RU2028054C1 - Method for manufacture of means of combatting herbivorous insects - Google Patents

Abstract

FIELD: agriculture. SUBSTANCE: this method prescribes cultivating yellow-green alga called Nephrochloris salina C. on nutritive medium and additionally introducing microalga Platymonas viridis R. for effectively combatting herbivorous insects, particularly, Colorado potato beetle and white American butterfly. EFFECT: novel technique.

Description

 The invention relates to agriculture and is intended to combat herbivorous insects - the Colorado potato beetle and the white American butterfly at the younger stage of the larval phases.

 It is known to use an autolysate of a two-month-old culture of the alga Platymonas viridis Rouch (see ed. St. N 1095483, class A 01 N 63/00), formed during the aging of the culture, as an inhibitor of the nutrition of lepidopteran insects.

 It is also known to use an autolysate of the alga Nephrochloris salina Carter as an entomocidal agent obtained in a similar manner. It is noted that the maximum concentration of cells in the culture is observed on the 25-30th day, and complete lysis on the 45-60th day.

 The disadvantages of the known methods include low adaptability; the mismatch of the period of high biological activity of autolysates with a peak of harmfulness of herbivorous insects and optimal processing times of plants, as well as the difficulty of application in production conditions associated with difficulties in transportation and storage.

 The aim of the invention is to increase the manufacturability and effectiveness of the funds and the cost-effectiveness of its application by using intensive technology of continuous cultivation.

 The goal is achieved by using a suspension of a continuous mixed culture of P.viridis and N.salina for processing plants.

 Comparative analysis with the prototype shows that the practical use of intensive continuous mixed culture of N.salina and P.viridis is unknown, so the claimed method meets the criteria of the invention of "novelty" and "significant differences".

The method is implemented as follows. Cultivation is carried out in flat cultivators with a capacity of 1.0 (depending on the task, the volume can be increased). Typical cultures from the collection of the Institute of Biology of the Southern Seas of the Academy of Sciences of the Ukrainian SSR are used in the work. Goldberg optimized environment is prepared for cultivation:
source of nitrogen KNO ₃ - in a concentration of nitrogen 28-42 mg / l;
phosphorus source Na ₂ HPO ₄ - 3-5 mg / l;
trace elements:
iron citrate and perchloric 0.05-0.5 mg / l; CoCl ₃ 0.058 mg / L; MnCl ₂ 0.058 mg / L.

 Carbon dioxide is added in such a way as to maintain a pH of 7.5-8.0.

Culturing is carried out at 23-26 ^{° C,} illuminance of 40 W / m ² FAR, air sparge 1 L per minute in 1 liter of suspension. Creating an optimal growth regime, regulated by the supply of sources of nitrogen, phosphorus and carbon, trace elements, maintaining a certain temperature regime, lighting and sparging contributes to the rapid accumulation of microalgae biomass.

An example implementation of the method. A Nephrochloris salina inoculum (titer 1.0-1.5 × 10 ⁶ cells / ml) was added to the flat cultivator filled with the optimized Goldberg medium, after which the medium was additionally seeded with Platymonas viridis (titer 0.5-1.0 × 10 ⁶ cells / ml ) 2-5 days after the start of microalgae cultivation, the concentration of N. salina reached 22.0 × 10 ⁶ cells / ml, and P. viridis reached 8.7 × 10 ⁶ cells / ml. At these concentrations, a mixed culture exhibits a maximum entomocidal effect.

In a non-intensive culture, the density of N. salina reaches a maximum of 6.0-8.0 × 10 ⁶ cells / ml only at 25-30 days, and P. viridis - 1.0-2.0 × 10 ⁶ cells / ml during this the same period, which indicates the advantage of the proposed method.

 Along with the ability to quickly obtain the required amount of suspension with a certain concentration, intensive continuous cultivation allows you to control and direct the growth process to ensure the desired properties of the culture. The proposed method of cultivation allows you to stabilize the culture at any point of growth, in contrast to non-intensive cultivation.

 PRI me R 1. Leaves of a plane tree were sprayed with a suspension of a mixed culture of N. salina and P.viridis. A caterpillar of an American white butterfly of 2 ages from natural populations was planted on the treated leaves. Three days later, the caterpillars were placed on untreated food, where they continued to feed. After 10-15 days (depending on the stability of the population), as a result of using a suspension of a mixed culture of N. salina and P. viridis, in most of the experiments, 96.7-100% of insects died.

 PRI me R 2. The leaves of the potato were treated with a suspension of mixed culture, and then fed to the larvae of a Colorado potato beetle 2 age. Further, the experiment was carried out according to the method described above. On the 15th day, 90.0% of the larvae died.

 PRI me R 3. Egg-laying of a Colorado potato beetle was sprayed with avolysate of N. salina. On the 10th day, the death of 69.8% of insects was observed. In the future, mortality did not increase.

 The above examples allow us to identify the advantages of the proposed method for producing means of combating insects in comparison with the prototype.

 1. Higher technical efficiency. The death of the caterpillars of the American white butterfly from the P. viridis autolysate does not exceed 91.1% on the 10th day (see auth. St. N 1095483), and insects die from the N. salina autolysate only on the 15th day; continuous mixed culture of the two species leads to almost complete death of the caterpillars on the 10th day.

 2. A wider range of cultural activities. The P.viridis culture is effective mainly against lepidopteran pests, and the effect of the N.salina autolysate on the Colorado potato beetle is inferior to the proposed remedy (examples 2 and 3). The intensive and mixed culture of the two species is highly effective against insects from various systematic groups - the American white butterfly (Lepidoptera) and the Colorado potato beetle (Coleoptera).

 3. High technology. Intensive continuous cultivation techniques make it possible to obtain the required amounts of a working suspension of the required concentration for several days, while it takes 45-60 days to obtain N.salina and P.viridis autolysates with maximum activity; such a prolonged time for the preparation of the active substance makes the practical use of the drug as difficult as the transportation and storage of autolysates.

 The above data show that the method of obtaining a means of combating herbivorous insects due to intensive continuous cultivation of a mixed culture is optimal and can find application in plant protection practice, since it allows you to get a large amount of biomass with high biological activity in a short time.

Claims (1)

 METHOD FOR PRODUCING A MEANS FOR COMBATING VEGETABLE INSECTS, including sowing a culture of algae Nephrochloris salina C. on Goldberg culture medium and cultivating under optimal conditions, characterized in that after sowing a culture of algae Nephrochloris salina C., the medium is additionally inoculated with microalgae R. platonis platis