RU2762417C1 - Method for clonal micropropagation of sarmatian bellevalia (bellevalia sarmatica (pall.ex georgi) woronow) - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. The invention is a method for propagation of plants of Belvalia Sarmatian B. sarmatica by in vitro culture, including sterilization of seeds, planting on nutrient media, reproduction of microbulbs, their subsequent rooting, adaptation of regenerated plants to ex vitro conditions. After adaptation to non-sterile conditions and growing, standard Sarmatian Belvalia seedlings with a closed root system are obtained.

EFFECT: invention makes it possible to obtain a large amount of leveled environmentally friendly planting material.

1 cl, 5 dwg

Description

The invention relates to the field of plant biotechnology and can be used for the mass production of high-quality planting material of Sarmatian Belvalia (Bellevalia sarmatica (Pall, ex Georgi) Woronow), a plant listed in the Red Book of the Russian Federation and endangered (rare status - 2 - decreasing in number) [2]. The resulting plants can be used to replenish the introductory collections of botanical gardens and return to natural habitats, as well as for landscape gardening. EFFECT: invention makes it possible to obtain a large amount of leveled environmentally friendly planting material.

The proposed invention is new, since no solution has been found in the patent and scientific and technical literature with the claimed set of distinctive features.

Belvalia Sarmatian is a perennial bulbous plant. Height up to 20 to 30 cm. Leaves are belt-shaped, concentrated at the base of the stem. It blooms with a multi-flowered brush, the color is gray-brown, the pedicels are drooping, elongate when the fruit ripens. The fruit is an oblong capsule with 2-4 seeds. Blooms in spring. Propagated by seeds, blooms in the 5th year after sowing. Belvalia Sarmatian in nature reproduces only by seeds, data on vegetative reproduction are absent [2, 3].

All this makes it necessary to conduct research on the development of a technique for clonal reproduction and determines the relevance of this work [1, 5, 6 |.

The use of clonal micropropagation makes it possible to obtain a homogeneous planting material, to shorten the cultivation period of the culture, and also provides an opportunity to carry out work throughout the year and save the area required for growing Bellevalia sarmatica planting material [4, 5].

The invention is a method for propagating Bellevalia sarmatica (Pall, ex Georgi) Woronow plants by in vitro culture. including sterilization of seeds, planting on nutrient media, reproduction itself (Кр = 7.8), subsequent rooting and adaptation of microbulbs to ex vitro conditions. After adaptation to non-sterile conditions and growing, standard Sarmatian Belvalia seedlings with a closed root system are obtained. The result of the invention is a leveled and improved planting material - 15,000 pieces.

Progress

In the course of research, a method of clonal micropropagation of Sarmatian Belvalia was developed.

The method is implemented as follows.

Seeds were used as primary explants.

Optimal sterilization system: 1.) fungicide - 30 minutes: 2.) 70% ethanol - 2 minutes; 3.) 7% calcium hypochlorite - 20 minutes.

At the stage of micropropagation proper, the effect of growth regulators and their concentrations on the reproduction rate was investigated. The highest values of morphometric parameters were obtained on nutrient media containing 10 mg / L 6-BAP with the addition of 0.1 mg / L NAA. A further increase in the concentration of growth regulators did not give a positive effect (Fig. 1). The maximum multiplication factor of B. sarmatica, equal to 7.8 ± 1.95, is achieved when cultivated on a nutrient medium containing 6-BAP (10 mg / L) and NAA (0.1 mg / L).

Under laboratory conditions, microshoots of B. sarmatica were grown under artificial illumination (3000 lx) and a photoperiod of 16/8 hours at a temperature of 23–25 ° C and a humidity of 70%.

MS, 1.5 мг/л ИМК и 20 г/л сахарозы, при температуре 16-18°С и освещенности 500 лк (фиг. 2).The maximum percentage of rooting (76.5 ± 1.2) was obtained on a nutrient medium containing

MS, 1.5 mg / L BCI and 20 g / L sucrose, at a temperature of 16-18 ° C and illumination of 500 lux (Fig. 2).

During adaptation, regenerant plants were planted in a soil substrate previously sterilized at 85–90 ° C for 1–2 h. The substrate consisted of a mixture of high-moor peat, sand, and perlite (1: 1: 1).

The different stages of clonal micropropagation of Sarmatian Belvalia are shown in Fig. 3-5: Figs. 3 - the stage of initiation, FIG. 4 - the stage of actual micropropagation, FIG. 5 - the stage of rooting.

Sources of information

1. Molkanova OI .. Shirnina. I.V. and Mitrofanova. I.V. Conservation and micropropagation of rare and endemic species in genepool collections of the Russian Federation. J. of Biotechnology. - 2018. - P. 83-84.

2. Red Data Book of the Russian Federation (plants and mushrooms). - M .: KMK Scientific Publishing Association. - 2008 .-- 855 p.

3. Kurdyukova ON Seed productivity and germination of seeds of early flowering plants // Bulletin of the State Nikitsky Botanical Garden. 2020: (136): 55-60.

4. Kutovsky-Mushtuk A.I., Chorkina N.G., Lozinskii M.G. Conservation of the rare species Bellevalia sarmatica (Pall, ex Georgi) Woronow culture in vitro // Coll. mat. the annual meeting of the Society of Plant Physiologists of Russia, the All-Russian Scientific Conference with international participation and the school of young scientists. In 2 parts. Irkutsk: 2018 .-- pp. 1295-1298

5. Magulaev A.Yu. On the reproduction biology of Bellevalia sarmatica (Georgi) Woronow under conditions of introduction // Bulletin of the Voronezh State University. Ser .: Geography. Geoecology. - 2011. - Ν 1. - S. 165-166: tab. - Bibliography: p. 166

6. Molkanova OI .. Gorbunov Yu.N., Shirnina IV .. Egorova DA Application of biotechnological methods to preserve the gene pool of rare plant species // Botanical Journal. - 2020. - T. 105. - No. 6. - S. 610-619.

Claims (1)

The method of clonal micropropagation of Bellevalia sarmatica (Pall, ex Georgi) Woronow) includes sterilization of seeds with 7% calcium hypochlorite (exposure for 20 minutes), propagation itself on MS nutrient media containing 10 mg / l 6-BAP and 0.1 mg / l NAA, their subsequent rooting on the medium

MS, 1.5 mg / l BCI and 20 g / l sucrose at a temperature of 16-18 ° C and illumination of 500 lux, adaptation of regenerated plants to ex vitro conditions.

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