JPH05244933A - Production of carrier for inoculating vesicular arbuscular mycorrhizae - Google Patents

Abstract

PURPOSE:To obtain the subject carrier having high spore density and excellent activity and useful for agricultural and horticultural use, etc., in high efficiency while preventing the proliferation of unwanted bacteria by spraying an agent such as disinfectant to a carrier for the inoculation of vesicular arbuscular mycorrhizae (VA mycorrhizae) and culturing a plant carrying the VA mycorrhizae. CONSTITUTION:One or more kinds of agents selected from disinfectants such as captan and nematocides such as 1,3-dichloropropene is scattered to a carrier for inoculating a VA mycorrhizae such as Glomus mosseae at an application rate of preferably 0.5-2g/m<2>. A plant carrying the VA mycorrhizae is cultured in the presence of the carrier to obtain the objective carrier.

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 fungus inoculation carrier useful in fields such as agriculture and horticulture. When producing a root fungus inoculation carrier, a specific drug is applied to prevent the occurrence of diseases such as Pythium disease, epidemics, and downy mildew.
Moreover, the present invention relates to a method for producing a VA mycorrhizal fungus inoculation carrier that promotes the growth and sporulation of VA mycorrhizal fungi and has a high spore density and excellent activity.

[0002]

PRIOR ART AND PROBLEM TO BE SOLVED BY THE INVENTION It is known that VA mycorrhizal fungi (Vesicular Arbuscular Mycorrhizae) can infect various plants and coexist with them to promote the growth and disease resistance of the plants. ("Agriculture and Horticulture", Vol. 62, No. 8, pages 930-937, 19)
1987; "Plant epidemics", Volume 42, No. 5, 259-2
66, 1988).

However, heretofore, practically no VA mycorrhizal inoculum that can be used to artificially grow VA mycorrhizal fungi and infect plants has been known. Conventionally known as a VA mycorrhizal inoculum is to make an inoculum using a separated VA mycorrhizal spores adhered to a carrier with clay powder or a porous carrier such as foamed clay. However, the activity was not sufficient, and it was expensive and not practical.

By the way, known methods for preparing a VA mycorrhizal inoculum can be divided into the following two methods. Separation and recovery of spores, vermiculite,
A method of granulating by mixing with a carrier such as diatomaceous earth together with an adhesive such as carboxymethyl cellulose (JP-A-1-16
5369) and a method of using charcoal as a carrier (JP-A-3-103124). Soil as a carrier (Japanese Patent Laid-Open No. 3-58715, 3)
-76572), foamed clay, pumice and the like having a porous structure (Japanese Patent Laid-Open No. 60-237987,
Japanese Patent Application Laid-Open No. 55-118390) and an ion exchanger (Japanese Patent Application Laid-Open No. 63-87973), and plant roots and VA
A method in which mycorrhizal fungi are symbiotic to grow VA mycorrhizal fungi, and the VA mycorrhizal fungi attached to these carriers are used as they are as an inoculum.

However, in the above method, the spores are damaged in the process of separating the spores and the granulation process, and the spores are killed at a high frequency due to the forced drying in the granulation process. Difficult to create. On the other hand, in the above method, when natural soil is used, contamination by pathogenic bacteria becomes a problem.

Further, as described above, there is a method of increasing VA mycorrhizal fungi by using a porous amphoteric ion exchanger as a carrier and using potatoes as a host plant, but this method is limited to host plants. Therefore, the carrier itself is expensive, which is not practical.

[0007]

[Means for Solving the Problems] As a result of intensive studies conducted by the present inventors in order to solve such problems, unexpectedly, during the cultivation of plants inoculated with VA mycorrhizal fungi, drugs were By spraying, it is possible to prevent the growth of miscellaneous bacteria, promote the growth of mycelia of VA mycorrhizal fungi, and also promote the formation of spores, so that an inexpensive and practical VA mycorrhizal inoculum carrier can be obtained. Based on this finding, the present invention has been completed.

That is, in the present invention, when a VA mycorrhizal fungal infected plant is cultivated in the presence of a VA mycorrhizal inoculum carrier to produce a VA mycorrhizal inoculum carrier, a bactericide and a nematicide are selected. Another object of the present invention is to provide a method for producing a carrier for inoculating VA mycorrhizal fungus, which comprises spraying at least one drug onto a carrier for inoculating VA mycorrhizal fungi.

