RU2467556C1 - Method of biological control of alternaria of grape - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to the field of agriculture, in particular, to viticulture. In the process against the pathogen Alternaria tenuissima (Kunze ex Pers.) Wiltshire the biofungicide based on antagonist fungus Trichoderma viride Pers. ex S.F. Gray strain 4097 is used, the titer is at least 1.0×10⁹ CFU/ml. The consumption rate of biofungicide is 1.5-3.0 l/ha. Processing is carried out by blocks: the first is carried out prior to harvesting to suppress the current infection, and the second is after harvesting to reduce the accumulation of infection.

EFFECT: method enables to increase efficiency and environmental safety of control of Alternaria, to reduce phytotoxic effect of treatments to maintain the plants immunity through the use of biological defense technology.

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Description

The invention relates to agriculture, in particular to the biological protection of plants from diseases.

There is a known chemical method of combating grape alternariosis with the help of Bordeaux liquid (Pests, diseases and weeds in vineyards / Zh.A. Chichinadze, N.A. Yakushina, A.S. Skorikov, E.P. Stranishevskaya. - Kiev: Agrarian science, 1995. - S. 79-80).

The disadvantage of this method is the negative effect of Bordeaux liquid on the physiological state of grape plants, which is expressed in the drying of the leaf surface and is especially negatively manifested in dry periods of summer, the frequency and duration of which has increased in recent years in southern Russia, the main viticulture zone. The direct antifungal effect of Bordeaux fluid on the causative agent of alternaria grape (Alternaria tenuissima (Kunze ex Pers.) Wiltshire (syn. Helminthosporium tenuissimum Kunze, Macrosporium tenuissimum Fr. (Ellis, 1971,477) - identification by F. B. Hannibal, VIZR, St. Petersburg. according to our observations, it is noted that alternariosis affects grapevines with a weakened immune system more severely, and the drying effect of this treatment reduces the plant's immunity. As a result, the use of Bordeaux fluid in the fight against grape alternariosis is absolutely not effective.

A known biological method of combating potato alternariosis is the pathogen Alternaria solani, using a biocomplex, which includes the microbiofungicide Phytosporin-M based on Bacillus subtilis, strain 26D, the anti-stress preparation gumi-20 and organic biofertilizer borogum, g. Biological products for protecting potatoes from diseases / L.I. Pusenkova, V.M. Grez, V.N. Zeyruk, M.K. Derevyagina, I.V. Maksimov. - Protection and quarantine of plants, No. 10, 2010. - P.26-28. Microbiofungicide Fitosporin-M and several other biological products based on the same bacterium (other strains) are registered in the “List ...” (List of pesticides and agrochemicals approved for use in the Russian Federation in 2010. - Moscow, 2010. - C.122 , 134) against alternariosis of tomato and potato. The biological effectiveness of this method is at the level of 52%, which is unsatisfactory for harmful diseases, i.e. of economic importance, which is the alternariosis of grapes. Over the past 5 years, this disease has developed in the form of explosive epiphytotia on grape varieties - interspecific hybrids in the region of the Western Ciscaucasia. The yield loss from one bush with an average and severe defeat reaches 27.8-38.9%, the quality of grapes deteriorates due to a decrease in sugar by 6.1-9.1% (statistically significant values) (table 1).

Table 1 The influence of grape alternariosis on the crop and product quality, grade Bianka, LLC Yuzhnaya AF, 2009 Degree of disease R, % R% Harvest from the bush, kg Reliable damage,% Mass content of sugars, g / 100 cm ³ Reliable damage,% Weak 40 12.3 9.3 - 19,4 - Average 72 38,0 6.5 27.8 18.6 6.1 Strong one hundred 67.5 5.5 38.9 18.0 9.1 Without defeat 0 0 9.0 - 19.8 - NDS 05 2.14 0.97 Note: P - spread of the disease R - the development of the disease

According to our studies, all biological preparations registered against alternaria in other cultures based on various strains of the bacterium Bacillus subtilis in the “List of pesticides and agrochemicals approved for use in the Russian Federation” until 2010, inclusive, do not show antifungal activity in the fight against Alternaria grape Alternaria tenuissima .

There are known attempts to develop biological methods of combating plant pathogens using various species of mycophilic fungi of the genus Trichoderma.

For example, in soybeans, T. lignorum (Tode) Harz., Strain T-3, and T. koningii Oudem, strain T-10 were used in the fight against root rot. Despite the positive effect on the germination of soybean seeds, the biological effectiveness of these fungi in suppressing diseases was low, at the level of 18.4% (Role of mushrooms Trichoderma spp. In limiting root rot of soybean / E.N. Shendrik. - Immunopathology, Allergology, Infectology, 2010, No. 1. - S.139-140).

