JPH11225747A - Microorganism degrading organic material, culturing material composition thereof, and prevention of fairy ring disease by using the culturing material composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new microorganism useful for degradation of organic materials such as composting of organic materials such as straw, bark and waste of stock raising, and degradation promotion of garbage. SOLUTION: This new microorganism is selected from Penicillium sp. CF-1 (FERM P-16479), Penicillium sp. FS-26 (FERM P-16480) and Penicillium sp. FS-29 (FERM P-16481), having cellulose-degrading activities, and Bacillus sp. LB-5 (FERM P-16485), Debaryomyces sp. LB-31 (FERM P-16484), Aspergillus sp. LF-3 (FERM P-16482) and Aspergillus sp. FS-35 (FERM P-16483), having lignin-degrading activities. The cultivating material composition is preferably prepared by including one or more kinds of the new microorganisms and a cultivating material such as vermiculite.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel microorganism and a culture material composition to which the novel microorganism is added, and is useful for composting organic matter such as rice straw, bark, livestock waste, and decomposing organic matter such as accelerating the decomposition of garbage. It is something. Further, the present invention relates to a method for controlling turf fairy ring disease using the culture material composition.

[0002]

2. Description of the Related Art Conventionally, calcareous materials such as lime nitrogen and magnesite lime are generally used as materials for promoting the maturation of organic substances. In recent years, many materials using microorganisms and enzymes have been distributed. However, when lime nitrogen or formic lime is added, the alkali is excessive due to the increase in pH accompanying the decomposition of organic matter. On the other hand, very few materials using microorganisms or enzymes clarify the type of microorganism added, and the mechanism of action is not clear. Under these circumstances, there is a need for more effective ones using microorganisms.

[0003] In a golf course, fairy grass disease occurs due to the accumulation of cut grass and other organic matter in the soil, and chemical pesticides are used to control this. However, basidiomycetes that have formed white soil easily die. And the appearance of turfgrass is greatly impaired. Ultimately, the turf needs to be renewed, and the economic loss is large. Therefore, a method for controlling fairy ring disease that does not rely on chemical pesticides is desired.

[0004] Fairy ring disease is a disease in which turfgrass wilts in a circular or arc shape, and occurs when a large amount of basidiomycetes inhabits thatch and soil. This pathogen is mainly a Basidiomycete fungus. This fairy ring disease is susceptible to damage on fairways with low moisture and fertilizer, turfs and sand greens with a developed thatch layer.

[0005]

The problems to be solved by the present invention are novel microorganisms which degrade cellulose and lignin which are difficult to decompose, woody materials such as sawdust and bark using these microorganisms, rice straw and wheat straw. Grass residues such as straws, peaches, bran, etc., pruned branches such as tea and fruit trees, crop residues such as vegetables, cut grass from golf courses, garbage excreted from homes, livestock excrement, food industry waste, etc. It is an object of the present invention to provide a culture material composition for accelerating the decomposition of turf, and an agent for controlling turf fairy ring disease containing the microorganism.

[0006]

That is, the present invention relates to Penicillium sp.
lium sp. FS-26 or Penicillium sp. FS-29.

Further, the present invention has a lignin decomposability.
Bacillus sp. LB-5.

Further, the present invention has lignin decomposability.
Debaryomyces sp. LB-31.

Further, the present invention has lignin decomposability.
Aspergillus sp. LF-3 or Aspergillus sp. FS-3
5

Further, the present invention is a culture material composition containing at least one of the above microorganisms and a culture material.

Further, the present invention is a culture material composition comprising the above microorganism, at least one of Bacillus sp. BS-1SMCP III having cellulolytic ability, and a culture material.

