RU2767346C1 - Method for pre-planting treatment of potato tubers - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention is a method for pre-planting treatment of potato tubers, includes treatment of tubers with a microbial culture, wherein for pre-planting treatment of tubers used as microbial culture Pseudomonas extremorientalis with titre 0.5×10⁷–1.5×10⁸ cells/ml with addition of bentonite into said microbial culture in amount of 0.1–3 wt%, wherein treatment of tubers is carried out at rate of 100 ml of the obtained preparation per 10 kg of potatoes.

EFFECT: invention allows increasing potato yield.

3 cl, 5 tbl, 1 ex

Description

The invention relates to agriculture and can be used in the cultivation of potatoes. EFFECT: invention makes it possible to increase the yield of potatoes due to the growth-stimulating and fungistatic effects exhibited by bacteria.

Potato is one of the most important vegetable crops in the domestic crop industry. The use of microbial preparations capable of inducing non-specific potato resistance reactions to many diseases of fungal, bacterial and viral origin is one of the most urgent and timely tasks of modern domestic biotechnology, as well as a promising resource for increasing potato yields. The use of regional resources of clay minerals also makes it possible to solve the problems of lack of mineral fertilizers and trace elements in soils.

A known method of presowing treatment of seeds from the patent of the Russian Federation No. 2243639, A01C 1/00, A01C 1/06, publ. February 20, 2005 [1], which provides for the sequential application of an aqueous suspension of an adhesive substance, finely ground peat in an amount of 0.3-0.5 of the mass fraction of seeds to the seeds, the introduction of macro-, micro-fertilizers and a growth stimulator into the aqueous suspension of the adhesive substance, application to the coated peat seeds by alternating a mixture of bentonite with peat in the ratio of their mass fractions of 0.14-0.64 and an aqueous suspension of adhesive with macro-microfertilizers and a growth stimulator, followed by running them in a pan and drying.

The disadvantage of this method is a complex technological chain of sequential application of various reagents to the seeds, as well as the use of an adhesive when applying them to the seeds.

Known biological product Azogran, which is a component preparation of the clay mineral bentonite, nitrogen-fixing strain Azotobacter vinelandii IMV B-7076 and phosphate-mobilizing bacteria Bacillus subtilis IMV B-7023 on barley from the article: Skorokhod I.A., Ulziyzhargal E., Kurdish I.K. ., Gorgo Yu.P. The effect of the nanocomposite biopreparation azogran on barley seeds subjected to oxidative stress // Scientific Light (Wroclaw, Poland) No. 36, 2020. P. 10-14. [2]. The number of viable cells in the biological product was: A. vinelandii IMV B-7076 - (3.1±0.1)×10 ⁷ cells/ml; B. subtilis IMV B-7023 - (3.4±0.1)×10 ⁷ cells/ml. According to the results of experiments in the greenhouse, it was found that post-treatment with a nanocomposite complex bacterial preparation of hydrogen peroxide-stressed seeds of spring barley cv. Virage significantly stimulated the growth of plants at different stages of development. It has been shown that post-stress treatment of seeding material of barley cv. Virazh with nanocomposite complex bacterial biological preparation Azogran was accompanied by an increase in the number of grains per ear by 22.8%, in comparison with this indicator for plants grown from stressed seeds.

The disadvantage of this method is the use of the drug Azogran for seeds stressed with hydrogen peroxide. It is not known whether the drug affects normally developing plants and whether it increases the number of grains per spike.

A known method of pre-sowing treatment of seeds of spring cereals (options) from the patent of the Russian Federation No. 2655789, A01N 63/02, publ. May 29, 2018 [3].

A method for presowing seed treatment of spring cereals, including seed treatment with at least one microbial culture. As a microbial culture, the following are used: Pseudomonas sp., Bacillus sp., phosphate-mobilizing bacteria isolated from coprolites of earthworms Eisenia fetida. At the same time, seed treatment is carried out with a solution of microbial culture with a concentration of 10 ⁷ -10 ⁸ cells/ml at the rate of 100 w.h. drug per 10,000 v.h. seeds. This method of pre-sowing treatment of seeds of spring cereals helps to reduce the infestation of spring crops with pathogens of root rot, with a subsequent increase in yield and improvement in grain quality.

The disadvantage of this method is the use of two microbial cultures simultaneously in the preparation, which makes it technologically complex.

