JP2023060853A - Fermented barley bran-containing soil improvement agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a fermented barley bran-containing soil improvement agent that solves the problem in which a barley bran alone has a low fertilizer component content, and no effective utilization method has been found other than improving soil physical properties.

SOLUTION: The present invention provides a soil improvement agent containing fermented barley bran. Further, as another aspect of the present invention, provided are a culture soil containing the soil improvement agent, a culture soil production method comprising a step of adding the soil improvement agent to the soil, a soil improvement kit comprising a fermented barley bran-containing soil improvement agent and a composition containing a bacterium of the genus Leptosphaeria, a strain deposited as receipt number NITE P-03000, and a composition containing the strain.

SELECTED DRAWING: Figure 7

COPYRIGHT: (C)2023,JPO&INPIT

Description

There is an application for the application of Article 30, Paragraph 2 of the Patent Act. Publisher name: The Japanese Society for Soil and Fertilizer Science Kanto Branch Niigata Conference Steering Committee Publication name: 2018 Japanese Society for Soil and Fertilizer Science Kanto Branch Conference Lecture Abstracts Publication date: December 1, 2018 (Heisei 30) 2. Meeting name: 2018 Kanto Branch Conference of the Japan Society of Soil Fertilizers Date: December 1, 2018 Poster name: Fertilizing properties of barley bran and its effect on soil microorganisms 3. Meeting name: 2018 Kanto Branch Conference of the Japan Society of Soil and Fertilizers Date: December 1, 2018 Poster name: Diversity of barley root fungi and their effects on barley growth

The present invention relates to soil conditioners, particularly soil conditioners containing fermented barley bran.

Barley is a globally important grain that is used as feed in addition to familiar foods such as miso, barley tea, beer, and barley shochu. Demand for barley is increasing with rising health consciousness and an increase in the world population, but the area under cultivation has not expanded remarkably. Therefore, it is important to efficiently produce barley in a limited cultivation area.

In addition, when barley is processed into products, "barley bran" (barley bran) is discharged as a by-product of manufacturing. The amount of this by-product discharged is 80,000 tons per year, and most of it is currently discarded or used only as fertilizer. In addition, barley bran alone has a low content of fertilizer components, and no effective use other than improving soil physical properties has been found.

Patent Literature 1 discloses an agricultural material in which beer lees are used as a mushroom culture bed and then mixed with plant culture soil.

JP 2000-139206

However, the technique of Patent Document 1 has a problem in that it takes too much time to use it as a plant culture soil because it uses beer lees as a mushroom culture bed. In addition, it was unclear whether the technique of Patent Document 1 could be applied to barley bran.

As a result of extensive research, the inventors of the present invention have found that fermented barley bran, which is obtained by fermenting barley bran, has a higher fertilizing effect than unfermented barley bran.

The present invention is a soil conditioner comprising fermented barley bran.

By using this soil conditioner, it is possible to produce soil that can promote the growth of plants.

Moreover, the soil conditioner may further contain a nitrogenous fertilizer. The nitrogenous fertilizer may be urea. The soil improver may further contain a fungus belonging to the genus Leptosphaeria. The fungus may infect the fermented barley bran. The bacterium may be the bacterium deposited under accession number NITE P-03000.

Further, as another aspect of the present invention, there is provided culture soil containing the above soil conditioner.

The soil conditioner may be added to the soil such that the total nitrogen derived from the fermented barley bran is 0.01 to 1.5 parts by mass per 100 parts by mass of soil.

Moreover, as another aspect of the present invention, there is provided a culture soil manufacturing method including the step of adding the above soil conditioner to soil.

The step of adding the soil conditioner to the soil is a step of adding the soil conditioner to the soil so that the total nitrogen derived from fermented barley bran is 0.01 to 1.5 parts by mass per 100 parts by mass of soil. may be

Further, as another aspect of the present invention, there is provided a soil improvement kit comprising a soil conditioner containing fermented barley bran and a composition containing Leptosphaeria spp. Also provided as another aspect of the invention is the strain deposited under Accession No. NITE P-03000. Another aspect of the invention provides compositions comprising the above strains.