As the culture material, vermiculite,
Inorganic minerals such as zeolite, perlite, montmorillonite, ammonium fertilizers such as ammonium sulfate, ammonium chloride, ammonium nitrate, lime superphosphate, ammonium phosphate, potassium sulfate, potassium chloride, iron compounds such as ferrous sulfate and ferrous chloride, and diatomaceous Porous materials such as soil, calcined diatomaceous earth, coral sand, charcoal powder, and activated carbon; vegetable fertilizers such as rapeseed oil cake, rice bran oil cake, castor oil cake, and soybean oil cake, steamed bone meal, raw bone meal, deflocculated bone meal, and meat Examples include those selected from the group consisting of animal fertilizers such as bone meal, meat meal, fish meal, feather meal, steamed hair meal, crab, skin powder, and pupa, and mixtures thereof.

Further, the present invention is a culture material composition, wherein the microorganism is cultured in the culture material.

Further, the present invention relates to a method for controlling fairy fairy disease of turf using the culture material composition.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

The method for separating and selecting lignin- or cellulose-degrading microorganisms of the present invention and the method for producing a culture material composition containing these microorganisms will be described.

[Isolation of Cellulose-Decomposing Bacteria] Strains isolated from soil, compost, etc. collected from various places by a dilution plate method were subjected to CMC (carboxymethylcellulose) -containing plate medium (5 g of CMC, ₄ g of KH ₂ PO 4, Na ₂ HPO _4). 6 g, (NH ₄ ) ₂ SO ₄ 2 g,
_{MgSO 4 · 7H 2 O 200mg,} CaCl 2 1mg, Fe 2 (SO 4) · 7H 2 O, yeast extract _{1g, H 3 BO 3 10μg,} MnSO 4 10μg, ZnSO 4 70μ
_{g, CuSO 4 50μg, MoO 3} 10μg, agar 15 g, distilled water 1000
The strain was cultured at 10 ° C., 30 ° C., and 50 ° C., and a strain having a clear zone formed on the medium was selected. The selected strain was added to 50 mL of an inorganic salt liquid medium containing 0.5% CMC, cultured at 10 ° C., 30 ° C., and 50 ° C. for 1 week, and the supernatant obtained by centrifuging the culture was subjected to the following analysis. That is, the total amount of sugar in the obtained supernatant is quantified by the phenol sulfate method, and the amount of reducing sugar is quantified by the modified Somogi method. Was selected as At the same time, the rice straw was pulverized and urea was added to adjust the carbon content. Then, distilled water equivalent to the maximum water retention of the rice straw was added, and the mixture was sterilized in an autoclave at 121 ° C. for 30 minutes. The strain was added thereto, and the carbon ratio after incubation at the selected temperature for 4 weeks was measured. Table 1 shows the results.

[0018]

[Table 1]

[Isolation of Lignin-Decomposing Bacteria] From a soil, compost, etc. collected from various places, a strain isolated by a dilution plate method was converted into a ligninsulfonic acid-containing plate medium (ligninsulfonic acid Na 0.1 g, NH ₄ NO ₃ 1.0 g, K ₂ HPO ₄ 1.0g, MgSO ₄・ 7H ₂ O
Using 0.5 g, 1000 ml of distilled water, pH 6.8), the cells were cultured at 30 ° C., and primary selection was performed by selecting a Barbendam reaction-positive strain, a red reaction-positive strain in a guaiacol-supplemented medium, and selection by protocatechinic acid resolution.

The strain selected by the above method was transformed into a wood flour extraction liquid medium (wood flour 20 g, NH ₄ NO ₃ 1.0 g, K ₂ HPO ₄ 1.0 g, M
gSO ₄ · 7H ₂ O 0.5g, distilled water 1000 ml, was inoculated into pH 6.8) 30
Shaking culture was performed at 2 ° C. for 2 weeks, and the supernatant obtained by centrifuging the culture was subjected to the following analysis. That is, the total amount of sugar in the obtained supernatant was quantified by the phenol sulfate method, the amount of reducing sugar was modified by the Somogi method, the protein was determined by the Lowry method, and the ultraviolet absorbing substance was measured by absorbance measurement at 280 nm. Table 2 shows the results.

[0021]

[Table 2]

The origin of the novel microorganism used in the present invention and its mycological properties will be described.