Closest to the claimed invention is a method of preplant treatment of potato tubers, known from the patent of the Russian Federation No. 2663335, A01C 1/00, publ. 08/03/2018 [4].

In the method of preplant treatment of potato tubers, including preplant treatment of tubers with bacteria with fungistatic and growth-promoting properties, an enrichment culture of Pseudomonas sp strain is used. with a concentration of 10 ⁶ -10 ⁷ cells / ml of the working suspension, used at the rate of 30 l / t of tubers. For processing, a strain of Pseudomonas sp. isolated from coprolites of earthworms Eisenia fetida is used. Potato tubers after treatment with the drug are dried for 2-3 hours. EFFECT: invention allows to increase the yield of potatoes by reducing the prevalence and development of potato diseases.

The disadvantage of this method is a large amount of the drug for the treatment of potato tubers (30 l/t of tubers), as well as insufficiently proven viability of bacteria on tubers and their development in the potato rhizosphere.

The technical objective of the present invention is to develop a method for pre-planting treatment of potato tubers to protect against certain phytopathogens and increase the marketability and yield of potatoes due to the growth-stimulating and fungistatic effects manifested by bacteria, which improve the morphometric parameters of plants and reduce the infestation of potatoes by phytopathogens during the growing season, as well as improving the safety of bacteria on surface of tubers by adding bentonite to the composition of the preparation for treating tubers.

The specified technical result is achieved by the fact that in the method of preplant treatment of potato tubers, including the treatment of tubers with a microbial culture, while for preplant treatment of tubers, Pseudomonas extremorientalis is used as a microbial culture with a titer of 0.5×10 ⁷ -1.5×10 ⁸ cells/ ml with the addition of bentonite to the specified microbial culture in the amount of 0.1-3 wt.%, while the treatment of tubers is carried out at the rate of 100 ml of the resulting preparation per 10 kg of potatoes.

For the preparation of the preparation, a strain of Pseudomonas extremorientalis isolated from coprolites of earthworms Eisenia fetida and bentonite of the PBMB brand are used.

The essence of the invention is as follows.

The bacterium Pseudomonas extremorientalis used in the composition of the biological product belongs to the group of rhizobacteria, which are natural symbionts of agricultural plants. The use of biopreparations based on them induces reactions of nonspecific plant resistance to many diseases. In addition, the bacteria Pseudomonas extremorientalis produce substances of a growth-stimulating nature that improve the morphometric parameters of plants and thereby provide an increase in yield and product quality. To improve the safety of bacteria on the surface of tubers, bentonite is added to the preparation for treating tubers.

Pure cultures of the studied microorganisms Pseudomonas extremorientalis were obtained by isolating earthworms Eisenia fetida from coprolites, for which direct inoculation was carried out from a mixed average sample of coprolite on meat-peptone agar, followed by a series of passages on the same medium and final re-seeding into test tubes on oblique agar.

Accumulative cultures of bacteria were obtained by submerged periodic cultivation of bacteria isolated in a pure culture in King B medium in flat-bottomed flasks with a volume of 250 ml with 100 ml of medium in a thermostatically controlled shaker. Cultivation temperature +28…30°С, duration - 3 days. Aeration was carried out by stirring the medium on a rocking chair (180-200 rpm). Cultivation was continued until the bacterial titer reached 1×10 ⁹ cells/ml. The abundance control was carried out by standard methods: direct counting in the Goryaev chamber and counting according to the Koch method.

An effective carrier for P. extremorientalis bacterial cells is bentonite, which, if added to a microbial preparation, provides a higher cell titer during the storage period and, at the same time, is characterized by the best adsorption on the surface of potato tubers.

Used bentonite brand PBMB (LLC "Bentonite Khakassia") [www.b-kh.ru], which is a fine powder of light gray color. Mineral composition: montmorillonite, kaolinite, hydromica, quartz, alkali feldspar, mica, calcite. The average chemical composition of bentonite in%: SiO ₂ - 59.68; Al ₂ O ₃ - 18.63; Fe ₂ O ₃ - 3.93; CaO - 2.769; MgO - 2.43; K ₂ O - 1.62; Na ₂ O - 0.98; FeO - 0.67; TiO ₂ - 0.59; SO ₃ - 0.16; P ₂ O ₅ - 0.12; MNO - 0.05. The presence in the composition of bentonite of the main nutrients (potassium, phosphorus), macroelements (calcium, magnesium) and microelements (manganese) in a water-soluble form contribute to the stimulation of the growth processes of potato plants.

