RU1792269C - Method for plant adaptation for non-sterile conditions - Google Patents

Abstract

Usage: biotechnology, in particular with microclonal propagation of 2 plants. The inventive method includes the selection of regenerated plants obtained in tissue culture, and plants are selected that have 3-5 leaves and at least 2-3 roots 1.5-3 cm long. After that, the regenerated plants are treated sequentially with 1% potassium permanganate solution for 3-5 min and a solution of indolylacetic acid at a concentration of 10-15 mg / l, then the plants are planted on a substrate in peat pots and placed under controlled conditions under illumination of 8-10 thousand microliters and a 16-hour photoperiod cover with transparent mater Ial, which is removed after 7-10 days, 6 tablets. THX

Description

The invention relates to agriculture, in particular to plant propagation methods. The invention can be used in viticulture for accelerated propagation and improvement of promising grape varieties, in studies on the physiology of grapes. Serves the environment,

A known method of adapting plants propagated in tissue culture to non-sterile conditions in which adaptation takes place in two stages. First, rooted tube plants are transplanted onto a liquid nutrient medium. Then, after a week, the plants are transplanted into an autoclaved soil mixture and covered with glass vessels. After peeling, the plants are planted in a greenhouse to continue adaptation.

The disadvantage of this method is the significant labor costs, as well as the use of expensive nutrient medium. .

There is a method of adaptation of plants rv propagated in tissue culture to conditions of UV open ground, when for this purpose film greenhouses are used, which create specially controlled conditions with high relative humidity of air and shading from direct sunlight over a large area of greenhouses.

The disadvantage of this method is that it is expensive and complicated.

A known method of adaptation of plants propagated in tissue culture, to nest. conditions, including selection of regenerated plants, preparation of a substrate, planting of regenerated plants in a substrate,

coating them with transparent plastic material and keeping the environment under controlled conditions, followed by removal of the coating;

The disadvantages of this method are the insufficiently high quality and yield of seedlings.

The aim of the invention is to improve the quality and increase the yield of seedlings. . J -;

This is achieved by the fact that in the method of adaptation of plants propagated in tissue culture to non-sterile conditions, including selection of regenerated plants, preparation of the substrate, planting of the regenerated plants in the substrate, coating them with transparent plastic material and keeping under controlled conditions an external media with subsequent removal of the coating according to the invention, regenerant plants are selected having 3-5 leaves, at least 2-3 roots 1.5-3 cm long, which are sequentially treated with a 1% solution of potassium permanganate, and then for 3-5 minutes, indolylacetic acid in a concentration of 10-15 mg / l, after which they are planted in a substrate in a peat-pot, then the plants are kept under controlled conditions under illumination of 8-10 thousand lux and a 16-hour photoperiod, and remove the coating after 7-10 days.

A comparative analysis of the claimed solution with the prototype showed that a new method in the claimed method is that the adaptation is carried out in one step. Plants from test tubes are planted in order to adapt them to non-sterile conditions in peat pots with a substrate - peat: soil: sand, plant adaptation is carried out in individual chambers, and plants are transplanted from test tubes in the presence of 3-5, not 4 -10 leaves, which reduces the preparation of plants for adaptation by 20-30 days. The novelty also consists in treating plants after removing them from test tubes and before planting in peat pots 1% solution of potassium permanganate and a solution of indolylacetic acid (IAA) in a concentration of 10-15 mg / l for 3-5 minutes. Adaptation of plants is first carried out in closed individual chambers of transparent plastic material in peat pots for 7-10 days, and then 15-20 days in peat pots without coating. After the plants have fully adapted, they are transplanted into the greenhouse. When conducting a search in the patent and scientific literature, a method for growing plants in peat pots was discovered. However, in the known method, plants are planted in a peat pot not for the purpose of preliminary adaptation (intermediate link) before final planting in the soil, but for direct growth in a peat pot, as in the soil, which is a significant difference.

The proposed method is characterized in that it is aimed at increasing the resistance of plants to stress factors when transferring them from test tubes to non-sterile conditions. The first stressor is the possibility of infection. To avoid this, plants are treated with a 1% potassium permanganate solution. Bags are also treated with this solution. A pre-sieved and moistened substrate avoids damage to the delicate root system. And treatment with a solution of indolylacetic acid promotes the laying of new root primordia. When a plant is planted in a peat pot, roots and plants are attached to the substrate, ensuring plant resistance, which remains until a standard seedling is obtained, which facilitates transportation to the planting site and completely eliminates damage to the root system. Also, plant growth arrest after transplantation is prevented. In addition, with the possible germination of the roots from the substrate medium, they also enter the nutrient medium of the peat pot, where they will additionally receive the substances necessary for growth and development. In the known method, the growth of the root system of plants is limited to a plastic bag and therefore its inhibition is possible. A stable root system of seedlings helps to improve the quality of plants; accordingly, the yield of finished products also increases, i.e. the goal is achieved. The use of peat pots after removing them from the plastic chambers due to their porosity provides oxygen access to adaptive roots, which is necessary for root formation and root growth, since this consumes a significant amount of energy released during their respiration.

