RU2366153C1 - Method of berry and decorative bush plant nursing in vitro in non-sterile conditions - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention concerns agriculture, particularly nursing gardens. Method involves microplant replanting from test tubes into cartridges filled with artificial substrate. Dehydrated stabilised sediments of city sewage and neutralised high-moor peat taken at the ratio of 1:4 are used as substrate. Replanting is performed in the beginning of January. Microplants replanted into cartridges are nursed under sealed film cover for two weeks.

EFFECT: earlier replanting of microplants into non-sterile environment, enhanced survivability, growth and development of regenerated plants, improved planting material quality for berry and decorative bushes.

2 cl, 9 tbl, 7 ex

Description

The invention relates to the field of agriculture, namely to nursery, and can be used in the technology of microclonal propagation of berry and ornamental shrubs.

Known methods of adaptation and growing microplants in conditions of high humidity on artificial substrates consisting of an organic base (coniferous bark, peat, compost, flax fire, soil, sawdust) (Pocock S. Procedures and problems associated with the transfer of tissue- cultured plants / Comb. Proc./ Intern. Plant Propagators Soc, 1984; T. 33, p. 316-320; Orlikowska T. Rozmnazanie pigwy S 1 w kulturach in vitro / Ogrodnictwo, 1988; T. 25. N 4, - s.6-8; Tsvetkov I., Krstanova S. Adaptation to chrysanthemum, obtained in different species and substrates / Plant Science, 1991; Vol. 28. N 3-6, - p. 44-49) and inert materials (sand vermiculi t, mineral wool, perlite, zeolite) (Dzheneev S.Yu., Litvak AI, Soldatov BC, Nasimov AZ Use of artificial ionic soil BIONA for adaptation of grape plants cloned in vitro. 1990, - 14 p. Manuscript of the Depot at VNIITEIagroprom October 31, 1990; Yalta; 1990. - Bibliography 23; Youn Y., Ishil K., Saito A., Ohba K. In vitro plantlet regeneration from axillary buds of mature trees of Tilia cordata J. Japan. Forestry Soc, 1988; T. 70. N 7, p.315-317; Wainwright H., Scrace J. Influence of in vitro preconditioning with carbohydrates during the rooting of microcuttings on in vivo establishment / Sc. hortic, 1989; T.38. N 3/4, p. 261-267; Eduardo da Costa Nunes, Carolina Volkmer de Castilho, Fabio Netto Moreno and Ana Maria Viana. In vitro culture of Cedrela fissilis Vellozo (Meliaceae), Plant Cell, Tissue and Organ Culture, 2002, V.70, N.3).

Typically, such substrates require disinfection or treatment with hot steam. To avoid this, tube plants are adapted in inert materials, such as zeolite, vermiculite, coarse perlite, and mineral wool (Batukaev A.A., Sadaeva M.A. Use of zeolite for adaptation and growing of grape plants in vitro in vivo. intensification and cooperation in the selection of horticultural crops and grapes. - Krasnodar, 2002, p. 212-215; Einset JW, Alexander JH Multiplication of Syringa species and cultivars in tissue culture / Comb. Proc. / Intern. Plant Propagators Soc, 1985; T. 34, p.628-636; RLM Pierik, HHM Steagmans, PFSprenkels Micropropagation of Lilac (Syringa vulgaris L.) Biotechnology and Forestry, 1992, V. 20, H. 407-426). Such artificial substrates do not need to be sterilized, but they do not contain the nutrients necessary for the growth and development of plants.

In the technology of clonal micropropagation, a mixture of perlite and lowland peat is most often used, which is decontaminated with a 0.1% solution of benlat, euparen (Mikhalchik L.S., Demenko V.I. Reproduction of apple and cherry in vitro / Scientific Research Conference of young scientists 14- June 17, 1988, 1988, p.649-657 Manuscript of the Depot at VNIITEIagroprom 05.01 1989), antimicrobial solutions with the addition of terrazole (Pocock S. Procedures and problems associated with the transfer of tissue-cultured plants / Comb. Proc./Intern Plant Propagators Soc, 1984; T.33, p. 316-320) or treated with hot steam (Orlikowska T. Propagation of quince S 1 (Cydonia blonga Mill.) In vitro Fruit Sc. Rep. Skierniewice, 1988; Vol. 15 N 4, p. 157-165).

