CN1138096A - Strain for controlling cytocyst nematode disease of soybean and its preparation - Google Patents
Strain for controlling cytocyst nematode disease of soybean and its preparation Download PDFInfo
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- CN1138096A CN1138096A CN 95121114 CN95121114A CN1138096A CN 1138096 A CN1138096 A CN 1138096A CN 95121114 CN95121114 CN 95121114 CN 95121114 A CN95121114 A CN 95121114A CN 1138096 A CN1138096 A CN 1138096A
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Abstract
A bacterial strain (CGMCC0241) for preventing and eliminating cystonematodiasis of soybean is disclosed. Its liquid preparation is made up through inoculation of said bacterial strain, three-stage fermentation and addition of compound additive. The parameters for fermentation are 24-30 deg.C, 5-8 of pH value and fermentation stop condition that the number of spores is more than 500,000,000/ml. Its solid preparation may also be made up.
Description
The present invention relates to microorganism field, or rather, relate to and be used to prevent and treat the plant root disease, especially for the bacterial strain and the preparation thereof of control soybean cyst nematode.
Soybean is one of important raise crop of China, and its economic worth is higher.Because cultivated area constantly enlarges, heavily meet batch farming problem and give prominence in recent years, thereby cause the soybean diseases and insect pests onset area to increase, endanger serious day by day.In many disease and pests, with the sick harm maximum of soybean Cyst nematode (Hetero derag1ycines), its onset area accounts for more than 70% of soybean acreage, and is generally with a toll of 10-40%, serious even can cause the soybean total crop failure.For the disease of soybean root, majority is to adopt chemical pesticide and the agent of various chemical seed clothing such as furans pellet, formulation of Duofu mixture and ethyl sulphur phosphorus at present.Though this class medicine has certain preventive effect in earlier stage at soybeans they grow,, cause the severe contamination of environment owing to a large amount of uses of this class chemical pesticide.In addition, because medicine has reduced soil fertility, and germ and insect are developed immunity to drugs in the medium-term and long-term accumulation of soil, reduced the prevention effect of medicine to disease and pest.In view of many disadvantages of chemical pesticide, make that utilizing biological method to prevent and treat the plant nematode disease receives publicity day by day.In the process of biological control, the problem of most critical is to locate and isolates to be had the microorganism strains of enough killing ability and makes the preparation that is suitable for using with this bacterial strain specific disease and pest.Up to now, prevent and treat root knot nematode (Meloidogyne spp.) with Paecilomyces lilacinus (Paccilomyceslilacinus) and done many tests abroad, for example, the existing commodity preparation (trade(brand)name BIOCON) of Philippines is put on market, and the effect that this preparation is prevented and treated its root knot nematode disease by the immersion potato tuber is fine.Yet this preparation is but very undesirable for the effect of control soybean cyst nematode, utilizes dark purple Paecilomyces varioti preparation to prevent and treat the report of soybean cyst nematode so still find no up to now.This be because, for most of plants, its living environment is very complicated, the condition of each side such as microbial inoculum and plant itself, disease and insect, soil interacts.Mostly biocontrol fungicide in the past is that the mode of action and action site are single, thereby prevention effect is limited, especially to being used for the biocontrol fungicide of plant root disease, because the soil fungistasis ubiquity of [be soil suppress fungal spore (also comprising bacterium, actinomycetic propagulum) sprout] has had a strong impact on the disease and insect control that microbial inoculum is applied to big Tanaka.Particularly concerning the soybean of establishing in large scale, do not find a kind of biological control method that can prevent and treat soybean cyst nematode effectively so far yet.
The objective of the invention is to provide a kind of bacterial strain and preparation thereof that is applicable to the control soybean cyst nematode.
