KR102122748B1 - Long-acting microbial agent for controling meloidogyne spp. in plants and preparing method thereof - Google Patents

Abstract

The present invention relates to a sustained-release microbial preparation for controlling root-knot nematodes, in which antagonistic microbes of root-knot nematodes, Bacilus subtilis and Streptomyces nigrescens, are adsorbed to porous substrates by means of Ecklonia cava Kjellman powder and gelatin, and an extract of Ecklonia cava Kjellman, an extract of Curcuma zedoaria, and an extract of Portulaca oleracea are loaded in the porous substrates and gradually released within soil. In addition, the present invention relates to a method of preparing the sustained-release microbial preparation. The microbial preparation has the antagonistic microbes of root-knot nematodes adsorbed to the substrates and has an extract mixture having insecticidal effects loaded in the substrates, wherein as the antagonistic microbes of root-knot nematodes and the extract mixture are slowly released, not only an excellent effect of controlling root-knot nematodes but also control persistence can be improved.

Description

A sustained-release microbial agent for controlling root-knot nematodes and a manufacturing method therefor {LONG-ACTING MICROBIAL AGENT FOR CONTROLING MELOIDOGYNE SPP. IN PLANTS AND PREPARING METHOD THEREOF}

The present invention relates to a microbial preparation using antagonists of root-knot nematodes, and more specifically, Bacillus subtilis and Streptomyces nigresense, which are antagonists of root-knot nematodes, are adsorbed to porous carriers by persimmon powder and gelatin, and persimmon extract , A mixture of the extract of Achi and the extract of Machihyun is supported on a porous carrier and is slowly released in the soil, and thus relates to a sustained-release microbial preparation for controlling root-knot nematodes and a method for manufacturing the same.

It is known that root knot nematodes not only directly damage plants, but also indirectly allow pathogens such as viruses, bacteria, and fungi to easily invade and produce complex diseases. Meloidogyne nematodes in paper 4 is known as sweet potato root-knot nematode (Meloidogyne incognita), peanut root-knot nematode (M. arenaria), carrot root-knot nematodes (M. javanica) and Java root-knot nematode (M. hapla), of the genus Meloidogyne root-knot nematode Infection creates oxygen radicals that are highly reactive with plant roots, thereby forming toxic intermediates and causing oxidative stress on plants.

Control of root knot nematodes, which negatively affects crop production, as described above, includes chemical control (medicine control against nematodes), physical control (solar heat sterilization, hot water immersion method), and seed control (soil improvement, soil, reptile lubrication, resistant varieties) Various methods such as breeding, resistant crop rotation, manned plant cultivation), and biological control (nematode nematode) have been applied. However, the nematode used for the chemical control has the disadvantage that re-contamination occurs by soil, water, farm equipment, etc., and causes environmental problems such as soil ecosystem imbalance due to high toxicity and long residence period. In particular, as the adverse effects of organic synthetic pesticides on the human body and the natural environment have been revealed, the use of various insecticides has been formulated, and environmentally friendly preparations capable of replacing them are urgently required.

Recently, new insecticidal substances with eco-friendly and specific action mechanisms, such as cultivation of resistant varieties, biological control using natural enemies, and control using plant extracts containing nematode substances to control root-knot nematodes through eco-friendly preparations Research to explore search is ongoing. To date, the ingredients that have a nematode effect include high ginseng extract, wild sesame seed oil, aldehyde, ketones & linolenic acids, and viruses, Rickettsia, invertebrates, bacteria, predatory fungi, parasitic fungi, etc. Has been reported.

Currently, most microbial products sold on the market do not present definite experimental results or are often sprayed on ineffective crops, so they are often not effective, so the development of microbial agents to solve the above problems This is desperate.

Conventionally, Bacillus subtilis KRB-12 (KCTC 18480P) and Streptomyces nigrescens KRA-40 (KCTC 18482P) are known as microorganisms for controlling root-knot nematodes. The microorganisms are effective in decomposing the nematode of the root knot nematode in a short time or decomposing the nematode, but no method for effectively controlling the root knot nematode for a long time is known, and a formulation technique for improving the effect persistence for a long time is also known. none.