The closest to the invention in terms of essential features is a method for adapting regenerants of fruit and ornamental crops in non-sterile conditions on substrates containing lowland peat and perlite in equal proportions (Leontiev-Orlov O.A. Production of apple planting material by tissue culture method. Moscow, 1986, 24 pp .; Mirzaev MM Effective methods of growing clove meristemic plants in soil substrates, Trudy Shreder Research Institute of Horticulture, Viticulture and Winemaking, 1988; T.50, p.31-34; Catherine A. Offord and Lindsay C. Campb ell Micropropagation of Telopea speciosissima R. Br. (Proteaceae). Rhizogenesis and acclimatization to ex vitro conditions, Plant Cell Tissue and Organ Culture, 1992, V.29, Number 3). According to this method, in March-April, microplants of garden crops are transferred to heated substrates on artificial substrates and placed for 4-5 weeks in closed plastic boxes or under film shelters where a high relative humidity of 65-80% and a temperature of 25-28 ° C are maintained after which the plants acclimatize in a greenhouse when exposed for 16 hours from March to early May (Einset JW, Alexander JH Multiplication of Syringa species and cultivars in tissue culture / Comb. Proc. / Intern. Plant Propagators Soc, 1985; T. 34, - p. 628-636; Refouvelet E., Le Nours S., talon C., Daguin F. A new method for in vitro propagation of lilac: reg rowth and storage conditions for axillary buds encapsulated in alginate beads, development of a pre - acclimatization stage. Scientia Horticulturae, 1998, 74, 233-241).

The disadvantages of this method are: the need for steaming or disinfecting the substrate; late terms of transplantation for acclimatization and growing, large lunges (up to 30-40%) in vitro of plants in non-sterile conditions (waterlogging, insufficient nutrient content), slow initial growth and slow growth after transplanting plants from cassettes into containers, poor development of the aerial part and the root system, which does not make it possible to grow planting material with closed roots by the beginning of the growing season - spring sales or the laying of perennial plantings.

The claimed invention is directed to an earlier (first half of January) transplantation of microplants into non-sterile conditions for adaptation and growing, increasing survival, enhancing the growth and development of regenerants in ex vitro conditions, increasing the yield of viable high-quality planting material of berry and ornamental shrubs with a closed root system to the beginning vegetation period due to favorable physicochemical, aseptic properties and high hormonal activity of the substrate used.

To solve this problem, explants of berry plants and lilacs are introduced into a sterile culture in March, in the first half of January microplants are transplanted from test tubes into plastic cassettes with small cells (d 4 cm, V - 10 cm ³ ) on artificial substrates consisting of stabilized dehydrated precipitation of urban wastewater (OGSV), not more than one year of storage and neutralized (pH 6-6.5) peat in a ratio of 1: 4. The substrates are not pre-disinfected or steamed, they are prepared by simple mixing of the components. The first two weeks after the transplantation, the microplants are kept in cassettes under airtight film shelters on racks with an expanded claydite layer (2-3 cm) at a temperature of 22-26 ° С and additional exposure from the beginning of January to the end of February (illumination 2000 lux, 16 hours). In early April, microplants with an intact root lump are transplanted from cells into containers of 1.6 l (d 11 cm).

The proposed method of growing plants in vitro on substrates containing OGSV and neutralized peat, has several advantages over the prototype:

- 2.5 months earlier, test plants are transplanted into non-sterile conditions for adaptation and growing;

- growing on a substrate with OGSV provides high (up to 95-100%) survival and preservation in vitro of plants in non-sterile conditions;

- microplants quickly move to growth and are characterized by high growth and development rates;

- a significant leaf surface and growth are formed, which, along with a more powerful development of the root system (increasing the number and mass of roots, their total length) makes it possible to transplant in vitro plants with a closed root system to a permanent place without growing in the nursery;

- the total yield of viable in vitro plants increases by a factor of 1.5-1.8, the period from the introduction of explants into a sterile culture to the transplantation of plants to a permanent place is reduced, the efficiency of using expensive protected soil is increased;

- due to the aseptic properties of high peat and the absence of pathogenic microflora in fresh OGSV there is no need for preliminary disinfection or steaming of the substrate, which saves the cost of manual labor, money and prevents environmental pollution;

- the use of OGSV as a part of substrates for adaptation and growing microplants is one of the methods for their utilization.