The inventor, recorded and on average contains several 67.5 of cyst in every 100g soil in the field piece soil in 4 years of soybean continuous cropping on Meng Jia hilllock, the Hua Nan of Heilongjiang Province county the screening and separating work of various relevant bacterial strains through for many years in 90 years.Detected 128 cysts altogether, parasitic fully accounts for 9.5%.Found that wherein a cyst is very special, the material in the cyst is all cleared up, and fungal spore has been full of whole body cavity, spore is transferred to cultivated breeding on the PDA flat board, through being accredited as the specific strain of Paecilomyces lilacinus.The result that more than 10,000 cyst of kind of soil sample surplus 50 measured in addition, though finding has some bacterial parasites to belong to Paecilomyces lilacinus, but all find to resemble above-mentioned bacterial strains can produce a large amount of spores in the female polypide of soybean Cyst nematode phenomenon, just in cyst, produce mycelia, parasitism or be wrapped on the ovum.Thereby the inventor determines that above-mentioned isolated specific strain is a kind of new bacterial strain.The bacterial strain that obtains is changed in the PDA substratum that contains derosal (from 0.1ppm to 50ppm, increasing concentration gradually), and the result filters out a kind of biocontrol strain that can the derosal of anti-50ppm.After repeatedly changeing generation, the product spore ability of this bacterial strain can descend to some extent, but can make its rejuvenation by the method for uv induction, i.e. 15-20cm place under the 20w ultraviolet lamp, the next day irradiation once, get final product the answer part in the choosing colony behind the about at every turn 15min, general irradiation 3-4 time, carry out monospore and separate, can realize the bacterial strain rejuvenation.When this bacterial strain was cultivated on the PDA substratum, the bacterium colony front was dark violet redness, and the back side is chocolate.After cultivating for two weeks, its colony diameter promptly reaches 79 ± 4mm, produces a large amount of conidiums, and its size is (2.4 ± 1.1) * (2.2 ± 0.3) μ m.This kind bacterium can anti-50ppm derosal, can parasitize in the ovum of soybean Cyst nematode and in the female polypide and can clear up utilization material wherein, finally make its conidium be full of whole female polypide chamber and cause female worm death, very fast quilt is eliminated owing to can't breed thereby make the soybean Cyst nematode.And other bacterial strains in the Paecilomyces lilacinus of being measured are not all found above-mentioned effect.So, the inventor's new bacterial strain that this is special is sent to Beijing China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation, its preservation date is October 23 nineteen ninety-five, and preserving number is CGMCC0241, and its name is called Paecilomyces lilacinus CGMCC0241.In addition,, and add the preparation that composite additive is made this bacterial strain, then be more convenient on large-area soybean planting field, being used for the control of soybean cyst nematode if Paecilomyces lilacinus CGMCC0241 of the present invention is adopted the three grade fermemtation technology of liquid fermenting.
Thereby, the invention provides a kind of bacterial strain that is used to prevent and treat soybean cyst nematode, it is characterized in that it is Paecilomyces lilacinus CGMCC0241.
In addition, the present invention also provides a kind of strain liquid preparation that is used to prevent and treat soybean cyst nematode, and it is to adopt the fungal bacterial strain inoculation, makes by liquid three grade fermemtation technology, it is characterized in that,
The bacterial strain that is used to inoculate is Paecilomyces lilacinus CGMCC0241;
The temperature of liquid fermenting generally is controlled at 24-30 ℃, is preferably 26-28 ℃; PH value generally is controlled at 5-8, preferred PH6-7;
The grasp condition of said final concluding time of three grade fermemtation is that the living spore concentration of the observed bacterial strain liquid of microscopy reaches 5 * 10
8Individual/as more than the ml, preferably to reach 7 * 10
8Individual/more than the ml, and the mycelia dry weight reaches more than 2% (W/V).
Above-mentioned three grade fermemtation technology is meant from the test tube original seed, shake the three grade fermemtation of bottle, seeding tank, fermentor tank through triangle, inoculum size is generally 5-10%, these are prior aries, but concerning Paecilomyces lilacinus of the present invention, temperature during the fermentation, pH value and fermentation time all should be limited in the above-mentioned scope.Fermentation time is generally relevant with the volume of the composition of substratum, leavening temperature, fermentor tank, aeration condition etc.Usually the fermentation time that shakes bottle and seeding tank respectively is 1.5-2.5 days, and the fermentation time of fermentor tank is 2-4 days, and total fermentation time is about a week.
