CN109797110A - A method of acquisition beauveria bassiana is trapped using cylas formicarius - Google Patents
A method of acquisition beauveria bassiana is trapped using cylas formicarius Download PDFInfo
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- CN109797110A CN109797110A CN201910246252.5A CN201910246252A CN109797110A CN 109797110 A CN109797110 A CN 109797110A CN 201910246252 A CN201910246252 A CN 201910246252A CN 109797110 A CN109797110 A CN 109797110A
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Abstract
The method provided by the invention for trapping acquisition beauveria bassiana using cylas formicarius includes: first to carry out cylas formicarius raising and collecting soil sample；The cylas formicarius adult of raising is placed on the pedotheque of acquisition again and is cultivated, realizes and infection processing is trapped to cylas formicarius, beauveria bassiana mycelia and conidium occur on cylas formicarius polypide；By on cylas formicarius polypide beauveria bassiana mycelia and conidium separated from cylas formicarius polypide, and will separation obtain beauveria bassiana mycelia and conidium be transferred in PDA culture medium the beauveria bassiana cultivated, purified.Infection is trapped using cylas formicarius, it overcomes and is easy contaminated defect when directly being separated from the polypide of falling ill that field acquires, acquisition beauveria bassiana is trapped using cylas formicarius, it overcomes polypide of falling ill and acquires the defect big by seasonal effect, therefore technical solution provided by the invention is a kind of efficient, quick beauveria bassiana acquisition method.
Description
Technical field
The application belongs to microorganisms technical field, specifically, being related to a kind of trap using cylas formicarius acquires ball spore
The method of muscardine.
Background technique
Beauveria bassiana (Beauveria bassiana) belongs to Ascomycota, Hypocreales, cordyceps sinensis Cordycepps, Beauveria
A kind of insect pathogenic fungus, bacterium distribution is wide, and pathogenicity is strong, easily culture and nontoxic to people and environment, is not likely to produce anti-
The features such as pharmacological property is the important content in integrated pest control Study on Measures and screening for a long time.The strain is as one at present
The important biocontrol fungi of kind has been used for a variety of evils such as prevention and treatment coleoptera, Semiptera, Homoptera, Lepidoptera, Diptera and Hymenoptera
Worm.Beauveria bassiana is as a kind of important insect pathogenic fungus, because selectivity is high, host range is wide, safety is good, to life for it
State systematic influence is small, be not likely to produce drug resistance, be readily produced, control efficiency is lasting and stablize many advantages, such as, acquisition separation
Concern of the method by people.
Currently, the acquisition separation of beauveria bassiana is usually by collecting the susceptible bombys batryticatus of nature, then moisturizing indoors is trained
It is separated after supporting, or acquisition is trapped using greater wax moth.There are some problems, examples for the acquisition separation method of above-mentioned beauveria bassiana
Such as, be easy when being separated from polypide of falling ill it is contaminated, need to purify and cultivate repeatedly in addition some because pollution is too strong and
Make to separate unsuccessful.Meanwhile when acquiring beauveria bassiana, find the heavy workload of host insect bombys batryticatus, spend the time more and by
Seasonal effect, it is main that summer and autumn occurs since the seasonality that the host insect of beauveria bassiana occurs is very strong, and in spring
Host insect many places are in Wintering Period when lower with winter temperature, along with this season environment temperature is lower, because of then beauveria bassiana
Infection rate it is lower.Therefore, in acquisition and separation, due to being influenced by host insect, i.e., only working as beauveria bassiana
Host insect occur season be possible to by beauveria bassiana infect chance, beauveria bassiana host insect also
Nonevent season, you can't get infected polypides, to affect acquisition and separation.It is adopted in addition, being trapped with greater wax moth
Collection, although infection beauveria bassiana can also be trapped, thus be separately cultured, however since greater wax moth needs to be carried out with honeycomb spleen
Raising, thus steamed bun higher cost, rearing conditions are complex, are not easy to operate.
Summary of the invention
In view of the above drawbacks of the prior art, the application, which provides, a kind of traps acquisition beauveria bassiana using cylas formicarius
Method.
