CN106305626A - Artificial culture medium for entomopathogenic nematodes - Google Patents
Artificial culture medium for entomopathogenic nematodes Download PDFInfo
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- CN106305626A CN106305626A CN201610692059.0A CN201610692059A CN106305626A CN 106305626 A CN106305626 A CN 106305626A CN 201610692059 A CN201610692059 A CN 201610692059A CN 106305626 A CN106305626 A CN 106305626A
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- Prior art keywords
- powder
- nutrient agar
- agar
- animal tissue
- entomopathogenic nematode
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000244206 Nematoda Species 0.000 title claims abstract description 23
- 230000000967 entomopathogenic effect Effects 0.000 title claims abstract description 18
- 239000001963 growth medium Substances 0.000 title abstract description 15
- 239000000843 powder Substances 0.000 claims abstract description 34
- 239000006916 nutrient agar Substances 0.000 claims abstract description 27
- 239000002609 medium Substances 0.000 claims abstract description 14
- 241001465754 Metazoa Species 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 10
- 210000002969 egg yolk Anatomy 0.000 claims abstract description 10
- 229920001817 Agar Polymers 0.000 claims abstract description 9
- 239000008272 agar Substances 0.000 claims abstract description 9
- 239000005645 nematicide Substances 0.000 claims description 24
- 241000282894 Sus scrofa domesticus Species 0.000 claims description 9
- 150000002333 glycines Chemical class 0.000 claims description 9
- 210000000582 semen Anatomy 0.000 claims description 9
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 238000004659 sterilization and disinfection Methods 0.000 claims description 9
- 241000254109 Tenebrio molitor Species 0.000 claims description 7
- 239000002184 metal Substances 0.000 claims description 5
- 230000002538 fungal effect Effects 0.000 claims description 4
- 239000010977 jade Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 12
- 239000007787 solid Substances 0.000 abstract description 10
- 230000008901 benefit Effects 0.000 abstract description 3
- 244000046052 Phaseolus vulgaris Species 0.000 abstract 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 abstract 1
- 238000012258 culturing Methods 0.000 abstract 1
- 235000013312 flour Nutrition 0.000 abstract 1
- 230000001524 infective effect Effects 0.000 abstract 1
- 239000010699 lard oil Substances 0.000 abstract 1
- 235000021048 nutrient requirements Nutrition 0.000 abstract 1
- 238000012136 culture method Methods 0.000 description 8
- 239000002917 insecticide Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 241000607479 Yersinia pestis Species 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 230000007918 pathogenicity Effects 0.000 description 3
- 241000239183 Filaria Species 0.000 description 2
- 201000006353 Filariasis Diseases 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 241001465746 Heterorhabditidae Species 0.000 description 1
- 241001523406 Heterorhabditis Species 0.000 description 1
- 241000500097 Heterorhabditis indica Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241001480238 Steinernema Species 0.000 description 1
- 241001480223 Steinernema carpocapsae Species 0.000 description 1
- 241001465745 Steinernematidae Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000012774 insulation material Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 235000006180 nutrition needs Nutrition 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- 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
- C12N1/20—Bacteria; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Animal Behavior & Ethology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses an artificial culture medium for entomopathogenic nematodes. The artificial culture medium is characterized by comprising a nutrient agar medium, wherein the nutrient agar medium is prepared from the following components: 25 to 50 g of animal tissue homogenate, 80 to 150 g of bean flour, 120 to 200 g of lard oil, 60 to 200 g of yolk powder, 8 to 15 g of agar, 12 to 20 g of yeast powder and 800 to 1200 ml of water. The artificial culture medium has the advantages that for a solid culturing method, nutrient requirements of different nematodes as well as convenience and cost of medium sources are considered, so that the yield of infective nematodes can be improved, and the production cost is reduced.
Description
[technical field]
The present invention relates to the technical field that biological self reproducing is cultivated, the particularly technology of the synthetic medium of entomopathogenic nematode
Field.
