JP2009196920A - Method for preserving microbial agrochemical formulation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for preserving a microbial agrochemical, which can be suitably used for maintaining the viable cell count in a microbial agrochemical formulation containing microbial cell bodies stably for a long period of time. <P>SOLUTION: The method uses a package containing a desiccating agent in drying a microbial agrochemical formulation composition. The microbial agrochemical formulation is more preferably a microbial agrochemical formulation containing dried microbial cell bodies produced by mixing (A) sodium chloride and/or potassium chloride and/or (B) trehalose or sucrose and freeze-drying the mixture. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a method for preserving a microbial pesticide preparation containing microbial cells.

  When commercializing microbial pesticides, it is necessary to be able to maintain a sufficient number of viable bacteria in the process of production, distribution and use. If the number of viable bacteria is greatly reduced before reaching the site of use, it will be difficult to exert its effect, which will be a problem in quality control. Therefore, a preservation technique that does not kill the cells as much as possible has been required.

  As a technique for preserving microorganisms, Patent Document 1 describes an immobilized product of Pseudomonas bacteria, a method for immobilizing the same, and a method for formulating an above-ground plant disease control agent comprising the immobilized product. This is to obtain an immobilized product with good viability recovery and storage stability by mixing trehalose with Pseudomonas bacteria, freezing and then vacuum drying.

Further, Patent Document 2 describes a microbial pesticide preparation obtained by adding a bag containing an adsorbent having ammonia adsorption ability to a microorganism having a control effect against plant diseases. Furthermore, Patent Document 3 describes a microbial pesticide preparation in which a microorganism having a controlling effect against plant diseases is freeze-dried and then mixed with an adsorbent having ammonia adsorption ability.
The inventions of Patent Document 2 and Patent Document 3 both use zeolite, molecular sieves, silica gel or the like as an adsorbent having ammonia adsorbing ability, and are microorganisms having an effect of controlling plant diseases, such as Erwinia bacterium or Pseudomonas genus. It makes it possible to store bacteria in a more stable state.

  Patent Document 4 describes a hydrating composition for agriculture that uses filamentous fungal spores as an active ingredient, can stably store the filamentous fungal spores for a long period of time, and has good physical properties during application. Hydrating compositions containing filamentous fungal spores having a specific particle size distribution and adsorbents having a specific amount of water-absorbing ability as essential components have good preservability and physical properties at the time of application. By enclosing it in a permeable bag-like body, it can be stored stably for a long period of time.

The inventors lyophilized the cells by the method of Patent Document 1 using Pseudomonas rhodesia 050572I9 strain. However, the number of viable cells in the lyophilized product of the cells decreased with the passage of time, and good results were not obtained as a long-term stable storage method.
In addition, even with the method described in Patent Document 4, Pseudomonas rhodesia does not form spores unlike filamentous fungi, and thus the method described does not provide sufficient long-term stability.
Moreover, in patent document 4, it is pointed out that the preservation | save method of patent document 2 and patent document 3 is inconvenient at the time of chemical | medical agent application, especially when applying a seed dressing process or using water.

JP 2005-325077 A JP 2000-264807 A JP 2000-264808 A JP 2004-123606 A

  An object of the present invention is to provide a method for preserving microbial pesticides, which makes it possible to stably maintain the number of viable bacteria in a microbial pesticide preparation containing microbial cells.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that a dried product obtained by freeze-drying a Pseudomonas rhodesia strain having control ability against plant diseases is a resin containing a desiccant in a high concentration. It has been found that the number of viable bacteria in the dried bacterial cell can be stably maintained for a long time by encapsulating with a packaging material based on the material, and the present invention has been achieved.

That is, the present invention is (1) a method for preserving a microbial pesticide preparation characterized by using a packaging material containing a desiccant,
(2) Microbial pesticide formulation
(A) It is a microbial pesticide preparation containing a dried microorganism cell product obtained by mixing sodium chloride and / or potassium chloride and / or (B) trehalose or sucrose and freeze-drying. , (1) storage method,
(3) The preservation method according to (1) or (2), wherein the microorganism used in the microbial pesticide preparation is Pseudomonas rhodesia having an ability to control plant diseases.

The present invention also provides:
(4) The preservation method of (3), wherein the Pseudomonas rhodesia having the ability to control plant diseases is Pseudomonas rhodesia 050572I9 strain (international accession number FERM BP-10912),
(5) The storage method according to (1) to (4), wherein the packaging material has a bag shape.

