Agricultural compositions, seedling products, and methods for cultivating plants

Abstract

The objective is to provide agricultural compositions, seedling products, and methods for cultivating plants that facilitate plant growth. [Solution] The agricultural composition contains microorganisms of the genus Neobacillus. The seedling product comprises a growing medium and seedlings grown in the growing medium. At least one of the seedlings and the growing medium is treated with the agricultural composition. The method for cultivating plants comprises the step of treating the plants with the agricultural composition.

Description

Technical Field

[0001] The present invention relates to an agricultural composition, a nursery stock product, and a method for cultivating plants.

Background Art

[0002] Patent Document 1 discloses a microbial strain of the species Arthrobacter oxydans of the genus Arthrobacter, which has the properties of promoting the growth of Capsicum plants and suppressing the occurrence of diseases, and also has the property of preventing the yield reduction due to continuous cropping of soybean plants, with the accession number FERM P-22038.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] By using an agricultural composition containing the microbial strain disclosed in Patent Document 1, an effect of promoting the growth of specific plants can be obtained, but there is still room for further improvement in such an agricultural composition containing such a microorganism.

[0005] The present invention has been made in view of the above problems, and an object thereof is to provide an agricultural composition, a nursery stock product, and a method for cultivating plants that are easy to promote the growth of plants.

Means for Solving the Problems

[0006] According to a first aspect of the present invention, the agricultural composition contains a microorganism of the genus Neobacillus.

[0007] According to a second aspect of the present invention, the seedling product comprises a growing medium and a seedling cultivated in the growing medium. At least one of the seedling and the growing medium is treated with the agricultural composition described in the first aspect.

[0008] According to a third aspect of the present invention, a method for cultivating plants comprises the step of treating the plants with the agricultural composition described in the first aspect. [Effects of the Invention]

[0009] According to the present invention, it is possible to provide agricultural compositions, seedling products, and methods for cultivating plants that facilitate plant growth. [Brief explanation of the drawing]

[0010] [Figure 1] This figure illustrates the taxonomic position of the microorganisms contained in the agricultural composition according to this embodiment. [Figure 2] This figure shows the results of the first test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Figure 3] This figure shows the results of a second test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Figure 4] This figure shows the results of the third test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Figure 5] This figure shows the results of the fourth test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Figure 6] This figure shows the results of the fifth test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Figure 7] This figure shows the results of the sixth test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Figure 8] This figure shows the results of the seventh test concerning the microorganisms contained in the agricultural composition according to this embodiment. [Modes for carrying out the invention]

[0011] <First Embodiment: Agricultural Composition> The following describes an agricultural composition according to the first embodiment of the present invention. First, the inventor will briefly explain the process by which he came to complete the agricultural composition according to this embodiment.

[0012] Traditionally, compost, soil conditioners, and biopesticides utilizing microorganisms have been commercialized. For example, bacteria of the Bacillus family (formerly known as Bacillus) are common bacteria found in soil. Examples of Bacillus species used in agricultural compositions include Bacillus pumilus, Bacillus amyloliquefaciens, and Bacillus oryzicola.

[0013] However, conventional agricultural compositions containing microorganisms still have room for improvement. For example, while some microorganisms contained in agricultural compositions are known to promote the growth of specific plants, there is a problem in that the types of plants for which this growth-promoting effect is observed are limited. Furthermore, with conventional agricultural compositions containing microorganisms, the microorganisms applied to the soil do not settle in the soil, and therefore cannot fully exert their growth-promoting effect on plants, or even if they do exert a growth-promoting effect, the effect does not last long enough. In addition, because there are a great many types of microorganisms, there is a problem in that there are undiscovered microorganisms that are useful as agricultural compositions, or there are microorganisms that are already known but whose usefulness as agricultural compositions has not been verified.

[0014] In view of the above-mentioned problems, the inventors have discovered a microorganism capable of solving the above-mentioned problems and have verified the usefulness of the microorganism as an agricultural composition, thereby completing the agricultural composition according to this embodiment.

[0015] The agricultural composition according to this embodiment contains a microorganism belonging to the genus Neobacillus. More specifically, the microorganism contained in the agricultural composition according to this embodiment is of the species Neobacillus cucumis. Even more specifically, the microorganism contained in the agricultural composition according to this embodiment is strain B20 of the species Neobacillus cucumis. Hereinafter, unless otherwise specified, the microorganism contained in the agricultural composition according to this embodiment will be referred to as "strain B20". The agricultural composition according to this embodiment has the advantage of being likely to promote plant growth by comprising the above-mentioned microorganism.

