RU2534561C2 - Method of treatment of potato tubers before placement for storage - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: method comprises treatment of potato tubers before placement for storage with biological product containing biomass Bacillus amyloliquefaciens RNCIM B-11008 with a titre of vegetative cells and spores of 1.24÷1.30×10¹⁰ CFU/ml and humates. At that the content of components of biological product is respectively 99 vol. % and 1.0 vol. %. The biological product is taken in a rate of application of 1.0 l/ton of potato tubers at a rate of application of working fluid of 10.0 l/ton.

EFFECT: inhibition of development of fungal and bacterial diseases of potato tubers during storage.

6 tbl, 5 ex

Description

The invention relates to agriculture, in particular to environmentally friendly methods of preventing the loss of stored potatoes from bacterial and fungal diseases, and can be used in technology for storing potatoes.

Product losses during post-harvest storage of fruits and vegetables are largely due to the development of bacterial and fungal diseases.

Known methods for the prophylactic treatment of potato tubers before laying for long-term storage with a rather limited number of chemical protective agents against phytopathogenic microflora based on carbendazim (Super Colfugo, Super Colfugo), fludioxonil (maxim) [see. Zeyruk V.N. Ways to reduce potato losses during storage / V.N. Zeyruk, K.A. Pshechenkov, I.I. Sidyakina // Plant protection and quarantine. - 2001. - No. 9. - S.40-41] and thiabendazole (fumigation fungicide whist) [see, Zeyruk V.N. Whist will protect potato tubers during storage / V.N. Shaldyaeva, V.M. Glez, K.A. Pshenchenkov, P.P. Galimov, L.M. Levit, V.A. F.Madyakin // Plant protection and quarantine. - 2006. - No. 2. - S. 61-62]. The restriction of the use of toxic and highly toxic chemical fungicides in the processing of stored food products is due to environmental requirements related to the ability of pesticides to accumulate in stored products in quantities exceeding sanitary standards.

There is a method of autumn dressing of tubers with a tank mixture, consisting of reduced doses of the preparations Maxim (50% of the dose) and extrasol (10 L of a 5% solution / t of tubers) [see, Petrov VB The use of microbiological preparations - an indispensable element of intensive technologies in potato growing / V. B. Petrov, V. K. Chebotar // Potatoes and vegetables. - 2011.-№8. - S.18-21]. The need to make tank mixtures significantly complicates the process of etching in production conditions and requires the observance of additional safety measures when working with the chemical component of the mixture.

Modern environmentally friendly technologies for storing crop products involve the use of environmentally friendly bacterial preparations and their metabolic products.

Known biological product to stimulate the growth and protection of plants from diseases, increase productivity and soil fertility based on Bacillus amyloliquefaciens VKPM B-11008 [RF Patent No. 2478290, IPC A01N 63/02, A01C 1/06, C12N 1/20, C12R 1/07 publ. 04/10/2013]. The biological product contains Bacillus amyloliquefaciens VKPM B-11008 biomass (vegetative cell titer and spores 1.24 ÷ 1.30 × 10 ¹⁰ CFU / ml, spore content 94% of the total CFU (99 vol.%) And humates (1.0 vol. .%). The process of obtaining a biological product consists in the deep cultivation of the producer strain B. amyloliquefaciens VKPM B-11008 on a nutrient medium with wheat bran (composition, g / l: wheat bran - 40.0; corn extract - 0.5; MgSO ₄ - 0.9; KH ₂ PO ₄ -0.5; CaCO ₃ - 1.0; pH 7.0 ± 0.2) followed by the addition of humates.Information on the method of using the biological product for processing potato tubers before storage The knowledge and norms of consumption of biological product and working fluid during potato processing are not available in available sources of information.