  According to the present invention, the number of viable bacteria in the microbial pesticide preparation composition can be stably maintained for a long time.

  The present invention is a method for preserving a microbial pesticide preparation, wherein the microbial pesticide preparation is encapsulated using a packaging material containing a desiccant.

(Wrapping material)
The packaging material used in the present invention is characterized by containing a desiccant. Although it does not specifically limit as a raw material of a packaging material, Resin, a metal, paper, those composite materials, etc. are mentioned, Resin is preferable. Examples of the resin include polyethylene, polyvinyl chloride, polyvinylidene chloride, polyvinyl alcohol, polypropylene, polyester, polycarbonate, polystyrene, polyacrylonitrile, ethylene vinyl acetate copolymer, ethylene-vinyl alcohol copolymer, ethylene-methacrylic acid copolymer. , Nylon, cellophane and the like.
Examples of the desiccant include zeolite, molecular sieve (trade name), silica gel, aluminum oxide, calcium chloride, calcium oxide, magnesium oxide, activated anhydrous calcium sulfate, magnesium sulfate, phosphorus oxide (V), potassium carbonate, potassium hydroxide, and the like. Can be mentioned.
In the present application, the packaging material containing a desiccant is not particularly limited as long as the desiccant is contained in the packaging material. For example, the desiccant may be kneaded into the base material of the packaging material, and the desiccant may be uniformly or non-uniformly dispersed in the packaging material. May be a layer of a desiccant or a layer containing a desiccant. The content of the desiccant in the packaging material is not particularly limited as long as the effects of the invention can be obtained, but the content is preferably high.
Specific examples of the resin containing a desiccant include a moisture-absorbing resin “Moist Catch (registered trademark)” (manufactured by Kyodo Printing Co., Ltd.) using a resin containing a high amount of synthetic zeolite as a raw material.

  In the present invention, by encapsulating the microbial pesticide preparation in the packaging material, it is possible to maintain a dry state for a long period of time and to stabilize the microbial pesticide preparation. Examples of the form of encapsulation include blister packaging, shrink packaging, caramel packaging, encapsulation in a bag, and the like, and encapsulation in a bag is preferable. Examples of the shape of the wrapping material include a bag shape, a film shape, a cylindrical shape, and the like, and preferably a sealable bag shape.

(Microbial pesticide formulation)
The present invention is a method for preserving a microbial pesticide preparation using the packaging material described above. The microorganism used in the microbial pesticide preparation is not particularly limited, but is more preferably Pseudomonas rhodesia having an ability to control plant diseases. About Pseudomonas rhodesia having the ability to control plant diseases, it is described in International Patent Application No. PCT / JP2007 / 71531 (filed on Nov. 6, 2007).

  Among the microorganisms used in the method of the present invention, among Pseudomonas rhodesiae, those having high ability to control plant diseases are more preferable. For example, Pseudomonas rhodesia 050572I9 strain (International Accession Number FERM BP-10912), Pseudomonas rhodesia JCM11940 strain and mutant strains of these strains are more preferable, and Pseudomonas rhodesia 050572I9 strain and mutant strains thereof are more preferable.

  These Pseudomonas rhodesias are scab, perforated bacterial disease, soft rot, spotted bacterial disease, black spot bacterial disease, bacterial wilt disease, brown spot bacterial disease, stem rot bacterial disease, rice blast bacterial disease, seedling bacterial disease It is a microorganism having an effect on plant diseases such as white leaf blight, rot, and black rot.

  In addition, the plant to which the disease control of the present invention is applied is not particularly limited as long as the microorganism of the present invention is a plant that can exert the control ability. For example, cruciferous, eggplant, cucurbitaceae, liliaceae, leguminous, asteraceae, redwood, rice, rose, radish, primrose, citrus, grape, matabidae, persimmonaceae, sericaceae, The plants belonging to the convolvulaceae family or the taro family can be mentioned. Among them, plants belonging to the Brassicaceae family such as Chinese cabbage, plants belonging to the Compositae family such as lettuce, plants belonging to the solanaceous family such as potato, mandarin oranges such as lemon and navel Preferred examples include plants belonging to the family, plants belonging to the rose family such as peaches, and the like.

