RU2771962C1 - Method for activating the germination of radish seeds with hydrothermal nanosilicon under led lighting - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to the field of agriculture, in particular to crop production and biophotonics, and can be used in breeding in the selection of promising biotypes of plants responsive to artificial LED lighting. The method involves the treatment of seeds with hydrothermal nanosilicon using LEDs that generate low-intensity photons. Before sowing, radish seeds are pre-soaked for 2 hours in aqueous sol of hydrothermal nanosilicon, 0.005% concentration, followed by sowing on a substrate of mineral wool in the form of 20×20 cm plates at room temperature, 22-23°C, and moistening the seeds with water as the substrate dries. As light sources, monochromatic continuous illumination with LEDs of blue light with a wavelength of 440 nm, or green light with a wavelength of 525 nm, or red light with a wavelength of 660 nm is used when generating low-intensity photons in 6.52 Umol/(m²∙s), 1.44 Umol/(m²∙s) and 2.36 Umol/(m²∙s), respectively, at the level of the substrate with seeds for 6 days’ germination until the primary microgrowth is obtained.

EFFECT: method provides an expansion of the possibilities of using aqueous nanosilicon sols of hydrothermal origin in combination with exposure to a monochromatic spectrum of blue, green and red light regions of low intensities, constantly used in the process of germination of radish seeds with an increase in seed germination, the quality of sprouts in their productivity and the presence of high values of the total antioxidant activity of obtaining germinated seeds and primary microgreens of plants for nutrition and breeding in obtaining new highly productive biotypes of radish.

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Description

The invention relates to the field of agriculture, in particular to crop production and can be used in breeding when selecting promising plant genotypes responsive to artificial LED lighting and can be used for pre-sowing treatment of radish seeds to activate germination, increase, germination, quality of seedlings in terms of productivity in technologies obtaining germinated seeds and primary microgreens for a healthy diet.

In Russia, a state standard has been introduced for determining the germination of seeds of agricultural plants, which considers the conditions for germination of seeds and, as a rule, in the dark, taking into account temperature and time factors to assess the germination of seeds (GOST 12038-84. Seeds of agricultural crops. Methods for determining germination. - M. Standartinform, 2011). For seeds that are responsive to light, only natural light is considered in the standard.

Standards for germinating seeds under artificial light do not currently exist. For each plant, the issues of the influence of artificial lighting in its various components on the spectra of electromagnetic radiation, intensity and time of exposure at different stages of vegetation and photosynthesis are specifically studied in the development of technology elements for protected ground.

Close in technical solution to the proposed object is the use of presowing treatment of seeds with hydrothermal nanosilica of various concentrations for radish during germination in the dark to activate the germination process (RF patent No. 2747294, published 04.05.2021, Bull. No. 13). This method shows the effectiveness of the use of hydrothermal nanosilica (HSC) as an activation factor for the germination of radish seeds. However, the analogue does not take into account the influence of the lighting factor for activating the stage of radish seed germination with obtaining, in contrast to darkness, primary microgreens and germinated seeds under the influence of photons of the light flux generated by the LED emitter.

In the studies of many authors, the genetic specificity of the genome of different plants on the spectra of artificial illumination and the intensity of photon beams during photosynthesis of the formed apparatus of plant leaves is noted, not to mention the first stage of germination before the start of true photosynthesis, where the effect of the activation factor of nanoparticles with simultaneous exposure to photons of different intensities is poorly understood.

The closest technical solution to the proposed object is the solution set forth in the invention according to the patent of the Russian Federation No. 2746275 (published on April 12, 2021, Bull. No. 11), which shows a method for activating seed germination in relation to the agricultural root crop of sugar beet using seed treatment by soaking GNK concentration of 0.001% with constant 10-day LED illumination in the mode of generating low-intensity beams of monochromatic photons from LED lamps of blue light (LED SS) with a wavelength of 440 nm with an intensity of 6.52 μmol / (m ² ⋅ s), green light (LED GS ) with a wavelength of 525 nm with an intensity of 1.44 μmol / (m ² s). The impact of these and other concentrations of HNA on radish seeds during germination under such low-intensity monochromatic illumination by LED emitters is not known in practice.

There are no closer prototypes in terms of technical solutions for the effect of other modes of photon flux generated by LED emitters with significantly lower intensity indicators on radish seeds, starting from the primary phase of plant development.

EFFECT : expanding the possibilities of using hydrothermal nanosilica aqueous sols in combination with exposure to a monochromatic spectrum of low-intensity blue, green and red light areas used constantly in the process of germination of radish seeds after their treatment by soaking in a nanosilica solution of a certain concentration to increase seed germination, quality sprouts in terms of their productivity and the presence of high values of total antioxidant activity with the implementation of technologies for obtaining germinated seeds and primary plant microgreens for nutrition, for breeding when obtaining new highly productive radish biotypes.

