Plant growth promotion material comprising endophyte material and mineral material, plant growth promotion kit, and method for producing plants using the same

Abstract

To provide novel means for promoting the growth of plants.SOLUTION: The present invention provides a plant growth promotion material and a plant growth promotion kit, each comprising an endophyte material including a cultured product of Cephaliophora sp. filamentous fungus and a mineral material, and a method for producing plants using the same.SELECTED DRAWING: Figure 1-1

Description

[Technical field]

[0001] The present invention relates to a plant growth-promoting material containing an endophyte material and a mineral material, a plant growth-promoting kit, and a method for producing plants using the same. [Background technology]

[0002] Endophytes, also known as endophytic fungi, refer to microorganisms that inhabit and symbiotically live within the body of a plant host, thereby providing the host with benefits such as growth promotion and increased stress resistance. Filamentous fungi of the genera Cladosporium sp. and Cephaliophora sp. are known as such endophytes, and in particular, the Cephaliophora sp. strain xsd08001 (hereinafter sometimes referred to as xsd08001 strain) has been reported as a useful microorganism (Patent Document 1).

[0003] A culture of the xsd08001 strain is mixed into soil and used to increase the content of highly functional ingredients in vegetables and other plants (Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2018-174708 A [Patent Document 2] JP 2019-122345 A Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a novel means for promoting plant growth. [Means for solving the problem]

[0006] As a result of intensive studies, the present inventors have first found that when plants are grown in soil containing an endophyte material containing a culture of a filamentous fungus of the genus Cephaliophora sp. and a mineral material, it has an unknown effect of promoting plant growth, and completed the following invention.

[0007] [1] It is a plant growth promoting material containing an endophyte material containing a culture of a filamentous fungus of the genus Cephaliophora sp. The plant growth promoting material contains a mineral material. [2] In the plant growth promoting material according to [1], the mineral material is a composite metal hydroxide having a composition represented by the following formula I. Ca 1-x-y Mg x M 2+ y (OH) 2-nz (A n- ) z (Formula I) [In Formula I, M 2+ represents at least one divalent metal selected from Fe, Mn, Zn and Cu. In Formula I, A n- represents at least one n-valent anion selected from oxyacid ions of B, Mo, P and Se. In Formula I, the range of x is 0 ≦ x < 1. In Formula I, the range of y is 0 < y ≦ 0.5. In Formula I, the range of x + y is 0 < x + y ≦ 1. In Formula I, the range of z is 0 ≦ z ≦ 1. In Formula I, the range of n is 1 ≦ n ≦ 6. In Formula I, the range of nz is 0 ≦ nz ≦ 1.] [3] In the plant growth promoting material according to [2], the above M 2+ is Fe, Mn, Zn and Cu. In the plant growth promoting material, the range of x is 0 ≦ x < 0.4. In the plant growth promoting material, the range of y is 0 < y ≦ 0.3. In the plant growth promoting material, the range of x + y is 0.1 < x + y < 0.5. [4] In the plant growth promoting material according to [2] or [3], the above M 2+ is Fe 0.005~0.1 Mn 0.005~0.1 Zn 0.001~0.1 Cu 0.001~0.05It is represented by the composition. In the above material for promoting plant growth, the range of x is 0.01 < x < 0.4. In the above material for promoting plant growth, the range of y is 0.01 < y ≤ 0.2. In the above material for promoting plant growth, the range of x + y is 0.1 < x + y < 0.5. In the above material for promoting plant growth, A n- is an oxygen acid ion of B and Mo. [5] The material for promoting plant growth according to any one of [1] to [4], wherein the mineral material is contained in an amount of 0.01% by weight to 5% by weight of the total weight of the soil to which it is applied. [6] The material for promoting plant growth according to any one of [1] to [5], wherein the microorganism is Cephaliophora sp. xsd08001 strain.

[0008] [7] A plant growth promoting kit comprising an endophyte material containing a culture of a filamentous fungus of the genus Cephaliophora sp. The plant growth promoting kit contains a mineral material. [8] The plant growth promoting kit according to [7], wherein the mineral material contains a composite metal hydroxide having a composition represented by the following formula I. Ca 1-x-y Mg x M 2+ y (OH) 2-nz (A n- ) z (Formula I) [In Formula I, M 2+ represents at least one divalent metal selected from Fe, Mn, Zn and Cu. In Formula I, A n- represents at least one n-valent anion selected from oxygen acid ions of B, Mo, P and Se. In Formula I, the range of x is 0 ≤ x < 1. In Formula I, the range of y is 0 < y ≤ 0.5. In Formula I, the range of x + y is 0 < x + y ≤ 1. In Formula I, the range of z is 0 ≤ z ≤ 1. In Formula I, the range of n is 1 ≤ n ≤ 6. In Formula I, the range of nz is 0 ≤ nz ≤ 1.] [9] The plant growth promoting kit according to [8], wherein the above M 2+is Fe, Mn, Zn, and Cu. In the above plant growth promoting kit, the range of x is 0 ≦ x < 0.4. In the above plant growth promoting kit, the range of y is 0 < y ≦ 0.3. In the above plant growth promoting kit, the range of x + y is 0.1 < x + y < 0.5. The plant growth promoting kit according to

[10] , [8] or [9], the above M 2+ is Fe 0.005~0.1 Mn 0.005~0.1 Zn 0.001~0.1 Cu 0.001~0.05 is represented by the composition of. In the above plant growth promoting kit, the range of x is 0.01 < x < 0.4. In the above plant growth promoting kit, the range of y is 0.01 < y ≦ 0.2. In the above plant growth promoting kit, the range of x + y is 0.1 < x + y < 0.5. In the above plant growth promoting kit, A n- is the oxyacid ion of B and Mo. The plant growth promoting kit according to

[11] , any one of [7] to

[10] is used such that the above mineral material is contained in an amount of 0.01% by weight to 5% by weight of the total weight of the soil to which it is applied. The plant growth promoting kit according to

[12] , any one of [7] to

[11] is such that the above microorganism is the strain xsd08001 of the genus Cephaliophora sp.

[0009]

[13] A method for producing a plant, A method for producing a plant using soil containing an endophyte material containing a culture of a filamentous fungus of the genus Cephaliophora sp. In the above method for producing a plant, the above soil contains a mineral material.

