JP2019147757A - Soil-borne disease mitigation material - Google Patents

Abstract

To provide a novel soil-borne disease mitigation material.SOLUTION: Provided is a soil-borne disease mitigation material that contains three types of bacteria belonging to the genus Bacillus having different characteristics in the carrier material. The three types of bacteria belonging to the genus Bacillus having different characteristics are Bacillus amyloliquefaciens (Accession Number: NITE P-02337), Bacillus subtilis (Accession Number: NITE P-02338), and Bacillus subtilis (Accession Number: NITE P-02354).SELECTED DRAWING: None

Description

  The present invention relates to a soil infectious disease reducing material.

  Various proposals have been made for the purpose of controlling soil-borne diseases.

  For example, in Patent Document 1, a novel filamentous strain capable of infecting and symbiotic to the roots of various plants and having the ability to control soil infectious diseases, a soil infectious disease control material containing cells or cultures of the strains A method for controlling soil infectious diseases of plants using this has been proposed.

  Patent Document 2 has a high control effect on soil infectious diseases caused by phytopathogenic fungi that contain photosynthetic bacteria and Bacillus bacteria as active ingredients, has low environmental impact, is safe and phytotoxic There have been proposed microbial pesticides that are free of soil and methods that can effectively control soil-borne diseases using the microbial pesticides.

  Patent Document 3 proposes a soil infectious disease control method accompanied by crop growth promotion.

JP 2008-67667 A JP2015-39359A Japanese Patent Laying-Open No. 2015-61826

  An object of this invention is to propose a novel soil infectious disease reducing material.

[1]
A soil-borne disease mitigation material that contains three types of Bacillus bacteria with different characteristics in the carrier material.

[2]
The soil infectious disease reducing material according to [1], wherein the carrier material is a porous carrier material having a pore size of μm or more.

[3]
[1] or [2] soil infectious disease reducing material that has been dried to a moisture content of 5 to 10%.

[4]
The soil infectious disease reducing material according to any one of [1] to [3], wherein the bacterial concentration is 1.0 × 10 ⁴ cfu / g to 9.0 × 10 ⁷ cfu / g.

[5]
A soil infectious disease mitigation material that contains three types of Bacillus bacteria with different characteristics in a pre-molded carrier material.

[6]
[5] The soil infectious disease-reducing material according to [5], wherein the carrier material molded in advance is molded into a particle size of 2 mm to 10 mm.

[7]
[5] or [6] soil infectious disease reducing material, wherein the carrier material is a porous carrier material having a pore size of μm or more.

[8]
The soil infectious disease reducing material according to any one of [5] to [7], which is dried to a moisture content of 5 to 10%.

[9]
The soil infectious disease reducing material according to any one of [5] to [8], wherein the bacterial concentration is 1.0 × 10 ⁴ cfu / g to 9.0 × 10 ⁷ cfu / g.

[10]
A soil infectious disease mitigation material formed by molding a carrier material containing three types of Bacillus bacteria with different characteristics.

[11]
[10] soil infectious disease mitigation material molded into a particle size of 2 mm to 10 mm.

[12]
[10] or [11] soil infectious disease reducing material, wherein the carrier material is a porous carrier material having a pore size of μm or more.

[13]
The soil infectious disease reducing material according to any one of [10] to [12], which is dried to a moisture content of 5 to 10%.

[14]
The soil infectious disease reducing material according to any one of [10] to [13], wherein the bacterial concentration is 1.0 × 10 ⁴ cfu / g to 9.0 × 10 ⁷ cfu / g.

[15]
[1] to [14], wherein the carrier material is any one or a combination of dried chicken dung, fermented chicken dung, fermented beef dung, fermented pork dung, diatomaceous earth, zeolite, lightweight cellular concrete, and green pseudo-ashstone. Any soil infectious disease mitigation material.

