SU1738170A1 - Method for microclonal propagation of virus-free potatoes - Google Patents

Abstract

Use: agriculture, potato, biotechnology, seed production. SUMMARY OF THE INVENTION: A virus-free potato is propagated by growing a soil mixture at 10-15 klx on a sterilized calcination and providing a water regime by irrigating or flooding the substrate with distilled water of plants obtained from meristem, after which they are grafted and rooted.

Description

The invention relates to the field of biotechnology, in particular to the field of microclonal propagation of plants.

In the practice of microclonal propagation, sterile artificial nutrient media are used to cultivate plants, which makes the technology laborious, expensive and material-intensive. Plant cultivation is carried out in greenhouses with artificial light.

The closest way to replication of virus-free potatoes is the technology that involves rooting micrografts in a dough culture on an artificial nutrient medium consisting of mineral nutrients, plastic substances and growth regulators.

However, chemically pure, expensive reagents, special equipment are used to prepare the media. The cultivation of the aryclones is carried out in the mode of rigid sterility, for which

media autoclaving is used. This makes the technology labor-consuming, material-intensive and expensive.

The purpose of the invention is to simplify the technology of microcloning of virus-free potatoes, reducing its material and labor intensity.

The method is carried out as follows.

The artificial nutrient medium is replaced by thermally disinfected soil mix used for potted crops. For complete destruction of bacterial and viral particles, the soil is kept for 2 hours in a thermostat at 200 ° C. Providing plants with water is achieved by moistening the substrate until full moisture is obtained through transplantation of cuttings into test tubes, as well as additional irrigation on days 6–8 of cultivation. Additional watering can be avoided by placing the tubes at the bottom, under the substrate, a cotton swab and,

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filling this volume with water, which will create enough reserve for the cultivation of the meriklon. Meriklon water supply is provided by distilled water irrigation. The absence of plastic substances in the medium makes it necessary to enhance the photosynthetic activity of the arylones.

In this connection, the illumination in the cultivation building increases to 10-15 klx. The illumination varies, as the cells are differently removed from the light source. After 18–23 days, the meriklon forms 5 internodes and fills the entire volume of the tube. Preparation of the cuttings is carried out without removing the uterine plant from the tube, cutting them with a scalpel for eye microsurgery. The root stump is left in the tube for the second generation of cuttings. 3-4 generations of cuttings can be obtained from one mother plant.

Thus, in comparison with the known method, the proposed method eliminates the use of reagents, including those that are scarce, used for the preparation of artificial nutrient media, in particular: agar, sucrose, a number of mineral salts, vitamins, hormonal growth regulators.

In this connection, there is no need for equipment for dissolving, introducing reagents and sterilizing media: glassware, magnetic stirrers, heaters, metering devices. pH meter autoclave, etc.

Preparation of soil mixture requires much less labor costs than the preparation of artificial nutrient media.

All this taken together reduces the unit cost of production from 0.8–1.3 rubles to 20–25 kopecks.

Example 1. A soil mixture consisting of wood soil, sand and peat in a 2: 1: 1 ratio is prepared for the cultivation of the marilones.

Test tubes, cotton plugs, tampons and the soil mixture are sterilized in a thermostat for 2 hours at 200 ° C. In a sterile box, a cotton swab, 1.5-1.8 cm3 in volume, is placed at the bottom of the tube, this volume is filled with a fresh distillator, and another 2.2-2.5 ml of distilled water is added. Then pour 3.5 - 4 g of sterile substrate. (2.8 -3 cm. Close with a cotton plug and infuse for 30 - 40 minutes.

Adretta maternal plants are cut into cuttings in the middle of the interstice. The lower node is left on the mother plant, topped up with 1.7-2.3 ml

The distillate and mother plants are left to pivot the generation of cuttings.

Cuttings with long tweezers are placed in the substrate so that the node is at a depth of 3-5 mm, the tube is closed with a stopper.

Test tubes with cuttings are installed on light racks with a combination of lighting: a vertical row of luminescent lamps and an M1X-H lamp located on top. Illumination 15 klk, lighting around the clock. The air temperature during the growing season is 18–20 ° C, the humidity is 50–60%. The rooting of the cuttings was 84%.

Example 2. A soil mixture is prepared from humus soil, sand and peat in a ratio of 2: 1: 1. Sterilization was carried out as described above. Test tubes are filled with 3.4-4 g of dry substrate, 2.2-2.5 ml of distilled water is poured, closed with a stopper and drawn for 30-40 minutes.

Used meriklon varieties Nevsky. The technology of grafting and growing plants is similar to that described above, the illumination of 10 klx. The rooting rate of cuttings was 90%.

Example 3. A sterile soil mixture was also used in wide-bottomed dishes, in particular, in three-liter bottles. The substrate is poured into sterile dishes - domestic black earth with sand (2: 1) up to 5–7 cm high, watered at the rate of 0.65– 0.75 ml of distillate per 1 g of substrate. The cuttings of the Izor variety are planted according to the 2.5 x 2.5 scheme with concentric circles. The opening of the flask is closed with a cotton-gauze filter. Cultivated under the same conditions, with the exception of an illumination of 12 klx, the rooting of the cuttings is increased to 95%, which is caused by an improvement in the conditions of substrate aeration.

The enzyme immunoassay shows the persistence of the quality of the arylones in the described methods of replication.

The time for the formation of five interstices by the marilones is 18-20 days, which corresponds to the prototype.

The use of the proposed method of replication of virus-free potatoes provides the following advantages as compared with the known methods: the material consumption of the technology is significantly reduced; reduced labor costs; reduced production costs; simplified technology.

Claims (1)

The invention The method of microclonal propagation of virus-free potatoes, including growing on a sterile substrate, full of plant meristem, their cherenko f the rooting of the obtained cuttings, 200 ° C for 2 hours, the soil mixture, which is different from the fact that, with the aiming, the micro-cloning technology of 10–15 klc is illuminated when the neither, reducing its material and labor-required water regime by irrigation or bone, as a substrate, is used5 by flooding the substrate with distilled sterilized by calcination drive.