[14] The method for producing a plant according to

[13] is such that the above mineral material is a composite metal hydroxide having a composition represented by the following formula I. Ca 1-x-y Mg x M 2+ y (OH) 2-nz (A n- ) z (Formula I) [In Formula I, M 2+represents at least one divalent metal selected from Fe, Mn, Zn, and Cu. In Formula I, A n- represents at least one n-valent anion selected from oxyacid ions of B, Mo, P, and Se. In Formula I, the range of x is 0 ≦ x < 1. In Formula I, the range of y is 0 < y ≦ 0.5. In Formula I, the range of x + y is 0 < x + y ≦ 1. In Formula I, the range of z is 0 ≦ z ≦ 1. In Formula I, the range of n is 1 ≦ n ≦ 6. In Formula I, the range of nz is 0 ≦ nz ≦ 1.]

[15]

[14] The method for producing a plant described in

[14] is the above M 2+ is Fe, Mn, Zn, and Cu. In the method for producing the above plant, the range of x is 0 ≦ x < 0.4. In the method for producing the above plant, the range of y is 0 < y ≦ 0.3. In the method for producing the above plant, the range of x + y is 0.1 < x + y < 0.5.

[16]

[14] or

[15] The method for producing a plant described in

[14] or

[15] is the above M 2+ is Fe 0.005~0.1 Mn 0.005~0.1 Zn 0.001~0.1 Cu 0.001~0.05 represented by the composition of. In the method for producing the above plant, the range of x is 0.01 < x < 0.4. In the method for producing the above plant, the range of y is 0.01 < y ≦ 0.2. In the method for producing the above plant, the range of x + y is 0.1 < x + y < 0.5. In the method for producing the above plant, A n- is an oxyacid ion of B and Mo.

[17]

[13] -

[16] The method for producing a plant according to any one of

[13] -

[16] is such that the above mineral material is contained in an amount of 0.01 wt% to 5 wt% of the total weight of the soil to which it is applied.

[18]

[13] -

[17] The method for producing a plant according to any one of

[13] -

[17] is such that the above microorganism is Cephaliophora sp. xsd08001 strain.

Advantages of the Invention

[0010] When a plant is grown by adopting the present invention, the growth of the plant is promoted.

Brief Description of the Drawings

[0011] [Figure 1-1] FIG. 1-1 shows the fresh weight (g) of the above-ground parts of lettuce grown in Experiment 1 (Example 1, Comparative Examples 1 to 6). [Figure 1-2] FIG. 1-2 shows the appearance of lettuce grown in Experiment 1 (Example 1, Comparative Examples 1 to 6). [Figure 2-1] FIG. 2-1 shows the fresh weight (g) of the above-ground parts of mini lettuce grown in Experiment 2 (Example 2, Comparative Examples 7 to 12). [Figure 2-2] FIG. 2-2 shows the appearance of mini lettuce grown in Experiment 2 (Example 2, Comparative Examples 7 to 12). [Figure 3-1] FIG. 3-1 shows the fresh weight (g) of the above-ground parts of mini lettuce grown in Experiment 3 (Example 3, Comparative Examples 13 to 18). [Figure 3-2] FIG. 3-2 shows the appearance of mini lettuce grown in Experiment 2 (Example 3, Comparative Examples 13 to 18). [Figure 4] FIG. 4 shows the amounts (mg) of ingredients (potassium, zinc, vitamin C, vitamin K) contained per 100 g of above-ground parts of lettuce grown in Experiment 4 (Example 4, Comparative Examples 19 to 21). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present invention will be described in detail below with reference to specific embodiments. However, the present invention is not limited to the following embodiments, and can be embodied in any form without departing from the spirit of the present invention.

[0013] One aspect of the present embodiment relates to a plant growth-promoting material comprising an endophyte material containing a culture of a filamentous fungus of the genus Cephaliophora, and a mineral material.

[0014] Moreover, one aspect of the present embodiment relates to a plant growth promotion kit including an endophyte material containing a culture of Cephaliophora sp. filamentous fungus and a mineral material. In the plant growth promotion kit, the endophyte material and the mineral material may be packaged separately. In addition, the plant growth promotion kit may be packaged in a mixed state of the endophyte material and the mineral material. The plant growth promotion kit may further include soil, for example, culture soil.

[0015] Moreover, one aspect of the present embodiment relates to a method for producing a plant, which uses an endophyte material including a culture of a filamentous fungus of the genus Cephaliophora, and soil including a mineral material.

[0016] [I. Endophyte Materials] In the present disclosure, the term "endophyte material" includes a culture of a filamentous fungus that can function as an endophyte, such as Cephaliophora sp., Cladosporium sp., Fusarium sp., Exophiala sp., Lecanicillium sp., and Penicillium sp. Endophytes are also called endophytic fungi, and refer to microorganisms that live and coexist in the body of a plant host, thereby providing the host with benefits such as growth promotion and increased stress resistance. The "endophyte material" that can be applied to this embodiment includes a culture of a filamentous fungus of the genus Cephaliophora, and preferably includes a culture of the Cephaliophora sp. xsd08001 strain. In another embodiment, the endophyte material may contain one or more of the above filamentous fungi in addition to the Cephaliophora filamentous fungus.

[0017] Examples of Cladosporium fungi include Cladosporium cladosporioides, Cladosporium delicatulum, Cladosporium perangustum, Cladosporium tenuissimum, Cladosporium allicinum, Cladosporium colombiae, Cladosporium halotolerans, and Cladosporium parahalotolerans.

[0018] An example of a Fusarium fungus is Fusarium solani.

[0019] An example of a filamentous fungus of the genus Exophiala is Exophiala jeanselmei.

[0020] An example of a Penicillium fungus is Penicillium citrinum.

[0021] In one embodiment, the endophytic material has a particle size distribution of 50% of the cumulative particle size (D 50 ) is 100 to 900 μm and the cumulative 90% particle size (D 90 The above-mentioned filamentous fungal culture having a particle size of 1000 to 5000 μm can be applied to this embodiment.

[0022] [KK] The Cephaliophora sp. xsd08001 strain that can be used in this embodiment was deposited at the National Institute of Technology and Evaluation (NITE) on March 6, 2017, and is available under the accession number NITE P-02438.