The used substratum of above-mentioned liquid fermenting is provided with strict restriction, it can adopt fungi culture medium general in the prior art, but the inventor finds by the research back, according to the regularity of distribution of soybean nutritional demand and China's soybean main producing region soil mineral element, should suitably add the trace element (as boron, molybdenum etc.) of certain proportion (from several ppm to hundreds of ppm).In substratum, add certain sodium-chlor and calcium salt (0.1-2%), can increase spore production rate significantly and suppress varied bacteria growing.
Therefore, the medium component that the present invention adopts is as follows: medium component content sucrose 2-5% (W/V, down together) NaCl 0.2-2%KNO
30.1-0.8%K
2HPO
40.02-0.2%CaCO
3And/or CaCl
20.1-0.8%FeSO
47H
2O 50-300ppmZnSO
450-300ppmMgSO
47H
2O 200-1000ppmCuSO
42-10ppmNaMoO
42H
2O 2-10ppmH
3BO
45-30ppm water surplus (supplying 100%)
In mentioned component, some elements (chlorine and calcium) have the effect that suppresses the unwanted bacteria growth.Simultaneously, an amount of calcium salt can improve the plant tolerance to diseases to a certain extent.In addition, owing to lack molybdenum and boron in the geographic soil of China's soybean planting, it is very important therefore suitably to replenish these two kinds of trace elements.Other various compositions all are the compositions that helps the breeding of Paecilomyces lilacinus or help soybeans they grow.
On the basis of strain liquid fermentation, the control fermentating liquid PH value, and compound multiple additives is developed into and has the multiple mode of action, the novel biocontrol fungicide of multidigit point-soybean guarantor root fungus agent.At first, fungi growth breeding for the benefit of transfers to slightly acidic condition (PH6.0-6.5) with fermentating liquid PH value.The chitin composition can discharge line material extremely on the one hand after decomposition, can stimulate again simultaneously and strengthen biocontrol microorganisms infecting nematode, but also abundant nutrition is provided can for microorganism, plant, and overcome the restraining effect of soil to fungal inoculant, strengthen the plant tolerance to diseases.After fermentative production finishes, select to add the material (being mainly the crust of animals such as shrimp, crab) that 0.2-0.5% (W/V) is rich in chitin, collagen protein.In addition, some plants have good nematicide effect, are chosen in the garlic juice and/or the hemp juice that add 2-5% (W/V) in the fermented liquid.In addition, in bacterium liquid, add the methylcellulose gum of 0.1-0.3% (w/v), to increase bacterium liquid viscosity and uniformity coefficient, make simultaneously microbial inoculum more firmly dressing on soybean seeds.
Three above-mentioned class additives both can use arbitrary class wherein separately, also can use wherein wantonly two classes or three classes to be used in combination.
In addition, the another kind of preferred version of strain liquid preparation of the present invention is characterised in that, wherein said bacterial strain fermentation liquor is through concentrating under reduced pressure, and then adds in the above-mentioned various additive one or more and carry out composite and make.Wherein said additive comprises the material that is rich in chitin, collagen protein; Garlic juice and/or hemp juice; Methylcellulose gum.The advantage of this scheme is to be convenient to store and transportation.
In addition, the another kind of preferred version of strain liquid preparation of the present invention is characterised in that wherein said bacterial strain fermentation liquor is handled the solid, liquid component respectively, packed through filtering, and makes the preparation that mixing is used when waiting to sow.The advantage of this scheme also is to be convenient to store and transportation.
