JPH0791141B2 - Soil disease control material and soil disease control method using the same - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a control material and a control method for soil diseases which are difficult to control.

[0002]

2. Description of the Related Art Rotation of a cropping system has been a conventional technique for avoiding soil diseases. However, in recent years, the traditional crop rotation system has collapsed due to selective expansion of highly profitable crops such as vegetables and a vegetable production area designation system, and continuous cropping of single crops has become common. For this reason, soil diseases frequently occur nationwide, and as a countermeasure, large amounts of pesticides are inevitably used.

[0003]

The fumigation and sterilization of soil by pesticides poses a problem such as danger to human body in handling, necessity of coating material such as vinyl, and adverse effect on human livestock due to gas volatilization to the surrounding area. ing. In addition, the steam sterilization method and the hot water sterilization method are used only in some of the horticultural facilities in terms of equipment cost. Further, there is a method of controlling by applying crab, krill, shrimp or the like to the soil, but it is reported that the effect is not stable and the disease may be accelerated. On the other hand, recently, biological control using an antagonistic microorganism has been attempted, but it is difficult to fix a specific microorganism in soil for a long time, and a sufficient effect has not been achieved.

That is, an object of the present invention is to provide a soil disease control material capable of allowing antagonistic actinomycetes to settle in soil for a long period of time, and a soil disease control method using the same.

[0005]

Means for Solving the Problems The present invention provides a fermented product obtained by fermenting a chitin or a chitin-containing organic substance and a plant organic substance, zeolite, and Streptomyces anuratus, which is an antagonistic actinomycete capable of producing chitinase. The present invention relates to a soil disease control material containing the same. Furthermore, the present invention, Streptomyces anuratus is an antagonistic actinomycete having a chitinase-producing ability, the ratio of the fermented product and zeolite obtained by fermenting chitin or a chitin-containing organic matter and a plant organic matter in a weight ratio of 99. The present invention relates to a soil disease control material obtained by proliferating in a culture medium of 1 to 1:99.

Further, the present invention is a method for controlling soil diseases, which comprises applying a soil disease controlling material to soil. The chitin used in the present invention is chitin obtained by decalcifying crab or lobster by acid treatment and deproteinizing by alkali treatment, and may be in the form of flakes or powder. Chitin is a chitin-containing organic substance,
For example, powders of crab, shrimp, krill, etc. may be substituted.

As the vegetable organic matter, rice bran, defatted rice bran, bran, rapeseed, rice straw, etc. are preferred, and the combination of defatted rice bran and bran is particularly advantageous. To obtain a fermented product by fermenting the chitin or chitin-containing organic matter and plant organic matter, for example, shellfish powder, defatted rice bran,
After mixing each raw material from bran etc., add water and add 30
~ There is a method of fermentation for 40 days.

[0008] The quality of the zeolite used in the present invention is not particularly limited, and an agricultural zeolite which is usually used as a soil improving material is sufficient. The ratio of the fermented product to the zeolite is preferably 99: 1 to 1:99, and more preferably 2: 8 to 1: 9 by weight. In the present invention, Streptomyces anulatus C which is an antagonistic actinomycete having a chitinase-producing ability as a microorganism
KJ (Streptomyces sp. CK-J) (this microorganism has been deposited at the Institute of Microbial Science and Technology of the Institute of Industrial Science and Technology as Microorganism Research Institute No. 11783). This actinomycete is isolated from the soil to which chitin or an organic substance containing chitin is applied, and produces a highly active chitinase and an antibiotic having a broad antibacterial spectrum.