[16]
The carrier material is a combination of one or more of fermented beef dung, fermented pork, diatomaceous earth, zeolite, lightweight cellular concrete, green pseudo-ashstone, and dried chicken dung and / or fermented chicken dung or dried The soil infectious disease reducing material according to any one of [1] to [14], comprising chicken dung and / or fermented chicken dung.

[17]
Three types of Bacillus bacteria having different characteristics are Bacillus amyloliquefaciens (Accession No .: NITE P-02337), Bacillus subtilis (Accession No .: NITE P-02338), and Bacillus subtilis (Accession No .: NITE P-02354) [1 ] The soil infectious disease reduction material in any one of [16].

  According to this invention, a novel soil infectious disease reducing material can be provided.

The figure which shows the molecular phylogenetic tree by 16S-rRNA gene about Bacillus genus bacteria used for this invention. Reference photograph showing the results of studying the counter-culture of Bacillus amyloliquefaciens (APU-W01), Bacillus subtilis (APU-O02) and Bacillus subtilis (APU-T03) with black root rot causative fungus Calonectria licicola. Reference photos showing the results of examining the response of Bacillus amyloliquefaciens (APU-W01) and Bacillus subtilis (APU-O02) to chitin. The reference photograph showing the result of having examined the influence of the inhibitory bacteria liquid with respect to black root rot by a Pot test. Reference photos showing the results of studying the effects of the application of soil-borne infectious disease mitigation materials on black root rot infection in soybeans. The figure showing the result of having examined the presence or absence of the black root rot infection of the soybean underground part by soil infectious disease mitigation material application. The figure showing the result of having examined the black root rot incidence of soybean by the soil infectious disease reduction material application in each growth period. The figure showing the result of having examined the black root rot onset degree of a green bean by the soil infectious disease reduction material application. The figure showing the result of having examined the influence which the presence or absence of soil infectious disease reduction material application has on the transition of the dry matter weight of soybean.

  Conventionally, Bacillus bacteria have been known to be useful bacteria having properties such as plant growth promotion, disease suppression, and odor reduction.

  The inventors of the present application selected three types of Bacillus bacteria having different characteristics from the Bacillus bacteria and examined the response to soil-borne diseases when applied to plants.

  By this examination, it discovered that the effect which reduces soil infectious disease was exhibited by using simultaneously three types of Bacillus genus bacteria from which a characteristic differs.

  In addition, the effect of reducing soil infectious diseases can be stabilized by adding three types of Bacillus bacteria with different characteristics to the carrier material to reduce soil infectious diseases, thereby reducing soil infectious diseases. It was found that the number of bacteria in the wood and its activity can be stabilized.

  Furthermore, three kinds of Bacillus bacteria having different characteristics are contained in a carrier material and molded into a soil infectious disease reducing material, or three kinds of Bacillus genus having different characteristics are added to a preformed carrier material. By adding bacteria to the soil infectious disease reducing material, the effect of reducing soil infectious disease can be stabilized, and the number of bacteria and the activity in the soil infectious disease reducing material can be stabilized. I found out that I can.

  As the carrier material containing the three types of Bacillus bacteria having different characteristics, a porous carrier material having a minute pore diameter can be used. In order to support three types of Bacillus bacteria having different characteristics, a porous material having a minute pore size is desirable. A porous carrier material having a pore size of μm as the minute pore size, a porous carrier material having a pore size of μm or more, and the like can be employed.

  In the above, the soil infectious disease reducing material can be dried to a moisture content of 5 to 10%. This water content is desirable from the viewpoint of suppressing the growth of other bacteria and stabilizing the number of bacteria and the activity in the soil infectious disease reducing material.

  The carrier material can be any one or a combination of dry chicken dung, fermented chicken dung, fermented beef dung, fermented pork dung, diatomaceous earth, zeolite, lightweight cellular concrete, and green pseudocalcite.

