CN1756482A - Process for producing granular nematocide - Google Patents

Process for producing granular nematocide Download PDF

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Publication number
CN1756482A
CN1756482A CNA2004800058098A CN200480005809A CN1756482A CN 1756482 A CN1756482 A CN 1756482A CN A2004800058098 A CNA2004800058098 A CN A2004800058098A CN 200480005809 A CN200480005809 A CN 200480005809A CN 1756482 A CN1756482 A CN 1756482A
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China
Prior art keywords
bacterium
granular
nematocide
nematode
medium
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CNA2004800058098A
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门宏超
山本惠理奈
菅沼修一
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JAPAN APPLIED MICROBIOLOGY RES
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JAPAN APPLIED MICROBIOLOGY RES
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A01N63/34—Aspergillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • Mycology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Botany (AREA)
  • Agronomy & Crop Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

It is intended to provide a process for producing a granular nematocide which makes it possible to preserve a microorganism having an nematocidal effect on pinewood nematode, root-knot nematode, cyst nematode, etc. over a long time. Namely, a process for producing a granular nematocide characterized by comprising culturing a microorganism having an nematocidal effect in a solid medium and then mixing the thus obtained culture with a granular carrier.

Description

The manufacture method of granular nematocide
Technical field
But the present invention relates to the bacterium of nematocidal effect of the manufacture method of granular nematocide long preservation has to(for) pine wood nematode, root-knot nematode, packing nematode, pratylenchus (キ ネ グ サ レ セ Application チ ユ ウ) etc.
Background technology
For example: put down in writing the bacterium that belongs to aspergillus (Aspergillussp.) in the special fair 2-32251 communique, this bacterium is as for being colonizing in pine wood nematode on the pine tree, colonize in root-knot nematode on cucumber or the cabbage, colonizing in the bacterium that packing nematode on the potato has nematocidal effect of vector insect with the ponderous borer.But, in above-mentioned patent documentation, its preparation is not carried out detailed research, investigate other documents and do not find corresponding report yet.
Therefore, but the purpose of this invention is to provide the bacterium of nematocidal effect of the manufacture method of granular nematocide long preservation has to(for) pine wood nematode, root-knot nematode, packing nematode, pratylenchus etc.
Summary of the invention
Point in view of the above problems, the manufacture method of nematocide of the present invention is characterized in that, have the bacterium of nematocidal effect as in solid culture medium, cultivating of putting down in writing of claim 1 after, bacterium culture and the bead-type substrate that obtains mixed.
In addition, the manufacture method of claim 2 record, it is characterized in that, in the manufacture method of claim 1 record, the bacterium with nematocidal effect is the bacterium that at least a nematode in pine wood nematode, root-knot nematode, packing nematode, the pratylenchus that belongs to Eurotium (Aspergillussp.) has nematocidal effect.
In addition, the manufacture method of claim 3 record is characterized in that in the manufacture method of claim 1 record, bead-type substrate is granular vermiculite.
In addition, granular nematocide of the present invention is characterized in that, this granular nematocide is made up of cultivate in solid culture medium resulting bacterium culture of bacterium and the granular vermiculite with nematocidal effect put down in writing as claim 4.
According to the present invention, but provide the manufacture method of the granular nematocide of the nematocidal effect of bacterium long preservation has to(for) pine wood nematode, root-knot nematode, packing nematode etc.
Description of drawings
Fig. 1 represents the life span evaluation test result of effective bacterium of 4 kinds of granular nematocides of embodiment 1.
Embodiment
The manufacture method of granular nematocide of the present invention is characterized in that, after cultivation has the bacterium of nematocidal effect in solid culture medium, bacterium culture and the bead-type substrate that obtains is mixed.According to the present invention, can make can the long preservation bacterium granular nematocide.
For can the long preservation bacterium, wish to keep bacterium to mix with spore state and carrier.In the present invention, the operation of cultivating the bacterium with nematocidal effect in solid culture medium can promote the sporulation of bacterium.Can be with the material known that is suitable for the Object of Development bacterium or condition as solid culture medium or condition of culture.As solid culture medium, can enumerate: the PDB (potatostarch that in the crushed materials such as carbide of vermiculite, nacrite, peat moss, plant corpus, adds the necessary nutrient component of spore of water, formation bacterium at least at least, dextrose, broth) with the material of potato block (rice bran also can) or add rice bran (wheat bran also can) with in requisition for various inorganic constituents (NaNO 3, K 2HPO 4, MgSO 4, KCl, FeSO 4) or the material of malt extract or peptone or glucose.The amount of the nutrient component in solid culture medium, use can promote the general quantity of the sporulation of bacterium to get final product.Particularly,, can enumerate as solid culture medium: contain the medium of PDB1 weight %～1.5 weight % and rice bran (wheat bran also can) 10 weight %～30 weight %, or contain in requisition for various inorganic constituents (NaNO 3, K 2HP 4, MgSO 4, KCl, FeSO 4) or the medium of malt extract or glucose or peptone.Under 20 ℃～30 ℃, to cultivate 10 days～28 days, the bacterium number of cultivating in solid culture medium reaches 10 8Cfu/g～10 9Get final product about cfu/g.
In addition, the bacterium of cultivating in solid culture medium with nematocidal effect also can be the bacterium culture of cultivating in known liquid nutrient medium in advance.But, if direct and bead-type substrate mixing with the bacterium culture of cultivating in the liquid medium within, because be difficult to form the spore of bacterium in the liquid nutrient medium, so can not obtain containing the preparation of a large amount of bacterium, having can not the long preservation bacterium or need be used for the problem of the drying process that the water that liquid nutrient medium is contained removes during preparation.So bacterium must mix with bead-type substrate after solid culture medium is cultivated again.When liquid medium within was cultivated in advance with bacterium, used liquid nutrient medium or condition of culture can adopt the known condition that is suitable for the Object of Development bacterium.For example as liquid nutrient medium, water with relative 100 weight portions, the PDB medium that contains the PDB of 2 weight portions～3 weight portions, the water of perhaps relative 100 weight portions contains the malt extract dextrose peptone medium of malt extract 1 weight portion～3 weight portions, glucose 1 weight portion～3 weight portions, peptone 0.05 weight portion～0.3 weight portion.Cultivation can be under 20 ℃～35 ℃, and concussion is cultivated and got final product in 3 days～10 days.