[0023] [Particle size] The particle size of the culture of filamentous fungi as an endophyte material that can be used in this embodiment is as follows, but is not limited to these. The cumulative 10% particle size (D10) in the volumetric particle size distribution of the culture of filamentous fungi used in this embodiment is, for example, 50 to 400 μm, particularly 100 to 350 μm, more preferably 150 to 350 μm, and even more preferably 200 to 300 μm. The cumulative 50% particle size (D 50 The cumulative 90% particle diameter (D 90 ) is, for example, 1000 to 5000 μm, in particular, 1100 to 4000 μm, more preferably 1200 to 3000 μm, and even more preferably 1200 to 2000 μm. For example, in one embodiment, the cumulative 50% particle diameter (D 50 ) is 200 to 800 μm, and the cumulative 90% particle diameter (D 90 ) is 1000 to 3000 μm. The particle size can be measured by any method such as a laser diffraction method, a scattering method, an imaging method, a light transmission centrifugal sedimentation method, a sedimentation method, an electrical resistance method, a specific surface area method, or a sieve passing method. The particle size can be measured using an appropriate particle size measuring instrument, for example, a commercially available instrument such as LMS-2000e or LMS-3000 (Seishin Enterprise Co., Ltd.). Since the culture is dried and then pulverized, it is preferable to use a dry measurement method for the particle size.

[0024] In one embodiment, the culture of the xsd08001 strain is obtained by subjecting the xsd08001 strain to solid culture. In another embodiment, the culture of the xsd08001 strain is obtained by subjecting the xsd08001 strain to liquid culture and then to solid culture. In these embodiments, the particles of the culture of the xsd08001 strain may be obtained by a process including drying the solid culture obtained by solid culture and grinding the dried solid culture.

[0025] [Liquid culture] In this specification, "liquid culture" refers to culturing a strain using a liquid medium. The process of performing such liquid culture may be referred to as a "liquid culture process." Subjecting the xsd08001 strain to liquid culture refers to inoculating the strain into a liquid medium and culturing it in the liquid medium. The strain may be in the form of a seed culture medium carried by a medium. An example of the medium is agar. In order to avoid contamination by various bacteria, inoculation is preferably performed under sterile conditions.

[0026] As used herein, the term "liquid culture" refers to a liquid culture obtained as a result of liquid culture. A liquid medium can be prepared, for example, by adding various additives to the liquid to adjust the composition to be suitable for culture. An example of the liquid is water. Examples of additives added to the liquid medium include sugars, minerals, nitrogen sources, vitamins, organic acids, inorganic acids, organic bases, and inorganic bases. Preferably, the liquid medium contains sugars and a nitrogen source. An example of the nitrogen source contained in the liquid medium is peptone.

[0027] Examples of sugars include, but are not limited to, one or more selected from glucose, galactose, fructose, maltose, sucrose, lactose, oligosaccharides, and glycerol. The upper limit of the sugar is not limited, but the total amount of sugars relative to the liquid medium is usually 60 g / L or less, particularly 50 g / L or less, further 40 g / L or less, and particularly 30 g / L or less. If the liquid medium contains too many nutrients such as sugars, the mycelium concentration will be too high. On the other hand, the lower limit of the sugar is not limited, but the total amount of sugars relative to the liquid medium is usually 0.5 g / L or more, particularly 1 g / L or more, further 5 g / L or more, and particularly 10 g / L or more. If the nutrients such as sugars are too few, the mycelium will not grow sufficiently. For example, in one embodiment, the sugar is glucose, and is added in an amount of 20 g / L of medium.

[0028] The minerals may be inorganic salts, such as alkali and alkaline earth metal salts, and salts of other metals, but are not limited thereto. Examples of such inorganic salts include one or more salts selected from sulfates, phosphates, carbonates, chlorides, alkali metal oxides, molybdates, selenites, and halides. One or more salts may be used. The upper limit of the salts is not limited, but the total amount of salts in the liquid medium, excluding the amount contained in yeast extract and peptone, is usually 10.0 g / L or less, particularly 8.0 g / L or less, more preferably 5.0 g / L or less, and particularly preferably 2.0 g / L or less. This is because, as described above, if the liquid medium contains too many salts as nutrients, the mycelium concentration becomes too high. On the other hand, the lower limit of the salts is not limited, but the total amount of salts in the liquid medium, excluding the amount contained in the yeast extract, is usually 0.01 g / L or more, more preferably 0.05 g / L or more, even more preferably 0.1 g / L or more, and particularly preferably 0.25 g / L or more. This is because if the liquid medium contains too few minerals such as salts, the mycelium does not grow sufficiently. For example, in one embodiment, magnesium sulfate is added as salts in an amount of 0.5 g / L of medium, and potassium dihydrogen phosphate is added in an amount of 0.1 g / L of medium.

[0029] The nitrogen source may be one or more nitrogen sources selected from yeast extract, protein hydrolysates, and proteins. Examples of protein hydrolysates include peptone. Adding yeast extract and / or peptone to a liquid medium can add nutrients including protein hydrolysates such as amino acids and peptides, proteins, and salts. The amount of nitrogen source relative to the liquid medium is not limited, but for example, the upper limit of the nitrogen source is usually 30.0 g / medium L or less, particularly 20.0 g / medium L or less, further 15.0 g / medium L or less, and particularly 10.0 g / medium L or less, as the total amount of nitrogen sources such as yeast extract and peptone. The lower limit of the nitrogen source is usually 0.05 g / medium L or more, particularly 0.1 g / medium L or more, further 0.5 g / medium L or more, and particularly 1.0 g / medium L or more, as the total amount of nitrogen sources such as yeast extract and peptone. For example, the amount of yeast extract relative to the liquid medium is not limited, but the upper limit of yeast extract is usually 20.0 g / L or less, particularly 15.0 g / L or less, further 10.0 g / L or less, and particularly preferably 5.0 g / L or less. The lower limit of yeast extract is usually 0.01 g / L or more, particularly preferably 0.05 g / L or more, further 0.1 g / L or more, and particularly preferably 0.5 g / L or more. The amount of peptone relative to the liquid medium is not limited, but the upper limit of peptone is usually 10.0 g / L or less, particularly preferably 8.0 g / L or less, further 5.0 g / L or less, and particularly preferably 2.0 g / L or less. The lower limit of peptone is usually 0.01 g / L or more, particularly preferably 0.05 g / L or more, further 0.1 g / L or more, and particularly preferably 0.5 g / L or more. As above, if the nutrients are too high, the mycelium concentration will be too high, while if the nitrogen source is too low, the mycelium will not grow sufficiently. For example, in one embodiment, yeast extract is added in an amount of 2.0 g / L of medium, and peptone is added in an amount of 1.0 g / L of medium. Furthermore, the pH may be appropriately adjusted by adding an appropriate acid or base, if necessary.

