JP6192162B2 - Tree planting material and its manufacturing method - Google Patents

Description

  The present invention relates to a greening material using microorganisms symbiotic with plants and a method for producing the same.

  Conventionally, symbiotic microorganisms such as ectomycorrhizal fungi are allowed to coexist in the host plant, and it is known that effects such as promoting the growth of the host plant, improving resistance to environmental stress, and improving fruit quality can be obtained. (See, for example, Patent Document 1). However, when such symbiotic microorganisms are applied directly to soil, it is difficult to obtain a stable effect due to differences in soil characteristics. In addition, in order for symbiotic microorganisms such as those described above to have a symbiotic relationship with plants in the soil, a certain degree of bacterial density is required, but usually they originally live in the soil rather than the applied symbiotic microorganisms. Since the density of the other microorganisms is much higher, it was difficult to fully exert the functions of the symbiotic microorganisms. For this reason, in developing a greening material utilizing the action of such symbiotic microorganisms, it is necessary to find a material suitable for the growth and stable maintenance of the symbiotic microorganism and to hold the symbiotic microorganisms at a high density in the material.

  By the way, plant charcoal such as charcoal and bamboo charcoal has been used for a long time as a soil conditioner in horticulture and agriculture. However, it is difficult to grow symbiotic microorganisms such as ectomycorrhizal fungi and arbuscular mycorrhizal fungi at high density on such plant charcoal, and the plant charcoal retains the symbiotic microorganisms to constitute a greening material. However, the effect could not be fully exhibited.

JP 2007-74986 JP JP 2012-80829 A

  Therefore, the present inventor has already found that the above problem can be solved by maintaining the symbiotic microorganisms after adjusting the pH of the plant carbide to an appropriate value, and has developed a new planting material (see Patent Document 2). . The planting material is obtained by holding ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi in a plant having a pH of 5.0 to 8.0 by adding an acidic liquid. In this way, by adding an acidic liquid to the plant carbide so as to have a pH of 5.0 to 8.0, usually, the plant charcoal whose chemical characteristics are weakly alkaline to alkaline is favored under weakly acidic to neutral conditions. It can be made suitable as a material for retaining ectomycorrhizal fungi and arbuscular mycorrhizal fungi.

  However, the greening material still has room for improvement in terms of mycorrhizal fungus retention density and effects when used on soil.

  As a result of intensive studies, the inventor causes an ectomycorrhizal fungus and / or arbuscular mycorrhizal fungus of a specific strain to be retained in a plant charcoal added with an acidic liquid and adjusted to an appropriate pH. Thus, the present inventors have found that an effect superior to that of the prior art can be obtained and completed the present invention.

The planting material according to the present invention maintains the mycelium of ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi in a plant having a pH of 5.0 to 8.0 by adding an acidic liquid to the charcoal. Which consists of
The ectomycorrhizal fungus or the arbuscular mycorrhizal fungus may be selected from the group consisting of the Ichiuchi family Chichiawatake 1140-TK44 strain (NITE AP-01658), the xylem family Shakashimeji 273-TS49 strain (NITE AP-01659), or the genus Glomus genus KANSO. It is any one strain or multiple strains of -19 strains.

  According to the greening material according to the present invention, it is possible to retain mycorrhizal fungi at a higher density than conventional greening materials, and it is possible to stably exhibit superior effects when used in soil. It becomes.

The graph which shows the evaluation result of the microbial cell growth in Example 1. FIG. The graph which shows the evaluation result of the fungal body growth of the mushroom at each pH in Example 2. FIG. The graph which shows the evaluation result of the cell growth of Shakashimeji at each pH in the Example. The graph which shows the evaluation result of the microbial cell proliferation in Example 3. FIG. The graph which shows the evaluation result of the microbial cell growth of arbuscular mycorrhizal fungi in each pH in Example 4. The graph which shows the evaluation result of the plant nurturing | growth effect by the Chichiawatake material in Example 5. The graph which shows the evaluation result of the plant breeding effect by Shakashimeji material in Example 6. The graph which shows the evaluation result of the plant breeding effect by the arbuscular mycorrhizal fungi material in Example 7. It is a graph which shows the evaluation result of the root system reproduction | regeneration effect in Example 8, Comprising: (a) shows the amount of fine roots per 1L of soil, (b) shows mycorrhiza formation rate. It is a graph which shows the evaluation result of the root system regeneration effect in Example 9, Comprising: (a) shows the amount of fine roots per 1L of soil, (b) shows mycorrhiza formation rate.