. An improvement in the air-gas regime is also significant because the first ten days of the plant are in closed individual chambers and then in open peat pots. Thanks to the air traffic

masses in the room, plants have the ability to extract the carbon dioxide they need from the atmosphere, which leads to increased photosynthesis and improved plant development. The created illumination in 8-10 thousand lux and a 16-hour photoperiod contributes to the improvement of root formation, photosynthesis, and regulation of stomatal conduction. Intensive exposure to light is a trigger for the production of endogenous auxins, which provide the optimal ratio and development of the aerial part and the root system of the plant. The plants adapt better in high light conditions, while the photosynthetic apparatus becomes highly active and is protected from premature aging due to the active synthesis of cytokinins in the light, which affects the biological and economic yield of plants .;

The essence of the invention will be clear from the following description of an example of a specific implementation of the method.

The method is carried out as follows. ;:. .

Clone-propagated test tube grape plants are selected. Plants should have 3-5 leaves, at least 2-3 roots with a length of at least 1.5-3 cm. Plants with a large number of leaves tolerate adaptation to non-sterile conditions worse, plant survival is reduced.

Then prepare and sterilize the substrate. Peat, sand, soil are sieved through a sieve of 5 mm diameter, taken in equal parts by volume (1: 1: 1), mixed thoroughly and placed in enameled trays. Prepared substrates and empty peat pots are installed in the boiler of the GK-100-3 sterilizer and sterilized for 15-20 minutes at a pressure of 0..1-0.15 MPa and a temperature of 115-120 ° C. At the end of sterilization and after the substrate has cooled, it is placed in peaty pots and poured with a 1% solution of potassium permanganate for 30-40 minutes, so that the soil is well saturated. After this time, the soil is ready for work.

Packages 18-20 cm wide and 40-45 cm long are prepared in advance from a plastic film. Before planting, they are washed in distilled water with the addition of manganese. When everything is prepared, proceed to transplant the plants. Plants are removed from the test tube with tweezers, washed from agar-agar and placed in a bath with a previously prepared 1% solution of potassium permanganate, and then for 3-5 minutes in a bath with a solution of indolylacetic acid (IAA) in a concentration of 10-15 mg / l

5 From the bath, the plants are transferred to a peat pot, where a recess has been made in the substrate with other tweezers, melt them, fall asleep and lightly press down so that the plant assumes a vertical position. Then, a bag made of plastic material (individual chamber) is put on a peat pot with a plant, which is connected from the bottom with a paper clip. Planted plants are placed in a wooden box where they place a label and put them for further growth on shelves of accelerated plant growing (IWRM). In the SU In Re, the plants are at a temperature of 25-27 ° C, illumination of 8-10 thousand lux, and a light period of 16 hours. After 7 days, the packets are removed, the plants are watered with distilled tap water with the addition of (up to 1% solution) manganese

5 potassium. Plants, as before, are exposed to artificial light, ultraviolet radiation of which affects open plants unhindered. Due to exposure to light

0 a comfortable ambient temperature is created, significant growth processes occur.

After a week, the plants are fed a solution of mineral elements

5 V.A. Chesnokov. Adaptation of plants at SUVR is carried out within 22-30 days, and then the plants are planted in a peat pot in a film greenhouse with partially controlled conditions:

0 furrow irrigation, airing by opening doors, shading by whitening the film with lime.

The optimal time for planting plants in the film greenhouse was established - end 5 ma -media Jun, permissible - the first decade ma-the last decade jun. There are no special nutritional mix builds in the greenhouse. Preliminary site surface carefully leveled

0 grader or bulldozer, in the fall they raise the plantation to a depth of 45-50 cm, followed by careful cultivation. In a film greenhouse with a width of 3.5 m, the area prepared for planting is broken down with

5 using twine to five equal rows.

The distance and row between the seedlings is 10 cm. At the same time, 1500-2000 seedlings are planted in the greenhouse. Two people land. With a shovel, dig a landing pit measuring 25x25 cm.

We carefully set the peat pot on the bottom of the landing. sprinkle with soil to the top of the pot and watered. A peg is placed near the plants to which it is tied.

Further plant care, including frequent watering and loosening, the application of mineral fertilizers, the fight against diseases and pests, and shoot gartering, is carried out according to known technology.