Gamtae grows on a lunch table about 10 m deep and grows up to 1 to 2 m long. The stem is cylindrical, and the root is root-shaped, and at the end of the stem, there is one flat middle leaf with a side leaf fragment. This leaf is about 1 m long and brown, but when it dries, it becomes black, thick, leathery, and has small leaves on the sides. Persimmon is an important algae plant that makes up the marine forest, and mainly feeds abalone and conch. It is a major raw material for making alginic acid, iodine, and potassium, such as brown seaweed such as seaweed and kelp, and can be collected and used for food, but is rarely consumed. In Korea, it is mainly distributed in the south coast and Jeju. Recently, it is known about sterilization or insecticidal effect of persimmon and its use for adsorbing heavy metals, but there is no known insecticidal effect for root-knot nematodes or its use for adsorbing microorganisms to carriers.

Achul is a dried stem of the bud of Gingeraceae, and has been traditionally used for indigestion, gout, colic, and menstruation, and is known to have hepatocyte protection. In addition, it is known to be effective in controlling anthrax anthrax, and has macrophage activation ability and pancreatic lipolytic inhibitory activity. However, the insecticidal effect against root-knot nematodes is not known.

Machihyeon has been used extensively as a medicine because it has excellent efficacy in treating skin diseases such as boils and edema without irritating it with a perennial plant of biscuit plant. Recently, it has a skin moisturizing effect and a skin whitening effect as well as a skin disease, and has been used as a functional cosmetic composition. However, the effect of insecticidal effects on root-knot nematodes is also unknown.

Thus, the present inventors have overcome the problems of the prior arts and can effectively control the root-knot nematode, and as a result of earnest research to implement a sustained-release preparation for maintaining the control effect for a long time, Bacillus subtilis, an antagonist of the root-knot nematode KRB-12 and Streptomyces nigresense KRA-40 are adsorbed on a porous carrier by persimmon powder and gelatin, and in the case of sustained-release microbial preparations for root or nematode control in which persimmon extract, chul extract and marchi string extract are supported on the porous carrier In addition, it can effectively decompose the root-knot nematode or the ovary of the root-knot nematode as well as the antagonistic bacteria adsorbed on the carrier by persimmon and gelatin, and the mixed liquor, effusion and guchihyun extracts supported on the carrier, thereby gradually maintaining the control effect. It confirmed, and completed this invention.

Therefore, the main object of the present invention is not only to effectively decompose the root-knot nematode or the ovary of the root-knot nematode, but also the antagonistic bacteria adsorbed on the carrier by persimmon and gelatin, and the mixed solution of sensitized ecstasy, ichthyosis and guchi-hyun extracts slowly released. It is to provide a sustained-release microbial preparation for root-knot nematode control that can sustain the.

Another object of the present invention is to provide a method for preparing a sustained-release microbial agent for controlling root-knot nematodes for preparing the sustained-release microbial agent for controlling the root-knot nematode.

According to one aspect of the present invention, in the present invention, in a microbial preparation using an antagonist of a root-knot nematode, Bacillus subtilis and Streptomyces nigresense, which are antagonists of a root-knot nematode, are adsorbed to a porous carrier by persimmon powder and gelatin. It provides a sustained-release microbial preparation for root-knot nematode control, characterized in that the persimmon extract, chul extract and machihyeon extract are supported on a porous carrier and gradually released in the soil.

Conventionally, in order to control root-knot nematodes through eco-friendly preparations, research has been conducted on methods of controlling resistant varieties using cultivation of resistant varieties, biological control using natural enemies, and plant extracts containing nematode substances. Methods to improve the lasting effects for nematode control have not been studied. Thus, the present inventor adsorbs the antagonists against the root-knot nematodes to the carrier, and when the extract mixture having the insecticidal effect is supported on the carrier to prepare a microbial agent, the control persistence is improved as the antagonists and extracts against the root-knot nematodes are slowly released. It confirmed, and completed this invention.

According to one experimental example of the present invention, as a result of selecting suitable adsorbents for adsorbing microorganisms to carriers from various adsorbents known as biodegradability, it was confirmed that persimmon and gelatin are the best in viscosity, dissolution and clumping properties (Experimental Example 5) And Table 6), and the present inventors tried to improve the adsorption effect of microorganisms by mixing a gelatin with gelatin with not only adsorption but also insecticidal effect.