The essential features characterizing the invention are transplant of regenerants into non-sterile conditions in the first half of January, the use of an artificial substrate containing stabilized sewage sludge (OHSW), not more than a year of storage and neutralized peat in a ratio of 1: 4 without prior disinfection or steaming.

Causal relationships are that dehydrated stabilized sludge from urban wastewater (OHSW), obtained by mechanical treatment and thermophilic anaerobic digestion, in which more than 30 systematic groups of microorganisms take part, are an organomineral suspension, the organic part of which consists of carbohydrates, proteins (bacterial mass), lipids, lignin, humic substances, wax and chitin.

Fermented mechanically dehydrated sludge contains a high amount of basic macro- and microelements and is a valuable organic fertilizer equivalent to manure and sapropel. The precipitate obtained at Moscow aeration stations is characterized by a neutral reaction of the medium, a high degree of saturation with bases and a humidity of 50-85%. It is stabilized, structural, has a friable consistency, has a high content of humified organic matter (57-75% on a dry basis), nitrogen (N total - 2.5-4% on a dry basis) and phosphorus (P ₂ O ₅ - 2.5-5% on dry matter), with an average potassium content (0.2-0.5%) and low water retention capacity.

The mechanically dehydrated sludge in its composition meets sanitary standards (SanPiN 2.1.7.573-96), and it can be used in agriculture, as is done in many industrialized countries. Worldwide, waste management and environmental protection are a pressing issue. From filter presses, dehydrated sewage sludge is transported outside the cities and stored at landfills. Accumulating in huge quantities, they create a big environmental and economic problem for megacities. In Moscow alone, this amount is more than half a million tons annually. In our country, in agriculture, no more than 4-6% of OGSV is used.

An obstacle to the widespread use of sludge as an organo-mineral fertilizer may be a high content of heavy metals in the form of carbonates, sulfides, organic complexes associated with bacterial biomass (Chebotarev N.T. Problems of wastewater sludge treatment in Moscow // Chemicals in Agriculture . - 1994. - No. 4. - S.21-22). However, according to the conclusion of the All-Russian Research Institute of Fertilizers and Soil Science (VIUA), OGSV Moscow can be used as organic fertilizer without the risk of contaminating the soil and crop products with heavy metals (cadmium, manganese, copper, arsenic, nickel, mercury, lead, chromium, zinc) (table .1) (Sedykh E.M., Adzhienko V.E., Starshinova N.P., Bannykh L.N., Tatsiy Yu.G., Gulko N.I. Analysis of urban sewage sediment // RIA Standards and quality. Partners and competitors. - 2001. - No. 1. - S.16-20).

Table 1 The content of trace elements and heavy metals in the sewage sludge of the Kuryanovskaya aeration station Items The content in OGSV, mg / kg, dry matter MPC Manganese 128-600 2000 Arsenic 3.7-4.2 twenty Mercury 0.17 fifteen Lead 72-84 1000 Copper 344-439 1500 Nickel 48-64 400 Zinc 244-800 40,000 Chromium 304-682 1200 Cadmium 3-5 thirty Molybdenum 2-8 not standardized Cobalt 4-20 not standardized Selenium 2-8 not standardized

Modern urban wastewater treatment technologies also eliminate the epidemiological danger. At present, OGSV is recommended for use in cereals for fodder, fodder and industrial crops, in forestry and nursery farms, in meadow cultivation, seed growing, floriculture, where the probability of heavy metals and pathogens getting into food chains is minimal (Vorobyeva R.P., Dodolina V.T., Merzlaya G.E. Ecologically Safe Waste Management Methods - Barnaul: Publishing House of the Ministry of Agriculture of the Russian Federation, 2000. - 555 p .; EM Romanov. Environmental Aspects of Sewage Sludge Utilization in Forest Nurseries // Protection Problems surrounding th environment from industrial, household, biological and medical waste, sewage sludge: mater international, scientific and practical conference - Penza, 1997. - S.147-150)....