The closest invention to the achieved result, taken as a prototype, is a method of processing potato tubers before storing the biological product phytosporin-M in liquid form. In accordance with the data specified in the list of pesticides and agrochemicals approved for use in the Russian Federation, the rate of consumption of a biological product is 100 ml / l of water with a flow rate of 1.0 l / 10 kg of tubers [see, List of pesticides and agrochemicals approved for use on the territory of the Russian Federation. 2012: Reference publication, an appendix to the journal "Protection and Plant Quarantine" No. 4, 2012. - M., 2012. - P.134], which in terms of 1 ton of potato tubers is 10 l of the drug with a working fluid flow rate of 100 l This is one of the most significant disadvantages of the prototype, since a high dose of the drug when processing potato tubers before storage increases the level of additional costs, reducing profitability, and processing vegetable products with a significant volume of working solution (100 l / t) leads to the need for long drying of tubers in production conditions. Another important disadvantage of the prototype is the narrow spectrum of diseases suppressed during storage of tubers, which, in accordance with the manufacturer’s registration data, is limited to rhizoctonia, dry fusarium rot, wet bacterial rot and phomosis [see, List of pesticides and agrochemicals approved for use in the Russian Federation . 2012: Reference publication, an appendix to the journal "Protection and Quarantine of Plants" No. 4, 2012. - M., 2012. - P.134].

The objective of the invention is the development of an environmentally friendly method of processing potato tubers before bookmarking for protection against fungal and bacterial diseases with a high level of biological, economic and economic efficiency and low consumption rates.

The technical result of the invention is a partial or complete inhibition of the development of diseases of potato tubers during storage, increasing the profitability of the storage process due to low consumption rates of the biological product and a high level of potato tuber preservation.

The technical result in the claimed invention is achieved by the fact that in the method of treating potato tubers before storage, involving surface treatment with a biological product according to the invention, a biological product is used containing Bacillus amyloliquefaciens VKPM B-11008 biomass with a titer of vegetative cells and spores 1.24 ÷ 1.30 × 10 ¹⁰ CFU / ml (99 vol.%) And humates (1.0 vol.%), At a rate of consumption of 1.0 l / t of potato tubers at a rate of flow of working fluid of 10.0 l / t.

The method is implemented as follows.

A biological product based on Bacillus amyloliquefaciens VKPM B-11008, containing 1.24 ÷ 1.30 × 10 ¹⁰ CFU / ml of vegetative cells and spores (99 vol.%) And humates (1.0 vol.%), In a volume of 1 liter per 1 ton of tubers is mixed with 9 l of tap water, receiving a volume of working fluid of 10 l / t of tubers. The area of the treated tuberous surface is at a rate of consumption of a biological product of 1 l / t and a working fluid of 10 l / t, corresponding to the claims, at least 90%. The processing of tuberous material is carried out using standard agricultural equipment intended for the treatment of tubers (PMK, PUM-30, etc.). Before storing potatoes in the vegetable store, the tubers, if necessary, are briefly dried naturally. Storage of treated tubers is carried out in bulk (in bulk) in a stationary vegetable storehouse of a sectionless type or in storage of a sectional type. During storage, the embankment maintains a constant temperature (+ 2 ... + 4 ° C) and humidity (85-90%).

The undoubted advantage of the proposed method for processing potato tubers before laying them for storage are low consumption rates of the biological product (1.0 l / t of tubers) and working fluid (10 l / t of tubers), in contrast to the prototype (100 ml of biological product / l of water at a consumption rate working fluid 1.0 l / 10 kg tubers). This leads to a significant reduction in the level of additional costs for the purchase of a biological product, simplification of the production dressing procedure for tubers and a reduction in the drying period after processing, which ultimately, in accordance with the claimed invention, increases the cost-effectiveness of the protective measure. With a decrease in the claimed invention, the norms of consumption of a biological product of less than 1 l / t of tubers and working fluid of less than 10 l / t, the technical result is not achieved.

As an advantageous characteristic of the claimed invention, the control of both surface contamination of potato tubers by phytopathogenic micromycetes and the number of phytopathogens inside stored potato tubers is considered. At the same time, the effectiveness of the action against external phytopathogenic microflora is comparable to the action against infection inside plant tissues. This is due to the necessary and sufficient number of viable cells for effective and uniform colonization of the tuberous surface, which when processed with a biological product in accordance with the claimed invention, at a rate of consumption of a biological product of 1 l / t of tubers and a working fluid of 10 l / t is at least 90%.

It should be noted that the processing of potato tubers before storage in accordance with the claimed invention leads to a significant expansion of the spectrum of suppressed diseases, which includes an almost complete list of the most harmful diseases of stored potatoes and their pathogens:

late blight (Phytophthora infestans); common scab (Streptomyces scabies), silver (Helmintosporium atrovirens), tuberous (Oospora pustulans) and black (Rhizoctonia solani); dry rot (fungi of the river Fusarium) and rubber (Geotrichum candidum); phomosis (Phoma exiqua); bacterial ring rot (Clavibacter muchilaginosus subsp.sepedonicum), as well as molds on the eyes of various etiologies (fungi of the genera Penicillium, Aspergillus, Mucor, Aureobasidium, Chaetomium).

