CN1476762A - Biological preparation for controlling parasitic nema of plant - Google Patents
Biological preparation for controlling parasitic nema of plant Download PDFInfo
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- CN1476762A CN1476762A CNA031351344A CN03135134A CN1476762A CN 1476762 A CN1476762 A CN 1476762A CN A031351344 A CNA031351344 A CN A031351344A CN 03135134 A CN03135134 A CN 03135134A CN 1476762 A CN1476762 A CN 1476762A
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Abstract
The present invention relates to a biological control preparation for controlling plant parasite nematode. It makes said strain undergo the processes of solid or liquid culture to obtain somatic culture including hypha, conidium and chlamydospore, and mixes said somatic culture with crab shell powder, shrimp shell powder, chitin, active carbon or straw carbon and other auxiliary materials and adsorption carrier together according to a certain ratio, and then make them into powder or microgranules preparation for controlling plant parasite nematode.
Description
Technical field:
The present invention relates to a kind of biological prevention and control agent of preventing and treating plant nematode, belong to biological pesticide technical field.
Background technology:
Plant nematode is the plant disease that generally takes place in a kind of world wide, and only the known kind of root-knot nematode just reaches kind more than 70, and 3000 various plants that cause harm cause tremendous loss every year.Economic crops such as China plant nematode main harm tobacco, flowers, vegetables, cotton, soybean, peanut, pseudo-ginseng, American Ginseng, the loss that causes is very surprising, the whole world annual because of nematode cause with a toll of 1,000 hundred million dollars (Weischer, 1994).The control of nematode at present mainly relies on chemical nematocide, but chemical nematocide mostly is the huge malicious agricultural chemicals of the natural disposition of going out, and to phreatic pollution and residual, makes its use be subjected to certain restriction.The chemical nematocide of present domestic use simultaneously is mainly external imported product, and price is too expensive.Therefore, develop the nematode biological prevention and control agent and seem particularly important.
Occurring in nature, the natural enemy of nematode is a lot, mainly contains: fungi, bacterium, mite, predatism nematode etc.Wherein maximum is fungi, and research at present at most and what become launch also mainly is fungi.Mainly be divided into according to its characteristic that infects nematode:
1, predation fungus: catch nematode by the predacious organs that the nutrient hypha specialization forms;
2, parasitic fungus: by the fungus of the ovum of parasitic nematode, female worm, cyst etc.;
3, toxigenic bacterium thing: the fungus that can produce toxin poisoning nematode.
What become at present that product comes into the market mainly is predation fungus and parasitic fungus.
Summary of the invention:
The objective of the invention is to overcome the deficiency of prior art, a kind of biological prevention and control agent of preventing and treating plant nematode is provided.
The fungi that the present invention adopts is Chinese Verticillium dahliae (Verticillium sinense) GAUZ 412, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Address: China. Beijing. the Zhong Guan-cun; Preservation date: on May 26th, 2003; The numbering CGMCC NO.0933 that preservation is registered on the books.
China Verticillium dahliae GAUZ 412 is the Verticillium dahliae novel species that the inventor found in 1996.Main morphological feature is: nutrient hypha is separated, and branch often expands; Conidium is upright upright, colourless, and long 50-300 μ m produces 1 bottle stalk or 2-6 bottle stalk is verticillate; The long 4.8-12.2 μ of the base portion that expands m, wide 1.5-2.0 μ m are expanded in bottle stalk bottom.Be tapered top wide 0.5-0.7 μ m from the basal orientation top; 10-50 conidium is gathered into head or is chain; Conidium is not separated, and is colourless, rubber fruit shape, and size is 0.8-1 * 0.8-1.2 μ m.Lack dictyochlamydospore.
China Verticillium dahliae (Verticillium sinense) is variation easily, the original strain poor growth.We are separated to different strains from nematode and soil, wherein most characteristic that does not possess we required " obvious stable control efficiency, comprehensive proterties is good, is easy to large-scale production ".After bacterial strain is screened and improves, obtained the unique bacterial strain of a strain of Chinese Verticillium dahliae, called after: GAUZ 412.It has possessed the basic proterties of suitability for industrialized production simultaneously not only to root-knot nematode (Meloidogyne), the good control efficiency of Cyst nematode (Heterodera) tool.
In view of this bacterial strain may be because (spontaneous, physics or chemical) sudden change, protoplast fusion; conversion or other change by biotechnology or are transformed, and the mutant strain of any bacterial strain of this bacterial classification, mutation and derivative are all in protection scope of the present invention.
