JP3142675B2 - Method for producing carrier for inoculating VA mycorrhizal fungi - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a VA mycorrhizal inoculum-inoculated carrier useful in fields such as agriculture and horticulture. In manufacturing the mycorrhizal inoculum carrier, specific compounds are applied to prevent the disease such as picium disease, plague, and downy mildew, and to promote the growth and sporulation of VA mycorrhizal fungi to reduce the spore density. The present invention relates to a method for producing a highly inactive carrier of VA mycorrhizal fungi having excellent activity.

[0002]

2. Description of the Related Art It is known that VA mycorrhizal fungi (Vesicular Arbuscular Mycorrhizae) infect various plants and coexist with them, thereby promoting the growth of the plants and improving the disease resistance. (Agriculture and Horticulture, Vol. 62, No. 8, pp. 930-937, 19
1987; Plant Protection, Vol. 42, No. 5, 259-2
66, 1988).

[0003] However, V-mycorrhizal fungi that have been used to artificially grow and infect plants can be used.
Very few practical mycorrhizal inoculants are known. Conventionally, what is known as a VA mycorrhizal inoculant is:
Separated VA mycorrhizal fungal spores are bonded to a carrier together with clay powder, and inocula are prepared using a porous carrier such as foamed clay, but the activity is not sufficient, and it is expensive and practical. I could not say that.

[0004] By the way, known methods for preparing VA inoculum are divided into the following two methods. A method of separating and collecting spores, mixing the spores with a carrier such as vermiculite, attapulgite, and diatomaceous earth together with an adhesive such as carboxymethylcellulose and granulating the spores (Japanese Unexamined Patent Publication No. 1-16536).
No. 9) or a method using charcoal as a carrier (Japanese Unexamined Patent Publication No.
03124). Soil as carrier (Japanese Unexamined Patent Publication No.
No. 8715, No. 3-76572) and foamed clay, pumice, etc., having a porous structure (Japanese Patent Application Laid-Open No. 60-1985).
JP-A-237987, JP-A-55-118390), ion exchanger (JP-A-63-87973)
A method wherein the plant roots and the VA mycorrhizal fungi coexist to grow the VA mycorrhizal fungi, and the VA mycorrhizal fungi attached to these carriers are used as inoculants as they are.

[0005] However, in the above method, spores are damaged during the process of separating and granulating spores, and spores are frequently killed due to forced drying during the granulation process. It is difficult to create. On the other hand, in the above method, when natural soil is used, there is a problem of contamination by pathogenic bacteria. Further, in a method of cultivating a plant by filling the fine-grained soil as a carrier for VA mycorrhizal fungi and filling the interface in the depth direction with the coarse-grained soil (Japanese Unexamined Patent Publication No. Hei 3-76572), the method is described as follows.
The mycorrhizal fungi grown on 1 mm to 1 mm fine soil are collected and used. However, the fine soil is broken in the manufacturing process (2 to 4 months) and clogging occurs. Therefore, in this state, oxygen is not sufficiently supplied, which is not preferable for the growth of VA mycorrhizal fungi. In addition, there is a high risk of contamination by various bacteria such as pathogenic bacteria.

Further, as described above, there is a method of increasing VA mycorrhizal fungi using a porous amphoteric ion exchanger as a carrier and using potatoes as a host plant. However, this method is limited to a potato as a host plant. Is not practical because the carrier itself is expensive.

[0007]

Means for Solving the Problems In order to solve such problems, the present inventors have intensively studied and found that a specific compound was sprayed during cultivation of a plant inoculated with VA mycorrhizal fungi. Thereby, in addition to preventing the occurrence of plant diseases such as picium disease and downy mildew, the growth of hyphae of VA mycorrhizal fungi is promoted,
Further, since sporulation is also promoted, it has been found that an inexpensive and practical carrier for inoculating VA mycorrhizal fungi can be obtained, and the present invention has been completed based on this finding.