The bead-type substrate of the bacterium culture of being cultivated as the hybrid solid medium, can enumerate: the shot-like particle of the carbide of vermiculite, nacrite, peat moss, plant corpus etc., the effective bacterium that does not coexist from can long preservation assorted bacterium, promptly only the bacterium this point of long preservation with nematocidal effect is set out, and preferably uses granular vermiculite.In addition, the particle diameter of wishing bead-type substrate is 1 μ m～7mm.
For example, can mix bacterium and the bead-type substrate of cultivating with solid culture medium with mixed powder or the normally used mixing and blending machine of shot-like particle.Also the two or either party can be ground in advance and granularly mix again, also can be when mixing, pulverize make it to become granular.The mixed proportion of the two depends on the bacterium number that contains in the bacterium culture, wish to set for contain desired amt in the granular pesticide that makes manufacturing bacterium (for example: 5 * 10 5Cfu/cm 3～10 9Cfu/cm 3).
As the bacterium with nematocidal effect, preferred use belongs to the bacterium of aspergillus (Aspergillussp.), and this bacterium is the bacterium that has nematocidal effect for pine wood nematode, root-knot nematode, packing nematode, pratylenchus etc.Specifically can enumerate: honey aspergillus JAM4022 (Aspergillusmelleus JAM4022, FERM BP-726), in addition, also can use aspergillus parasiticus JAM4002 (Aspergillus parasiticus JAM4002, FERM BP-724), tower wood Richter scale aspergillus JAM4007 (Aspergillus tamarii JAM4007, FERM BP-723), aspergillus fumigatus JAM4008 (Aspergillus fumigatus JAM4008, FERM BP-727), aspergillus niger JAM4054 (Aspergillus niger JAM4054, FERM BP-8300), aspergillus niger JAM4055 (Aspergillus niger JAM4055, FERM BP-8301) etc.Wherein, JAM refer to Japan Applied Microbiology Res strain classification number ( JAPAN APPLIDE RESEARCH MThe abbreviation of ICROORGANISMS).
Embodiment
Below, describe the present invention in detail by embodiment, but, the present invention is not limited to this.
Embodiment 1
Production Example:
(operation 1)
To inoculate honey aspergillus JAM4022 (Aspergillus melleus JAM4022, FERMBP-726) spore has carried out PDA medium (PDA medium 39 weight portions of day water drugmaker manufacturing of preceding cultivation, distilled water 1000 weight portions) be inoculated in the PDB medium (PDB medium 2.4 weight portions, distilled water 100 weight portions that DIFCO company makes), cultivated 7 days 25 ℃ of concussions.The bacterium culture that cultivation in the liquid medium within of 1 weight portion is obtained is inoculated in the solid culture medium (pulverizing nacrite 63 weight portions of gardening company manufacturing, potato block 63 weight portions, distilled water 100 weight portions that ギ ャ バ Application ス バ ィ ス company makes are seen by PDB medium 2.4 weight portions, the Fuji that DIFCO company makes) of 9 weight portions, cultivates 14 days at 25 ℃.When mycelia and solid culture medium formation aggegation piece, agitation as appropriate makes it to disperse.Having confirmed that solid culture medium is cultivated in the granular bacterium culture that obtains contains 10 8The above effective bacterium of cfu/g (according to the Agar Plating dilution method, below identical).
(operation 2)
At normal temperatures, about the granular bacterium culture and the about 50 μ m of particle diameter that obtains with mixing and blending machine mixed processes 1～and about 4 kinds of bead-type substrates (the granular carbide of granular vermiculite, granular nacrite, granular peat moss, plant corpus) about 5mm, make granular nematocide.Set the mixed proportion of the two so that contain 5 * 10 in the granular nematocide of making 5Cfu/cm 3Above effective bacterium.
The test example:
Different granular nematocides with 4 kinds of bead-type substrate manufacturings of embodiment 1 are carried out the life span test evaluation of effective bacterium of 2 months by a definite date under 25 ℃.The result as shown in Figure 1.Can find out clearly that from Fig. 1 the granular carbide with granular nacrite and plant corpus reduces as the bacterium digital display of the granular nematocide of bead-type substrate is outstanding, with vermiculite and granular peat moss granular nematocide as bead-type substrate, since on-test, even passed through the minimizing of also almost not finding existence bacterium number in 8 months.But, with containing a lot of assorted bacterium in the granular nematocide of peat moss as bead-type substrate.
Embodiment 2
Production Example:
(operation 1)
With inoculated aspergillus niger JAM4055 (Aspergillus niger JAM4055, FERMBP-8301) spore has carried out PDA medium (PDA medium 39 weight portions of day water drugmaker manufacturing of preceding cultivation, distilled water 1000 weight portions) be inoculated in the liquid nutrient medium (peptone 1 weight portion, glucose 20 weight portions, distilled water 1000 weight portions of malt extract 20 weight portions that オ リ エ Application Le yeast industry company makes, utmost point Dong Zhiyaoshe preparation), cultivated 7 days 25 ℃ of concussions.Solid culture medium (the NaNO that the bacterium culture that obtains is inoculated into 16 weight portions will be cultivated in the liquid medium within of 1 weight portion 32.25 weight portion, K 2HPO 40.75 weight portion, MgSO 47H 2O 0.325 weight portion, KCl 0.325 weight portion, FeSO 47H 2Pulverizing nacrite 500 weight portions, rice bran 500 weight portions, moisture adjusting rice husk 165 weight portions, distilled water 750 weight portions that gardening company makes are seen by O0.0075 weight portion, sucrose 22.5 weight portions, Fuji) in, cultivated 14 days at 25 ℃.When mycelia and solid culture medium formation aggegation piece, agitation as appropriate makes it to disperse.Having confirmed that solid culture medium is cultivated in the granular bacterium culture that obtains contains 10 8Effective bacterium that cfu/g is above.
(operation 2)
At normal temperatures, about the granular bacterium culture and the about 50 μ m of particle diameter that obtains with mixing and blending machine mixed processes 1～the about granular vermiculite about 5mm, make granular nematocide.The mixed proportion of the two is set so that contain 5 * 10 in the granular nematocide of making 5Cfu/cm 3Above effective bacterium.
The test example:
The granular nematocide that embodiment 2 is made carries out the life span test of effective bacterium of 2 months by a definite date under 25 ℃, it is estimated, found that, even passed through time of 6 months from on-test, almost do not find the minimizing of existence bacterium number yet, and contain assorted bacterium hardly.
Utilizability on the industry
But the invention provides long preservation for pine wood nematode, root-knot nematode, packing nematode etc. The manufacture method of granular nematocide with bacterium of nematocidal effect. Consider from above-mentioned starting point, The present invention has utilizability industrially.