According to an experimental example of the present invention, in the case of a microbial preparation prepared by mixing the above feelings and gelatin, it was confirmed that the smallest amount of microorganisms is released even after 24 hours have elapsed. These results suggest that the microbial formulation of the present invention using persimmon and gelatin as a biodegradable adsorbent is suitable as a sustained release formulation.

According to an experimental example of the present invention, in the case of sustained-release microbial preparations prepared by using persimmon extract and gelatin as adsorbents, the persimmon extract, chul extract and guchihyeon extract exhibit excellent effects in terms of control effect and duration of effect. It was confirmed (see Experimental Example 7 and Table 10). These results suggest that the sustained-release microbial preparation for controlling root-knot nematodes according to the present invention is suitable to effectively decompose and kill root-knot nematodes or oocytes for a long time.

In the microbial preparation according to the present invention, the Bacillus subtilis is Bacillus subtilis KRB-12 ( Bacillus subtilis KRB-12; KCTC 18480P), and the Streptomyces nigresense is Streptomyces nigresense KRA-40 ( Streptomyces nigrescens KRA-40; KCTC 18482P).

In the microbial preparation according to the present invention, the persimmon powder and gelatin can be mixed in any ratio having viscosity, and is preferably characterized by being mixed at a ratio of 20 to 30:60 to 70. When the ratio of the gelatin is 60 or less, the viscosity may be weak and the adsorption power may be lowered. When the ratio of the gelatin is 70 or more, the viscosity may be too strong, and the release of microorganisms may be reduced. For example, when 20 to 30 g of persimmon powder is included, gelatin may be mixed to include 60 to 70 g.

In the microbial preparation according to the present invention, the porous carrier can support the extract mixture, and any carrier formed with porosity can be used, preferably selected from the group consisting of diatomaceous earth, zeolite, maculite, vermiculite, sand and activated carbon. It may be one or more carriers, more preferably characterized in that the diatomaceous earth.

In the microbial preparation according to the present invention, the diatomaceous earth has a particle size of 20 to 40 mesh, and a specific surface area of 10 to 14 m 2 /g.

According to another aspect of the invention, the present invention is (a) Bacillus subtilis KRB-12 ( Bacillus subtilis KRB-12; KCTC18480P) strain and Streptomyces nigrescens KRA-40; KCTC 18482P) culturing the strain in a liquid medium to prepare a microbial culture solution; (B) preparing the adsorbent solution by mixing persimmon powder, gelatin and distilled water; (c) preparing an extract mixture by mixing persimmon extract, chul extract and machi extract; (d) mixing the microbial culture solution of step (a), the adsorbent lysis solution of step (b) and the extraction mixture solution of step (c) and spraying the mixture onto a carrier; (e) stirring the carrier of step (d) in the microbial culture solution of step (a), the adsorbent lysis solution of step (b) and the extraction mixture of step (c); And (f) drying the carrier of step (e); provides a method for preparing a sustained-release microbial agent for controlling root-knot nematodes.

In the method for preparing a sustained-release microbial agent for controlling root-knot nematodes of the present invention, the liquid medium of step (a) can be any liquid medium capable of culturing the root-knot nematode antagonist, preferably corn starch, ( NH4) ₂ SO ₄ and With KH ₂ PO ₄ It is characterized by being configured.

In the method for preparing a sustained-release microbial agent for controlling root-knot nematodes of the present invention, the adsorbent lysate in step (b) is mixed with persimmon powder and gelatin in a ratio of 20 to 30: 60 to 70, and the concentration of sorbent lysate is 0.75. % It is characterized in that it is prepared by putting it in distilled water and heating it. When the ratio of the gelatin is 60 or less, the viscosity may be weak and the adsorption power may be lowered. When the ratio of the gelatin is 70 or more, the viscosity may be too strong, and the release of microorganisms may be reduced. In addition, when the concentration is 0.75% or less, the viscosity may be low and the adsorption power may be lowered. When the concentration is 0.75% or more, the viscosity may be too strong, and thus the release of microorganisms may be reduced.

According to an experimental example of the present invention, when the concentration of the biodegradable adsorbent is 0.75%, it was confirmed that it exhibits better sustained release than when it is 0.75% or less (see Experimental Example 6 and Table 9). These results suggest that the concentration of biodegradable adsorbents is an important component in preparing sustained-release microbial preparations.