The present invention prepares according to a conventional method.Be about to Chinese Verticillium dahliae GAUZ 412, by solid or liquid culture, acquisition comprises mycelia, conidium, chlamydosporic thalline culture, after auxiliary materials such as crab shell powder, shrimp shell meal, chitin, active carbon and careless carbon, crop stalk chaff and absorption carrier mix by a certain percentage, through solid fermentation production and liquid fermentation production, make pulvis or fine granule.
1, solid fermentation production
(1) the test tube kind is cultivated
On the test tube culture medium slant, culture medium prescription is to add 5-10 gram sucrose and 5-10 milliliter glycerine in 1000 milliliters of PDA medium, cultivates 6-10 days down, obtains the test tube kind for 22-26 ℃ with the mycelium of GAUZ 412 or spore inoculating.
(2) bacterial classification shaking table enlarged culture (following all be weight percentage)
The inclined-plane seed is inoculated in the triangular flask liquid nutrient medium, and culture medium prescription is that interpolation 10-20 gram molasses, 2-5 restrain ammonium sulfate among 1000 milliliters of PDA, and 2-4 restrains sodium chloride, and 1-3 restrains potassium dihydrogen sulfate, and 2-4 restrains trisodium citrate, and 5-10 restrains sucrose, 5-10 milliliter glycerine.25-30 ℃ of following shaking table cultivated (rotating speed 200-300rpm), 8-15 days.
(3) solid fermentation prepares biological prevention and control agent
Cultured liquid seeds is inserted in the solid culture medium, the prescription of medium is 40%-60% crop stalk chaff, 10-20% soybean cake powder, 20-30% wheat bran, 10%-20% crab shell powder or shrimp shell meal or chitin, cultivate after 20-30 days for 22-26 ℃, 40-45 ℃ of temperature condition made pulvis or fine granule after the oven dry down.
2, liquid fermentation production
(1) the test tube kind is cultivated identical with (1) in the aforementioned solid fermentation production
(2) bacterial classification shaking table enlarged culture is identical with (2) in the aforementioned solid fermentation production
(3) liquid fermentation prepares biologic product
The bacterial classification that enlarges step by step is inoculated in 10% ratio in the fermentation tank of different capabilities, and culture medium prescription is aforementioned solid fermentation (a 2) culture medium prescription, and cultivation temperature 28-32 ℃, incubation time is 144-240 hour, obtains and produces required mycelium and spore.With thalline culture and 5%-10% crab shell powder or shrimp shell meal or chitin, after the 1%-2% active carbon mixes,, make pulvis or fine granule with careless carbon and absorption of crop stalk chaff and mixing.
This biological prevention and control agent can mix use with other nematicide agricultural chemicals, plant growth regulator, but must guarantee mixed medicament the Chinese Verticillium dahliae that the present invention adopts is had no side effect.
Nematode biological prevention and control agent of the present invention is generally used when crop sowing or transplanting.Method is during when sowing and seed are manured into soil jointly, during transplanting this preparation to be applied around the crop rhizosphere.Serious area is taken place nematode can impose in process of crop growth once, and method is that this preparation is applied around the crop rhizosphere, and every mu of each amount of application is the 2-3 kilogram.
The present invention has obviously stable control efficiency, and comprehensive proterties is good, is easy to suitability for industrialized production.
Embodiment:
Be described in further detail the present invention with embodiment below, but content of the present invention is not limited thereto.
Example one: biological prevention and control agent is to the line eggs parasitism
1, preparation test preparation
GAUZ 412 inoculation are cultivated in culture dish, culture medium prescription is to add 6 gram sucrose and 6 milliliters of glycerine in 1000 milliliters of PDA medium, cultivates after 8 days down, with sterile water spore is washed for 25 ℃, count with blood counting chamber, making spore content is 1 * 10
8The spore suspension of/milliliter.
2, test method
Get the sterile petri dish that 5 milliliters of spore suspensions place 5 centimetres of diameters with suction pipe, insert root-knot nematode egg (Meloidogyne) suspension then, 1000 ovum of every ware inoculation.
The preparation of ovum: the root system that will have a large amount of root-knot nematodes shreds and earth mixture, transplants the tomato seedling in 4 ages in week then, changes once numerous about 5 months.From the pot culture tomato seedling, take out the root system of having inoculated root-knot nematode during use, washing, put into wide-mouth bottle after the chopping, add 1% clorox, vibrate and pour in the double deck screen (going up 250 microns of mesh sizes, 38 microns of following mesh sizes) after 3 minutes into, wash repeatedly all to be flushed to down and sieve up to all ovum, make line eggs suspension after the collection, with nematodal accounting plate counting.