FIG. 1 is a graph showing the nitrogen mineralization rate in soil for each of samples (2) to (6). FIG. 2 is a diagram showing the experimental method of Experiment 3 in the example. FIG. 3 is a graph showing the number of filamentous fungi counted over time in Experiment 3 of the example. FIG. 4 is a graph showing the number of bacteria counted over time in Experiment 3 of the example. FIG. 5 is a graph showing only control (1), unfermented barley bran (2) and fermented barley bran (4) from FIG. FIG. 6 is a graph showing only control (1), unfermented barley bran (2) and fermented barley bran (4) from FIG. FIG. 7 is a photograph showing the growth results of barley in Experiment 4 in the example. FIG. 8 is a graph showing the above-ground fresh weight, above-ground dry weight, root-dry weight and panicle number of barley in Experiment 4 in the example. FIG. 9 is a graph showing the fresh weight and dry weight of barley aerial part in Experiment 5 in the example. FIG. 10 is a graph showing the fresh weight and dry weight of barley ear in experiment 5 in the example. FIG. 11 is a graph showing the weight of the aboveground parts and roots of barley in Experiment 6 in the example. FIG. 12 is a graph showing the weight of the above-ground parts and roots of Japanese mustard spinach in Experiment 6 in the example. FIG. 13 is a graph showing the weight of the top part and root of sunny lettuce in Experiment 6 in the example.

Definitions For convenience, certain terms used in this application are collected here. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Unless the context clearly dictates otherwise, the singular forms "a,""an," and "the" include plural references.

Although the numerical ranges and parameters set forth herein are approximations, the numerical values set forth in the specific examples are described as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Also, as used herein, the term "about" generally means within 10%, 5%, 1% or 0.5% of a given value or range. Alternatively, the term "about" means within an acceptable standard error, as considered by one of ordinary skill in the art.

Embodiments of the present invention will be described below. The following embodiments are examples, and the scope of the present invention is not limited to those shown in the following embodiments. In addition, in order to avoid complication of repetition about the same content, the brief description is abbreviate|omitted.

Soil Conditioner Comprising Fermented Barley Bran In the present embodiment, a soil conditioner comprising fermented barley bran is provided. Adding soil conditioners to the soil can promote the growth of crops (eg, barley, Japanese mustard spinach, and sunny lettuce). In this embodiment, the barley may be, but is not limited to, two-rowed, four-rowed, six-rowed, bare or wild barley.

Barley bran is exfoliated products such as pericarp, seed coat and germ produced when barley is refined, and may contain endosperm during the polishing process. In some embodiments, it is preferred to use barley bran that has not been used in culture beds such as mushroom cultivation and/or has not been carbonized.

Fermented barley bran is obtained by fermenting barley bran. Barley bran may be sterilized by autoclaving. It can be produced from a mixing step of obtaining a mixture of fermentation starter and barley bran, a hydrating step of adding water to the mixture, and a fermentation step of fermenting the barley bran in the mixture. Fermentation starter and barley bran in a ratio of 1:0.5 to 100 (parts by weight), 1:1 to 80 (parts by weight), 1:5 to 50 (parts by weight) or 1:10 to 30 (parts by weight) part). In the hydration step, water is preferably added such that the barley bran has a moisture content of 30% to 60%, 35 to 55% or 40% to 50%. During the fermentation process, the moisture content of the barley bran is preferably maintained at 30% to 60%, 35 to 55% or 40% to 50%. In the fermentation process, the temperature of the barley bran is preferably maintained between 20°C and 35°C, or between 20°C and 30°C. In the fermentation process, fermentation may be carried out for 10 days to 60 days, and from 10 days, 15 days, 20 days, 25 days, 30 days, 35 days, 40 days, 45 days, 50 days, 55 days and 60 days. It may fall within the range between any two selected points. During the fermentation process, the barley bran may be agitated as appropriate (eg, daily, once every two days, and once every three days). The fermentation method may be natural fermentation or microbial fermentation, and the fermentation starter may be soil or a carrier (eg, medium and autoclaved soil) containing microorganisms (eg, yeast and filamentous fungi).