In addition, the present invention also provides a kind of solid bacterium pearl preparation that is used to prevent and treat soybean cyst nematode, it is characterized in that, it be by aforesaid liquid bacterium pearl preparation and solid filler by (0.5-1.3): 1 (w/w) mixes, dries in the shade and pulverize and make.
Above-mentioned solid filler is preferably mixed by diatomite 70 weight % and the peat composed of rotten mosses 30 weight %.
The using method of liquid bacterial strain preparation of the present invention is as follows: the liquid bacterial agent that adds 100ml by every mu of used soybean seeds amount of sowing (about 5-6kg soybean) mixes, and coating seeds can normally be sowed after waiting to dry.According to different tillage conditions and industrial scale, can also adopt other aftertreatment technologys in addition, it is composite for example liquid bacterial agent to be concentrated the back, or with filtering fermentation liquor, the solid, liquid component is handled packing respectively, and remix is used when waiting to sow.
The using method of solid bacterial strain preparation of the present invention is to sow normally after the pulverizing particulate of said preparation or its shaped particles agent and soybean seeds are mixed again.
Or similar biological farming about with the chemistry farming of prior art compared approximately, the liquid or solid bacterial strain preparation that is used for preventing and treating soybean cyst nematode of the present invention has following advantage: 1, can parasitize in the cyst of nematode, the female polypide and ovum, suppress its growth and breeding, thereby reduce nematode population quantity in the soil significantly; 2, the metabolite that produces in the liquid fermenting process of Paecilomyces lilacinus can press down 2 instar larvaes that kill Cyst nematode; 3, the bacteriostatic action of soil can be overcome effectively, the nematicide ability of Paecilomyces lilacinus can be kept chronically; 4, can grow rhizosphere surely, the dip-dye of antagonism nematode and root-rot bacterium in soybean plant strain; 5, adjustable root section border microenvironment (comprising enzymic activity and microflora) helps soybeans they grow; 6, increase soybean fibrous root amount, and increase root nodule numbers and volume; 7, promote soybeans they grow, increase the TOLERANCE OF SOYBEAN characteristic of disease; 8, can discharge the material that presses down nematicide and germ after the organic additive in the preparation decomposes, promptly act on nematode each stage of the life history, thereby can keep the ability of nematicide and inhibition germ chronically by a plurality of sites, multiple mode.
Further explain the present invention below by specific embodiment.
Embodiment 1
Paecilomyces lilacinus CGMCC 0241 original seed that adopts test tube to cultivate, shake the three grade fermemtation technology cultivation of bottle → seeding tank → fermentor tank again by triangle, inoculum size is 7%, the culture media composition composition that adopts is as follows: medium component content sucrose 3% (W/V, down together) NaCl 1%KNO
30.3%K
2HPO
40.1%CaCO
30.3%FeSO
47H
2O 100ppmZnSO
4100ppmMgSO
47H
2O 500ppmCuSO
45ppmNaMoO
42H
2O 5ppmH
3BO
410ppm water surplus (supplying 100%)
The control leavening temperature is at 26-28 ℃; PH6.5 ± 0.5 I and II fermentation time respectively is decided to be 2 days, and the three grade fermemtation concluding time is decided to be in the fermented liquid liquid and gives birth to spore concentration and reach 6 * 10
8Individual/ml, whole fermentation time is 7 days.The mycelia dry weight that records after the fermentation ends is 2% (W/V).Regulating bacterium liquid pH value is 6.0, the methylcellulose gum of garlic juice and 0.2% (W/V) that adds crab shell powder, 3% (W/V) of 0.25% (W/V) in this fermented liquid, mix the back and mix seed dressing with 3kg soybean seeds (one mu consumption), test 100 mu of ground altogether by every 100ml preparation.In addition, for the purpose of contrasting, mark 10 mu of ground at areal and do not use preparation of the present invention.The result shows that in 100 mu of soybean of using preparation of the present invention, the Cyst nematode disease obviously alleviates, and contrast reduces 65%, and more than 40%, the average yield per mu of soybean exceeds about 20% than the average yield per mu of not using preparation of the present invention to the root rot preventive effect.
