RU2307506C2 - Plant growth and development stimulator - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to preparation for stimulation of plant growth and development. Said preparation is prepared on the base of bacterium Halobacterium salinarum and contains bacteriorhodopsin in amount from 10^-9 M to 10^-6 M.

EFFECT: preparation effective in seed pretreatment and in plant cultivation in unfavorable climatic parameters.

3 tbl, 4 ex

Description

The invention relates to the field of crop production and agriculture and can be used to increase crop productivity and accelerate plant growth and development.

There are several ways to stimulate the growth and increase the productivity of plants, in particular by regulating the temperature and light exposure under artificial growing conditions in special structures. However, this requires significant costs for special equipment or devices, and the effect of stimulating growth and increasing productivity is not proportional to the labor expended.

In recent years, studies have been conducted on the effect of physiologically active substances isolated from plant tissues on the life of plants in the environment, that is, germination, growth, flowering, fruiting and aging.

However, so far only a limited number of such substances have been put into practical use, and only some of them are suitable for use in practice under existing conditions. Currently, 6 main groups of phytohormones are known: auxins, cytokinins, gibberellins, abscisic acid, ethylene and brassinolide

The most effective physiologically active substance and natural phytohormone bioregulator is brassinolide (epibrassinolide). According to its physiological effect, brassinolide monitors the balance of substances in the plant (homeostasis) and is an adaptogen, since it is involved in the synthesis of anti-stress proteins [1]. Currently, this bioregulator-phytohormone is being intensively studied [2-13].

In Russia, the registered trademark for the drug "EPIN-EXTRA R", made on the basis of epibrassinolide. "EPIN-EXTRA R" - a unique drug in the family of biostimulants, which in micromolar quantities provides:

- accelerated seed germination;

- Rooting seedlings during picking and transplanting;

- acceleration of maturation and increase in productivity;

- protection of plants from frost and other adverse conditions;

- increased resistance to late blight, perronosporosis, scab, bacteriosis and fusarium;

- the revival of weakened and rejuvenation of old plants due to the stimulation of lateral shoot formation;

- a decrease in the number of toxins, heavy metals, radionuclides in a plant,

- excess nitrates.

Especially shown is the use of "EPIN-EXTRA R" for:

- frost

- flooding,

- pest invasion

- processing plants before transplanting seedlings [1].

The disadvantage of this drug is:

- the use of complex organic solvents in its synthesis

- the danger of its use for human health and the environment

- low availability of the drug due to the high cost due to the complexity of its synthesis [13]

The task of the invention is to expand the arsenal of tools used as stimulants for the growth and development of plants.

The problem is solved by using Halobacterium salinarum as a stimulant of plant growth and development of plants.

The bacterial strains Halobacterium salinarum are natural inhabitants of salt lakes, where the concentration of NaCl can reach 4 M, which is 6 times higher than in sea water [14]. Bacteria of the genus Halobacterium salinarum do not have zoopathogenic or phytopathogenic properties and do not pose a danger to humans, therefore, working with them does not require special precautions [15].

The invention is illustrated by the following examples.

EXAMPLE 1. Obtaining a drug based on the biomass of bacteria Halobacterium salinarum.

The halophilic bacteria strain Halobacterium salinarum VKPM B-9025 is grown in 750 ml flasks in a volume of 300 ml of growth medium under fluorescent light illumination until the stationary stage of growth on a circular rocking chair at 37 ° C and shaking at 100 rpm. For cultivation, a medium of the following composition is used, wt.%: Peptone - 1, yeast extract - 0.5, NaCl - 25, MgSO ₄ - 2, KCl - 0.2, sodium citrate - 0.3, glycerin - 0.1, CaCl ₂ - 0.02, water - the rest, the pH of the medium is 7.2-7.4.

After 6 days, the precipitate was separated by centrifugation at 7000 g for 10 min and the resulting biomass was weighed. Then, distilled water in a volume of 300 ml was added to the precipitate (20 g) and incubated with stirring at room temperature for 16 hours. After centrifugation at 7000 g for 10 min, the resulting preparation is standardized by the content of bacteriorhodopsin in the supernatant, the main component of cell membranes, the amount of which is determined at a wavelength of 570 nm, and the concentration is calculated by the following formula:

C = D · M _r · V _p-pa · V _cuv / / E _br · V _samples

where D is the optical density of the solution at a wavelength of 570 nm;

M _r is the molecular weight of bacteriorhodopsin (26700);

V _p-pa is the total volume of the bacteriorhodopsin solution;

V _cuv is the volume of a solution of bacteriorhodopsin in a spectrophotometric cuvette;

E _br - the molar absorption coefficient of bacteriorhodopsin (63000 M ^-1 cm ^-1 );

V _sample - the sample volume of bacteriorhodopsin in a spectrophotometric cell.