[0030] The liquid culture can be carried out by a suitable culture means. For example, a container such as an Erlenmeyer flask can be used, and the culture can be carried out while stirring with a stirring device such as a stirrer. In addition, it is preferable that the culture conditions of the liquid culture are adjusted by a temperature control device or the like. For the liquid culture, one or more devices selected from a vibration device, a humidity measuring device, a pH control device, a turbidity measuring device, a light control device, a specific gas concentration measuring device, and a pressure measuring device may be used as necessary. The specific gas concentration measuring device may be capable of measuring, for example, O2 and CO2 as the specific gas. For the liquid culture, a silicon plug may be used optionally from the viewpoint of preventing contamination. For the liquid culture, aeration stirring culture, shaking culture, stationary culture, or the like may be appropriately carried out. For example, in one embodiment, a liquid medium and a stirrer are placed in an Erlenmeyer flask, the mouth of the flask is sealed with a silicon plug, and stirring culture with breathability is carried out.

[0031] The duration of liquid culture is not limited, but may be adjusted to a desired concentration, for example, an average colony count of usually 1×10 per 1 L of liquid culture. 3 cfu / L of medium or more, especially 1×10 4 cfu / L of medium or more, even 1×10 5 cfu / L of medium or more, particularly preferably 1×10 6 cfu / L of medium or more, e.g., about 5 x 10 6 The culture can be completed when the number of cfu / L of medium is reached. For example, the upper limit of the liquid culture period is usually 8 days or less, more preferably 6 days or less, even more preferably 5 days or less, and particularly preferably 4 days or less. The lower limit of the liquid culture period is usually 6 hours or more, more preferably 12 hours or more, even more preferably 18 hours or more, and particularly preferably 1 day or more. If the liquid culture period is too long, the mycelium concentration will be too high. On the other hand, if the liquid culture period is too short, the mycelium will not elongate sufficiently. For example, in one embodiment, if the liquid culture period is 2 to 4 days, for example 3 days, a liquid culture with a concentration appropriate for solid culture following liquid culture can be obtained.

[0032] [Solid culture] In this specification, "solid culture" refers to culturing a strain using a solid culture medium containing a solid medium and water. The process of performing such solid culture may be referred to as a "solid culture process". A solid culture refers to a culture obtained as a result of solid culture in which a strain is cultured in a solid culture medium. For example, bran, soybean pulp, bamboo powder, sawdust, rice husk, bagasse, cellulose powder, cellobiose, coffee grounds, and starch can be used as the solid culture medium. The water used in solid culture may contain, for example, various additives that contribute to the growth of the strain. Examples of additives include antibiotics that can suppress the growth of bacteria other than the Cephaliophora sp. xsd08001 strain used in this embodiment.

[0033] The solid culture medium can be prepared by adding water to the solid culture medium and mixing. The moisture content of the solid culture medium is not limited, but for example, the upper limit of the moisture content is usually 85% by weight or less, particularly 80% by weight or less, further 75% by weight or less, and particularly 70% by weight or less is preferable. The lower limit of the moisture content is usually 30% by weight or more, particularly 40% by weight or more, further 45% by weight or more, and particularly 50% by weight or more is preferable. If the moisture content of the solid culture medium is too high, the medium becomes too watery and sufficient gaps are not formed in the medium, so that growth is not promoted. On the other hand, if the moisture content of the solid culture medium is too low, sufficient moisture for culture cannot be secured. For example, in one embodiment, when the moisture content of the solid culture medium is 50 to 70% by weight, for example, 60% by weight, a solid culture having a mycelium concentration appropriate as an endophyte material can be obtained.

[0034] In this specification, subjecting a liquid culture to solid culture refers to inoculating a solid culture medium with the liquid culture to perform solid culture. For inoculation, for example, a method of inoculating using a micropipette or other tool, or a method of sterilizing the container containing the strain by at least one of the following measures, namely, heating the mouth of the container with a burner, followed by decanting and direct inoculation, can be adopted. Regardless of the method used, inoculating a liquid culture into a solid culture medium allows the liquid culture to be distributed throughout the solid culture medium. This allows the mycelium to grow more uniformly throughout the solid culture medium, resulting in a more uniform culture, which in turn is believed to result in culture particles with a more uniform particle size.

[0035] Solid culture can be carried out by a suitable culture means. For example, solid culture can be carried out by using a container such as a plastic bag and allowing the container to stand. In addition, for solid culture, it is preferable to adjust the culture conditions using a temperature control device, a humidity measuring device, etc. If necessary, one or more devices selected from a stirring device, a vibration device, a pH adjusting device, a turbidity measuring device, a light control device, a specific gas concentration measuring device, and a pressure measuring device may be used. The specific gas concentration measuring device may be capable of measuring, for example, O2 and CO2 as the specific gas. For solid culture, it is preferable to use a disposable container from the viewpoint of preventing contamination, and measures such as closing the mouth of the bag may be taken as appropriate. Stirring culture, shaking culture, static culture, etc. may be carried out as appropriate. For example, in one embodiment, a disposable bag equipped with a ventilation filter is used, and static culture is carried out by closing the mouth of the bag.

[0036] The duration of solid culture is not limited, but may be adjusted to a desired concentration, for example, an average colony count of usually 1×10 per gram of solid culture. 3 ~1×10 7 cfu / g, among which 5 × 10 3 ~5×10 6 cfu / g, or even 1×10 4 ~1×10 6 cfu / g, particularly preferably 5×10 4 ~5×10 5cfu / g, e.g., about 1×10 5 The culture can be completed when the cell count reaches cfu / g. For example, the upper limit of the solid culture period is usually 20 days or less, preferably 18 days or less, more preferably 15 days or less, and particularly preferably 12 days or less. The lower limit of the solid culture period is usually 5 days or more, more preferably 6 days or more, more preferably 7 days or more, and particularly preferably 8 days or more. If the solid culture period is too long, the risk of contamination by various bacteria increases. In addition, if the solid culture period is too long, it hinders efficient production of the endophyte material. On the other hand, if the solid culture period is too short, the mycelium does not grow sufficiently. For example, in one embodiment, if the solid culture is performed for 8 to 12 days, for example, 10 days, a solid culture having a mycelium concentration appropriate for an endophyte material can be obtained.

