RU2626722C2 - Method and device for fighting against botrytis cinerea while growing grafted grape seedlings - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: method comprises the pre-treatment: blinding stock eyes, preparing graft eyes, inoculation, stratification, treatment of the inoculations against gray rot. Wherein the treatment in the stratification period is carried out with steam containing 0.2% of Albit solution at the temperature of 20-25°C and the air humidity of 90-95%, until the stock eye is developed on 1.5-2 cm. Then the steam temperature is raised to 45-50°C and maintained for 10 minutes. The device comprises a control unit, containers with a substrate, a heating element and a substrate temperature sensor. Wherein the device is made in the form of a film micro-greenhouse with a heating element placed on the bottom under a layer of substrate. In addition, the device comprises two steam generators installed at the right and left ends of the micro-greenhouse and equipped with steam temperature and humidity sensors and a timer. Steam is supplied from the steam generators by means of a pipe with openings and steam reflectors located on the ridge of the micro-greenhouse.

EFFECT: inventions allow increasing the yield of grafted seedlings due to destroying Botrytis cinerea while maintaining the natural balance of beneficial microorganisms and observing biological norms for human protection.

2 cl, 1 dwg, 2 tbl

Description

The invention relates to the field of agriculture and may be applicable in crop production, horticulture and viticulture for growing plant cuttings.

The main way to combat infectious plant diseases is traditionally the use of chemicals. The use of pesticides in large volumes caused problems of resistance of harmful objects, as well as economic - the need for additional costs for protective measures and environmental - the safety of the environment.

The disadvantages of using fungicides, insecticides and similar artificial preparations are the deterioration of the environmental situation, the violation of the natural balance, the increased content of harmful substances in agricultural products. In this regard, in recent years, intensive development of alternative methods of protecting plants from infectious diseases is ongoing. One of these methods is the use of biological agents. With a relatively high antimicrobial activity, they practically do not upset the biological balance, while the quality of plant products improves significantly.

Numerous attempts have been made to use antagonist microbes in crop production. It is proved that the use of biological methods is economically more efficient than traditional chemical methods. (Plant Protection and Quarantine. No. 2 - 1997. P. 13. The Second All-Russian Scientific and Theoretical Conference "Progressive Environmentally Safe Technologies for Storage and Complex Processing of Agricultural Products to Create Foods of Increased Nutritional and Biological Value", Part II, October 1-4 1996).

However, despite the obvious preference for biological measures, they have received limited use, primarily due to the lack of active producer strains, proven technologies for the production of biological products and reliable methods for their use in crop production.

In recent years, drugs based on bacteria from the Pseudomonas group have begun to appear in Russia. A known drug used for preplant treatment of cuttings, under the commercial name Rizoplan, whose active principle is living cells of the bacteria Pseudomonas sp., Selected for their ability to suppress phytopathogens - root rot pathogens. Rizoplan protects plants from damage by pathogens of late blight, downy mildew, bacteriosis (Plant Protection and Quarantine. No. 8, 1996 - P. 14-15).

However, the drug has the following disadvantages: firstly, the cultivation of bacteria Pseudomonas sp. VKPM B-3481 "is produced on rich nutrient media (glycerin, peptone, King B medium), which are scarce and expensive, and secondly, due to the fact that the bacteria Pseudomonas are non-spore bacteria, the treatment of seeds with Rizoplan with subsequent drying leads almost to the complete death of bacteria within 2-3 days.This makes it impossible to treat the seeds with Rizoplan in advance, which creates great inconvenience when working with this converter.

Known biological drug Pseudobacterin-2 protective and stimulating action based on cells of bacteria of the genus Pseudomonas aureofaciens. The active principle of the drug is live bacterial cells that colonize the roots, stems and leaves of plants, while inhibiting the growth and development of pathogens. The preparation almost completely eliminates root rot from the plant; the increase in the yield of cucumber and tomatoes in greenhouses when using Pseudobacterin-2 was 18-20% (Plant Protection and Quarantine No. 7, - 1999. - P. 14-15).

The disadvantage of this drug, as in the first case, is the limited lifespan of bacterial cells, which is highly undesirable for an agricultural producer.