[0016] Strain B20 was deposited on March 3, 2025, at the Patent Microorganisms Depositary Center (NPMD) of the National Institute of Technology and Evaluation (NITE) (Room 122, 2-5-8 Kazusa Kamashima, Kisarazu City, Chiba Prefecture) under the accession number: NITE P-04312.

[0017] Strain B20 can be obtained, for example, from commercially available horticultural soil, from a research institution, or by screening from nature such as soil. Also, strain B20 can be propagated, for example, by using PYAc medium as an agar medium or Y medium as a liquid medium, setting the pH before sterilization of the medium to 6.47, performing sterilization at a sterilization temperature of 121°C for 20 minutes, and performing shaking culture, which is aerobic culture, at a culture temperature of 30°C overnight. PYAc medium can be prepared by dissolving sodium acetate (2 g / L), yeast extract (2 g / L), and agar (1.5 g / L) in 1 L of Mineral Basal Medium (see Int. J. Syst. Bacteriol. 27(4), 346-348 (1977)). Also, Y medium can be prepared using yeast extract (3 g / L). For the composition of Mineral Basal Medium, refer to Table 5 described later. Also, for the composition of the Metal-EDTA solution contained in Mineral Basal Medium, refer to Table 6 described later. Note that the above culture method is an example and is not intended to limit the culture method of strain B20.

[0018] Figure 1 shows the results of identifying the microorganism (strain B20) by 16S rRNA amplicon sequencing. In Figure 1, the genus Neobacillus is represented by the old name, the genus Bacillus. As shown in Figure 1, the microorganism contained in the agricultural composition according to this embodiment (refer to "Bacillus sp. B20" enclosed by the solid-line rectangle in Figure 1) has no closely related species in the database and may be a new species. The species most closely related to the above microorganism was Neobacillus cucumis (formerly Batillus cucumis). Therefore, the microorganism contained in the agricultural composition according to this embodiment is also relatively preferable for Neobacillus cucumis.

[0019] Hereinafter, the characteristics of the agricultural composition according to this embodiment will be listed.

[0020] As shown in the results of the first test described below, the microorganism (here, strain B20) contained in the agricultural composition according to this embodiment has the property of growing in response to the root exudate of alfalfa. That is, it is considered that the microorganism contained in the agricultural composition according to this embodiment has the property of growing in response to the exudate of plant roots, not limited to the roots of alfalfa.

[0021] Also, as shown in the results of the second test described below, the microorganism (here, strain B20) contained in the agricultural composition according to this embodiment has the property of exerting a growth-promoting effect on Komatsuna. That is, it is considered that the microorganism contained in the agricultural composition according to this embodiment has the property of exerting a growth-promoting effect on plants of the genus Brassica, plants of the family Brassicaceae, and plants of the order Brassicales.

[0022] Furthermore, as shown in the results of the third test described later, the microorganism contained in the agricultural composition according to this embodiment (in this case, strain B20) has the property of exhibiting a growth-promoting effect on alfalfa. In other words, it is considered that the microorganism contained in the agricultural composition according to this embodiment also has the property of exhibiting a growth-promoting effect on plants of the genus Medicago, plants of the family Fabaceae, and plants of the order Fabales.

[0023] Furthermore, as shown in the results of the fourth test described later, the microorganism contained in the agricultural composition according to this embodiment (in this case, strain B20) has the property of exhibiting a growth-promoting effect on tomatoes. In other words, it is considered that the microorganism contained in the agricultural composition according to this embodiment also has the property of exhibiting a growth-promoting effect on plants of the Solanum genus, plants of the Solanaceae family, and plants of the Solanales order.

[0024] Thus, the microorganism contained in the agricultural composition according to this embodiment (here, strain B20) has the property of exhibiting a growth-promoting effect on at least one of the plants of the Brassicaceae family (here, komatsuna) and the Solanaceae family (here, tomato).

[0025] Furthermore, as shown in the results of the fifth test described later, the microorganism contained in the agricultural composition according to this embodiment (here, strain B20) has a turbidity of 0.05 or higher (OD 600 ) possesses properties that particularly promote plant growth. As will be described in detail later, when the microorganisms contained in the agricultural composition according to this embodiment have a turbidity of 0.05 or higher, preferably 0.1 or higher, and more preferably 0.5 or higher, a remarkable plant growth promoting effect can be expected.

