CN106701581B - A kind of thickness spore Verticillium dahliae agent and its protective agent and preparation method - Google Patents
A kind of thickness spore Verticillium dahliae agent and its protective agent and preparation method Download PDFInfo
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Abstract
The present invention provides a kind of thick spore Verticillium dahliae agent and its protective agent and preparation methods.Microbial inoculum of the invention contains protective agent; the protective agent contains the raw material of following parts by weight; the mycelia of thick spore Verticillium dahliae is uniformly mixed with protective agent of the invention and is dried to obtain thick spore Verticillium dahliae agent after passing through the quick pelletizer granulation of bimodulus by 30-50 parts of zeolite powder, 15-25 parts of humic acid, 25-45 parts of cornstarch, 0.2-2 parts of sucrose, 0.1-1.5 parts of sodium chloride, 0.1-1.2 parts of Tween 80,0.01 part of chloramphenicol.Protective agent of the invention can effectively extend the validity period of thick spore Verticillium dahliae, and thickness spore Verticillium dahliae agent stability obtained is good, and long shelf-life is easy to maintain, and effect is not easy to lose, and can be widely applied to the root knot nematode disease of crops, promote plant growth.
Description
Technical field
The present invention relates to a kind of microbial bacterial agents, and in particular to a kind of thick spore Verticillium dahliae agent and its guarantor for biological control
Protect agent and preparation method.
Background technique
Thick spore Verticillium dahliae also known as thickness wall spore Pu Keniya bacterium (Pochonia chlamydosporia), the Pseudomonas Yu Banzhi
Bacterium subphylum (Deuteromycotina), Hyphomycetes (Hyphomycetes), hyphomycetales (Moniliales) (Moniliales), silk
Spore section (Moniliaceae) (Monilaccae), Pukeniya genus (Pochonia).
Root-knot nematode is the pathogen for causing crop root knot nematode disease, and host range is very extensive, be popular in the torrid zone and
Massive losses are all carried out to the economy-zone of world crops every year in subtropical zone.Currently, prevention and treatment root knot nematode disease method have it is more
Kind, wherein influence of the method for biological control to people and animals and environment is smaller, is the control safest mode of nematode, prevents and treats line at present
The bacterium that worm is most widely used is Paecilomyces lilacinus and thick spore Verticillium dahliae.
About thick spore Verticillium dahliae, research direction is mainly its biocontrol effect, fermentation condition optimization, the parasitism with worm's ovum at present
Mechanism study etc..But also it is commercialized in someone, and obtains good effect.Wang Laifa etc. is by Paecilomyces lilacinus solid
Microbial inoculum and thick spore Verticillium dahliae microbial inoculum are mixed and made into composite bacteria agent, can obtain indoors, in cell and field experiment preferable anti-
Control effect, the mutual not antagonism of the two, and plant growth can be promoted.Zhuan Zhanxing etc. prevents and treats root knot nematode in tobacco using thick spore Verticillium dahliae,
Prevention and treatment efficiency is 70% or more in field trials, and can promote crop yield.But thick spore Verticillium dahliae will be commercialized, also
Needing to improve its validity period, Shi Yan's studies have shown that is made into wettable powder under the conditions of 4 DEG C for thick spore Verticillium dahliae conidia powder,
After 90 days, effective viable bacteria has dropped 30%, and preservation then has dropped 75.38% under normal temperature conditions.Thick spore Verticillium dahliae is not easy to protect
It deposits and brings difficulty to carrying out biological prevention and control using it, therefore research and develop a kind of more efficiently protective agent to make thick spore Verticillium dahliae
Validity period it is longer just very significant.
Summary of the invention
The purpose of the present invention is overcoming the prior art, thick spore Verticillium dahliae storage life can effectively be extended by providing one kind
Protective agent and long shelf-life thick spore Verticillium dahliae microbial inoculum and preparation method thereof.
