CN115747130A - Culture medium for promoting spore production of metarhizium limacinum Mr006, preparation and application thereof - Google Patents
Culture medium for promoting spore production of metarhizium limacinum Mr006, preparation and application thereof Download PDFInfo
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Abstract
The invention discloses a culture medium for promoting spore production of metarhizium lepigone Mr006, and a preparation and application thereof, and belongs to the technical field of biology. The culture medium comprises the following components in parts by mass: 20-60 parts; rice hull: 10-20 parts; malt: 2-4 parts; yellow powder: 8-16 parts; yeast powder: 2-8 parts; water: 30-60 parts; freeze-drying worm powder: 1-2 parts; zinc phosphate: 0.1 to 0.2 portion; ferrous sulfate: 0.05 to 0.15 portion; the lyophilized powder of the insect body is lyophilized powder of Spodoptera frugiperda. Metarhizium limerimetarhizium rileyi The preservation number of Mr006 is CGMCC NO.40171, and the Mr006 is preserved in China general microbiological culture Collection center at 5 months and 9 days 2022 in China Committee for culture Collection of microorganismsThe hidden address is as follows: xilu No. 1 Hospital No. 3, beijing, chaoyang, beicheng. Component screening of a culture medium and optimization design of culture conditions are performed on a strain Mr006 separated from stiff armyworm in a corn field, so that a culture method for better promoting spore production of the Metarrhizium Laplace Mr006 is found, and support is provided for application of the Metarrhizium Laplace Mr 006.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a culture medium for promoting spore production of Metarrhizium Launderii Mr006, a preparation and an application thereof, in particular to a culture method for promoting spore production of Metarrhizium Launderii, which is applied to prevention and control of Lepidoptera pests such as Spodoptera frugiperda and the like.
Background
At present, the harm of pests to crops is one of the major factors causing agricultural yield reduction. The long-term use of a large amount of chemical pesticides can kill pests and bring a series of side effects of pesticide residue exceeding standards of agricultural products, year-by-year increase of pest resistance level, environmental pollution and the like. In view of this, biological control is being increasingly appreciated, and pest control using entomopathogenic microorganisms is one of the important means for biological control of pests. Compared with other insecticidal microorganisms, the insecticidal fungi have the advantages of easy popularization, environmental friendliness, difficulty in generating drug resistance and the like, and play an important role in biological control of pests.
Metarhizium rileyi (Metarhizium rileyi) is an entomopathogenic fungus widely distributed in China, can infect various lepidoptera pests, and particularly has extremely high infectivity on noctuidae pests such as spodoptera frugiperda, spodoptera exigua, soybean looper, prodenia litura and the like, thereby having higher development and utilization values.
The spore yield, the spore production time and the like are important factors influencing the application of the metarhizium lepigone strain. Currently, agricultural and sideline products such as wheat bran, corn flour, rice flour and the like which are cheap are mostly adopted in the green muscardine fungus fermentation substrate to obtain the maximum spore yield. In optimizing the substrate combination, the shortening of the sporulation time is hardly considered as an index, and only the spore yield is targeted.
The most commonly used culture media for culturing the metarhizium reinhardtii in a laboratory are a sass maltose-yeast culture medium and a potato culture medium, but the problems of low sporulation rate, low spore germination rate, long sporulation period, high cost and the like still exist. At present, few research reports about the spore yield and spore production time of the metarhizium reinhardtii by inorganic salt metal ions and polypide tissues are reported.
The metarhizium lepigone Mr006 is obtained by separating and identifying metarhizium lepigone infected with entomogenous fungi in fields discovered by the biological control team of the institute of plant protection and agricultural product quality safety of agricultural academy of agricultural sciences of Anhui province in Taihe corn growing area. The specific sequencing results are as follows (540 bp):
experiments show that the Spodoptera frugiperda, the ostrinia nubilalis and the like obtained by separation in the laboratory have better effects. Based on the above, through adding inorganic salt metal ions, polypide tissues and the like, a solid fermentation spore production test of different components of the culture medium of the metarhizium rebaudigenum Mr006 is performed, so that the production process is simplified, the spore production time is shortened, the spore yield is improved, the production cost is reduced, and the culture method for promoting the spore production of the metarhizium rebaudigenum Mr006 is obtained.
