is heated with 1 ml 1 n potassium hydroxide solution and a few drops of 0.1 n iodine solution to 50 ° C. In the presence of ethyl alcohol a yellow precipitate of iodoform - CHCl ₃ . Acetaldehyde and acetone give the same reaction. The reaction was negative, i.e. there was no alcohol. After that, distillation of the alcohol is stopped.

The resulting aqueous extract is cooled to n. (21-25 ° C), centrifuged at 4000 rpm for 15 minutes At the end of centrifugation, a precipitate of the lipoglycoprotein complex is obtained). Next, the Folch extraction of lipids is carried out with a mixture of CH ₃ OH-CHCl ₃ -H ₂ O in a ratio of 1: 2: 0.8, respectively. During extraction, intensive shaking is carried out for 1 hour. In this case, the system becomes transparent from white. The liquid exfoliates for 4 days. It turns out two phases: water and chloroform. The lower chloroform contains a lipid solution; the aqueous phase is not used further.

The solution of lipids in chloroform is evaporated at 30-40 ° C, lipids are quickly weighed and filled with phosphate buffer pH 7.8 (so that the oxidation of double bonds in unsaturated acids does not occur).

The value of magnesium in plants is determined by its occurrence in chlorophyll; participation in redox processes, the exchange of carbohydrates and organic acids; stimulating the formation of vitamin C and essential oils.

The resistance of plants to viruses can be increased by treating plants with salicylic acid. The effect of salicylic acid as a phytohormone is shown. Salicylic acid causes an increase in temperature in individual organs of thermogenic plants. The role of salicylic acid in the development of a non-specific reaction to stress factors and the accumulation of active oxygen species in cells is being actively studied.

The study of the effectiveness of the proposed tool and the biological activity of its components is carried out on test systems by measuring the activity of antioxidant enzymes: peroxidase and catalase, which is an express method for the accelerated evaluation of drugs (Method for assessing the resistance of peas to causative agents of fusariosis and ascochitosis according to biochemical parameters / Pavlovskaya N.E. ., Shalimova OA, Azarova EF Orel. - Orel GAU, 2002, 20 pp .; [2] Pavlovskaya N.E., Greenblatt A.I. Active forms of oxygen and apoptosis / Agricultural Biology, 2010 ) [3].

An indicator of the resistance of peas to pathogens can be an alternative oxidase system of cells: peroxidase and catalase, the activity of which can be used to judge the viability of plants. Peroxidase and catalase compete for the substrate - hydrogen peroxide and the activity of each enzyme separately depends on the work of the “partner”. As a rule, with an increase in the activity of the peroxidase enzyme, the activity of the catalase enzyme decreases.

Tests of the components of the proposed tool was carried out on pea seeds of varieties "Nord" and "Batrak".

Processing was carried out before germination for two hours.

Indications of the activity of catalase and peroxidase enzymes were measured during the first 10 days of germination, starting from the 3rd day.

Control options:

1 - option without processing, i.e. soaking pea seeds for 2 hours in water.

2 - Soaking for 2 hours in the industrial tool "Narcissus".

Test options (soaking for 2 hours):

1 - seeds treated with salicylic acid 0.001%;

2 - seeds treated with Mg 0.001%;

3 - seeds treated with lipoglycoprotein complex (LHPK) 0.0001%.

Example 1

The peroxidase enzyme activity was measured on pea seedlings on the 3rd, 5th, 7th, and 10th days of the experiment.

Table 1 Peroxidase activity in seedlings of peas of the Nord variety Processing options Peroxidase activity in cu every day 3rd day 5th day 7th day 10th day Control, seeds without treatment 69 96 101 122 Seeds treated with Narcissus 98 102 106 187 Seeds treated with salicylic acid 0.001% 102 113 145 166 Seeds Treated, Mg 0.001% 98 104 112 170 Seeds treated with LGA 0.0001% 96 102 123 169 Seeds processed by the proposed tool 133 149 198 212

It was revealed that in the control sample without treatment, the lowest activity of the peroxidase enzyme was observed, increasing during the experiment (69 cu, 96 cu, 101 cu, 122 cu).