  Or, a combination of any one or more of fermented beef dung, fermented pork, diatomaceous earth, zeolite, lightweight cellular concrete, green pseudo-ashstone, and dried chicken dung and / or fermented chicken dung, or dried chicken dung and / or The carrier material can also be constituted from fermented chicken dung.

  As described above, as a carrier material, a porous carrier material having a minute pore size, for example, a porous carrier material having a pore size of μm units, a porous carrier material having a pore size of μm units or more, etc. When adopting, one or more of diatomaceous earth, zeolite, lightweight cellular concrete and green pseudo-ashstone, which are porous bodies having such minute pore diameters, are used as the material constituting the carrier material. It can be included in one of them.

  In addition, as described later, when Bacillus bacteria isolated from dried chicken manure are used as three kinds of Bacillus bacteria having different characteristics, the growth rate of the three kinds of Bacillus bacteria in a soil infectious disease reducing material From the viewpoint of high manufacturing stress resistance, the carrier material can be constituted from dried chicken dung and / or fermented chicken dung, or can be a carrier material containing at least dried chicken dung and / or fermented chicken dung.

  What was shape | molded using the carrier material mentioned above can be used as the carrier material (= molding material) currently shape | molded. For example, it is possible to use a carrier material (= molding material) that is molded into a particle size of 2 mm to 10 mm.

  In addition to using the carrier material (= molding material) molded as described above, the carrier material described above contains three types of Bacillus bacteria having different characteristics, and is molded into a soil infectious disease reducing material. You can also For example, it can be molded into particles having a particle size of 2 mm to 10 mm.

  Also in this case, it is possible to obtain a soil infectious disease reducing material by drying to a moisture content of 5 to 10%. For example, the molding is performed after drying to a moisture content of 5 to 10%, or after the molding treatment, the moisture treatment is dried to 5 to 10%.

  The carrier material described above contains three types of Bacillus bacteria having different characteristics, or the carrier material formed by molding the carrier material described above, that is, the molded carrier material (= molded material) has a characteristic. By including three different types of Bacillus genus bacteria, or by including three types of Bacillus genus bacteria having different characteristics in the carrier material described above and molding it, the soil infectious disease reducing material of the present invention and can do.

In the above, it is desirable that the bacterial concentration in the soil infectious disease reducing material is 1.0 × 10 ⁴ cfu / g to 9.0 × 10 ⁷ cfu / g.

  As described above, the effect of reducing soil infectious diseases can be stabilized by including three types of Bacillus bacteria having different characteristics in the carrier material. In addition, it is possible to stabilize the number of bacteria and the activity of the soil infectious disease reducing material.

  In including three types of Bacillus bacteria having different characteristics in the carrier material, for example, three types of Bacillus bacteria having different characteristics can be cultured and included in the carrier material.

  For example, three types of Bacillus bacteria having different characteristics are cultured, and a mixed solution obtained by mixing these three types is added to the carrier material in a mass ratio range of 2 to 40%. It is possible to use it as a soil infectious disease reducing material by drying it until it reaches to%.

From the viewpoint of exhibiting the soil infectious disease reducing effect, the soil infectious disease reducing material has a concentration of bacteria of three types of genus Bacillus having different characteristics of 1.0 × 10 ⁴ cfu / g to 9.0 × 10 ^7. cfu / g is desirable.

Therefore, when three types of Bacillus genus bacteria having different characteristics are cultured, a mixture of these three types is mixed and added to the carrier material in a mass ratio of 2 to 40%. It is carried out while adjusting so that the bacterial concentration of three kinds of Bacillus genus bacteria having different characteristics in the infectious disease reducing material is 1.0 × 10 ⁴ cfu / g to 9.0 × 10 ⁷ cfu / g.

