KR100239153B1 - Method of preparing va mycorrhizae inoculant - Google Patents

Abstract

The present invention is a general formula (I) in the production of VA mycorrhizal inoculation by cultivating plants infected with VA mycorrhizal in the presence of a carrier for VA mycorrhizal inoculation.

{Wherein R ¹ is a hydrogen atom or a methyl group, R ² is an oxygen atom or a sulfur atom, R ³ is a 2-4 alkyl group, 1-methyl-2propyl group or an allyl group of carbon, n is 2-5, m is 0 Or 1 each of the present invention} to provide a method for producing VA mycorrhizal inoculum is added to the carrier for VA myococcal inoculation. According to the production method of the present invention, the mycelial growth of VA mycorrhizal fungi is faster, and the formation of spores is also promoted.

In addition, by applying a specific compound of formula (I) to the plant under cultivation, contamination of various germs is also prevented.

Therefore, it is a manufacturing method of VA mycorrhizal inoculation which can produce VA mycorrhizal agent with high spore density and excellent activity of VA mycorrhizal at low cost.

Description

Preparation method of V. mycorrhizal inoculation carrier

The present invention relates to a method for preparing VA mycorrhizae inoculation useful in fields such as agriculture and horticulture. The present invention relates to a method for preparing VA inoculum inoculum having high spore density and activity by promoting the growth and sporulation of VA mycorrhizal fungi, and preventing the disease caused by P. aeruginosa, infectious diseases, and mycobacterial diseases.

VA mycorrhizal fungi are known to improve plant growth and disease resistance by infecting and coexisting with various plants. (Agriculture and Horticulture, Vol. 62, No. 8, pp. 930-937, 1987, Plant Protection Vol. 42, Vol. 5 Ho, pp. 259-266, 1988)

However, it is not known to be practical as VA myococcal inoculation which can be used for artificially growing VA mycorrhiza and infecting plants.

Conventionally known as VA mycorrhizal inoculum is that the VA mycobacterial spores have been adhered to the carrier with clay powder or manufactured by inoculation using a porous carrier such as foamed clay, but the activity is not sufficient and at high cost. It was not practical.

By the way, known methods of VA inoculation are divided into the following two ways.

① After spores are separated and recovered, the spores are mixed with an adhesive such as carboxymethyl cellulose on a carrier such as vermiculite, attapulgite, or diatomaceous earth, and then localized (JP-A-1-165369). Method of using carbon as a carrier (Japanese Patent Laid-Open No. 3-1030124).

② Carrier having a porous structure such as soil (Japanese Patent Laid-Open Publication No. 3-58715, 3-76572) or foamed clay pumice (Japanese Patent Laid-Open Publication No. 60-237987, 55-118390), A method in which the mycorrhizal fungi are propagated by using a ion exchanger (Japanese Patent Laid-Open No. 63-87973) to symbiize plant roots and mycorrhizal fungi, and using the mycorrhizal fungi attached to these carriers as inoculum.

However, in the above method of ①, spores are damaged at the granulation process, or spores are dried at the granulation process. On the other hand, in the method of ② described above, when natural soil is used, contamination by pathogens becomes a problem.

In addition, the microparticle soil was used as a carrier for VA mycorrhizal fungi, and the plants were grown at the interface between the coarse grain soil and the depth direction (Japanese Patent Application Laid-Open No. 3-76572). When VA fungus is recovered and used, the microparticle soil is crushed during the manufacturing process (2-4 months) and clogging occurs.

Therefore, in this state, supply of oxygen is not fully performed and it cannot be said that it is preferable for the growth of VA mycorrhizal bacteria.

In addition, there is a high risk of contamination of various germs such as pathogens.

Again, as described above, there is a method of propagating VA mycorrhiza by using a porous cationic exchanger as a carrier and potatoes as a host plant, but this method is not practical because the host plant is limited to potatoes and the carrier itself is expensive. .

