RU2521992C1 - METHOD OF MICROGRAFTING GRAPES in vitro - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: method comprises micropropagation of test-tube plants and their planting into the liquid nutrient medium with the addition of macronutrients, vitamins and biological products. At that, based on the substrate Murashige-Skoog the activated carbon is eliminated, the content of salts and acids is reduced 2-4 times and Humate+7B is added in an amount of 5-10 ml/l.

EFFECT: method enables to improve the efficiency and accelerate propagation of treated from the viral infection promising grape varieties.

2 tbl, 1 ex

Description

The invention relates to the field of agriculture, in particular to methods of plant propagation, and can be used in viticulture for accelerated propagation of promising grape varieties recovered from a viral infection by reducing the cost of expensive preparations.

A known method of propagation of grapes, in which they isolate meristematic explants, planting and cultivating them (patent No. 2120739 from 10.27.1998, IPC A01H 4/00).

However, in the known technical solution for stimulating microexplants, an electromagnetic microwave field is used in combination with a narrow-band laser beam, which complicates the method and increases costs.

There is also known a method of microclonal propagation, including the separation of test plants on a nutrient medium Murashige-Skoog with the simultaneous use of biological products (patent No. 2265319 from 10.12.2005, IPC A01H 4/00).

The complexity of the known technical solution lies in the fact that biological products of gibberellin and cytokinin breeds are isolated from fungi, which affect the growth and development of plants at the stage of introducing meristems into the culture.

However, explants for their development require additional introduction of other elements for their development, which reduces the effectiveness of the method and increase costs.

The closest technical solution is a method in which micropropagation is carried out in vitro by microcentering of test plants and planting them on a liquid nutrient medium with the addition of macronutrients, vitamins and biological products (patent No. 2264706 from 27.112006, IPC A01H 4/00, A01N 25/00, C12N 5/00).

However, in the prototype method for the propagation of microcherenkov use expensive drugs to form a nutrient medium, which complicates the method, increases the cost of production.

The technical result is a reduction in the cost of production and increasing the efficiency of the method.

The technical solution of the claimed object, in contrast to the known one, consists in the fact that activated carbon is excluded from the composition of the Murashige-Skoog substrate, salts and acids are reduced by 2-4 times, and Humate + 7B is added in an amount of 5-10 ml / l.

The method is as follows.

Humate + 7B is readily soluble in water, easily absorbed by plants, mobilizes its immune system, stimulates the development of a powerful root system, promotes enhanced nutrient intake, intensifies metabolic processes in the plant cell, reduces nitrate content by 2 times, increases the content of chlorophyll, vitamins, sugars and other valuable substances, it stimulates the effect of all trace elements used in mixtures with humate.

Unlike the known types of humates, the proposed Humate + 7B contains 60-65% of humates, trace elements (iron, copper, zinc, manganese, molybdenum, cobalt and boron). In this combination with the advantage of boron (B), humate was not used for microcherenking of grapes; boron is necessary for plants for normal growth and development. Boron functions are associated with metabolism, the transport of sugars through membranes, the synthesis of DNA, RNA and phytohormones, the formation of cell walls and tissue development. Lack of boron in the body causes various diseases. Given the complex of micro- and macrocells used as a nutrient medium, Humate + 7B supplements as valuable substances that combine a joint positive effect on the result of experiments.

Example. Regenerant plants 8-10 cm in size were cut into fragments, which included a node with a leaf and a kidney (the lower part of the internode was 1-1.5 cm longer than the upper one), the obtained microcranes were planted in biological tubes 40 × 120 mm in size on a nutrient medium. The tubes were closed with foil and placed in a culture room with appropriate conditions for the method.

The experience was laid on the Augustin grape variety in 5 variants, Table 1. In each version, 3 repetitions. In each repetition of 6 tubes with microstrings. We studied the effect of modified nutrient media on the growth and development of a grape plant in vitro.

The technology of grafting of test plants is conventional. The date of cuttings was 01/08/2012. Observations of plants were carried out for 41 days until 09/11/2012 - daily, noting the date the roots and leaves appeared. Measurements of plant height, counting of leaves and main roots were carried out 41 days after cuttings.

In all cases, the beginning of root formation was noted on the 8th day after the cuttings. On the 13th day after the cuttings, all plants took root.

It was noted that root growth in option 5 also increased significantly in the number of roots that exceeded control.

Leaf formation in option 5 began on the tenth day after the cuttings, and on day 13, leaf formation began on all variants of the experiment.

Further observations of the growth and development of plants showed that explants in variants 2 and 3 on poor nutrient media, although initially showed good rooting results, lagged far behind growth and development compared to options 1, 4, 5 (Table 2).