[background technology]
The genus steinernema Steinernema nematicide of Steinernema Carpocapsae section Steinernematidae and heterorhabditis indica
The mass production Heterorhabditis nematicide of Heterorhabditidae is entomopathogenic nematode, and they are current states
Biological insecticides novel on border.Compared with other biological insecticides, this kind of nematicide has host range widely；Host is had
Actively search capability, especially to dwelling property of soil and borer pest；To people and animals, plant and beneficial organism safety, in American-European some
Country can exempt registration and use；It is prone to mass propgation；Easy to use；Reusable edible in the environment；Can with multiple chemistry and
Biological insecticides are used with；And the advantage such as can use with tradition medical instruments or irrigating facility.
In recent years, as new bio insecticide, entomopathogenic nematode is widely used to agricultural, herbage and sanitary insect pest
Safe prevention, paid much attention to by Chinese scholars and commercial department, and moved towards merchandized handling.In industrialized country
In biological pesticide market, market sales revenue row's biological insecticides second of this type of nematicide.The whole world is more than 60 states at present
More than 100 laboratory of family is studying this type of beneficial organism.In China, entomopathogenic nematode has been commercially produced, and should
For preventing and treating multiple agricultural, meadow, flowers and sanitary insect pest etc., result is encouraging.
The cultivation of entomopathogenic nematode experienced by the process cultivating isolated culture from live body, and isolated culture is again from aseptic training
Support single bacterium to cultivate.At present, commercially producing of nematicide mainly adds list by aseptic technique in synthetic medium
One symbiotic bacteria and aseptic nematicide complete, the most external single bacterium culture systems.Solid culture and liquid can be divided into according to culture matrix
Body fermentation culture, both are not quite similar technically, such as culture vessel, Medium Proportion, incubation etc..
[summary of the invention]
The purpose of the present invention solves the problems of the prior art exactly, proposes the artificial culture of a kind of entomopathogenic nematode
Base, it is considered to the different nutritional needs of nematicide, the facility in culture medium source and cost, it is possible to increase infect phase nematode production, reduce
Production cost.
For achieving the above object, the present invention proposes the synthetic medium of a kind of entomopathogenic nematode, including Nutrient agar
Culture medium, the component of nutrient agar is: animal tissue homogenate 25g～50g, Semen Glycines powder 80～150g, Adeps Sus domestica 120～
200g, yolk powder 60～200g, agar 8～15g, yeast powder 12～20g, water 800ml～1200ml.
As preferably, the component of described nutrient agar is: animal tissue homogenate 40g, Semen Glycines powder 100g, Adeps Sus domestica
140g, yolk powder 100g, agar 10g, yeast powder 15g, water 900ml.
As preferably, the component of described nutrient agar is: animal tissue homogenate 35g, Semen Glycines powder 120g, Adeps Sus domestica
150g, yolk powder 150g, agar 12g, yeast powder 16g, water 1100ml.
As preferably, the nutrient agar that the using method of described nutrient agar is the most deployed is equal
Even it is mixed into the broken sponge of 120g, loads Vented metal and cultivate box, carry out high temperature sterilize sterilization, cool down stand-by；By cultured symbiosis
Bacterium is accessed in the nutrient agar after sterilization, inoculates nematicide after cultivating 4 days.
Beneficial effects of the present invention: the present invention is directed to solid culture method, the selection of culture medium prescription considers different nematicide
Nutritional need, culture medium source facility and cost.The production cost liquid culture side to be significantly lower than of solid culture method
Method, the Pathogenicity that solid culture method produces also will be apparently higher than liquid cultivating method.Therefore, increase when market demand
Greatly, solid culture method more industrialization production development prospect, the large-scale production to entomopathogenic nematode of the present invention has
Important meaning, the tenebrio molitor dry powder in culture medium can make the annealing time of nematicide one-level kind extend more than one times, it is possible to carries
Height infects phase nematode production, reduces production cost, good development prospect.
Inventive feature and advantage will be described in detail by embodiment.
[detailed description of the invention]
The synthetic medium of a kind of entomopathogenic nematode of the present invention, including nutrient agar, nutrient agar
Component be: animal tissue homogenate 25g～50g, Semen Glycines powder 80～150g, Adeps Sus domestica 120～200g, yolk powder 60～200g, agar 8
～15g, yeast powder 12～20g, water 800ml～1200ml.
