CN103088068A - Preparation method and application of soil rare actinomyces broth - Google Patents
Preparation method and application of soil rare actinomyces broth Download PDFInfo
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- CN103088068A CN103088068A CN2011103448374A CN201110344837A CN103088068A CN 103088068 A CN103088068 A CN 103088068A CN 2011103448374 A CN2011103448374 A CN 2011103448374A CN 201110344837 A CN201110344837 A CN 201110344837A CN 103088068 A CN103088068 A CN 103088068A
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- soil rare
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- rare actinomycete
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Abstract
The invention relates to a preparation method of soil rare actinomyces broth, and an application of the broth in preparing anti-fungal pesticides. The soil rare actinomyces broth is obtained by a seed culture of the soil rare actinomyces, a liquid fermentation culture of the soil rare actinomyces, and purification of the broth. According to the invention, a fermentation technology and a purification technology are simple; and the broth has relatively good inhibition effects on five tested pathogenic fungi of wheat scab, apple canker, tomato early blight, pear Botrytis cinerea, and cotton fusarium wilt, can be prepared into biopesticides, and has relatively high economical and social benefits.
Description
Technical field
The present invention relates to a kind of preparation method of soil rare actinomycete fermented liquid, and the application of this fermented liquid in preparation fungicide process.
Background technology
Plant pathogenic fungi is the main pathogenic microbes monoid that harms the crops, and produces to have caused massive losses, utilizes the problems such as pesticide residue that chemical prevention causes and contaminate environment to attract widespread attention.Agricultural chemicals plays very important effect to the development of agricultural, but the development along with society, follow safety and the environmental problem that chemical pesticide causes also day by day to show especially, in recent years, endophyte of plant has caused widely as a kind of new Biological resources to be paid close attention to, seeking from endophyte and found the active compound that some are new, is a feasible approach so separate Substance from the meta-bolites of rare actinomycete.
Agricultural antibiotic is by the secondary metabolite that produces in fermentation process, is the quasi-microorganism agricultural chemicals that the disease that can suppress or kill crop when lower concentration, worm, crop smothering and regulating crop growth are grown.Initiative novel microorganism source pesticide control plant pest has very large market outlook and using value, thereby the actinomycetes resource that screening has a high fungicidal activity has great practice significance to the exploitation novel agrochemical.Up to now, the microbiotic of having found in the world is nearly 2nd/3rd, is produced by actinomycetes, and the Antibiotics that wherein produces with streptomycete is maximum.Yet along with finding that microbiotic is more and more, find that from streptomycete new antibiotic probability is more and more less, people begin to seek new antibiotic from rare actinomycete.Rare actinomycete becomes more and more important as the effect in new antibiotic source.
Summary of the invention
The object of the present invention is to provide a kind of soil rare actinomycete fermented liquid that the potential source biomolecule agricultural chemicals is worth that has, with and preparation method thereof, and provide this soil rare actinomycete fermented liquid to fungi, prevention effect preferably to be arranged.
The present invention's microorganism strains used is that the Guangdong Province soil sample is separated and obtained soil microorganisms, and combining form and physiological and biochemical property are initially identified as the Nocardia bacteria 2-200 of Actinomy cetaceae Nocardia.
Purpose of the present invention is achieved through the following technical solutions: a kind of preparation method of soil rare actinomycete fermented liquid comprises the following steps:
(1) seed culture of soil rare actinomycete
With the aseptic inoculation ring with the spore inoculating of bacterial strain on Gao Shi I medium slant, " Z " word method was cultivated under 28 ℃ 5～8 days;
(2) liquid fermentation and culture of soil rare actinomycete
Be inoculated in the 250mL triangular flask that the 100mL fermention medium is housed with aseptic inoculation ring picking 3～5 rings, 25～30 ℃, under 250～300r/min condition, shaking culture is 3～6 days, takes out it is leached supernatant liquor with filter paper, obtains fermented liquid;
(3) purifying of fermented liquid
Fermented liquid is moved in the centrifuge tube of 50mL volume, the centrifugal 10～15min of the whizzer of 3000～5000r/min keeps filtrate, is soil rare actinomycete fermented liquid.