Disclosure of Invention
1. Problems to be solved
In view of the problems of the prior art, the object is to provide a culture method for promoting the spore production of Metarhizium reinhardtii Mr006 so as to provide support for late fermentation production.
The above purpose is realized by the following scheme:
a culture method for promoting the spore production of Metarrhizium Lateorum Mr006 is characterized in that: the method comprises the following steps:
(1) Weighing the following components in parts by weight and mixing uniformly: 40 parts of rice, 15 parts of rice hull, 3 parts of maltose, 12 parts of yellow powder, 5 parts of yeast powder, 25 parts of water, 1.5 parts of spodoptera frugiperda freeze-dried powder, 0.15 part of zinc phosphate, 0.1 part of ferrous sulfate and 23.5 parts of water;
(2) Standing and soaking the uniformly mixed raw materials for 4 hours, bagging the raw materials, sterilizing the raw materials for 50 minutes at 121 ℃, inoculating the green muscardine fungus Mr006 liquid seed bacterial liquid of the green muscardine fungus to the raw materials after the temperature is reduced to 30 ℃, stirring the raw materials and the seed bacterial liquid of the green muscardine fungus Mr006 liquid seed bacterial liquid of the green muscardine fungus with the inoculum size of 10 percent, and uniformly mixing the raw materials
Culturing in a light incubator at 28 deg.C (16L/8D, light and dark alternation), and culturing for 3D; adjusting temperature to 24 deg.C, irradiating with light (23L/1D, light and dark alternately), culturing for 14 days, stopping fermentation, and sampling at regular time.
2. Technical scheme
In order to solve the above problems, the present invention adopts the following technical solutions.
A culture medium for promoting the spore production of Metarrhizium Lateorum Mr006 comprises the following components in parts by mass,
the insect body freeze-dried powder is Spodoptera frugiperda insect body freeze-dried powder.
The culture medium comprises the following components in parts by mass,
the insect body freeze-dried powder is Spodoptera frugiperda insect body freeze-dried powder.
The culture medium comprises the following components in parts by mass,
the insect body freeze-dried powder is Spodoptera frugiperda insect body freeze-dried powder.
Any one of the culture media, the metarhizium rileyi Mr006 of the metarhizium laevis has a preservation number of CGMCC NO.40171, and is preserved in China general microbiological culture Collection center at 5-9 months in 2022 at the preservation address of: xilu No. 1 Hospital No. 3, beijing, chaoyang, north.
A preparation of Metarrhizium Laevigatum Mr006 is prepared by culturing in any culture medium and adding adjuvant, and is in the form of suspension.
Use of a medium according to any one of the preceding claims for promoting sporulation.
The application of the metarhizium rebaudianum Mr006 preparation in promoting sporulation.
3. Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
metarhizium reilianum is an entomopathogenic fungus with strong pathogenicity to larvae of the noctuidae family, and often causes high-intensity insect epidemic diseases in nature. Most of the research works of fungi require indoor artificial culture of the fungi, and different fungi have large requirements on nutrient compositions of culture media. The culture of the Metarrhizium limacinum generally adopts a Saco ura malt agar medium (SMAY), and the culture medium is balanced in terms of mycelial growth, sporulation yield, sporulation speed and the like of the fungus, but has a long growth cycle and low sporulation yield. Different strains may also respond differently to culture components, and thus the optimal medium composition may vary from strain to strain. In view of this, in order to better develop the related research on the metarhizium reilianum, component screening and culture condition optimization design of a culture medium are performed on a strain Mr006 separated from the stiff worm of spodoptera frugiperda in the corn field, so as to find a culture method for better promoting spore production of the metarhizium reilianum Mr006, and support is provided for application of the metarhizium reilianum Mr 006.
In order to obtain a culture method for better promoting the spore production of the metarhizium reinhardtii Mr006, on the basis of optimizing a laboratory culture medium and culture conditions, screening pilot-scale fermentation components, amplifying culture and analyzing the influence on the spore production starting time and the spore production quantity.