The inventive method of processing potatoes before storage does not include the use of chemical pesticides, which allows agricultural producers to sell environmentally friendly products that are in high demand.

The achievement of the technical result of the invention is demonstrated by the following examples.

Example 1, demonstrating the high biological efficiency of processing potato tubers of the claimed method in comparison with the prototype

 The processing efficiency of potato tubers in accordance with the claims is evaluated on the medium early variety Nevsky. In the experiments, tubers are used that meet the requirements and standards of GOST 7176-85 [see GOST 7176-85 Fresh, food, harvested and delivered potatoes / Publishing house of standards: M., 2004. - 6 pp.].

When determining diseases, the main signs of diseases of potato tubers are taken into account [see, Khokhryakov, M.K. Determinant of plant diseases / K.M. Khokhryakov, T.L. Dobrozrakova, K.M. Stepanov, M.F. Letova. - St. Petersburg: Doe, 2003. - 592 pp.] And calculate the content of tubers with each type of disease as a percentage of the total sample analyzed [see GOST 7194-81 Fresh potatoes. Acceptance rules and methods for determining quality / Publishing house of standards, M. - 36 p.]. To detect a latent infection, the surface of apparently healthy tubers is washed, sterilized with a swab moistened with alcohol, sterile instruments are cut out surface pieces of tissue and eyes with the appropriate tissue and placed in Petri dishes on acidified lactic acid to pH 5.5 Chapek's medium [see, Semenov S. M. Laboratory media for actinomycetes and fungi: Reference book / S.M.Semenov. - M .: Agropromizdat, 1990. - p.172] and potato-glucose medium [see, Semenov, S.M. Laboratory media for actinomycetes and fungi: Reference book / S.M.Semenov. - M .: Agropromizdat, 1990. - p. 192]. After 7-10 days, sporulating mycelia of phytopathogenic fungi are isolated into a pure culture and identified in accordance with the determinants [see, Khokhryakov M.K. Determinant of plant diseases / K.M. Khokhryakov, T.L. Dobrozrakova, K.M. Stepanov, M.F. Letova. - SPb .: Doe, 2003 .-- 592 p .; see, Sutton, D. Identifier of pathogenic and conditionally pathogenic fungi / D. Sutton, A. Fotergill, M. Rinaldi. - M .: Mir, 2001. - 468 p.]. According to the criterion of reducing the prevalence of diseases, the biological effectiveness of treating tubers by the claimed method (at a dose of 1.0 l / t and a flow rate of 10 l / t) is evaluated in comparison with the prototype (at a dose of 100 ml / l of water and a flow rate of 1.0 l / 10 kg of tubers) [see List of pesticides and agrochemicals approved for use in the Russian Federation. 2012: Reference publication, an appendix to the journal "Protection and Quarantine of Plants" No. 4, 2012. - M., 2012. - P.134]. In the control embodiment, the tubers are treated with water. The mass of tubers in each version is 6.6 tons. The storage of tubers is carried out in a stationary vegetable storage section type for 180 days. During storage, maintain a constant temperature (+ 2 ... + 4 ° C) and humidity (85-90%).

The treatment of tubers by the claimed method reduces the prevalence rate of diseases of potato tubers when stored in a vegetable store according to the sectional type for 180 days by 1.8-6.5 times compared with the control depending on the type of pathogen and completely inhibits the development of silver scab (Helmintosporium atrovirens) rubber rot (Geotrichum candidum) and phomosis (Phoma exiqua) (table 1). The effectiveness of reducing the prevalence of diseases in the variant with the prototype is significantly lower and does not exceed an average of 1.3-2.7 times depending on the disease compared with the control.