Present invention firstly provides a kind of composition, the raw material containing following parts by weight: 30-50 parts of zeolite powder, humic acid 15-
25 parts, 25-45 parts of cornstarch, 0.2-2 parts of sucrose, 0.1-1.5 parts of sodium chloride, 0.1-1.2 parts of Tween 80, chloramphenicol 0.01
Part.
Preferably, composition of the invention contains the raw material of following parts by weight: 35-45 parts of zeolite powder, humic acid 18-23
Part, 30-40 parts of cornstarch, 0.5-1.5 parts of sucrose, 0.4-1.2 parts of sodium chloride, 0.4-1 parts of Tween 80,0.01 part of chloramphenicol.
It is highly preferred that composition of the invention contains the raw material of following parts by weight: 42 parts of zeolite powder, 20 parts of humic acid, jade
36 parts of rice starch, 1.2 parts of sucrose, 1 part of sodium chloride, 0.5 part of Tween 80,0.01 part of chloramphenicol.
The present invention provides above-mentioned compositions to prepare the application in microbial inoculum.
The present invention provides the microbial inoculums containing above-mentioned composition.
Preferably, the microbial inoculum is thick spore Verticillium dahliae (Pochonia chlamydosporia) microbial inoculum.
It is described thickness spore Verticillium dahliae (Pochonia chlamydosporia) microbial inoculum be by thick spore Verticillium dahliae with it is of the present invention
Composition be uniformly mixed according to mass ratio 2:8, adjust moisture to 15%-20%, dried after granulation to 8% or less moisture i.e.
?.Inventor repeatedly the study found that moisture lower than 15% be granulated granulating rate it is bad, be not easy to be granulated greater than 20%, thus select
Moisture is 15%-20%.
Preferably, microbial inoculum of the invention is graininess.The present invention has found that microbial inoculum dosage form of the invention is by numerous studies
Granular pattern is conducive to the extension of shelf-life, other dosage forms are not so good as corpuscular long shelf-life.
The present invention provides a kind of systems of thick spore Verticillium dahliae (Pochonia chlamydosporia) microbial inoculum of long shelf-life
Preparation Method, by the mycelia of thick spore Verticillium dahliae (Pochonia chlamydosporia) and composition of the present invention according to matter
Amount is uniformly mixed than 2:8, is adjusted moisture to 15%-20%, is dried to 8% or less moisture after granulation to obtain the final product.
Wherein, the mycelia of thick spore Verticillium dahliae (Pochonia chlamydosporia) is that thick spore Verticillium dahliae sends out by liquid
Ferment is collected after centrifugation mycelia and obtains.
Thick spore Verticillium dahliae liquid fermentation medium is potato culture medium (PS culture medium), and cultivation temperature is 25-28 DEG C, revolving speed
For 120r/min, ventilatory capacity 2.5kg, incubation time is 5-7 days, and the machinery that mycelia is collected by centrifugation is disc centrifuge, is turned
Speed is 6000r/min.The mycelia of acquisition is mixed with the present composition, adjusts moisture to 15%-20%, bimodulus is quickly granulated
Machine is granulated, dry under the conditions of 30 DEG C after granulation, it is dry to moisture below 8% to obtain the final product.
Further, the application the present invention provides above-mentioned microbial inoculum in prevention and treatment crop nematodiasis.
Further, the present invention provides above-mentioned microbial inoculums to promote the application in plant growth
The beneficial effects of the invention are that:
1, composition provided by the invention can be used for the protective agent of thick spore Verticillium dahliae agent, can enhance the stability of microbial inoculum, prolong
The validity period of long microbial inoculum, according to company standard, microbial inoculum content >=200,000,000 cfu/g are qualified products, and this product validity period is greater than 1 year.
2, bacterial preparation process of the invention is simple to operation, at low cost.
Detailed description of the invention
Fig. 1 is that the thick spore Verticillium dahliae microbial inoculum content of embodiment 6 changes with time.