In addition, even when three types of Bacillus bacteria having different characteristics cultured are mixed to form a mixed solution, the concentration of the three kinds of Bacillus bacteria having different characteristics in the soil infectious disease reducing material of the final product is 1 Mix while considering to be from 0.0 × 10 ⁴ cfu / g to 9.0 × 10 ⁷ cfu / g. For example, three types of Bacillus bacteria having different characteristics can be mixed at a predetermined mass ratio. As the mixing at a predetermined mass ratio, for example, the above-described three types of Bacillus bacteria having different characteristics can be mixed at an equal mass ratio.

  As one of the soil infectious diseases, soybean black root rot, a soil infectious disease caused by Calonectria ilicicola, is known. Soybean black root rot is a serious problem because no control technology has been established.

  In Odate City, Akita Prefecture, there is a field where soybeans have been continuously cropped for 27 years and have maintained high yields and have little disease such as soybean black root rot. In this field, specific dry chicken manure was applied every year.

  Therefore, the inventors of the present application inferred that there is a microorganism that suppresses soybean black root rot in the dried chicken dung, and tried to isolate a microorganism that suppresses soybean black root rot from the dried chicken dung. By examining the use of isolates, it is possible to reduce soil infectious diseases by simultaneously using three types of Bacillus bacteria having different characteristics, and this invention can be used as a material for reducing soil infectious diseases Was completed.

  In this way, three types of Bacillus bacteria having different characteristics isolated and selected by the inventors of the present application are Bacillus amyloliquefaciens 1 strain and Bacillus subtilis 2 strain. All of these are deposited with the Patent Microorganism Deposit Center of the National Institute of Technology and Evaluation, and have received the following deposit numbers. Bacillus amyloliquefaciens (Accession number: NITE P-02337), Bacillus subtilis (Accession number: NITE P-02338), Bacillus subtilis (Accession number: NITE P-02354).

  A related patent application has already been filed for these three types of Bacillus bacteria (Patent Application Number: Japanese Patent Application No. 2006-191581).

(Classification of strains)
As described above, each of the three strains of Bacillus bacteria having different characteristics selected from the bacteria belonging to the genus Bacillus was identified by decoding the base sequence of the 16S-rRNA gene, and the results shown in FIG. 1 were obtained.

As a result of the 16sRNA analysis, as shown in Table 1, the three strains (Bacillus amyloliquefaciens (accession number: NITE P-02337) and Bacillus subtilis (accession number: NITE P-02338) having an effect on soil-borne diseases are shown. ), Bacillus subtilis (accession number: NITE P-02354)), Bacillus amyloliquefaciens (APU-W01, hereinafter referred to as W-01 in the present specification and drawings), Bacillus subtilis (APU-O02, hereinafter referred to as this), respectively. In the specification and drawings), and Bacillus subtilis (APU-T03, hereinafter referred to as T-03 in the present specification and drawings).

(Adjustment of test strain and inoculum)
The test strains used were W-01, O-02 and T-03. To prepare the bacterial solution, dissolve the LB medium in distilled water, inoculate each single strain into a liquid medium sterilized by autoclaving, and incubate with an incubator at 35 ° C for 3 days while stirring with a stirrer. I went there. The inoculum was prepared by mixing three types of bacterial solutions cultured in a single mixture at the same weight ratio.

(Influence of medium pH)
LB medium and agar were dissolved in distilled water, pH was adjusted to 5.0 to 9.0 with hydrochloric acid and sodium hydroxide, and autoclaved, and then a plate was prepared. Each strain was applied to the prepared plate and cultured at 35 ° C. for 12 hours. The number of colonies after culturing was counted and the proliferative property (actual value / theoretical value) was evaluated. With respect to the growth ability, − indicates no colony formation, − indicates <0.5, + indicates 0.5 ≦ x <1, ++ indicates 1 =, and + indicates +++.

Table 2 shows the evaluation results (proliferation at three different strains at different pHs).