Significant differences in the number of roots, leaves, and plant height are observed in variants 4 and 5. Explants placed on these media underwent more intensive growth of the plant in height and root formation compared to the control variant (Table 2).

The best results on the growth and development of plants showed option 5 on a nutrient medium No. 4. Introduction to the composition of nutrient media, in addition to mineral salts, the liquid concentrated organomineral preparation Gumat + 7B, was essential for the growth and development of the explant in vitro.

These results are summarized in table 2, from which it follows that the nutrient medium No. 4 is the most optimal, providing for a relatively short period an increase in the number of main roots, leaves and plant height.

In nutrient media No. 4 and No. 5 (Table 1), the amount of agar agar, sucrose, sodium, magnesium, potassium and calcium salts, mesoinositol, copper sulfate and nickel chloride, nicotinic acid, pyridoxine, thiamine, iron sulfate, trilon is significantly reduced B, activated carbon, due to a significant reduction of these elements in the nutrient medium and the addition of Humate + 7B in an amount of 5-10 ml / l.

Therefore, in the proposed method reduces the cost of the nutrient medium and increases its effectiveness.

Table 1 The composition of the nutrient medium for rooting and growth of test tube grape plants (in vitro method) 07/31/2012 Mg / L, ml / L No. Name of drugs Option 1 Option 2 Option 3 Option 4 Option 5 Control, modified MS environment Culture medium No. 1 Culture medium No. 2 Culture medium No. 3 Culture medium No. 4 one Agar agar 7000 7000 7000 7000 7000 2 Sucrose 15,000 20000 20000 10,000 10,000 3 KNO ₃ - potassium nitrate 950 475 475 four NH ₄ NO ₃ - ammonium nitrate 825 69 69 5 MgSO ₄ × 7H ₂ O - magnesium sulfate 185 185 93 93 6 CaCl ₂ × 2H ₂ O - calcium chloride 220 166 83 83 7 KH ₂ PO ₄ - potassium phosphate 85 34 34 8 Mesoinositis one hundred fifty 25 25 9 KI - Potassium Iodide 0.42 0.42 0.42 10 H ₃ BO ₃ - boric acid 3,1 3,1 3,1 eleven ZnSO ₄ × 2H ₂ O - zinc sulfate 4.3 4.3 4.3 12 MnSO ₄ × 4H ₂ O - manganese sulfate 1,1 1.12 1.12 13 CuSO ₄ × 5H ₂ O - copper sulfate 0,025 0.013 0.013 fourteen NiCl ₂ - Nickel Chloride 0,025 0.013 0.013 fifteen A nicotinic acid one 0.5 0.25 0.25 16 Pyridoxine B ₆ one 0.5 0.1 0.1 17 Thiamine B ₁ one 0.2 0.1 0.1 eighteen FeSO ₄ × 7H ₂ O - ferrous sulfate 27.8 13.9 13.9 19 Trilon B Na ₂ EDTA × 2H ₂ O 37,2 18.6 18.6 twenty Humate + 7B (ml.) 10 10 10 5 21 Activated carbon 5000 pH environment 6.6 6.6 6.6 6.6 6.6 Note: Trilon B previously dissolved in 5 ml bidistilled water.

The influence of various nutrient media on the growth and development of grape plants during microcherenking (in vitro method)

Table 2 Date of registration September 11, 2012, 41 days after the cuttings Options Number of main roots Number of leaves Plant height, cm Repetitions Total Average per 1 plant Repetitions Total Average per 1 plant Repetitions Total Average per 1 plant one 2 3 one 2 3 one 2 3 Option control, modified MS environment 1.7 1,5 2,3 5.5 1.8 0.17 5.50 5.83 17.5 5.8 7.0 5.3 6.3 18.6 6.2 Culture medium No. 1 2.0 1,5 1.8 5.3 1.8 3.17 3.33 1.00 7.5 2,5 2.2 3 1,2 6.4 2.1 Culture medium No. 2 1.7 1.3 1,0 4.0 1.3 0.5 1.00 1.17 2.7 0.9 0.5 0.8 0.7 2.0 0.7 Culture medium No. 3 1.7 1.8 1.8 5.3 1.8 6.33 5.50 6.33 18.2 6.1 7.0 6.5 7.3 20.8 6.9 Culture medium No. 4 1.8 2.0 2.2 6.0 2.0 6.33 6.50 5.83 18.7 6.2 7.3 7.7 7.8 22.8 7.6 HCP-1.25 HCP-2.14 HCP-2.39

Claims (1)

In vitro microcirculation of grapes, including micropropagation of test plants and planting them on a liquid nutrient medium with the addition of macronutrients, vitamins and biological products, characterized in that activated carbon is eliminated on the basis of the Murashige-Skoog substrate, salts and acids are reduced 2-4 times, and add Humate + 7B in an amount of 5-10 ml / l.