In the control sample using Narcissus, the peroxidase enzyme activity indicators are higher and correspond to the following values: 3rd day 98 cu, 5th day - 102 cu, 7th day - 106 cu 10th day - 187 cu In the process of seedling growth, peroxidase activity is also steadily increasing.

When treating seeds with salicylic acid, it was found that the activity of peroxidase is higher in comparison with both controls and its activity also increases from the 3rd day to the 10th day (3rd day 102 cu, 5th day - 113 .e., the 7th day - 145 cu, the 10th day - 166 cu).

The use of magnesium showed even greater activity of the peroxidase enzyme with its increase as seedlings grew (3rd day 98 c.u., 5th day - 104 c.u., 7th day - 112 c.u., 10 day - 170 cu).

During the treatment of LGA pea seeds, the activity of the peroxidase enzyme also increases during the growth process (3rd day 96 cu, 5th day - 102 cu, 7th day - 123 cu, 10th day - 169 cu).

However, the highest peroxidase activity in the study was shown by pea seedlings, the seeds of which were treated with the proposed agent (3rd day - 133 cu, 5th day - 149 cu, 7th day - 198 cu, 10th day - 212 cu) (Table 1).

Thus, it was found that the activity of the peroxidase enzyme in all samples increases, however, the highest increase occurs in the sample treated with the proposed tool.

Example 2

Similar studies on the measurement of peroxidase enzyme activity were carried out on the pea variety “Batrak”.

table 2 The activity of peroxidase in seedlings of peas varieties "Batrak" Processing options Peroxidase activity in cu every day 3rd day 5th day 7th day 10th day Control, seeds without treatment 119 126 141 162 Seeds treated with Narcissus 138 152 166 230 Seeds treated with salicylic acid 0.001% 122 145 191 211 Seeds Treated, Mg 0.001% 120 167 182 188 Seeds treated with LGA 0.0001% 123 132 139 195 Seeds processed by the proposed tool 141 151 204 235

It was revealed that in the control variant without treatment, the activity of the peroxidase enzyme increased from 119 cu on the 3rd day to 162 cu by the 10th day.

In the control variant with the use of Narcissus, peroxidase activity increases from 138 cu from 3 days to 200 cu by the 10th day, which is higher in activity compared with the first control.

When studying the activity of peroxidase in variants treated with salicylic acid, a slight increase in activity was detected within the control variants: (3rd day - 122 cu, 5th day - 145 cu, 7th day - 191 .e., 10th day - 211 cu).

Higher activity indicators were found during the treatment of pea seeds with magnesium (3 days - 120 cu, 5 days - 167 cu, 7 days - 182 cu, 10 days - 188 cu).

Measurement of the activity of the peroxidase enzyme in pea seedlings under the influence of LGA revealed the following indicators: (3rd day 123 cu, 5th day - 123 cu, 7th day - 139 cu, 10th day - 195 c.u.) (Table 2).

Thus, we saw an identical picture of an increase in the activity of the peroxidase enzyme in the second grade of “Batrak” peas. Moreover, the highest rates are also found in the embodiment using the proposed tool.

Example 3

On the seedlings of the Nord pea variety, the activity of the catalase enzyme was studied in the same variants.

Table 3 Catalase activity in seedlings of peas of the Nord variety Processing options Catalase activity in cu every day 3rd day 5th day 7th day 10th day Control, seeds without treatment 3.9 3.3 3.0 1.1 Seeds treated with Narcissus 4.4 3.9 3,7 2,4 Seeds treated with salicylic acid 0.001% 5.3 4.9 4.1 2.6 Seeds Treated, Mg 0.001% 5.3 4.9 4.1 2.6 Seeds treated with LGA 0.0001% 6.1 4.6 4.0 2.2 Seeds processed by the proposed tool 5,6 4.3 4.0 2.0

In the control variant, without treatment, the activity of the catalase enzyme decreases (3rd day - 3.9 cu, 5th day - 3.3 cu, 7th day - 3.0 cu, 10 day - 1.1 cu).

In the control sample treated with Narcissus, the activity of catalase also decreases (day 3 - 4.4 cu, day 5 - 3.9 cu, day 7 - 3.7 .e., the 10th day - 2.4 cu).