1) The LB-5 strain is a novel microorganism having lignin degradability at a high temperature of about 50 ° C. isolated from bark compost. The characteristics of this strain are as shown in Table 3. It belongs to Bacillus from morphological and physiological tests,
cillus azotoformans, which differed in glucose, mannitol assimilation, and starch degradation. Therefore, this strain was identified as a new species. This strain was designated as Bacillus sp. LB-5. This strain was transferred to FERM P-16485 by the National Institute of Bioscience and Human Technology.
Has been deposited.

[0024]

[Table 3]

2) The LB-31 strain is a novel microorganism having a lignin degrading ability at a low temperature around 10 ° C. isolated from Hokkaido Sarobetsu peat. The characteristics of this strain are as shown in Tables 4 and 5, which are classified into spore-forming yeasts, and are characterized by vegetative cells, spore morphology and formation status, colony morphology and color, etc.
Although it is a related species of ryomyces castellii, it differs due to the assimilation of sugar and the like. Therefore, this strain was identified as a new species.
This strain was named Debaryomyces sp. LB-31. This strain was fed to the Institute of Biotechnology,
Deposited as RM P-16484.

[0026]

[Table 4]

[0027]

[Table 5]

3) The strain CF-1 is a novel microorganism having a cellulose decomposing ability at a low temperature of about 10 ° C., which is isolated from decayed wood. The characteristics of this strain are as shown in Tables 6 and 7,
Penicillium glabrum is a closely related species due to its morphological characteristics and the like, but a part of the assimilation of sugar is different. Therefore, this strain was identified as a new species. This strain was designated as Penicillium sp. CF-1. This strain has been deposited with the National Institute of Bioscience and Biotechnology at the National Institute of Advanced Industrial Science and Technology as FERM P-16479.

[0029]

[Table 6]

[0030]

[Table 7]

4) The LF-3 strain was isolated from soil at 30 ° C.
It is a new microorganism that has lignin degradability in the middle temperature range before and after. The characteristics of this strain are as shown in Tables 8 and 9. Aspergillus nidulans is a closely related species based on the characteristics of the morphology, etc., but some of the assimilation of sugar is different. Therefore, this strain is recognized as equivalent to a new species. did. Then, this strain is called Aspergillu
ssp.LF-3. This strain has been deposited with the National Institute of Advanced Industrial Science and Technology as a FERM P-16482.

[0032]

[Table 8]

[0033]

[Table 9]

5) The FS-26 strain was isolated from peat.
It is a novel microorganism that has the ability to degrade cellulose in the low to medium temperature range of ℃ to 30 ℃. The characteristics of this strain are as shown in Tables 10 and 11, and from the characteristics of morphology and the like, it was determined to be a closely related species and Penicillium funicul
osum, but some of the assimilation properties of the sugars were different, so this strain was identified as equivalent to a new species. This strain was named Penicillium sp. FS-26. This strain has been deposited as FERM P-16480 with the National Institute of Bioscience and Human Technology.

[0035]

[Table 10]

[0036]

[Table 11]

6) The FS-29 strain is a novel microorganism having cellulolytic ability at a low to medium temperature range of 10 ° C. to 30 ° C. isolated from peat in Bibai City, Hokkaido. The characteristics of this strain are shown in Tables 12 and 1.
As shown in Fig. 3, Penicill is considered to be a closely related species due to its morphological characteristics.
ium glabrum, but with some differences in the assimilation of sugars. Therefore, this strain was identified as a new species equivalent. This strain was named Penicillium sp. FS-29. This strain was sent to FERM P-
Deposited as 16481.

[0038]

[Table 12]

[0039]

[Table 13]

7) FS-35 is a novel microorganism having a lignin degrading ability at a medium temperature range of about 30 ° C. isolated from Hokkaido Sarobetsu peat. The characteristics of this strain are as shown in Tables 14 and 15. Aspergillus te
rreus, but some of the sugar assimilation properties were different. Therefore, this strain was identified as a new species. This strain was named Aspergillus sp. FS-35. This strain was transferred to FERM P-16483 by the National Institute of Biotechnology and Industrial Technology.
Has been deposited.