Example 1

It is necessary to take 0.5 ml of culture with a titer of 1 × 10^ninecells/mlPseudomonas extremorientalis,add 99.5 ml of distilled water and 0.1 g bentonite.

The remaining examples of the preparation of the preparation for the treatment of potato tubers are obtained in accordance with table 1.

Table 1 Examples of the preparation of a bacterial preparation with bentonite for the treatment of potato tubers

No. Examples The volume of culture with a titer of bacteria 1.0×10 ⁹ cells/ml, ml The volume of distillate
lyirovan-
noah water, ml The resulting titer of bacteria in the preparation cells / ml Amount of bentonite, g one Bacteria with bentonite 0.5 99.5 0.5×10 ⁷ 0.1 2 Bacteria with bentonite 1.0 99.0 1.0×10 ⁷ 0.2 3 Bacteria with bentonite 2.5 97.5 0.25×10 ⁸ 0.5 4 Bacteria with bentonite 7.5 92.5 0.75×10 ⁸ 1.5 five Bacteria with bentonite 10.0 90.0 1.0×10 ⁸ 2.0 6 Bacteria with bentonite 15.0 85.0 1.5×10 ⁸ 3.0 7 Control (comparison option) 0 100.0 0 0 8 Bacteria without bentonite (prototype) 10.0 90.0 1.0×10 ⁸ 0

Examples

1. Pre-planting treatment of potato tubers with microbial culture P. extremorientalis on a solid carrier (0.1 wt.% bentonite), bacterial titer - 0.5×10 ⁷ cells. /ml;

2. Pre-planting treatment of potato tubers with microbial culture P. extremorientalis on a solid carrier (0.2 wt.% bentonite), bacterial titer - 1×10 ⁷ cells. /ml;

3. Pre-planting treatment of potato tubers with microbial culture P. extremorientalis on a solid carrier in diluted form (0.5 wt.% bentonite), bacterial titer - 0.25×10 ⁸ cells/ml.

4. Pre-planting treatment of potato tubers with microbial culture P. extremorientalis on a solid carrier in a diluted form (1.5 wt.% bentonite), bacterial titer - 0.75×10 ⁸ cells/ml.

5. Pre-planting treatment of potato tubers with microbial culture P. extremorientalis on a solid carrier in a diluted form (2 wt.% bentonite), bacterial titer - 1×10 ⁸ cells/ml.

6. Pre-planting treatment of potato tubers with microbial culture P. extremorientalis on a solid carrier in diluted form (3 wt.% bentonite), bacterial titer - 1.5×10 ⁸ cells/ml.

7. Control without treatment of potato tubers with the drug (spraying with distilled water) (comparison variant)

8. Pre-planting treatment of potato tubers with P. extremorientalis microbial culture, bacterial titer - 1×10 ⁸ cells. /ml (prototype processing)

Before planting, potato tubers of the Pamyati Rogacheva variety were sprayed using a knapsack sprayer with a working solution of the drug, the active ingredient of which is the bacteria Pseudomonas extremorientalis and bentonite at the rate of 100 ml of the resulting drug per 10 kg of potato tubers.

Further, the treated potato tubers were dried in a shaded place (not in direct sunlight) for 2-3 hours. Landing pattern 70×40cm. Planting density - 35.7 thousand tubers per hectare. In the control variant, the tubers were treated with distilled water.

The experiment was carried out on gray forest medium-podzolized soil. Soil tillage for planting potatoes included autumn plowing and spring cultivation. Harvesting of plots and accounting of the yield were carried out at the onset of the physiological maturity of the potato (withering away of the green mass).

The effectiveness of the use of a bacterial preparation based on Pseudomonas extremorientalis was evaluated in September in terms of yield and tuber analysis by fractions. After a month of storage, a tuber phytoanalysis was performed.

According to the results of the field experiment, late blight and rhizoctoniosis showed itself to the greatest extent among fungal pathogens in the phase of active potato vegetation. An analysis of the prevalence and degree of damage by late blight in the phase of potato budding showed that presowing bacterization of P. extremorientalis tubers with bentonite in all cases reduced the prevalence and degree of damage by late blight. The same trend is observed in relation to Rhizoctonia (Table 2).