Claims (10)
1, a kind of bacterial strain that is used to prevent and treat soybean cyst nematode is characterized in that, it is Paecilomyces lilacinus CGMCC0241.
2, a kind of strain liquid preparation that is used to prevent and treat soybean cyst nematode, it is to adopt the fungal bacterial strain inoculation, makes by liquid three grade fermemtation technology, it is characterized in that,
The bacterial strain that is used to inoculate is Paecilomyces lilacinus CGMCC0241;
The temperature of liquid fermenting generally is controlled to be 24-30 ℃; PH value is 5-8;
The grasp condition of said final concluding time of three grade fermemtation is that the living spore concentration of the observed bacterial strain liquid of microscopy reaches 5 * 10
8Individual/more than the ml, and the mycelia dry weight reaches more than 2% (W/V).
3, strain liquid preparation as claimed in claim 2, it is characterized in that, the composition of the culture media composition that uses in the fermenting process when the preparation said preparation is as follows: medium component content sucrose 2-5% (W/V, down together) NaCl 0.2-2%KNO
30.1-0.8%K
2HPO
40.02-0.2%CaCO
3And/or CaCl
20.1-0.8%FeSO
47H
2O 50-300ppmZnSO
450-300ppmMgSO
47H
2O 200-1000ppmCuSO
42-10ppmNaMoO
42H
2O 2-10ppmH
3BO
45-30ppm water surplus (supplying 100%)
4, strain liquid preparation as claimed in claim 2 is characterized in that, also contains the material that 0.2-0.5% (W/V) is rich in chitin, collagen protein in said preparation.
5, strain liquid preparation as claimed in claim 2 is characterized in that, also contains garlic juice and/or the hemp juice of 2-5% (W/V) in said preparation.
6, strain liquid preparation as claimed in claim 2 is characterized in that, also contains the methylcellulose gum of 0.1-0.3% (w/v) in said preparation.
7, strain liquid preparation as claimed in claim 2 is characterized in that, also contains the material that 0.2-0.5% (W/V) is rich in chitin, collagen protein in said preparation; The methylcellulose gum of the garlic juice of 2-5% (W/V) and/or hemp juice and 0.1-0.3% (w/v).
As strain liquid preparation as described in the claim 2, it is characterized in that 8, wherein said bacterial strain fermentation liquor is through concentrating under reduced pressure, and then add additive described in the claim 7 one or more carry out composite and make.
As strain liquid preparation as described in the claim 2, it is characterized in that 9, wherein said bacterial strain fermentation liquor is handled the solid, liquid component respectively, packed through filtering, and makes the preparation that mixing is used when waiting to sow.
10, a kind of solid bacterial strain preparation that is used to prevent and treat soybean cyst nematode is characterized in that, it be by liquid bacterial agent and solid filler by (0.5-1.3): 1 (W/W) mixes, and dries in the shade and pulverizes and make.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN95121114A CN1058289C (en) | 1995-12-29 | 1995-12-29 | Strain for controlling cytocyst nematode disease of soybean and its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN95121114A CN1058289C (en) | 1995-12-29 | 1995-12-29 | Strain for controlling cytocyst nematode disease of soybean and its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1138096A true CN1138096A (en) | 1996-12-18 |
| CN1058289C CN1058289C (en) | 2000-11-08 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN95121114A Expired - Fee Related CN1058289C (en) | 1995-12-29 | 1995-12-29 | Strain for controlling cytocyst nematode disease of soybean and its preparation |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1317383C (en) * | 2005-03-16 | 2007-05-23 | 云南大学 | Cystic monacrosporium janus prepn with nematocide function and its preparing method and use |
