CN1928065A - Industrial high power breeding method for entomopathogenic nematode alive body - Google Patents
Industrial high power breeding method for entomopathogenic nematode alive body Download PDFInfo
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- CN1928065A CN1928065A CN 200610016563 CN200610016563A CN1928065A CN 1928065 A CN1928065 A CN 1928065A CN 200610016563 CN200610016563 CN 200610016563 CN 200610016563 A CN200610016563 A CN 200610016563A CN 1928065 A CN1928065 A CN 1928065A
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Abstract
The present invention belongs to the field of agricultural production technology, and is especially industrial high power living body breeding method of insect pathogen nematode. The present invention adopts living body breeding, rather than in vitro breeding, and has obviously raised agricultural pest invading power, high yield and efficiency, low cost, high quality and obviously raised pest preventing and controlling effect. The present invention may be used widely in preventing and controlling pest of potted crop, economic crop, vegetable, pasture, etc.
Description
Technical field
The invention belongs to field of agricultural production technologies.It is a kind of industrial high power breeding method for entomopathogenic nematode alive body.
Technical background
At present, chemical agent is mainly adopted in controlling underground pest and disguised pest control, but this method easily causes the person poultry poisoning, also pollutes the environment.And repeatedly prevent and treat the cost height, insect also is easy to generate resistance.Preparation is controlled in the raising that food safety is required along with people, and to the consideration that chemical agent may bring potential collision hazard to ecotope, the biological and ecological methods to prevent plant disease, pests, and erosion of screening alternative chemical agent to be become and presses for.
Filter out Bacillus thuringiensis (Bacillus thuringiensis), muscardine (Beauveriabrongniatii), green muscardine fungus (Metarhizium anisopliae) etc. now as the biological control preparation, but prevented and treated some subterranean pest-insects and disguised insect effect is not very desirable.And entomopathogenic nematode (Entomopathogenic Nematode) is the specialization parasite of insect.Enter in the insect body with host's food or from anus, pore, the intersegmental membrane of insect to infect phase worm attitude (promptly three age nematode IJ), discharge the symbiotic bacterium that it carries subsequently, these symbiotic bacteriums breed rapidly, and it is dead within 48 hours to make host insect suffer from septicemia.Entomopathogenic nematode is a kind of very ergastic biological control factor.Many countries are arranged in the world today, all drop into this nematode of great amount of manpower and material resources research and utilization, be devoted to the nematode production marketing to all over the world as countries such as the U.S., Australia, New Zealand, Germany, Egypt.1994 according to Lisansky ﹠amp; The Coombs statistics, in the biological pesticide market of developed country, the market sales revenue of this novel pesticide of entomopathogenic nematode is only second to industrial Tribactur, successfully has been applied to prevent and treat subterranean pest-insect and fruit trees such as potted plant crop, cash crop, vegetables and herbage, lawn, has viewed and admired disguised insect on the forests such as gardens.So this new bio sterilant of entomopathogenic nematode has broad application prospects, but used nematode population is few, can not meet the need of market.
Because entomopathogenic nematode can be cultivated breeding in a large number, it can be applied.Though the nematode infection power that the breeding of traditional White trap method obtains is higher, but its scale does not satisfy the demand in current market far away, Guangdong Korean and Japanese of insect institute adopts on the farmland method of artificial in-vitro propagate, the fermentor tank of nematode produced in USA is raised to Biosys company from 5 liters, 20 and produces 15000 liters of Stahli line worms, produce the scale of 7500 liters of heterorhabditis indicas, though nematode production is higher, its virulence descends to some extent.
Summary of the invention
The purpose of this invention is to provide a kind of industrial high power breeding method for entomopathogenic nematode alive body, is a large amount of high power living body propagation of batch production entomopathogenic nematode, reaches the good result of biological control insect pest in the agricultural production practice.Gordian technique of the present invention is to existing in-vitro propagate, changes living body propagation into, and nematode is obviously strengthened the virulence of Agricultural pests, and output and efficient are higher, and cost is low, and quality is good, and prevention effect obviously promotes.
The present invention mixes in the breeding bag of packing into host insect after adopting the developing medium inoculation to infect the phase nematode, will breed bag and place constant incubator to cultivate, and culture parameters is optimized, and will obtain a large amount of entomopathogenic nematodes.Concrete steps following (referring to accompanying drawing):
A. breeding bag quality screening: host insect is adopted tenebrio molitor, developing medium vermiculite.The vermiculite that buys from market soaks with tap water, and wash clean is dried to constant weight in 60 ℃ of baking ovens then.
Infect early-stage preparations: the 1Kg vermiculite is contained in is immersed in the cloth bag in the Distilled water bucket, after soaking fully, take out cloth bag, hang up, dry on cloth bag and do not drip, claim the weight of cloth bag again, calculate the vermiculite water content, promptly vermiculite soaks into institute's water requirement fully.