The effectiveness and profitability of biological products can be improved if used in combination with plant growth regulators. Recently, biological preparations of complex action have been created, combining both fungicidal and stimulating signs. This technique directedly affects the growth and development of plants from seedlings to harvest, activates their immunity. Growth regulators, as a rule, have a multifunctional effect on plants - they increase their height, weight of the aerial parts and roots, productive bushiness, contribute to the formation of larger grains, increase the resistance of plants to adverse environmental factors and diseases.

But all these drugs can not be successfully used in viticulture in the fight against pests and diseases. For example, Botrytis cinere (gray rot) is an infectious disease. This micro-fungus develops well on dead parts of plants and is considered a saprophyte. Gray rot is spread by conidia. Conidia, even on the mother liquor, penetrate into the tissue of shoot nodes through the antenna, leaf or stalk of the bunch and lead to the death of tissues. On plantations of a vineyard or on a mother liquor in rainy cool weather, the number of conidia in the air increases and, as a rule, their billionth mass, "smoking" and spraying, is released from the surface of sick clusters. Gray rot is dangerous because an outbreak of its development takes place during the ripening of grapes, when the use of chemicals in the vineyards is not allowed; can settle on the surface of the eye scale, at the base of it and on the wounds remaining after the leaves fall. When harvesting cuttings in November-December, especially in mild and rainy weather, hyphae of gray rot penetrate deeply into living tissue, as a result of which the main and replacement buds die during storage of cuttings, spots appear on the bark of shoots with dead tissues. It often happens that from a healthy-looking eye in the process of stratification of grafted cuttings, a shoot developing from a kidney is affected by gray rot. It follows from the above that conidia must be destroyed by chemical treatment in the vineyards, that is, before they are stored.

There are various methods of chemical processing of cuttings. Currently, chemical methods for controlling gray rot have spread, one of which is selected as a prototype. (Grape nursery - Maltabar L.M., Kazachenko D.M. - Krasnodar 2009, p. 117-121).

Before harvesting cuttings, abundant spraying of uterine plantations with 2.4-2.8 kg / ha of mycodiphol or 3.6-4.2 kg / ha of orthofoltan is carried out. Before laying for winter storage, harvested cuttings are subjected to chemical processing no later than 24-48 hours after cutting.

In Germany, chinosol (Chinosol W), consisting of 67% 8-hydroxyquinoline sulfate + 30% potassium sulfate, is used to treat cuttings.

In Hungary, Solvokhin Extra (Chinoin, Budapest) 70% 8-hydroxychigoline, 14% potassium sulfate, 15% ethylenedialine-tetraacetic acid, 1% neutral fatty sulfate are used.

For disinfection, the bundles of rootstock and graft cultivars are completely soaked in a 0.5% quinosol solution. The duration of soaking depends on the temperature of the water: at 5 ° C it lasts 5 hours, 10 ° C - 3 hours, 20 ° C - 2 hours. Soaking cuttings in quinosol is carried out after soaking them in water. Excessive concentration of disinfectant inside the cuttings and on their surface negatively affects the formation of callus and roots, the development of the kidneys and dramatically reduces the yield of seedlings.

All of these chemical control measures are ineffective and dangerous to the health of workers involved in the cultivation of seedlings. The currently used set of measures, the agrochemical chemical methods for combating gray rot cannot solve the problem of protection against gray rot, especially when growing grape seedlings.

In addition, the issues of stratification techniques and maintaining an optimal automatic regimen for vaccine stratification in domestic and foreign practice are still very poorly developed.