In the method for preparing a sustained-release microbial preparation for controlling root-knot nematodes of the present invention, the extract mixture of step (c) is characterized by mixing after extracting persimmons, shoots, and gusset, respectively. The persimmon extract: Achul extract: Machihyeon extract may be mixed in a weight ratio of 1 to 2: 0.5 to 1.5: 0.1 to 0.3, and more preferably 1.5 to 1: 0.2 in weight ratio.

The extracts of persimmon, ichthy and marchi of the present invention can be extracted and separated according to conventional methods known in the art, that is, under the conditions of conventional temperature and pressure, using a conventional solvent. As an extraction solvent for extracting the persimmon, achur, and marchi extract of the present invention, a solvent that can be generally used in the extraction process may be used, preferably, water, anhydrous or hydrous alcohol having 1 to 6 carbon atoms, hexane (hexane) ), acetone, ethyl acetate, butyl acetate, 1,3-butylene glycol or a combination thereof, but is not limited thereto.

In the method for preparing a sustained-release microbial preparation for controlling root-knot nematodes of the present invention, the drying in step (f) is characterized in that the carrier in step (e) is stirred at a temperature of 20 to 30°C to dry moisture.

As described above, the sustained-release microbial preparations for controlling root-knot nematodes according to the present invention can not only effectively decompose the root-knot nematode or the oocyte of the root-knot nematode, but also the antagonistic bacteria adsorbed on the carrier by gelatin and gelatin, and the emotional state supported on the carrier, The control solution can be continuously maintained by slowly discharging the mixed liquid of extracts from Achu and Machi.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Preparation Examples 1 and 2: Microorganism control against root-knot nematodes and preparation of culture broth

In the present invention, Bacillus subtilis KRB-12 (KCTC18480P) strain and Streptomyces nigrescens KRA-40; KCTC 18482P as microorganisms for decomposing root-knot nematode oocytes and nematodes ) Strains were selected. The selected strains were cultured for 3 days at 37°C in a liquid medium composed of Table 1 below to prepare a microbial culture solution for controlling root-knot nematodes.

Culture composition content Corn starch One% (NH4) ₂ SO ₄ 0.09% KH ₂ PO ₄ 0.01%

Experimental Example 1: Confirmation of the control effect of root-knot nematodes (1)

Using the microbial culture medium (Bacillus subtilis KRB-12) prepared in Preparation Example 1, the activity of decomposing the follicle nematodes was measured by measuring the change in the number of soil and root nodules through pot experiments. It was confirmed, and the results are shown in Table 2 below.

Treatment Day 1 of inoculation Day 30 of inoculation Day 60 Number of nematodes Per 100 g of soil
Number of caterpillars Per root
Number of ovaries Number of caterpillars per 100 g of soil Per root
Number of ovaries H ₂ O - - - - - LB culture 1030±67.08 1306±82.95 78.8±6.53 1830±69.28 140.8±10.06 Preparation Example 1 1034±63.87 582±107.33 53.2±7.85 1108±98.59 73.8±6.98

As a result, as can be seen in Table 2, it can be seen that the microbial culture medium of Preparation Example 1 significantly reduced the number of oocytes than the control LB culture medium.

Experimental Example 2: Confirmation of control effect of root-knot nematodes (2)

Using the microbial culture medium prepared in Preparation Example 2 (Streptomyces nigresense KRA-40), the potency of the root knot nematode sac targeting pepper seedlings was measured through a pot experiment to measure changes in soil and root knot counts. Degradation activity was confirmed, and the results are shown in Table 3 below.

Treatment Day 1 of inoculation Day 30 of inoculation Day 60 Number of nematodes Per 100 g of soil
Number of caterpillars Per root
Number of ovaries Per 100 g of soil
Number of caterpillars Per root
Number of ovaries H ₂ O - - - - - GSS culture 1012±63.01 1300±72.1 79.2±6.83 1812±65.35 152±8.15 Preparation Example 2 1030±51.48 366±60.25 30.2±8.41 538±44.38 40.0±7.31

As a result, as shown in Table 3, it can be seen that the microbial culture medium of Preparation Example 2 significantly reduced the number of oocytes than the control GSS culture medium.

