CN108753630B - Paecilomyces gossypii strain Sanju-18 and application thereof - Google Patents
Paecilomyces gossypii strain Sanju-18 and application thereof Download PDFInfo
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- CN108753630B CN108753630B CN201810849174.3A CN201810849174A CN108753630B CN 108753630 B CN108753630 B CN 108753630B CN 201810849174 A CN201810849174 A CN 201810849174A CN 108753630 B CN108753630 B CN 108753630B
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- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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Abstract
The invention relates to the field of biological pharmacy, in particular to a preparation method and application of a microbial agent. The invention provides a Paecilomyces gossypii (Paecilomyces heliothis) strain Sanju-18, which is preserved in 14 days 06 and 14 months 2018 by China general microbiological culture Collection center (CGMCC for short), wherein the preservation number is CGMCC No.15869, and the preservation address is No. 3 of Xilu No.1 of the morning district of the morning of Beijing City. The strain has good effect on preventing and treating potato scab, and can be used for biological prevention and treatment of potato scab.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to the technical field of paecilomyces gossypii (Paecilomyces hepialitis).
Background
The potato scab is a plant disease damaging potato tubers. The appearance of the disease is that the surface of the tuber is nearly round to irregular, suberized scab-like light brown scab or plaque, and the hand feel is rough. Generally, two types of disease symptoms are developed, including a reticulated lesion and a lacerated lesion. Generally, although the disease spots are only limited to the skin layer, the quality and the yield of the damaged potato blocks can be reduced, the damaged potato blocks are not storage-resistant, the appearance of the damaged potato blocks is not elegant, the commodity grade is greatly reduced, and certain economic loss is caused. In China, the incidence of diseases is low, the area of damage is small, and therefore people do not pay attention to the disease. However, in recent years, the disease incidence area in inner Mongolia, Hebei, Shandong and other areas is increasing, and the economic loss caused by the disease incidence area is also considerable. Therefore, scientific control of such diseases is urgently required.
According to the long-term separation and identification research on pathogenic microorganisms, pathogenic bacteria causing potato scab are mainly streptomyces scabicus and the like. And the pathogenic bacteria belong to conditional pathogenic bacteria, namely the pathogenic bacteria easily grow and cause potato diseases under the alkaline condition of the soil environment, and rarely appear in the acidic soil environment.
The control method mainly comprises the following steps: (1) selecting seed potatoes without diseases; (2) applying organic fertilizer or green manure more times can partially inhibit diseases; (3) performing rotation with Cucurbitaceae, Leguminosae, and Liliaceae vegetables for more than 5 years; (4) selecting a vegetable field with good water retention for planting, and watering in time when meeting drought in a potato bearing period; (5) and chemical prevention: some chemical pesticides can be used for preventing and treating scab; (6) biological prevention and treatment method: the method screens microorganisms with good inhibition effect on scab pathogenic bacteria and strong planting capability, and is applied in the potato seed production process and the farmland planting process to achieve the effect of inhibiting the growth of the pathogenic bacteria.
Disclosure of Invention
In order to effectively prevent and treat potato scab, the first purpose of the invention is to provide a Paecilomyces gossypii (Paecilomyces heliothis) strain Sanju-18, which is preserved in 14 days 06 and 14 months 2018 to China general microbiological culture Collection center (CGMCC for short), with the preservation number of CGMCC No.15869 and the preservation address of No. 3 Hospital No.1 Xilu north Chen in the sunward area of Beijing.
The second purpose of the invention is to provide a microbial agent, the active ingredient of which is the paecilomyces gossypii strain Sanju-18.
The third purpose of the invention is to provide a method for preparing microbial agent, 1) preparing potato solid culture medium (g/L): 1L of potato extract and 20g of glucose; adding 15g of agar into the solid culture medium; 2) paecilomyces gossypii (Paecilomyces heliothis) strain Sanju-18 was activated 3 times on potato solid medium; 3) culturing 1 ring in 50ml potato liquid culture medium at 28 deg.C for 2 days; 4) after the fermentation liquor is centrifuged for 1min at 12,000r/min, the supernatant and the precipitate are respectively collected and suspended in sterile water with the same volume to prepare cell suspension.
The fourth purpose of the invention is to provide the application of the bacterial strain Sanju-18 and the bacterial agent thereof, which can inhibit the growth of Streptomyces scabies (Streptomyces scabies), Streptomyces tumefaciens (S.Turginicabis) and Streptomyces galileoparus (S.galilaeus) and can effectively prevent and treat potato scab.