The known technology of open stratification in an environment of intensely moistened air (Mishurenko A.G. Grape nursery. - 3rd ed. - Moscow, 1997). Stratification chambers are usually heated with water heating, and the necessary air humidity in the chambers is maintained by evaporating water from special bathtubs. To further moisturize and remove mold on grafted cuttings, they are periodically doused with water from special nozzles, and also ventilated with special fans. Grafted cuttings coated with antiperspirants are placed in trays (baskets). The metal basins of the trays have several holes for draining water. The stratification chamber (room) has a capacity of up to 200 thousand grafted cuttings. In the vaccination complex there are from 6 to 10 such chambers. In the chamber in 2 rows metal racks are placed. Racks are three-tier. Between the racks there is a metal track. Trays with grafted cuttings are mounted on a trolley and transported to stratification chambers. They are stacked on racks, starting from the lower tier. The camera is unloaded in the reverse order. The camera is loaded for no more than 2 days. In each stratification chamber, there are two types of heat exchangers: water radiators and water baths located above the racks. Using radiators, the air in the chamber is heated to 28-29 ° C, and with the help of coils, which are lowered into the water evaporators, the water temperature in the baths reaches 35-36 ° C. Water heated to this temperature evaporates and saturates the air in the chamber in vapors, which helps to maintain high (up to 100%) air humidity in the chamber. However, with intensive ventilation and the movement of vapors on the upper racks, the tops of the grafted cuttings dry out. To prevent this, additionally moisturize by spraying with water from the nozzles. In the first 6-7 days, the temperature in the chambers is maintained at 29-30 ° С, air humidity is 97-100%. On the eighth day, the temperature in the chamber is reduced to 26 ° C and maintained under such conditions until the end of stratification. The privatny part (the tops of the grafted cuttings) must be on the same level, for which a line is drawn on the inner walls of the box or a movable bar is inserted, to which the tops of the scion should reach. The described installation allows only stratification of vaccinations, while some of the vaccinations are subjected to mold and eye aging. When sawdust dries, their growth and death cease. Stratified vaccinations with the formed root system have to be planted in covers or in an open ground school, while the root system of the vaccinations is violated, and the growth of new roots requires additional nutrients that have already been used up earlier and therefore the survival rate of such vaccinations is sharply reduced. During stratification, it is necessary to create lighting conditions and good air access. When using installations of this type at the end of stratification, when the scion eyes begin to germinate, in poor lighting conditions the shoots quickly stretch and a large amount of nutrients is consumed, depleting vaccinations. The grafted cuttings landed in the school have a low survival rate. The described device is cumbersome, requires a large consumption of electricity and its use does not allow the fight against diseases and pests, as well as other operations from hardening to planting seedlings in open ground. (Grape nursery - Maltabar L.M., Kazachenko D.M. - Krasnodar 2009, p. 140-157).

The ESU-2 M serial electrostratification plant (http://www.ozelenenie-mo.ru/bio/120.php) is known for local heating of grafts in grafts in standard size stratification boxes in order to accelerate the process of fusion of scion with stock and considered in as a prototype. The installation consists of a three-phase step-down transformer TS-2.5 with a primary voltage of 380/220 and a secondary voltage of 36 volts, a shield of the power switch, a temperature sensor, which is a TPK electrical contact thermometer, power cables, 72 heating elements and a control panel. The heating elements are wire frames, on which a heating cable POSHP-1.1 6.4 m long is wound in the form of three main and one additional bypass coil. All heating elements are divided into three groups of 24 elements, each equipped with an ammeter for monitoring the load and individual automatic load switches in the event of a short circuit.

The main disadvantage of this installation is that it cannot provide optimal conditions for the development and emergence of healthy seedlings in the automatic mode provided for by the method proposed above. In addition, it does not seem possible to monitor the passage of the stratification process of vaccinations after treatment with pesticides. When using plants of this type at the end of stratification, when the scion eyes begin to sprout, in poor lighting conditions the shoots quickly stretch, and a large amount of nutrients is consumed, depleting vaccinations. At the same time, to maintain a high (up to 100%) air humidity in the chamber, a large energy consumption is required, since the stratification chamber is a huge room that needs to be fully heated. The installation does not provide for the possibility of creating different temperature conditions at the tops and bases of the same inoculations, which dramatically reduces the yield of inoculations with circular callus.

The objective of the present invention is to develop a method and device that allows to increase the yield of grafted seedlings by destroying Botrytis cinere while maintaining the natural balance of beneficial microorganisms and observing biological standards for human protection by treating vaccinations with steam at elevated temperatures while saturating macro and microelements with cuttings.