In order to solve such a problem, the present inventors have conducted a thorough research, and as a result, by spraying a specific compound during the cultivation of plants inoculated with the VA mycorrhizal fungi, it is possible to prevent the occurrence of diseases of plants such as pythinm disease and Downy mildew disease. In addition, since mycelial growth of VA mycorrhizal was promoted and spore formation was also promoted, it was found that a cheap and practical VA mycorrhizal inoculated carrier was obtained and completed the present invention based on this finding. In other words, the present invention in the production of VA mycorrhizal inoculation by cultivating the plants infected with VA mycorrhizal in the presence of a carrier for VA mycorrhizal inoculation, Formula (I)

[Wherein R ¹ is a hydrogen atom or a methyl group, R ² is an oxygen atom or a sulfur atom, R ³ is an alkyl group having 2-4 carbon atoms, 1-methyl-2propyl group or an allyl group, n is 2-5, m is 0 or 1 shows each.] A method for producing a VA mycorrhizal inoculum, which is added to a carrier for VA mycobacterium inoculation, is provided.

VA mycorrhizal bacteria It is one kind of conjugated bacteria existing in the soil, and it is known that the mycelium adheres to the roots of various plants to form mycorrhiza and the symbiosis of both.

There are various kinds of mycorrhizal fungi used in the present invention, for example, genus gigaspora, genus akaolospora, genus entrophospora, genus sclerocystics, Microorganisms belonging to the genus of the scartellospora, glomus (glomus).

More specifically, for example, gigaspora margarita, acaulospora laevis, entrophospora inzrequens, sclerocystics dussi, scafellos Fort gregaria (scuttellospora gregaria), glomus mosseae, glomus intraradiciea, glomus caledonium, glomus fasciculatum, etc. Can be mentioned.

As a method for collecting these VA mycorrhizal fungi, a method of collecting by means of a sieve from nature (problem 5 regarding Suzuki's VA mycorrhiza, Agricultural Horticulture Vol. 62, No. 3, pp. 28-33, 1987) or by centrifugation (Japanese Patent Laid-Open No. 63-309178) is known.

In addition, methods of forming spores by aseptically growing VA mycorrhizal fungi by the nutrient thin film culture method (Japanese Laid-Open Patent Publication No. 55-118390) or the method of using organ-cultured roots (Japanese Laid-Open Publication No. 62-49037) There is also.

In addition, when the spores thus obtained are separated from the hyphae and handled alone, the spores are desirably stored in a wet state because their activity tends to decrease when dried.

In addition, it is desirable to store at low temperatures for suppressing the germination of spores, maintaining activity and preventing bacterial contamination.

In the present invention, the host plant which coexists with the VA mycorrhizal fungi is not particularly limited as long as it is a plant that grows quickly and extends well and is susceptible to infection with the VA mycorrhizal fungi, for example, corn crab, sorghum, barley, grass, sudangrass (sudan grass). Legumes, such as rice plants, eggplants, tomatoes, bell peppers, peppers, peppers, beans, peas, green beans, peanuts, legumes such as alfalfa clover, marigold, sunflower, cineraria, chrysanthemum Asteraceae plants, rose family plants such as strawberries, and lily plants such as garlic and onion are preferable.

In addition to sowing or seedlings, these plants are sown, seeded, transplanted, cultivated, cultivated, and seeded by cuttings, grafts and bulbs.

The VA mycorrhizal inoculation carrier (also referred to as a base material) of the plant used in the present invention is not particularly limited as long as it is inorganic and contains water and is difficult to disintegrate. It is preferable that the equipment including the treatment).

Specifically, calcined attapulgite calcined montmorillonite, calcined diatomaceous earth, zeolite, calcined pumpkin clay, pumice and the like are particularly preferable.

You may use plural different materials.

The calcined attapulgite and calcined montmorillonite are those having a particle diameter of 0.5-5 mm, preferably 1-3 mm, treated at a firing temperature of 200-1300 ° C., but granulated using a binder such as alumina as necessary. Used.

The calcined diatomaceous earth is treated with a particle diameter of 0.5-5 mm, preferably 1-3 mm, at a firing temperature of 200-1300 ° C., preferably 300-1000 ° C., but montmorillonite and etherpulzite clay may be used as a binder. There is a number.

Alumina or the like may also be used as the binder, but in this case, it is preferable to adjust the pH to 5.5-7.5.