VA mycorrhizal fungus is a kind of zygomycete existing in soil, and it is known that its hyphae form mycorrhizal roots of various plant roots and both of them coexist. There are various VA mycorrhizal fungi used in the present invention,
For example, the genus Glomus, Gigaspo
ra genus, Scutellospora genus, Acaulospora genus, Entrofospora
ntrophospora), Sclerocysti
s) Microorganisms belonging to the genus and the like. Among them, the genus Glomus, Gigaspora
Microorganisms belonging to the genus Scutellospora are preferred.

More specifically, for example, Glomus mosseae, Glomus intraradicies, Glomus caledonium, Glomus fasciculatum, and Gigaspora margarita. Gigaspora margarita), Scatellospora
Scutellospora gregaria, Acaulospora laevis, Entrophosporainfrequens
), Sclerocystis dus
si) and the like. Among these, Glomus mosseae, Glomus intraradicies, Glomus caledonium, Glomus fasciculatum,
Gigaspora margarita, Scutellospora gregaria
) Is preferred.

As a method for collecting these VA strains,
Method of collecting from nature using a sieve (Tatsuhiko Suzuki, Problems on VA mycorrhiza 5, Agriculture and Horticulture, Vol. 62, No. 3
No. pp. 28-33, 1987) and a method using centrifugation (Japanese Patent Laid-Open No. 63-309178).
Moreover, a nutrient thin film culture method (Japanese Patent Laid-Open No. 55-118390) or a method using organ-cultured roots (Japanese Patent Publication No. 62-4).
9037) and the like to spore formation by aseptically growing VA strains.

The host plant that is symbiotic with VA mycorrhizal fungi in the present invention is not particularly limited as long as it is a plant that grows fast and has well-rooted roots, and that VA mycorrhizal fungi are susceptible to infection. , For example, corn, mexico, sorghum (also known as sorghum or sorghum), wheat, turfgrass, grasses such as Sudangrass, eggplant, tomato, bell pepper, citrus and other solanaceous plants, soybeans, crow peas, mung beans, peanuts, alfalfa Beans such as clover, marigold, sunflower, cineraria, chrysanthemums such as chrysanthemum, roses such as strawberry, and lilies such as onion and onion are preferable. These plants are used not only as seeds and seedlings, but also after seeding and raising seedlings, and then transplanted and cultivated, vegetatively propagated, and grown and cultivated by cuttings, cuttings, grafts, bulbs, etc. .

The carrier for inoculating VA mycorrhizal fungi of a plant (also referred to as a base material) used in the present invention is not particularly limited as long as it is an inorganic substance and does not easily collapse when it contains water. However, from the viewpoint of preventing contamination of indigenous bacteria, a sterilized (including baked) base material is preferable. Specifically, calcined diatomaceous earth, zeolite, calcined red tama soil, pumice stone and the like are particularly preferable. Also, a plurality of different base materials may be used.

Here, the calcined diatomaceous earth has a particle size of 0.5 to 5 m.
m, preferably 1-3 mm, baking temperature 200-1300
What is treated at ℃, preferably 300-1000 ℃ is used. Alumina or the like can also be used as a binder, in which case it is preferable to adjust the pH to 5.5 to 7.5.

Further, the zeolite has a particle size of 0.5.
It is -5 mm, preferably 1-3 mm, and it is preferable that the surface is spherical and not smooth, and has a grain plane structure. As the calcined red dough, one having a particle size of 0.5 to 5 mm, preferably 1 to 3 mm, a calcining temperature of 200 to 1300 ° C., preferably
The one treated at 200 to 1000 ° C is used.

A method for infecting a host plant with VA mycorrhizal fungi will be described. The application time of VA mycorrhizal fungus may be either before or after rooting of the host plant, but especially for seeding and cutting. At the time of pretreatment, at the same time as sowing and cutting, or at the time of transplanting seedlings, it is preferable. In addition, as an application method, it is preferable to mix with a base material, apply in a layered form to a lower layer of seeds or sprouts, or apply in a planting hole at the time of planting.
Further, the application amount of VA mycorrhizal fungi is not particularly limited, but is usually about 1 to 100,000 per plant.