The problem is solved using the proposed method of combating Botrytis cinere in the cultivation of grafted grape seedlings, including pre-treatment: blinding the rootstock eyes, preparing graft eyes, vaccination, stratification, vaccination treatment during the stratification period with steam containing 0.2% Albit solution a temperature of 20-25 ° C and an air humidity of 90-95%, and a further increase in steam temperature to 45-50 ° C after the development of the rootstock eye by 1.5-2 cm and keeping it for 10 minutes and a device for its implementation. A device comprising a control unit, containers filled with a substrate, a heating element, a substrate temperature sensor, is made in the form of a film microheat with heating elements placed on the bottom under the substrate layer, with two steam generators installed on the right and left ends of the microheat, equipped with steam temperature sensors humidity and timers, the steam supply from which is carried out by means of a pipe with holes and steam reflectors located on the ridge of the micro-greenhouse.

The novelty of the claimed method lies in the fact that to provoke the development of the fungus, steam is used with a temperature of 20-25 ° C and an air humidity of 90-95%. An indicator of the complete germination of Botrytis cineree conidia is the formation of a green cone from a rootstock eye 1.5–2 cm high. When the cone reaches a predetermined height, the steam temperature is gradually and continuously increased to 45–50 ° C, and such a high temperature is maintained for 10 minutes critical boundary, after which comes the death of gray rot. The rootstock cuttings are first treated in a micro-greenhouse with an optimum temperature of 20-25 ° C, which creates comfortable conditions for saturation with moisture and germination of Botrytis cinerea conidia. The thermal conductivity of the cuttings is very low in low-moist cuttings (average water content of 45-50%) and the pores are filled with air. If the cuttings are moistened, then the pores are filled with water, taking into account that the thermal conductivity of water is 20 times higher than that of air. The cell membrane is a non-indifferent structural element of plant tissue and the components of cell membranes play an important physiological role in the metabolism of plants. Lignified cell membranes, impregnated with lignin and other inlaid substances, under the action of steam with Albit undergo the reverse process of lignification, which is a normal physiological process associated with the vital activity of lignified cells. Therefore, under the influence of steam, conditions of increased tissue permeability are created, the therapeutic effect of steam and the chemical effect of Albit is enhanced. Under the action of steam, the elimination and neutralization of harmful substances, microbes, fungi is accelerated, and the regenerative process of cuttings improves. This technique provides not only the death of gray rot with minimal labor and money, but also an increase in the yield of standard seedlings, their survival on plantations, which ultimately accelerates the laying of new vineyards.

The novelty of the claimed device lies in the fact that it allows, in accordance with the proposed method, to ensure the maintenance of optimal conditions automatically and at the same time as carrying out stratification, to control pests and diseases of grapes. Using the proposed device is regulated depending on the phases of development of seedlings of the temperature level of mineral nutrition, pest and disease control during stratification. In addition, the device provides the ability to create different temperature conditions at the tops and bases of the same grafts due to the temperature difference of the root layer (substrate) provided by the heating element and air by steam generators. One of the steam generators is used for disinfection, and the other provides plant nutrition. Steam is supplied by means of a pipe with holes and steam reflectors, which are cone-shaped nozzles for the directed action of steam.

Important in the proposed method is the fact that in order to accelerate the reproduction, healing of plants from phytopathogenic infection, saturation of rootstock cuttings with nutrients for better callus formation, the composition of the vapor is 0.2% Albit solution. Albite contains the purified active substance poly- and beta-hydroxybutyric acid from soil bacteria Bacillus megaterium and Pseudomonas aureofaciens. In natural conditions, these bacteria live on the roots of plants, stimulate their growth, protect against diseases and adverse environmental conditions.The composition of the drug also includes substances that enhance the effect of the main active substance, a balanced starting set of macro- and microelements (N, P, R , Mg, S, Fe, Mn, Mo, Cu, Co, B, I, Se, Na, Ni, Zn) and terpene acids of coniferous extract. "Albit" does not contain living microorganisms (but only the active substances from them), which makes the drug more stable, less susceptible to environmental influences. "Albit" is characterized by low cost and environmental friendliness of biological products, but at the same time, in terms of efficiency and stability, it approaches chemical ones.

In FIG. 1 shows a General view of the device, where 1 is a control unit; 2 - capacity with a substrate; 3 - heating element; 4 - substrate temperature sensor; 5 - steam generator; 6 - steam temperature sensor; 7 - steam humidity sensor; 8 - timer; 9 - pipe for supplying steam.