Infect:, be made into certain density nematode suspension the nematode suspension.Certain density nematode suspension is sprayed on the vermiculite, makes vermiculite drenched fully, do not have water to overflow and be advisable.
Cloth bag is 17 * 26cm size, and the cloth quality has nylon, cotton textiles, establish infect every last age tenebrio molitor heterorhabditis indica dosage be 50 IJ, 100 IJ of Stahli line worm.It is 5 times of Nylon Bag that the result breeds nematode production that bag adopts textile cloth to make.
B. developing medium screening: select local the acquisition easily and low-cost resource---vermiculite and perlite for use, host insect, developing medium handle and infective dose goes on foot with a, and the result is that the nematode production of developing medium is perlitic 3 times with the vermiculite.
C. whether developing medium sterilizes: host insect, developing medium handle and infective dose goes on foot with a, and the unsterilised nematode production of developing medium is higher than 1.25 times of sterilization as a result, needn't sterilize so consider the reproductive efficiency developing medium.
D. whether breeding bag stirs: host insect, developing medium handle and infective dose goes on foot with a, the result breed bag stir suitable with the nematode production that does not stir, so the breeding bag also needn't stir.
E. infective dose screening: host insect and developing medium blending ratio, harvest time, breeding bag size are carried out the Orthogonal Composite test, host insect and developing medium are handled the step with a, and host insect (tenebrio molitor) is three to two with developing medium (vermiculite) blending ratio as a result; The heterorhabditis indica infective dose is 800 and infects phase nematode/gram tenebrio molitor (Stahli line worm infective dose is 1600 and infects phase nematode/gram tenebrio molitor); Breeding bag size is selected 17 * 26cm for use; The harvest time of heterorhabditis indica is to begin results when cultivating 12 days, cloth bag is soaked and gather in the crops 5 times, gather in the crops once more when cultivating 15 days, cloth bag soaked and gather in the crops 3 times (harvest time of Stahli line worm is to begin results when cultivating 10 days, cloth bag is soaked and gather in the crops 5 times, gather in the crops once more when cultivating 13 days, cloth bag is soaked and gather in the crops 3 times);
F. harvesting method optimization: with nematode suspension precipitation, remove by filter impurity with the medicated napkin that is placed on the porcelain dish again after removing supernatant liquor after the results, can keep the original virulence of nematode, and the life-span is than 6 times of the nematode prolongations of not filtering storage.
G. batch production production: the best culture parameters that filters out more than the selection, breeding bag is suspended in 25 ℃ of rooms that air-conditioning is housed, observes the reading that being suspended in the steelyard on the breeding bag every day, with the fluid loss of monitoring breeding bag, when dehydration reaches 50%, use distilled water and squirt cloth bag.Finally obtain breeding rate height, the measured nematode of matter.
Used plant and instrument: DHP-9162A constant temperature culture phase (Tianjin Tai Site), 30405359 phase microscopes (Mike Audi), S633069 cold light source anatomical lens (Mike Audi), Z18400 micropipet (Shanghai).
Description of drawings: Fig. 1 is an entomopathogenic nematode living body propagation schema
Embodiment
The present invention is field, a Chinese Academy of Sciences subject forward position project: the main contents in " utilizing pathogenic nematode control soybean subterranean pest-insect and soy bean cyst roundworm rDNA-ITS-RFLP research " (numbering 20020101).We have carried out batch production experimental production in Northeast Institute of Geography and Agricultural Ecology, Chinese Academy of Sc by above step year December in January, 2002 to 2005, obtained the success of the best high power propagation method of entomopathogenic nematode live body, can solve the difficult problem that nematode population can not be applied to production practice less, reach more than 80% through overtesting control subterranean pest-insect effect, both obtain economic benefit, obtained ecological benefits again.
Materials used of the present invention distributes wide, and low price can reduce cost.Be widely used in controlling underground pest and fruit trees such as potted plant crop, cash crop, vegetables, herbage, lawn, view and admire disguised pest control on the forests such as gardens, the present invention will further improve the output and the quality of entomopathogenic nematode, promote the development of China's biological control.