[0026] Furthermore, as shown in the results of the sixth test described later, the microorganism contained in the agricultural composition according to this embodiment (in this case, strain B20) has the property of producing auxin.

[0027] Furthermore, as shown in the results of the seventh test described later, the microorganism contained in the agricultural composition according to this embodiment (in this case, strain B20) has the property of exhibiting a disease-suppressing effect against the bacterial wilt fungus (Ralstonia solanacearum). In other words, it is considered that the microorganism contained in the agricultural composition according to this embodiment has the property of exhibiting a disease-suppressing effect against plant pathogenic fungi.

[0028] As described above, the microorganism according to this embodiment (here, strain B20) not only exhibits a growth-promoting effect on a single type of plant, but also has the property of exhibiting a growth-promoting effect on multiple types of plants. In particular, strain B20 has the property of exhibiting a growth-promoting effect on any of the three types of plants: alfalfa, a plant of the genus Medicago in the family Fabaceae of the order Fabales; komatsuna, a plant of the genus Brassica in the family Brassicaceae of the order Brassicales; and tomato, a plant of the genus Solanum in the family Solanaceae of the order Solanales. In other words, strain B20 is considered to have the property of exhibiting a growth-promoting effect on any of the plants of the genus Medicago, Brassica, and Solanum. Furthermore, strain B20 is considered to have the property of exhibiting a growth-promoting effect on any of the plants of the family Fabaceae, Brassicaceae, and Solanaceae. Moreover, strain B20 is considered to have the property of exhibiting a growth-promoting effect on any of the plants of the order Fabales, Brassicales, and Solanales.

[0029] The agricultural composition according to this embodiment may contain at least a microorganism of the genus Neobacillus, particularly a microorganism of the species Neobacillus cucumis, and moreover, a microorganism of strain B20, as an active ingredient, but its form is not particularly limited. The microorganism contained in the agricultural composition may be in a dry state or a frozen state. Furthermore, the agricultural composition according to this embodiment may contain only the above-mentioned microorganism, but may further contain a carrier (e.g., water, buffer solution, culture medium, glycerol solution, or porous material), various additives (e.g., pH adjusters, humectants, preservatives, excipients, emulsifiers, or fungicides), and other components useful for plants (e.g., insecticides, herbicides, growth promoters, fertilizers, or antibiotics). Furthermore, the agricultural composition according to this embodiment may be, for example, a growing medium impregnated with the above-mentioned microorganism.

[0030] <Second Embodiment: Seedling Products> The following describes a seedling product according to a second embodiment of the present invention. The seedling product according to this embodiment comprises a growing medium and a seedling cultivated in the growing medium. Furthermore, in the seedling product according to this embodiment, at least one of the seedling and the growing medium is treated with the agricultural composition according to this embodiment.

[0031] In seedling products, at least one of the seedlings and the growing medium is treated with the above-mentioned agricultural composition, so that the surface of the seedling, the interior of the seedling, and at least a portion of the growing medium (hereinafter also referred to as the "treated area") contain a large amount of microorganisms derived from the above-mentioned agricultural composition (in this case, strain B20). For example, compared to commercially available seedling products of the same type that are not treated with the above-mentioned agricultural composition, the treated area of ​​the seedling product contains more than twice the amount (preferably more than five times the amount) of strain B20.

[0032] In seedling products, examples of plants used as seedlings include plants of the Brassicales order, plants of the Fabaceae order, or plants of the Solanales order. More preferably, examples of plants used as seedlings include plants of the Brassicaceae family, plants of the Fabaceae family, or plants of the Solanaceae family. More preferably, examples of plants used as seedlings include plants of the Brassica genus, plants of the Medicago genus, or plants of the Solanum genus. Even more preferably, examples of plants used as seedlings include Japanese mustard spinach, alfalfa, or tomato.

[0033] <Third Embodiment: Method for Cultivating Plants> The following describes a method for cultivating plants according to a third embodiment of the present invention. The method for cultivating plants according to this embodiment includes a step of treating plants with the agricultural composition according to this embodiment. Examples of plants cultivated by this method include the seedling plants described above.

[0034] Methods of treating plants with agricultural compositions include, for example, immersing plant seeds in a liquid agricultural composition, applying a liquid agricultural composition to the surface of a plant, pre-applying an agricultural composition to the soil in which the plant is cultivated, or spraying an agricultural composition onto a field where the plant is being cultivated. In addition, when cultivating plants by grafting, a method of using a rootstock plant treated with an agricultural composition as the rootstock for the plant to be cultivated (scion) is also possible.