The invention provides a culture method for promoting the spore production of Metarrhizium Lateucrium Mr006, which comprises the following steps:
(1) Weighing the following raw materials in parts by weight (kg) and mixing uniformly: 40 parts of rice, 15 parts of rice hull, 3 parts of maltose, 12 parts of yellow powder, 5 parts of yeast powder, 25 parts of water, 1.5 parts of spodoptera frugiperda freeze-dried powder, 0.15 part of zinc phosphate, 0.1 part of ferrous sulfate and 23.5 parts of water;
(2) Standing and soaking the uniformly mixed raw materials for 4 hours, bagging the raw materials, sterilizing the raw materials for 50 minutes at 121 ℃, inoculating the green muscardine fungus Mr006 liquid seed bacterial liquid of the green muscardine fungus to the raw materials after the temperature is reduced to 30 ℃, stirring the raw materials and the seed bacterial liquid of the green muscardine fungus Mr006 liquid seed bacterial liquid of the green muscardine fungus with the inoculum size of 10 percent, and uniformly mixing the raw materials
Culturing in a 28 ℃ illumination incubator (16L/8D, alternate light and dark), and culturing for 3 days; adjusting temperature to 24 deg.C, irradiating with light (23L/1D, light and dark alternately), culturing for 14 days, stopping fermentation, and sampling at regular time.
(3) Washing off conidia collected on a solid fermentation culture medium by using a Tween-80 solution to obtain a conidia solution, preparing a diluent with a proper concentration by using a 10-time dilution method, and observing and calculating the spore amount under a microscope by using a cell counting method.
Detailed Description
The invention is further described with reference to specific examples.
1. Materials and methods
Material
Metarhizium leydii Mr006 was isolated from Spodoptera frugiperda and stored in the laboratory.
Sass maltose-yeast medium: maltose 4%, peptone 1%, yeast extract powder 1%, agar 1.5%.
Spodoptera frugiperda polypide freeze-dried powder: is prepared from 6-instar larva through ultralow-temp freeze drying, adding liquid nitrogen, grinding and pulverizing.
Liquid seed culture medium: maltose 4%, peptone 1%, yeast extract powder 1%, zinc sulfate 0.15%, and ferrous sulfate 0.1%.
Raw materials for solid fermentation medium and inorganic salts: rice, rice hull, maltose, soybean meal, yeast powder, water, zinc sulfate, ferrous sulfate and the like.
The main equipment is as follows: a steam sterilizer, an ultra-clean workbench, a temperature-controlled shaking table, a temperature-controlled biochemical incubator, a balance, a microscope, a spore collector and the like.
Method
Start time of sporulation: and (3) during the growth of hyphae, immediately picking out green powder, inspecting under a microscope, and determining as conidium, namely determining as the spore production starting time.
And (3) counting spores: scraping the spore powder collected after fermentation from the culture medium (material) or collecting with spore collector, placing into 100mL conical flask, adding sterile water containing 0.05% Tween-80, stirring for 30min on magnetic stirrer to fully disperse the spores, preparing into uniform spore suspension, and determining spore amount by blood cell plate counting method.
LC 50 Value determination: the collected spore powder was prepared into 1X 10 with sterile water containing 0.05% Tween-80 4 1X 10 units/mL 5 1X 10 units/mL 6 1X 10 units/mL 7 1X 10 units/mL 8 Spore suspensions at five concentrations per mL. 20 heads of 3-instar spodoptera frugiperda larvae were treated at each concentration, with 0.05% tween-80 water as a control. Inoculating bacterial liquids with different concentrations, placing in an artificial climate insect breeding room for breeding, and observing and recording the number of dead insects for 10 days. Repeat 3 times. Calculating LC 50 The value is obtained.
The result of the infection rate of the metarhizium lepigone Mr006 spores is as follows: respectively culturing with optimal combination and common combination to obtain Metarrhizium Laevigatum Mr006 spore powder with concentration of 1 × 10 4 1X 10 units/mL 5 1X 10 units/mL 6 1X 10 units/mL 7 1X 10 units/mL 8 Each mL of five spore suspensions at concentrations, for a total of 10 treatments, 20 beetles per treatment, 4 replicates. Selecting 3-instar spodoptera frugiperda larvae with consistent growth, treating the spore suspension by an insect soaking method, and detecting the germ carrying rate of the larvae at 24 hours.
