RU2670485C1 - Method for propagation of immune-resistant potato samples in vitro on aerohydroponic culture - Google Patents

Abstract

FIELD: biotechnology.SUBSTANCE: invention is a method of propagation of immune-resistant potato samples in vitro on aerohydroponic culture, including irrigation of bases of cuttings of regenerants, wherein the aqueous preparation Floron in a concentration of 0.5 % with the addition of pectin in a concentration of 0.3–0.4 % aqueous solution Floron is introduced as a nutrient solution, wherein pectin is prepared from potato parings of healthy tubers of the same variety or a hybrid of regenerants being reproduced.EFFECT: invention provides the simplified technical solution and propagation of healthy material free from viruses.1 cl, 2 tbl

Description

The invention relates to the field of biotechnology and can be used in the reproduction of the immune source material of potatoes for further seed production.

There is a method in which clonal micro-propagation of plants is carried out in an in vitro culture by microgravating regenerants and rooting cuttings on a nutrient medium, in which fructose is used as a carbon source for acceleration (patent No. 20580780 published 10.06.1997, IPC AO1H 4/00) .

However, the cultivation of the cuttings is carried out in heterotrophic conditions, which lengthens the time for obtaining regenerated plants in comparison with autotrophic cultivation conditions. For the cultivation of plants in test tubes, lighting in the range of 20-30 volts is necessary. At the same time, the nutrient medium must contain a sufficient amount of organic compounds, vitamins, hormones and trace elements. Biologically active components can bring to genetic changes in the process of reproduction in vitro.

There is also known a method in which a disease-resistant starting material is obtained, where the cultivation is carried out on nutrient media of a specific composition (patent No. 2524424, published on July 27, 2014, bulletin No. 21 of the IPC AO1 4/00, AON 1/00, AON 5/00 ).

However, the known method is complicated by the fact that the regenerants are cut off and transferred to another medium, where a certain temperature and light are observed, followed by the selection of disease-resistant forms, which complicates the method.

The known method, where regenerants are propagated by introducing into the solution glycine, activated carbon and ascorbic acid (patent No. 2329639 published on July 27, 2009, bulletin No. 21, CIM AO1H 4/00).

In this way, the samples are propagated at different temperatures and illumination, which also complicates the method.

A technical solution is known in which the reproduction of regenerants is carried out by etiolizing seedlings by sterilization in 0.1% diancid solution for 3-5 minutes and further washing with water, in addition, sucrose, ferulic acid, kinetin, thiamine are introduced into the culture medium of tuberization No. 2632938, published 11.10.2017, Bull. No. 29, CIM AO1N 4/00).

This solution is quite complex and costly to create a nutrient medium with the inclusion of acids and other ingredients.

The closest technical solution is the method of clonal propagation under autotrophic conditions in hydroponics, where fructose and mineral elements are used as a nutrient medium by periodically irrigating the bases of the cuttings with a content of regenerants of at least 60 W PAR / m by periodically flooding them with a solution (patent No. 2617948, published 04/28/2017 Bulletin 1, CIM AO1N 4/00).

The disadvantage of the prototype method is the systematic and periodic feeding of regenerants, which complicates the method and increases costs.

The technical result is the simplification of the method, reducing costs.

The technical solution of the claimed object is that the nutrient solution is prepared on the preparation FLORON at a concentration of 0.5% with the addition of potato pectin in the amount of 0.3-0.4% of an aqueous solution, and the pectin is prepared from potato peelings of healthy tubers of the same variety or hybrid propagated regenerates.

The method of implementation is as follows.

The composition of the preparation "FLORON" includes: free amino acids 4%, biostimulating and root-forming factors - 1.46%, cytokinins 0.03%. The total amount of organic matter - 8%, nitrogen - 1%. phosphorus - 10%, potassium - 10%, boron - 0.25%, molybdenum - 0.20%.

Potato pectin, prepared from healthy tubers of the same variety, is added to the FLORON water solution. Potato peelings are crushed to 1 * 1 mm, the hydrolysis of pectin extraction is dissolved in citric acid at pH -1 and the optimum temperature is 70-100 ° C and the hydrolysis time is 2-5 hours, the precipitation of pectic substances is up to 96% ethyl alcohol (the ratio of solution to alcohol is 1: 1.5), the separation of the precipitated pectin by centrifugation and drying in a spray dryer. The content of pectic substances in the peel of potatoes is on average 14.1%.

Pectin in this concentration (0.3-0.4%) is well soluble in an aqueous solution of FLORON.

Pectin is the best organic orderly of regenerants, capable of removing harmful trace elements and natural elements from the tissues of young plants, poisons such as pesticides, radioactive elements, heavy metals, etc.

With the introduction of the solution FLORON pectin normalizes metabolism.

The amount of the solution is 0.3-0.4%, mixed with a 0.5% solution of FLORON provides sufficient nutrients for the normal development of seedlings. It was established experimentally that during the ontogenesis of regenerants it is reduced by several days.

One of the main factors in tuberization in an aero-hydroponic plant is the onset of formation of stolons and tubers. This phase of development entails the formation of tubers and, bringing them to standard sizes can be removed and stored for storage. Hydroairpower installation allows you to harvest minitubby several times. In this regard, the purpose of our research was to bring to standard sizes of seed tubers, and to enable the growth of small and newly formed.

As a result of the study, it was revealed that the treatment of plants with in vitro stimulants and their mixture with pectin significantly influenced the onset of stolon and tuber formation. Since the treatment with the known biological preparation Baikal-EM (contourl) practically did not affect the beginning of this phase, and the processing with pectin somewhat restrained, which cannot be said of FLORON and its mixture with pectin.

Processing FLORON accelerated the beginning of the formation of stolons and tubers by 50%, and the combined use of FLORON 0.5+ Pectin 0.3-0.4% reduced it by 50%. Consequently, the Red Scarlet variety, when treating plants with pectin, began to form tubers for 25 days, Baikal-EM-21 day, FLORON 12 day, and the combined use of FLORON 0.5% + pectin 0.3-0.4% aqueous solution on 6- Oh and Baikal-EM + pectin on day 19. In the same proportions marked the beginning of the formation of tubers in all other varieties.

Consequently, the onset of tuber formation was significantly affected by the treatment with FLORON and FLORON + pectin in the proposed concentrations and some difference was noted for varietal characteristics.

Treatment with drugs and their mixtures also had a significant impact on the formation of the total number of healthy seed minitubes.

Plants treated with a mixture of FLORON and pectin formed the maximum figures for plant height, number of stems and leaf apparatus, which contributed to an increase in photosynthetic activity. In this variant, 28-38 units / plant of standard tubers were formed in the section of varieties. In the variant with mixture treatment (FLORON + pectin), the number of tubers doubled. The maximum number of standard, virus-free seed tubers was formed on hybrid 10.3 / 228 70 pcs. / 1 plant.

The obtained seed material is resistant to a complex of diseases due to the introduction of pectin prepared from healthy tubers into a solution, and can be used for further breeding purposes. This reduces the cost of various expensive ingredients, simplifies the method of propagation of hybrid plants.

Claims (1)

A method of propagation of immunostable potato samples in vitro on aerohydroponics, including irrigation of the bases of the cuttings of the regenerants, characterized in that the water preparation Floron is administered in a concentration of 0.5% with the addition of pectin in a concentration of 0.3-0.4% of an aqueous solution of Floron , moreover, pectin is prepared from potato peelings of healthy tubers of the same variety or a hybrid of propagated regenerants.