CN1775942A - Predacious roundworm fungus bacterial strain and its preparing method - Google Patents
Predacious roundworm fungus bacterial strain and its preparing method Download PDFInfo
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- CN1775942A CN1775942A CN 200410090815 CN200410090815A CN1775942A CN 1775942 A CN1775942 A CN 1775942A CN 200410090815 CN200410090815 CN 200410090815 CN 200410090815 A CN200410090815 A CN 200410090815A CN 1775942 A CN1775942 A CN 1775942A
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Abstract
The invention relates to a method to make epiphyte strain that preys upon wireworm. It could be used to prevent plant wireworm and control infestation wireworm in intestinal tract of ruminant. It could decompose the fibrin and chitin of plant relict body in earth. If adding it into feedstuff, it would restrain the mildew. The microorganism of the invention is from earth, so it has no harm to plant, human and livestock.
Description
Technical field:
The present invention relates to microbial technology field, specifically a kind of predacious roundworm fungus bacterial strain and preparation method.
Technical background:
Plant nematode is one of important pathogen of farm crop, can cause crop failure 10～30%, have in addition the total crop failure.Wherein root knot nematode Meloidogyne, golden nematode Heterodera and Ditylenchus dipsaci Ditylenchus are 3 main nematode monoids that restriction is produced.To the control of plant nematode always based on crop rotation, cultivation step, use disease-resistant variety, use chemical agent.But cyst of nematode or the ovum survival time in soil is long, brings certain difficulty for crop rotation, cultivation step control.Disease-resistant variety often causes in an area the single resistant variety of plantation continuously, because the effect of kind selective pressure changes the colony of nematode, finally causes the varietal resistance forfeiture.In present actual production, the main means of controlling plant parasitic nematode harm are to use chemical nematode killing agent, and as aldicarb, furans pellet, various soil fumigants etc., they mostly are highly toxic pesticide greatly, the resistates of these products can remain in underground water and the soil, and environment is caused severe contamination; Because many crop products are as the direct food consumed of people, its residual medicine will directly threaten human beings'health on the other hand; In addition, because these products are poisonous to the people,, all forbid as a lot of chemical nematocidess such as the U.S., Holland, Germany in many developed countries.Utilizing the natural enemy of nematode to prevent and treat nematode is a kind of safe and effective biological control method, by the plant protection work person of countries in the world is favored.The natural enemy of nematode mainly contains fungi, bacterium, virus, predatism nematode, mite class, turbellarian worm, insect and protozoon etc., and wherein fungi has just accounted for 75%, is the important nematode controlling elements of occurring in nature.Though there is part nematode biocontrol fungi preparation to occur at present, but the most more complicated of preparation technology, simultaneously since in the edatope as the influence of soil fungistasis etc., it is very difficult to make the survival of biocontrol microorganisms in soil grow surely, especially can not produce the biocontrol microorganisms of degeneration-resistant thalline structure.
The animal parasitic nematode is one of important parasite that influences animal husbandry development.Wherein haemonchus (Haemonchus), esophageal orifice nematode (Oesophagostomum), Bunostomum trigonoce phalum (Bunostomum), oersted nematode (Ostertagia), Xia Baite nematode (Chabertia) and sheep net filaria (Dictyocaulus) are main parasitic nematodes.Its harm mainly is to cause the grow up decline of Livestock Production rate, malnutrition and circulatory disorder, and childhood domestic animal hypoevolutism etc.How to prevent and treat animal nematodiasiss and become one of key factor that improves livestock industry production.The method that is used at present preventing and treating the animal nematode both at home and abroad also mainly is based on promptly controlling nematode by chemicals in the use of chemical flooding vermicide.And the pasture is the important worm source of parasitic nematode, and the feed of nematode follower is taken in the body, and general medicament does not have long-term insecticidal action; But life-time service can make the nematode kind develop immunity to drugs; In addition, medicine residual can causing people's health directly or potential harm in animal body.Thereby people begin to seek from others the approach of control nematode, and biological control is exactly one of them.Wherein utilizing nematode-trapping fungi control animal parasitic nematode is a kind of main biological control method.Though the nematode-trapping fungi that has has higher predation rate to nematode, but owing to can not resist the digestive environments of animal digestive tract, thereby can not come the parasitic nematode in the environment is controlled by the method that feed adds, animal is fed easily, its use is restricted.
