RU2564123C1 - METHOD OF REPRODUCTION OF PEPEROMIA in vitro - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention is a method of reproduction of peperomia in vitro, comprising separating the explants, sterilisation, planting on the Murashige and Skoog culture medium, micropropagation by separating the shoots from the explants, their rooting them on the Murashige and Skoog culture medium and further rooting in the ground, where in micropropagation the explants are planted in an agar Murashige and Skoog culture medium supplemented with 10 g/l glucose and 0.1 mg/l Ribav-Extra and cultured for 4 weeks, then the formed plant-regenerates with roots are soaked in the solution of 3 mg/l indoleacetic acid for 15 minutes rooted under ex vitro conditions in the ground, consisting of a mixture of peat and sand of 1:1.

EFFECT: invention enables to simplify the method of micropropagation by the formation of the root system in the shoots formed in vitro at the stage of micropropagation.

2 dwg, 1 tbl

Description

The invention relates to the field of floriculture and can be used for micropropagation of peperomy plants in vitro.

There is a method of propagating peperomia in vitro (see: Ahmadabadi Μ., Bock R. Development of a highly responsive leaf-based regeneration system for Peperomia species // Turkish Journal of Botany. - 2010. - V. 34. - P. 329- 334). The tops of shoots 1-2 cm long were used as the primary explant, which were sterilized in 70% ethanol for 30 seconds, then washed with sterile distilled water. After that, they were planted on Wednesday Murasige and Skoog. Leaves were separated from the formed young sterile plants, cut into 3 × 3 mm segments and cultivated on Murasige and Skoog medium supplemented with 20 g / l sucrose and growth regulators 3 mg / l benzylaminopurine, 0.15 mg / l indolylacetic acid and 0.014 mg / l gibberellic acid. To form the roots, sterile shoots were transferred to a hormone-free medium with half the concentration of MS salts. Organogenesis was induced in a light room at a 16/8 hour photoperiod with illumination with white fluorescent lamps with an intensity of 2500 lux at a temperature of 25 ° C and 20 ° C, respectively.

The disadvantage of this method is the lack of roots in the resulting shoots directly at the stage of micropropagation, which complicates and lengthens the process of clonal micropropagation due to the need to use the stage of rooting of plants in vitro.

The technical result consists in simplifying the method of micropropagation due to the formation of the root system in shoots formed in vitro already at the micropropagation stage with the positive influence of the optimal glucose concentration and growth regulator.

The technical result is achieved by the fact that the method includes the separation of explants, sterilization, landing on a Murasige-Skoog medium, micropropagation by separating shoots from explants, their rooting on a Murasige-Skoog medium, and further rooting in the soil. During micropropagation, the explants are planted on a Murasige-Skoog agar medium supplemented with 10 g / L glucose and 0.1 mg / L Ribav-Extra, then the formed regenerant plants with roots are soaked in a solution of indolylacetic acid 3 mg / L for 15 minutes and rooted under ex vitro conditions in a soil consisting of a mixture of peat and sand 1: 1.

The method is as follows. The tops of shoots 1-2 cm long are sterilized in 70% ethanol for 30 seconds, then washed with sterile distilled water. After that, they are planted on Wednesday Murasige and Skoog. The shoots are separated from the obtained sterile peperomia plants under aseptic conditions, divided into segments (part of the shoot containing a leaf and axillary bud) and planted in test tubes on an agarized nutrient medium Murashige-Skoog, including 10 g / l glucose and growth regulator - 0.1 mg / l Ribav-Extra. The composition of the medium used in the cultivation of microprobe is standard for micropropagation of plants in an in vitro culture, except for the glucose concentration selected in preliminary experiments (Fig. 1) and growth regulator (Table 1). Cultivation is carried out under continuous illumination with fluorescent lamps with an intensity of about 2500 lux and at a temperature of 25 ° C (physical conditions of cultivation are close to optimal for cultivating plant explants). When microboots are grown on an agarized Murashige-Skoog medium with the addition of 10 g / l glucose and 0.1 mg / l Ribav-Extra within 4 weeks of cultivation, 100% rooted shoots are formed (Table 1). Intense rhizogenesis was observed in all variants of Ribav-Extra media, which is important for further adaptation of plants to ex vitro conditions. Subsequently, the formed plants with roots are soaked in a solution of 3 mg / l of indolylacetic acid for 15 minutes (Fig. 2) and transferred for rooting to ex vitro conditions in a soil consisting of a 1: 1 mixture of peat and sand. After rooting, propagated plants are grown in the usual way.

As a result of using the proposed method, the micropropagation process is reduced by one stage, in addition, 100% rooted shoots are formed regardless of the seasonality of the donor plant taking, which are 100% established in ex vitro conditions.

Thus, in comparison with the known solution, the proposed one allows to simplify peperomia micropropagation due to the formation of the root system in shoots formed in vitro already at the micropropagation stage under the positive influence of the optimal glucose concentration and the growth regulator Ribav-Extra. And also the survival rate of ex vitro regenerated plants is increased due to the pre-planting treatment (soaking) of their root system with a solution of indolylacetic acid (IAA).

Claims (1)

A method of propagating peperomia in vitro, including the separation of explants, sterilization, landing on a Murasige-Skoog medium, micropropagation by separating shoots from explants, rooting them on a Murasige-Skoog medium, and further rooting in the soil, characterized in that the explants are planted on micropropagation Murashige-Skoog agar medium supplemented with 10 g / L glucose and 0.1 mg / L Ribav-Extra, and cultivated for 4 weeks, then the formed regenerated plants with roots are soaked in solution 3 mg / l indolylacetic acid for 15 minutes and root under ex vitro conditions in a soil consisting of a 1: 1 mixture of peat and sand.

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