Fig. 2 is that the thick spore Verticillium dahliae microbial inoculum content of embodiment 7 changes with time.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Without departing substantially from spirit of that invention
In the case where essence, to modifications or substitutions made by the method for the present invention, step or condition, all belong to the scope of the present invention.
Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art；
Unless otherwise specified, agents useful for same is commercially available in embodiment.
The thick spore Verticillium dahliae that the present embodiment uses is purchased from Chinese agriculture Microbiological Culture Collection administrative center, deposit number
ACCC30604。
The preparation (1) of the thick spore Verticillium dahliae agent of embodiment 1
Thick spore Verticillium dahliae liquid fermentation medium is potato culture medium (PS culture medium), and cultivation temperature is 25-28 DEG C, revolving speed
For 120r/min, ventilatory capacity 2.5kg, incubation time is 7 days.With disc centrifuge by fermentation liquid centrifugal treating, revolving speed is
6000r/min.The mycelia of acquisition is mixed with protective agent, wherein zeolite powder 42kg, humic acid 20kg, cornstarch 36kg, sugarcane
Sugared 1.2kg, sodium chloride 1kg, Tween 80 0.5kg, chloramphenicol 0.01kg, thick spore Verticillium dahliae mycelia 25.18kg.Adjust moisture to
20% or so, the quick pelletizer of bimodulus is granulated, dry under the conditions of 30 DEG C after granulation, it is dry to moisture below 8% to obtain the final product.
The preparation (2) of the thick spore Verticillium dahliae agent of embodiment 2
Thick spore Verticillium dahliae preparation by liquid fermentation culture medium is potato culture medium (PS culture medium), and cultivation temperature is 25-28 DEG C,
Revolving speed is 120r/min, and ventilatory capacity 2.5kg, incubation time is 5 days.With disc centrifuge by fermentation liquid centrifugal treating, turn
Speed is 6000r/min.The mycelia of acquisition is mixed with protective agent, wherein zeolite powder 30kg, humic acid 15kg, cornstarch
25kg, sucrose 0.2kg, sodium chloride 0.1kg, Tween 80 0.1kg, chloramphenicol 0.01kg, thick spore Verticillium dahliae mycelia 17.60kg.It adjusts
To 20% or so, the quick pelletizer of bimodulus is granulated moisture content, dry under the conditions of 30 DEG C after granulation, it is dry to moisture 8% with
Down to obtain the final product.
The preparation (3) of the thick spore Verticillium dahliae agent of embodiment 3
Thick spore Verticillium dahliae preparation by liquid fermentation culture medium is potato culture medium (PS culture medium), and cultivation temperature is 25-28 DEG C,
Revolving speed is 120r/min, and ventilatory capacity 2.5kg, incubation time is 6 days.With disc centrifuge by fermentation liquid centrifugal treating, turn
Speed is 6000r/min.The mycelia of acquisition is mixed with protective agent, wherein zeolite powder 50kg, humic acid 25kg, cornstarch
45kg, sucrose 2kg, sodium chloride 1.5kg, Tween 80 1.2kg, chloramphenicol 0.01kg, thick spore Verticillium dahliae mycelia 31.18kg.It adjusts
Moisture is to 20% or so, and the quick pelletizer of bimodulus is granulated, and dry under the conditions of 30 DEG C after granulation, drying is to moisture below 8%
To obtain the final product.
The preparation (4) of the thick spore Verticillium dahliae agent of embodiment 4
Thick spore Verticillium dahliae preparation by liquid fermentation culture medium is potato culture medium (PS culture medium), and cultivation temperature is 25-28 DEG C,
Revolving speed is 120r/min, and ventilatory capacity 2.5kg, incubation time is 5 days.With disc centrifuge by fermentation liquid centrifugal treating, turn
Speed is 6000r/min.The mycelia of acquisition is mixed with protective agent, wherein zeolite powder 35kg, humic acid 18kg, cornstarch
30kg, sucrose 0.5kg, sodium chloride 0.4kg, Tween 80 0.4kg, chloramphenicol 0.01kg, thick spore Verticillium dahliae mycelia 21.08kg.It adjusts
To 20% or so, the quick pelletizer of bimodulus is granulated moisture content, dry under the conditions of 30 DEG C after granulation, it is dry to moisture 8% with
Down to obtain the final product.