The concentration of bacteriorhodopsin is 1 mg / ml.

EXAMPLE 2. The effect of the drug based on the biomass of bacteria Halobacterium salinarum on the germination energy and germination of oat seeds

The drug based on the biomass of bacteria Halobacterium salinarum (hereinafter HS) is used as a stimulator of plant growth and development in the experiment with seedlings. The following served as controls: control 1 - water and control 2 - solution of the EPIN-EXTRA R preparation (hereinafter referred to as Epin) (lot No. 1, TU 2387-002-42719567-98, state registration number: 09-02901-0082- 1, the shelf life of the drug is until March 2007, the manufacturer is NNPP "NEST M") [16].

The experiment is carried out in accordance with the standard method [17-21], when the indicators of the intensity of the formation of plant roots reflect the germination energy and germination of seeds. These indicators were recorded, respectively, on the 5th and 10th days after the start of the experiment. As a test culture, oats of the Comet variety are used. The seeds of this culture are soaked for 1 day in a solution of HS or Epin or water. After a day, the seeds are laid for germination. To study the effectiveness of the drug HS and Epin, the following concentrations of samples are taken: Epin according to the recommended instructions at a concentration of 0.5 × 10 ^-7 M, and HS - 0.4 × 10 ^-7 M per bacteriorhodopsin. The experiment is laid in triplicate in Petri dishes. Each variant contains 60 pieces of seeds (20 pieces of seeds in a cup). To soak 60 pieces of seeds spend 5 ml of a solution of the indicated concentrations or water. The results are shown in table 1.

Table 1 Option The energy of seed germination. Number of sprouted seeds (%) 5 days 10 days Control 1 63 63 Control 2 92 93 HS drug 90 92

On the 5th and 10th day after the oat seed treatment in the claimed HS preparation, the seed germination energy is the same as in control 2 (Epin) and 1.5 times higher than when soaked in water (92% versus 63%, respectively). The HS preparation contains only water, while Epin contains 0.05% alcohol, in addition, working with the HS drug, unlike the Epin preparation, does not require individual protective equipment.

Other cereal growth stimulants are less effective. Thus, the complex organic substance decarbamide nickel iodide increases seed germination by 14–40% compared to control [22], and BTTM (a mixture of tetramethylurea with boric acid) increases laboratory germination of spring wheat seeds by 14% [23]. The stimulation of acenaphthene is also lower than that of the HS preparation, the percentage of germination of millet and barley seeds (62% and 27%, respectively) on day 7 [24].

EXAMPLE 3. The effectiveness of the drug based on the biomass of bacteria Halobacterium salinarum in the pre-sowing treatment of oats and rape seeds.

To ensure high accuracy of the results of vegetation experiments, they carry out all the necessary work on the special preparation of soil, vessels and nutrient solutions for growing plants [5-9].

The soil (arable horizon) is selected in the field, then slightly dried and sieved through a sieve with a mesh size of 3 mm. After thorough mixing in the soil, moisture is determined to calculate the irrigation weight of the vessels. The vessels selected by weight are rinsed with tap and then distilled water. To ensure normal conditions for the development of the root system of plants, drainage is laid at the bottom of the vessel (broken glass covered with gauze); a glass aeration tube is placed on the glass through a hole in the gauze, which is carefully pressed against the vessel walls with wet sand. Taring vessels to the same weight is carried out with a small amount of sand. Before setting up the experiment, test stuffing is carried out to determine the optimal soil level in height (2-2.5 cm from the upper edge of the vessel).

The agrochemical characteristics of the soil are as follows: humus content - 0.98%; P ₂ O ₅ - 7.7 mg / 100 g; K ₂ O - 1.8 mg / 100 g; pH _kcl - 4.0; Ng. - 2.5 mEq / 100 g, that is, the soil taken for the experiment is highly acidic, poor in nutrients. On this type of soil, one can observe not only the growth-promoting properties of the claimed HS preparation and Epin preparation (control 2), but also their adaptogenic properties (growth ability on soil depleted in nutrients). The experiment is laid in triplicate in vessels containing 3.6 kg of prepared soil. The soil moisture in the vessels is maintained at 60% of the total field moisture capacity by daily watering of the vessels by weight. As sources of nitrogen, phosphorus and potassium, 0.1 g of nutritional salt is added per vessel (nutritional salts: nitrogen — NH ₄ NO ₃ ; phosphorus — Ca (H ₂ PO ₄ ) ₂ · N ₂ O and potassium — K ₂ SO ₄ )