The technical solution of the claimed object lies in the fact that the method includes the treatment of seeds with hydrothermal nanosilica using LEDs that generate low-intensity photons. Before sowing, radish seeds are pre-soaked for 2 hours in an aqueous ash of hydrothermal nanosilica with a concentration of 0.005%, followed by sowing on a mineral wool substrate in the form of 20x20 cm plates at room temperature 22-23 ° C and moistening the seeds with water as the substrate dries out using as light sources monochromatic continuous illumination LED blue light 440 nm, or green light 525 nm, or red light 660 nm with low intensity photon generation of 6.52 µmol/(m ² ⋅s), 1.44 µmol / (m ² ⋅s) and 2.36 µmol/ (m ² ⋅s), respectively, at the level of the substrate with seeds for 6 days of germination to obtain primary microgreens.

The method is carried out as follows.

The method uses an aqueous sol of hydrothermal nanosilica (HNK), which is obtained from natural hydrothermal solutions of the Mutnovsky deposit of Kamchatka (manufactured by Nanosilika LLC). In the tests, the initial HNK aqueous sol with a silica concentration of 2.5% was used. For seed treatment, the initial HNK sol was diluted with distilled water to a working concentration of 0.005% silica. Hydrothermal nanosilica has a high biochemical activity, a high rate of penetration into plant seeds, and a high sorption capacity for counting the size of silica particles and their surface area up to 500 cm ² /g. There are no toxic substances in the prepared working solution of hydrothermal silica, which gives the proposed solution a higher environmental friendliness and bioavailability for seeds, in particular, for endosperm and allows intensifying the process of seed germination not only in the dark to solve various biotechnological and breeding problems.

The size parameters of nanoparticles, predominantly in the range of 10-20 nm, are achieved by the capabilities of ultrafiltration equipment and technologies for the polycondensation of orthosilicic acid in hydrothermal solutions of the Mutnovsky deposit. This allows you to ensure high-quality processing of plant seeds.

As an object of research and verification of the method, radish of the Yubileiny variety was taken (Agrofirm Poisk LLC, Vereya village, Ramensky district, Moscow region).

Germination of radish seeds was carried out according to GOST 12038-84 with changes, namely: instead of filter paper, a mineral wool substrate in the form of plates 20*20 cm (400 cm ² ) was used. The number of seeds per 100 pcs. for each option, repeat three times. Irrigation was carried out with distilled water as the substrate dried. Germination was carried out at a temperature of 22-23°C. Germination of wheat seeds in the dark in accordance with GOST 12038-84 was used as a control, which were previously kept in distilled water for 2 hours before sowing, and in experimental variants, germination was carried out for 6 days using a monochromatic spectrum of light radiation with low photon intensity. For this, different versions of LED sources were used: blue light (LED SS) with a wavelength of 440 ns, green light (LED GS) with a wavelength of 525 nm, and red light (SS KS) with a wavelength of 660 nm in the mode of intensity of the generated photon flux of 6.52 µmol m ^-2 s ^-1 , 1.44 µmol m ^-2 s ^-1 and 2.36 µmol m ^-2 s ^-1 , respectively, at the level of the substrate with seeds during 6 days of germination.

Germination was determined in the experimental and control variants and the height of the sprouts and their weight were measured in 3 repetitions. The arithmetic mean was determined by germination and measured metric indicators of height and weight of 100 seedling samples.

To test the biological activity, the total antioxidant activity (CAOA) of the sprouts was determined. SAOA was determined by the coulometric method (in terms of rutin per 100 g of dry sample (d.d.) in accordance with MVI-01-00669068-13 (Zelenkov V.N., Lapin A.A. Total antioxidant activity. Measurement procedure on a coulometric analyzer. All-Russian Research Institute of Vegetable Growing. Vereya, Moscow region, 2013, 19c. Radish sprouts were dried in the shade at room temperature for 14 days. This method allows us to speak not only about antioxidant activity, but also about antiradical activity, since it is based on The method is based on the methodical approach of generation of bromine radicals with subsequent titration (using coulometry) of their solutions with aqueous extracts of germs of different variants of verification of the proposed method.

The results of testing the implementation of the method on the parameters of seed germination are shown in table 1-3.