Thanks to the activity of thermophilic bacteria, which are active at high temperatures, fresh sediments are almost sterile, which minimizes the effect of pathogenic microflora on microplants. At a temperature of 55-65 ° C, thermophilic microorganisms predominate and the process of biophysical disinfection of compost proceeds. When the temperature in the compost rises to 65-80 ° С and higher, its thermal disinfection begins (Strauch D. Gesetzliche hygienische Anfordeungen an Komposte aus Abfallstoffen / VDLUFA-Schriftenreihe - Verbandt Dt. Landwirtschaftlicher Untersuchungs - und Forschtstanstad, 1988; - S.983-1002).

As our studies have shown, the positive effect of OHSW on the growth and development of garden plants is also associated with the presence of hormonal activity of this substrate due to the presence of the vital products of microorganisms that accumulate in the sediment during microbiological wastewater treatment (Table 2).

table 2 Hormone content in dehydrated sewage sludge, RGAU-MSHA, 2006 OGSV storage period IAA ng / g mass CC ng / g mass GA ng / g mass ABA ng / g mass (Control) Tn: P = 1: 1 0.05 - - - Fresh OGSV 1.86 60.7 57.4 14.2 OGSV after storage for 1 year 0.65 40,4 67.1 1.9 OGSV after storage for 7 years 0.62 13.5 - - Tn - lowland peat; P - perlite

Fresh sediments after filter presses are characterized by the highest content of auxins (IAA), cytokinins (CC) and gibberellins (HA), hormones that stimulate growth by division and stretching, and control morphogenesis as a whole. When storing sediments at the landfills for more than one year, the content of CK decreases by 1.5, IAA - by 3 times, however, the high ratio of the sum of CK + IAA to ABA and the content of gibberellins positively affect the formation of the accessory root system and assimilation surface. After prolonged exposure to silt sites (up to 7 years), the hormone content in the OGSW samples is significantly reduced, the substrate is again populated by pathogenic microflora, and its structure is deteriorating.

The high nutritional value of substrates containing fresh OGSV, the presence in the environment of hormones synthesized by microorganisms at the stage of thermophilic digestion, the low level of pathogenic microflora provide high value of mixtures for adaptation and growing regenerants.

Another component of the experimental substrate is neutralized sphagnum peat (peat of the upper type of the moss group, pH 5.5-6.5;

producer - peat enterprise “Pelgorskoye-M”) is ideally suited for low-volume technologies. It is characterized by a high content of sphagnum mosses (more than 80%), organic substances (96-97.7%), the absence of heavy metals, good physical properties: uniformity, friability, flowability, low bulk density (130-180 kg / m ³ ), a long fibrous homogeneous structure with a minimum fraction of dust fraction, high gas and water absorption capacity, thermal insulation properties. Sphagnum peat has significant buffering and high sorption and ion exchange ability.

High porosity (90-95%) and moisture capacity favorably distinguish peat from lowland and transitional. Even with abundant watering, peat in the pores contains up to 20% of air, therefore, waterlogging does not occur. This is an important condition for the successful adaptation of regenerants, as excess moisture can cause root rot, stem stems, and root dying.

Optimal physical properties, favorable water-air regime, high water-holding and absorption ability create unique conditions in the root habitat for successful survival, growth and development of in vitro plants. In addition, sphagnum peat produces carbon dioxide, which is necessary for the normal development of regenerants in a limited enclosed volume and in greenhouses (Kuznetsova L.M., Galaktionov A.A. Peat soils. L., Transactions of VNIITP, issue 55, S.- 99-108 .; Kuznetsova L.M., Bulganin V.N., Simonova G.P., Scherbakov V.A. Rationale for the parameters of peat raw materials used in crop production.Processing and use of peat. L., Collection of scientific works of VNIITP 59, S.5-12).