  The greening material according to the present invention has an ectomycorrhizal fungus and / or a pH of 5.0 to 8.0 (more preferably pH 6.0 to 7.0) by adding an acidic liquid to the plant carbide. Or arbuscular mycorrhizal fungi, wherein the ectomycorrhizal fungus or the arbuscular mycorrhizal fungus is the Ichiuchi family Chichiawatake 1140-TK44 strain (deposit agency: National Institute of Technology and Evaluation) Patent Microbiology Deposit Center, Receipt Number: NITE AP-01658, Receipt Date: July 19, 2013), Kishimeji Shachashimeji 273-TS49 strain (Deposit Organization: National Institute of Technology and Evaluation, Patent Microorganism Deposit Center, Receipt Number) : NITE AP-01659, date of receipt: July 19, 2013), and any of the strains of the genus Glomus genus KANSO-19 It is characterized by being one or multiple shares.

  In addition, since the said Glomus genus KANSO-19 strain was refused deposit by the independent administrative institution product evaluation technology base mechanism patent microorganism deposit center for a technical reason, this applicant guarantees for sale.

  The above ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi can be selected from one or more suitable ones according to the type of plant and soil characteristics to which the greening material of the present invention is applied. It is desirable to use it.

  Note that the pH is measured by the pH electrode method. Specifically, first, a 10 g sample of a carbide sample in terms of dry weight is weighed and placed in a 200 mL Erlenmeyer flask. Next, add 50 mL of distilled water, put a rubber stopper and shake for 30 minutes. Finally, transfer the contents of the Erlenmeyer flask to a 100 mL beaker and measure the pH using a pH meter.

  The plant carbide is obtained by heating and carbonizing a plant body. As the plant carbide in the present invention, general charcoal having neutral to alkaline pH characteristics can be suitably used. In addition, although the pH of a plant carbide changes with carbonization temperature, it is desirable to use the charcoal produced | generated at the carbonization temperature of 400 to 800 degreeC as a plant carbide in this invention. In general, charcoal that is distributed as an agricultural and horticultural material is manufactured at the carbonization temperature and has neutral to alkaline pH characteristics. Therefore, the greening material according to the present invention can be easily manufactured using commercially available charcoal. In addition, as plant charcoal in the present invention, charcoal of various plants such as bamboo charcoal can be used in addition to so-called charcoal obtained by carbonizing wood such as oak, beech, oak and kunugi.

  The mycorrhiza is mainly formed into fine roots having a diameter of 2 mm or less. Therefore, the plant carbide preferably has a particle size of 2 mm to 10 mm (more desirably 2 mm to 6 mm). As a result, the rooting of fine roots as described above can be induced, and mycorrhizal formation efficiency can be improved. The particle size of the plant carbide can be measured by a sieving method.

  As the acidic liquid in the present invention, it is desirable to use one or more of organic acids such as acetic acid, citric acid or vinegar, vegetable juice, and fruit juice. Thereby, the plant carbide can be adjusted to pH characteristics suitable for the growth of symbiotic microorganisms, and the effect of promoting the growth of mycelium can be obtained. In addition, as said vegetable juice, commercially available vegetable juice etc. can be used, for example.

  The acidic liquid may be added to the plant carbide by any method. For example, it is possible to employ a method such as spraying an acidic liquid on the surface of the plant carbide using a sprayer or immersing the plant carbide in an acidic liquid.

  In addition, the greening material according to the present invention is, for example, after adding a predetermined amount (for example, a volume equal to the amount of plant carbide) of sand and mixing the above-mentioned acidic liquid to plant carbide and mixing the mixture with high-pressure sterilization. Then, a host plant such as alfalfa or sorghum previously infected with a symbiotic microorganism (for example, arbuscular mycorrhizal fungi) in this sterilized mixture is planted, and a predetermined temperature (for example, about 20 to 30 ° C.) for a predetermined period (for example, about 30 to 40). Day) and can be produced by fully extending the mycelium in the mixture.