[0037] [Humidity] The humidity during liquid culture is not particularly limited and can be any humidity. On the other hand, the humidity during solid culture is not limited, but the upper limit of the humidity for solid culture is usually 100% RH or less, particularly 95% RH or less, more preferably 90% RH or less. The lower limit of the humidity for solid culture is usually 55% RH or more, particularly 60% RH or more, more preferably 65% ​​RH or more. In one embodiment, the humidity can be 60 to 80% RH, for example, 70% RH.

[0038] [temperature] In both liquid culture and solid culture, the upper limit of temperature is usually 40° C. or lower, preferably 35° C. or lower, more preferably 30° C. or lower, and particularly preferably 25° C. or lower. The lower limit of temperature in liquid culture and solid culture is usually 5° C. or higher, preferably 10° C. or higher, more preferably 15° C. or higher, and particularly preferably 20° C. or higher. In one embodiment, the culture can be carried out at 20 to 25° C., for example, 25° C.±1° C.

[0039] [Sterilization] From the viewpoint of preventing contamination, it is preferable to sterilize the liquid medium, solid medium, and / or solid culture medium by any known means, such as filtration sterilization, autoclave sterilization, boiling sterilization, radiation sterilization, sodium hypochlorite, or ozone treatment. Each operation, such as inoculation, is preferably performed in a sterile atmosphere. For example, in the case of a solid culture medium, an appropriate amount of moisture can be added to the solid culture medium before sterilization.

[0040] [Culture] The ratio of the culture of filamentous fungi in the endophyte material applicable to this embodiment is not limited. For example, the upper limit of the ratio of the culture of filamentous fungi, for example, the xsd08001 strain of the genus Cephaliophora in the endophyte material is 100% by weight or less, 95% by weight or less, 90% by weight or less, or 80% by weight or less. The lower limit of the ratio of the culture of the xsd08001 strain in the material is 10% by weight or more, 20% by weight or more, or 30% by weight or more. In one aspect, the material may be composed of a culture of the xsd08001 strain. The method for producing a culture of a filamentous fungus may include subjecting a filamentous fungus to liquid culture and then to solid culture. The method for producing a culture of a filamentous fungus may also include drying the solid culture obtained by solid culture and pulverizing the dried solid culture. In addition, the method may also include classifying the pulverized product as necessary. For classification, for example, a sieve can be used. In this specification, drying the solid culture obtained in the solid culture step to produce a dried solid culture is sometimes referred to as a "drying step". Furthermore, crushing the dried solid culture is sometimes referred to as a "crushing step".

[0041] Drying can be carried out using general-purpose equipment such as an air conditioner or a dehumidifier so as to achieve the following culture concentrations, for example. For example, drying can be carried out by leaving the culture in an environment of 25°C and 50% RH for 3 days. Grinding can be carried out using general-purpose grinders such as a food processor, coffee mill, or pepper mill.

[0042] In the method for producing a culture of a filamentous fungus, since the mycelium extends uniformly within the culture, it is possible to obtain particles of the culture with a desired particle size by simply drying and pulverizing the solid culture. In one embodiment, however, the cumulative 50% particle size (D 50 ) is 100 to 900 μm and the cumulative 90% particle size (D 90 In the present specification, the cumulative 50% particle diameter (D 50 ) is 100 to 900 μm and the cumulative 90% particle size (D 90 The process of adjusting the particle size to 1000 to 5000 μm is sometimes referred to as an “adjustment process.” The adjustment process includes a pulverization process.

[0043] Since the material has a uniform particle size and the fungus is uniformly distributed, it can stably function as an endophyte in plants, which is preferable from the viewpoint of plant growth and quality improvement. In addition, the uniform particle size makes it easy to handle, and is advantageous for tableting, granulation, packaging, transportation, etc. By using the strain of this embodiment, a material in which the fungus is uniformly distributed can be suitably obtained even by solid culture alone. Furthermore, when liquid culture is used in combination, the mycelium extends throughout the medium, making the concentration of the fungus even more uniform.

[0044] The concentration of the solid culture obtained in this embodiment is, for example, usually 1 × 10 as the average number of colonies per 1 g of solid culture. 3 ~1×10 7 cfu / g, among which 5 × 10 3 ~5×10 6 cfu / g, or even 1×10 4 ~1×10 6 cfu / g, particularly preferably 5×10 4 ~5×10 5 cfu / g, e.g., about 1×10 5 The culture concentration in the material of this embodiment is, for example, 1 × 10 average colony count per 1 g of dry solid culture. 5 ~1×10 9 cfu / g, among which 5 × 10 5 ~5×10 8cfu / g, or even 1×10 6 ~1×10 8 cfu / g, particularly preferably 5×10 6 ~5×10 7 cfu / g. The average number of colonies per gram of dry solid culture remains unchanged before and after the crushing process.

[0045] [II. Mineral Materials] [element] In this disclosure, the term "mineral material" refers to a material for adding inorganic compounds to soil, which is used to promote crop growth. The mineral material applied in this embodiment contains at least one of magnesium (Mg), iron (Fe), zinc (Zn), manganese (Mn), copper (Cu), molybdenum (Mo), boron (B), calcium (Ca), sulfur (S), and nickel (Ni). Various mineral materials are commercially available so that they can be selected according to the plant to be grown and the purpose.

[0046] The mineral materials used in this embodiment may be materials containing, for example, boron (B), iron (Fe), manganese (Mn), zinc (Zn), copper (Cu) and molybdenum (Mo). The inclusion of these elements can suppress metabolic disorders such as photosynthesis or nitrate reduction in plants, and deficiencies resulting from each component.

[0047] The mineral materials used in this embodiment may include, for example, materials containing magnesium (Mg), calcium (Ca), boron (B), iron (Fe), manganese (Mn), zinc (Zn), copper (Cu) and molybdenum (Mo). The inclusion of these elements can supply elements essential for plant growth.

[0048] Each element contained in the mineral material applied in this embodiment may be in a form that can be utilized by plants or endophytes. The mineral material may be a mixture of compounds containing each element. The mineral material may be a complex containing multiple elements.