Claims (1)
1. high power breeding method for entomopathogenic nematode alive body is produced in a batch production, it is characterized in that this present in-vitro propagate is a living body propagation, and its step is as follows:
A. breeding bag quality screening: host insect is adopted tenebrio molitor, developing medium vermiculite.The vermiculite that buys from market soaks with tap water, and wash clean is dried to constant weight in 60 ℃ of baking ovens then;
Infect early-stage preparations: the 1Kg vermiculite is contained in is immersed in the cloth bag in the Distilled water bucket, after soaking fully, take out cloth bag, hang up, dry on cloth bag and do not drip, claim the weight of cloth bag again, calculate the vermiculite water content, promptly vermiculite soaks into institute's water requirement fully;
Infect: with the nematode suspension, be made into certain density nematode suspension, certain density nematode suspension is sprayed on the vermiculite, make vermiculite drenched fully, do not have water to overflow and be advisable;
Cloth bag is 17 * 26cm size, and the cloth quality has nylon, cotton textiles, establish infect every last age tenebrio molitor heterorhabditis indica dosage be 50 IJ, 100 IJ of Stahli line worm;
B. developing medium screening: select local the acquisition easily and low-cost resource---vermiculite and perlite for use, host insect, developing medium handle and infective dose goes on foot with a;
C. whether developing medium sterilizes: host insect, developing medium handle and infective dose goes on foot with a, consider reproductive efficiency, and developing medium needn't be sterilized;
D. whether the breeding bag stirs: host insect, developing medium handle and infective dose goes on foot with a, and for improving reproductive efficiency, the breeding bag needn't stir;
E. infective dose screening: host insect and developing medium blending ratio, harvest time, breeding bag size are carried out the Orthogonal Composite test, host insect and developing medium are handled the step with a, and it is three to two with developing medium (vermiculite) blending ratio that effect is preferably host insect (tenebrio molitor); The heterorhabditis indica infective dose is 800 and infects phase nematode/gram tenebrio molitor (Stahli line worm infective dose is 1600 and infects phase nematode/gram tenebrio molitor); Breeding bag size is selected 17 * 26cm for use; The harvest time of heterorhabditis indica is to begin results when cultivating 12 days, cloth bag is soaked and gather in the crops 5 times, gather in the crops once more when cultivating 15 days, cloth bag soaked and gather in the crops 3 times (harvest time of Stahli line worm is to begin results when cultivating 10 days, cloth bag is soaked and gather in the crops 5 times, gather in the crops once more when cultivating 13 days, cloth bag is soaked and gather in the crops 3 times);
F. harvesting method optimization: with nematode suspension precipitation, remove by filter impurity with the medicated napkin that is placed on the porcelain dish again after removing supernatant liquor after the results, can keep the original virulence of nematode;
G. batch production production: the best culture parameters that filters out more than the selection, breeding bag is suspended in 25 ℃ of rooms that air-conditioning is housed, observes the reading that being suspended in the steelyard on the breeding bag every day, with the fluid loss of monitoring breeding bag, when dehydration reaches 50%, use distilled water and squirt cloth bag.
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| CN 200610016563 CN1928065A (en) | 2006-01-24 | 2006-01-24 | Industrial high power breeding method for entomopathogenic nematode alive body |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102640730A (en) * | 2012-04-13 | 2012-08-22 | 中国科学院东北地理与农业生态研究所 | In vivo reproduction method of entomopathogenic nematodes |
| CN103004726A (en) * | 2012-12-28 | 2013-04-03 | 甘肃农业大学 | Simple and effective method for trapping entomopathogenic nematodes |
| CN103053619A (en) * | 2011-10-20 | 2013-04-24 | 南开大学 | Antibacterial, anti-dry and anti-adhesion method of dead insects infected by entomopathogenetic nematodes |
| CN103053620A (en) * | 2011-10-20 | 2013-04-24 | 南开大学 | Entomopathogenic nematodes insect corpse agent |
| CN105532583A (en) * | 2015-12-29 | 2016-05-04 | 宁波市农业科学研究院 | Living body breeding method for entomopathogenic nematodes |
| CN108719208A (en) * | 2018-05-15 | 2018-11-02 | 吉林省怡科农业生物科技有限公司 | Entomopathogenic nematode large-scale method for producing and entomopathogenic nematode |
-
2006
- 2006-01-24 CN CN 200610016563 patent/CN1928065A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103053619A (en) * | 2011-10-20 | 2013-04-24 | 南开大学 | Antibacterial, anti-dry and anti-adhesion method of dead insects infected by entomopathogenetic nematodes |
| CN103053620A (en) * | 2011-10-20 | 2013-04-24 | 南开大学 | Entomopathogenic nematodes insect corpse agent |
| CN102640730A (en) * | 2012-04-13 | 2012-08-22 | 中国科学院东北地理与农业生态研究所 | In vivo reproduction method of entomopathogenic nematodes |
| CN103004726A (en) * | 2012-12-28 | 2013-04-03 | 甘肃农业大学 | Simple and effective method for trapping entomopathogenic nematodes |
| CN103004726B (en) * | 2012-12-28 | 2015-11-18 | 甘肃农业大学 | A kind of method for enticing collection of simple and effective entomopathogenic nematode |
| CN105532583A (en) * | 2015-12-29 | 2016-05-04 | 宁波市农业科学研究院 | Living body breeding method for entomopathogenic nematodes |
| CN105532583B (en) * | 2015-12-29 | 2018-04-06 | 宁波市农业科学研究院 | A kind of living body propagation method of entomopathogenic nematode |
| CN108719208A (en) * | 2018-05-15 | 2018-11-02 | 吉林省怡科农业生物科技有限公司 | Entomopathogenic nematode large-scale method for producing and entomopathogenic nematode |
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