  Still further, the greening material according to the present invention comprises adding the acidic liquid and a liquid medium containing nutrients necessary for the survival of symbiotic microorganisms to plant charcoal, and then subjecting the mixture to a sterilized mixture in advance. A certain amount of symbiotic microorganisms (eg, ectomycorrhizal fungi) cultured using a predetermined medium are inoculated, cultured at a predetermined temperature (for example, 20 to 30 ° C.) for a predetermined period (for example, about 30 to 40 days), and the plant It can also be produced by fully extending the mycelium in a mixture of carbide and liquid medium.

In addition, as said culture medium, the culture medium containing a nutrient required for growth of a symbiotic microorganism is used. For example, PDB (Potato Dextrose Broth) medium, MMN (Modified Melin-Norkrans) medium, etc. can be used as the liquid medium added to the plant carbide. Moreover, as a culture medium for cultivating a symbiotic microorganism beforehand before making it hold | maintain to the said plant carbide, PDA (Potato Dextrose Agar) culture medium, MMN culture medium, etc. can be used, for example. The PDB medium can be prepared, for example, by dissolving 4 g of potato starch and 20 g of dextrose in 1000 mL of purified water. The PDA medium can be prepared, for example, by dissolving 4 g of potato starch, 20 g of dextrose and 20 g of agar in 1000 mL of purified water. The MMN medium is, for example, glucose 10.0 g, ammonium tartrate 1.0 g, KH ₂ PO ₄ 0.5 g, MgSO ₄ .7H ₂ O 0.15 g, CaCl ₂ .2H ₂ O 0.05 g, 1% FeCl ₃ solution 1 .2 mL, 0.1% 0.1% thiamine hydrochloride solution 0.1 mL, malto extract (Difco) 3.0 g, yeast extract (Difco) 2.0 g are dissolved in purified water 1000 mL and adjusted to pH 5.5. Can be prepared.

The high-pressure sterilization means sterilization with high-temperature and high-pressure steam (so-called autoclave sterilization). For example, the treatment is performed with superheated steam at 121 ° C. at 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ). It is desirable. The treatment time varies depending on the target substance to be sterilized. When cultivating soil such as a mixture of plant carbide and sand, the processing time is about 30 minutes because there are few problems with soil alteration and sterilization is difficult. When medium such as PDB medium or PDA medium is subjected to high-pressure sterilization treatment, if the treatment time is lengthened, the medium components are altered, so the treatment time is about 15 minutes.

  The mycelium elongated inside the greening material by the above-described method can immediately start an activity if there is a plant root, and can quickly establish a symbiotic relationship. However, on the other hand, there is a possibility that the symbiotic ability may be affected by drying and high temperature. Therefore, it is desirable to add a spore suspension of the symbiotic microorganism to the greening material produced by the above method. Spores have the problem that it takes time to germinate and symbiotic with plants, but they have the advantage of being able to withstand dryness and high temperatures and maintain long-term symbiotic ability. Therefore, by holding both mycelium and spores in the plant carbide as described above, it is possible to obtain a greening material that complements the disadvantages of both and has both immediate effect and stability of effect.

  Further, by the above-described method, two or more kinds of greening materials each holding different symbiotic microorganisms are produced, and by mixing these, a greening material comprising two or more kinds of symbiotic microorganisms is produced. Good. Such a mixed material can be suitably used, for example, when a plurality of types of plants symbiotic with different symbiotic microorganisms are mixed in one place.

  The tree planting material according to the present invention as described above can be suitably used as a tree-restoring material for activating a tree or fruit tree whose tree has declined. In addition, the greening material according to the present invention can be used as a substitute for a fertilizer or a part of a fertilizer in a greening construction or a landscaping work for planting a seedling, or in raising a flower bud, a vegetable, or a fruit tree. In any case, the greening material according to the present invention is applied by being mixed or embedded in the soil where the target plant (that is, the plant to be symbiotic with the microorganism) is planted.