Claims (4)

1. the manufacture method of granular nematocide is characterized in that, after cultivation has the bacterium of nematocidal effect in solid culture medium, bacterium culture and the bead-type substrate that obtains is mixed.
2. the manufacture method of claim 1 record, it is characterized in that, this Pseudomonas with nematocidal effect is in Eurotium (Aspergillus sp.), and this bacterium is for being selected from the bacterium that at least a nematode in pine wood nematode, root-knot nematode, the packing nematode has nematocidal effect.
3. the manufacture method of claim 1 record is characterized in that bead-type substrate is granular vermiculite.
4. granular nematocide is characterized in that, is formed by cultivate resulting bacterium culture of bacterium and granular vermiculite with nematocidal effect in solid culture medium.
CNA2004800058098A 2003-03-04 2004-03-03 Process for producing granular nematocide Pending CN1756482A (en)

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JP2003057481A JP2006182647A (en) 2003-03-04 2003-03-04 Method for producing granular nematocide
JP57481/2003 2003-03-04

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016163726A1 (en) * 2015-04-06 2016-10-13 전남대학교산학협력단 Aspergillus niger f22 strain having nematicidal activity against plant-parasitic nematodes, and use thereof

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DE102005031336B4 (en) 2005-05-13 2008-01-31 Osram Opto Semiconductors Gmbh projection device

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JPS61231933A (en) * 1985-04-04 1986-10-16 堀内　勲 Control of worm of plant
JP3509056B2 (en) * 1998-04-20 2004-03-22 中川 彰 Microbial material that suppresses the growth of harmful soil nematodes and its production method

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016163726A1 (en) * 2015-04-06 2016-10-13 전남대학교산학협력단 Aspergillus niger f22 strain having nematicidal activity against plant-parasitic nematodes, and use thereof

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WO2004077946A1 (en) 2004-09-16
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