Test repeats 3 times.Contrast adds 5 ml sterile waters.Cultivate down for 25 ℃, each is handled culture dish obturage, pollute and the evaporation of bacterium liquid to reduce with sealing film.Observe nematode hatching situation after 15 days, hatched promptly that explanation is not suppressed, hatching does not illustrate promptly that its ovum has been suppressed and has failed and sprouts, and calculates the ovum inhibiting rate then.
3, result of the test
Table 1 biological prevention and control agent is to the inhibiting rate of root line eggs hatching
| Handle | The nematode number (bar) that hatches | ||||
| Handle 1 | Handle 2 | Handle 3 | On average | Inhibiting rate/% | |
| Biological prevention and control agent | 342 | 318 | ?351 | ?337 | 62.1％ |
| Blank | 892 | 872 | ?906 | ?890 | |
The result shows: microbial inoculum has good inhibitory effect to the ovum hatching of root-knot nematode, and inhibiting rate reaches 62.2%.Microbial inoculum of the present invention is by suppressing the purpose that the ovum hatching reaches the control nematode.
Example two: submerged fermentation prepares biological prevention and control agent and to the test of pesticide effectiveness of root-knot nematode
1, submerged fermentation prepares biological prevention and control agent
(1) with the mycelia of GAUZ 412 or spore inoculating on the test tube culture medium slant, culture medium prescription is to add 6 gram sucrose and 6 glycerine in 1000 milliliters of PDA medium, cultivates down for 25 ℃ and obtains the test tube kind in 8 days.
(2) the inclined-plane seed is inoculated in triangular flask (500 milliliters) liquid nutrient medium, culture medium prescription is to add 12 gram molasses, 3 gram ammonium sulfate, 4 gram sodium chloride, 1 gram potassium dihydrogen sulfate, 2 gram trisodium citrates, 8 gram sucrose, 8 milliliters of glycerine among 1000 milliliters of PDA.Under 26 ℃, shaking table (rotating speed 250rpm) was cultivated 10 days.
(3) the triangular flask liquid spawn that ferments is inserted in 1 ton of fermentation tank, culture medium prescription is identical with aforementioned (2), 30 ℃, cultivate and insert 10 tons of fermentation cylinder for fermentation after 48 hours after 190 hours, emit and 5% crab shell powder or shrimp shell meal or chitin, after 1% active carbon mixes,, make fine granule after 42 ℃ of oven dry with careless carbon and absorption of crop stalk chaff and mixing.
2, test method
(1) test with medicament and processing
2 kilograms/mu of A, biological prevention and control agents, the disposable shoot root border that imposes on when flue-cured tobacco transplanting;
B, A Bading dilute 1000 times of liquid, the disposable shoot root portion that imposes on when flue-cured tobacco transplanting;
C, blank apply clear water.
(2) test method
For studying thing is the big gold dollar of susceptible tobacco bred safflower, and test site is Gong Jiaying village, Ma Jie township, Yiliang County, Kunming, Yunnan Province, and the very serious plot of root-knot nematode (Meloidogyne) takes place over the years, and soil is red soil, middle fertility.
The sub-district is the single file district, and 33 square metres of areas are planted cigarette 50 strains, and 3 repetitions are established in every processing, randomized arrangement, and totally 9 sub-districts are established guard row all around, by high-quality cigarette cultivation management measure standardized management.
(3) investigation method
After gathering last a slice tobacco leaf, flue-cured tobacco carries out the incidence investigation immediately, every sub-district is got the 10 strains investigation of uprooting at random, after root turns over and digs out, the water fine laundering removes silt, observe the pieces of an egg situation with lens examination root knot and attention, by tobacco system root knot nematode disease grade scale (Y/cT40-1996) classification record, calculate diseased plant rate, root knot percentage, the downright bad percentage of root, disease index, control efficiency respectively.
List and gather and toast in each sub-district, different disposal classification is respectively weighed and calculated output, the output value, average price, first-class cigarette and better-than-average cigarette ratio.