The result of the experiment of the pot culture of the Metarrhizium Laevigatum Mr 006: respectively culturing the optimum combination and the common combination to obtain the Metarrhizium Laevigatum Mr006 spore powder according to the mass percentage
The preparation method comprises the following steps: according to the formula, grinding solid additives such as an effective component thickener and the like in an oil phase carrier through a sand mill until the particle size reaches 2-3 microns, adding the rest additive substances according to the formula, then adding the metarhizium reinhardtii Mr006 spore powder, adjusting to the required concentration, fully shearing, and uniformly stirring to obtain the finished product.
200 corn seedlings obtained by indoor pot culture are inoculated into 3-instar spodoptera frugiperda larvae, and 2 heads of each corn seedling are planted. The dispersible oil suspension agent of 10 hundred million spores/ml metarhizium reinhardtii Mr006 is sprayed on stems and leaves according to the dosage of 100 g of the preparation for each mu, and the control effect is investigated in 5, 7, 14 and 21 days respectively.
2. Results and analysis
Optimization of culture conditions for promoting spore production of Metarrhizium Laelii Mr006 spore production
On the basis of the earlier stage test, a Sasa maltose-yeast culture medium (SMAY) is adopted and respectively set to (1) a 28 ℃ illumination incubator for culture (16L/8D, light and dark alternation) and culture for 14D; (2) Culturing in a 28 ℃ illumination incubator (16L/8D, alternate light and dark), and culturing for 3 days; adjusting the temperature to 24 ℃, illuminating (23L/1D, alternating light and dark), continuously culturing for 14D; and comparing the spore yields.
The metarhizium lepigo Mr006 grows well under the culture condition (2), the spore production starting time is early, and the spore production reaches 8.2x10 after the metarhizium lepigo is cultured for 14 days 8 Per cm 2 And the spore yield under the culture condition (1) is 1X 10 8 Per cm 2 (ii) a . Therefore, the temperature is reduced in the later period, the spore production is facilitated under the dark condition, the culture condition (2) is obviously superior to the culture condition (1), and the subsequent experiment is determined to be carried out under the culture condition (2).
Optimization of culture components for promoting spore production of Metarrhizium Laevigatum Mr006
On the basis of the early test, respectively adding a proper amount of inorganic salt and insect freeze-dried powder (see table 1) into a Sasa maltose-yeast culture medium (SMAY), taking the non-added Sasa maltose-yeast culture medium (SMAY) as a reference, culturing in a light incubator (16L/8D, alternating light and dark) at 28 ℃ according to the culture condition (2), and culturing for 3 days; adjusting the temperature to 24 ℃, illuminating (23L/1D, alternating light and dark), continuously culturing for 14D; during the period, the time for starting sporulation is determined, and the final sporulation amount is compared.
TABLE 1 optimization of the composition of the culture Medium
As can be seen from the results in Table 1, the addition of various inorganic salts and lyophilized powder of insect bodies to the Sasa maltose-yeast medium (SMAY) resulted in the onset of sporulation, sporulation and LC production of Metarrhizium Lateorum Mr006 50 The values all have an influence. Calcium chloride inhibits growth, the start time of sporulation is delayed, the sporulation quantity is obviously reduced, and LC 50 The value rises significantly; dipotassium phosphate slightly influences spore yield; manganese sulfate can be produced in advanceStart time of spore, vs. LC 50 The value influence is not great; the zinc sulfate advances the spore production starting time from 8 days to 4 days, and the spore production is improved along with the increase of the addition amount, and the maximum spore production can reach 8.6 multiplied by 10 8 Per cm 2 ，LC 50 The value slightly decreased; the ferrous sulfate advances the spore production starting time from 8 days to 5 days, and the spore production is obviously improved to 8.8 multiplied by 10 at most 8 Per cm 2 ，LC 50 The value is slightly reduced; the addition of the worm freeze-dried powder can also shorten the start time of spore production and improve the spore yield which can reach 9.3 multiplied by 10 to the maximum 8 Per cm 2 Can obviously reduce LC 50 The value is obtained.