Peat peat also contains natural humic acids, which have a positive effect on the plant and mycorrhiza, as well as amino acids necessary for the conversion of nutrients to an accessible form.

Adding peat to the substrate not only improves its physical properties, but also inhibits the activity of pathogenic microflora Streptomyces spp., Penicillium spp., Mortierella spp., Trichoderma viride spp., Fusarium spp., Rhizoctonia spp., Pythium spp., Alternaria spp. and Botrytis spp. (R. Tahvonen The diseas suppressiveness of light-colored sphagnum peat and biocontral of plant diseases with Streptomyces sp / Acta Horticulturae 342: International Symposium on Horticultural Substrates other than Soil in situ). Peat peat has aseptic bactericidal properties due to the high acidity of the medium and the presence of phenolic compounds (L.M. Kuznetsova. Use of peat in sheltered soil. Peat in agriculture of the Non-chernozem zone: reference book / V.N. Efimov, I.N. Donskikh, L.M. Kuznetsova et al. Compiled by V.N. Efimov. - L .: Agropromizdat. Leningra. Otdniye, 1987. - P.109-130), does not contain technical pollution, pests and seeds of weeds.

The combination of these characteristics makes it possible to create favorable conditions for adaptation and growing ex vitro microplants.

The content of OGSV in the substrate is optimal to achieve the task. With a lower or higher content of OHSW in the substrate, the effect of its use is less significant.

Case Studies

Studies on the adaptation of lilacs and berry bushes were carried out in the fruit growing laboratory of the Russian State Autonomous Agricultural University named after K.A. Timiryazev in 2005-2007. The results presented in tables 3-11 indicate a significant positive effect of adaptation conditions, including artificial substrates, for the timing of transplantation, survival of regenerants, biometric indicators and the yield of high-quality planting material with a closed root system in a short time.

Example No. 1

In the beginning - mid-February, lilac microplants are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban wastewater sludge (OHSW), not more than one year of storage and neutral peat (pH 6-6.5). The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° C and additional exposure from early January to mid-March (illumination 2000 lux, 16 hours), followed by transplantation of plants in containers (V 1.6 l) in early May. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the following ratio:

in control - lowland peat and perlite in equal proportions;

in experimental versions - OGSV 100%; high peat 50-80% and OGSV 20-50%; perlite 50-80% and OGSV 20-50%; high peat 25-40%, perlite 25-40% and OGSV 20-50%.

Table 3 The development of lilac microplants (Lebedushka cultivar) during their adaptation in non-sterile conditions on substrates containing OGSV, 2005 Type of substrate Ratio of components The share of OGSV in the substrate,% The average length of shoots, cm The total area of leaves, cm ² The total length of the roots, cm Control Tn P 1: 1 - 6.2 15.4 74,2 ABOUT - one hundred 8.7 16.3 308.1 About P 1: 1 fifty 10,2 34.1 296.6 About P 1: 3 thirty 9.9 76,4 305.5 About P 1: 4 twenty 7.4 30,2 215.1 About tv 1: 1 fifty 9.2 104.4 347.5 About tv 1: 3 thirty 12,2 145.1 416.8 About tv 1: 4 twenty 14,4 168.9 496.9 About tvp 2: 1: 1 fifty 8.1 52.6 112.1 About tvp 1: 1: 1 thirty 7.5 77,2 254.3 About tvp 1: 2: 2 twenty 10.3 63.8 142.1 O - OGSV; Tn - lowland peat; TV - horse peat; P - perlite

The best for adaptation and growing lilac microplants under non-sterile conditions turned out to be substrates containing horse peat and OGSV (20-30%). On such substrates, the highest yield of viable regenerants and the best development of the aerial part and root system were noted: the length of shoots increased 2.3 times, the total leaf area increased 11 times, and the total root length 6.7 times.

Subsequently, the optimal OGSV content in the substrate was clarified for adaptation and in vitro growth of berry crop plants (Tables 4, 5, 6).