[0009] Streptomyces anuratus CK-J has the following mycological properties. (1) Cell wall type I LL- diaminopimelic acid + meso-diaminopimelic acid - diaminobutyric acid - glycine + aspartic acid - ornithine - lysine - arabinose - galactose - (2) quinone _{MK-9 (H 6),} MK-9
(H ₈ ) (3) Presence of aerial hyphae (4) Morphology of spore chains Linear to curved (5) Surface structure of spores Smooth (6) Color of the surface of the community Yellow (7) Production of melanin pigment + (8) Color of the backside of the village Yellow (9) Production of water-soluble dye − (10) Assimilation Arabinose − Xylose + Inositol − Mannitol + Fructose + Rhamnose + Sucrose − Raffinose − (11) Non-motile (12) ) No sporangia (13) Grow well in a medium containing chitin as the sole carbon source. (14) It shows antibacterial activity against Fusarium bacterium, Rhizoctria bacterium, Verticillium bacterium, etc.

In the present invention, Streptomyces sp. CK-A, which is an antagonistic actinomycete capable of producing chitinase as another microorganism, is contained in the Institute of Microbial Science and Technology, Institute of Industrial Science and Technology, Microbiology Research Institute No. 11413. Has been deposited as)
Can be used. This Streptomyces sp.CK-A
Is an actinomycete isolated from soil containing chitin and secretes several chitinases with high chitin-degrading activity. The secreted chitinase includes endo-type and exo-type, efficiently decomposes chitin, and mainly produces a mixture of chitopentose from chitotriose.

The soil disease controlling material of the present invention can effectively control soil diseases when applied to soil. In the present invention, the target soil diseases are tomato wilt, tomato root rot, potato brown rot, spinach wilt, strawberry yellow rot, yugao wilt disease, watermelon wilt disease, apple white wilt disease, apple. Examples include purple crotch rot, apple rot rot, eggplant half-rot rot, potato root rot, Japanese radish seedling death, and Rhizoctonia large patch, which is a disease of turf.

[0012]

EXAMPLES The present invention will now be described in detail with reference to examples, but the present invention is not limited to these examples. Reference Example 1 Fusarium oxysporum f.sp fragaria
The cell wall of e ) was prepared and an agar plate medium was prepared using this as a carbon source. When Streptomyces anuratus CK-J was cultured in this medium, a large zona pellucida was formed around the bacterial cells, confirming the filamentous fungal cell wall degrading ability of this microorganism. Further, when cultured in a liquid medium containing the above-mentioned carbon source, a high level of chitinase activity was observed in the culture supernatant.

Reference Example 2 Streptomyces anuratus CK-J was added to chitin medium (Czape
medium containing 12 g of chitin powder to 4.01 g / L of inorganic salt of k)
The chitinase accumulated in the culture filtrate was measured as the specific activity per protein. The quantification of chitinase activity in the culture filtrate was carried out by measuring the reducing sugar using glycol chitin as a substrate [T. Imoto, K. Yagishita, Agric. Biol. Che.
m., 35 , 1154 (1971)].

As a result, the highest activity was observed on the 7th to 9th day of culture, and the value was 4 to 5 times that of the commercially available chitinase derived from Streptomyces griseus. Reference Example 3 Table 1 shows the results of confrontation culture of Streptomyces anuratus CK-J and various soil pathogens in potato dextrose agar medium.

[0015]

[Table 1]

Preparation Example 1 The types and mixing ratios of raw materials are shown below. As the chitin-containing organic substance, crab mosquitoes are used, but shrimp moss may be used.
As the vegetable organic matter, defatted rice bran, bran, etc. are used. Vermiculite and zeolite are used as mineral substances. The mixing ratio of each material is crab,
30-50% by weight of chitin-containing organic matter such as shrimp, 60-80% by weight of vegetative organic matter such as defatted rice bran, bran, tofu residue, and 5-20% by weight of mineral substances such as vermiculite and zeolite good.