The method provided by the invention for trapping acquisition beauveria bassiana using cylas formicarius, comprising the following steps:
Step 1: cylas formicarius is raised: cylas formicarius being placed in transparent plastic illiciumverum prismatic bottle, is to raise with sweet potato
Material, is raised in 25 ± 1 DEG C of temperature environment indoors；
Step 2: collecting soil sample: selection soil sampling point acquires soil, soil in the plough horizon of sampled point
Acquisition acquires 0.5kg soil with a thickness of 5-7mm, each sampled point, the soil acquired in every 5 sampled points is mixed, as soil
Earth sample；
Step 3: carrying out trapping infection processing to cylas formicarius: the cylas formicarius adult raised through step 1 is set
In on the pedotheque acquired through step 2, cultivated 5-7 days under the conditions of 25+1 DEG C, 16L:8D；
Step 4: production PDA culture medium: in every 1000mL water be added potato 200g, glucose 20g, agar 15~
20g is uniformly mixed；
Step 5: the separation of beauveria bassiana: the ball spore on the cylas formicarius polypide obtained through step 3 culture is white
Stiff bacterium mycelia and conidium separate from cylas formicarius polypide；
Step 6: the purifying culture of beauveria bassiana: by the beauveria bassiana mycelia obtained through step 5 and conidium
It is transferred to and in PDA culture medium made from step 4, is cultivated 3-4 days in 25+2 DEG C of temperature environment, then step 6 is repeated to grasp
Make 2-3 times, the beauveria bassiana purified.
The method as described above for trapping acquisition beauveria bassiana using cylas formicarius, it is preferable that after step 6,
Further include:
Step 7: Strain of Beauveria bassiana is purified and is saved: the beauveria bassiana obtained through step 6 is used
PDA slant medium is maintained in the refrigerator that temperature is 4 DEG C.
The method as described above for trapping acquisition beauveria bassiana using cylas formicarius, wherein described in step 3
The cylas formicarius adult raised through step 1 is placed on the pedotheque through step 2 acquisition, specifically:
The cylas formicarius adult raised through step 1 is placed in the transparent modeling equipped with the pedotheque acquired through step 2
Expect to raise in octagonal prismatic bottle, the size of the transparent plastic illiciumverum prismatic bottle are as follows: bottom width × height=6.5cm × 13cm, soil
The thickness of sample is 5-10cm, and water content in soil sample is maintained at 50%, and pedotheque surface is placed with sweet potato 1-2, and confession is sweet
The small weevil imago feeding of potato makes the beauveria bassiana in pedotheque infect cylas formicarius.
The method as described above for trapping acquisition beauveria bassiana using cylas formicarius, it is preferable that described in step 1
The size of transparent plastic illiciumverum prismatic bottle are as follows: bottom width × height=6.5cm × 13cm, it is sweet in the transparent plastic illiciumverum prismatic bottle
The quantity of the small weevil of potato is 10-20 head.
The method provided by the invention for trapping acquisition beauveria bassiana using cylas formicarius, has the advantages that
1, acquisition beauveria bassiana is trapped using cylas formicarius adult, can separates within 5-7 days and obtain bacterial strain, reduces to go to
Man power and material's input cost of field investigation acquisition substantially increases the isolated efficiency of the acquisition of beauveria bassiana.
2, infection is trapped using cylas formicarius, has obtained fresh infected cylas formicarius of falling ill, overcome straight
It connects and is easy contaminated defect when being separated from the polypide of falling ill that field acquires.
3, acquisition beauveria bassiana is trapped using cylas formicarius, it is biggish by seasonal effect overcomes polypide acquisition of falling ill
Defect can carry out trapping separation after any season acquires soil sample.
Specific embodiment
Presently filed embodiment is described in detail below in conjunction with embodiment, whereby to the application how application technology hand
Section solves technical problem and reaches the realization process of technical effect to fully understand and implement.
Existing beauveria bassiana acquisition separation method has been unable to meet Strain of Beauveria bassiana resource and supper toxic strain
The needs of screening.Infection is trapped using cylas formicarius, passes through the infection conditions of different time, it may be determined that pathogenicity is strong
Bacterial strain.Therefore, targetedly infecting and being separately cultured is to obtain Strain of Beauveria bassiana, improve beauveria bassiana resource investigation
Isolated key is acquired with bacterial strain.The embodiment of the present invention provides a kind of side that acquisition beauveria bassiana is trapped using cylas formicarius
Method, comprising the following steps:
Step 1: cylas formicarius is raised: cylas formicarius being placed in transparent plastic illiciumverum prismatic bottle, is to raise with sweet potato
Material, is raised in 25 ± 1 DEG C of temperature environment indoors.
In the step, it is preferable that the size of the transparent plastic illiciumverum prismatic bottle of selection are as follows: bottom width × height=6.5cm ×
13cm, the quantity of the cylas formicarius in transparent plastic illiciumverum prismatic bottle are 10-20 head.