Seedlings planted in the two extreme rows (200 pieces) are not dug up, leaving them in a constant place to create a uterine bed. Seedlings planted in three middle rows are dug up, laid for storage, and the next year they are used to plant grape stands.

During the implementation of the proposed method for adaptation to non-sterile conditions, the following physiological processes occur in the plant. . : /.,.,;. ; ,,

When plants propagated in tissue culture (in vitro) are transferred to non-sterile conditions, they experience stress reactions. tion aimed at protecting cellular structures and eliminating adverse changes in cells. That is, a phenomenon of adaptation syndrome is observed.

The process of plant adaptation is intended to do so. to avoid water stress when transferring a plant from a test tube with a nutrient medium (with 100% humidity) to a moist substrate, also to maintain the sterility of the planting material and to improve the further development and growth of strong (upon adaptation) and healthy seedlings. : -:::; ;. . :.; V: H; : ....

. Under unfavorable conditions: in plants, the production of ethylene and abscisic acid (ABA) increases sharply, reducing metabolism, inhibiting growth processes, and contributing to the transition of the plant organism to a resting state. At the same time, the content of auxin, zitokinein and gibberellins in tissues decreases. In the adaptation process, the root formation process plays an important role. In the proposed method, at the first stage, conditions are created for the predominant growth of the root system with slow growth of the aerial part of the plant. This is facilitated by pretreatment with indolylacetic acid, as well as the fact that the plants are covered with a film in a peat pot with a moistened substrate, in which the roots are located, which, like in a test tube, are in a KBM Fort medium on an SUWR at a temperature; contributing to root formation. Thus, the plants are first rooted,

become stable, which is important. At the same time, the amount of carbon dioxide under the film is sharply limited. In view of this, the stomatal fissures of the stem are weakly open and

this part of the plant develops more slowly, more inhibited. At this stage, the nutrients of the substrate are used to a greater extent by the roots. During adaptation, it is unacceptable to remove the plastic bag earlier than 7 days or later 10 days. If the bag is removed earlier, that the plant does not adapt; completely reduce air humidity, because moisture will evaporate intensely. with from the substrate. If removed later, the plant

will be inhibited / from the lack of moisture, which will already be used, and from the absence

carbon dioxide needed for photosynthesis. When the package is removed on time, the stomatal fissures of the stem open

wider, the abundance of carbon dioxide in the air and the permeability of peat pots stimulate photosynthesis, thereby causing active growth now

aerial parts of the plant,; l;

After the package is removed, the open plant is exposed to intense light, which is the starting effect for the production of endogenous auxins, providing optimal

correlation and development of the aerial parts and

root system of a plant. : Y /; -: -

The table shows that the proposed

adaptation method increases the yield of finished

seedlings, chtr helps to reduce their

sebest1I (u1o with d7 kpb honors (; its), namely: the ripening of the shoots, the number of roots improves them; gachestvo. That is, get seedlings with more; high biological

svrst in and, with .. about vtn1m and nezh m with the phytosynthetic device; which, in a critical account, may favorably affect the development of plants grown from nyh; and biological and economic crops

win6grad.;: /: - -. f --1. -.

& o r mula And zobret e and; : A method of adapting plants to non-sterile conditions, including the selection of regenerated plants obtained in a Tissue culture, preparation of a substrate, planting of regenerated plants in a substrate, coating with transparent material and keeping under controlled conditions, followed by

removal of the coating, with the exception of the fact that, in order to improve the quality and increase the yield of seedlings, regenerated plants are selected that have 3-5 leaves and at least 2-3 roots with a length of 1.5- 3 cm, the selected plants are treated sequentially with a 1% solution of potassium permanganate for 3-5 min and a solution of indole-acetic acid at a concentration of 10-15 mg / l, the planting of regenerated plants in the substrate is carried out

in peat pots, when kept under controlled conditions, they provide illumination of 8-10 thousand lux with a sixteen-hour photoperiod, and the coating is removed after 7-10 days.

 ::; -; 1st; / I - T abl and tsa 3

Influence of the primary parameters of plants on further growth and development

: V; -V b y ;; ---:. ;: th; :: l: agzhentsev Agatha; Don: ;;. .-...: ..;.:.;: ;;

Table 1

Table 2

eleven

Influence of the time spent by plants in a closed chamber on plant quality indicators.

Technical and economic advantages of the proposed technical solution to the problem compared to the prototype - a method of adaptation

plants propagated in tissue culture, to non-sterile conditions (Guzun N.I., Litvak A.I. et al. Microcloning of grapes. In collection: Plant cell culture and biotechnology / edited by Butenko R.G. 1983, p. . 117-118) ..

1792269

12 Table 4

Table 5

Table b

Continuation of Table 6