[0041]

[Table 14]

[0042]

[Table 15]

8) Bacillus sp. BS-1SMCP III is disclosed in
As described in JP 91869, chloramphenicol-resistant microorganisms obtained by treating a strain isolated from rhizosphere soil with nitrosoguanidine are microorganisms having a cellulose-degrading ability at a medium temperature range of about 30 ° C. This strain has been deposited as FERM P-12516 with the National Institute of Biotechnology and Industrial Technology. The mycological properties are as follows.

[0044]

[Table 16]

9) The novel agent for controlling turf fairy ring disease of the present invention is a novel microorganism CF-1, CF-3, FS-26, F
S-29, FS-35, LB-5, LB-31 and BS-1SM
It is prepared by culturing at least one or more of CP III with a culture material.

Although this controlling agent can be used as it is,
Using a suitable solid carrier, liquid carrier, emulsifying dispersant, etc.,
It can be used in any form such as granules, powders, tablets, emulsions and wettable powders. At this time, the used amount of the culture of the microorganism is 0.1 to 50% based on the carrier. The control agent can be used together with an inorganic fertilizer, an organic fertilizer, a herbicide and the like.

[0047] The amount of the turf of golf courses and the like of the control agent, turf grass 1m ² per 1~1000g is preferable.

[0048]

EXAMPLES Hereinafter, cultures produced by using selected microorganisms will be described in more detail with reference to Examples, but the scope of the present invention is not limited to these Examples.

[Example 1] Novel microorganism C related to the present invention
F-1, CF-3, FS-26, FS-29, FS-3
5, LB-5, LB-31 and BS-1SMCP III were each added to a PD liquid medium (potato extract 20 mL, glucose 20 g, distilled water 100
(0 mL) at 30 ° C. for 7 days. After adding 5 L of mixed culture solution obtained by mixing 625 ml of this culture solution to 25 L of tap water, 100 kg of culture material (51 kg of rice bran, 10 kg of steamed bone meal,
Vermiculite 28 kg, charcoal 10 kg, ferrous sulfate 1 kg).

This culture material was deposited to a thickness of 10 cm, and cut back as appropriate so that the fermentation temperature did not exceed 40 ° C., and the added microorganism was grown. After that, the culture material is
It was air-dried until it became.

[Test Example] A test example using the test product manufactured in Example 1 is shown below.

[Test Example 1] Decomposition test of rice straw compost [Test plot]

[Test method] Straw 12kg in a 200l plastic container
And a composting test was conducted in the test plot. The test is reverted 16 days and 45 days after the test starts.

[Test Results] As shown in FIG. 1, the carbon decomposition rate of rice straw was large in the section where the test sample was added from the initial stage of deposition, and in particular, in the test sample + urea section, it was 30 after 36 days. % And over 50% on day 72. The test sample alone had the next highest decomposition rate.

[Test Example 2] Decomposition test in paddy field [Test plot]

[Test Method] Material treatment on October 12, November 16, December 22,
April 12 of the following year Collecting rice straw [Test results] As shown in Fig. 2, degradation of rice straw was high in the test sample addition zone from the initial stage of material application, and this tendency continued thereafter, especially in the low temperature season from December to April. The decomposition was excellent.

Test Example 3 Mushroom Waste Oga Decomposition Test [Test Method] Waste ogre (residue for cultivation of fungi and mushrooms such as enokitake mushrooms), cow manure and test products were mixed and packed in a mesh bag at the ratio shown in Table 1, and the sheet was protected from rain. For field deposition.

May 30 Deposition started, July 18 Interim sampling, August 29 Final sampling [Test plot]

[Test Results] The temperature of the product rapidly increased from the day after the deposition, and was maintained at 50 ° C. or higher for almost 10 days. After that, it was higher than the outside temperature at around 30 ° C. In the test plot (addition of test product), the product temperature remained high throughout the entire period, and the temperature difference in the initial stage of deposition was particularly large, and the decomposition promoting effect was high. (Fig. 3) As for the chemical properties, carbon has been reduced by about 13% in the treated area.
Regarding the microbial properties, the number of filamentous fungi derived from the test products was very large. (Table 17) From the above, the addition of the test product was effective for the early composting of waste sawdust.