[0035] In plant cultivation methods, it is preferable to treat plants with the agricultural composition as early as possible in the plant's life cycle, from the viewpoint of enhancing the disease suppression effect (i.e., the disease suppression effect of plant pathogenic fungi). For this reason, a preferred method of treating plants with the agricultural composition is to immerse the plant seeds in a liquid agricultural composition. The immersion time is, for example, 30 minutes to 4 hours. Note that the immersion time mentioned here is just an example and is not intended to limit the immersion time. (Examples)

[0036] The following describes specific embodiments of the present invention. However, the present invention is not limited to the following embodiments.

[0037] <First Exam> The following describes the contents and results of the first test concerning strain B20. The first test was to determine whether the microorganisms according to this embodiment increased in response to root exudate. Specifically, in the first test, 200 mg of soil containing strain B20 was prepared, and 80 μl of alfalfa root exudate was applied to the soil. After 24 hours, microbial DNA was extracted from the soil, and the relative abundance of strain B20 at the time of root exudate application and the relative abundance of strain B20 24 hours after root exudate application were compared by quantitative RT-PCR (Reverse Transcription-PCR).

[0038] Figure 2 shows the results of the first test. In Figure 2, "REMS activity" represents the proportion of B20 strains in the soil to which root extract (RE) was applied. As shown in Figure 2, the proportion of B20 strains 24 hours after application of root extract increased to approximately 3.28 times the proportion of B20 strains at the time of application of root extract (0h elapsed time).

[0039] Thus, the first test revealed that strain B20 has the property of amplifying in response to the exudate of plant roots (in this case, alfalfa roots). Therefore, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) have the advantage of being able to promote the growth of beneficial bacteria (in this case, strain B20) near plant roots, making it easier to create an environment in which the effects of beneficial bacteria can be sustained compared to agricultural compositions containing other bacteria.

[0040] <Second Exam> Next, we will explain the contents and results of the second trial concerning strain B20. The second trial was conducted to determine whether strain B20 has a growth-promoting effect on komatsuna, a plant belonging to the Brassicaceae family and Brassica genus of the Brassicales order. Specifically, in the second trial, 10 test plots were prepared in which komatsuna seeds were sown in commercially available vegetable soil (H-150 manufactured by Yanmar Agri Co., Ltd.), and 10 control plots were prepared. In each test plot, a bacterial suspension of strain B20 (turbidity (OD)) was added to the vegetable soil. 600 2 ml of (0.5) was applied. In contrast, no suspension of strain B20 was applied to the vegetable growing medium in each control group. After preparation of each test group and control group, komatsuna seedlings were grown for two weeks. Subsequently, the dry weight of the underground and above-ground parts of the grown komatsuna seedlings was measured in each test group and control group.

[0041] Figure 3 shows the results of the second experiment. The upper graph in Figure 3 shows the measurement results of the dry matter weight of the above-ground part of the komatsuna seedlings, and the lower graph in Figure 3 shows the measurement results of the dry matter weight of the underground part of the komatsuna seedlings. In Figure 3, "Control" represents the measurement results of the control group, and "B20_Soil" represents the measurement results of the test group. The bar graph in Figure 3 represents the mean value of the dry matter weight, and the error bars represent the standard error. The asterisk in Figure 3 indicates "p<0.05". "p" is the p-value, which represents a statistically significant difference.

[0042] As shown in Figure 3, the dry matter weight of the above-ground portion in the control group was 50.89 ± 6.47 mg, while in the test group it was 87.9 ± 9.58 mg. Furthermore, the dry matter weight of the underground portion in the control group was 9.38 ± 1.16 mg, while in the test group it was 16.03 ± 2.65 mg. A Tukey-Kramer test with a significance level of 5% was performed on these results, and it was found that the dry matter weight of both the underground and above-ground portions of the grown komatsuna seedlings was statistically significantly increased by the application of the B20 strain suspension.

[0043] Thus, the second test revealed that strain B20 has the property of promoting the growth of komatsuna (Japanese mustard spinach). Therefore, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) have the advantage of easily promoting the growth of plants of the genus Brassica. Furthermore, agricultural compositions containing strain B20 are thought to easily promote the growth of plants of the family Brassicaceae. Moreover, agricultural compositions containing strain B20 are thought to easily promote the growth of plants of the order Brassicales.