The microbial pesticide preparation used in the present invention is a microbial pesticide preparation containing a dried microbial cell product obtained by freeze-drying a microorganism. For lyophilization of microorganisms, salts and / or saccharides can be added and lyophilized. Examples of salts include sodium chloride, potassium chloride, potassium citrate, sodium citrate and the like. Examples include glucose, lactose, sucrose, raffinose, sorbitol, xylitol, inositol, trehalose, palatinose, and palatinit.
Each of these salts and saccharides may be used alone or in combination of two or more. Among these, a microbial pesticide preparation containing a dried microorganism cell product obtained by mixing trehalose and / or sucrose and freeze-drying is preferable, and further sodium chloride and / or potassium chloride is mixed. More preferably, it is a microbial pesticide preparation containing a dried microbial cell product obtained by freeze-drying.

The sodium chloride and / or potassium chloride content is preferably added to the freeze-dried cell suspension to a concentration of 0.1% or more and less than 2.0%, alone or mixed.
By mixing sodium chloride and / or potassium chloride into the bacterial cell culture solution, the protective effect of the bacterial cells during lyophilization can be obtained. Furthermore, the effect of stabilizing the dried microbial cell can also be obtained.

The trehalose and / or sucrose content is preferably added to the lyophilized cell suspension so as to have a concentration of 10% or more and less than 50%.
By further mixing trehalose or sucrose with the bacterial cell culture solution, the protective effect of the bacterial cells during lyophilization can be obtained.

  The microbial pesticide preparation of the present invention may contain other optional components depending on the desired dosage form and effect. The optional component is not particularly limited as long as the effects of the present invention are not impaired, and examples thereof include a carrier, a surfactant, a dispersant, and an auxiliary agent. Further, if necessary, an antioxidant, a colorant, a lubricant, an ultraviolet ray inhibitor, an antistatic agent, a preservative, and the like can be added.

  Carriers include inorganic salts such as calcium carbonate, potassium chloride, sodium sulfate, calcium sulfate and ammonium sulfate; organic acids such as citric acid, malic acid and stearic acid; and salts thereof; sugars such as glucose, lactose and sucrose; alumina Solid carriers such as powder, silica gel, zeolite, hydroxyapatite, zirconium phosphate, titanium phosphate, titanium oxide, zinc oxide, hydrotalcite, kaolinite, montmorillonite, talc, clay, diatomaceous earth, bentonite, white carbon, kaolin, vermiculite Can be mentioned.

Further, the surfactant (which can also be used as a dispersant) is not particularly limited as long as it can be used in ordinary agricultural and horticultural preparations. Specifically, the following nonionic surfactants are used. Agents, anionic surfactants, cationic surfactants, and amphoteric surfactants.
Nonionic surfactants include sorbitan fatty acid esters (C12-18), POE sorbitan fatty acid esters (C12-18), sugar ester type surfactants such as sucrose fatty acid esters; POE fatty acid esters (C12-18), Fatty acid ester type surfactants such as POE resin acid ester and POE fatty acid diester (C12-18); Alcohol type surfactants such as POE alkyl ether (C12-18); POE alkyl (C8-12) phenyl ether, POE dialkyl (C8-12) alkyl ether type surfactants such as phenyl ether, POE alkyl (C8-12) phenyl ether formalin condensate; polyoxyethylene polyoxypropylene block polymer; alkyl (C12-18) polyoxyethylene poly Polyoxyethylene / polyoxypropylene block polymer type surfactants such as xylpropylene block polymer ether; alkylamine type surfactants such as POE alkylamine (C12-18) and POE fatty acid amide (C12-18); POE fatty acid bis Bisphenol type surfactants such as phenyl ether; polyaromatic surfactants such as POA benzylphenyl (or phenylphenyl) ether and POA styrylphenyl (or phenylphenyl) ether; POE ether and ester type silicon and fluorine-based surfactants Examples thereof include silicon-based and fluorine-based surfactants such as agents; vegetable oil-type surfactants such as POE castor oil and POE hydrogenated castor oil; and the like.