When studying the activity of catalase in variants treated with salicylic acid, a decrease in the enzyme activity was also revealed: (3rd day - 5.3 cu, 5th day - 4.9 cu, 7th day - 4 , 1 cu, the 10th day - 2.6 cu).

When processing pea seeds with magnesium, it was revealed: (3rd day - 5.3 cu, 5th day - 4.9 cu, 7th day - 4.1 cu, 10th day - 2.6 cu).

Measurement of the activity of the catalase enzyme in pea seedlings under the influence of LGA revealed: (3rd day - 6.1 cu, 5th day - 4.6 cu, 7th day - 4.0 cu ., The 10th day - 2.2 cu).

The use of the proposed tool showed the following result (3rd day - 5.6 cu, 5th day - 4.3 cu, 7th day - 4.0 cu, 10th day - 2.0 cu) (Table 3).

Thus, the activity of the catalase enzyme in all test variants tends to decrease. However, in the control variant without treatment, such a decrease occurs more abruptly, which can lead to stressful consequences for the plant. The most smooth and painless decreases in all other options.

Example 4

Investigated the activity of the enzyme catalase and another variety of peas "Batrak".

Table 4 Catalase activity in seedlings of peas cultivar "Batrak" Processing options Day experiment 3rd day 5th day 7th day 10th day Control, seeds without treatment 4.0 2.9 3.0 2.5 Seeds treated with Narcissus 17,2 13,4 4,5 2,4 Seeds treated with salicylic acid 0.001% 4.6 3.2 2.1 1.7 Seeds Treated with Mg 0.001% 15.1 14.1 11.3 8.8 LGPK-treated seeds 12.1 11.1 9.5 7.6 Seeds processed by the proposed tool 10,4 9.3 7.8 6.4

In the control version, without treatment, the enzyme activity decreases (3rd day - 4.0 cu, 5th day - 2.9 cu, 7th day - 3.0 cu, 10- e day - 2.5 cu).

In the control sample treated with Narcissus, the activity of catalase also decreases (3 days - 17.2 cu, 5 days - 18.4 cu, 7 days - 4.5 .e., the 10th day - 2.4 cu).

When studying the activity of catalase in variants treated with salicylic acid, a decrease in the activity of the catalase enzyme was also revealed: (3rd day - 4.6 cu, 5th day - 3.2 cu, 7th day - 2.1 cu, the 10th day - 1.7 cu).

When treating pea seeds with magnesium, it was revealed: (3rd day - 15.1 cu, 5th day - 14.1 cu, 7th day - 11.3 cu, 10th day - 8.8 cu).

Measurement of the activity of the catalase enzyme in pea seedlings under the influence of LGPA revealed: (3rd day - 12.1 cu, 5th day - 11.1 cu, 7th day - 9.5 cu ., 10th day - 7.6 cu).

Processing by the proposed tool shows (3 days - 10.4 cu, 5 days - 9.3 cu, 7 days - 7.8 cu, 10 days - 6 , 4 c.u.) (Table 4).

Thus, a characteristic decrease in the activity of the enzyme catalase is observed, as is the case with the pea variety Nord.

Therefore, when studying the biological activity of the components of the proposed product on two medium-resistant varieties of peas "Nord" and "Batrak", a pattern was established of an increase in the activity of the peroxidase enzyme and a decrease in catalase activity. Pea seedlings quickly respond to treatment with biologically active substances by increasing or decreasing the activity of enzymes, which enhances peroxidase-dependent immunity in medium-resistant pea varieties.

Example 5

Based on the conducted laboratory experiments for field trials, a three-component proposed drug was made up of: salicylic acid 0.001%; Mg 0.001% and LHPK 0.0001%. Control options: 1 - control without processing, i.e. soaking before sowing in water; 2 control - processing was carried out by the industrial tool "Narcissus".

Table 5 The structure of the crop of peas varieties "Batrak". Treatment option Number of beans pcs. % to control Number of seeds pcs. % to control Seed weight g, from 1 plant % to control Mass 1000 seeds g % to control Control, seeds without treatment 2.09 - 3.81 - 14.65 - 203.99 - Seeds treated with Narcissus 1.92 -8.2 3.48 -8.7 12.84 -12.3 181.84 -15.4 Seeds processed by the proposed tool 2.16 +3.3 4.79 +25.0 17.59 +20.0 212.38 +1.3

It was revealed that when soaking seeds in the proposed tool, the number of beans on the plant increases to 2.16 pcs. on average, compared with the control without treatment (2.09 pcs.) and the control using Narcissus (1.92 pcs.), which is 3.3% higher compared with 1 control.