These raw materials are added to a fermenter, mixed and then added with water of the same weight, and fermented in the fermentor for 30 to 40 days to complete ripening. Example 1 A method for efficiently growing Streptomyces anuratus CK-J in the fermented product obtained in Preparation Example 1 (hereinafter referred to as “chitin-containing fermented organic matter”) was examined. Mineral substances such as zeolite and vermiculite were mixed with the chitin-containing fermented organic matter, and the present microorganism was cultured. As a result, when zeolite was mixed, the growth was the fastest and the bacterial density in the material was high.
For example, when chitin-containing fermented organic matter and zeolite are mixed at a weight ratio of 1: 1 and 10 ⁴ cfu of this microorganism is inoculated per 1 g of the material, 10 ⁹ cfu of this microorganism per 1 g of the material is obtained after 10 days.
Became. This test was performed in an open system with non-sterile materials. Therefore, it can be said that the above-mentioned culture method is very excellent as a large-scale culture method for a specific antagonistic microorganism.

Example 2 The mixture ratio of chitin-containing fermented organic matter and zeolite most suitable for the growth of Streptomyces anuratus CK-J was investigated. The weight ratio of fermented organic matter containing chitin to zeolite was 11
Staged, adjusted the initial bacterial density to 10 ⁴ cfu / g of material,
The results of measuring the number of bacteria after culturing in an open system for 10 days are shown in Table 2.

[0019]

[Table 2]

From the results of this test, the mixture of chitin-containing fermented organic matter and zeolite may have any weight ratio from 99: 1 to 1:99, and particularly 2: 8 and 1: 9 were good. Example 3 In the initial research stage of biological control of soil diseases, a method of culturing an antagonistic microorganism and applying the bacterial cell to soil was often used, but the antagonistic microorganism was attacked by an indigenous microorganism and could be established for a long time. And most of these tests have failed. Here, Streptomyces anuratus CK-J (1 x Streptomyces anuratus CK-J 4 × 10 ⁹ per 1 g) grown in a mixture of chitin-containing fermented organic matter and zeolite (weight ratio 1: 1) (hereinafter “book”) When "invention material" was applied to the soil, it was examined whether the present microorganism can settle in the soil. -Test method Test soil Two types of soil were used: Kuroboku soil collected in Kurokawa Village, Niigata Prefecture and alluvial soil collected in Niitsu City, Niigata Prefecture. Test area A: Control area B: Inventive material 0.3% area C: Inventive material 1.0% area D: Inventive material 0.3% + chitin-containing fermented organic matter 0.7% area Experimental area contents A: 100 g soil, soil moisture It was adjusted to 55% of the maximum water capacity. Hereinafter, the soil moisture of B, C, and D was similarly treated. B: 0.3% of the material of the present invention was added to the soil. C: 1.0% of the material of the present invention was added to the soil. D: 0.3% of the material of the present invention and 0.7% of chitin-containing fermented organic matter were added to the soil. Incubation of soil The soil and materials were placed in an Erlenmeyer flask, adjusted to the water conditions of the field, covered with an aluminum foil, and kept at 25 ° C for 2, 5, 8 and 13 weeks. Bacterial count The number of actinomycetes and Streptomyces anuratus CK-J in soil was measured by the dilution plate method. Albumin agar medium was used for actinomycetes, and chitin medium was used for Streptomyces anuratus CK-J. Chitin was the only carbon source in the chitin medium, and since microorganisms other than this bacterium do not grow easily, it was used as the selective medium. -Results Changes in the number of Streptomyces anuratus CK-J and the total number of actinomycetes are shown in Figures 1 and 2. CK-J remained at a high bacterial density in all three experimental zones of materials B, C, and D, and the proportion of CK-J in all actinomycetes was high. The total number of actinomycetes of all three materials increased compared with the control group, and improvement of soil microbial flora was also expected. The inventive material 1.0% group shows a higher CK-J bacterial density and total actinomycete density than the inventive material 0.3% group, and the higher the applied amount of the material, the more advantageous it is for colonization and also for the growth of the actinomycetes group. It was advantageous. Moreover, rather than applying 1% of the material of the present invention alone, 0.3% of the material of the present invention and a fermented organic substance containing chitin
Since the CK-J bacterium density and the total actinomycete density are higher when 0.7% is applied, it is said that when the material of the present invention is applied to soil, the application of the chitin-containing fermented organic matter is more effective in controlling soil diseases. Judged