1998, Tepljakova T V; Zelenkov V N to plant, the application of animal parasitic nematode predacious fungi production technique patent (patent No. RU2106092).Preparation extracts mycelia then in a large number at first by nodal plexus spore (Arthrobotrys oligospora) BKMF-3062D liquid culture, and adds kaolin or zeolite as carrier by 1: 2 ratio, sieves at last, drying, makes the granule use.This patented strain and the granule of making are not described and can be used for adding as simple feed.
In addition, the same with other microorganism live body preparation, there is the short problem of preparation preservation period equally in the nematicide microbial preparation, generally at room temperature deposits more than 6 months, and its viable count often reduces more than 80%.
Therefore, the purpose of this invention is to provide a kind of can be used to simultaneously prevent and treat plant nematode and nematode-trapping fungi of controlling the intestine of ruminants parasitic nematode and preparation method thereof.
Therefore, one of purpose of the present invention just is to use a kind of nematode-trapping fungi control plant nematode from nature, and it is all harmless to people and environment.
The present invention relates to a kind of nematode-trapping fungi of preventing and treating Meloidogyne Meloidogyne, Heterodera Heterodera and Ditylenchus Ditylenchus destructor nematode and preparation method thereof.
Therefore, another object of the present invention just is to use a kind of nematode-trapping fungi from nature, the method of adding by feed makes the ruminating animal picked-up and is excreted to by enteron aisle controls the infection of animal parasitic nematode to ruminating animal in the environment, it is to animal, environment and harmless per capita.Nematode as controlled target mainly comprises haemonchus (Haemonchus), esophageal orifice nematode (Oesophagostomum), Bunostomum trigonoce phalum (Bunostomum), oersted nematode (Ostertagid), Xia Baite nematode (Chabertia) and sheep net filaria (Dictyocaulus).
Therefore, another object of the present invention just provide a kind of can be after preserving 1 year, make in the preparation effective microbe body burden above nematode-trapping fungi more than 50% and preparation method thereof.
Summary of the invention:
The biological control of harmful nematode utilizes antagonistic microbe to prevent exactly or reduces harmful nematode causing harm to plant and animal.The objective of the invention is at also there not being at present the very economical microbial preparation that effectively is used to prevent and treat harmful nematode, and provide a kind of specific nematode-trapping fungi that separation screening is turned out from soil, be used to prevent and treat plant nematode and intestine of ruminants parasitic nematode, the present invention also provides the preparation method of this nematode-trapping fungi.
For being separated to the nematode-trapping fungi strain effectively, the present invention is by making the method for bait and picking nematode-trapping fungi trapping organs with nematode, separate and obtain nematode-trapping fungi BJNF-1 every referring to spore (Dactylella sp.), this bacterial classification is in specified microbial preservation unit of Patent Office of the People's Republic of China, China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, its preservation registration number is: CGMCC No.1051.
BJNF-1 is every referring to that spore (Dactylella sp.) has following character:
Cultural characteristic:
On potato dextrose agar (PDA), 25 ℃, a large amount of three-dimensional predation bacterium nets be can form in 3 days, and conidium and chlamydospore produced, the aerial hyphae prosperity, cotton-shaped, bacterium colony is light brown, and 6 days colony diameter 75mm produced a large amount of chlamydospores in 10 days.
On Corn Meal Agar substratum (CMA), 25 ℃, bacteria colony white, mycelia crawls, and is colourless, and the bacterium colony mycelia is very fine and close, and the bacterium colony circle that keeps to the side produces conidium and a large amount of chlamydospores, and 6 days colony diameter 70mm produced a large amount of chlamydospores in 10 days.
On the yeast culture base, 25 ℃, 6 days colony diameter 70mm, bacterium colony front pale asphyxia, back side tawny, aerial hyphae is extremely flourishing, the carpet shape, the outer rim mycelia is comparatively sparse, is the straw hat shape, can produce a large amount of conidiums.
On nutrient agar (NA), 25 ℃, 6 days colony diameter 50mm, the positive mycelia of bacterium colony is undeveloped, sparse, white, the back side is light brown, produces the solubility brown pigments, and a small amount of generation of conidium is arranged.
The microscopy form:
Newborn mycelia does not have every, old mycelia and has every, wide 2.5～7.0 μ m of vegetative hyphae.
Conidiophore: Dan Sheng, upright, separate, few branch, long 250～300 μ m of stalk, wide 6.5～7.5 μ m of base portion, wide 4.0～5.5 μ m in top, the stalk top gives birth to 1～several conidiums, and the spore producing method is full wall budding formula, and does not expand on the sporophore top.