Table 1. Germination (day 6, %) of seeds of radish cultivar Yubileiny for control options and in the proposed method

Experience variant Germination, % Change in germination in relation to control, % Germination of seeds in the dark - control 95 - Seed treatment with an aqueous solution of 0.005% HNK under illumination
LED SS (440 nm) and intensity
6.52 µmol m ^-2 s ^-1 95 0 Seed treatment with an aqueous solution of 0.005% HNK under illumination
LED AP (525 nm) and luminous flux intensity 1.44 µmol m ^-2 s ^-1 97 +2.1 Seed treatment with an aqueous solution of 0.005% HNK under illumination
SD COP (660 nm) and intensity
2.36 µmol m ^-2 s ^-1 97 +2.1

Table 2. Quality indicators of radish sprouts variety Yubileiny during germination for control options and the proposed method

Experience variant Sprouts height, cm Change in the height of sprouts relative to control, % Average weight of 100 sprouts, g Change in weight of 100 sprouts relative to control, % Germination of seeds in the dark - control 9.3 - 12.50 - Seed treatment with an aqueous solution of 0.005% HNK under illumination
LED SS (440 nm) and intensity
6.52 µmol m ^-2 s ^-1 8.8 -5.4 14.88 +19.0 Seed treatment with an aqueous solution of 0.005% HNK under illumination
SD AP (525 nm) and intensity
1.44 µmol m ^-2 s ^-1 9.2 -1.1 14.49 +15.9 Seed treatment with an aqueous solution of 0.005% HNK under illumination
SD COP (660 nm) and intensity
2.36 µmol m ^-2 s ^-1 7.5 -19.4 13.71 +9.7

As can be seen from tables 1 and 2, the use of nanosized silica has a positive effect on the stage of germination of radish seeds.

The use of GNK in pre-sowing seed treatment increases germination by 2.1% for options for using SD GS and SD KS in seed irradiation while maintaining the seed germination rate relative to the control for the option of irradiation with blue light 440 nm (Table 1).

For all tested variants, there is an increase in productivity during the germination of radish seeds by 9.7–19.0% by weight of 100 sprouts from the combined action of hydrothermal nanosilica and low-energy photons in the region of 440, 525, 660 nm (Table 2).

An essential point, both for germinated seeds and for primary microgreens and their use for nutrition, is the presence of primary photosynthesis pigments - chlorophyll for the experimental version of the proposed method compared with the control. In the control, germinated seeds are etiolated colorless sprouts, and in the experimental variant, the sprouts are green, which is typical for chlorophyll-containing plants with active photosynthesis.

Application of the proposed method for radishes allows you to get germinated seeds with an increase in productivity by sprouts (weight 100 sprouts, g) from 10.8% to 22.8%. At the same time, the height of radish sprouts in the tested variants of LED illumination with monochromatic light decreases from -1.1% (SD SS), -5.4% (SD SS) to -19.4% (SD KS), which confirms the prospects of purposeful obtaining of new biotypes of radish in breeding work on the search for highly productive plant varieties.

Table 3. Quality indicators of radish sprouts cv. Yubileiny according to CAOA (g rutin per 100 g w.o.) during germination for control options and the proposed method

Experience variant SAOA
in g rutin /100 g s.o. Change in CAOA compared to control, % Germination of seeds in the dark - control 17.03 - Seed treatment with an aqueous solution of 0.005% HNK under illumination with LED SS (440 nm) and intensity of 6.52 µmol m ^-2 s ^-1 17.23 +1.2 Seed treatment with an aqueous solution of 0.005% HNK under illumination with SD GL (525 nm) and intensity of 1.44 µmol m ^-2 s ^-1 17.72 +4.1 Seed treatment with an aqueous solution of 0.005% HNK under illumination with SD KS (660 nm) and intensity of 2.36 µmol m ^-2 s ^-1 17.89 +5.1

As can be seen from Table 3, the combined effect of hydrothermal nanosilica nanoparticles makes it possible to change the metabolome of sprouts in the direction of increasing their quality in terms of the content of metabolites that inhibit radicals with antioxidant activity, which is especially important in preventive nutrition using germinated seeds and microgreens.

The obtained experimental data allow us to conclude that nanosilica of hydrothermal origin is an activator of radish seed germination at the germination stage in combination with the use of low-intensity monochromatic illumination with blue, green or red light and can expand the scope of GNK in the technology of obtaining seed sprouts and further primary microgreens for a healthy diet , as well as in breeding work to create new varieties of radish.

Claims (1)

A method for activating the germination of radish seeds with hydrothermal nanosilica under LED lighting, including seed treatment with hydrothermal nanosilica using LEDs that generate low-intensity photons, characterized in that before sowing, radish seeds are pre-soaked for 2 hours in an aqueous ash of hydrothermal nanosilica with a concentration of 0.005%, followed by sowing on a substrate of mineral wool in the form of plates 20×20 cm at room temperature 22-23°C and wetting the seeds with water as the substrate dries using monochromatic continuous illumination of blue light LEDs with a wavelength of 440 nm, or green light with a wavelength of 525 as light sources nm, or red light with a wavelength of 660 nm when generating low-intensity photons of 6.52 µmol / (m ² ⋅s), 1.44 µmol / (m ² ⋅s) and 2.36 µmol / (m ² ⋅s) , respectively, at the level of the substrate with seeds for 6 days of germination until primary microgreens are obtained.