There is evidence of antagonistic activity when Trichoderma asperellum MG-97 is used (RF patent No. 2171580) in the biocontrol of phytopathogens of coniferous seedlings, where its biological effectiveness is fixed at 36-42% (Using a new strain of Trichoderma fungi in the biological control of phytopathogens of coniferous seedlings in Middle Siberia / T.I.Gromovykh, A.L. Malinovsky, Yu.A. Litovka, E.A. Shilkina // Collection of Modern Mycology in Russia, Abstracts (1 Congress of Mycologists of Russia) - Moscow, 2002. - S.227-228).

But all of them have an efficiency of up to 50%, which is not enough to control harmful diseases in perennial crops, in addition, alternarioses are not included in the spectrum of controlled diseases in these methods.

There is a report on the biological product Trichodermin based on the fungus T. lignorum, which refers to the activity of this fungus against Alternaria, Phoma, Botritis, etc., recommendations are given on its use in grapes (Internet - http // agrochimiya.ucoz.ua). But the effectiveness of its use is not confirmed by either laboratory or field tests. Meanwhile, this fungus does not show antifungal activity against the listed pathogens of grape diseases, including Alternaria tenuissima, the causative agent of grape alternariosis.

A known biological method of controlling the use of the biological product Trichodermin nova based on T. viride, strain 4097, titer 10 ⁶ CFU / g, manufactured by Bioformatek LLC for the fight against mold mold and root rot (helminthosporiasis, fusariosis) on spring wheat and barley using presowing treatment of seeds and spraying during tillering-earing (List of pesticides and agrochemicals allowed for use in the Russian Federation in 2009. - Moscow: Agrorus Publishing House, 2009. - P.116-117).

But in the spectrum of the antifungal action of this biocontrol agent, causative agents of alternariosis, in particular alternaria grape Alternaria tenuissima (Kunze ex Pers.) Wiltshire, were not noted.

In addition, in all these methods, the features of the use of biological agents in perennial crops (agrocenoses with the continued existence of the same plants, fungi, bacteria, etc.) in one place are not taken into account, i.e. having a more complex structure and interaction mechanisms, as well as having the ability to accumulate infection. In such cenoses, the usual schemes for the use of biological agents, i.e. only to suppress current infection, not effective enough.

The essence of the invention lies in the fact that for the first time to reduce the harmfulness of alternaria grapes - pathogen Alternaria tenuissima (Kunze ex Pers.) Wiltshire, a biological product based on the fungus Trichoderma viride Pers ex SF Gray, strain 4097, with a titer of at least 1.0 × 10 ⁹ CFU / ml (RF patent No. 2380906; List of pesticides and agrochemicals permitted for use on the territory of the Russian Federation in 2009. - Moscow: Agrorus Publishing House, 2009. - P.116-117). Spraying is carried out during the growing season of grapes and consists of 2 blocks. In the first block, treatments are carried out starting with the appearance of the first single signs of the disease and are repeated at intervals of 7-12 days depending on the intensity of the development of the disease, the last treatment in this block is carried out no later than 15-20 days before harvesting, due to the specifics of winemaking. The consumption rate of a biological product is 1.5-2 l / ha, pouring a working solution of 300-1000 l / ha, depending on the processing time. In the second block, 1-2 treatments with the same biological product based on the fungus Trichoderma viride Pers ex SF Gray, strain 4097, with a titer of at least 1.0 × 10 ⁹ CFU / ml, starting 5-15 days after harvesting, are carried out. The consumption rate of 1.5-3.0 l / ha, pouring a working solution of 1000 l / ha. The second treatment in this block is carried out under the condition of a large number of alternariosis infections and depending on the weather conditions of the post-harvest period.

The technical result of the invention is to increase the efficiency and environmental safety of the method of controlling alternaria grapes, as well as reducing the phytotoxic effect of treatments to preserve plant immunity through the use of a biological protective agent, a microbiofungicide based on the Trichoderma viride antagonist fungus, which produces a wide range of metabolites with antifungal action and exhibits high biological efficacy in the fight against grape alternariosis - pathogen Alternaria tenuissima (Kunze ex Pers.) Wilt shire.