In one embodiment, the fermented barley bran may have a reduced carbon ratio (CN ratio) compared to the same lot of barley bran before fermentation. For example, the fermented barley bran may have a carbon ratio (CN ratio) lower than that of the same lot of barley bran before fermentation within the range of 1% to 50%. , 3%, 4%, 5%, 7%, 10%, 15%, 20%, 30%, 40% and 50%. . In another embodiment, the carbon fraction (CN ratio) of the fermented barley bran may be 20 or less, 18 or less, 15 or less, 13 or less, or 10 or less. Fermented barley bran may be sterilized by autoclaving. In some embodiments, the fermented barley bran has not been used in culture beds such as mushroom cultivation and/or has not been carbonized.

Fermented barley bran may be present in the soil conditioner in the range of 0.5% to 100% by weight, depending on the plant to be grown, 0.5%, 1%, 5%, 10% by weight. , 20% by mass, 30% by mass, 40% by mass, 50% by mass, 60% by mass, 70% by mass, 80% by mass, 90% by mass, 95% by mass, 98% by mass, 99% by mass, 99.9% by mass and 100% by mass It may be present in a range between any two points selected from % by mass.

Conventionally known additives such as surfactants, pH adjusters, solidifying agents, fats and oils, mineral materials, and plant fiber materials may be added to the soil conditioner as long as they do not impair the effects of the present invention.

Surfactants include anionic surfactants (e.g., soap, sulfated oils, polyoxyethylene alkyl ether sulfates, alkyl sulfates, alkylbenzenesulfonates, alkanesulfonates, α-olefinsulfonates, N - acylamino acid salts, dialkylsulfosuccinates, alkylnaphthalenesulfonates), cationic surfactants (e.g. alkyltrimethylammonium salts, alkylpyridinium salts), nonionic surfactants (e.g. polyoxyethylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene fatty acid esters, polyhydric alcohol fatty acid esters) and amphoteric surfactants (eg, betaine, sulfobetaine), etc., but not limited thereto.

Examples of pH adjusters include slaked lime, calcium carbonate, ammonium sulfate, phosphoric acid, and citric acid.

Examples of solidifying agents include sugar-based polymer compounds (e.g., carboxymethylcellulose, methylcellulose, chitosan), polyol-based polymer compounds (e.g., polyvinyl alcohol), acetic acid-based polymer compounds (e.g., polyvinyl acetate), and natural rubber. (e.g., gum arabic), polyvinylpyrrolidones, acrylic polymer compounds (e.g., acrylic acid (salt), 2-acrylamido-2-methylpropanesulfonic acid (salt), homopolymers or copolymers such as acrylamide ), natural polymer compounds (eg, xanthan gum, alginic acid), but not limited thereto.

Fats and oils include vegetable oils (eg, rapeseed oil, sesame oil, safflower oil, olive oil, almond oil, sunflower oil), fish oils (eg, whale oil, sardine oil, saury oil, herring oil).

Examples of mineral materials include vermiculite, perlite, and zeolite, but are not limited to these.

Examples of plant-based fiber materials include peat moss, coconut peat, coconut shell peat moss, bacchus, and bark, but are not limited to these.

The soil conditioner according to this embodiment may contain fertilizer or may be used together with fertilizer depending on the plant to be grown. Fertilizers include nitrogenous fertilizers (e.g., urea, ammonium sulfate, ammonium chloride, lime nitrogen), phosphate fertilizers (e.g., superphosphate, heavy superphosphate, molten phosphate, calcined phosphate), potassium Non-limiting examples include high-quality fertilizers (eg, potassium sulfate, potassium chloride, potassium bicarbonate) and compound fertilizers thereof.