The claimed method is as follows.

The invention was tested in 2015 before the start of the vaccination campaign. The shoots of the rootstock were removed from the storehouse and in parts transferred to the preparatory room of the vaccination complex, but not earlier than 10-15 days. The preparation of the stock consisted of a thorough inspection and culling of all insufficiently matured and non-standard in thickness vines. Then the vines were cut, all eyes except the top were carefully removed (blinded), sorted by thickness, soaked for 2-5 days and put on stratification.

A special installation has been developed that provides optimal humidity and temperature conditions for air and substrate, ranging from 25 to 90 ° С. During stratification, the installation automatically maintains the air temperature in the micro-greenhouse 20-25 ° C, the air humidity in the range 45-100% due to the supply of steam through the tubes directly into the film micro-greenhouses.

The thermal elements installed in the micro-greenhouse ensure the maintenance of a constant temperature of the soil due to the cycles of supply and removal of voltage to the fuel elements - a resistive heating cable, which is installed in the substrate layer under the root system of plants. Temperature control is carried out using an electronic thermostat, which measures the temperature using a sensor installed in a heated substrate layer. The heating system is powered by an alternating voltage of 220 V. The device is equipped with a circuit breaker to relieve voltage in the event of a short circuit in the electrical circuit, if the heating cable is damaged and serves as protection against electrical shock to personnel.

When the supply voltage is supplied to the control unit 1 and the set temperature controlled by the temperature sensor of the substrate 4, the substrate layer is heated using the heating element 3. When the substrate temperature is equal to the set value on the control unit 1, the heating is automatically turned off. Heating time depends on the ambient temperature.

The operation of the steam generators 5 is carried out using the control panel located in the steam generators themselves. Both steam generators are equipped with steam temperature sensors 6, humidity 7 and timers 8. On the control panel set the necessary parameters for temperature, humidity and exposure time. One of the steam generators serves to disinfect planting material from pests and diseases, and to maintain humidity of the air and substrate in the micro-greenhouse. The second - provides nutrition to plants: they dissolve fertilizers at a temperature of 35-40 ° C and serve along with steam, which reduces the consumption of drugs and increases the efficiency of their use. The supply of steam from the steam generators is carried out by means of a pipe 9 with holes and reflectors of steam, which is located on the ridge of the micro-greenhouse.

Under the action of steam, the elimination and neutralization of harmful substances, microbes, fungi is accelerated, which improves the regenerative process of cuttings. The saturation of cells with a therapeutic agent occurs until a green cone, an unopened green leaf 1.5-3 cm tall, develops from the upper eye. The formation of a green cone with a height of 1.5-2 cm is an indicator that the conidia of the fungus have sprouted and it is possible to raise the steam temperature to 45-50 ° C, and hold for 10 minutes. At this time, all physiological processes in the cuttings are most active, cambium and other elements of the shoot structure are very active, which are directly related to the processes of growth and root formation. During this period, the restoration of the conductive system of blood vessels, the restoration of the movement of nutrients to the green shoot is noted. And only after all the processes in the cuttings are activated, the top of the cuttings together with the green cone is removed from the rootstock, cutting off below the line of leaf scars, and a healthy scion graft with a swollen eye is grafted into this place. The connection zone contains the largest number of tissues and blood vessels, the most active during intergrowth. To the place of vaccination is supplied apiary and physiologically active substances (auxins), which stimulate tissue regeneration and provide better fusion of the components of the vaccine. Using rootstock and graft with vessels already working, and ensuring, as it were, a natural continuation of secondary growth vessels from the graft to rootstock, the most favorable conditions are created for fast and high-quality intergrowth. Vaccinations are carried out in any conventional way, put on stratification for 9-10 days, and then planted in covers. A prerequisite for this is the presence of drip moisture. The bound cuttings are placed in the installation we created at a temperature of 20-25 ° C, air humidity of 85-90% until a green cone develops from the upper eye. The conidia of the fungus germinate, after which the steam temperature is raised to 45-50 ° C and held for 10 minutes, which leads to the death of gray rot, while maintaining the viability of the cuttings. For a more accurate assessment of the degree of death of gray rot, a batch of 200-300 pieces of cuttings is taken, covered with a wet burlap that retains moisture, and kept at a temperature of 20-25 ° C for 4-5 days, after which the question of the need for re-processing is decided.