The microbial agent has the following advantages: 1) the microbial agent is a pure biological agent, is harmless to human bodies, does not pollute the environment, and can solve the environmental problem caused by chemical pesticide pollution; meanwhile, after the microbial agent is applied to soil, the soil quality can be improved, and the microbial agent has an active effect on reducing the occurrence of diseases. 2) The microbial agent can effectively prevent and treat potato scab. .
Drawings
FIG. 1 is a graph showing the bacteriostatic effect of the strain Sanju-18 on Streptomyces scabies 4.0076.
FIG. 2 is a graph showing the bacteriostatic effect of the strain Sanju-18 on Streptomyces scabies 4.1765.
FIG. 3 is a graph showing the bacteriostatic effect of the strain Sanju-18 on Streptomyces scabies 4.1789.
FIG. 4 is a graph showing the bacteriostatic effect of the strain Sanju-18 on Streptomyces albidoflauvs 4.3598.
FIG. 5 is a graph showing the bacteriostatic effect of the strain Sanju-18 on Streptomyces scabies.
FIG. 6 is a diagram showing the bacteriostatic effect of the three strains of torch-18 and 5 mixed strains of Streptomyces scabies 4.0076, Streptomyces scabies 4.1765, Streptomyces scabies acidovorans 4.1789, Streptomyces albidoflavus 4.3598 and Streptomyces scabies Europe during co-culture.
Detailed Description
Example 1
The preparation method of the microbial agent comprises the following steps:
1) potato liquid Medium (g/L) preparation: 1L of potato extract and 20g of glucose;
adding 15g of agar into the liquid potato culture medium to obtain a solid potato culture medium;
2) paecilomyces gossypii (Paecilomyces heliothis) strain Sanju-18 was activated 3 times on potato solid medium;
3) culturing 1 ring in 50ml potato liquid culture medium at 28 deg.C for 2 days;
4) after the fermentation broth was centrifuged at 12,000r/min for 1min, the supernatant was discarded, and the pellet was suspended in the same volume of sterile water to prepare a cell suspension.
Example 2
Preparation of a culture medium:
1) potato solid Medium (g/L): 1L of potato extract and 20g of glucose, wherein the pH value is natural; 15g of agar was added to the solid medium.
2) ISP-2 Medium (g/L): yeast extract 4g, malt extract 10g, glucose 4g, pH7.3, added 15g agar.
Example 3 strain Paecilomyces gossypii (Paecilomyces heliothis) triple torch-18 culture method:
the strain paecilomyces gossypii strain Sanju-18 is activated 3 times on a potato solid culture medium. Then, 1 ring of the suspension was placed in 50ml of potato broth and cultured on a shaker at 28 ℃ for 2 days. After the fermentation broth was centrifuged at 12,000r/min for 1min, the supernatant was discarded, and the pellet was suspended in the same volume of sterile water to prepare a cell suspension.
Example 4
The method for culturing pathogenic bacteria comprises the following steps:
1) streptomyces scabies (Streptomyces scabies) 4.0076: culturing on potato culture medium for 4-5 days at 28 deg.C.
2) Streptomyces scabies (Streptomyces acidiscacabes) 4.1789 and Streptomyces albidoflavus (Streptomyces albicorflavus) 4.3598 were cultured on ISP-2 medium for 4-5 days at 28 ℃.
Example 5
Preparation of pathogen suspension:
scraping spores and mycelia with 3ml of sterile water to prepare pathogen suspension with a certain concentration; a50 ul volume was required to coat the plates and allow the hyphae to grow and then spread over the surface of the plates.
Example 6
Single bacteriostasis test: streptomyces scabies (Streptomyces scabies)4.0076, Streptomyces scabies (Streptomyces scabies)4.1765, Streptomyces acidocaldarius (Streptomyces acidiscaibes) 4.1789, Streptomyces albidoflavus (Streptomyces albidoflavus)4.3598, and Streptomyces european scabies (Streptomyces scabies) each 50ul were spread on different plates (PDA, ISP-2), and 10ul of Paecilomyces gossypii (Paecilomyces helionsis) was spotted locally on the plates. Culturing at 28 ℃. Observed after 2 days.
As a result: the Paecilomyces gossypii (Paecilomyces heliothis) has effects on 5 strains, wherein the effects on Streptomyces scabies (Streptomyces scabies)4.0076 and Streptomyces albidoflavus (Streptomyces albicans) 4.3598 are the most obvious.