[0044] <Third Exam> Next, we will explain the contents and results of the third trial concerning strain B20. The third trial was conducted to determine whether strain B20 has a growth-promoting effect on alfalfa, a plant belonging to the genus Medicago in the family Fabaceae, order Fabales. Specifically, in the third trial, a glass bead growing medium was prepared by sterilizing quartz wool glass beads (manufactured by Toshin Riko Co., Ltd.) with a diameter of 2.5 to 3.5 mm in an autoclave and then spreading them in a seedling box. Then, 10 control groups were prepared in which a plant medium mixture (without the bacterial suspension of strain B20) with the composition shown in the "Control Group" section of Table 1 below was applied to the glass bead growing medium, and then 10 alfalfa seeds that had been germinated separately were sown. In addition, 10 test groups were prepared in which a plant medium mixture (with the bacterial suspension of strain B20) with the composition shown in the "Test Group" section of Table 1 below was applied to the glass bead growing medium, and then 10 alfalfa seeds that had been germinated separately were sown.

[0045] [Table 1]

[0046] The composition of "MS medium" in Table 1 is as shown in Table 2 below. In Table 2, "M" represents the molar concentration (mol / L).

[0047] [Table 2]

[0048] After preparation of each test plot and control plot, alfalfa seedlings were grown for two weeks. Subsequently, the dry weight of the above-ground part, the dry weight of the underground part, and the total length of the lateral roots (hereinafter referred to as "total lateral root length") of the grown alfalfa seedlings were measured in each test plot and control plot.

[0049] Figure 4 shows the results of the third experiment. The upper graph in Figure 4 shows the measurement results of the dry matter weight of the above-ground part of the alfalfa seedlings, and the middle graph in Figure 4 shows the measurement results of the dry matter weight of the underground part of the alfalfa seedlings. The lower graph in Figure 4 shows the measurement results of the total lateral root length of the alfalfa seedlings. In Figure 4, "No Bacillus" represents the measurement results of the control group, and "With Bacillus" represents the measurement results of the experiment group. The bar graphs in the upper and middle sections of Figure 4 represent the average value of the dry matter weight, and the bar graph in the lower section of Figure 4 represents the average value of the total lateral root length, with error bars representing the standard error.

[0050] As shown in the upper and middle sections of Figure 4, the dry matter weight of the above-ground portion in the control group was 1.70 ± 0.10 mg, while in the test group it was 3.08 ± 0.52 mg. Furthermore, the dry matter weight of the underground portion in the control group was 0.52 ± 0.09 mg, while in the test group it was 1.67 ± 0.16 mg. A t-test with a significance level of 5% was performed on these results, and it was found that the dry matter weight of both the underground and above-ground portions of the alfalfa seedlings was statistically significantly increased by the application of the B20 strain suspension.

[0051] Furthermore, as shown in the lower part of Figure 4, the total lateral root length was 1.19 ± 0.27 cm in the control group, while it was 5.12 ± 0.54 cm in the test group. A t-test with a significance level of 0.1% was performed on these results, and it was found that the total lateral root length of the alfalfa seedlings was statistically significantly increased by applying the suspension of the B20 strain of fungus.

[0052] Thus, the third test revealed that strain B20 has the property of promoting the growth of alfalfa. Therefore, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) have the advantage of easily promoting the growth of plants of the genus Medicago. Furthermore, agricultural compositions containing strain B20 are thought to easily promote the growth of leguminous plants as well. Moreover, agricultural compositions containing strain B20 are thought to easily promote the growth of leguminous plants as well.

[0053] <Fourth Exam> Next, we will explain the contents and results of the fourth experiment concerning strain B20. The fourth experiment was conducted to determine whether strain B20 has a growth-promoting effect on tomatoes, which are plants belonging to the genus Solanum in the family Solanaceae, order Solanales. Specifically, in the fourth experiment, similar to the third experiment, glass bead cultivation media were prepared by autoclaving 2.5-3.5 mm diameter quartz wool glass beads (manufactured by Toshin Riko Co., Ltd.) and then spreading them in seedling boxes. Then, 10 control groups were prepared in which a plant medium mixture (without the bacterial suspension of strain B20) with the composition shown in the "Control Group" in Table 1 above was applied to the glass bead cultivation media, and then 10 separately germinated tomato seeds were sown. In addition, 10 test groups were prepared in which a plant medium mixture (with the bacterial suspension of strain B20) with the composition shown in the "Test Group" in Table 1 above was applied to the glass bead cultivation media, and then 10 separately germinated tomato seeds were sown.