Conidium: intend bar-shapedly, the nearly base portion sudden turn of events is narrow, and like bottleneck, base portion is truncate, does not have and separates or 1 separation; Have and separate spore and separate generally on the lower side, the part spore bundle of hanging in the separated place at the middle part; The part nothing caves at the middle part slightly every conidium; The blunt circle in most conidium top, the blunt point of only a few, conidium size 35.0～45.0 * 9.0～11.0 μ m.
Chlamydospore: on artificial medium, form a large amount of chlamydospores, give birth between chlamydospore or push up and give birth to, can on substrate mycelium, form, also can produce on the net at three-dimensional slime bacteria, chlamydospore sphere or sphaeroid, tawny, ripe chlamydospore tool surface boss, size is 32.0～35.0 * 27.0～32.0 μ m
Trapping organs: be not really flourishing three-dimensional slime bacteria net, can when existing, no nematode on different artificial mediums, produce in a large number, three-dimensional bacterium netting gear has very strong nematode-trapping ability, can catch nematode when the bacterium net just forms mycelioid, can produce chlamydospore on the net bacterium.
Above-mentioned nematode-trapping fungi is separation and Culture by the following method:
Making bait with nematode is added on the nutrient agar that is connected to pedotheque, 25～28 ℃ of dark cultivations after 5～8 days, the predation bacterium net that produces is chosen to potato dextrose agar, substratum includes penicillin, Streptomycin sulphate and duomycin and is respectively 20 μ g/ml, 40 μ g/ml, 30 μ g/ml, cultivate this nematode-trapping fungi.
The preparation method of above-mentioned nematode-trapping fungi is characterized in that this preparation method carries out according to the following steps:
(1). test tube is cultivated: adopt potato dextrose agar, this bacterium is seeded on the test tube substratum, cultivated 3～5 days for 25～30 ℃.
(2). enlarged culturing: adopt liquid potato glucose substratum, be seeded in the Erlenmeyer flask of filling liquid substratum cultivating bacterium in the above-mentioned test tube, 25～30 ℃ of vibrations are secretly cultivated 5～6 days as the solid culture seed on shaking table.
(3). solid culture: adopt wheat straw bar potato glucose solid medium: dried wheat straw stalk is cut into the long stalk section of 2mm, and the wheat stalk section being absorbed water and make drains the water after saturated; Potato is boiled and makes its one-tenth pasty state with suitable quantity of water; Make dried wheat straw stalk, potato, glucose, three's mass ratio is 25: 10: 1, mixes, mixes evenly; 121 ℃ of high pressure steam sterilizations 30～60 minutes are cooled to room temperature.In mass ratio: solid medium: liquid seeds is 50: 1 a ratio, the enlarged culturing seed is inoculated on the solid medium, and mixing, 28 ℃ of dark cultivations obtained the nematode-trapping fungi solid culture in 5～7 days.
The above-mentioned gained nematode-trapping fungi solid culture chlamydospore content after through normal temperature (25 ℃) seasoning or 35 ℃ of artificial freeze-day with constant temperature reaches 10
6Individual every gram dry weight; After the nematode-trapping fungi solid culture was preserved 3 years through normal temperature (25 ℃) seasoning, its chlamydospore survival rate was 15%.The nematode-trapping fungi solid culture is made the nematode-trapping fungi preparation through certain technical process, is used to prevent and treat plant nematode and prevents or reduce the infection of intestine of ruminants parasitic nematode to ruminating animal.
Adopt nematode-trapping fungi of the present invention that plant nematode Meloidogyne Meloidogyne, Heterodera Heterodera in the soil and Ditylenchus Ditylenchus destructor nematode are prevented and treated, can utilize this bacterium to decompose the Mierocrystalline cellulose of plant residue in the soil, chitin etc. on the one hand, increase the nutrient composition content in the soil, promote soil with organic matter to transform; Can improve soil fertility again, the environmental pollution that minimizing chemical fertilizer, agricultural chemicals cause; This bacterium can also reduce the control cost thereby can carry out Sustainable Control to the nematode in the soil in the medium-term and long-term survival of soil.
Adopt nematode-trapping fungi of the present invention to the intestine of ruminants parasitic nematode, mainly comprise haemonchus (Haemonchus), esophageal orifice nematode (Oesophagostomum), Bunostomum trigonoce phalum (Bunostomum), oersted nematode (Ostertagia), summer Bert nematode (Chabertia) and sheep net filaria (Dictyocaulus) are prevented and treated, by adding the nematode-trapping fungi preparation in feed, the digestive tube by animal is excreted in the environment nematode is worked; Using method is easy on the one hand; By of the inhibition of this bacterium, can make the feed storage safer in addition, reduce the pollution of mould feed to mould.