(result)
As a result of evaluating the influence of the medium pH (Table 2), it was shown that the three strains have different growth properties for each pH. In particular, W-01 was highly proliferative on the alkali side at pH 7.0 to 9.0, and T-03 was highly proliferative at a specific pH of 6.0 and 7.0. In addition, O-02 had a characteristic that the pH was 6.0 to 9.0 and the aptitude range was wide. On the other hand, all the bacteria showed low growth at pH 6.0 or lower. From the above results, it was suggested that the growth was high in the range of 6.0 to 6.5, which is the optimum pH for soybean growth, and it was revealed that the responses to the medium pH of the three strains were different.

(Proliferation under low temperature conditions)
After LB medium and agar were dissolved in distilled water (pH 7.8) and sterilized by autoclave, a plate was prepared. The strains were respectively applied to the prepared plates and cultured in the dark at 25 ° C., 15 ° C. and 10 ° C., and the number of days on which colonies were detected was evaluated. As for the growth property under low temperature conditions, those without colony formation were-, those with colonies formed within 2 days of culture were ++, those with colonies formed within 5 days were ++, and colonies were formed within 6 days. The thing was shown as +.

The evaluation results (proliferation properties at low temperatures of the three strains) were as shown in Table 3.

(result)
As a result of examining the growth ability under low temperature conditions (Table 3), colonies were detected in culture within 6 days under the condition of 15 ° C. In particular, O-02 and T-03 were detected within 5 days, indicating the possibility of high sensitivity to low temperatures. In addition, all strains were detected within 2 days under the condition of 25 ° C., indicating that the growth was high. Even when soil infectious disease mitigation materials using three types of strains were sown into the soil at low temperatures, the possibility that the three types of strains could grow in the soil was shown.

(Anti-culture method)
Each strain was inoculated with soybean black root rot fungus on PDA medium, cultured in an incubator at 25 ° C. for 5 days, and examined for the growth inhibitory effect of each isolate on soybean black root rot. The examination result was as shown in the reference photo of FIG. Petri dishes in the photo were inoculated with black root rot fungus on the left side and suppressor or non-inhibitory bacteria on the right side.

(result)
As a result of opposing culture of the three strains and soybean black root rot fungus, it was shown that all the strains had a growth inhibitory effect against black root rot fungus. The degree was estimated to be O-02>W-01> T-03. From the above results, it was shown that the three strains have a growth inhibitory effect on black root rot fungi, and the strength of the three strains suppresses the growth of black root rot fungi by different mechanisms.

(Elucidation of mechanism: response to chitin)
After LB medium and agar were dissolved in distilled water (pH 7.8) and sterilized by autoclave, a plate was prepared. Chitin powder was added to the prepared plate, and each strain was applied. The chitinase activity was estimated by observing whether each strain decomposes chitin added to the medium. The degree to which the inoculated bacterial strain cleared white chitin was evaluated. The examination results were as shown in the reference photo of FIG.

(result)
As a result of estimating the chitinase activity of the strain (FIG. 3), it was shown that all strains degrade chitin in the medium. In particular, the activity of the W-01 strain was remarkably high, and it was shown that O-02 has the activity but the activity may be maintained gently (Date not shown). From the above results, the possibility that any strain having chitinase activity is one of the factors inhibiting the growth of black root rot fungi was shown.

(Pot test)
The test plant was soybean (Glycine max (L.) Merr), and the cultivar was ryuhou. The soil used was sandy loam from Aomori Prefecture, and black root rot fungus was added at a concentration of 10 to the power of 8 to set an infection zone.

  Seeds that were treated with or without inoculation of a suppressive fungus mixed with 3 types of bacteria in the prepared medium (inoculation concentration: 8 article 8 cfu / ml) and surface-sterilized with 70% ethanol and hypochlorous acid Sowing. After sowing, distilled water was added as appropriate in a plant culturing machine, and the mixture was cultured under the conditions of light period / dark period: 28 ° C./18° C. (16 h / 8 h). After the culture, the influence of black root rot was evaluated by observation. The examination results were as shown in the reference photo of FIG.