Table 1 The effect of the processing of potato tubers of Nevsky variety by the claimed method in comparison with the prototype on the prevalence rate of potato diseases during storage for 90 (in the numerator) and 180 (in the denominator) days Disease and pathogen Prevalence,% Initial level before storage The control The processing of the claimed method Prototype processing Blight (Phytophthora infestans) 11.5 16.8 / 21.5 12.0 / 12.2 15.7 / 16.5 Scab (Streptomyces scabies) 25.0 49.0 / 64.6 26.5 / 28.3 31.2 / 38.0 Silver scab (Helmintosporium atrovirens) 1,6 4.0 / 4.9 but / but 2.7 / 3.1 Tuberous scab (Oospora pustulans) 5.8 15.2 / 37.5 6.0 / 6.6 16.0 / 18.4 Black scab (Rhizoctonia solani) 15.0 31.0 / 53.8 15,5 / 16,2 25.5 / 26.9 Dry rot of tubers (mushrooms of the river Fusarium) 10.0 28.5 / 69.7 10.0 / 10.7 30.2 / 34.8 Rubber rot (Geotrichum candidum) 1,6 2.9 / 4.1 but / but 2.4 / 3.0 Phomosis (Phoma exiqua) 3,5 4.3 / 7.0 but / but 4.0 / 4.0 Molds on the eyes (fungi of the genera Penicillium, Aspergillus, Mucor, Aureobasidium, Chaetomium) 26.6 57.0 / 95.2 26.8 / 27.7 33.7 / 35.4 Ring rot (Clavibacter muchilaginosus subsp.sepedonicum) 3.3 16.2 / 21.8 4.1 / 4.4 8.0 / 10.9 Note: but - not detected.

According to the results of accounting for absolutely rotten tubers selected in the storage process and the proportion of diseased tubers in the stored potato mass, the treatment of tubers with the claimed method reduces the overall losses from diseases when stored in a vegetable storehouse in bulk in a sectional type for 180 days by 3.5 times compared with the control , ensuring the preservation of tubers at the level of 78.4% against 24% in the control, and 1.9 times compared with the processing of tubers by the prototype, exceeding the level of tuber preservation by 36.9% (table 2).

table 2 The safety of potato tubers processed by the claimed method compared with the processing of the prototype Experience option Shelf life, days The proportion of rotted and diseased tubers,% The share of healthy tubers,% The control 90 57.9 42.1 Prototype processing 42,4 57.6 The processing of the claimed method 19.5 80.5 The control 180 76.0 24.0 Prototype processing 58.5 41.5 The processing of the claimed method 21.6 78,4

Example 2, demonstrating the effect of the proposed method for processing potato tubers before storage on the starch content

The processing of the inventive method and the prototype is carried out in accordance with example 1. The starch content in the tubers is determined by the photoelectrocolorimetric method, the essence of which is the hydrolysis of starch with diluted sulfuric acid and photoelectrocolorimetric determination of the color intensity of the solution of the bluish-green compound of anthrone with glucose, followed by quantitative conversion to starch [see ., GOST 7194-81 Fresh potatoes. Acceptance rules and methods for determining quality / Publishing house of standards, M. - S.24-30]. The treatment of tubers before storage by the claimed method, in contrast to the processing of the prototype, helps to maintain the initial level of starchiness of tubers and increase the starch content by 180 days of storage by 29.4% compared with the control variant and by 9.1% compared with the processing of the prototype (table 3).

Table 3 The starch content in potato tubers during storage of tubers processed by the claimed method in comparison with the processing of the prototype Experience option The starch content,% 90 days 180 days The control 12.1 10,2 Prototype processing 13.0 12.1 The processing of the claimed method 13,2 13,2

Example 3, demonstrating the influence of the proposed method for processing potato tubers on the number of phytopathogenic micromycetes on the surface of tubers

The treatment of tubers is carried out in accordance with example 1. The number of micromycetes that populate the surface tissues of tubers is counted by direct microscopy and plating of multiple dilutions of the suspensions obtained by washing from the surface of the tubers on nutrient media: Chapek's medium [see, S. Semenov, M. Laboratory media for actinomycetes and fungi: Reference book / S.M.Semenov. - M .: Agropromizdat, 1990. - p.172] and potato-glucose medium [see, S. Semenov. Laboratory media for actinomycetes and fungi: Reference book / S.M.Semenov. - M .: Agropromizdat, 1990. - P.192].