Namely, the present invention is the presence of VA mycorrhizae inoculation carrier, in producing the VA mycorrhizae inoculation carrier cultivates VA mycorrhizae infected plants, the general formula (I)

Embedded image Wherein R ¹ represents a hydrogen atom or a methyl group ;
Or 1. The method for producing a VA mycorrhizal inoculum inoculating carrier comprising spraying the compound represented by the formula

[0009] VA mycorrhizal fungi are a type of zygomycetes present in soil, and it is known that their mycelia form mycorrhizas in the roots of various plants, and both coexist. There are various types of VA mycorrhizal fungi used in the present invention,
For example, genus Gigaspora, genus Acaulospora, Entrophos
pora), Sclerocystis,
Genus Scutellospora, Glomus
mus).

[0010] More specifically, for example, Gigaspora margarita, Acaulospora laevis, Entrophospora infrequens, Sclerocystis dussi,
Scutellospora gregari
a), Glomus mosseae, Glomus intraradicie
s), Glomus caledonium
), Glomus fascic
ulatum).

[0011] As a method for collecting these VA strains,
A method of collecting from nature using a sieve (Tatsuhiko Suzuki, Problems on VA Mycorrhiza 5, Agriculture and Horticulture, Vol. 62, No. 3
Pp. 28-33, 1987) and a method by centrifugation (JP-A-63-309178).
In addition, a nutrient thin film culture method (Japanese Patent Application Laid-Open No. 55-118390) or a method using an organ-cultured root (Japanese Patent Publication No. Sho 62-4)
No. 9037), a method of aseptically growing a VA inoculum to form spores. When the spores thus obtained are separated from the mycelium and handled alone, it is preferable to store the spores in a wet state because the activity tends to decrease when dried. It is also desirable to store the spores at a low temperature in order to suppress germination of the spores, maintain the activity and prevent contamination of various bacteria.

The host plant that coexists with the VA mycorrhizal fungi in the present invention is not particularly limited as long as it is a plant that grows quickly and has a well-rooted plant and that is easily susceptible to VA mycorrhizal fungi. For example, grasses such as corn, crabgrass, sorghum (also known as sorghum or sorghum), wheat, turfgrass, and sudan grass; solanaceous plants such as eggplant, tomato, pepper, and shishito; soybeans, crow peas, mung beans, peanuts, alfalfa, Preferred are legumes such as clover, asteraceae such as marigold, sunflower, cineraria and chrysanthemum, roses such as strawberry, and lilies such as leek and onion. These plants are used as seeds and seedlings, or after sowing and raising seedlings, transplanted and cultivated, vegetatively propagated, or propagated and cultivated by cuttings, cuttings, grafts, bulbs, etc. .

As a carrier (also referred to as a base material) for inoculating a VA mycorrhizal fungus of a plant used in the present invention, any carrier can be used without particular limitation as long as it is inorganic and hardly disintegrated by containing water. From the viewpoint of preventing invasion of indigenous germs, a substrate subjected to a sterilization treatment (including a baking treatment) is preferable.
Specifically, calcined attapulgite, calcined montmorillonite, calcined diatomaceous earth, zeolite, calcined akadama clay, pumice stone, and the like are particularly preferable. Further, a plurality of different base materials may be used.

The calcined attapulgite or calcined montmorillonite has a particle size of 0.5 to 5 mm, preferably 1 to 5 mm.
３3 mm sintering temperature 200-1300 ° C, preferably 300
What has been treated at ~ 1000 ° C is used, and if necessary, granulated using a binder such as alumina is also used.

The calcined diatomaceous earth has a particle size of 0.5 to 5 mm,
Preferably, those having a size of 1 to 3 mm treated at a firing temperature of 200 to 1300 ° C., preferably 300 to 1000 ° C. are used.
Montmorillonite, attapulgite clay can be used as a binder. Alumina can also be used as a binder. In this case, the pH is preferably adjusted to 5.5 to 7.5.