[0054] After preparation of each test plot and control plot, tomato seedlings were grown for two weeks. Subsequently, the dry weight of the above-ground and below-ground parts of the grown tomato seedlings was measured in each test plot and control plot.

[0055] Figure 5 shows the results of the fourth experiment. The upper graph in Figure 5 shows the measurement results of the dry matter weight of the above-ground parts of the tomato seedlings, and the lower graph in Figure 5 shows the measurement results of the dry matter weight of the underground parts of the tomato seedlings. In Figure 5, "No Bacillus" represents the measurement results of the control group, and "With Bacillus" represents the measurement results of the experiment group. In Figure 5, the bar graphs represent the average value of the dry matter weight, and the error bars represent the standard deviation.

[0056] As shown in Figure 5, the dry matter weight of the above-ground part of the control group was 2.11 ± 0.14 mg, while in the test group it was 2.61 ± 0.18 mg. Similarly, the dry matter weight of the underground part of the control group was 1.3 ± 0.18 mg, while in the test group it was 1.57 ± 0.13 mg. A t-test with a significance level of 5% showed that the dry matter weight of the above-ground part of the tomato seedlings increased statistically significantly with the application of the B20 strain bacterial suspension. On the other hand, the dry matter weight of the underground part of the tomato seedlings did not increase statistically significantly with the application of the B20 strain bacterial suspension, but there was a tendency for it to increase.

[0057] Thus, the fourth test revealed that strain B20 has the property of promoting growth in tomatoes. Therefore, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) have the advantage of easily promoting the growth of plants of the genus Solanum. Furthermore, agricultural compositions containing strain B20 are thought to easily promote the growth of plants of the Solanaceae family. Moreover, agricultural compositions containing strain B20 are thought to easily promote the growth of plants of the Solanales order.

[0058] <Fifth Exam> Next, we will explain the contents and results of the fifth experiment concerning strain B20. The fifth experiment was conducted to examine the effective turbidity of the growth-promoting effect of strain B20 on plants. Specifically, in the fifth experiment, similar to the third experiment, glass beads made of quartz wool (manufactured by Toshin Riko Co., Ltd.) with a diameter of 2.5 to 3.5 mm were sterilized by autoclave and then spread in seedling boxes to create a glass bead cultivation medium. Then, ten control groups were prepared by applying a plant medium mixture (without the bacterial suspension of strain B20) with the composition shown in the "control group" in Table 3 below to the glass bead cultivation medium, and then sowing 10 alfalfa seeds that had been germinated separately. Since the control group did not contain the bacterial suspension of strain B20, the turbidity (OD) of strain B20 was not measured. 600 ) is 0.

[0059] In addition, 60 test plots were prepared in which a plant culture medium mixture (containing a suspension of the B20 strain) with the composition shown in the "Test Plot" column of Table 3 below was applied to a glass bead culture medium, and then 10 alfalfa seeds that had been germinated separately were sown. In the 60 test plots, the turbidity of the B20 strain was adjusted for every 10 plots. Specifically, 10 plots were prepared with a B20 strain turbidity of 0.001, 10 plots with a B20 strain turbidity of 0.005, 10 plots with a B20 strain turbidity of 0.01, 10 plots with a B20 strain turbidity of 0.05, 10 plots with a B20 strain turbidity of 0.1, and 10 plots with a B20 strain turbidity of 0.5.

[0060] [Table 3]

[0061] The composition of "MS medium" in Table 3 is as shown in Table 2 above.

[0062] After preparation of each experimental and control plot, alfalfa seedlings were grown for two weeks. Subsequently, the total lateral root length of the grown alfalfa seedlings was measured in each experimental and control plot.

[0063] Figure 6 shows the results of the fifth experiment. The graph in Figure 6 shows the measurement results of the total lateral root length of the grown alfalfa seedlings. The bar graph in Figure 6 represents the average value of the total lateral root length, and the error bars represent the standard error. In Figure 6, one asterisk indicates "p<0.05", two consecutive asterisks indicate "p<0.01", and three consecutive asterisks indicate "p<0.001". "p" is the p-value, which represents a statistically significant difference, and here it represents the value when compared to the control group where the turbidity of the B20 strain was 0 according to the Welch T Test.