The soil conditioner according to the present embodiment may contain fungi (filamentous fungi) belonging to the genus Leptosphaeria. In some embodiments, the fermented barley bran in the soil conditioner is infected with Leptosphaeria. Fungi of the genus Leptosphaeria infect fermented barley bran, for example, by inoculating autoclaved fermented barley bran and growing to 1 x 10^9 cfu/g (e.g., 2 to 4 weeks at 25°C). be able to. In another embodiment, the soil conditioner may be used with fungi of the genus Leptosphaeria (filamentous fungi). The Leptosphaeria genus may be used as a strain, or may be used as any composition containing the Leptosphaeria genus bacteria (eg, liquid agar growth medium, agar growth medium). In certain embodiments, the Leptosphaeria spp. is the strain deposited under Accession No. NITE P-03000.

The soil improver according to this embodiment may be in the form of powder, granules, pellets, or flakes. In some embodiments, the soil conditioner may consist solely of fermented barley bran. In another embodiment, the soil conditioner may consist solely of fermented barley bran and Leptosphaeria. In yet another embodiment, the soil conditioner has only fermented barley bran as grain-derived bran (including meal and bran).

Another embodiment provides a culture medium containing the above soil conditioner. The soil conditioner contains 0.01 to 1.5 parts by weight of total nitrogen derived from fermented barley bran per 100 parts by weight of soil, or 0.01, 0.02, 0.05, 0.2, 0.75 and 1.5 parts by weight. may be added to the soil within the range between any two points selected from

Kit In yet another embodiment, there is provided a soil improvement kit comprising the soil conditioner containing the fermented barley bran and a composition containing Leptosphaeria.

The kit may contain additives and/or fertilizers. Preferably, the fermented barley bran and the composition comprising the Leptosphaeria are each contained in a suitable container. The kit may include additional elements (eg, instructions including an expiration date).

Cultivation soil manufacturing method Another embodiment provides a culture soil manufacturing method including the step of adding the above soil conditioner to soil. In one embodiment, the step of adding the soil conditioner to the soil includes adding the soil conditioner in an amount of 0.01 to 1.5 parts by weight per 100 parts by weight of soil or 0.01 parts by weight, 0.02 parts by weight of total nitrogen from fermented barley bran. 0.05 parts by weight, 0.2 parts by weight, 0.75 parts by weight and 1.5 parts by weight.

Experiment 1 Fertilizer component evaluation Total nitrogen, total carbon, total potassium and total phosphorus contents in barley bran (fiber snow), rice bran and oil cake were measured. Total nitrogen content and total carbon content were measured by NC analyzer using pulverized samples (barley bran, rice bran and oil cake). Total potassium and total phosphorus were measured by ICP emission spectrometry. In the analysis by ICP emission spectrometry, a measurement sample was prepared by the following steps.
(1) Add 5 mL of concentrated nitric acid to 0.5 g of sample.
(2) Heat the solution of (1) at 120°C for 90 minutes.
(3) Quantify the solution of (2) to 50 mL with ultrapure water.
(4) Filter the solution of (3) through No.5 filter paper.
(5) Use the filtrate obtained in (4) as a measurement sample.
Table 1 shows the measurement results.

As shown in Table 1, barley bran has a lower total nitrogen (N), phosphorus (P) and potassium (K) content than rice bran and oil cake, and a carbon content (CN ratio) was high.

As described above, since barley bran has a low total nitrogen content, it was expected that the use of barley bran as fertilizer would cause nitrogen deficiency in plants, resulting in poor growth. Barley bran has a low total nitrogen content but a high CN ratio, suggesting the possibility of lowering the CN ratio by fermenting barley bran.

Experiment 2 Measurement of Inorganic Nitrogen Content in Soil The mineralization pattern of nitrogen in barley bran was investigated. Unfermented barley bran, fermented barley bran, unfermented rice bran, fermented rice bran and oil cake were used as samples. Fermented barley bran was produced by the following method. 1 g of soil was mixed with 20 g of barley bran, distilled water was added so that the moisture content of the barley bran was 45%, and the mixture was cultured at 25°C. Fermented barley bran was obtained by turning the barley once every two days and adding distilled water to keep the moisture content at 45% and fermenting for 30 days. Fermented rice bran was produced according to the method for producing fermented barley bran. Each sample was added to 400 g of soil and statically cultured at 25°C. As a control, 400 g of soil to which no sample was added was also used in the experiment. Table 2 shows the application rate for each sample (3 samples for each fertilizer plot).