Infection of the host plant with VA mycorrhizal fungus and cultivation of the host plant may be carried out by known methods. For example, the temperature is 5 to 60 ° C., preferably 10 to 45 ° C., and the pH of the soil is 3 to 9. 0.5, preferably 4-7.5. When the host plant is cultivated in this way, the VA mycorrhizal fungus is infected and the VA mycorrhizal fungus grows as the plant grows.

In the present invention, when a plant infected with VA mycorrhizal fungus is cultivated in the presence of the above-mentioned base material, the drug is VA.
Spray on a carrier for inoculating mycorrhizal fungi. The drug used in the present invention is
1 or 2 selected from the group consisting of fungicides and nematicides
It means more than seeds. Specific examples of such an agent include, for example, a bactericide such as captan (captan).
) (Also known as orthoside), Benomyl (also known as Benlate), Ridomil (also known as Metalaxyl), Chloroneb
(Also known as Demosan), Ethazole (also known as Pansoil),
Thiabendazole, Oxamil (also known as dating)
And so on. Also, as nematicides, 1,3
-Dichloropropene (also known as Teron, DD) and the like.

The dose of such a drug is 0.1 to 10 g / m ² , preferably 0.5 to 2 g / m ² for the above drugs, and 0.01 to 10 g / m ^{2 for} the above drugs. , Preferably 0.05
~ 4 g / m ² . In the case of the drug of 0.05 to 10 g / m
² , preferably 0.1 to 2 g / m ² , in the case of
It is 0.005 to 12 g / m ² , preferably 0.01 to 3 g / m ² . Then, in the case of the drug of 0.005 to 6 g / m ² , preferably 0.01
To 1 g / m ^2, in the case of drug 0.1 to 10 g / m ^2, preferably 0.2 ~3 g / m ^2. Furthermore, in the case of the drug of 0.5 to 70 g / m ² , preferably 1 to 50 g / m ² ,
0.1 to 40 g / m ² , preferably 0.1 to 10
It is g / m ² . The above agents may be used alone or in combination of two or more kinds. Thus, by using various agents in combination, it is possible to prevent contamination with soil pathogens such as Fusarium, Pythium, Rhizoctonia, and Phytophytra, and nematodes such as mist nematodes and root-knot nematodes.

The treatment of the drug is carried out after the VA mycorrhizal fungus infects the plant. Usually, 7 days or more have passed since the application of VA mycorrhizal fungi to infect plants, preferably 1
The treatment of the drug is performed after the lapse of 0 days or more, more preferably 14 days or more and before the formation of spores. When two or more drugs are used in combination, they may be applied in a plurality of times at appropriate intervals.

There is no particular limitation on the method of spraying the above-mentioned drug, and any conventional method may be used.
1 to 3.0 mg / liter, preferably 0.3 to 1.5
As an aqueous solution having a concentration of mg / liter, about 100 to 500 ml per plant seedling may be sprayed in the vicinity of the seedling. In addition, when cultivating the plant, water can be supplied or nutrients can be supplied as necessary.

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

[0023]

EXAMPLES Next, the present invention will be described in more detail by way of examples.

Examples 1 to 10 and Comparative Examples 1 to 8 Black-black soil, river sand and red tama soil were mixed in equal amounts to prepare a substrate for culturing VA mycorrhizal fungi. This base material has a diameter of 2
4 × 23 cm deep (3.3 liters capacity) plastic pot up to a depth of 10 cm, on top of which VA mycorrhizal fungi [Glomus mosseae (Glomus mosseae) The spores were rejected at the site.)]] And 4,000 spores of each of them were sprayed, and the base material was placed on a thickness of 3 cm, and 25 Penn-Rings bentgrass seedlings (10 cm) were transplanted here. 1 in this pot
Eight of them were prepared and transferred to a greenhouse at 20 to 28 ° C. Fertilizer 1
Three liters of the following liquid fertilizer was given once a week. The surplus was discharged from the bottom of the pot. Liquid fertilizer was given ad libitum when the substrate was dry.