[0064] As shown in Figure 6, the total lateral root length of the alfalfa seedlings was statistically significantly increased when the turbidity of the B20 plant was 0.05 or higher. Furthermore, the total lateral root length of the alfalfa seedlings was statistically significantly increased when the turbidity of the B20 plant was 0.1 or higher, and even more statistically significantly increased when the turbidity of the B20 plant was 0.5 or higher.

[0065] Thus, the fifth test revealed that strain B20 exhibits a growth-promoting effect on alfalfa at a turbidity of 0.05 or higher. Furthermore, it was found that strain B20 exhibits a more pronounced growth-promoting effect on alfalfa at a turbidity of preferably 0.1 or higher. Moreover, it was found that strain B20 exhibits a more pronounced growth-promoting effect on alfalfa at a turbidity of 0.5 or higher.

[0066] Therefore, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) are thought to have the property of promoting growth not only for alfalfa but also for other plants at a turbidity of 0.05 or higher, and are considered to be effective in promoting plant growth.

[0067] <6th Exam> Next, we will explain the contents and results of the sixth test concerning strain B20. The sixth test was to determine whether strain B20 produces auxin, a growth hormone common to plants. Specifically, in the sixth test, strain B20 was cultured in NYAc medium with the composition shown in Table 4 below. The culture supernatant was concentrated 10-fold and then analyzed by LC / MS-MS (liquid chromatography-tandem mass spectrometry). For this analysis, a Cadenza CD-C18 column (manufactured by Intact Co., Ltd.) was used, and an Esquire 3000 spectrometer (manufactured by BRUKER) was used as the analytical instrument. The results of this analysis were then compared with the results of the analysis of indole-3-acetic acid (IAA), a type of auxin.

[0068] [Table 4]

[0069] The composition of "Mineral Basal Medium" in Table 4 is as shown in Table 5 below.

[0070] [Table 5]

[0071] Furthermore, the composition of "Metal-EDTA solution" in Table 5 is as shown in Table 6 below.

[0072] [Table 6]

[0073] Figure 7 shows the results of the sixth test. The upper graph in Figure 7 shows the analysis results for indole-3-acetic acid, and the lower graph in Figure 7 shows the analysis results for the culture supernatant of strain B20. As shown in Figure 7, in the analysis results for the culture supernatant of strain B20, a peak indicated by the arrow was detected at a retention time of 17.7 minutes. Similarly, in the analysis results for indole-3-acetic acid, a peak indicated by the arrow was also detected at a retention time of 17.7 minutes.

[0074] Thus, since the same peak was detected in both the culture supernatant of strain B20 and indole-3-acetic acid, it was found that strain B20 has the property of producing auxin. For this reason, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) are thought to be likely to promote plant growth.

[0075] <7th Exam> Next, we will explain the contents and results of the seventh test concerning strain B20. The seventh test was conducted to determine whether strain B20 exerts a disease-suppressing effect against bacterial wilt. Specifically, in the seventh test, strain B20 was cultured overnight in NY medium or Y medium. Then, the bacterial suspension of the cultured strain B20 was concentrated 100-fold, and 15 μl of this was soaked into paper discs, which were then placed on NYGB medium plates. The composition of NY medium is peptone (5 g / L) and yeast extract (3 g / L). The composition of Y medium is yeast extract (3 g / L). The composition of NYGB medium is peptone (5 g / L), yeast extract (3 g / L), and glycerin (20 ml / L). In addition, a turbidity (OD) of 0.1 was added to the NYGB medium plates. 600 700 μl of a bacterial suspension of the bacterial wilt fungus was applied, and the formation of an inhibition zone was confirmed by further incubation overnight.

[0076] Figure 8 shows the results of the 7th experiment. In Figure 8, "PC" represents the positive control and "NC" represents the negative control. The positive control is a paper disc soaked with 15 μl of streptomycin aqueous solution (250 μg / ml), which is known to have a disease-suppressing effect against bacterial wilt fungus. In Figure 8, "B20 (NY medium)" represents a paper disc soaked with a concentrated suspension of the B20 strain cultured in NY medium, and there are a total of three test groups: "1", "2", and "3". In Figure 8, "B20 (Y medium)" represents a paper disc soaked with a concentrated suspension of the B20 strain cultured in Y medium, and there are a total of three test groups: "1", "2", and "3". In Figure 8, "1", "2", and "3" are represented by the corresponding numbers enclosed in circles.