The preparation (5) of the thick spore Verticillium dahliae agent of embodiment 5
Thick spore Verticillium dahliae preparation by liquid fermentation culture medium is potato culture medium (PS culture medium), and cultivation temperature is 25-28 DEG C,
Revolving speed is 120r/min, and ventilatory capacity 2.5kg, incubation time is 7 days.With disc centrifuge by fermentation liquid centrifugal treating, turn
Speed is 6000r/min.The mycelia of acquisition is mixed with protective agent, wherein zeolite powder 45kg, humic acid 23kg, cornstarch
40kg, sucrose 1.5kg, sodium chloride 1.2kg, Tween 80 1kg, chloramphenicol 0.01kg, thick spore Verticillium dahliae mycelia 27.93kg.It adjusts
Moisture is to 20% or so, and the quick pelletizer of bimodulus is granulated, and dry under the conditions of 30 DEG C after granulation, drying is to moisture below 8%
To obtain the final product.
The thick spore Verticillium dahliae agent preservation effect test of embodiment 6
Thickness spore Verticillium dahliae agent made from embodiment 1 is saved under the conditions of room temperature and 4 DEG C DEG C respectively, measured it every 15 days
Viable count, measuring method are blood bead tally counting method.Its result is as shown in Figure 1.After preservation in 120 days, detection hair
Thick spore Verticillium dahliae viable count loses 11.26% under the conditions of present 4 DEG C, thick spore Verticillium dahliae viable count loss under normal temperature condition
31.92%.Validity period (the document entitled " thick wall spore Pu Keniya bacterium of the effect better than the wettable powder of the beautiful proposition of stone
PC152 bacterial strain biology and preparation process research ", wettable powder agent prescription are as follows: wetting agent Tween-80 5%, dispersing agent
PEG6000 5%, uv-protector 0.9%, carrier diatomaceous earth 45%, with conidia powder 100% is complemented to.4 DEG C are stored 90 days, damage
23.08% is lost, room temperature storage 90 days, loss was 75.38%).
Above-mentioned comparative test is carried out using microbial inoculum made from 2-5 of the embodiment of the present invention, it is as a result similar with 1 result of embodiment.
The thick spore Verticillium dahliae agent stability test of 7 present invention of embodiment
Thickness spore Verticillium dahliae agent made from embodiment 1 is saved under the conditions of room temperature and 4 DEG C respectively, measured its work every 15 days
Bacterium number, measuring method are blood bead tally counting method.Still there are also 2.15 hundred million cfu/g after preservation in 375 days, according to enterprise
Industry standard (living bacteria count >=200,000,000 cfu/g, standard are higher than living bacteria count >=0.5 hundred million cfu/g of national regulation), this product
Validity period can reach 1 year, although as a result as shown in Figure 2, above having used general explanation, specific embodiment and test,
The present invention is described in detail, but on the basis of the present invention, can make some modifications or improvements to it, this is to this field skill
It is obvious for art personnel.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention,
Belong to the scope of protection of present invention.
Above embodiment be only preferred embodiments of the present invention will be described, not to the scope of the present invention into
Row limits, and without departing from the spirit of the design of the present invention, this field ordinary engineering and technical personnel is to technical side of the invention
The all variations and modifications that case is made, should fall within the scope of protection determined by the claims of the present invention.