Therefore, zinc sulfate, ferrous sulfate and Spodoptera frugiperda worm freeze-dried powder can obviously shorten the spore production starting time, improve the spore production quantity and improve the toxicity of the Mr006 strain. .
Metarhizium reilianum Mr006 solid fermentation process
According to the component ratio shown in the table 2, the culture medium containing the Spodoptera frugiperda worm freeze-dried powder is optimized, and the optimal solid fermentation process suitable for the Metarrhizium Latifolii Mr006 is determined by taking the spore production starting time and the spore production amount as indexes.
Inoculating the preserved Metarrhizium Laevigatum Mr006 into liquid seed culture medium, culturing for 3-5 days, and stopping culturing when the culture solution is turbid and mycelial pellets appear. Uniformly mixing the raw materials treated according to the table 2, standing and soaking for 4 hours, bagging the mixture for 50 minutes at 121 ℃, carrying out autoclaving at the temperature of the mixture, inoculating metarhizium lepigonense Mr006 liquid seed bacteria liquid at the temperature of 30 ℃, carrying out 10 percent inoculation amount, stirring and uniformly mixing the mixture, carrying out fermentation culture at the temperature of 28 ℃ for 72 hours, then, reducing the temperature to 24 ℃, continuing the culture, determining the start time of spore production, stopping the fermentation until the spore production is complete, and sampling and checking the period at regular time. And (4) after fermentation is finished, collecting spore powder by using a spore collector, and calculating the spore yield.
TABLE 2 Metarrhizium laevigatum Mr006 solid fermentation test results
From the test results in table 2, it can be seen that: the solid fermentation culture mediums with different component ratios have the advantages of shortened spore production starting time of the Metarrhizium Laevigatum Mr006 and large influence difference of spore production, and the maximum spore treatment amount of 15 spores is 11.2 multiplied by 10 9 The minimum of the particles/g is 1.6 multiplied by 10 9 Per gram; the start time of sporulation was 4 days at the earliest and 9 days at the latest. LC (liquid Crystal) 50 The lowest value is 3.3X 10 6 one/mL, up to 8.94X 10 6 One per mL. Wherein the treatment 5 combination is optimal, the spore production time is shortened from 9 days to 4 days, and the spore yield can be improved to 11.2 multiplied by 10 9 Per gram. LC (liquid Crystal) 50 The value is 3.3X 10 6 one/mL, treatment 11 combination, spore production time is shortened from 9 days to 5 days, and spore yield can be increased to 10.2 × 10 9 Per gram. LC (liquid Crystal) 50 The value was 3.5X 10 6 one/mL.
Therefore, after the optimized component proportion such as zinc sulfate, ferrous sulfate, spodoptera frugiperda worm freeze-dried powder and the like is added, the beginning time of sporulation of the metarhizium lepigone Mr006 is shortened, the sporulation quantity is improved, and the toxicity to the spodoptera frugiperda is improved.
Results of infection rate of Metarrhizium Laevigatum Mr006 spores
From the results in table 3, it can be seen that: with the increase of the spore concentration of the metarhizium lepigone Mr006, the infection rate is correspondingly increased. The spore concentration of Metarrhizium Laevigatum Mr006 is higher than 1 × 10 5 When the spore/ml is increased, the spore infection rate obtained by optimal combined culture (adding zinc sulfate, ferrous sulfate and Spodoptera frugiperda polypide freeze-dried powder) is higher than that of the common combination (without the additives). Therefore, the zinc sulfate, the ferrous sulfate and the spodoptera frugiperda larva freeze-dried powder are added, so that the infection rate of the metarhizium lebonensis Mr006 spores to spodoptera frugiperda larvae is favorably improved.
TABLE 3 relationship between the invasion rate of Metarrhizium Laevigatum Mr006 to Spodoptera frugiperda larvae and spore concentration level
Test result of pot culture of Metarrhizium Laevigatum Mr006 spores
From the results in table 4, it can be seen that: 10 hundred million spores/ml metarhizium reinhardtii Mr006 dispersible oil suspending agent has better control effect on spodoptera frugiperda in a pot experiment. At the same time after the application, the control effect of the dispersible oil suspending agent of 10 hundred million spores/ml green muscardine fungus Mr006 processed by spores obtained by optimal combined culture (adding zinc sulfate, ferrous sulfate and spodoptera frugiperda polypide freeze-dried powder) is higher than that of the common composition (without the additives). Therefore, the zinc sulfate, the ferrous sulfate and the spodoptera frugiperda larva freeze-dried powder are added, so that the control effect of the metarhizium lepigone Mr006 spores on spodoptera frugiperda larvae is improved.