Example 1. According to the results of phytosanitary monitoring of vineyards in May, in the phenophase “leaf buds - two leaves opened”, sections of the early hybrids Bianca and Augustin were revealed, where the first manifestation of alternariosis in the form of single dark brown spots of 1-2 mm in size was noted. Biological product (liquid) based on the Trichoderma viride antagonist fungus, with a titer of at least 1.0 × 10 ⁹ CFU / ml, at a rate of consumption of 2.0 l / ha, pre-dissolved in a small amount of water (10 l), allowed to stand for a while . Then it is mixed with water in the tank immediately before spraying and the vineyards are sprayed with the prepared tank mixture using the equipment available at the farm; the pouring rate during this period is 300-350 liters per 1 ha. In the future, according to the results of phytosanitary examinations of these vineyards with an interval of 7-12 days, depending on the intensity of the development of the disease, biofungicide treatments are carried out at a flow rate of 2.0-2.5 l / ha. The last treatment is carried out on 07/30/02/08 15-20 days before harvesting. In early September, these areas are treated with the same biological product at a consumption rate of 3.0 l / ha.

Example 2. According to the results of phytosanitary monitoring of vineyards in July, in the phenophase “fruit production — pea-sized berries, clusters begin to sag”, the plots of late hybrid varieties Moldova, Pervenets Magaracha were revealed, where the manifestation of alternariosis on the leaves in the form of single dark brown spots of size 2 -3 mm. A biological product based on the Trichoderma viride antagonist fungus (liquid) at a flow rate of 2.0-2.5 L / ha is pre-dissolved in a small amount of water (10 L), allowed to stand for a while. Then it is mixed with water in the tank immediately before spraying and the vineyards are sprayed with the prepared tank mixture using the equipment available at the farm; the pouring rate during this period is 700-750 liters per 1 ha. In the future, according to the results of phytosanitary examinations of these vineyards with an interval of 7-12 days, depending on the intensity of the development of the disease, biofungicide treatments are carried out at a flow rate of 2.0-2.5 l / ha. The last treatment is carried out on 05-10.09 15-20 days before harvesting. After harvesting in the early to mid-October, these areas are treated with the same biological product at a consumption rate of 3.0 l / ha.

The use of Trichoderma viride as a biological agent for the control of alternaria grapes in such a scheme, blocks to suppress current infection and reduce stock, provides stable and fairly high efficiency.

According to FB Hannibal et al. (Alternaria crops in Russia / F.B. Hannibal, A.S. Orina, M.M. Levitin.- Plant protection and quarantine, No. 5, 2010, p.30- 31) the disease caused by A. tenuissima is usually characterized by a high prevalence rate, but a low level of development and very rarely causes epiphytotics. A. tenuissima is a semi-parasite that is ubiquitous and infects mainly physiologically old or damaged tissues; under certain weather conditions, it causes mass infection, for example, of cabbage seed. The same researchers noted the harmfulness of alternariosis in various annual crops (potato, tomato, rape, sunflower, etc.), which is expressed in a decrease in the photosynthetic surface of leaves, in the moldiness of fruits and seeds, in a decrease in yield and contamination of agricultural products with mycotoxins and allergens.

More recently, the economically significant harmfulness of alternaria infections has been observed in perennial crops. So, according to Yakub G.V. (2003), the spread of alternaria apple tree - Alternaria alternata (Fr.:Fr.) Keissler, which in 2001 was noted as a new disease of this crop in the Western Ciscaucasia region (Evolutionary changes in phytopathogenic fungi populations in apple agrocenoses: scientific and applied aspects, is increasing / Math. Scientific and practical conference, SKZNIISiV. - Krasnodar: SKZNIISiV, 2003.- P.260-264). According to our data, in grapes (Anapo-Taman zone of the Krasnodar Territory), the development of alternariosis - Alternaria tenuissima (Kunze ex Pers.) Wiltshire, as a new harmful disease for this crop, has also been noted since 2000. Moreover, on hybrid varieties that occupy from 15 to 40% of vineyard area in various vineyards of the region, alternariosis has been developing epiphytotically for the last several years in a row since May in the phenophase “leafy leaf buds - two leaves opened”, ie affects young leaves. The problem is complicated by the complete absence of a scientifically based control system for this disease.