Step 2: collecting soil sample: selection soil sampling point acquires soil, soil in the plough horizon of sampled point
Acquisition acquires 0.5kg soil with a thickness of 5-7mm, each sampled point, the soil acquired in every 5 sampled points is mixed, as soil
Earth sample.
Specifically, (1) takes soil working tool: clean spade, small plastic bag, pen, label.(2) depth selection: the method for sampling
It is after rejecting surface layer stone and sundries, to cut pedotheque under tiltedly being gone out with spade, depth is arable layer on determining sampled point
Depth, general 20mm or so, sampling with a thickness of 5~7mm, the depth of each point, thickness are answered almost the same.Take 0.5kg left every time
The right side is packed into small plastic bag, writes label.5 soil samples are taken, are mixed.
Step 3: carrying out trapping infection processing to cylas formicarius: the cylas formicarius adult raised through step 1 is set
In on the pedotheque acquired through step 2, cultivated 5-7 days under the conditions of 25+1 DEG C, 16L:8D.
Step 4: production PDA culture medium: in every 1000mL water be added potato 200g, glucose 20g, agar 15~
20g is uniformly mixed.
Step 5: the separation of beauveria bassiana: the ball spore on the cylas formicarius polypide obtained through step 3 culture is white
Stiff bacterium mycelia and conidium separate from cylas formicarius polypide.
Step 6: the purifying culture of beauveria bassiana: by the beauveria bassiana mycelia obtained through step 5 and conidium
It is transferred to and in PDA culture medium made from step 4, is cultivated 3-4 days in 25+2 DEG C of temperature environment, then step 6 is repeated to grasp
Make 2-3 times, the beauveria bassiana purified.
The method as described above for trapping acquisition beauveria bassiana using cylas formicarius, it is preferable that after step 6,
Can also include:
Step 7: Strain of Beauveria bassiana is purified and is saved: the beauveria bassiana obtained through step 6 is used
PDA slant medium is maintained in the refrigerator that temperature is 4 DEG C.
The method as described above for trapping acquisition beauveria bassiana using cylas formicarius, wherein described in step 3
The cylas formicarius adult raised through step 1 is placed on the pedotheque through step 2 acquisition, is specifically as follows:
The cylas formicarius adult raised through step 1 is placed in the transparent modeling equipped with the pedotheque acquired through step 2
Expect to raise in octagonal prismatic bottle, the size of the transparent plastic illiciumverum prismatic bottle are as follows: bottom width × height=6.5cm × 13cm, soil
The thickness of sample is 5-10cm, and water content in soil sample is maintained at 50%, and pedotheque surface is placed with sweet potato 1-2, and confession is sweet
The small weevil imago feeding of potato makes the beauveria bassiana in pedotheque infect cylas formicarius.
Given below is the Application Example of technical solution provided by the invention: it is white that cylas formicarius traps acquisition ball spore
Stiff bacterium.
1. the raising and infection of cylas formicarius
Using transparent plastic illiciumverum prismatic bottle (bottom width × height=6.5cm × 13cm), the soil-like of acquisition is added in bottle
Product, soil horizon thickness 5-10cm place into sweet potato 1, then access cylas formicarius adult 10-20 head, are placed in 25-28 DEG C of condition
Lower raising.It is primary every observation for 24 hours after cylas formicarius adult is accessed the transparent plastic illiciumverum prismatic bottle containing soil sample,
The infected situation for observing and recording cylas formicarius adult is taken out after discovery has infected cylas formicarius individual
Be placed in bottom be covered with one sterilizing and on the filter paper of sterile water infiltration, trained under the conditions of 25 ± 1 DEG C, 16L:8D in illumination cultivation
It supports, to grow the mycelia and conidium of a large amount of beauveria bassiana.
2. the separation of beauveria bassiana
The beauveria bassiana mycelia and conidium that the cylas formicarius polypide surface of infection is taken with the oese libation at an ancient wedding ceremony, with scribing line
Method is seeded in PDA culture medium surface, and inoculation postposition is cultivated under the conditions of 25 ± 1 DEG C, 16L:8D in the light incubator, for ball spore
Muscardine growth, is inoculated with 5 wares every time.
3. interpretation of result
After to Yunnan different regions farmland and forest soil acquisition, after accessing cylas formicarius adult in the soil sample.
The result shows that the average recall rate of beauveria bassiana is up to 78.5%, and cylas formicarius is by beauveria bassiana in the soil sample of acquisition
Average rate crosses 80.5%.It traps result of infection and is shown in Table 1.