  When the greening material according to the present invention is applied to black pine, charcoal temperature of 400 ° C. to 800 ° C. and particle size of 2 to 6 mm are used as the plant carbide, and ectomycorrhizal fungi are used as symbiotic microorganisms. desirable. Moreover, when applying the greening material according to the present invention to Japanese red pine, charcoal temperature of 400 ° C. to 600 ° C. and particle size of 2 to 6 mm are used as the plant carbide, and ectomycorrhizal fungi are used as symbiotic microorganisms. It is desirable. Furthermore, when the greening material according to the present invention is applied to a tree of the genus Sakura, charcoal temperature of 400 to 800 ° C. and charcoal of 2 to 6 mm are used as the plant carbide, and arbuscular mycorrhiza as symbiotic microorganisms. It is desirable to use bacteria.

[Proliferation and materialization of ectomycorrhizal fungi]
First, 500 mL of a 40% strength vinegar solution was mixed with 1 L of charcoal (pH 9.0) having a particle size of 2 to 5 mm and air-dried to prepare charcoal having a pH adjusted to 6.0. Next, 500 mL of MMN liquid medium was added to the charcoal to impregnate the charcoal with nutrients necessary for the growth of ectomycorrhizal fungi. At that time, 10 mL of unsalted vegetable juice (manufactured by Kagome Co., Ltd.) was centrifuged and added to the MMN medium in advance. Thereafter, the charcoal containing the liquid medium was autoclaved at 121 ° C. and 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ) for 15 minutes. On the other hand, 1140-TK44 (NITE AP-01658) colonies of 1140-TK44 strain (Suillus granulatus), which is an ectomycorrhizal fungus of Iguchi family previously plated on MMN medium, were cut into 5 mm square pieces and subjected to the sterilization treatment. Inoculated into charcoal containing fresh liquid medium and cultured at 28 ° C. A greening material was obtained by culturing this for about 1 month until the charcoal was sufficiently covered with white mycelium. Hereinafter, this is referred to as 1140-TK44 strain material.

  Moreover, the greening material which hold | maintained Shakoshimeji (Lyophyllum fumosum) 273-TS49 strain (NITE AP-01659) which is an ectomycorrhizal fungus of the xylem family was produced by the same method. This is hereinafter referred to as 273-TS49 strain material.

  Furthermore, as a comparative example, by the same method, a greening material (hereinafter referred to as 1140 strain material) in which another strain (1140 strain) of Chichiwatake was retained, and another strain (273 strain) of Shakashimeji were retained. A greening material (hereinafter referred to as 273 strain material) was prepared. Further, as another comparative example, the commercially available horticultural culture medium is used instead of charcoal, and a normal MMN medium not added with vinegar and vegetable juice is used, and the above-mentioned Tichiaiwatake 1140 strain or Shakashimeji 273 strain is obtained by the same procedure as described above. A greening material was produced. Moreover, the greening material which hold | maintained the said Tichiaiwatake 1140 strain or the said Shakashimeji 273 strain by the same procedure using the normal MMN culture medium which does not add vinegar and vegetable juice as another comparative example was produced.

  FIG. 1 shows the results of evaluating the cell growth effect of the ectomycorrhizal fungi in the materials of the 1140-TK44 strain, the 273-TS49 strain and the comparative examples. Here, the cell growth effect was evaluated by comparing the dry weight of the cells contained in each material. The dry weight of the cells was measured by drying the material prepared above with a drier and subtracting the dry weight of charcoal or horticultural soil previously measured from the dry weight.

  As a result, among the materials of the comparative examples, greening materials prepared using charcoal without adding vinegar and vegetable juice ("1140 strain + normal charcoal" and "273 strain + normal charcoal" in the figure) In 1140 strain material and 273 strain material, compared with the planting material using horticultural soil ("1140 strain + horticultural soil" and "273 strain + horticultural soil" in the figure) It was confirmed that a high amount of cells can be obtained. Further, it was confirmed that the 1140-TK44 strain material and the 273-TS49 strain material, which are the greening materials according to the present invention, have a better cell growth effect than the 1140 strain material and the 273 strain material. .