That is, as the liquid fertilizer, the following trace metal nutrient liquid and nutrient liquid fertilizer are used.
It was diluted 50 times, and 10 g of nutrient solution fertilizer was mixed with 1 liter of this diluted solution, and 0.25 g of magnesium sulfate heptahydrate was further added to prepare. The composition of the trace metal nutrients solution _{Fe-EDTA 0.12 H 2 BO 3} 2.86 MnCl 2 · 4 H 2 O 0.18 ZnSO 4 · 7 H 2 O 0.22 CuSO 4 · 5 H 2 O 0.08 Na 2 MoO 4 · 2 H 2 O 0.027 CoSO _{4 · 6 H 2 O 0.053 CaCl} 2 1.0 Al 2 (SO 4) 3 0.10 KI 0.10 KBr 0.10 10 liters of water nutrient liquid fertilizer compositions normally liquid fertilizer (N: P: K = 15 : 6: 6)

After transplantation, as shown in Table 1, given liters per pot were given and treated with the prescribed drug on the days corresponding to the prescribed number of days. 100 days after transplantation, fertilization, irrigation, and chemical treatment were completely stopped, and further 3 hours in the greenhouse.
It was left for 0 days. Then, the base material and the grown plant were taken out from the pot, the base material and the plant were separated with a sieve having an opening of 5 mm, and the base material was dried by ventilation at 25 ° C. The number of spores, the number of fusarium, and the number of pisium in 1 ml of this substrate were measured. The results are shown in Table 1.

In the Examples and Comparative Examples, the infection rate of VA mycorrhizal fungus when the drug was first added was measured, and the number of days after transplantation was 2 days, 6 days, 10 days
On the 20th and 40th days, 0%, 0%, 10%,
It was 15% and 20%. Here, the infection rate was determined by the following method. [Method of measuring infection rate] Trypan blue was used for staining according to the following procedure. (A) The infected plant root was washed thoroughly with water. (A) The drained roots were placed in a beaker and boiled with 10% KOH for 1 hour while slightly boiling. (C) If the root is colored, remove 10% KOH by rinsing it with water, and then dilute it with 10 times the hydrogen peroxide (about 3
%) Solution for 5 minutes and then washed with water. (D) Further, roots were dipped in a lactophenol-trypan blue dyeing solution having the composition shown below, and boiled gently for 5 to 30 minutes to dye. * Composition of lactophenol-trypan blue dyeing solution Carbolic acid (phenol) 200 ml Distilled water 200 ml Lactic acid 200 ml Glycerin 400 ml Trypan blue 1 g Total 1000 ml (e) Roots were washed with water and observed with a stereomicroscope. Infection rate calculation method The sample stained as described above was placed on a grid at 1 cm intervals, and a 10
Among 0 spots, the stained spots were counted and used as the infection rate.

[0028]

[Table 1]

[Footnote in Table 1] A: Captan (bactericide), application rate 0.25 g / pot B: Ethazole (bactericide), application rate 0.025 g / pot C: Thiabendazole (bactericide), application rate 0.15 g / Pot D: Teron (nematicide), application rate 0.5 g / pot E: Benlate (fungicidal agent), application rate 0.1 g / pot F: Lydomyl (fungicidal agent), application rate 0.1 g / pot G: Chloroneb ( Fungicide), application rate 0.2 g / pot

[0030]

INDUSTRIAL APPLICABILITY According to the method of the present invention, it is possible not only to effectively prevent the growth of germs such as pathogens and nematodes by spraying the drug during the cultivation of plants inoculated with VA mycorrhizal fungi. , VA Mycorrhizal mycelium growth is significantly promoted. Therefore, the formation of VA mycorrhizal spores is also promoted, and a VA mycorrhizal preparation having a high spore density and excellent activity can be manufactured at low cost. Therefore, the VA mycorrhizal preparation obtained by the present invention is effectively used as a practical material.

Claims (1)

[Claims] 1. A VA mycorrhizal fungus-infected plant is cultivated in the presence of a VA mycorrhizal fungus inoculating carrier to produce a VA mycorrhizal fungal inoculating carrier, wherein at least one selected from fungicides and nematicides is used. A method for producing a VA mycorrhizal fungus inoculating carrier, which comprises applying a seed medicine to a VA mycorrhizal fungal inoculating carrier.