Further, the zeolite has a particle size of 0.5 to
5 mm, preferably 1 to 3 mm, and those having a spherical surface and a non-smooth grain plane structure are preferred. The calcined red clay has a particle size of 0.5 to 5 mm, preferably 1 to 3 mm.
mm for baking temperature 200-1300 ° C, preferably 200-10
Those treated at 00 ° C. are used. In addition, the above-mentioned base materials 1 to 10
A mixture obtained by mixing pumice at a ratio of one part to one part can be suitably used. The particle size of the pumice in this case is 0.5-5mm
Are preferred.

The method of infecting host plants with VA mycorrhizal fungi is described below. The VA mycorrhizal fungi may be applied either before or after rooting of the host plant. It is preferable to perform the pre-treatment, at the same time as seed sowing or cutting buds, or at the time of transplanting seedlings. As an application method, it is preferable to mix with a base material, apply in a layered manner to the lower layer of seeds and buds, or apply in a planting hole at the time of planting.
The application rate of the VA mycorrhizal fungi is not particularly limited, but is usually about 1 to 100,000 per plant.

The infection of the host plant with VA mycorrhizal fungi and the cultivation of the host plant may be carried out by known methods, for example, at a temperature of 5 to 60 ° C, preferably 10 to 45 ° C, and a soil pH of 3 to 3 ° C.
To 9.5, preferably 4 to 7.5. When the host plant is cultivated in this way, the infection of the VA mycorrhizal fungi is established as the plant grows, and the VA mycorrhizal fungi proliferate.

In the present invention, when a plant infected with VA mycorrhizal fungus is cultivated in the presence of the substrate, the following general
The compound represented by the formula (I) is sprayed on a carrier for inoculating VA mycorrhizal fungi.

[0020]

Embedded image

In the formula, R ¹ is a hydrogen atom or a methyl group
And m represents 0 or 1. Examples of such compounds include those represented by the following formula. In addition,
In the formula, Me represents a methyl group , and Pr represents a propyl group.
You.

[0022]

Embedded image

[0023]

Embedded image

The method of spraying the above compound is not particularly limited, and may be in a usual manner. For example, the compound may be applied in an amount of 0.1 to 3.0 mg / liter, preferably 0.3 to 3.0 mg / l.
As an aqueous solution having a concentration of 1.5 mg / liter, about 100 to 500 ml per plant seedling may be sprayed near the seedling. In cultivation of the plant, watering and nutrients can be supplied as necessary.

Usually, about 2 to 5 months have passed since the above compound was sprayed, and when the host plant had grown sufficiently,
When supply of water, nutrients, etc. is cut off and left for a while, VA mycorrhizal fungi form spores. Then, the substrate (formulation) inoculated with the VA mycorrhizal fungus can be obtained by collecting the substrate to which the formed VA mycorrhizal fungal spores adhere. The VA mycorrhizal fungus inoculation carrier thus obtained is applied to plants that can be infected with VA mycorrhizal fungi and used for cultivation of various plants.

[0026]

Next, the present invention will be described in detail with reference to examples. Examples 1 to 3 and Comparative Examples 1 and 2 15 flower pots No. 6 (187 φ × 146 mmH, hereinafter abbreviated as pots) were passed through a sieve having a mesh size of 3 mm to obtain 1 mm.
The baked montmorillonite remaining on the sieve was spread. In each pot, 190 spores of VA mycorrhizal fungus [G. fasciculatum (this fungus was rejected by the Institute of Microbial Engineering, National Institute of Advanced Industrial Science and Technology)] The spores were placed on tissue paper at a depth of 3 mm so that the spores did not flow down with the water. Two seeds of corn (variety: Golden Tent DK649: cat seedlings) were placed 1 cm above it.
The fired montmorillonite, VA mycorrhizal fungi and corn seeds in the pot were sufficiently wetted and covered with a vinyl sheet. This was cultivated for one week without water, while maintaining the glass greenhouse at 20 to 25 ° C.

[0027] Among the grown seedlings, other seedlings were removed except for healthy seedlings, and then cultivated in a glass greenhouse while being irrigated daily. One month after the seeding date, 300 ml of an aqueous solution (hereinafter abbreviated as drug) prepared to 700 mg / l of propyl-3- (dimethylamino) propyl carbamate hydrochloride was sprayed on 5 of the 15 pots. (Example 1). The other 10 pots received only water. Thereafter, once a week, Peters liquid fertilizer (20-10-2)
The solution was sprayed with a 1000-fold solution of 0) and cultivated for one month.