Example 7
And (3) mixed bacterium growth inhibition test: streptomyces scabies (Streptomyces scabies)4.0076, Streptomyces scabies (Streptomyces scabies)4.1765, Streptomyces acidocaldarius (Streptomyces acidiscaibes) 4.1789, Streptomyces albidoflavus (Streptomyces albedo) 4.3598, and Streptomyces european scabies (Streptomyces scabies) each 50ul were spread on the same plate (ISP-2), and 10ul of Paecilomyces gossypii (Paecilomyces heliophilus) was spotted locally on the plate. Culturing at 28 ℃. Observed after 2 days.
As a result: paecilomyces gossypii (Paecilomyces heliothis) has an inhibitory effect on mixed pathogenic bacteria.
Example 8
The co-culture bacteriostasis test method comprises the following steps: streptomyces scabies (Streptomyces scabies)4.0076, Streptomyces scabies (Streptomyces scabies)4.1765, Streptomyces acidocaldarius (Streptomyces acidiscaibes) 4.1789, Streptomyces albidoflavus (Streptomyces albedo) 4.3598, Streptomyces european scabies (Streptomyces scabies) 50ul each and 10ul of Paecilomyces gossypii (Paecilomyces heliothitis) were mixed and spread on ISP-2, and cultured at 28 ℃. Observed after 2 days.
As a result: paecilomyces gossypii (Paecilomyces heliothis) and 5 pathogenic bacteria can grow preferentially in the symbiotic process, and the other three bacteria have no sign of growth.
Claims (10)
1. A strain of Paecilomyces gossypii (Paecilomyces heliothis) is Sanju-18, and the preservation number is CGMCC No. 15869.
2. A microbial agent characterized by comprising the Paecilomyces gossypii strain Sanju-18 according to claim 1 as an active ingredient.
3. The microbial agent according to claim 2, wherein the microbial agent is used in a medicament for preventing and treating potato scab.
4. The use of the paecilomyces gossypii strain Sanju-18 according to claim 1 in a medicament for preventing and treating potato scab.
5. Use according to claim 3 or 4, characterized in that: the potato scab is caused by streptomyces scabrosus.
6. Use according to claim 3 or 4, characterized in that: the potato scab is caused by streptomyces galileoparus.
7. Use according to claim 3 or 4, characterized in that: the potato scab is caused by streptomyces acidoscab.
8. Use according to claim 3 or 4, characterized in that: the potato scab is caused by streptomyces albidoflavus.
9. Use according to claim 3 or 4, characterized in that: the potato scab is caused by streptomyces scab.
10. Use according to claim 3 or 4, characterized in that: the potato scab is caused by streptomyces scab.
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| CN111727963A (en) * | 2019-03-20 | 2020-10-02 | 宁夏大学 | Carrier microbial inoculum compound for preventing and controlling scab and preparation method thereof |
| CN114149949B (en) * | 2021-12-09 | 2023-06-06 | 江西省农业科学院农业应用微生物研究所（江西省农村能源研究中心） | Streptomyces JXJA01 and biological agent and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107034158A (en) * | 2017-04-27 | 2017-08-11 | 中国科学院微生物研究所 | Siam bacillus B 3 and its application |
| CN107432286A (en) * | 2016-05-25 | 2017-12-05 | 中国科学院微生物研究所 | A kind of microbial bacterial agent and its preparation method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107432286A (en) * | 2016-05-25 | 2017-12-05 | 中国科学院微生物研究所 | A kind of microbial bacterial agent and its preparation method and application |
| CN107034158A (en) * | 2017-04-27 | 2017-08-11 | 中国科学院微生物研究所 | Siam bacillus B 3 and its application |
Non-Patent Citations (3)
| Title |
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| Antibacterial, Antifungal and Anticancer Activity of Five Strains of Soil Microorganisms Isolated From Tangkuban Perahu Mountain by Fermentation;Desak Gede Sri Andayani等;《HAYATI Journal of Biosciences》;20160205;第22卷;186-190页 * |
| 孙利军 等.马铃薯土传病害（疮痂病）综合防控试验.《农民致富之友》.2017,(第2017年9期), * |
| 马铃薯土传病害（疮痂病）综合防控试验;孙利军 等;《农民致富之友》;20170930(第2017年9期);112页 * |
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