The number of seeds under the influence of the proposed funds is 4.79 pcs. on one plant on average, which is 25% higher than the control without treatment (3.81 pcs.) and the 2nd control (3.48 pcs.).

The weight of seeds on one plant with the use of the proposed tool is 17.59 g. This is a 20% increase in comparison with the control without treatment (14.65 g), and the weight of the seeds with the use of Narcissus is 12.84. The weight of 1000 seeds is the highest for the variant using the proposed product (212.38 g). (Table 5).

Thus, when analyzing the data obtained, it is seen that the number of beans on the plant, the number of seeds, the weight of seeds from one plant and the mass of 1000 seeds are the highest in the embodiment using the proposed means for soaking seeds.

Example 6

Table 6 Biological effectiveness of the use of physiologically active substances in the defeat of Nordic peas by diseases. Processing options The development of root rot,% (budding) The development of root rot,% (fruit formation) Rust,% (disease development) Ascochitosis,% (disease development) Control, seeds without treatment 42 71 58 28 Seeds treated with Narcissus 24 47 fifty 16 Seeds processed by the proposed tool 16 46 46 12

According to the results of the analysis of the biological effectiveness of presowing treatment of seeds, it was found that the development of root rot during budding under the influence of the proposed tool is reduced to 16%, while in the control sample without treatment the development of root rot is 42%, in the sample treated with Narcissus 24% By the period of fruit formation, the percentage of disease development increases in the sample treated with the proposed tool up to 46%, 1st control up to 71%, 2 and control up to 47%. Rust development is the lowest in the sample processed, the proposed tool and is 46%. Control 1 - 58%, control 2-50%. Regarding the development of ascochitosis, we observed a similar picture: control without treatment - 28%, treatment with Narcissus - 16%, treatment with the proposed tool - 12% (Table 6).

Thus, the results of the analyzes showed the greatest decrease in seed contamination with pathogenic and saprophytic microflora under the influence of the proposed tool for presowing treatment of seeds.

Example 7

To assess the role of biological products on the growth and development of pea plants most fully allows such an indicator as productivity.

Table 7 The productivity of peas varieties "Batrak" Processing options Productivity, t / ha % to control Control, seeds without treatment 5.62 - Control, seeds treated with Narcissus 5.96 +6.05 Seeds processed by the proposed tool 6.84 +22

When soaking pea seeds in the proposed tool, we found the highest yield indicators - 6.84 kg / ha, exceeding the control without treatment by 22% (5.62 kg / ha). When treating pea seeds with Narcissus, the yield is slightly lower than when processing with the proposed remedy - 5.96 kg / ha (Table 7).

Thus, it is shown that the highest yield of 6.84 kg / ha was found when using the proposed tool in the presowing treatment of pea seeds.

Therefore, the use of the studied biologically active substances contributes to an increase in peroxidase activity and a decrease in catalase activity, as indicators of increased resistance to non-specific pathogens, and also enhances growth processes at the seedling stage. When processing the proposed tool, the induction effect is more pronounced than in the control variants, which makes it possible to use it in the creation of an immunomodulating agent with a wide spectrum of action.

The mechanism of action of the proposed tool is that there is an activation of the synthesis of enzymes, in particular peroxidase, which lead to increased formation of phytoalexins, causing the formation of a whole complex of protective reactions.

Thus, the use of the proposed tool allows to increase growth indicators, productivity, resistance to disease.

Claims (1)

Means for presowing treatment of pea seeds containing organic acid, characterized in that it contains salicylic acid as an organic acid and additionally contains lipids from the fungus of the fungus Ascichyta pinodes and the source of magnesium is magnesium sulfate MgSO ₄ or magnesium chloride MgCl ₂ in the following ratio of components, wt. %:
Salicylic acid 0.001 Ascichyta pinodes mycelium lipids 0.0001 Source of magnesium (magnesium sulfate MgSO ₄ or magnesium chloride MgCl ₂ ) 0.001 Water 99,9979