(result)
As a result of evaluating the effect of inoculation of 3 strains on black root rot in the Pot test (FIG. 4), it was shown that in the group to which the inhibitory bacteria mixed with 3 strains were added, germination and initial growth of soybean were not affected. . It was shown that the concentration of black root rot fungus was high in the group without the inhibitory bacteria solution, but it significantly affected germination and initial growth.

(Examination on materialization)
Fermented chicken dung, lightweight cellular concrete (ALC), and chicken dung carbide were examined as carrier materials. After culturing the three types of Bacillus amyloliquefaciens (APU-W01), Bacillus subtilis (APU-O02), Bacillus subtilis (APU-T03) %, And after molding, heat treatment was performed for 10 hours under the condition of about 60 ° C. (7 kg / batch).

The number of bacteria in the obtained molding material was counted, and the influence of the combination of carrier materials was evaluated. The results were as shown in Table 4 (Study 1 on the combination of carrier materials) and Table 5 (Study 2 on the combination of carrier materials). .

(result)
As a result of the study (Tables 4 and 5), the effect of the combination of materials was hardly recognized, but the tendency to increase the number of bacteria by combining other materials was recognized rather than molding with fermented chicken dung alone . This result is considered to be due to the improved dispersibility of the bacterial solution by adding ALC and green pseudo-ashstone from the value of physical property evaluation of the obtained molding material (date not shown).

(Examination of long-term storage)
After culturing three types of Bacillus amyloliquefaciens (APU-W01), Bacillus subtilis (APU-O02), and Bacillus subtilis (APU-T03), 40% by weight ratio was added to the fermented chicken dung alone, followed by molding and heat treatment. The obtained molding material was cultured in a constant temperature incubator, and the number of bacteria on the 60th day was evaluated.

(result)
As a result of the examination (Table 6), the number of bacteria decreased by about 35% in the 60-day culture test, but it exceeded the fourth power of 10 indicating the reduction effect, and no effect on quality was observed. Became clear.

(Examination of manufacturing stress by large-scale manufacturing machines)
After culturing three types of Bacillus amyloliquefaciens (APU-W01), Bacillus subtilis (APU-O02), and Bacillus subtilis (APU-T03), add 2-10% by weight to the fermented chicken dung alone, and mold with large-scale production equipment Thereafter, heat treatment was performed for 30 minutes at a hot air temperature of about 100 ° C. (800 kg / batch). The number of bacteria of the obtained molding material was evaluated, and the influence of manufacturing stress by a large-scale manufacturing machine was evaluated, and the results were as shown in Table 7 (Influence of manufacturing stress by a large-scale manufacturing machine).

(result)
As a result of the study (Table 7), granulators that are subject to very large stresses such as heat and pressure, and heat treatment, the initial bacterial count is reduced by a factor of about 10 considering the amount of bacterial solution added (about 10% of the mass ratio) However, it was shown that any treatment group can maintain the number of bacteria of 10 4 or more.

  From the above results, it is considered that the stress resistance of the strains used as a carrier for dried chicken manure and fermented chicken manure is extremely high.

  A soil infectious disease mitigation material was applied to the field where plants were grown, and the effect was confirmed.

The test plant was soybean (Glycine max (L.) Merr), and the cultivar was ryuhou. The test field was a rice field conversion field in Aomori Prefecture, and this year was the second field of soybean. The soil type was sandy loam, and soil chemistry (test field chemistry) was as shown in Table 8.

As for the treatment zone, chemical fertilizer is applied as a customary zone (NP ₂ O ₅ -K ₂ O (%): 14-18-14), 200 kg of fermented chicken manure is applied as a carrier zone, and soil infectious disease mitigation material is used as a material zone. 200 kg was applied.