  Examples of the anionic surfactant include alkyl sulfate (C12-18, Na, NH4, alkanolamine), POE alkyl ether sulfate (C12-18, Na, NH4, alkanolamine), POE alkylphenyl ether sulfate (C12-18, NH4, alkanolamine), POE benzyl (or styryl) phenyl (or phenylphenyl) ether sulfate (Na, NH4, alkanolamine), polyoxyethylene, polyoxypropylene block polymer sulfate (Na, NH4, alkanolamine) and other sulfates Type surfactants: paraffin (alkane) sulfonate (C12-22, Na, Ca, alkanolamine), AOS (C14-16, Na, alkanolamine), Alkylsulfosuccinate (C8-12, Na, Ca, Mg), alkylbenzenesulfonate (C12, Na, Ca, Mg, NH4, alkylamine, alkanol, amine, cyclohexylamine), mono- or dialkyl (C3-6) naphthalenesulfonate (Na, NH4, alkanolamine, Ca, Mg), naphthalenesulfonate / formalin condensate (Na, NH4), alkyl (C8-12) diphenyl ether disulfonate (Na, NH4), lignin sulfonate (Na, Ca), POE alkyl (C8-12) sulfonate surfactants such as phenyl ether sulfonate (Na), POE alkyl (C12-18) ether sulfosuccinic acid half ester (Na); carboxylic acid type fatty acid salt (C12-18, N , K, NH4, alkanolamine), N-methyl-fatty acid sarcosinate (C12-18, Na), resinate (Na, K), etc. POE alkyl (C12-18) ether phosphate (Na, alkanolamine), POE mono Or dialkyl (C8-12) phenyl ether phosphate (Na, alkanolamine), POE benzyl (or styryl) phenyl (or phenylphenyl) ether phosphate (Na, alkanolamine), polyoxyethylene / polyoxypropylene block polymer (Na , Alkanolamine), phosphatidylcholine / phosphatidylethanolimine (lecithin), phosphate type surfactants such as alkyl (C8-12) phosphate; and the like.

  Cationic surfactants include alkyltrimethylammonium chloride (C12-18), methyl polyoxyethylene alkylammonium chloride (C12-18), alkyl N-methylpyridium bromide (C12-18), mono- or dialkyl. (C12-18) ammonium type surfactants such as methylated ammonium chloride and alkyl (C12-18) pentamethylpropylenediamine dichloride; alkyldimethylbenzalkonium chloride (C12-18), benzethonium chloride (octylphenoxyethoxyethyldimethylbenzyl) Benzalkonium type surfactants such as ammonium chloride).

Examples of amphoteric surfactants include betaine surfactants such as dialkyl (C8-12) diaminoethylbetaine and alkyl (C12-18) dimethylbenzylbetaine; dialkyl (C8-12) diaminoethylglycine, alkyl (C12-18) Glycine-type surfactants such as dimethylbenzylglycine;
Said surfactant and / or dispersing agent can be used individually by 1 type or in mixture of 2 or more types.

  Examples of the auxiliary agent include carboxymethyl cellulose, polyethylene glycol, gum arabic, and starch.

(Manufacture of microbial pesticide formulations)
The microbial pesticide preparation of the present invention is a microbial pesticide preparation containing a dried microbial cell product obtained by freeze-drying microorganisms, and the lyophilized product of microorganisms can be obtained by known apparatuses and conditions.
For example, when the microorganism is Pseudomonas rhodesia 050572I9 strain, the microorganism is shake-cultured in a liquid standard medium (yeast extract 0.25%, peptone 0.5%, glucose 0.1%, pH 7.0), and then centrifuged. Or by membrane filtration.
The obtained cell suspension is mixed with salts and / or sugars as necessary, and lyophilized. Freeze-drying can be performed by known apparatuses and conditions. By pulverizing after freeze-drying, a powdered dried microbial cell product is obtained.

By selecting and mixing arbitrary components according to a desired dosage form into the obtained dried microbial cell product, a microbial pesticide preparation can be obtained. The dosage form of the microbial pesticide preparation of the present invention is preferably a dosage form such as powder, wettable powder, granule, etc., more preferably wettable powder.
The form of the microbial pesticide preparation of the present invention is not particularly limited, and forms that can be taken by ordinary agricultural and horticultural pesticides, such as powders, wettable powders, emulsions, flowables, granules and the like, can be employed.