5.0 g of control (1) and samples (2) to (6) soil were sampled. 50 mL of KCl was added to the sampled soil to obtain a filtrate. The amount of inorganic nitrogen was measured from the filtrate. The amount of nitrate nitrogen among inorganic nitrogen was measured by the alkali reduction/diazo dye method. Among the inorganic nitrogens, the amount of ammonia nitrogen was measured by the indophenol blue method. The measurement results are shown in FIG.

As shown in Fig. 1, barley bran had a lower mineralization rate than rice bran and oil cake. In both unfermented barley bran and unfermented rice bran, organicization of nitrogen occurs (the rate of nitrogen mineralization becomes negative), but unfermented barley bran turns to mineralization later than unfermented rice bran. . It was found that fermented barley bran is more useful than unfermented barley bran because fermented barley bran maintains a positive nitrogen mineralization rate and does not cause nitrogen to become organic.

Experiment 3 Soil Microbial Count The number of microorganisms in the soil used in Experiment 2 was counted. 0.5 g of the soil used in Experiment 2 was sampled. Sterilized water was added to the sampled soil and quantified to 5.0 mL. As shown in FIG. 2, 10-fold dilution was repeated with sterilized water, and 50 μL or 100 μL of the soil suspension was sampled at a specific dilution rate and inoculated onto an agar medium. Rose Bengal medium was used for fungi (100 µL of inoculum). R2A medium was used for bacteria (inoculum volume was 50 μL). The number of colonies of growing bacteria and filamentous fungi was counted and converted to the number of bacteria per 1 g of dry soil. The results are shown in Figures 3-6.

FIG. 3 shows filamentous fungal counts. In the fermented rice bran (5) and the fermented barley bran (4), the number of filamentous fungi increased remarkably, and the number of filamentous fungi increased rapidly from 0 to 7 days, and thereafter, the number of fungi was maintained from the 14th day. Control (1), unfermented rice bran (3), unfermented barley bran (2) and oil cake (6) did not show a significant increase in filamentous fungi.

FIG. 4 shows bacterial counts. The number of bacteria increased in unfermented barley bran (2) and unfermented rice bran (3). An increase in the number of bacteria was observed in the order of rice bran>barley bran>oil cake.

FIG. 5 shows the number of filamentous fungi only in control (1), unfermented barley bran (2) and fermented barley bran (4) from FIG. (2) and fermented barley bran (4) only show bacterial counts. As is clear from FIGS. 5 and 6, in the fermented barley bran (4), filamentous fungi increased first, and then bacteria increased. In unfermented barley bran (2), an increase in the number of bacteria was observed, but no increase in filamentous fungi was observed.

Experiment 4 Barley Cultivation Test The samples shown in Table 3 were added to the soil to cultivate barley.

Germinated barley (3 days) was planted in 400 g of soil fertilized with each fertilizer and grown for 28 days. FIG. 7 is a photograph showing the growth results. The grown barley was harvested, and the fresh weight of the aboveground part, the dry weight of the aboveground part, the root-dry weight and the number of panicles were examined. The results are shown in FIG.

As shown in FIG. 8, the fermented barley bran + urea had significantly higher aerial fresh weight, dry weight and panicle number values compared to the control. There was no significant difference in dry weight of above ground parts, dry weight of roots and panicle number compared with chemical fertilizers. Therefore, it was clarified that fermented barley bran and fermented barley bran + urea are useful as uses.

Experiment 5 Isolation test of barley root endophytic fungi Useful endophytic fungi colonizing barley roots were selected. Barley was collected in Yamanashi, Ibaraki, Gunma and Toyama prefectures. The harvested barley roots were cut with scissors. The cut roots were added to 70% ethanol, then soaked in 1% sodium hypochlorite solution and washed three times with sterile water. Some of the washed roots were cryopreserved at -80°C, and the rest of the roots were cultured on an antibiotic-containing agar medium to isolate filamentous fungi. A total of 25 filamentous fungi were isolated.