Table 2 - Phytopathological assessment of the state of potato plants in the budding phase - the beginning of flowering

Example Prevalence, % Degree of damage, % late blight one 84.0 19.6 2 95.0 13.4 3 81.2 15.0 4 80.0 16.4 five 82.5 18.8 6 82.3 18.6 7 85.0 34.4 8 87.5 24.6 Rhizoctonia one 5.0 2 5.0 - 3 4.5 - 4 4.5 - five 5.0 - 6 4.5 - 7 7.5 - 8 2.5 - Note . "-" - the calculation of such indicators for this disease is not carried out

Under the influence of bacteria and bentonite applied to tubers, the number of tubers on one plant increases by 1.3-2.6 pieces, the total mass of tubers of one plant by 64.7-127.9 g (table 3). Due to this, the biological yield increases by 14-30%.

Table 3 - The effect of bacterization and the introduction of bentonite on the yield and structure of the potato crop

Example Total number of tubers, pieces / 1 plant The total mass of tubers on the 1st plant, g Average weight of the 1st tuber, g Biological yield, t/ha one 10.85 519.49 47.88 19.94 2 10.97 526.17 47.96 20.19 3 11.00 534.60 48.60 20.52 4 11.01 535.97 48.68 20.57 five 9.71 472.80 48.69 18.14 6 9.68 468.03 48.35 17.96 7 8.38 408.12 48.70 15.66 8 10.57 522.14 49.40 20.04

Fractional analysis of the potato crop showed that all examples of bacterization with bentonite contribute not only to an increase in the yield of potatoes, but also to an increase in the fractions of seed, food and commercial potatoes in its composition and, as a result, an increase in the marketability of potatoes. Gross yield increased by 15-31%. The yield of seed potatoes due to bacterization increased by 19-43%. The yield of ware potatoes under the influence of the studied methods of treating tubers with bacteria and bentonite increased by 15-32%. The yield of marketable potatoes in the experimental variants increased by 16-33% (table 4).

Table 4 - The influence of the studied methods of bacterization on the yield of potatoes by fractions

Example Harvest Marketability Gross seed potatoes Food
(from 35 to 80 mm across) Commodity
(gross minus small) t/ha t/ha t/ha t/ha % one 19.94 18.98 18.54 19.54 98.0 2 20.19 19.45 18.76 19.80 98.1 3 20.52 19.72 18.98 20.17 98.3 4 20.57 19.00 19.05 20.22 98.3 five 18.14 16.16 16.79 17.79 98.1 6 17.96 15.81 16.59 17.62 98.1 7 15.66 13.24 14.34 15.18 96.9 8 20.04 18.89 18.52 19.68 98.2

Tuber analysis, carried out a month after harvest, showed that at this stage the potato is characterized by a low level of infection by such pathogens as black leg, ring rot and late blight. At the same time, all the studied methods of processing tubers ensured a decrease in the percentage of tubers affected by the most dangerous diseases, such as late blight, ring rot and dry rot (table 5).

Table 5 - Results of phytosanitary analysis of potato tubers after a month of storage

Example Total, % Infection of potato tubers, % Late blight, % Black leg, % Ring-
rot, % Dry rot, % Rizok-
toniosis Scab usually
venous Scab
spiny,
% one 41 0 0 0 2 13 13 13 2 41 0 one 0 3 eleven fourteen 12 3 40 0 0 0 2 12 fourteen 12 4 42 0 0 0 4 13 fourteen eleven five 49 2 0 0 4 21 eleven eleven 6 47 one one 0 4 twenty 10 eleven 7 44 3 0 one five 19 6 10 8 63 0 2 0 0 26 22 13

Thus, the pre-planting treatment of potato tubers with the Pseudomonas extremorientalis microbial culture with the addition of bentonite in the field experiment contributed to a 15-31% increase in yield, an increase in the share of marketable potatoes and a decrease in tuber damage by pathogens.

Claims (3)

1. A method for preplant treatment of potato tubers, including treatment of tubers with a microbial culture, characterized in that for preplant treatment of tubers, Pseudomonas extremorientalis with a titer of 1.0-1.5×10 ⁸ cells/ml is used as a microbial culture with the addition of bentonite to the specified culture in an amount of from 0.1 to 2.0 wt.%, while the treatment of tubers is carried out at the rate of 100 ml of the resulting preparation per 10 kg of potatoes. 2. The method according to claim 1, characterized in that a strain of Pseudomonas extremorientalis isolated from coprolites of earthworms Eisenia fetida is used. 3. The method according to claim 1, characterized in that PBMB grade bentonite is used.