  Three inhibitory strains (W-01, O-02, T-03) were each cultured in LB liquid medium and mixed. 40% of the mass ratio of the fermented chicken dung was added to the obtained mixed liquid, and the molded product was used as a soil infectious disease reducing material. The microorganism molding material was molded into a granular shape and a size of 2-4 mm. The bacterial concentration of the three suppressor strains was 10 5 to 6 cfu / g. Hereinafter, in this example, since it is a molded soil infectious disease reducing material, it is referred to as “microorganism molding material”.

  As for the outline of cultivation, fermented chicken manure was applied to the carrier zone on April 25, microbial molding material was applied to the material zone, and chemical fertilizer was applied to the customary zone on June 6.

  The sowing was performed on June 7 by sowing 72 cm between the ridges, 14 cm between the strains, and 2 grains.

  The survey items were the symptom survey of soil infectious diseases in the subsurface of soybean and the infection rate, growth and yield of soybean black root rot fungus.

  About soil infectious disease symptom investigation, plant body collected in the trilobium (July 8), flowering period (July 29), maximum prosperity period (hereinafter referred to as the most prosperous period) (September 2) The symptom survey of soil infectious diseases (soybean black root rot, soybean stem rot, etc.) was conducted to calculate the severity of the disease.

  Regarding the infection rate, DNA was extracted from soybean roots, and PCR was performed with primers specific to soybean black root rot fungus targeting the β-tubulin gene of filamentous fungi. Then, the infection of soybean black root rot was evaluated by confirming the presence or absence of a band by agarose gel electrophoresis.

  For growth and yield, plants were collected at the trilobal stage, flowering period, and the most prosperous period, and the dry weight was measured.

  On October 14, soybeans were trimmed and the yield and yield components were investigated.

(result)
As a result of evaluating the severity of soil infectious diseases in the soybean subterranean area, the disease level has been lower than the other two test zones from the trilobium to the most prosperous period, and damage caused by the disease has been suppressed. Was shown (FIGS. 5 and 7).

As a result of examining the infection of soybean black root rot fungus, it was shown that the soybean black root rot fungus is not infected in the trilobium stage in the material zone (Fig. 6), and the infection is also observed in the flowering period and the most prosperous period. It was shown that the infection rate was low compared with 2 test plots (Table 9 “Infection rate of soybean black root rot fungus in different growth periods”).

  From the above, it has been clarified that the microbial molding material (soil infectious disease reducing material) has the effect of reducing soybean black root rot. In the trilobium, no infection with soybean black root rot was observed in any of the test plots, but in the material plot, the disease was significantly suppressed compared to the customary plot and the carrier plot (FIG. 7). Therefore, it was estimated that the microbial molding material has an inhibitory effect on soil infectious diseases other than soybean black root rot.

  Edamame test results are shown as reference data. As a result of applying microbial molding materials to supper tea beans and Akitaya Sayaka (200 kg / 10a), the results showed that the disease severity was low compared to the customary area where no materials were applied (FIG. 8).

As a result of investigating the growth, there was no difference in the test weight until the flowering period, but it was suggested that the dry weight was significantly increased in the material group than in the conventional group in the best season (Fig. 9). . As a result of examining the yield, it was shown that the microbial molding material area increased in weight of 100 grains and increased in yield compared to the customary area (Table 10 “Soil infectious disease mitigation agent applied or not) Results of studying the effect on yield ").

  From the above results, microbial molding materials (soil infectious disease mitigation materials) have an inhibitory effect against soybean black root rot fungi, and can reduce or increase the occurrence of disease, thereby increasing or stabilizing soybean growth and yield. It was shown that it can be done. Moreover, it was estimated that it had a mitigation effect also with respect to other soil infectious diseases.

  As mentioned above, although embodiment and the Example of this invention were described, this invention can be variously changed in the technical range grasped | ascertained from description of a claim, without being restricted to these.

  According to this invention, it is possible to make a new proposal for forming a microorganism-shaped molding material having an effect of reducing soil infectious diseases. In particular, it is possible to propose a technique for stabilizing the effect of making a microorganism-molding material having an effect of reducing soil infectious diseases.