Example No. 2

Blackberry microplants (Thornfrey variety) in early February are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban sewage sludge (WGW) for no more than one year of storage and neutral peat (pH 6-6.5). The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° C and after-exposure (illumination 2000 lux, 16 hours) from early February to mid-March with subsequent plant transplantation in containers (V 1.6 l) in early May. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the ratio: OGSV: peat = 1:20, 1: 9, 1: 4, 1: 3, 2: 3 with a proportion of precipitation in the substrate, respectively, 5, 10, 20, 30, 40 %; (in control - lowland peat and perlite in equal proportions).

High (95-100%) survival rate of test plants under non-sterile conditions on substrates containing 10-30% OGSV (OGSV: TV 1: 9, 1: 4, 1: 3) was noted. Microplants are highly viable and even. Significantly (1.7-2.5 times) the length of the shoots, their leafiness and the total length of the roots increase. The best results were obtained with a ratio of OGSV and high peat of 1: 4.

The effect of the use of substrates containing OGSV and horse peat in the same ratio is confirmed on yellow raspberries (Yellow giant) and on the raspberry-blackberry hybrid of Tiberias (Tables 5, 6).

Example No. 3

In early February, raspberry microplants (cultivar Yellow Giant) are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban sewage sludge (WGW) for no more than one year of storage and neutral peat (pH 6-6.5) . The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° C and additional exposure from early February to mid-March (illumination 2000 lux, 16 hours), followed by transplantation of plants in containers (V 1.6 l) in early May. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the ratio: OGSV: high peat = 1: 20, 1: 9, 1: 4, 1: 3, 2: 3 with a proportion of precipitation in the substrate, respectively, 5, 10, 20, 30, 40%; (in control - lowland peat and perlite in equal proportions).

Example No. 4

In early February, microplants of raspberry-blackberry hybrid (Tiberberry variety) are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban sewage sludge (WGW) for no more than one year of storage and neutralized peat (pH 6-6, 5). The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° C and after-exposure (illumination of 2000 lux, 16 hours) from early February to mid-March and with plant transplants in containers (V 1.6 l) in early May. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the ratio: OGSV: high peat = 1: 20, 1: 9, 1: 4, 1: 3, 2: 3 with a proportion of precipitation in the substrate, respectively, 5.10.20, 30, 40%; (in control - lowland peat and perlite in equal proportions).

Table 6
Survival, growth and development of microplants of raspberry-blackberry hybrid (Tiberberry variety) upon adaptation in substrates containing horse peat and OGSV (planting on adaptation 1.02; measurements 1-10. 06. 2006) Ratio of components The share of OGSV in the substrate,% Survival rate,% The proportion of well-developed in vitro plants,% Plant height, cm The number of leaves Number of shoots The total area of the leaf surface of one plant, cm ² The diameter of the root neck, mm The total length of the roots, cm TP 1: 1 the control 60.7 one hundred 23,2 13.5 1,0 424.5 3,1 100.0 Answer 1:20 5 95.9 one hundred 40.6 20.1 1,5 725.6 3.2 131.1 1: 9 10 one hundred one hundred 108,2 29.9 2.2 1392.6 3.6 226.2 1: 4 twenty one hundred one hundred 140,4 30.6 2.1 2835.8 3.9 311.9 1: 3 thirty one hundred one hundred 120.3 28.1 2.1 1700,2 3.8 267.4 2: 3 40 one hundred one hundred 60.7 23.5 2.0 752.6 3.0 135.0

The ratio of OGSV and high peat is optimal. With a lower and higher proportion of OHSW in the artificial substrate, biometric indicators characterizing the quality of regenerants, their growth and development are lower.

The next task was to determine the optimal timing of in vitro transplantation of plants on experimental substrates.

Example No. 5

In the first ten days of January, blackberry microplants are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban sewage sludge (WGW) for no more than one year of storage and neutral peat (pH 6-6.5). The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° С and additional exposure from the beginning of January to the end of February (illumination 2000 lux, 16 hours) with subsequent transplantation of regenerants in containers (V 1.6 l) in mid-March. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the control: OGSV: perlite and lowland peat in equal proportions; in experimental versions: OGSV and peat in the ratio of 1: 4.