Microorganism of the present invention is from anthropogenic soil, and to plant and person poultry harmless, easily production, anti-storage are easy to the industrialization commercialized development, for agricultural sustainable development creates conditions.
Embodiment:
Embodiment 1:
BJNF-1 is seeded on the potato dextrose agar (PDA), 25 ℃ of dark cultivations 3 days, can form a large amount of three-dimensional predation bacterium nets, and begin to produce chlamydospore and a spot of conidium, and the aerial hyphae prosperity, cotton-shaped, bacterium colony is light brown, colony diameter can reach 75mm in 6 days, produced a large amount of chlamydospores, bacterium colony overstrike or Vandyke brown after 10 days.
Embodiment 2:
Dried wheat straw stalk is cut into the long stalk section of 2mm, with wheat stalk section suction and make and drain the water after saturated; Potato is boiled and makes its one-tenth pasty state with suitable quantity of water; Make dried wheat straw stalk, potato, glucose, three's mass ratio is 25: 10: 1, mixes, mixes evenly; 121 ℃ of high pressure steam sterilizations 60 minutes are cooled to room temperature.Make wheat straw bar potato glucose solid medium.
Embodiment 3:
BJNF-1 is seeded on the potato dextrose agar inclined-plane, cultivated 5 days for 25 ℃.Cultivation thalline on the test tube slant is all scraped, be seeded in the 500ml Erlenmeyer flask of filling liquid potato glucose substratum, substratum loading amount 100ml/ bottle places on the shaking table that 25 ℃ of vibrations are dark cultivates 6 days as solid enlarged culturing seed.In mass ratio: solid medium: liquid seeds is 50: 1 a ratio, the enlarged culturing seed is inoculated on the wheat straw bar potato glucose solid medium, and abundant mixing, 28 ℃ of dark cultivations obtained the BJNF-1 solid culture in 7 days.
Embodiment 4:
The BJNF-1 solid culture is natural air drying under normal temperature (25 ℃), from chlamydospore, with sterilized water chlamydospore is carried out gradient dilution with vibrosieve, carries out the chlamydospore counting with counting ware (nematodal accounting ware), and chlamydospore content reaches 10
6Individual every gram dry weight.
Embodiment 5:
After the BJNF-1 solid culture passes through the following natural air drying of normal temperature (25 ℃), drying at room temperature is preserved after six months, 1 year, 2 years, 3 years, get a certain amount of yeast culture thing respectively, add quantitative aseptic water, chlamydospore is washed, draw the chlamydospore suspension behind the gradient dilution with vibrator, coat on the PDA flat board, 25 ℃ of dark cultivations after 3 days, statistics chlamydospore sum and the chlamydospore number of having sprouted calculate the chlamydospore survival rate under stereoscope., drying at room temperature preserves that the chlamydospore survival rate is that the chlamydospore survival rate is that the chlamydospore survival rate is that its chlamydospore survival rate is 15% after 60%, three year after 85%, two year after 98%, one year after six months.
Claims (8)
1. microorganism, it is characterized in that it be a kind of nematode-trapping fungi BJNF-1 that is used for plant nematode and the biological control of intestine of ruminants parasitic nematode every referring to spore (Dactylella sp.), its preservation registration number is: CGMCC No.1051.
2. by the described method for culturing microbes of claim 1, it is characterized in that above-mentioned nematode-trapping fungi separation and Culture by the following method: be added on the nutrient agar that is connected to pedotheque as bait with nematode, 25～28 ℃ of dark cultivations after 5～8 days, to prey on the bacterium net chooses to potato dextrose agar, substratum includes penicillin, Streptomycin sulphate and duomycin and is respectively 20 μ g/ml, 40 μ g/ml, 30 μ g/ml, cultivate this nematode-trapping fungi.
3. by claim 1 or 2 described microorganism preparation methods, it is characterized in that this preparation method carries out according to the following steps:
(1). test tube is cultivated: adopt potato dextrose agar, this bacterium is seeded on the test tube substratum, cultivated 3～5 days for 25～30 ℃.
(2). enlarged culturing: adopt liquid potato glucose substratum, the cultivation bacterium in the above-mentioned test tube is seeded in the Erlenmeyer flask of filling liquid substratum, 25～30 ℃ of vibrations are secretly cultivated 5～6 days as the solid culture seed on shaking table.