Described Gao Shi I substratum is Zulkovsky starch 20g, KNO
31g, K
2HPO
40.5g, MgSO
47H
2O 0.5g, NaCl 0.05g FeSO
47H
2O 0.01g, agar 20g, pH=7.4～7.6.
Described liquid fermentation medium is glucose 5.0～20g, peptone 1.5～5.0g, NaCl 0.5～5.0g, millet 5.0～20g, CaCO
32.0～4.0g, water 1000mL.
A kind of soil rare actinomycete fermented liquid is prepared from by above-mentioned preparation method.
The application of above-mentioned soil rare actinomycete fermented liquid in preparation fungicide process.
Fermented liquid of the present invention can have inhibition preferably to wheat scab (Fusarium graminearum), canker of apple fruit (Valsa mali Miyabe et Yamada), early blight of tomato (Ahernaria solani), pears gray mold (Botrytis cinerea), cotton wilt (Fusarium oxysporum f.sp.Vasinfectum).Fermented liquid of the present invention can be used for preparing mycotic agricultural chemicals.
The present invention compared with prior art has the following advantages and effect: zymotechnique of the present invention and purifying process are simple.Fermented liquid of the present invention has restraining effect preferably to wheat scab, canker of apple fruit, early blight of tomato, pears gray mold, these five kinds confession examination pathogenic bacterias of cotton wilt, is expected to make biological pesticide, has higher economic and social benefit.
Specific implementation method
Embodiment 1
The preparation method of fermented liquid of the present invention:
(1) seed culture of soil rare actinomycete
With the aseptic inoculation ring with the spore inoculating of bacterial strain on Gao Shi I medium slant, " Z " word method was cultivated 6 days under 28 ℃;
(2) liquid fermentation and culture of soil rare actinomycete
Be inoculated in the 250mL triangular flask that the 100mL fermention medium is housed with aseptic inoculation ring picking 3 rings, 25 ℃, under the 300r/min condition, shaking culture is 4 days, takes out it is leached supernatant liquor with filter paper, obtains fermented liquid;
(3) purifying of fermented liquid
Fermented liquid is moved in the centrifuge tube of 50mL volume, the centrifugal 10min of the whizzer of 5000r/min keeps filtrate, is soil rare actinomycete fermented liquid.
Embodiment 2
Fermented liquid of the present invention suppresses wheat scab, canker of apple fruit, early blight of tomato, pears gray mold, cotton wilt experiment
The fermented liquid that embodiment 1 is obtained is as giving birth to the test sample product, take wheat scab, canker of apple fruit, early blight of tomato, gray mold, cotton wilt as for the examination bacterium.Fermented liquid and PDA substratum are mixed take the ratio of 1: 9 dull and stereotypedly access respectively bacterium cake diameter for the examination bacterium as 5mm in central authorities, and process in contrast with sterilized water, cultivate 3d, adopt the right-angled intersection method to measure antibacterial circle diameter, each is processed and repeats 3 times.The PDA medium component is potato 200g, glucose 20g, agar 15-20g, water 1000ml.Experimental result sees Table 1.
Inhibiting rate (%) the mean number form of table 1 actinomycetes to five kinds of fungies
Claims (5)
1. the preparation method of a soil rare actinomycete fermented liquid is characterized in that comprising the following steps:
(1) seed culture of soil rare actinomycete
With the aseptic inoculation ring with the spore inoculating of bacterial strain on Gao Shi I medium slant, " Z " word method, cultivated under 28 ℃ 5～8 days:
(2) liquid fermentation and culture of soil rare actinomycete
Be inoculated in the 250mL triangular flask that the 100mL fermention medium is housed with aseptic inoculation ring picking 3～5 rings, 25～30 ℃, under 250～300r/min condition, shaking culture is 3～6 days, takes out it is leached supernatant liquor with filter paper, obtains fermented liquid;
(3) purifying of fermented liquid
Fermented liquid is moved in the centrifuge tube of 50mL volume, the centrifugal 10～15min of the whizzer of 3000～5000r/min keeps filtrate, is soil rare actinomycete fermented liquid.