[0049] Examples of the forms of elements contained in mineral materials include: Boron (B): Water-soluble boron, citric acid-soluble boron, and insoluble boron can be used, and water-soluble boron is preferred. Examples include boric acid, sodium borate, sodium perborate, boron oxide, and boron trifluoride. Iron (Fe): For example, iron(III) oxide, ferrous sulfate, ferrous sulfate, and ferrous sulfate hydrate. Manganese (Mn): Water-soluble manganese, chelate-soluble manganese, and insoluble manganese can be used, with water-soluble manganese being preferred. Examples include manganese oxide, manganese sulfate, and manganese sulfate hydrate. Zinc (Zn): For example, zinc oxide, zinc stearate, zinc phosphate, zinc sulfate, zinc sulfate hydrate. Copper (Cu): For example, copper stearate, copper sulfate, and copper sulfate hydrate. Molybdenum (Mo): Water-soluble molybdates are preferred. For example, alkali metal salts of molybdic acid include ammonium molybdate, sodium molybdate, potassium molybdate, and lithium molybdate. Ammonium molybdate is also included. Molybdenum oxide. Magnesium (Mg): Water-soluble magnesium, chloro-soluble magnesium, and insoluble magnesium can be used, with water-soluble magnesium being preferred. Examples include magnesium chloride and magnesium sulfate. · Calcium (Ca): Examples include calcium chloride, calcium acetate, monocalcium phosphate, and calcium sulfate.

[0050] [Optimal amounts of elements in soil] The optimal values ​​for each element contained in soil can be determined by combining various test values, empirical values, etc.

[0051] Examples of optimum values ​​(ppm by mass) in soil for each of the above elements are as follows: Boron: 0.4~3.5 Manganese: 7~35 Iron: 10~150 ·Copper: 1~5 Zinc: 10~40 Molybdenum: 0.09~6

[0052] [Specific examples of mineral materials] In one aspect, the mineral material applied to this embodiment may be, for example, a composite metal hydroxide having a composition represented by the following formula I described in JP 2019-122345 A (Patent Document 2).

[0053] Ca 1-x-y Mg x M 2+ y (OH) 2-nz (A n- ) z (Formula I) [wherein, M 2+ represents at least one divalent metal selected from Fe, Mn, Zn and Cu, and A n- represents at least one n-valent anion selected from the group consisting of oxygen acid ions of B, Mo, P, and Se, and the ranges of x, y, z, and n are 0≦x<1, 0 <y≦0.5、0<x+y≦1、0≦z≦1、1≦n≦6、0≦nz≦1である。]

[0054] In one embodiment, in the composite metal hydroxide represented by the above formula (I), M 2+ The metal may be at least one of Zn, Fe, Mn and Cu. Other metals include Cr. 3+ and / or Ni 2+ Preferred M 2+ is preferably Zn, Fe, Mn or Cu. 2+ is Fe 0.005~0.1 Mn 0.005~0.1 Zinc 0.001~0.1 Cu 0.001~0.05 The composition may be in the range of

[0055] In one embodiment, in the composite metal hydroxide represented by the above formula (I), A n-It can be at least one or more n-valent anions selected from oxyacid ions of B, Mo, P, and Se. Examples of the oxyacid ions of B, Mo, P, and Se include, for example, HBO3 2- , BO3 3- , B4O7 2- , MoO4 2- , Mo2O7 2- , H2PO4 - , HPO4 2- , PO4 3- , SeO3 2- , SeO4 2- and the like, but it is not limited to this.

[0056] In one aspect, in the composite metal hydroxide represented by the above (Formula I), the range of x is 0 ≦ x < 1, preferably 0 ≦ x < 0.4, more preferably 0.01 < x < 0.4.

[0057] In one aspect, in the composite metal hydroxide represented by the above (Formula I), the range of y is 0 < y ≦ 0.5, preferably 0 < y ≦ 0.3, more preferably 0.01 < y ≦ 0.2.

[0058] In one aspect, in the composite metal hydroxide represented by the above (Formula I), the range of x + y is 0 < x + y ≦ 1, preferably 0.1 < x + y < 0.5.

[0059] In one aspect, in the composite metal hydroxide represented by the above (Formula I), the range of z is 0 ≦ z ≦ 1, preferably 0 ≦ z ≦ 0.5.

[0060] In one aspect, in the composite metal hydroxide represented by the above (Formula I), the range of n is 1 ≦ n ≦ 6.

[0061] In one aspect, in the composite metal hydroxide represented by the above (Formula I), the range of nz is 0 ≦ nz ≦ 1, preferably 0 ≦ nz ≦ 0.5.

[0062] In one aspect, the composite metal hydroxide represented by the above (Formula I) is Ca, Mg, and M2+ The compound can be produced by dissolving a water-soluble salt of the compound, such as a sulfate, chloride, or nitrate, in water, and co-precipitating the resulting solution with an equivalent or greater amount of an alkali, such as sodium hydroxide or potassium hydroxide. The preferred pH and temperature of the coprecipitation reaction are 10 to 14 and 20 to 50°C, respectively. After the coprecipitation reaction, the compound is filtered, washed with water, dehydrated, dried, and pulverized. When a molybdate, phosphate, or selenate is added, it is added after the coprecipitation reaction and before filtration, and the mixture is stirred. When a borate is added, it is added to the cake after washing with water and dehydration, and mixed. The drying atmosphere may be air, but is preferably a non-oxidizing atmosphere. By drying in a non-oxidizing atmosphere, oxidation of Fe and Mn can be reduced.

[0063] In another reaction method, calcium hydroxide, which functions as both an alkali and a Ca source, and Mg and M 2+ A method of reacting the above with a mixed aqueous solution of the above can also be used.

[0064] In one embodiment, the composite metal hydroxide represented by the above formula (I) may be used so as to be contained in an amount of 0.01% to 5% by weight of the total weight of the soil to which it is applied, for example, 0.01% to 5% by weight, 0.05% to 3% by weight, or 0.1% to 1% by weight.

[0065] [III. Soil] In one embodiment, the soil is not particularly limited and may be any soil that can be used for cultivating normal crops, such as commercially available culture soil or soil accumulated on a desired land.

[0066] [IV. Plants to be cultivated and production methods] The plants to be cultivated in this embodiment may be, for example, trees, flowers, vegetables, or fruit trees. For example, when the plants to be cultivated in this embodiment are cultivated as vegetables for fresh produce cultivation, that is, cultivation for the purpose of market shipment or consumption, the plants may be leafy vegetables such as cabbage, Chinese cabbage, spinach, etc., whose leaves are edible. The plants may also be root vegetables such as radish, turnip, carrot, and green onion, whose roots and stems are edible. The plants may also be fruit vegetables such as cucumbers, green peppers, and eggplants, which are intended for fruit production.

