RU2386252C2 - Means for improving systemic induced persistence of feed crops to, for instance fungal pathogenic agents - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: product for improving systemic induced persistence of feed crops to fungal pathogenic agents contains 2·10^-8 M of salicylic acid, extracts of medicinal herbs such as purple echinacea and Greek-valerian polemonium, 2·10^-8 M of lectins obtained from kidney beans, 0.2 M of magnesium sulfate and 0.2 M of zinc sulfate with equal ratio of the components by weight.

EFFECT: use of the suggested product will render unnecessary the practice of repeated presowing treatment of seeds, will combine use of the product with a technique of presowing treatment of seeds using ordinary disinfectants as well as improve systemic induced persistence to fungal pathogenic agents.

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Description

The invention relates to agriculture and biotechnology, in particular to means for presowing treatment of seeds that increase systemic induced resistance (SIU).

A wide range of growth substances and plant protection products is known from weeds, pests and diseases (1. Malamy J., Carr JP, Klessig DF, Raskin I. Salicilik acid: A Likely Endogenous Signal in the resistane response of tobacco to Viral Infection // Science 1990. V.250. P.1002-1004; 2. Raskin I. Salicylate, a new plant hormone. // Plant Physiol. - 1992. - 99, No. 3, p. 799-803).

The disadvantages of these funds include the use of chemical compounds that pollute the environment and pose a danger to human and animal health, the great complexity of their manufacture.

The closest analogue to the proposed tool is a tool containing salicylic acid and having a growth-promoting effect (NNPP "NEST M" drug Zircon (0.1 g / l mixture of hydroxycinnamic acids) // Patent RU 2257059 (prototype).

Pre-sowing seed treatment with salicylic acid at a concentration of 0.2 mM helps to accelerate their germination, growth processes in young plants, and also leads to a noticeable increase in yield, which indicates the prospects of its use as a growth regulator (Sakhabutdinova AR [et al.] The effect of seed treatment with salicylic acid and fungicide phenoram super on the hormonal status and growth of wheat seedlings // Agrochemistry, 2004, No. 4, p.17-21).

Salicylic acid with a concentration of less than 0.2 mM plays a key role in the induction of systemic acquired resistance to phytopathogenic fungi, bacteria, viruses, and at the same time increases non-specific resistance of plants (Ozeretskovskaya OL Induction of plant resistance // Agrarian Russia. - 1999. - No. 1. - p. 4-9).

The objective of the invention is a single pre-sowing treatment, full compatibility with the technology of pre-sowing seed treatment with traditional dressers, as well as increasing the systemic induced resistance of feed crops to fungal pathogens.

The problem is solved due to the fact that in the known preparation for increasing the systemic induced resistance of feed crops to fungal pathogens containing 2 · 10 ^-8 M salicylic acid, according to the invention, extracts from medicinal plants Echinacea purpurea and Cyanosis blue, 2 · 10 ^{-8 are} additionally used M lectins from bean seeds, 0.2 M magnesium sulfate and 0.2 M zinc sulfate in the following ratio of components: Echinacea purpurea extract: Cyanosis blue extract: bean seed lectins: salicylic acid: MgSO ₄ : ZnSO ₄ = 1: 1: one : 1: 1: 1.

At least four polypeptide hormones, lectins, PR proteins, salicylic and jasmonic acids, as well as some other compounds, are involved in the induction and coordination of protective antiviral reactions of plants with SIU. It is known that the development of pathogenesis in plants is accompanied by the formation of peroxide radicals that cause destructive processes in the structure of cell membranes (Methods of biochemical study of plants. Under the editorship of Ermakov AI - M .: Agropromizdat, - 1987. 431 p.).

In this regard, the idea of the therapeutic use of bioimmune molecules to suppress peroxidation reactions is of particular importance. Thus, these elicitor molecules are potentially capable of acting as inducers of the systemic resistance of plants.

However, environmental requirements lead to the need to create drugs that belong to the category of biological products, the use of which would be effective in small doses.

Preparation of raw materials includes the processes of weighing powders, measuring liquids, grinding plants.