Example No. 6

In the first ten days of January, blackberry microplants are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban sewage sludge (WGW) for no more than one year of storage and neutral peat (pH 6-6.5). The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° С and additional exposure from the beginning of January to the end of February (illumination 2000 lux, 16 hours) with subsequent transplantation of regenerants in containers (V 1.6 l) in mid-March. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the control: OGSV: perlite and lowland peat in equal proportions; in experimental versions: OGSV and peat in the ratio of 1: 4.

Example No. 7

In the first ten days of January, blackberry microplants are transplanted into seedling cassettes with cells (d 40 mm) filled with a substrate containing urban sewage sludge (WGW) for no more than one year of storage and neutral peat (pH 6-6.5). The first two weeks of microplants in cassettes are kept under airtight film shelters on racks with expanded clay (layer 2-3 cm) at a temperature of 22-26 ° С and additional exposure from the beginning of January to the end of February (illumination 2000 lux, 16 hours) with subsequent transplantation of regenerants in containers (V 1.6 l) in mid-March. The control substrate before planting microplants is treated with 1% euparen solution; experimental substrates do not pre-disinfect and do not steam; all substrates are prepared by simple mixing of the components in the ratio: OGSV: in the control, perlite lowland peat in equal proportions; in experimental versions: OGSV and peat in the ratio of 1: 4.

Table 9 Survival, growth and development of microplants of raspberry-blackberry hybrid (Tiberberry variety) upon adaptation in substrates containing horse peat and OGSV depending on the timing of transplantation into non-sterile conditions (measurements 1-10.06.2007) Landing for adaptation Survival rate,% The proportion of well-developed in vitro plants,% Plant height, cm The number of leaves Number of shoots The total area of the leaf surface of one plant, cm ² The diameter of the root neck, mm The total length of the roots, cm control of consumer goods 1: 1 1st of January 30.6 29.8 25.1 5.1 1,0 35.8 2,8 75,4 January 15 34.8 30.1 30.8 7.4 1,0 350.6 2,8 88.5 1st of February 59.9 35,2 28,4 7.0 1,0 349.5 2.9 92.3 February, 15 66,2 40.1 30,2 8.5 1,5 380.5 2,8 93.5 28th of February 87.6 55,4 38.6 8.1 1,0 400.6 3,1 96.2 March 1 85,4 55,2 45,2 11,2 1,0 415.4 3.0 106,4 March 15th 90.2 65,4 55.1 13.0 1,5 410.2 3.4 121.5 experimental substrate - O: TV 1: 4 1st of January 75,2 87.4 100.3 23,2 1,5 1350.0 3,5 215.4 January 15 73.7 88.3 104.5 25.6 1,5 1355.4 3,5 216.8 1st of February 80.5 99,4 108,5 26.1 1,5 1352.6 3,5 219.5 February, 15 one hundred 98.9 115.4 25.3 2.0 1368.2 4.2 230.8 28th of February one hundred one hundred 120.3 30.8 2.0 1370.6 4.3 250.8 March 1 one hundred one hundred 119.4 28,4 2.2 1385.4 4.1 248.5 March 15th one hundred one hundred 108,5 25.1 2.2 1356.2 4.0 230,4

The use of OGSV and neutralized peat in the composition of the substrates in a 1: 4 ratio allows plants to be transplanted in vitro to non-sterile conditions at an earlier date (early January) than by conventional technology (early to mid-March). Due to the presence of gibberellins in the environment, regenerants on a short day actively grow and develop. The race (2.5 months) in growth and development makes it possible in mid-March to transplant meristem plants into containers, and by the beginning of the growing season (early May) receive high-quality planting material with a closed root system.

Claims (2)

1. The method of growing in vitro plants of berry and ornamental shrubs in non-sterile conditions, including the transplantation of microplants from test tubes into cassettes filled with an artificial substrate, characterized in that dehydrated stabilized sludge from urban wastewater and neutralized peat in a ratio of 1: 4 are used as a substrate , while the transplant is carried out in early January. 2. The method according to claim 1, characterized in that the microplants transplanted into the cassettes are grown under a sealed film shelter for two weeks.