Experimental Example 3: Confirming the control effect of root-knot nematodes (3)

In order to confirm the root worm nematode control effect of persimmon, the change of soil and root nodules was measured through pot experiments on the decomposition activity of the root worm nematode using pepper persimmon extract, extract, and Machihyun extract in distilled water. Oocyte decomposition activity was confirmed, and the results are shown in Table 4 below.

Treatment Day 1 of inoculation Day 30 of inoculation Day 60 Number of nematodes Number of caterpillars per 100 g of soil Per root
Number of ovaries Number of caterpillars per 100 g of soil Per root
Number of ovaries H ₂ O - - - - - Persimmon extract 1050±49.48 381±40.44 41.1±8.70 555±33.38 58.0±1.21 Achul extract 1045±93.26 375±81.54 39.9±5.57 543±10.01 57.1±9.17 Machi Prefecture Extract 1047±51.26 377±52.24 40.2±9.51 534±64.43 56.6±5.46

As a result, as can be seen in Table 4, the persimmon mixed solution, the extract of ichthym and the extract of Machihyun had fewer fewer ovarian follicles than the microbial culture solution of Preparation Example 1 and Preparation Example 2, but the effect of decomposing the oocyst to be said to have the effect of decomposing the oocyst It was shown. Through these effects, it was confirmed that the Ecklonia cava extract, Achul extract and Machihyeon extract can be used as components for controlling root-knot nematodes.

Experimental Example 4: Selection of carrier

In order to prepare a microbial agent for controlling root-knot nematodes in a granular form, carriers for adsorbing or supporting microorganisms were investigated, and carriers of five candidates were compared, and the results are shown in Table 5 below.

division Affinity Physical strength Granularity Diatomaceous earth ◎ ◎ ◎ Activated carbon ◎ ○ ○ sand ○ △ × Vermiculite ○ × △ Fulvic acid △ × ×

◎: Very good, ○: Excellent, △: Normal, ×: Poor

As a result, as shown in Table 5, diatomaceous earth was found to be most suitable as a carrier in terms of affinity, physical strength, and particle size with microorganisms.

Experimental Example 5: Selection of biodegradable adsorbent

The material as an adsorbent was investigated to adsorb microorganisms to a carrier and elute in a sustained release manner. Among them, the adsorbents of five candidates were compared, and the results are shown in Table 6 below.

division Solubility viscosity Bunch Feeling ○ ◎ ◎ gelatin ○ ○ ○ Starch △ ○ ○ PVB × × × PLA × × △

◎: Very good, ○: Excellent, △: Normal, ×: Poor

As a result, as can be seen in Table 6, it can be seen that persimmon is most suitable for the solubility in water, viscosity in water, and agglomeration during dissolution and formulation. In addition, it was judged to be suitable as an adsorbent because it is not superior to persimmon even in the case of gelatin, but is excellent overall. Accordingly, the present inventors selected persimmon and gelatin as adsorbents.

Example 1: Preparation of sustained release microbial preparation (1)

A sustained-release microbial preparation was prepared by using gluten and gelatin, which are the adsorbents selected in Experimental Example 4, as an adhesive. The specific method is as follows.

First, microbial cultures are prepared according to the manufacturing methods of Preparation Examples 1 and 2. Then, the sensitizer powder and gelatin, which are adsorbents, were mixed in a ratio of 30:70, placed in distilled water so that the concentration was 0.75%, and heated at 70 to 90°C for 1 to 2 hours to completely dissolve the gluten powder and gelatin, and then cooled at room temperature. A biodegradable adsorbent lysate is prepared. The prepared microbial culture solution and the adsorbent lysis solution are mixed and then evenly adsorbed by spraying the diatomaceous earth as a carrier by a spray method. After spraying was completed by spraying, diatomaceous earth adsorbed by microorganisms was supported in a mixed solution of microbial culture solution and adsorbent soluble solution to increase adsorption and dispersibility while stirring. The sustained-release microbial preparation was prepared by drying the moisture in the mixture by stirring the adsorbed carrier at a temperature of 20 to 30°C.

Comparative Examples 1 to 3: Preparation of sustained-release microbial preparations

In order to confirm the sustained-release efficacy according to the type of adsorbent, a microbial preparation was prepared in the same manner and in the same manner as in Example #1 as a comparative example, but persimmon or gelatin was used as an adsorbent, respectively, or different types of adsorbent as shown in Table 7 below. Microbial preparation was prepared using.