To prepare the extracts of Echinacea and Sinyukha, low-mineralized spring water was used that meets the requirements of CaN PiN 2.3.2.1078-01 from the natural source of the Oryol Polesie National Reserve. Spring water is subjected to sterilization before being fed to the extraction. For this, the water is poured into a glass container, corked and steam sterilized in an autoclave for 30 minutes at a pressure of 1.5 atm and a temperature of 119-120 ° C.

Medicinal plants for the production of extracts are taken in the following ratio: 1 part of Echinacea and 1 part of Cyanosis. The dried parts of the plant (roots and ground parts in the flowering stage) are crushed in a laboratory mill. The size of the crushed particles of medicinal herbs is 3-5 mm. Next, extraction is carried out with a 40% aqueous-alcoholic solution according to the standard procedure for 10-12 hours at room temperature. The output of the phytocomposition is 87.69%.

Next, pectins from bean seeds are isolated and purified as follows:

Extraction of crushed seeds 0.9 M NACl 1 hour (100 g flour: 900 ml NaCl). ↓ Centrifugation (2000g 20 minutes). ↓ Extraction and centrifugation (repeatedly) in the same mode of 0.9 M NaCl 500 ml. The combined supernatants were adjusted to pH 4.0 with 4 n HCl. ↓ Centrifugation (2000g, 20 minutes). The supernatant is neutralized to pH 7.0 ↓ The precipitate is removed by centrifugation (2000g, 20 minutes). ↓ Protein is salted out with 60% ammonium sulfate. ↓ The crude (crude) lectin is collected on a filter and dissolved in five times (by volume) the amount of distilled water. ↓ Dialysis: 24 hours at 110 ° C against a buffer mixture (0.1 M acetate buffer, pH 6.8) ↓ The lectin solution is clarified by centrifugation (6000g, 20 minutes). ↓ The clarified lectin is applied to a column of Sephodex G-75, balanced with a buffer mixture. ↓ Adsorbed lectin is eluted with a 0.1 M glucose solution in 0.1 M acetate buffer (pH 6.8). The protein is determined spectrophotometrically. ↓ The eluate protein is precipitated with 70% ammonium sulfate. ↓ The precipitate was separated by centrifugation (2000g, 30 minutes) and dissolved in water. ↓ Dialysis: 18 h at 14 ° C against 0.1 M acetate buffer (pH 6.8). ↓ The precipitate formed is removed by centrifugation (6000g, 30 minutes). ↓ Dialysis: 24 hours at 14 ° C against distilled water. ↓ The formed lectin crystals are dissolved in water (acidified with 0.1 N HCl to pH 4.0), the solution is neutralized to pH 6.7-7.0 and freeze-dried.

As a result, a lectin preparation with a hemagglutinin activity of 27 (mg / ml) ^{-1 was} obtained.

Then, salicylic acid, magnesium and zinc sulfates are weighed in the required amount on a weight batcher and fed to the stage of solution preparation.

Then, pre-prepared dissolved components are mixed (extracts of Echinacea and Sinyukha, bean pectins, salicylic acid, magnesium and zinc sulfates) in a ratio of 1: 1: 1: 1: 1: 1.

Further, to prepare 1 liter of working solution, the prepared dissolved components are diluted in a proportion of 20 ml (tablespoon of the drug) per 1 liter of water. Then this solution is mixed with a 2% aqueous solution of CMC-sodium (wallpaper glue), which is a traditional seed incrustator in a ratio of 1: 1 by volume.

The effectiveness of this tool was tested for biological activity, carried out by measuring the value of peroxidase activity, which is the express method of accelerated drug evaluation (Tarchevsky I.A. Elixitor-induced signaling systems and their interaction. // Plant Physiology, 2000, No. 2, p.321 -331. Method for assessing the resistance of peas to the causative agents of fusarium and ascochitosis according to biochemical parameters / Pavlovskaya N.E., Shalimova O.A., Azarova E.F., Orel. - Orel State Agrarian University, 2002, 20 pp.).