TABLE 4 application effect of Metarrhizium Laevigatum Mr006 spore in Spodoptera frugiperda prevention and treatment
And (4) conclusion: 40 parts of rice, 15 parts of rice hull, 3 parts of maltose, 12 parts of yellow powder, 5 parts of yeast powder, 25 parts of water, 1.5 parts of spodoptera frugiperda freeze-dried powder, 0.15 part of zinc phosphate, 0.1 part of ferrous sulfate and 23.5 parts of water; the composite is a better combination of the Metarrhizium Laevigatum Mr006 solid fermentation substrate; standing and soaking the uniformly mixed raw materials for 4 hours, bagging the raw materials, sterilizing the raw materials for 50 minutes at 121 ℃, inoculating metarhizium lepigone Mr006 liquid seed bacterial liquid after the temperature is reduced to 30 ℃, stirring the raw materials and the seed bacterial liquid uniformly, and then culturing the raw materials in a light incubator at 28 ℃ (16L/8D, alternating light and dark) for 3 days; adjusting temperature to 24 deg.C, irradiating with light (23L/1D, light and dark alternation), culturing, and stopping fermentation after culturing for 14D.
After the optimized component proportion such as zinc sulfate, ferrous sulfate, spodoptera frugiperda worm freeze-dried powder and the like is added, the beginning time of sporulation of the metarhizium laevigatum Mr006 is shortened, the sporulation quantity is improved, and the toxicity to the spodoptera frugiperda is improved. In the current literature reports, no relevant report is found.
Claims (7)
1. A culture medium for promoting spore production of Metarrhizium Laelii Mr006 is characterized in that:
the culture medium comprises the following components in parts by mass,
20-60 parts of rice;
10-20 parts of rice hulls;
2-4 parts of maltose;
8-16 parts of yellow powder;
2-8 parts of yeast powder;
30-60 parts of water;
1-2 parts of worm freeze-dried powder;
0.1 to 0.2 portion of zinc phosphate;
0.05 to 0.15 portion of ferrous sulfate;
the insect body freeze-dried powder is Spodoptera frugiperda insect body freeze-dried powder.
2. The culture medium of claim 1, wherein:
the culture medium comprises the following components in parts by mass,
30-50 parts of rice;
12-18 parts of rice hulls;
2-4 parts of maltose;
9-14 parts of yellow powder;
2-8 parts of yeast powder;
35-55 parts of water;
1-2 parts of worm freeze-dried powder;
0.1 to 0.2 portion of zinc phosphate;
0.05 to 0.15 portion of ferrous sulfate;
the insect body freeze-dried powder is Spodoptera frugiperda insect body freeze-dried powder.
3. The culture medium according to claim 1, wherein:
the culture medium comprises the following components in parts by mass,
40 parts of rice;
15 parts of rice hulls;
3 parts of maltose;
12 parts of yellow powder;
5 parts of yeast powder;
48.5 parts of water;
1.5 parts of worm freeze-dried powder;
0.15 part of zinc phosphate;
0.1 part of ferrous sulfate;
the insect body freeze-dried powder is Spodoptera frugiperda insect body freeze-dried powder.
4. The culture medium according to any one of claims 1 to 3, wherein: the Metarhizium reinhardtiimetarhizium rileyi The preservation number of Mr006 is CGMCC NO.40171, and the Mr006 is preserved in the China general microbiological culture Collection center at 5-9 months in 2022 at the preservation address: xilu No. 1 Hospital No. 3, beijing, chaoyang, north.
5. A metarhizium reinhardtii Mr006 preparation is characterized in that: prepared by culturing the culture medium of any one of claims 1 to 4 with the aid of an auxiliary agent, and the preparation is in the form of a suspension.