Tenebrio molitor dry powder, the ultra-clean water of wet concentration are selected in the homogenate of described animal tissue.The use of described nutrient agar
The nutrient agar that method is the most deployed is uniformly blended into the broken sponge of 120g, loads Vented metal and cultivates box, carries out
High temperature sterilize is sterilized, and cools down stand-by；Cultured fungal component is accessed in the nutrient agar after sterilization, after cultivating 4 days
Inoculation nematicide.
Embodiment 1:
The component of nutrient agar is: animal tissue homogenate 40g, Semen Glycines powder 100g, Adeps Sus domestica 140g, yolk powder 100g,
Agar 10g, yeast powder 15g, water 900ml.
It is broken that the nutrient agar that the using method of nutrient agar is the most deployed is uniformly blended into 120g
Sponge, loads Vented metal and cultivates box, carry out high temperature sterilize sterilization, cool down stand-by；
Determining suitable cultivation temperature, scope is 19～28 DEG C, and cultured fungal component is accessed the nutrition fine jade after sterilization
In fat culture medium, after cultivating 4 days, inoculate nematicide.Often cultivate the optimum inoculation amount of 100,000,000 filarias at 200,000 tails～600,000 tails.
Embodiment 2:
As preferably, the component of described nutrient agar is: animal tissue homogenate 35g, Semen Glycines powder 120g, Adeps Sus domestica
150g, yolk powder 150g, agar 12g, yeast powder 16g, water 1100ml.
It is broken that the nutrient agar that the using method of nutrient agar is the most deployed is uniformly blended into 120g
Sponge, loads Vented metal and cultivates box, carry out high temperature sterilize sterilization, cool down stand-by；
Determining suitable cultivation temperature, scope is 19～28 DEG C, and cultured fungal component is accessed the nutrition fine jade after sterilization
In fat culture medium, after cultivating 4 days, inoculate nematicide.Often cultivate the optimum inoculation amount of 100,000,000 filarias at 200,000 tails～600,000 tails.
The technique effect that the present invention reaches
The quality standard of nematicide product
Regularity: the ratio that nematicide is infected under anatomical lens phase nematicide detects, uses the culture medium of the present invention
Phase of the infecting nematicide produced can reach more than 95%.
Pathogenicity: using tenebrio molitor as infecting object, infects 96 hours tenebrio molitor at 25 ± 1 DEG C, tenebrio molitor
Mortality rate can reach more than 80%.
Survival rate: finished product nematicide is packaged as kieselguhr powder absorption packaging, and with insulation material, the survival rate of nematicide is permissible
Reach more than 90%.
Nematicide one-level kind worm, after too much generation breeding, can run into the problem that kind of worm degenerates, be embodied in nematicide vigor, product
Amount declines, and the tenebrio molitor dry powder in culture medium can make the annealing time of nematicide one-level kind extend more than one times.
The present invention is directed to solid culture method, the selection of culture medium prescription considers the different nutritional need of nematicide, culture medium
The facility in source and cost.The production cost liquid cultivating method to be significantly lower than of solid culture method, solid culture method is raw
The Pathogenicity produced also will be apparently higher than liquid cultivating method.Therefore, when market demand increases, and solid culture method is more
Industrialization production development prospect, the large-scale production to entomopathogenic nematode of the present invention has great importance, it is possible to increase
Infect phase nematode production, reduce production cost, good development prospect.
Above-described embodiment is the description of the invention, is not limitation of the invention, any to simple transformation of the present invention after
Scheme belong to protection scope of the present invention.
Claims (5)
1. the synthetic medium of an entomopathogenic nematode, it is characterised in that: including nutrient agar, Nutrient agar is cultivated
The component of base is: animal tissue homogenate 25g～50g, Semen Glycines powder 80～150g, Adeps Sus domestica 120～200g, yolk powder 60～200g, fine jade
Fat 8～15g, yeast powder 12～20g, water 800ml～1200ml.