[Influence of material pH]
Preparation of 1140-TK44 strain material (Example) and 1140 strain material (Comparative example), and 273-TS49 strain material (Example) and 273 strain material (Comparative example) of Shakashimeji using various pH charcoal In the same manner as in Example 1, the cell growth effect was evaluated. In addition, adjustment of pH adjusts pH by mixing 500 mL of vinegar solutions of 20%, 30, 40, and 50% concentration with 1 mL of charcoal (pH 9.0) having a particle size of 2 to 5 mm and air-dried. Specifically, charcoal adjusted to 5, 6, 7, 8, and 9 was prepared.
As a result, as shown in FIG. 2 and FIG. 3, a high cell growth effect was observed at pH 5.0 to 8.0, particularly pH 6.0 to 7.0, and the amount of cells decreased at pH 9.0. The tendency to do was recognized. Furthermore, in the material using Tichiaiwatake, the greening material of the example (1140-TK44 strain material) showed higher cell mass than the greening material of the comparative example (1140 strain material) at all pH except pH 9.0. . Moreover, in the material using Shakashimeji, the greening material of the example (273-TS49 strain material) showed higher cell mass than the greening material of the comparative example (273 strain material) at all pHs.

[Arbuscular mycorrhizal fungus body growth and materialization]
First, a high-pressure sterilizer was applied to a cell tray (cell tray # 72, manufactured by Tocan Kosan Co., Ltd.) having a capacity of 15 mL per plug seedling at 121 ° C. and 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ). 7 mL of river sand sterilized for 30 minutes by (Tomy Seiko Co., Ltd., BS-325) was added, and then 2 mL of spore solution of arbuscular mycorrhizal fungus Glomus genus KANSO-19 was added and further sterilized. 3 mL of the dried river sand was added and seeded with alfalfa seeds. This was properly cultivated and managed under conditions of an air temperature of 20 to 30 ° C. and irrigated in a timely manner. In addition, Peters' liquid fertilizer (Daiichi Gardening Co., Ltd., NPK: 25-5-20) was sufficiently irrigated every week after germination. After continuing cultivation for 30 days, alfalfa fine roots were collected, decolorized with a 10% aqueous potassium hydroxide solution, stained with trypan blue staining solution, and the symbiotic status of arbuscular mycorrhizal fungi was confirmed using a microscope.

Next, 500 mL of a 40% strength vinegar solution was mixed with 1 L of charcoal (pH 9.0) having a particle size of 2 to 5 mm and air-dried to prepare charcoal having a pH adjusted to 6.0. Thereafter, the charcoal and an equal volume of river sand were mixed, and the mixture was autoclaved for 30 minutes at a temperature of 121 ° C. and 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ). Using this mixture as soil, alfalfa previously grown on a cell tray was transplanted. This was properly cultivated and managed under conditions of an air temperature of 20 to 30 ° C. and irrigated in a timely manner. In addition, Peters' liquid fertilizer (Daiichi Horie, NPK: 25-5-10) was sufficiently irrigated every week after germination. Cultivation was continued for 30 to 40 days, and it was confirmed that arbuscular mycorrhizal fungi were symbiotic to the fine roots of alfalfa by the method described above. Thereafter, irrigation was stopped, the plant body was killed to promote spore formation in the culture soil, and the culture soil was sufficiently dried to obtain a greening material retaining arbuscular mycorrhizal fungi. This is hereinafter referred to as KANSO-19 strain material.

  In addition, as a comparative example, a revegetation material holding another strain of arbuscular mycorrhizal fungus (Glomus sp. OG-105 strain) and another strain of arbuscular mycorrhiza (Glomus sp. KANSO) A greening material retaining -6 strain) was produced by the same method as described above. These are hereinafter referred to as OG-105 strain material and KANSO-6 strain material, respectively.

  Further, as a comparative example, a greening material holding each strain of the arbuscular mycorrhizal fungi was prepared by using a commercially available horticultural soil instead of charcoal and by the same procedure (but not adding vinegar). Furthermore, as another comparative example, a greening material holding each strain of the arbuscular mycorrhizal fungi was prepared by the same procedure as described above using charcoal to which vinegar was not added.

  The evaluation result of the cell growth effect of the arbuscular mycorrhizal fungi in the above-mentioned KANSO-19 strain material (Example) and the greening material according to each comparative example is shown in FIG. In addition, here, the cell growth effect was evaluated by comparing the number of spores per 100 g of dry soil (dry material).