[0067] In one aspect, the vegetables or fruit trees to be cultivated by this embodiment include, for example, Chenopodiaceae crops such as spinach, Swiss chard, and sugar beet, Asteraceae crops such as lettuce, lettuce, chrysanthemum, and burdock, Brassicaceae crops such as cabbage, broccoli, Chinese cabbage, radish, and turnip, Liliaceae crops such as onion and leek, Apiaceae crops such as carrot, celery, and mitsuba, Solanaceae crops such as tomato, eggplant, bell pepper, capsicum, torvum, acanthus, and tobacco, Cucurbitaceae crops such as cucumber, melon, watermelon, pumpkin, and kanpyo, Poaceae crops such as sweet corn, legume crops such as pea, broad bean, kidney bean, and soybean, and Rosaceae crops such as strawberry, apple, loquat, plum, cherry, peach, and pear. In particular, the application of this embodiment to Asteraceae crops and Chenopodiaceae crops is suitable.

[0068] In one aspect, examples of flowers to be cultivated in this embodiment include horticultural or ornamental plants such as roses, Asteraceae plants such as chrysanthemums, dandelions, and thistles, and Liliaceae plants such as lilies and tulips.

[0069] To produce a plant by applying this embodiment, the plant growth-promoting material is applied to soil in which the plant is to be grown, and the plant is grown according to conventional farming methods. EXAMPLES

[0070] The present invention will now be described in more detail with reference to examples, although the present invention is not limited thereto.

[0071] <Materials and methods used> 1. Preparation of endophyte materials The strains used as endophytes were Cephaliophora sp. xsd08001 (referred to as "Cephaliophora.sp" in this example), deposited with the National Institute of Technology and Evaluation (NITE) under the accession number NITE P-02438, and Cladosporium cladosporioides isolate A1S1-1 (hereinafter referred to as "Cladosporium"), which is known as a common endophyte. Each strain was made into a material in the following manner.

[0072] 2. Materialization method 2-1. Preculture A preculture medium having the following composition was prepared and used to preculture Cephaliophora sp. and Cladosporium.

[0073] [Table 1]

[0074] 2-2.Main culture Cephaliophora sp. and Cladosporium were cultured according to the following procedure. (1) Commercially available okara powder was mixed with ion-exchanged water to a moisture content of 50%, and this was used as the culture bed. (2) The culture medium in (1) was sterilized in an autoclave. (3) The bacteria precultured in 2-1 was inoculated onto the culture bed. (4) The opening of the culture bed bag was sealed with a sealer, and the bag was left to stand in an environment of 25°C and a humidity of 70% or higher.

[0075] The number of colonies per gram of the culture medium after crushing and mixing well is 1x10 5 The incubation was completed when the incubation temperature reached 100°C.

[0076] 2-3. Drying and grinding (1) The bag containing the cultured culture medium in 2-2 was torn open, and the culture medium was crushed on a tray. (2) The mushroom bed was allowed to dry. (3) The dried mushroom bed was crushed in a grinder.

[0077] 2-4. Preparation of mineral materials The mineral materials used were those described in JP 2019-122345 A. Specifically, they were prepared by the following procedure.

[0078] A mixed aqueous solution of calcium chloride, magnesium chloride, zinc chloride, ferrous nitrate, manganese nitrate and cupric nitrate (Ca 2+ = 0.616 mol / L, Mg 2+ =0.25mol / L, Fe 2+ = 0.06 mol / L, Mn 2+ = 0.045 mol / L, Zn 2+ =0.025mol / L, Cu 2+ Aqueous sodium hydroxide solutions (0.004 mol / L) and 4 mol / L were fed into a 2.5-liter overflow reactor at flow rates of 100 ml / min and 50 ml / min, respectively, using a metering pump. The pH of the reactor was kept at about 13 and the temperature at about 40°C while stirring with a chemical stirrer. Coprecipitation reaction was continuously carried out for about 30 minutes. 0.0064 mol of sodium molybdate dihydrate was added to 1 mol of solids contained in the suspension obtained by overflow, and the mixture was stirred for 10 minutes. After that, the mixture was filtered under reduced pressure and washed with water to obtain a cake. 0.026 mol of boric acid was added to 1 mol of solids contained in the cake and mixed. The cake was placed in a hot air dryer and dried at about 100°C for 6 hours. After cooling to about 40°C, the cake was taken out of the dryer and pulverized with an atomizer to obtain a mineral material applicable to the present invention. The X-ray diffraction pattern of the obtained powder was measured, and the diffraction pattern was almost exclusively that of calcium hydroxide. The chemical composition of this powder was analyzed by ICP and found to be as follows:

[0079] Ca 0.616 Mg 0.25 Fe 0.06 Mn 0.045 Zinc 0.025 Cu0.004 (OH) 1.9352 (HBO3 2- ) 0.026 (MoO4 2- ) 0.0064

[0080] <Experiment 1> Crop cultivation test: lettuce Crop cultivation tests were conducted under the following test conditions. (1) Materials used Culture soil used: Takii vegetable soil (per 1L: Nitrogen (N): 280mg, Phosphorus (P): 320mg, Potassium (K): 300mg) 670g / pot Sample seeds: Lettuce, Okayama lettuce (Takii Seeds Co., Ltd.) Three lettuce plants were planted in each pot, and three pots were cultivated in each test plot. The temperature inside the greenhouse during the test period was 18 to 25°C. Sample endophytes: Cephaliophora.sp (Example 1, Comparative Example 4), Cladosporium cladosporioides isolate A1S1-1 (Comparative Examples 5 and 6) Cultivation period: 46 days

[0081] (2) Soil preparation The cultivation soil was prepared by adding each material listed in Table 2 to the culture soil. Table 2 shows the amount of each material added per pot.

[0082] [Table 2]

[0083] (3) Seeding and cultivation methods The prepared cultivation soil for each test plot was placed in a pot, seeds were sown, and water was applied uniformly to each test plot. After sowing, when it was time to harvest, the above-ground parts of the lettuce were harvested and their fresh weights were measured.