2. the preparation method of a kind of soil rare actinomycete fermented liquid according to claim 1, it is characterized in that: described bacterial strain is that the soil rare actinomycete is the Nocardia bacteria 2-200 of Actinomy cetaceae Nocardia.
3. the preparation method of a kind of soil rare actinomycete fermented liquid according to claim 1, it is characterized in that: described fermention medium is glucose 5.0～20.0g, peptone 1.5～5.0g, NaCl 0.5～5.0g, millet 5.0～20g, CaCO
32.0～4.0g, water 1000mL.
4. a soil rare actinomycete fermented liquid is prepared from by preparation method claimed in claim 1.
5. soil rare actinomycete fermented liquid claimed in claim 4 has effect preferably preventing and treating mycosis, can prepare antimycotic agricultural chemicals.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011103448374A CN103088068A (en) | 2011-11-04 | 2011-11-04 | Preparation method and application of soil rare actinomyces broth |
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|---|---|---|---|
| CN2011103448374A CN103088068A (en) | 2011-11-04 | 2011-11-04 | Preparation method and application of soil rare actinomyces broth |
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|---|---|
| CN103088068A true CN103088068A (en) | 2013-05-08 |
Family
ID=48201196
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| CN2011103448374A Pending CN103088068A (en) | 2011-11-04 | 2011-11-04 | Preparation method and application of soil rare actinomyces broth |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS623789A (en) * | 1985-06-29 | 1987-01-09 | Banyu Pharmaceut Co Ltd | Antibiotic substance tokimycin a and production thereof |
| CN87102770A (en) * | 1986-04-11 | 1987-12-16 | 伊莱利利公司 | Antibiotic A 42125 and production method thereof |
| CN1461807A (en) * | 2002-05-31 | 2003-12-17 | 中国科学院沈阳应用生态研究所 | Fermentation method of producing antifungus active substance using ocean streptomycete |
| JP2006296419A (en) * | 2005-03-04 | 2006-11-02 | Hiroshima Univ | Method for producing antibiotic-producing microorganism by gene disruption and the resultant antibiotic-producing microorganism, and method for producing antibiotic metabolic intermediate |
| CN101165169B (en) * | 2007-09-28 | 2010-06-16 | 厦门大学 | Control method for biological flocculant XM-11 fermentation process grading oxygen supply |
-
2011
- 2011-11-04 CN CN2011103448374A patent/CN103088068A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS623789A (en) * | 1985-06-29 | 1987-01-09 | Banyu Pharmaceut Co Ltd | Antibiotic substance tokimycin a and production thereof |
| CN87102770A (en) * | 1986-04-11 | 1987-12-16 | 伊莱利利公司 | Antibiotic A 42125 and production method thereof |
| CN1461807A (en) * | 2002-05-31 | 2003-12-17 | 中国科学院沈阳应用生态研究所 | Fermentation method of producing antifungus active substance using ocean streptomycete |
| JP2006296419A (en) * | 2005-03-04 | 2006-11-02 | Hiroshima Univ | Method for producing antibiotic-producing microorganism by gene disruption and the resultant antibiotic-producing microorganism, and method for producing antibiotic metabolic intermediate |
| CN101165169B (en) * | 2007-09-28 | 2010-06-16 | 厦门大学 | Control method for biological flocculant XM-11 fermentation process grading oxygen supply |
Non-Patent Citations (2)
| Title |
|---|
| 《农药学学报》 20071231 黄路枝等 "土壤稀有放线菌的选择性分离及其抗菌活性研究" 第59-65页 第9卷, 第1期 * |
| 黄路枝等: ""土壤稀有放线菌的选择性分离及其抗菌活性研究"", 《农药学学报》 * |
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Application publication date: 20130508 |