[0060]

[Table 17]

[Test Example 4] Rice straw composting test [Test plot]

[Test Method] The above treatment was carried out using rice straw for 7.5 ha of paddy field in one ward and piled up.

Material treatment on November 15, December 24, April 25 of the next year Inawara collection [Results of test] The carbon ratio was 1 month after deposition, compared with 54.4 in the untreated section and the test article + urea section. Then it was 19.3, about 1/3 of the figure. Even after 5 months, the degree of decomposition did not change, and the carbon ratio decreased in the order of test article + urea section <test article section <urea section <untreated section (FIG. 4).

As a result of applying the compost subjected to each treatment to the soil and cultivating oats, no obstacle was observed except that the growth was suppressed slightly in the untreated plot. The growth of oats in the test product + urea group was significantly improved (FIG. 5).

[Test Example 5] Fairy ring control test [Test method] A test product was sprayed at a rate of 500 g / m ² around a golf course green at a place where fairy ring disease occurred. It was sprayed to 200 g / m ² .

[Test Results] At the place where the test product was sprayed,
The occurrence of fairy ring was suppressed, and the lawn grass grew normally (FIGS. 6 and 7).

[Example 2] Novel microorganism C related to the present invention
F-1, FS-35, LB-31, and BS-1 SMCP III were shaken at 30 ° C. for 7 days using PD liquid medium (potato extract 20 mL, glucose 20 g, distilled water 1000 mL). 5 L of a mixed culture obtained by mixing 1250 ml of this culture with an equal volume
After adding to L, 100 kg of culture material (51 kg of rice bran, steamed bone meal)
10kg, vermiculite 28kg, charcoal 10kg, ferrous sulfate 1k
g).

This culture material was deposited to a thickness of 10 cm, and cut back as appropriate so that the fermentation temperature did not exceed 40 ° C., and the added microorganism was grown. After that, the culture material is
It was air-dried until it became.

[Test Example] A test example using the test product manufactured in Example 2 is shown below.

[Test Example 1] Decomposition test in paddy field [Test plot]

[Test Method] Material treatment on October 30 and collection of rice straw on April 10 of the following year [Test results] As shown in Table 18, the carbon content indicating degradation of rice straw was low in the test plot, and thus C / N was low. It was the lowest.

[0072]

[Table 18]

[Test Example 2] Decomposition test of cut grass [Test plot]

[Test method] 2 kg of cut turf and 20 g of material were mixed in a 1 / 2000a Wagner pot, water was added thereto and the turf was deposited for 56 days, and nitrogen, carbon and microbial properties of the turf were measured.

[Test Results] As shown in Table 19, the carbon content indicating the decomposition of cut grass was low in the test plot, and the number of microorganisms exhibiting the decomposition activity was very large in the test plot.

[0076]

[Table 19]

[0077]

According to the present invention, woody materials such as sawdust and bark, straws such as rice straw and wheat straw, Japanese peach,
It is used to remove hard-to-decompose organic matter such as grain residues such as bran, pruned branches such as tea and fruit trees, harvest residues such as vegetables, cut grass from golf courses, raw garbage excreted from homes, livestock excrement, and food industry waste. Early decomposition is possible, and high quality compost can be obtained.
Further, the present invention can control fairy ring disease.

[Brief description of the drawings]

FIG. 1 is a view showing the carbon decomposition rate of deposited rice straw.

FIG. 2 is a view showing the carbon decomposition rate of rice straw during the test period.

FIG. 3 is a diagram showing a change in product temperature accompanying the accumulation of waste sawdust.

FIG. 4 is a view showing a carbon ratio of deposited rice straw.

FIG. 5 is a view showing the results of oat cultivation test using deposited rice straw.