Claims (9)
1. a kind of protective agent for being used to prepare thick spore Verticillium dahliae microbial inoculum, which is characterized in that be made of raw material from the following weight: boiling
30-50 parts of mountain flour, 15-25 parts of humic acid, 25-45 parts of cornstarch, 0.2-2 parts of sucrose, 0.1-1.5 parts of sodium chloride, Tween 80
0.1-1.2 parts, 0.01 part of chloramphenicol.
2. being used to prepare the protective agent of thick spore Verticillium dahliae microbial inoculum as described in claim 1, which is characterized in that by following parts by weight
Raw material be made: 35-45 parts of zeolite powder, 18-23 parts of humic acid, 30-40 parts of cornstarch, 0.5-1.5 parts of sucrose, sodium chloride
0.4-1.2 parts, 0.4-1 parts of Tween 80,0.01 part of chloramphenicol.
3. being used to prepare the protective agent of thick spore Verticillium dahliae microbial inoculum as claimed in claim 2, which is characterized in that by following parts by weight
Raw material be made: 42 parts of zeolite powder, 20 parts of humic acid, 36 parts of cornstarch, 1.2 parts of sucrose, 1 part of sodium chloride, Tween 80 0.5
Part, 0.01 part of chloramphenicol.
4. any protective agent of claim 1-3 is preparing thick spore Verticillium dahliae (Pochonia chlamydosporia) bacterium
Application in agent.
5. containing any protectant thick spore Verticillium dahliae microbial inoculum of claim 1-3.
6. thickness spore Verticillium dahliae microbial inoculum as claimed in claim 5, which is characterized in that it is graininess.
7. thickness spore Verticillium dahliae microbial inoculum as claimed in claim 6, which is characterized in that by thick spore Verticillium dahliae (Pochonia
Chlamydosporia) be uniformly mixed with any protective agent of claim 1-3 according to mass ratio 2:8, adjusting moisture to
15%-20% is dried to 8% or less moisture after granulation to obtain the final product.
8. application of any thick spore Verticillium dahliae microbial inoculum of claim 5-7 in prevention and treatment crop nematodiasis.
9. any thick spore Verticillium dahliae microbial inoculum of claim 5-7 is promoting the application in plant growth.
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| CN108013031A (en) * | 2017-11-23 | 2018-05-11 | 云南微态源生物科技有限公司 | A kind of thickness wall spore Pu Keniya bacterium coating method granules and preparation method thereof |
| CN110214793A (en) * | 2019-04-26 | 2019-09-10 | 云南微态源生物科技有限公司 | Thick wall cell verticil branch bacterium biological pesticide novel form and preparation method thereof |
| CN111362760B (en) * | 2020-03-09 | 2022-05-24 | 沃博特生物科技有限公司 | Compound microbial agent-coated biological compound fertilizer and preparation method thereof |
| CN112063536B (en) * | 2020-09-04 | 2022-07-05 | 青岛和协生物科技有限公司 | Pochonia chlamydosporia for preventing and treating root knot nematode disease, composite powder and application |
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Address after: 410000 Changsha, Hunan, China National hi tech Industrial Development Zone, 27 yuan Road, Yu Yuan Pioneer Building 1506, 1507 Applicant after: Taigu ecological science and technology group Limited by Share Ltd Address before: 410205 floor 14, A, Lulu business center, No. 199 Lulong Road, Changsha high tech Zone, Hunan. Applicant before: Hunan Taigu Biotechnology Co., Ltd. |
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| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210331 Address after: Room 310-3, headquarters building, Changsha Zhongdian Software Park Co., Ltd., No. 39, Jianshan Road, Changsha hi tech Development Zone, Changsha, Hunan 410000 Patentee after: Hunan Huigu Agricultural Ecology Research Institute Co.,Ltd. Address before: 410000 Changsha, Hunan, China National hi tech Industrial Development Zone, 27 yuan Road, Yu Yuan Pioneer Building 1506, 1507 Patentee before: TAIGU BIOTECHNOLOGY GROUP Co.,Ltd. |