The obtained microbial pesticide preparation is immediately enclosed in a packaging material containing a desiccant after preparation. By using the packaging material, the microbial pesticide preparation can be stably stored for a long time. The packaging using the packaging material may be vacuum packaging or normal pressure packaging. In the packaging material, an agent other than the microbial pesticide preparation may be encapsulated. For example, a desiccant, a hygroscopic agent, a humidity control agent, or the like may be enclosed.
The temperature at which the microbial pesticide formulation enclosed in the packaging material is stored is not particularly limited, but it can be stored at room temperature, more preferably 10 ° C or lower, and 4 ° C (refrigerator) or lower. Is more preferable.

(Use of microbial pesticides)
The disease to which the microbial pesticide preparation of the present invention is applied is not particularly limited as long as it has an effect. For example, when the microorganism is Pseudomonas rhodesia that controls plant diseases, there is no particular limitation as long as the disease is a plant disease caused by infection of the plant with a pathogenic fungus that exhibits the ability to control Pseudomonas rhodesia. For example, scab disease, perforated bacterial disease, soft rot, spotted bacterial disease, black spot bacterial disease, bacterial wilt disease, brown spot bacterial disease, stem blight bacterial disease, rice wilt bacterial disease, seedling bacterial disease, white leaf blight , Rot, black rot, and the like.
In addition, plant diseases to which plant disease control agents are applied include scab, perforated bacterial disease, soft rot, spotted bacterial disease, black spot bacterial disease, bacterial wilt disease, brown spot bacterial disease, stem rot bacterial disease, One or more, preferably two or more, more preferably diseases of all kinds of plants selected from blast bacterial disease, seedling bacterial disease, white leaf blight, rot, and black rot. In particular, at least one of soft rot, rot, black rot, scab, and perforated bacterial disease, preferably two or more, and more preferably all diseases.

  The method for using the microbial pesticide preparation of the present invention is not particularly limited as long as it is a method for treating the microbial pesticide preparation of the present invention on a plant and / or the cultivated soil of the plant. Similarly to the above, it can be appropriately selected depending on the type of plant disease, the type of applied plant, and the like. For example, the plant disease control agent of the present invention may be treated on plants by directly applying or spraying the microbial pesticide formulation of the present invention to plants, or the plant is grown using the microbial pesticide formulation of the present invention. The microbial pesticide preparation of the present invention may be processed into plant cultivation soil by mixing, spraying or irrigating the soil (plant cultivation soil). Here, when the microbial pesticide preparation of the present invention is treated on the cultivated soil of the plant, the plant may be planted after treating the microbial pesticide preparation of the present invention on the soil, and after planting the plant on the soil, You may process to soil. Moreover, as described in JP-A-2001-302407, the microbial pesticide preparation of the present invention is installed in the vicinity of the air outlet of a blower that blows air in the facility, and the pesticide is sprayed together with the air sent from the air outlet. it can.

When the microbial pesticide preparation of the present invention is treated on a plant and / or cultivated soil of the plant, the microbial pesticide preparation of the present invention can be diluted with an appropriate amount of water or the like. The amount of the microbial pesticide preparation of the present invention to be treated on the plant and / or the cultivated soil of the plant cannot be defined unconditionally because it differs depending on the type of plant disease, the type of applied plant, etc. In terms of the cell concentration of Pseudomonas rhodesia, it can be usually in the range of 1 × 10 ² to 1 × 10 ¹¹ cfu / ml, preferably 1 × 10 ⁴ to 1 × 10 ⁹ cfu / ml.
In addition, the number of spraying times may be appropriately selected depending on the type of plant disease, the type of applied plant, the degree of disease, and the like.

EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.
In addition, the effect of the preservation | save method of the microbial pesticide preparation which is this invention can be confirmed by measuring the viable cell number in a microbial cell dry substance, and calculating the survival rate of a microbial cell.
[Method for measuring the number of viable bacteria] 1 g of the sample was weighed and made up to 10 mL with sterilized water. Thereafter, the number of colonies grown was appropriately diluted and grown by a plate dilution method, and the number of viable bacteria was determined. A standard agar medium (Nissui Pharmaceutical Co., Ltd.) was used and cultured at 25 ° C. overnight.