As can be seen from the data presented in table 4, the processing of the claimed method of potato tubers prevents the development of pathogenic micromycetes on the tuberous surface, which is expressed in a decrease in their number by the end of the storage period (180th day) by 15.7 times compared with the control and 3.7 times compared with the processing of the prototype at the initial level of surface contamination of tubers with phytopathogenic fungi before laying for storage - 0.17 × 10 ⁵ CFU / g.

Table 4 The number of micromycetes on the surface of Nevsky potato tubers during a storage period of 180 days Option The number of micromycetes, × 10 ⁵ CFU / g 30 days 60 days 90 days 180 days The control 0.5 1.35 3.6 26.7 The processing of the claimed method 0.3 0.3 0.8 1.7 Prototype processing 0.35 0.8 2,57 6.4

Example 4, demonstrating the influence of the proposed method for the treatment of potato tubers on the number of phytopathogenic micromycetes in the internal tissues of tubers

The treatment of tubers is carried out in accordance with example 1. To account for the number of micromycetes that inhabit the internal tissues of the tubers, an average weighed sample of 5 g of tuber material crushed in a porcelain cup is prepared to a puree state, diluted with water and a multiple dilution of the suspension is inoculated onto nutrient media, followed by identification and microbiological calculation of the number of grown colonies of micromycetes in terms of the coefficient of dilution of the suspension [see, Methods of soil microbiology and biochemistry: Textbook / Ed. D.G. Zvyagintsev. - M.: Publishing House of Moscow State University, 1991. - 310 p.].

As can be seen from the data of table 5, the processing of potato tubers before storage by the claimed method leads to a decrease in the number of micromycetes by the end of the storage period (180th day) 13.4 times compared with the control and 5.3 times compared with the processing of the prototype.

Table 5 The number of micromycetes in the internal tissues of tubers of the Nevsky potato variety during the storage period of 180 days Option The number of micromycetes, × 10 CFU / g 30 days 60 days 90 days 180 days The control 0.26 0.81 2,3 18.3 The processing of the claimed method 0.09 0.17 0.79 1.37 Prototype processing 0.15 0.34 1.12 7.3 Note: the initial level of internal infection of tubers with phytopathogenic fungi before laying for storage is 0.08 × 10 ³ CFU / g

Thus, based on the quantitative characteristics of the number of micromycetes on the outer surface and in the internal tissues, it can be argued that the decrease in surface contamination under the influence of a biological product compared with the control during storage does not occur due to the penetration of phytopathogens into the tissues and increased internal infection, but by complete or partial suppression of exo- and endopathogenic fungal microflora.

Example 5, indicating the high economic efficiency of the proposed method for processing tubers when laying for storage in comparison with the processing of the prototype

The treatment of tubers is carried out in accordance with example 1. Economic efficiency is calculated as the cost difference of the stored tuber material between the options, taking into account the average selling price of 1 ton of potato according to 2013 data (13 thousand rubles / ton) and the potato mass in each variant is 6 , 6 t.

The processing of potato tubers by the claimed method of potatoes in the norm of 1.0 l / t, followed by storage in a vegetable storehouse in bulk according to the section type for 180 days, provides a conditionally net income of 6.87 thousand rubles / t, which exceeds the economic efficiency of processing tuber prototype 3.29 times (table 6).

Table 6 The economic efficiency of processing potato tubers of the Nevsky variety of the claimed method compared to the processing of the prototype Option The safety of potatoes,% Additional safety of potatoes, t / thousand rubles. Conditional net income, RUB / t Payback of additional costs, times The control 24.0 - - - Prototype processing 41.5 1.16 / 15.08 2085 10,4 The processing of the claimed method 78,4 3,59 / 46,67 6870 34,4

Thus, the inventive method of processing potato tubers before laying them for storage provides a significant reduction or complete inhibition of the development of diseases of potato tubers during storage, increasing the profitability of the storage process due to the low consumption rates of the biological product and the high level of preservation of potato tubers.

Claims (1)

A method of processing potato tubers before storing, which consists in surface treatment of potato tubers with a biological product, characterized in that a biological product containing Bacillus amyloliquefaciens VKPM B-11008 biomass with a titer of vegetative cells and spores of 1.24 ÷ 1.30 × 10 ^{10 is} used for processing CFU / ml (99 vol.%) And humates (1.0 vol.%), Taken at a rate of consumption of 1.0 l / t of potato tubers at a rate of flow of working fluid of 10.0 l / t.