At this time, the calcined borer montmorillonite at a distance of 5 cm from the root of the plant was sampled by a cork borer, and the number of calcined montmorillonite to which the mycelium of VA was attached was counted. Table 1 shows the average value per 5 pots as the number of mycelia adhering to the number of adhering VA mycorrhizal fungi per the total number of calcined montmorillonite. In addition, as a control, 5 pots were randomly extracted from 10 pots not sprayed with the above-mentioned drug, samples were sampled in the same manner, and the hyphal adhesion rate per 5 pots was determined (Comparative Example 1).

Further, while applying the liquid fertilizer once a week,
The cultivation was continued for three months, after which the application of water and liquid fertilizer was stopped. At that time, 300 ml of the same concentration of the drug was sprayed on the five pots on which the drug was sprayed first (Example 2), and another new pot (the drug was not sprayed in one month of cultivation)
Five pots were sprayed with the same concentration of the drug at 300 ml each (Example 3). As a control, the remaining 5 pots were not sprayed with the drug (Comparative Example 2).

Thereafter, the supply of water and liquid fertilizer was stopped, and the pot was turned over, and the corn root and the VA were turned over.
The calcined montmorillonite containing mycorrhizal fungi was spread on a vinyl sheet. The thick root of corn was removed and dried as it was in a dark place at 15 ° C. The sample thus obtained was randomly sampled from three places at 1 g per pot, and the average value of the number of spores attached was measured. Table 1 shows the average of the values obtained in 5 pots in each section as the number of spores of mycorrhizal VA.

[0031]

[Table 1] Table 1

Examples 4-5 In Example 2, calcined attapulgite was used in place of calcined montmorillonite, and as VA mycorrhizal fungi,
Glomus caledonium (Example 4) or Glomus mosseae (Example 5) instead of Glomus facilitatorum
[Both bacteria were rejected at the Institute of Microbial Engineering, National Institute of Advanced Industrial Science and Technology. Example 2 was used except that
A corn seedling was cultivated in the same manner as described above, a sample was collected, and the average value of the number of attached spores was measured. Each ward 5
The average of the values obtained in the pot is shown in Table 2 as the number of spores of mycorrhizal fungi.

Comparative Examples 3 to 4 The same procedure as in Examples 4 to 5 (Comparative Example 3) or Example 5 (Comparative Example 4) was carried out except that no chemical was sprayed. The number of fungal spores was determined. The results are shown in Table 2.

[0034]

[Table 2] Table 2

[0035]

According to the method of the present invention, the growth of hyphae of VA mycorrhizal fungi is accelerated and the formation of spores is promoted. In addition, bacterial contamination is prevented by spraying the medicine. Therefore, the spore density of VA mycorrhizal fungi is high and V
A mycorrhizal fungal preparation can be produced at low cost. Therefore, the VA mycorrhizal fungal preparation obtained by the present invention is effectively used as a practical material.

──────────────────────────────────────────────────続 き Continuation of the front page (51) Int.Cl. ⁷ Identification code FIC12R 1: 645) (56) References JP-A-3-247270 (JP, A) JP-A-3-76572 (JP, A) JP-A-60-237987 (JP, A) JP-A-63-87973 (JP, A) JP-A-50-157152 (JP, A) JP-A-50-68835 (JP, A) (58) (Int.Cl. ⁷ , DB name) C12N 11/02 CA (STN)

Claims (1)

(57) [Claims] The presence of 1. A VA mycorrhizae inoculation carrier, in producing the VA mycorrhizae inoculation carrier cultivates VA mycorrhizae infected plants, the general formula (I) ## STR1 ## Wherein R ¹ represents a hydrogen atom or a methyl group ;
Or 1. A method for producing a carrier for inoculating VA mycorrhizal fungi, comprising spraying the compound represented by formula (I) on a carrier for inoculating VA mycorrhizal fungus.