Comparative Example 1 Persimmon powder Comparative Example 2 gelatin Comparative Example 3 PVB

Experimental Example 5: Confirmation of sustained release effect according to the type of biodegradable adsorbent

The microbial preparations prepared according to Examples 1 and 1 to 3 were measured to measure the density of microorganisms over time to confirm the sustained-release effect according to the type of biodegradable adsorbent, and the results are shown in Table 8. Did.

Number of initial spores
(×10 ⁸ cfu/g) Released after 24 hours
Number of spores (×10 ⁸ cfu/g) Released 24 hours
Percentage of spores (%) Example 1 (Persimmon powder + gelatin) 3.5 0.21 6 Comparative Example 1 (Kim Tae Powder) 4.5 1.15 25.6 Comparative Example 2 (gelatin) 3.5 0.4 11.4 Comparative Example 3 (PVB) 3.5 0.8 22.8

As a result, as can be seen in Table 8, it can be seen that the microbial agent prepared in Example 1 of the present invention released the smallest amount of microorganisms even after 24 hours have elapsed. In particular, in the case of Comparative Example 1, although it showed the highest microbial release rate, it can be seen that when used in combination with gelatin, it showed the lowest microbial release rate.

Experimental Example 6: Confirmation of sustained release effect according to the concentration of biodegradable adsorbent

For the microbial preparation prepared according to Example 1, the density of microorganisms over time was measured to confirm the sustained-release effect according to the concentration of the biodegradable adsorbent, and the results are shown in Table 9. The manufacturing method was the same as in Example 1, but the concentration of the biodegradable adsorbent was different to prepare a microbial preparation.

Concentration of Example 1 (%) Number of initial spores
(×10 ⁸ cfu/g) Released after 24 hours
Number of spores (×10 ⁸ cfu/g) Released 24 hours
Percentage of spores (%) 0.5 3.0 0.3 10 0.75 3.5 0.21 6 1.0 3.7 0.67 19.1

As a result, as can be seen in Table 9 above, it exhibits excellent sustained release at all concentrations, but it can be seen that when the concentration of the biodegradable adsorbent is 0.75%, it exhibits the best sustained release.

Example 2: Preparation of sustained release microbial preparation (2)

A sustained-release microbial preparation was prepared by using the Ecklonia cava extract, the Achi extract and the Machihyun extract, in which the root-knot nematode control effect was confirmed in Experimental Example 3, and the Ecstasy and Gelatin, the adsorbent selected in Experimental Example 4, as an adhesive. The specific method is as follows.

First, microbial cultures are prepared according to the manufacturing methods of Preparation Examples 1 and 2. Then, the sensitizer powder and gelatin, which are adsorbents, were mixed in a ratio of 30:70, placed in distilled water so that the concentration was 0.75%, and heated at 70 to 90°C for 1 to 2 hours to completely dissolve the gluten powder and gelatin, and then cooled at room temperature. A biodegradable adsorbent lysate is prepared. In another mixing container, extracts of persimmon extract, chul extract and machihyeon extract are prepared to prepare an extract mixture. The prepared microbial culture solution, adsorbent lysate and extraction mixture are mixed and then evenly adsorbed by spraying the diatomaceous earth as a carrier in a spray manner. After spraying was completed by spraying, diatomaceous earth adsorbed by microorganisms was supported in a mixed solution of a microbial culture solution, an adsorbent lysate, and an extract mixture to increase adsorption and dispersibility while stirring. The sustained-release microbial preparation was prepared by drying the moisture in the mixture by stirring the adsorbed carrier at a temperature of 20 to 30°C.

Experimental Example 7: Evaluation of control effect and duration of effect

In order to evaluate the control effect and duration of effect of the microbial agent according to Example 2 of the present invention, the soil of the red pepper arable land, which had severe root-knot nematodes in the previous year, was secured and a simple pot was produced using the same (61×40×15cm) After that, the pepper was grown on a small scale to evaluate the control and duration of effect of the microbial agent of the present invention, and the results are shown in Table 10 below. Control was expressed as a percentage of the whole pepper seedlings of the test. The growth of the above-ground part was given as 1 point in case of poor quality, 2 points in case of a slight amount, 3 points in case of normal, 4 points in case of good, and 5 points in case of very good.