Barley seeds were germinated and infected with the isolated filamentous fungi, and their growth was observed. Barley seeds were stirred overnight in sterile water, then the seeds were soaked in a 1% calcium hypochlorite solution and the seeds were washed three times with sterile water. Seeds were sown on an agar medium and infected with the isolated filamentous fungi. Barley seeds were infected with Piriformospora indica (P. indica) as a positive control. After germination, barley was grown in a chemically fertilized soil (N: 2.4 mg/100g-soil) and in non-fertilized soil in a climate chamber. After 28 days of growth, fresh and dry weights of barley tops and ears were compared. As shown in Figures 9 and 10, filamentous fungus No. 23 exhibited significant growth-promoting resistance.

The barley root from which No. 23 filamentous fungus was isolated was pulverized with metal beads and subjected to DNA extraction. The ITS region of the extracted DNA was amplified by PCR and analyzed using a sequencer. As a result, the filamentous fungus No. 23 was found to belong to the genus Leptosphaeria.

No. 23 filamentous fungus was deposited at the National Institute of Technology and Evaluation Patent Microorganism Depositary Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) on July 5, 2019 (receipt date) under the receipt number NITE P. Deposited as -03000.

Experiment 6 Cultivation experiment using filamentous fungus No. 23
Barley, Japanese mustard spinach, and sunny lettuce were cultivated using barley bran cultured with filamentous fungus No. 23. After autoclave sterilization of barley bran and inoculation with filamentous fungus No. 23, it was grown at 25°C for 3 weeks to grow to 1 x 10^9 cfu/g. Barley, Japanese mustard spinach, and sunny lettuce were cultivated using barley bran infected with filamentous fungus No. 23. Cultivation conditions are shown in Table 4.

FIG. 11 shows cultivation results in barley. When barley bran cultured with filamentous fungus No. 23 was used, it was found that the weight of the aboveground parts and roots of barley increased remarkably.

FIG. 12 shows the cultivation results for Komatsuna. It was found that when barley bran cultured with No. 23 filamentous fungus was used, the length of the plant height and the weight of the top and root parts of Komatsuna remarkably increased.

FIG. 13 shows the cultivation results for sunny lettuce. It was found that when barley bran cultured with No. 23 filamentous fungus was used, the height of the sunny lettuce and the weight of the above-ground part increased remarkably.

The filamentous fungus of No. 23 did not cause disease in any of barley, Komatsuna and sunny lettuce, and could be grown safely.

Claims (13)

A soil conditioner comprising fermented barley bran. 2. The soil conditioner of claim 1, further comprising a nitrogenous fertilizer. The soil conditioner according to claim 2, wherein said nitrogenous fertilizer is urea. The soil conditioner according to any one of claims 1 to 3, further comprising a fungus belonging to the genus Leptosphaeria. 5. The soil conditioner according to claim 4, wherein said fungus infects said fermented barley bran. The soil conditioner according to claim 4 or 5, wherein the fungus is deposited under accession number NITE P-03000. Cultivation soil containing the soil conditioner according to any one of claims 1 to 6. 8. The soil conditioner of claim 7, wherein the soil conditioner is added to the soil such that the total nitrogen from the fermented barley bran is 0.01 to 1.5 parts by weight per 100 parts by weight of soil. potting soil. A method for producing culture soil, comprising the step of adding the soil conditioner according to any one of claims 1 to 6 to soil. The step of adding the soil conditioner to the soil is a step of adding the soil conditioner to the soil so that the total nitrogen derived from fermented barley bran is 0.01 to 1.5 parts by mass per 100 parts by mass of soil. The method for producing culture soil according to claim 9. a soil conditioner comprising fermented barley bran;
a composition comprising a bacterium belonging to the genus Leptosphaeria;
Soil amendment kit comprising: The strain deposited under accession number NITE P-03000. A composition comprising the strain of claim 12.

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