  As a result, all of the KANSO-19 strain material, the OG-105 strain material, and the KANSO-6 strain material were greening materials using the horticultural soil ("KANSO-19 strain + horticultural soil" in the figure, "OG-105"). "Strain + Horticultural soil" and "KANSO-6 strain + Horticultural soil") and greening materials using charcoal without vinegar ("KANSO-19 strain + normal charcoal" in the figure, "OG-105 strain + It was confirmed that the number of spores was higher than that of “ordinary charcoal” and “KANSO-6 strain + ordinary charcoal”), and a high cell growth effect was obtained. Furthermore, in the greening material ("KANSO-19 strain material" in the figure) according to the present invention, the greening material ("OG-105 strain material in the figure") produced using other strains of Arbuscular mycorrhizal fungi And “KANSO-6 strain material”), a further excellent cell growth effect was confirmed.

[Influence of material pH]
KANSO-19 strain material (Example), OG-105 strain material (Comparative example), and KANSO-6 strain material (Comparative example) of Tichiaiwatake using various pH charcoal, and similar to Example 3 above Thus, the cell proliferation effect was evaluated. In addition, adjustment of pH adjusts pH by mixing 500 mL of vinegar solutions of 20%, 30, 40, and 50% concentration with 1 mL of charcoal (pH 9.0) having a particle size of 2 to 5 mm and air-dried. Specifically, charcoal adjusted to 5, 6, 7, 8, and 9 was prepared. As a result, as shown in FIG. 5, a high cell growth effect is observed at pH 5.0 to 8.0, especially pH 6.0 to 7.0, and the amount of the cells tends to decrease at pH 9.0. Admitted. Further, at any pH, the planting material according to the present invention (KANSO-19 strain material) has a higher cell growth effect than the greening materials of the comparative examples (OG-105 strain material and KANSO-6 strain material). It was done.

[Effects of black pine grown with ectomycorrhizal fungi]
After seeding black pine seeds on vermiculite (manufactured by Nittai Co., Ltd.) that had been subjected to dry heat sterilization at 180 ° C. for 8 hours, seedlings with aseptic roots of uniform size were cultivated by timely irrigation. Next, seedling seedlings of the black pine are placed in a mixed culture medium of equal amounts of river sand and red crust that has been subjected to high-pressure sterilization for 30 minutes at 121 ° C. and 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ). At that time, 1140-TK44 strain material (Example) or 1140 strain material (Comparative Example), which is a material of the bamboo shoot produced in Example 1, is applied to the planting hole by 10 mL for one seedling seedling. did. When this seedling was cultivated while applying Peters liquid fertilizer (NPK: 25-5-10) every two weeks, it was larger than the seedling cultivated only with fertilizer (control group) on the 60th day of cultivation. I was able to get a seedling. In particular, the 1140-TK44 strain material, which is a greening material according to the present invention, was confirmed to have an even better growth effect than the 1140 strain material, which is a comparative example. FIG. 6 shows the dry weight of the above-ground part and underground part of seedlings on the 60th day of cultivation. In addition, the figure has shown the average value of the result of having repeated the said test twice by making the number of seedlings of each test section into ten.

[Growing effect of Pinus densiflora with ectomycorrhizal fungi]
After seeding red pine seeds on vermiculite (manufactured by Nittai Co., Ltd.) that had been subjected to dry heat sterilization at 180 ° C. for 8 hours, seedlings with aseptic roots with uniform sizes were grown by irrigating timely. Next, the seedlings of Pinus densiflora seedlings were mixed in an equal volume mixed soil of river sand and red crust that had been subjected to high-pressure sterilization for 30 minutes under the conditions of 121 ° C. and 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ). In this case, 273-TS49 strain material (Example) or 273 strain material (Comparative Example), which is the material of Shakashimeji prepared in Example 1, is applied to the planting hole by 10 mL per seedling seedling. did. When this seedling was cultivated while applying Peters liquid fertilizer (NPK: 25-5-10) every two weeks, it was larger than the seedling cultivated only with fertilizer (control group) on the 60th day of cultivation. I was able to get a seedling. In particular, in the 273-TS49 strain material, which is a greening material according to the present invention, a further superior growth effect was confirmed as compared with the 273 strain material, which is a comparative example. FIG. 7 shows the dry weight of the above-ground part and the underground part of seedlings on the 60th day of cultivation. In addition, the figure has shown the average value of the result of having repeated the said test twice by making the number of seedlings of each test section into ten.