6. Use of a medium according to any of claims 1 to 4 for promoting sporulation.
7. Use of the preparation of Metarhizium lebbeck Mr006 according to claim 5 for promoting sporulation.
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| CN116555050A (en) * | 2023-06-05 | 2023-08-08 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Method for propagating Metarhizium anisopliae Mr006 and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050095259A1 (en) * | 2003-11-04 | 2005-05-05 | Liu Chien Y. | Manufacturing method for entomopathogenic fungi |
| WO2009035925A2 (en) * | 2007-09-13 | 2009-03-19 | The United States Of America, As Represented By The Secretary Of Agriculture | Composition of entomopathogenic fungus and method of production and application for insect control |
| CN103087924A (en) * | 2012-11-27 | 2013-05-08 | 河南省农业科学院 | Entomogenous fungus Nomuraearileyi, as well as culture method and application thereof |
| CN104673740A (en) * | 2015-03-10 | 2015-06-03 | 聊城大学 | Culture medium for promoting spore production of nomuraea rileyi and preparation method |
| CN111235039A (en) * | 2020-03-04 | 2020-06-05 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Culture medium for culturing metarhizium anisopliae with high toxicity, preparation and application thereof |
| CN111471600A (en) * | 2020-05-08 | 2020-07-31 | 北京化工大学 | Culture medium for preparing metarhizium anisopliae spore powder and preparation method of metarhizium anisopliae high spore powder |
| CN111662828A (en) * | 2020-06-30 | 2020-09-15 | 福建省农业科学院植物保护研究所 | Metarhizium leylanicum and application thereof |
-
2022
- 2022-05-25 CN CN202210572361.8A patent/CN115747130B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050095259A1 (en) * | 2003-11-04 | 2005-05-05 | Liu Chien Y. | Manufacturing method for entomopathogenic fungi |
| WO2009035925A2 (en) * | 2007-09-13 | 2009-03-19 | The United States Of America, As Represented By The Secretary Of Agriculture | Composition of entomopathogenic fungus and method of production and application for insect control |
| CN103087924A (en) * | 2012-11-27 | 2013-05-08 | 河南省农业科学院 | Entomogenous fungus Nomuraearileyi, as well as culture method and application thereof |
| CN104673740A (en) * | 2015-03-10 | 2015-06-03 | 聊城大学 | Culture medium for promoting spore production of nomuraea rileyi and preparation method |
| CN111235039A (en) * | 2020-03-04 | 2020-06-05 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Culture medium for culturing metarhizium anisopliae with high toxicity, preparation and application thereof |
| CN111471600A (en) * | 2020-05-08 | 2020-07-31 | 北京化工大学 | Culture medium for preparing metarhizium anisopliae spore powder and preparation method of metarhizium anisopliae high spore powder |
| CN111662828A (en) * | 2020-06-30 | 2020-09-15 | 福建省农业科学院植物保护研究所 | Metarhizium leylanicum and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| PRAKASH G V S B 等: "Statistical optimization of process variables for the large-scale production of Metarhizium anisopliae conidiospores in solid-state fermentation", 《BIORESOURCE TECHNOLOGY》, vol. 99, no. 6, pages 1530 - 1537, XP022410772, DOI: 10.1016/j.biortech.2007.04.031 * |
| SONG Z 等: "Statistical optimisation of process variables and large-scale production of Metarhizium rileyi (Ascomycetes: Hypocreales) microsclerotia in submerged fermentation", 《MYCOLOGY》, vol. 8, no. 1, pages 9 - 47 * |
| 刘磊 等: "绿僵菌在3种不同培养基上培养时产孢量的影响因素", 《热带作物学报》, vol. 31, no. 3, pages 469 - 473 * |
| 杨淑仪 等: "莱氏野村菌产孢培养基的正交优化设计", 《湖北农业科学》, vol. 55, no. 23, pages 6164 - 6168 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116555050A (en) * | 2023-06-05 | 2023-08-08 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Method for propagating Metarhizium anisopliae Mr006 and application thereof |
| CN116555050B (en) * | 2023-06-05 | 2024-01-05 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Method for propagating Metarhizium anisopliae Mr006 and application thereof |
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