Cultivation days Treatment Total pepper seedlings Incidence rate (%) Control (%) Above-ground growth 10 days Example 2 50 0 - 5 pesticide 50 0 - 5 No treatment 50 0 - 5 20 days Example #2 50 2.9 - 4.5 pesticide 50 0 - 4.5 No treatment 50 23.0 - 3 30 days Example 2 50 5.5 - 4 pesticide 50 5.5 - 4 No treatment 50 50.7 - 3 40 days Example 2 50 9.7 - 4 pesticide 50 10.5 - 4 No treatment 50 69.7 - 2.5 50 days Example 2 50 14.7 85.3 3.5 pesticide 50 23.2 76.8 3.5 No treatment 50 79.8 20.2 1.5 60 days Example 2 50 18.7 81.3 3.5 pesticide 50 31.2 68.8 3.5 No treatment 50 88.2 11.8 One

As a result, as can be seen in Table 10, the microbial preparation of Example 2 according to the present invention can sustain the control effect on the root-knot nematodes for a long time because the extracts for controlling antagonists and root-knot nematodes of the root-knot nematodes are slowly released.

Claims (10)

In the microbial preparation using root antagonist antagonists,
Bacillus subtilis and Streptomyces nigresense, which are antagonists of the root-knot nematodes, are adsorbed on the porous carrier by persimmon powder and gelatin, and persimmon extract, chul extract and marchi string extract are supported on the porous carrier and slowly released from the soil. A sustained-release microbial agent for controlling root-knot nematodes.
According to claim 1,
The Bacillus subtilis is Bacilus subtilis KRB-12 (KCTC 18480P), and the Streptomyces nigresense is Streptomyces nigrescens KRA-40; KCTC 18482P It characterized in that the sustained-release microbial preparations for controlling root-knot nematodes.
According to claim 1,
The persimmon powder and gelatin is a sustained-release microbial preparation for controlling root-knot nematodes, characterized in that it is mixed at a ratio of 20 to 30: 60 to 70.
According to claim 1,
The porous carrier is a sustained-release microbial agent for controlling root-knot nematodes, characterized in that it is at least one carrier selected from the group consisting of diatomaceous earth, zeolite, maculite, vermiculite, sand and activated carbon.
The method of claim 4,
The diatomaceous earth has a particle size of 20 to 40 mesh (mesh), and a specific surface area of 10 to 14 m 2 /g.
Method for preparing a sustained-release microbial preparation for controlling root-knot nematodes comprising the following steps:
(a) Bacillus subtilis KRB-12 (Bacillus subtilis KRB-12; KCTC18480P) strain and Streptomyces nigresense KRA-40 (Streptomyces nigrescens KRA-40; KCTC 18482P) strain were cultured in a liquid medium to incubate the microbial culture medium. Manufacturing;
(B) preparing the adsorbent solution by mixing persimmon powder, gelatin and distilled water;
(c) preparing an extract mixture by mixing persimmon extract, chul extract and machi extract;
(d) mixing the microbial culture solution of step (a), the adsorbent lysis solution of step (b) and the extraction mixture solution of step (c) and spraying the mixture onto a carrier;
(e) stirring the carrier of step (d) in the microbial culture solution of step (a), the adsorbent lysis solution of step (b) and the extraction mixture of step (c); And
(f) drying the carrier of step (e).
The method of claim 6,
The liquid medium of step (a) is corn starch, (NH4) ₂ SO ₄ and With KH ₂ PO ₄ Method for producing a sustained-release microbial preparation for controlling root-knot nematodes, characterized in that configured.
The method of claim 6,
The sorbent lysate in step (b) is prepared by mixing persimmon powder and gelatin in a ratio of 20 to 30: 60 to 70, and putting it in distilled water so that the concentration is 0.75%, and heating it. Method of manufacturing.
The method of claim 6,
Method of producing a sustained-release microbial preparation for root-knot nematode control, characterized in that the extract mixture of step (c) is extracted after each of persimmon, shoot, and marchi.
The method of claim 6,
The drying of step (f) is a method of preparing a sustained-release microbial agent for controlling root-knot nematodes, characterized in that the carrier of step (e) is dried at 20 to 30° C. to dry moisture.