In the experiments, 1500 pieces of pea seeds of the Orlus variety were used. 3 groups of 500 seeds were formed.

Example 1. The first group of seeds (control) was treated with a traditional CMC-sodium incrustator. Seeds were placed in a container and mixed with an incrustator on a rocking chair for 15 minutes. Then the seeds were placed in the plant on a filter paper moistened with water. Studied the energy of seed germination (table 1). In order to study the systemic induced resistance, three-day-old pea seedlings were treated with Fusarium oxyspomm pathogen fungal culture fluid and peroxidase activity in infected seedlings was studied (Table 2).

Example 2. They took 500 pieces of pea seeds, placed in a container and mixed with the powdered preparation "ECOST" on a rocking chair for 15 minutes. Seeds were placed in the plant on a filter paper moistened with water. The results of germination energy are presented in table 1. In order to study the systemic induced resistance, three-day-old pea seedlings were treated with Fusarium oxysporum pathogen fungal culture fluid and peroxidase activity in infected seedlings was studied (table 2).

Example 3. They took 500 pieces of pea seeds, placed in a container and mixed with the proposed tool on a rocking chair for 15 minutes Seeds were placed in the plant on a filter paper moistened with water. Studied the energy of seed germination (table 1). In order to study the systemic induced resistance, three-day-old pea seedlings were treated with the culture fluid of the fungus pathogen Fusarium oxysporum and peroxidase activity in infected seedlings was studied (table 2).

Table 1 The effect of presowing seed treatment on the growth energy of peas of the Orlus variety Experience group Number of seeds Germination energy,% Germination,% Number of beans per 1 plant, pcs. 1. Control 500 86.6 84.8 1.95 2. ECOST 500 87.5 91.3 2.12 3. The proposed tool 500 88.0 84.0 2,32

Thus, it is seen that the use of the proposed tool will increase the energy of growth of legumes.

table 2 Peroxidase activity in pea seedlings when processing infected plants, in arbitrary units of activity. Days The control The drug "ECOST" (prototype) Suggested remedy four 201 330 430 5 219.5 333.3 454.5 6 2250 652.2 535.7 7 1875 1666.7 1875 8 1153.8 1875 2142.9

As a result, it was found that pre-sowing treatment with the proposed biological product increases the energy of germination of peas, contributes to an increase in peroxidase activity. Moreover, the drug causes a more intense activation of peroxidase in comparison with the industrial drug "ECOST".

One of the natural mechanisms of plant resistance to fungal infection is a hypersensitivity reaction, which can be judged by the development of necrosis. So, on the lower inoculated pea leaves, pre-treated with the proposed tool, the number of recorded necrosis was significantly less in comparison with the control. At the same time, in the upper uninfected leaves, necrosis was not recorded at all (table 3).

Table 3 The dynamics of the formation of necrotic spots on infected pathogenic Fusarium oxysporum pea leaves after seed treatment Experience group Day after infection one 2 3 four 5 6 7 8 9 10 eleven 12 1. Control - - - - - - + + + + + + 2. ECOST - - - - - - - - - + + + 3. The proposed tool - - - - - - - - - - - + (+) - the presence of necrotic spots was established; (-) - absence of necrotic spots.

This indicates an increase in systemic induced resistance of feed crops to fungal pathogens under the influence of the proposed tool.

Thus, the use of the proposed tool will eliminate the need for multiple presowing treatment of seeds, combine the use of the tool with the technology of presowing treatment of seeds with traditional dressers, as well as increase the systemic induced resistance to fungal pathogens.

Claims (1)

Means for increasing the systemic induced resistance of forage crops, for example peas, to fungal pathogens, including 2 · 10 ^-8 M salicylic acid, characterized in that it additionally contains extracts from medicinal plants Echinacea purpurea and Cyanosis blue, 2 · 10 ^-8 M lectins beans, 0.2 M magnesium sulfate, 0.2 M zinc sulfate with a ratio of components by weight Echinacea purpurea extract: Cyanosis blue extract: bean seed lectins: salicylic acid: MgSO ₄ : ZnSO ₄ = 1: 1: 1: 1: 1: 1.