In the presence of gray patches on transverse sections and browned tissues with longitudinal sections, more than 0.2% affected by gray rot, the treatment can be repeated. Parties of cuttings are allowed to be vaccinated, if not more than 0.2% are affected by gray rot. After that, they are planted in covers by sticking into a moist sterile substrate. The grafted cuttings are planted to a depth of 8-10 cm. To ensure the required planting depth, grafts need to be taken at the bottom so that the fingers serve as a limiter, preventing a deeper planting. After this, the substrate is sprinkled with sand in order to fill the voids, and then abundant watering is carried out. The moisture content of the substrate is maintained by systematic irrigation with warm water at the level of 85-90% of the PPV. The temperature of the air and the substrate should be at a level of 20-25 ° C, although in sunny weather it rises significantly, which requires systematic ventilation. After the appearance of the roots, 2-3 times are fed with a hydroponic solution. During the growing period, seedlings are removed and, if required, the plants are treated against mildew. Subject to all conditions, the roots of the cuttings are formed on the 8-10th day. It has been experimentally established that it is most advisable to grow vegetative seedlings for 60-65 days. The grafted vegetative seedlings should have a circular growth of components, hardened growth of more than 12 cm and a lump of substrate well penetrated by the roots. Vegetable seedlings should be well hardened before planting in open ground; for this, the film is raised on a micro-greenhouse, and after 5 days they are removed, and the seedlings are planted in open ground.

The experimental data obtained as a result of the experiment are presented in Table. 12.

In Tab. Figure 1 shows the effect of high steam temperature on the death of conidia of gray rot and the yield of seedlings using the example of Crystal Rootstock Kober 5 BB.

As can be seen from Table. 1, when the rootstock cuttings were processed at t 60 ° C for one hour, the conductive tissues of the rootstock cuttings were damaged and the yield of grafted seedlings Crystal variety was only 8.8%. Treatment with the same temperature for 30 minutes also led to burns of the cuttings tissue with steam, but the yield of grafted seedlings increased significantly and amounted to 15%. Three-time spraying with a 0.1% quinosole solution turned out to be ineffective - a large number of vaccinations were affected by gray rot, and the yield was only 27.4%, while in the proposed method, the yield of vegetative seedlings was 80.6%.

In Tab. Figure 2 shows the effect of the Albit preparation on the yield of vaccines with circular callus growth and development of vegetative seedlings using the example of the Pamyaty Smirnov variety, Kober rootstock 5 BB.

The experimental data obtained as a result of the experiment indicate that with the use of "Albit" the best callus formation in vaccinations, the most intensive growth of shoots was noted. The sheet surface area in the second embodiment is 75.6 cm ² , and according to the technology used in production, the control is 20.7 less. At the same time, the growth of shoots was enhanced both in the apical and in the lateral meristem; in general, the quality of seedlings increased.

Experimental data confirm that the proposed method and device can increase the yield of grafted seedlings by destroying Botrytis cinere by treating steam vaccinations with elevated temperature and simultaneously saturating the cuttings with macro and microelements, while maintaining the natural balance of beneficial microorganisms and observing biological standards for human protection.

Claims (2)

1. A method of controlling Botrytis cinerea when growing grafted grape seedlings, including pretreatment: blinding the rootstock eyes, preparing scion eyes, grafting, stratification, treatment of gray rot vaccinations, characterized in that the treatment during the stratification period is performed with steam containing 0.2 % Albit solution at a temperature of 20-25 ° C and an air humidity of 90-95%, until the rootstock eye develops by 1.5-2 cm, then the steam temperature is raised to 45-50 ° C and held for 10 minutes. 2. A device for implementing the method according to claim 1, comprising a control unit, containers with a substrate, a heating element, a substrate temperature sensor, characterized in that the device is made in the form of a film microheat with a heating element placed on the bottom under the substrate layer, further comprises two a steam generator installed on the right and left end of the micro-greenhouse and equipped with steam temperature, humidity and timer sensors, the steam supply from which is carried out by means of a pipe with holes, and steam reflectors, located on the ridge of micro-greenhouses.

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