Again, the zeolite preferably has a particle diameter of 0.5-5 mm, preferably 1-3 mm, not spherical in shape, and having a particle planar structure.

As the calcined amber clay, one having a particle diameter of 0.5-5 mm, preferably 1-3 mm, which has been treated at a firing temperature of 20-1300 ° C, preferably 200-1000 ° C is used.

In addition, the mixture of pumice in 1 part ratio with respect to 1-10 parts of said materials can be used suitably.

The particle diameter of pumice in this case is preferably 0.5-5 mm. The method of infection of VA mycorrhiza to host plants can be described as the application time of VA mycorrhizal fungi at any time before and after rooting of the host plant, but especially during the pre-treatment of sowing or cutting buds, at the same time as the seeding or cutting buds, or when transplanting seedlings. This is preferred.

In addition, it is preferable to apply the mixture to the equipment, to apply the layer to the lower layer of the seed or shoot, or to apply to the planting hole during planting. In addition, the amount of VA mycorrhizal fungi is not limited, but is usually about 1-100,000 per plant.

The infection of VA mycorrhiza to the host plant and the cultivation of the host plant may be carried out by a known method, for example, the temperature is 5-60 ° C, preferably 10-45 ° C, and the pH of the soil is 3-9.5, preferably It is performed on the conditions of 4-7.5.

When the host plant is cultivated in this way, infection with the VA mycorrhizal is established with the growth of the plant, and the VA mycobacterium grows.

In the present invention, in the cultivation of a plant infected with VA mycorrhizal in the presence of a carrier (material), the compound represented by the following general formula (I) is sprayed onto the VA mycorrhizal inoculation carrier.

[Wherein R ¹ is a hydrogen atom or a methyl group, R ² is an oxygen atom or a sulfur atom, R ³ is an alkyl group having 2-4 carbon atoms, 1-methyl-2propyl group or an allyl group, n is 2-5, m is 0 or 1 is represented respectively.] As the alkyl group having 2 to 4 carbon atoms, an ethyl group, a propyl group, a butyl group, a t-butyl group, an iso-butyl group and the like can be given.

As such a compound, what is represented by the following formula etc. is mentioned. In the formula, Me represents a methyl group, Et represents an ethyl group, Pr represents a propyl group, and Bu represents a butyl group.

There is no restriction | limiting in particular about the spreading method of the said compound, It becomes the form normally performed, for example, the plant is made into the aqueous solution of the density | concentration of 0.13-3.0 mg / L, Preferably it is 0.3-1.5 mg / L. Spray about 100-500ml per seedling.

In addition, in plant cultivation, water or nutrients can be supplied as needed.

Normally 2 to 5 months after the above-mentioned compound is sprayed, when the host plants are fully grown, the supply of water nutrients and the like is stopped and left for a while, the VA mycobacteria form spores.

VA mycorrhizal inoculation (formulation) is obtained by recovering the equipment with VA mycorrhizal spores formed here.

The VA mycorrhizal inoculation thus obtained is applied to a plant which can be infected with the VA mycorrhizal fungus and used for cultivation of various plants.

EXAMPLE

Next, the Example of this invention is demonstrated in detail.

[Example 1-3 and Comparative Examples 1,2]

15 pollen No. 6 (187 mm in diameter x 146 mm in height) were passed through a sieve of 3 mm mesh, and the remaining calcined montmorillonite was filled in a 1 mm sieve.

In each pot, 190 spores of the mycorrhizal fungus (Glomus faciculum) were placed on toilet paper at a depth of 3 mm so that the spores did not flow down with water.

Two seeds of corn (Goldenten DK649 from Kaneko Seed) were placed 1cm above.

Seeds of montmorillonite, VA mycorrhizal and corn in pollen were sufficiently wetted and then covered with vinyl sheet.

This was cultivated so that the water did not dry for 1 week while maintaining the inside of the glass greenhouse at 20-25 ℃.

Among the tombs that were grown, they were left with sound tombs, other seedlings were removed, and then grown in the same shape and watered daily in glass greenhouses.