When developing such a system, special attention should be paid to the choice of protective equipment - a feature of anti-alternaria fungicides should be their lack of phytotoxicity to plants. Most chemical fungicides registered in the "List ..." against alternariosis in other crops are ineffective for alternariosis of grapes. In our studies, sufficient biological efficacy was observed only in analogues of strobilurins and folpet. Some chemical fungicides recommended for controlling alternariosis in other crops, in particular benzimidazoles, have a stimulating effect on the development of this disease, for example, in Mediterranean vineyards, as well as for alternaria cereals, cloves and strawberries (Mycosis of the vine / I.S. Popusoi, L.A. Margin. - Chisinau: Shtiintsa, 1989. - P.111). There is a report that the treatment of soybean with benomyl contributes to the development of diseases caused by A. alternata (General phytopathology / K.V. Popkova; 2nd ed., Revised and supplemented. - M.: “Drofa”, 2005. - P.72 )

When selecting strains from populations of A. tenuissima (Kunze ex Pers.) Wiltshire in various agroecological zones in various grape varieties of the Western Ciscaucasia region (Krasnodar Territory), 2 alternatives were identified as the most aggressive from strain 14 isolates, strain A 12/1 and A 02 / 3. In a pure culture, the antifungal activity of 12 biological preparations based on various types of bacteria and fungi (antagonists and hyperparasites), including several species of the genus Trichoderma, was tested on these two strains.

According to our studies, high antifungal activity against the causative agent of grape alternariosis, Alternaria tenuissima (Kunze ex Pers.) Wiltshire, was shown by the species T. viride, strain 4097, moderate activity - T. harzianum, strain 18 VIZR, weak activity - T. lignorum, strain 81-17 (Figure 1).

Based on the results of tests in a pure culture, field production experience was established and conducted in industrial vineyards of the Anapo-Taman agroecological zone of the Krasnodar Territory - the main one in the region's viticulture (AF Yuzhnaya LLC, Temryuk district). In an experiment on a Bianca grape highly susceptible to alternariosis, two biological products were tested: 1) based on T. viride, strain 4097; 2) based on T. harzianum, strain 18 VIZR.

Field tests have confirmed the relatively high biological effectiveness of the biological product based on the fungus Trichoderma viride strain 4097 in the suppression of A. tenuissima on grapes (table 2).

In the experiment, the biological effectiveness of the biological method for controlling grape alternariosis using biological products based on Trichoderma mushrooms — T. viride, strain 4097 and T. harzianum, strain 18 VIZR — was determined.

table 2 Biological effectiveness of biological methods of protecting grapes from alternariosis with the use of biological products based on various types of fungi p. Trichoderma and their impact on yield and product quality, Bianca variety, AF Yuzhnaya, 2010 Option Biological efficiency by dates of accounting,% Harvest from the bush, kg The total sugar content, g / 100 cm ³ 06/02/10 06/09/10 06/16/10 06/22/10 07/09/10 07/15/10 08/04/10 08/19/10 Standard 61.7 42.1 82.7 83.5 54.7 50.7 53,4 51.9 4.8 18.8 Trichodermin is new (T. viride, strain 4097) 70.9 78.0 75.6 89.6 80.3 77.1 76.7 72.1 6.5 20.5 Trichodermin (T. harzianum, strain 18 VIZR) 55.1 63,2 53.8 61.3 67.5 60,4 55,4 55.6 5,0 19,2 Control, R% 39.2 30,4 15.6 21,2 20.3 22.7 38.6 64.5 4.0 18,4 NDS ₀₅ - - - - - - - - 1.41 1.18

The standard option (Standard) was a chemical method of protection, which used synthetic organic preparations registered on grapes against other diseases and simultaneously registered on other cultures against alternariosis, because there are no registered fungicides in the "List ..." for use in Russia on grapes against alternariosis. In the control variant (Control), treatment against alternariosis was not carried out.

The use of a biological product based on the fungus Trichoderma viride, strain 4097, in the biological method of combating alternaria grapes can effectively and stably inhibit the development of this disease, without negatively affecting the grape plant itself. The biological effectiveness of the proposed method is 70.9-89.6%. Throughout the entire period of treatment with a biofungicide based on T. viride strain 4097, the effectiveness of the biological method of protection was higher than the standard treatment with chemical fungicides. Its effectiveness only after the use of strobilurin analogues was below the standard, but still quite high.

Claims (1)

The method of biological control of grape alternariosis, the causative agent of Alternaria tenuissima (Kunze ex Pers.) Wiltshire, characterized in that a biofungicide based on the fungus antagonist Trichoderma viride Pers. ex SF Gray strain 4097, titer of at least 1.0 · 10 ⁹ CFU / ml, consumption rate of 1.5-3.0 l / ha, the processing of which consists of 2 blocks: 1st - before harvesting to suppress current infection; 2nd after harvest to reduce the stock of infection.

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