Example 4 The effect of controlling soil diseases of the material of the present invention was examined by a pot test. The target disease is tomato wilt disease, and the tomato wilt disease fungus ( Fu
sarium oxysporum f.sp. lycopersici race2) in soil 1
10 ⁵ propagules per gram of highly contaminated soil and 10 ⁴ propagul
Tomatoes (Momotaro; Sakata Seed Co., Ltd.) were cultivated on the weakly contaminated soil inoculated with es. For each contaminated soil, the material of the present invention was applied to the soil at 1%. For cultivation, a Wagner pot of 1/5000 are was used, and one tomato seedling with cotyledons developed was planted per pot, and 15 consecutive tests were conducted. After the cultivation was completed, the disease index was calculated by the following calculation formula, and the number of Fusarium bacteria in the soil was measured. The test results are shown in Table 3.

The formula for calculating the disease onset index is as follows. Disease index = {Σ (symptom index x number of individuals) / (total number of individuals 15 x
4)} × 100 disease symptoms Symptom index healthy 0 Cotyledon yellowing or drooping 1 Cotyledon withered deciduous leaf 2 Upper leaf withered deciduous leaf 3 Imminent or withered 4

[0023]

[Table 3]

From the results shown in Table 3, it was confirmed that the disease was reduced to about 2/3 by the soil application of the material of the present invention in the area where the disease was severely induced, and was completely suppressed in the area where the occurrence of the disease was relatively small. . It was also confirmed that the application of the material of the present invention reduced the density of Fusarium bacteria in the soil. Example 5 The control effect of strawberry chlorosis using the material of the present invention was examined in a field test. The early seedlings of Takako were planted on the contaminated soil in the vinyl house on June 10, 1990, and the rates of disease-causing strains on July 20, August 10, and August 30 were investigated. The test section was provided with a total of 3 sections, a control section, an application section of the present invention and a soil disinfection section. The application amount of the present invention material of the present invention material is about 10a
The soil disinfection area was 500 kg, and the soil was disinfected with 30 liters of chlorpicrin per 10a. 100 kg of chemical fertilizer (N: P: K = 10: 20: 10) was applied per 10 a in all three test plots. The number of fixed strains was 100 in each ward. The results are shown in Table 4.

[0025]

[Table 4]

As is clear from the test results in Table 4, it was confirmed that the application of the material of the present invention has an extremely high disease control effect comparable to soil disinfection.

[0027]

INDUSTRIAL APPLICABILITY According to the present invention, antagonistic actinomycetes capable of producing chitinase can be stably established in soil for a long period of time, and soil diseases can be effectively controlled.
Moreover, the present invention has a great advantage that it does not cause a problem of environmental pollution due to conventional chemical spraying.

[Brief description of drawings]

FIG. 1 is a diagram showing changes in the number of Streptomyces anuratus CK-J bacteria and the total number of actinomycetes in Kuroboku soil.

[Fig.2] Streptomyces anuratus in alluvial soil
It is a figure which shows the change of the number of CK-J bacteria, and the total number of actinomycetes.

Claims (3)

[Claims] 1. A soil disease controlling material comprising a fermented product obtained by fermenting a chitin or a chitin-containing organic matter and a plant organic matter, zeolite, and Streptomyces anuratus, which is an antagonistic actinomycete capable of producing chitinase. 2. A fermented product obtained by fermenting Streptomyces anuratus, which is an antagonistic actinomycete capable of producing chitinase, with a chitin or a chitin-containing organic matter and a plant organic matter and a zeolite are in a weight ratio of 99 :. A soil disease controlling material obtained by proliferating in a culture medium of 1 to 1:99. 3. A method for controlling soil diseases, which comprises applying the soil disease controlling material according to claim 1 or 2 to soil.