As test cultures, oats of the Comet variety and spring rape of Lugovskaya are used. 60 pieces of seeds of each of these plants are treated with 5 ml of solutions of the drug HS, control 1 and control 2, as in example 2. The next day after treatment, the seeds of oats and rapeseed are sown. The repetition of the experience of each option is threefold. Harvesting oats and rapeseed for green mass is carried out on the 40th day after sowing. Harvested plants are weighed, then dried to an air-dry state. The results are presented in table 2.

table 2 Option Oats Rape Height (cm) Weight Weight (g / vessel) Add to dry weight control Height (cm) Weight Weight (g / vessel) Add to dry weight control raw dry g / vessel % raw dry g / vessel % Control 1 58 65 10.1 - - 45 48 5,4 - - Control 2 57 61 10.3 0.2 2.0 51 53 6.3 0.9 16.7 HS drug 60 66 10,4 0.3 3.0 fifty 57 6.7 1.3 24.1 NDS ₀₅ 0.45 g / vessel 0.70 g / vessel

In the growing experiment, the growth-stimulating effect of Epin and HS preparations was statistically significant with the smallest statistical difference (NDS ₀₅ ) in oats 0.45 g / vessel and rape 0.70 g / vessel. The growth-promoting and adaptagenic effect of the claimed HS preparation on plants is 1.5 times higher than that of the Epin preparation, while maintaining the other advantages of HS indicated above. For comparison, the use of Epin in combination with jasmonic acid gives a significantly lower increase in wheat mass compared to Epin (20%) [25] than the claimed HS preparation (50%) for oats and canola.

EXAMPLE 4. The effectiveness of the drug based on the biomass of bacteria Halobacterium salinarum when it is applied to the soil used for sowing oats and canola.

All necessary work, consisting in the special preparation of soil, blood vessels and nutrient solutions for growing plants, is carried out as in example 3. The inventive preparation HS is introduced into the soil of vessels under rape and oats in a solution of 5 ml and a concentration of 0.4 × 10 ^-6 M per the vessel, and the Epin preparation is used according to the instructions, namely: the seeds of oats and rapeseed are treated with Epin solution and the seeds are sown as in example 3 (control 2). The seeds of oats and rapeseed are treated with a solution of distilled water and the seeds are sown in example 3 (control 1).

Harvesting oats and rapeseed for green mass and analysis of harvested plants is carried out as in example 3. The results are presented in table 3.

Table 3 Option Oats Pane Height (cm) Weight Weight (g / vessel) Add to dry weight control Height (cm) Weight Weight (g / vessel) Add to dry weight control raw dry g / vessel % raw dry g / vessel % Control 1 58 65 10.1 - - 45 48 5,4 - - Control 2 57 61 10.3 0.2 2.0 51 53 6.3 0.9 16.7 HS drug 62 65 10.9 0.8 8.0 52 59 6.7 1.3 24.1 NDS ₀₅ 0.45 g / vessel 0.70 g / vessel

 The results presented in table 3 are statistically significant. They confirm that the growth-promoting and adaptagenic effect of the claimed drug HS, introduced into the soil, is preserved. It is higher than that of the Epin preparation, 1.5 times on rapeseed, and on oats - 4 times.

Thus, the inventive HS preparation has growth-promoting and adaptogenic properties that are not inferior, and in some cases superior to those of the currently widely used growth stimulator of the Epin preparation.

A significant advantage of the claimed drug is its environmental friendliness, since it is non-toxic, soluble in water and obtained by microbiological synthesis based on non-pathogenic bacteria. The drug is effective in micromolar quantities both during pre-sowing treatment of seeds and when it is applied to the soil before sowing. HS can be recommended for processing soils and seeds when growing crops in adverse climatic conditions.

Information sources

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13. http://www.vitusltd.ru/ch-epin-br.html.

14. Cavalier-Smith T. The neomuran origin of archaebacteria, the negibacterial root of the universal tree and bacterial megaclassification. Int. J. Of Syst. and Envir. Microb. 2002, 52, 7-76.

15. Sanitary rules of the joint venture 1.2.731-99.

16. http://senpolia.tskm.ru/preparats/epin.shtml.

17. A.V. Sokolov. The technique of field and vegetation experiments with fertilizers and herbicides. - M .: Nauka, 1967, 180 p.

18. 3.I. Zhurbitsky. Theory and practice of the vegetative method. USSR Academy of Sciences. Publishing House "Science". Moscow, 1968, 244 p.

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21. Methodological recommendations for laboratory screening of synthetic plant growth stimulants. Sep. NIITEkhima. Cherkasy, 1985 .-- 1-28 s.

22. A.S. RF N 1639452.

23. RF patent N 2179806.

24. RF patent N 2088084.

25. RF patent N 2145165.

Claims (1)

A preparation for stimulating growth and development based on bacteria Halobacterium salinarum VKPM B-9025, containing bacteriorhodopsin in an amount of 10 ^-9 to 10 ^-6 M.