  After culturing three types of bacteria belonging to the genus Bacillus having different characteristics, they are mixed at a predetermined mass ratio, for example, an equal mass ratio, and the mass ratio of 2 After encapsulating at a rate of ˜40%, it can be dried to a soil infectious disease reducing material until the water content becomes 5 to 10%.

  The soil infectious disease reducing material proposed by the present invention can maintain the number of bacteria and its activity for 6 months or more even under storage conditions of 25 ° C. or higher. A technique for stabilizing the infectious disease reduction effect can be provided. For example, it is possible to stabilize the effect of reducing the disease against soybean black root rot.

  It has been reported that the causative fungus of soybean black root rot is soil-borne and its persistence is high. Although countermeasures have been taken, chemical pesticides cannot always be treated or are ineffective. Therefore, it is important to stabilize the effect of the strain continuously used as in the present invention, and this technique can be provided.

Claims (17)

  A soil-borne disease mitigation material that contains three types of Bacillus bacteria with different characteristics in the carrier material.   The soil infectious disease reducing material according to claim 1, wherein the carrier material is a porous carrier material having a pore size of not less than µm.   The soil infectious disease reducing material according to claim 1 or 2, which has been dried to a moisture content of 5 to 10%. The soil contagious disease reducing material according to any one of claims 1 to 3, wherein the bacterial concentration is 1.0 x 10 ⁴ cfu / g to 9.0 x 10 ⁷ cfu / g.   A soil infectious disease mitigation material that contains three types of Bacillus bacteria with different characteristics in a pre-molded carrier material.   The soil-borne disease mitigation material according to claim 5, wherein the carrier material molded in advance is molded into a particle size of 2 mm to 10 mm.   The soil infectious disease reducing material according to claim 5 or 6, wherein the carrier material is a porous carrier material having a pore size of μm or more.   The soil infectious disease reducing material according to any one of claims 5 to 7, which has been subjected to a drying treatment to a moisture content of 5 to 10%. The soil contagious disease reducing material according to any one of claims 5 to 8, wherein the fungus concentration is 1.0 x 10 ⁴ cfu / g to 9.0 x 10 ⁷ cfu / g.   A soil infectious disease mitigation material formed by molding a carrier material containing three types of Bacillus bacteria with different characteristics.   The soil infectious disease-reducing material according to claim 10, which is molded into a granular shape having a particle size of 2 mm to 10 mm.   The soil infectious disease reducing material according to claim 10 or 11, wherein the carrier material is a porous carrier material having a pore size of not less than μm.   The soil infectious disease-reducing material according to any one of claims 10 to 12, which has been dried to a moisture content of 5 to 10%. Bacterial concentration is 1.0 * 10 < ⁴ > cfu / g-9.0 * 10 < ⁷ > cfu / g, The soil infectious disease reduction material as described in any one of Claims 10-13.   15. The carrier material according to any one of claims 1 to 14, wherein the carrier material is one or a combination of dried chicken dung, fermented chicken dung, fermented beef dung, fermented pork dung, diatomaceous earth, zeolite, lightweight cellular concrete, and green pseudo-ashstone. The soil infectious disease reducing material according to one item.   The carrier material is a combination of one or a plurality of fermented beef dung, fermented pork, diatomaceous earth, zeolite, lightweight cellular concrete, green pseudo-ashstone, and dried chicken dung and / or fermented chicken dung or dried. The soil infectious disease reducing material according to any one of claims 1 to 14, comprising chicken dung and / or fermented chicken dung.   The three types of Bacillus bacteria having different characteristics are Bacillus amyloliquefaciens (Accession number: NITE P-02337), Bacillus subtilis (Accession number: NITE P-02338), and Bacillus subtilis (Accession number: NITE P-02354). The soil infectious disease reducing material according to any one of 1 to 16.