The synthetic medium of a kind of entomopathogenic nematode the most as claimed in claim 1, it is characterised in that: described animal tissue is homogenized
Selecting tenebrio molitor dry powder, wet concentration is with aseptic ultra-clean water.
The synthetic medium of a kind of entomopathogenic nematode the most as claimed in claim 1, it is characterised in that: described Nutrient agar is cultivated
The component of base is: animal tissue homogenate 40g, Semen Glycines powder 100g, Adeps Sus domestica 140g, yolk powder 100g, agar 10g, yeast powder 15g, water
900ml。
The synthetic medium of a kind of entomopathogenic nematode the most as claimed in claim 1, it is characterised in that: described Nutrient agar is cultivated
The component of base is: animal tissue homogenate 35g, Semen Glycines powder 120g, Adeps Sus domestica 150g, yolk powder 150g, agar 12g, yeast powder 16g, water
1100ml。
The synthetic medium of a kind of entomopathogenic nematode the most as claimed in claim 1, it is characterised in that: described Nutrient agar is cultivated
The nutrient agar that the using method of base is the most deployed is uniformly blended into the broken sponge of 120g, loads Vented metal and cultivates
Box, carries out high temperature sterilize sterilization, cools down stand-by；Cultured fungal component is accessed in the nutrient agar after sterilization, training
Nematicide is inoculated after supporting 4 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610692059.0A CN106305626A (en) | 2016-08-18 | 2016-08-18 | Artificial culture medium for entomopathogenic nematodes |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610692059.0A CN106305626A (en) | 2016-08-18 | 2016-08-18 | Artificial culture medium for entomopathogenic nematodes |
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| Publication Number | Publication Date |
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| CN106305626A true CN106305626A (en) | 2017-01-11 |
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| CN201610692059.0A Pending CN106305626A (en) | 2016-08-18 | 2016-08-18 | Artificial culture medium for entomopathogenic nematodes |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169534A (en) * | 2018-10-15 | 2019-01-11 | 南开大学 | Nematode solid medium and its preparation method and application |
| CN109430247A (en) * | 2018-11-21 | 2019-03-08 | 河南科技学院 | A kind of store method of Insectivorous nematode moisturizer and Insectivorous nematode |
| CN109496988A (en) * | 2018-11-01 | 2019-03-22 | 李小龙 | A kind of method and culture apparatus of entomopathogenic nematode pilot scale culture |
| CN110476896A (en) * | 2019-08-16 | 2019-11-22 | 潍坊宏润农业科技有限公司 | The cultural method and culture medium of Insect Pathogenic format nematode |
| CN115720963A (en) * | 2023-01-09 | 2023-03-03 | 北京林业大学 | Culture medium and culture method for Xindu nematode |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101411405A (en) * | 2008-11-26 | 2009-04-22 | 中国农业大学 | Insect tissue zymolyte and use thereof in artificial culture of entomopathogenic nematodes |
-
2016
- 2016-08-18 CN CN201610692059.0A patent/CN106305626A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101411405A (en) * | 2008-11-26 | 2009-04-22 | 中国农业大学 | Insect tissue zymolyte and use thereof in artificial culture of entomopathogenic nematodes |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169534A (en) * | 2018-10-15 | 2019-01-11 | 南开大学 | Nematode solid medium and its preparation method and application |
| CN109496988A (en) * | 2018-11-01 | 2019-03-22 | 李小龙 | A kind of method and culture apparatus of entomopathogenic nematode pilot scale culture |
| CN109496988B (en) * | 2018-11-01 | 2021-06-29 | 李小龙 | Method and device for large-scale culture of entomopathogenic nematodes |
| CN109430247A (en) * | 2018-11-21 | 2019-03-08 | 河南科技学院 | A kind of store method of Insectivorous nematode moisturizer and Insectivorous nematode |
| CN110476896A (en) * | 2019-08-16 | 2019-11-22 | 潍坊宏润农业科技有限公司 | The cultural method and culture medium of Insect Pathogenic format nematode |
| CN115720963A (en) * | 2023-01-09 | 2023-03-03 | 北京林业大学 | Culture medium and culture method for Xindu nematode |
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Application publication date: 20170111 |