[0077] As shown in Figure 8, when a concentrated suspension of the B20 strain cultured in Y medium was soaked into a paper disc, an inhibition zone equivalent to that of the positive control was formed. Therefore, it was found that the B20 strain possesses the property of exhibiting a disease-suppressing effect against the bacterial wilt fungus.

[0078] Therefore, agricultural compositions containing microorganisms of the genus Neobacillus (in this case, strain B20) are thought to exert disease-suppressing effects not only against bacterial wilt but also against other plant pathogens. Consequently, agricultural compositions containing microorganisms of the genus Neobacillus exert growth-promoting effects on plants while simultaneously suppressing disease against plant pathogens, making them likely to stably promote plant growth even in environments where plant pathogens are present.

[0079] The embodiments of this disclosure have been described above with reference to the drawings. However, this disclosure is not limited to the embodiments described above, and can be implemented in various forms without departing from its essence.

[0080] This application discloses the following notes. These notes are not intended to limit the present invention.

[0081] (Note 1) An agricultural composition containing microorganisms of the genus Neobacillus.

[0082] (Note 2) The aforementioned microorganism is the species Neobacillus cucumis, as described in Appendix 1 of the agricultural composition.

[0083] (Note 3) The agricultural composition according to Appendix 1 or 2, wherein the microorganism has the property of exhibiting a growth-promoting effect on multiple types of plants.

[0084] (Note 4) The agricultural composition according to any one of the appendices 1 to 3, wherein the microorganism has the property of exhibiting a growth-promoting effect on at least one of Brassicaceae plants and Solanaceae plants.

[0085] (Note 5) The agricultural composition according to any one of the appendices 1 to 4, wherein the microorganism has the property of exhibiting a disease-suppressing effect against plant pathogenic fungi.

[0086] (Note 6) The aforementioned microorganism is strain B20 of the Neobacillus cucumis species (accession number: NITE P-04312), as described in Appendix 2 of the agricultural composition.

[0087] (Note 7) The agricultural composition according to any one of the appendices 1 to 6, wherein the microorganisms have the property of growing in response to the exudate of plant roots.

[0088] (Note 8) The agricultural composition according to any one of the appendices 1 to 7, wherein the microorganism has the property of producing auxin.

[0089] (Note 9) The aforementioned microorganisms have a turbidity of 0.05 or higher (OD 600An agricultural composition described in any one of the appendices 1 to 8, which has the property of exhibiting a growth-promoting effect on plants.

[0090] (Note 10) A seedling product comprising nourishing soil and seedlings cultivated in the aforementioned nourishing soil, A seedling product in which at least one of the seedling and the growing medium is treated with an agricultural composition described in any one of the appendices 1 to 9.

[0091] (Note 11) A method of cultivating plants, A method for cultivating plants, comprising the step of treating the plants with an agricultural composition described in any one of the appendices 1 to 9. [Industrial applicability]

[0092] This invention relates to an agricultural composition containing microorganisms and has industrial applicability.

Claims

1. An agricultural composition containing microorganisms of the genus Neobacillus.

2. The agricultural composition according to claim 1, wherein the microorganism is a species of the genus Neobacillus Cucumis.

3. The agricultural composition according to claim 1, wherein the microorganism has the property of exhibiting a growth-promoting effect on multiple types of plants.

4. The agricultural composition according to claim 1, wherein the microorganism has the property of exhibiting a growth-promoting effect on at least one of Brassicaceae plants and Solanaceae plants.

5. The agricultural composition according to claim 1, wherein the microorganism has the property of exhibiting a disease-suppressing effect against plant pathogenic fungi.

6. The agricultural composition according to claim 2, wherein the microorganism is strain B20 of the Neobacillus cucumis species (accession number: NITE P-04312).

7. The agricultural composition according to any one of claims 1 to 6, wherein the microorganisms have the property of growing in response to the exudate of plant roots.

8. The agricultural composition according to any one of claims 1 to 6, wherein the microorganism has the property of producing auxin.

9. The aforementioned microorganisms have a turbidity of 0.05 or higher (OD ６００ The agricultural composition according to any one of claims 1 to 6, which has the property of exhibiting a growth-promoting effect on plants.

10. A seedling product comprising nourishing soil and seedlings cultivated in the aforementioned nourishing soil, A seedling product in which at least one of the seedling and the growing medium is treated with the agricultural composition described in any one of claims 1 to 6.

11. A method of cultivating plants, A method for cultivating plants, comprising the step of treating the plants with the agricultural composition described in any one of claims 1 to 6.

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