FIG. 6 is a photograph showing turf withered due to fairy ring disease.

FIG. 7 is a photograph showing suppression of fairy ring disease by application of a test article.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI C12N 1/16 C12N 1/16 GD // C05F 11/08 C05F 11/08 (C12N 1/20 C12R 1:07) (C12N 1/14 C12R 1:80) (C12N 1/14 C12R 1:66) (C12N 1/16 C12R 1: 645) (72) Takanori Akazawa 5-551 Namiki, Tsuchiura-shi, Ibaraki Pref. Chika Katakura Tsukuba Co., Ltd. Within the Research Institute

Claims (10)

[Claims] 1. A microorganism selected from Penicillium sp. CF-1, which has a cellulose decomposing ability, Penicillium sp. FS-26 or Penicillium sp. FS-29. 2. Bacillus sp. LB-5 having lignin decomposability. 3. Debaryomyces sp. LB-31 having lignin degradability. 4. Aspergillus sp. LF-3 or Aspergillus sp. FS-35 having lignin decomposability. 5. A culture material composition comprising at least one of the microorganisms according to claim 1 and a culture material. 6. Bacillus sp. BS-1SMCP I and at least one of the microorganisms according to claims 1 to 4.
A culture material composition comprising II and a culture material. 7. The culture material is an inorganic mineral such as vermiculite, zeolite, perlite or montmorillonite; a chemical fertilizer such as ammonium sulfate, ammonium chloride, ammonium nitrate, lime superphosphate, ammonium phosphate, potassium sulfate or potassium chloride; ferrous sulfate. , Iron compounds such as ferrous chloride, diatomaceous earth, calcined diatomaceous earth, coral sand, charcoal powder, porous material such as activated carbon, rapeseed oil residue, rice bran oil residue, castor oil residue, plant fertilizer such as soybean oil residue, Steamed bone meal, raw bone meal, deglutinated bone meal, meat-and-bone meal,
7. A material selected from the group consisting of animal material fertilizers such as meat meal, fish meal, feather meal, steamed flour, crab, husk, pupa and the like, and a mixture thereof. Culture material composition. 8. Mineral minerals such as vermiculite, zeolite, perlite and montmorillonite; chemical fertilizers such as ammonium sulfate, ammonium chloride, ammonium nitrate, lime superphosphate, ammonium phosphate, potassium sulfate and potassium chloride; ferrous sulfate; Iron compounds such as iron, diatomaceous earth, calcined diatomaceous earth, coral sand, charcoal powder, porous materials such as activated carbon, rapeseed oil residue, rice bran oil residue, castor oil residue, plant fertilizers such as soybean oil residue,
A culture selected from the group consisting of animal and animal fertilizers such as steamed bone meal, raw bone meal, deglutinated bone meal, meat-and-bone meal, meat meal, fish cake, feather meal, steamed hair meal, crab, skin powder, and pupa meal, and mixtures thereof. A culture material composition obtained by culturing at least one of the microorganisms according to claims 1 to 4 in the material. 9. Mineral minerals such as vermiculite, zeolite, perlite and montmorillonite; chemical fertilizers such as ammonium sulfate, ammonium chloride, ammonium nitrate, lime superphosphate, ammonium phosphate, potassium sulfate and potassium chloride; ferrous sulfate; Iron compounds such as iron, diatomaceous earth, calcined diatomaceous earth, coral sand, charcoal powder, porous materials such as activated carbon, rapeseed oil residue, rice bran oil residue, castor oil residue, plant fertilizers such as soybean oil residue,
A culture selected from the group consisting of animal and animal fertilizers such as steamed bone meal, raw bone meal, deglutinated bone meal, meat-and-bone meal, meat meal, fish cake, feather meal, steamed hair meal, crab, skin powder, and pupa meal, and mixtures thereof. Bacillus sp. And at least one of the microorganisms according to claims 1 to 4 in the material.
p.) A culture material composition obtained by culturing BS-1SMCP III. 10. A method for controlling turf fairy ring disease, comprising using the culture material composition according to claim 5.