(3). solid culture: adopt wheat straw bar potato glucose solid medium, in mass ratio: solid medium: liquid seeds is 50: 1 a ratio, the enlarged culturing seed is inoculated on the solid medium, mixing, 28 ℃ of dark cultivations obtained solid culture in 5～7 days.
4. by the described wheat straw bar of claim 3 potato glucose solid medium, it is characterized in that: dried wheat straw stalk is cut into the long stalk section of 2mm, and the wheat stalk section being absorbed water and make drains the water after saturated; Potato is boiled and makes its one-tenth pasty state with suitable quantity of water; Make dried wheat straw stalk, potato, glucose, three's mass ratio is 25: 10: 1, mixes, mixes evenly; 121 ℃ of high pressure steam sterilizations 30 minutes are cooled to room temperature.
5. by the described solid culture of claim 3, it is characterized in that reaching 106 every gram dry weights through the solid culture chlamydospore content after the normal temperature seasoning.After 3 years, its chlamydospore survival rate is 15% through the process of the solid culture after normal temperature seasoning normal temperature state of nature kept dry.
6. by the described plant nematode of claim 1, it is characterized in that mainly comprising Meloidogyne Meloidogyne, Heterodera Heterodera and Ditylenchus Ditylenchus destructor nematode.
7. by the described intestine of ruminants parasitic nematode of claim 1, it is characterized in that mainly comprising haemonchus (Haemonchus), esophageal orifice nematode (Oesophagostomum), Bunostomum trigonoce phalum (Bunostomum), oersted nematode (Ostertagia), Xia Baite nematode (Chabertia) and sheep net filaria (Dictyocaulus).
8. by the described application of microorganism of claim 1, it is characterized in that nematode-trapping fungi is used to prevent and treat plant nematode and intestine of ruminants parasitic nematode.
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| CN 200410090815 CN1775942A (en) | 2004-11-15 | 2004-11-15 | Predacious roundworm fungus bacterial strain and its preparing method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101831388A (en) * | 2010-05-18 | 2010-09-15 | 华南农业大学 | Nematophagous fungus and preparation method and application thereof |
| CN103952319A (en) * | 2014-04-15 | 2014-07-30 | 西北民族大学 | Nematode-eating fungus separating, purifying and storing culture medium |
| CN104450538A (en) * | 2014-11-26 | 2015-03-25 | 中国农业科学院生物技术研究所 | Culture medium and culture method for Drechmeria coniospora ARSEF 6962 |
| CN105594750A (en) * | 2015-09-14 | 2016-05-25 | 北京晨奥润泽科技股份有限公司 | Microbial insecticide and preparation method thereof |
| CN106173209A (en) * | 2016-07-21 | 2016-12-07 | 蔡葵蒸 | A kind of nematode-destroying fungus powderous preparations and application thereof |
| CN112522115A (en) * | 2020-12-09 | 2021-03-19 | 云南大学 | Application of microbacterium Paraoxidans in inducing Arthrobotrys oligospora to generate predatory organ and method |
-
2004
- 2004-11-15 CN CN 200410090815 patent/CN1775942A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101831388A (en) * | 2010-05-18 | 2010-09-15 | 华南农业大学 | Nematophagous fungus and preparation method and application thereof |
| CN103952319A (en) * | 2014-04-15 | 2014-07-30 | 西北民族大学 | Nematode-eating fungus separating, purifying and storing culture medium |
| CN104450538A (en) * | 2014-11-26 | 2015-03-25 | 中国农业科学院生物技术研究所 | Culture medium and culture method for Drechmeria coniospora ARSEF 6962 |
| CN104450538B (en) * | 2014-11-26 | 2017-07-07 | 中国农业科学院生物技术研究所 | Mould culture medium and cultural method in a kind of Jue Shi plums for circular cone |
| CN105594750A (en) * | 2015-09-14 | 2016-05-25 | 北京晨奥润泽科技股份有限公司 | Microbial insecticide and preparation method thereof |
| CN106173209A (en) * | 2016-07-21 | 2016-12-07 | 蔡葵蒸 | A kind of nematode-destroying fungus powderous preparations and application thereof |
| CN112522115A (en) * | 2020-12-09 | 2021-03-19 | 云南大学 | Application of microbacterium Paraoxidans in inducing Arthrobotrys oligospora to generate predatory organ and method |
| CN112522115B (en) * | 2020-12-09 | 2022-09-02 | 云南大学 | Application of microbacterium Paraoxidans in inducing Arthrobotrys oligospora to generate predatory organ and method |
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