[Growing effect of miscellaneous cherry by Arbuscular mycorrhizal fungus material]
After seeding Kasumi cherry seeds on vermiculite (manufactured by Nittai Co., Ltd.) that had been subjected to dry heat sterilization at 180 ° C. for 8 hours, seedlings of aseptic roots with uniform sizes were cultivated by irrigating timely. Next, the seedling seedlings of Kasumi cherry tree were put into an equal volume mixed soil of river sand and red crust that had been subjected to high-pressure sterilization for 30 minutes under the conditions of 121 ° C. and 11.8 × 10 ⁴ Pa (1.2 kgf / cm ² ). At that time, 10 mL of the KANSO-19 strain material, the OG-105 strain material, or the KANSO-6 strain material prepared in Example 3 was applied to the planting hole for each of the seedlings. When this seedling was cultivated while applying Peters' liquid fertilizer (NPK: 25-5-10) every two weeks, on the 60th day of cultivation, only the fertilizer was applied in any section where the greening material was applied. A larger seedling than the cultivated seedling (control) was able to be obtained. In particular, in the district where the KAANSO-19 strain material, which is the greening material according to the present invention, is applied, the seedlings are more excellent than the OG-105 strain material, which is the greening material of the comparative example, or the kanso-6 strain material. The training effect was confirmed. FIG. 8 shows the dry weight of the above-ground part and the underground part of seedlings on the 60th day of cultivation. In addition, the figure has shown the average value of the result of having performed the said test by making the number of seedlings of each test section into 20 pieces.

[Regeneration effect of pine root system by ectomycorrhizal fungi material]
1L of 1140-TK44 strain material or 1140 strain material prepared in Example 1 was applied within the root system range of black pine grown in advance, and the root system regeneration effect and mycorrhiza formation effect of the greening material were confirmed. Specifically, Japanese black pine is planted in the center of a planter with a capacity of 30L, cylindrical holes are drilled in two places on both sides, and the greening material is applied to these holes in 0.5L increments for cultivation. It was.

  Also, black pine (control group 1) to which compost was applied instead of greening material and black pine (control group 2) to which an equal mixture of red crust and river sand was applied were also cultivated in the same manner.

After continuing the cultivation for 6 months, the greening material, compost, or a mixture of red jade soil and river sand was dug from the soil, and the fine roots of black pine contained therein were washed out and dried, and the dry weight was measured. As a result, in the section where 1140-TK44 strain material or 1140 strain material was applied, many fine roots invaded and proliferated in the material. In particular, in the section where the 1140-TK44 strain material, which is a material according to the present invention, is applied, as shown in FIG. 9 (a), it is 7 times or more compared to the control group 2 , and 3.5 times or more compared to the control group 1. The amount of fine roots was confirmed, and a higher root regeneration effect was observed than in the case where 1140 strain material, which is a material of the comparative example, was applied.

  Moreover, when the mycorrhizal formation rate in the black pine of each division was measured, as shown in FIG.9 (b), in the division which applied the greening material of 1140-TK44 strain material or 1140 strain material, compared with the control districts 1 and 2. Many mycorrhizas were formed. Furthermore, a higher mycorrhizal formation rate was confirmed in the section where the 1140-TK44 strain material, which is the material according to the present invention, was applied, compared to the section where the 1140 strain material, which was the material of the comparative example, was applied. In addition, said mycorrhiza formation rate counted the total root tip number in a fine root, and calculated the ratio of the location in which the root tip is ectomycorrhizal root, and made it mycorrhizal formation rate. That is, the mycorrhizal formation rate = (external mycorrhiza root number / total number of fine roots) × 100. 9 (a) and 9 (b) show the average values of the results obtained by repeating the above test twice with six black pine in each test group.