4, experimental result
Table 2 biological prevention and control agent is to the control rate of root-knot nematode
| Handle | Incidence | ||||
| Diseased plant rate/% | Root knot percentage/% | Downright bad percentage/the % of root | Disease index | Control efficiency/% | |
| Biological prevention and control agent | 15.2 | ?10 | ?2 | ?16.7 | ?70.5 |
| A Bading | 40 | ?25 | ?10 | ?40 | ?29.5 |
| Blank | 56.7 | ?40 | ?15 | ?56.7 | |
The economic characters index of the different chemicals treatment cigarette of table 3 strain
| Handle | The economic characters index | ||||
| Output kg/m 2 | The output value/unit | Average price/unit (kg) | First-class cigarette ratio/% | Better-than-average cigarette ratio/% | |
| Biological prevention and control agent | 14.15 | 116.3 | ?8.21 | ?41 | ?76.3 |
| A Bading | 10.25 | 83.6 | ?8.15 | ?30.7 | ?69.3 |
| Blank | 9.75 | 66.3 | ?6.8 | ?28.2 | ?65.1 |
The result shows: the diseased plant rate/% of biological prevention and control agent, root knot percentage/%, the downright bad percentage/% of root, disease index are respectively than A Bading low 24.8%, 15%, 8%, 23.3, than blank low 41.5%, 30%, 13%, 40, average preventive effect is than chemical pesticide A Badinggao 41%, shown the good preventive effect of biological prevention and control agent, possessed and produced the primary condition that goes up large-scale popularization.
The economic characters index analysis shows, each economic characters index of biological prevention and control agent is all high than chemical pesticide and contrast, its output, the output value, average price, first-class cigarette ratio and better-than-average cigarette ratio be respectively than chemical pesticide A Badinggao 3.9kg, 32.7 yuan, 0.06 yuan, 10.3%, 7%, and comparison is according to having improved 4.4kg, 50 yuan, 1.41 yuan, 12.8%, 11.2% respectively.
Field control effectiveness test shows that the biological prevention and control agent that the present invention produces has good control efficiency to root-knot nematode, and its preventive effect surpasses chemical pesticide A Bading, can promote the use of on producing.
Example three: solid fermentation prepares biological prevention and control agent
Adopt previous examples two (1) preparation solid test tube slant bacterial classifications, example two (2) preparation triangular flask liquid spawns, cultured triangular flask liquid spawn is inserted in the solid culture medium, the prescription of solid culture medium is 15% soybean cake powder, 20% wheat bran, 55% crop stalk chaff, 10% crab shell powder or shrimp shell meal or chitin, cultivate after 20 days for 25 ℃, 42 ℃ of temperature condition are made pulvis or fine granule after the oven dry down.
Claims (1)
1. biological prevention and control agent of preventing and treating plant nematode, prepare according to a conventional method by producing bacterial strain and auxiliary material, the production bacterial strain that it is characterized in that this biological prevention and control agent is Chinese Verticillium dahliae Verticillium sinense GAUZ 412, this bacterial strain is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on May 26th, 2003, and preserving number is CGMCC NO.0933.
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| CN 03135134 CN1206922C (en) | 2003-06-03 | 2003-06-03 | Biological preparation for controlling parasitic nema of plant |
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| CN 03135134 CN1206922C (en) | 2003-06-03 | 2003-06-03 | Biological preparation for controlling parasitic nema of plant |
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| CN1206922C CN1206922C (en) | 2005-06-22 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1311750C (en) * | 2006-02-14 | 2007-04-25 | 云南大学 | Active-fermented broth and use thereof |
| CN100401900C (en) * | 2005-03-16 | 2008-07-16 | 云南大学 | Wireworm-killing biologic bacterial agent, its preparation and application |
| CN104004663A (en) * | 2014-05-27 | 2014-08-27 | 内蒙古农业大学 | Preparation method of Duddingtonia flagrans biological freeze-drying preparation |
| CN104928192A (en) * | 2015-06-26 | 2015-09-23 | 云南大学 | Trichoderma viride strain and application thereof |
-
2003
- 2003-06-03 CN CN 03135134 patent/CN1206922C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100401900C (en) * | 2005-03-16 | 2008-07-16 | 云南大学 | Wireworm-killing biologic bacterial agent, its preparation and application |
| CN1311750C (en) * | 2006-02-14 | 2007-04-25 | 云南大学 | Active-fermented broth and use thereof |
| CN104004663A (en) * | 2014-05-27 | 2014-08-27 | 内蒙古农业大学 | Preparation method of Duddingtonia flagrans biological freeze-drying preparation |
| CN104928192A (en) * | 2015-06-26 | 2015-09-23 | 云南大学 | Trichoderma viride strain and application thereof |
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| Publication number | Publication date |
|---|---|
| CN1206922C (en) | 2005-06-22 |
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