(Preparation of dried cells)
1. Preparation of cell suspension A 300 mL Erlenmeyer flask was charged with 150 mL of a standard liquid medium (yeast extract 0.25%, peptone 0.5%, glucose 0.1%, pH 7.0) and sterilized by heating. 0.1 mL of a preculture of Pseudomonas rhodesia 050572I9 strain (FERM BP-10912 strain) was inoculated and cultured in a reciprocating shaker at 30 ° C. and 100 rpm for 3 days. The obtained culture broth was centrifuged to remove the supernatant, and the precipitate was washed with water, and centrifuged again to remove the supernatant, and the precipitate was washed with water.
2. Drying of bacterial cells To the prepared bacterial cell suspension, NaCl (or KCl) and sucrose (or trehalose) are added to the concentrations shown in the table below, followed by lyophilization. Obtained. The concentration is the concentration in the cell suspension.
Moreover, the thing which does not add NaCl etc. was set as control.
In each Example, the number of viable cells in the dried cell body after freeze-drying relative to the number of viable cells in the cell suspension before lyophilization is shown in Table 1 as the survival rate.

As shown in the dried microbial cell 2, the survival rate was 28.7% just by adding NaCl. However, when NaCl was added in an amount of 2% or more, the survival rate significantly decreased.
As shown in the dried bacterial cells 4, 6, and 8, the survival rate became 80% or more by further adding sucrose.

[Example 1]
The dried microbial cell product 7 was enclosed in a bag of a desiccant-containing packaging material (“Moist Catch (trade name)”, manufactured by Kyodo Printing Co., Ltd.). As a comparative example, a dried microbial cell product prepared in the same manner was sealed in a desiccant-free packaging material bag. After encapsulating, the cells were allowed to stand at room temperature in the dark at 37 ° C., and the viable cell counts were measured after 1 week and 2 weeks, and the survival rate relative to the viable cell counts measured before encapsulation was calculated. As a control, a dry cell of a cell that was not enclosed in a bag (open air) was also measured, and the survival rate was calculated.

  When the desiccant-containing packaging material was used, the survival rate after one week was 87.1%, and after 7 weeks it was 77.4%. In the desiccant-free packaging material, they were 38.7% and 21.6%, respectively. When a desiccant-containing packaging material is used, the survival rate is good.

[Example 2]
Bacterial dried product 7 and a dried cell product obtained by removing 1% NaCl from the formula of dried cell product 7 (that is, a fungus obtained by lyophilizing 40% trehalose mixed with the bacterial cell) The dried body) was encapsulated in bags of desiccant-containing packaging materials ("Moist Catch (trade name)", manufactured by Kyodo Printing Co., Ltd.). After encapsulation, the sample was left in a room dark place at 37 ° C., and the viable cell count after 1 week was measured, and the survival rate relative to the viable cell count measured before encapsulation was calculated.

The survival rate after one week was 82.9% for the formulation containing 40% trehalose and 1% sodium chloride, and 66.4% for the formulation containing 40% trehalose alone.
The prescription with only 40% trehalose gave a good survival rate, and the addition of 1% sodium chloride further improved the survival rate.

[Example 3]
1 to 720 parts by weight of dried microbial cells, 75 parts by weight of calcium sulfate dihydrate and 5 parts by weight of naphthalenesulfonic acid (sodium) formaldehyde condensate were uniformly mixed to obtain a wettable powder composition. The obtained wettable powder composition was enclosed in a bag of a desiccant-containing packaging material (“Moist Catch (trade name)”, manufactured by Kyodo Printing Co., Ltd.). The viable cell count after 1 week was measured, and the survival rate relative to the viable cell count measured before encapsulation was calculated. Further, as a control, the survival rate was also calculated for the dried microbial cells that were not formulated.

  The obtained wettable powder composition had a cell survival rate of 83%, and the decrease in the number of cells due to the preparation was slight.

Claims (5)

A method for preserving a microbial pesticide preparation, comprising using a packaging material containing a desiccant. Microbial pesticide formulation
(A) It is a microbial pesticide preparation containing a dried microorganism cell product obtained by mixing sodium chloride and / or potassium chloride and / or (B) trehalose or sucrose and freeze-drying. The storage method according to claim 1. The preservation method according to claim 1 or 2, wherein the microorganism used in the microbial pesticide preparation is Pseudomonas rhodesia having an ability to control plant diseases. 4. The preservation method according to claim 3, wherein the Pseudomonas rhodesia having the ability to control plant diseases is Pseudomonas rhodesia 050572I9 strain (International Accession Number FERM BP-10912). The storage method according to claim 1, wherein the packaging material has a bag shape.