[0084] (4) Results The fresh weight (average value) of the above-ground parts of lettuce is shown in Figure 1-1. The state of the lettuce at the time of harvest is also shown in a photograph (Figure 1-2). As is clear from Figure 1-1, it was suggested that when the Cephaliophora endophyte material and the mineral material were mixed into the culture soil (Example 1), the growth of lettuce was promoted synergistically.

[0085] <Experiment 2> Crop cultivation test: Mini lettuce 1. Test conditions Crop cultivation tests were conducted under the following conditions: (1) Materials used Culture soil used: Takii vegetable soil (per 1L: Nitrogen (N): 280mg, Phosphorus (P): 320mg, Potassium (K): 300mg) 670g / pot Sample species: Takii-grown semi-head mini lettuce Manoa (Takii Seeds Co., Ltd.) (hereinafter referred to as "mini lettuce") Three mini lettuce plants were planted in each pot, and three pots were cultivated in each experimental area. The temperature inside the greenhouse during the experiment was 18 to 25°C. Sample endophytes: Cephaliophora.sp (Example 2, Comparative Example 10), Cladosporium cladosporioides isolate A1S1-1 (Comparative Example 11, Comparative Example 12) Cultivation period: 42 days

[0086] (2) Soil preparation The cultivation soil was prepared by adding each material shown in Table 3 to the culture soil. Table 3 shows the amount of each material added per pot.

[0087] [Table 3]

[0088] (3) Seeding and cultivation methods The prepared cultivation soil for each test plot was placed in a pot, seeds were sown, and water was applied uniformly to each test plot. After sowing, when it was time to harvest, the above-ground parts of the mini lettuce were harvested and the fresh weight was measured.

[0089] (4) Results The fresh weight (average value) of the aboveground parts of mini lettuce is shown in Figure 2-1. The state at the time of harvest is also shown in a photograph (Figure 2-2). As is clear from Figure 2-1, it was suggested that the growth of mini lettuce was promoted synergistically when the endophytic material of the Cephaliophora genus and the mineral material were mixed into the culture soil (Example 2).

[0090] <Experiment 3> Crop cultivation test: Spinach 1. Test conditions Crop cultivation tests were conducted under the following conditions: (1) Materials used Culture soil used: Takii vegetable soil (per 1L: Nitrogen (N): 280mg, Phosphorus (P): 320mg, Potassium (K): 300mg) 670g / pot Sample species: Takii Hybrid Spinach Okame (Takii Seed Co., Ltd.) (hereinafter referred to as "Spinach") Five spinach plants were planted in each pot, and three pots were cultivated in each experimental area. The temperature in the greenhouse during the experiment was 18 to 25°C. Sample endophytes: Cephaliophora.sp (Example 3, Comparative Example 16), Cladosporium cladosporioides isolate A1S1-1 (Comparative Example 17, Comparative Example 18) Cultivation period: 42 days

[0091] (2) Soil preparation The materials listed in Table 4 were added to the culture soil to prepare the cultivation soil. Table 4 shows the amount of each material added per pot.

[0092] [Table 4]

[0093] (3) Seeding and cultivation methods The prepared cultivation soil for each test plot was placed in a pot, seeds were sown, and water was applied uniformly to each test plot. After sowing, when it was time to harvest, the above-ground parts of the spinach were harvested and their fresh weights were measured.

[0094] (4) Results The fresh weight (average value) of the above-ground part of spinach is shown in Figure 3-1. The state at the time of harvest is also shown in a photograph (Figure 3-2). As is clear from Figure 3-1, it was suggested that when the endophyte material of the Cephaliophora genus and the mineral material were mixed into the culture soil (Example 3), the growth of spinach was promoted synergistically.

[0095] <Experiment 4> Crop cultivation test: lettuce Crop cultivation tests were conducted under the following test conditions. (1) Materials used Culture soil used: Takii vegetable soil (per 1L: Nitrogen (N): 280mg, Phosphorus (P): 320mg, Potassium (K): 300mg) 670g / pot Sample seeds: Lettuce, Okayama lettuce (Takii Seeds Co., Ltd.) Two lettuce plants were planted in each pot, and three pots were cultivated in each test area. The temperature inside the greenhouse during the test period was 18 to 25°C. Sample endophyte: Cephaliophora.sp (Example 4, Comparative Example 21) Cultivation period: 32 days

[0096] (2) Soil preparation The cultivation soil was prepared by adding each material shown in Table 5 to the culture soil. Table 5 shows the amount of each material added per pot.

[0097] [Table 5]

[0098] (3) Seeding and cultivation methods The prepared cultivation soil for each test plot was placed in a pot, seeds were sown, and water was applied uniformly to each test plot. After sowing, the above-ground parts of the lettuce were harvested when it was time to harvest. The Japan Food Research Laboratories was asked to analyze the various components of the harvested lettuce.

[0099] (4) Results The content of each component per 100g of lettuce is shown in Figure 4. As is clear from Figure 4, it was suggested that when the Cephaliophora endophyte material and the mineral material were mixed into the culture soil, the nutrients (potassium, zinc, vitamin C, and vitamin K) contained in lettuce increased.

Claims

1. an endophytic material comprising a culture of a filamentous fungus of the genus Cephaliophora; A plant growth promoting material including a mineral material.

2. The mineral material is a composite metal hydroxide having a composition represented by the following formula I: Ca １－ｘ－ｙ Mg ｘ M ２＋ ｙ (OH) ２－ｎｚ (A ｎ－ ) ｚ (Formula I) [wherein, in formula I, M ２＋ represents at least one divalent metal selected from Fe, Mn, Zn and Cu, A ｎ－ represents at least one n-valent anion selected from the oxygen acid ions of B, Mo, P, and Se, and the ranges of x, y, z, and n are 0≦x<1, 0<y≦0.5, 0<x+y≦1, 0≦z≦1, 1≦n≦6, and 0≦nz≦1. The plant growth promoting material according to claim 1,

3. 3. The plant growth-promoting material according to claim 1 or 2, wherein the filamentous fungus is a Cephaliophora sp. xsd08001 strain.

4. an endophytic material comprising a culture of a filamentous fungus of the genus Cephaliophora; A plant growth promotion kit including mineral materials.

5. A method for producing a plant, comprising the steps of: A method for producing plants using soil containing an endophyte material including a culture of a filamentous fungus of the genus Cephaliophora and a mineral material.

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