| CN100415888C (en) * | 2005-07-22 | 2008-09-03 | 中国农业科学院植物保护研究所 | Soy bean cyst roundworm specific SCAR label, specific primer and quick PCR detecting method |
| CN101220335B (en) * | 2007-12-28 | 2010-04-14 | 中国烟草总公司郑州烟草研究院 | Bacterial strain for preventing and controlling tobacco black shank and bacterium agent thereof |
| CN101081982B (en) * | 2006-05-29 | 2010-05-12 | 河南农业大学 | Novel vegetable root nematode prevention and cure organic modifying agent |
| CN102286421A (en) * | 2011-09-16 | 2011-12-21 | 中国林业科学研究院森林生态环境与保护研究所 | Liquid fermentation culture method for paecilomyces lilacinus |
| CN102960369A (en) * | 2012-11-22 | 2013-03-13 | 北京市西山试验林场 | Preparation method of granular paecilomyces lilacinus biological nematocide |
| CN104719345A (en) * | 2015-03-31 | 2015-06-24 | 青州伟格生物科技有限公司 | Preparation method of biological preparation for preventing and controlling vegetable nematode disease |
| CN105123771A (en) * | 2015-09-16 | 2015-12-09 | 佛山市盈辉作物科学有限公司 | Pesticide combination |
| CN110771631A (en) * | 2019-12-10 | 2020-02-11 | 云南大学 | Method for preventing and treating tobacco nematode diseases by using compound nematode-killing microorganisms |
Families Citing this family (1)
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| CN108624537B (en) * | 2018-05-29 | 2019-09-03 | 沈阳农业大学 | The microbacterium of one plant of anti-soybean cyst nematode of inducing soybean and its application |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991002051A1 (en) * | 1989-08-03 | 1991-02-21 | The Australian Technological Innovation Corporation | Myconematicide |
| EP0623672A1 (en) * | 1993-05-04 | 1994-11-09 | Research Institute For Plant Protection | New alkaline serine protease of Paecilomyces lilacinus |
-
1995
- 1995-12-29 CN CN95121114A patent/CN1058289C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1317383C (en) * | 2005-03-16 | 2007-05-23 | 云南大学 | Cystic monacrosporium janus prepn with nematocide function and its preparing method and use |
| CN100415888C (en) * | 2005-07-22 | 2008-09-03 | 中国农业科学院植物保护研究所 | Soy bean cyst roundworm specific SCAR label, specific primer and quick PCR detecting method |
| CN101081982B (en) * | 2006-05-29 | 2010-05-12 | 河南农业大学 | Novel vegetable root nematode prevention and cure organic modifying agent |
| CN101220335B (en) * | 2007-12-28 | 2010-04-14 | 中国烟草总公司郑州烟草研究院 | Bacterial strain for preventing and controlling tobacco black shank and bacterium agent thereof |
| CN102286421A (en) * | 2011-09-16 | 2011-12-21 | 中国林业科学研究院森林生态环境与保护研究所 | Liquid fermentation culture method for paecilomyces lilacinus |
| CN102960369A (en) * | 2012-11-22 | 2013-03-13 | 北京市西山试验林场 | Preparation method of granular paecilomyces lilacinus biological nematocide |
| CN102960369B (en) * | 2012-11-22 | 2014-08-20 | 北京市西山试验林场 | Preparation method of granular paecilomyces lilacinus biological nematocide |
| CN104719345A (en) * | 2015-03-31 | 2015-06-24 | 青州伟格生物科技有限公司 | Preparation method of biological preparation for preventing and controlling vegetable nematode disease |
| CN105123771A (en) * | 2015-09-16 | 2015-12-09 | 佛山市盈辉作物科学有限公司 | Pesticide combination |
| CN110771631A (en) * | 2019-12-10 | 2020-02-11 | 云南大学 | Method for preventing and treating tobacco nematode diseases by using compound nematode-killing microorganisms |
| CN110771631B (en) * | 2019-12-10 | 2021-03-26 | 云南大学 | Method for preventing and treating nematode diseases by using composite nematode-killing microorganisms |
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| Publication number | Publication date |
|---|---|
| CN1058289C (en) | 2000-11-08 |
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