Table 1 traps infection conditions table
In conclusion the method provided in an embodiment of the present invention for trapping acquisition beauveria bassiana using cylas formicarius, leads to
It crosses and traps infection using cylas formicarius, overcome and be easy to be contaminated when directly being separated from the polypide of falling ill that field acquires
Defect, trap acquisition beauveria bassiana using cylas formicarius, overcome polypide of falling ill and acquire the defect big by seasonal effect,
Therefore technical solution provided by the invention is a kind of efficient, quick beauveria bassiana acquisition method.
Above description shows and describes several preferred embodiments of the present application, but as previously described, it should be understood that the application
Be not limited to forms disclosed herein, should not be regarded as an exclusion of other examples, and can be used for various other combinations,
Modification, and can be in the application contemplated scope, modifications can be made through the above teachings or related fields of technology or knowledge.And this
The modifications and changes that field personnel are carried out do not depart from spirit and scope, then all should be in the application appended claims
Protection scope in.
Claims (4)
1. a kind of method for trapping acquisition beauveria bassiana using cylas formicarius, which comprises the following steps:
Step 1: cylas formicarius is raised: cylas formicarius is placed in transparent plastic illiciumverum prismatic bottle, using sweet potato as feed,
It is raised in 25 ± 1 DEG C of temperature environment indoors；
Step 2: collecting soil sample: selection soil sampling point acquires soil, soil collection in the plough horizon of sampled point
With a thickness of 5-7mm, each sampled point acquires 0.5kg soil, the soil acquired in every 5 sampled points is mixed, as soil-like
Product；
Step 3: trap infection processing to cylas formicarius: by the cylas formicarius adult raised through step 1 be placed in through
On the pedotheque of step 2 acquisition, cultivated 5-7 days under the conditions of 25+1 DEG C, 16L:8D；
Step 4: production PDA culture medium: potato 200g, glucose 20g, 15~20g of agar are added in every 1000mL water,
It is uniformly mixed；
Step 5: the separation of beauveria bassiana: by the beauveria bassiana on the cylas formicarius polypide obtained through step 3 culture
Mycelia and conidium separate from cylas formicarius polypide；
Step 6: the purifying culture of beauveria bassiana: the beauveria bassiana mycelia obtained through step 5 and conidium are transferred to
It in PDA culture medium made from step 4, is cultivated 3-4 days in 25+2 DEG C of temperature environment, then by step 6 repetitive operation 2-3
Beauveria bassiana that is secondary, being purified.
2. the method for trapping acquisition beauveria bassiana using cylas formicarius as described in claim 1, which is characterized in that in step
After rapid six, further includes:
Step 7: Strain of Beauveria bassiana is purified and is saved: is oblique using PDA to the beauveria bassiana obtained through step 6
Face culture medium is maintained in the refrigerator that temperature is 4 DEG C.
3. the method for trapping acquisition beauveria bassiana using cylas formicarius as described in claim 1, which is characterized in that step
The cylas formicarius adult raised through step 1 is placed on the pedotheque through step 2 acquisition described in three, specifically:
The cylas formicarius adult raised through step 1 is placed in the transparent plastic eight equipped with the pedotheque acquired through step 2
It is raised in the prismatic bottle of angle, the size of the transparent plastic illiciumverum prismatic bottle are as follows: bottom width × height=6.5cm × 13cm, pedotheque
Thickness be 5-10cm, water content in soil sample is maintained at 50%, and pedotheque surface is placed with sweet potato 1-2, small for sweet potato
Weevil imago feeding makes the beauveria bassiana in pedotheque infect cylas formicarius.
4. the method as described in any one of claims 1-3 for trapping acquisition beauveria bassiana using cylas formicarius, feature
It is, the size of the prismatic bottle of transparent plastic illiciumverum described in step 1 are as follows: bottom width × height=6.5cm × 13cm, the transparent modeling
The quantity for expecting the cylas formicarius in octagonal prismatic bottle is 10-20 head.
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Cited By (2)
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| CN115074258A (en) * | 2022-07-18 | 2022-09-20 | 广东海洋大学 | Beauveria bassiana Bals-1722 and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114317289A (en) * | 2022-01-11 | 2022-04-12 | 云南农业大学 | Beauveria bassiana Bbsfa202007 strain and application thereof |
| CN114317289B (en) * | 2022-01-11 | 2022-09-16 | 云南农业大学 | Beauveria bassiana Bbsfa202007 strain and application thereof |
| CN115074258A (en) * | 2022-07-18 | 2022-09-20 | 广东海洋大学 | Beauveria bassiana Bals-1722 and application thereof |
| CN115074258B (en) * | 2022-07-18 | 2024-03-15 | 广东海洋大学 | Beauveria bassiana Bals-1722 and application thereof |
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