One month after sowing, 300 ml of an aqueous solution (hereinafter abbreviated as medicament) prepared with propyl-3- (dimethyl amino) propyl carbamate hydrochloride (700 mg / l) was sprayed on five of the 15 pots. (Example 1), 10 other pots were watered only.

Then, once a week, 1000 times of Peter's liquid fertilizer (20-10-20) was sprayed and grown for 1 month.

At this point, the calcined montmorillonite at a distance of 5 cm from the root of the plant was removed as a sample by a hole, and the number of calcined montmorillonites to which the VA mycelium was attached was measured.

The average value per five flower pots is shown in the 1st table | surface with the number of VA mycorrhizal bacteria attached to total calcined montmorillonite allowance as a mycelial adhesion rate. As a control example, five of the ten pots without spraying the drug were randomly extracted, and the samples were taken out in the same shape to obtain the hyphae attachment rate of the five pollen sugars (Comparative Example 1). Once a week, the cultivation continued for a month, after which the spraying of liquid liquid fertilizer was stopped.

At that time, 300 ml of the same concentration was sprayed on the five pots sprayed with the medicine (Example 2), and another new pollen (not sprayed with the medicine in one month of cultivation) was sprayed with the same concentration on the five pots. Sprayed 300 ml each (Example 3). As a control, the remaining five pots were not sprayed with a medicament. (Comparative Example 2)

After stopping the supply of water liquid fertilizer and leaving it for 20 days, the pots were turned upside down and the calcined montmorillonite containing corn root and VA mycorrhizal was spread on the vinyl sheet.

The coarse roots of corn were removed and dried in the dark at 15 ° C. intact.

The sample thus obtained was randomly sampled from three places by 1 g per flower pot and the average value of the number of spores attached was measured.

The average value of the values obtained by the five flower pots of each example is shown in a 1st table | surface as VA mycorrhizal spore count.

[Examples 4 and 5 Comparative Examples 3 and 4]

The use of calcined aperitite in place of calcined montmorillonite as the carrier for inoculation and the use of glomus caldonium (Example 4) or glomus Mosesae (Example 5) in place of the glomus facilitum Others repeated the same operation as Example 2.

The number of spores adhered in the same manner as in Example 2 was measured. The average number is shown in a 2nd table | surface.

The same operation as in Example 4 was repeated except that no drug was sprayed (Comparative Example 3).

The same operation as in Example 5 was repeated except that no chemicals were sprayed (Comparative Example 4).

The number of spores adhered in the same manner as in Example 2 was measured.

The average number is shown in a 2nd table | surface.

Applicants are willing to provide samples of Glomus Calidonium or Glomus Mosesae at the request of any party.

According to the method of the present invention, the mycelial growth of VA mycorrhizal fungi is faster, and the formation of spores is also promoted. In addition, microbial contamination is also prevented by spraying drugs. Therefore, the VA mycorrhizal fungi having high spore density and excellent activity can be produced at low cost.

Therefore, VA mycorrhizal agent obtained by the present invention is effectively used as a practical material.

Claims (4)

In the preparation of VA mycorrhizal inoculation by cultivating a plant infected with VA mycorrhizal in the presence of a VA mycorrhizal inoculation carrier, Formula (I) [Wherein R ¹ is a hydrogen atom or a methyl group, R ² is an oxygen atom or a sulfur atom, R ³ is an alkyl group having 2-4 carbon atoms, 1-methyl-2propyl group or an allyl group, n is 2-5, m is 0 or 1 shows each.] A method for producing a VA mycobacterial inoculum, wherein the compound represented by the above is added to a VA mycobacterium inoculation carrier. The VA mycorrhizal fungus of claim 1, wherein the VA mycorrhizal fungus is at least one selected from the group consisting of at least gigaspora, acaulospora, entrophosphora, sclerosistis, skatellospora, and glomus. Preparation of Inoculum. The method of claim 1, wherein the VA mycorrhizal inoculation carrier is at least one member selected from the group consisting of calcined attapulite, calcined montmorillonite, calcined diatomaceous earth, zeolite, calcined amber clay, and pumice. The method of claim 1, wherein the compound of formula (I) is propyl-3- (dimethylamino) propyl carbamate hydrochloride.