[Regeneration effect of Yoshino cherry root system with Arbuscular mycorrhizal fungi]
Applying 1 L of greening material of KANSO-19 strain material (Example) or OG-105 strain material (Comparative example) within the range of pre-grown Yoshino cherry roots and continuing cultivation for 6 months, root regeneration by the greening material The effect and the mycorrhiza formation effect were confirmed. In addition, the root regeneration effect and the mycorrhiza formation effect were also confirmed for Yoshino cherry (control group 1) applied with compost instead of the greening material and Yoshino cherry (control group 2) applied with an equal mixture of red crust and river sand. In addition, application of the above-mentioned greening material, compost, and a mixture of red ball soil and river sand and evaluation of the root regeneration effect were performed by the same method as in Example 8. However, the evaluation of the mycorrhiza formation effect, that is, the measurement of the mycorrhiza formation rate is “Working with mycorrhizas in forestry and agriculture” (1996, Australian Center for International Agriculture). It was carried out by the method described on pages 182 to 184 of Cultural Research (issued by Australian Center for International Agricultural Research).

  As a result, as shown in FIG. 10 (a), a higher root-system regeneration effect was observed in the section where the greening material of KANSO-19 strain material or OG-105 strain material was applied as compared to the control group 1 and the control group 2. It was. In particular, in the section where the KANSO-19 strain material, which is a material according to the present invention, was applied, the amount of fine roots having a diameter of 2 mm or less was about twice that of the control group 1 and about 6 times that of the control group 2. In addition, a higher root regeneration effect was observed than in the section where OG-105 strain material, which is a material of the comparative example, was applied. Moreover, as shown in FIG.10 (b), in the area | region which applied the KANSO-19 strain material which is the material which concerns on this invention, a mycorrhiza higher than the area which applied OG-105 strain material which is a material of a comparative example The formation rate was confirmed. 10 (a) and 10 (b) show average values of results obtained by repeating the above test twice with six Yoshino cherry trees in each test section.

Claims (9)

What is obtained by holding mycelia of ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi in a plant having a pH of 5.0 to 8.0 by adding an acidic liquid, The ectomycorrhizal fungi or the arbuscular mycorrhizal fungi are Iuchichichichitaketake 1140-TK44 strain (NITE AP-01658), Xymetidae Shakashimeji 273-TS49 strain (NITE AP-01659), and Glomus genus KANSO- A greening material characterized in that it is any one or a plurality of 19 strains.   The greening material according to claim 1, wherein the acidic liquid is one or more of acetic acid, citric acid, and vinegar.   The greening material according to claim 1 or 2, wherein a particle size of the plant carbide is 2 mm to 10 mm. The greening material according to any one of claims 1 to 3, further comprising a spore suspension of the ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi.   It is a greening material applied for the recovery of the vigor of black pine, wherein the plant carbide in the greening material according to any one of claims 1 to 4, having a particle size of 2 to 6 mm and a carbonization temperature of 400 ° C to 800 ° C. Greening material characterized by charcoal and ectomycorrhizal fungi retained.   It is a planting material applied for the recovery of the pine tree, and in the planting material according to any one of claims 1 to 4, the plant carbide has a particle size of 2 to 6 mm and a carbonization temperature of 400C to 600C. Greening material characterized by charcoal and ectomycorrhizal fungi retained.   It is a tree planting material applied for restoring the tree vigor of a tree of the genus Sakura, wherein the plant carbide in the planting material according to any one of claims 1 to 4 has a particle size of 2 to 6 mm and a carbonization temperature of 400 ° C to A greening material characterized by charcoal at 800 ° C and retaining arbuscular mycorrhizal fungi.   A method for producing a greening material according to any one of claims 1 to 7, wherein the acidic liquid and sand are added to and mixed with the plant carbide, and the mixture is subjected to high-pressure sterilization and then exogenous in advance. Characterized in that a host plant infected with mycorrhizal fungi and / or arbuscular mycorrhizal fungi is planted in the mixture, and the mycelium of the ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi is elongated in the mixture. To make greening materials.   It is a method of manufacturing the greening material in any one of Claims 1-7, Comprising: The nutrient required for growth of the acidic liquid and the ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi to the plant charcoal And then sterilizing by high-pressure sterilization and then inoculating the plant charcoal with ectomycorrhizal fungi and / or arbuscular mycorrhizal fungi previously cultured separately, and extending the mycelium To make greening materials.

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