CN1237169C - Bactericide bandung mycin and its upplication - Google Patents
Bactericide bandung mycin and its upplication Download PDFInfo
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- CN1237169C CN1237169C CN 03140091 CN03140091A CN1237169C CN 1237169 C CN1237169 C CN 1237169C CN 03140091 CN03140091 CN 03140091 CN 03140091 A CN03140091 A CN 03140091A CN 1237169 C CN1237169 C CN 1237169C
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- bandung
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Abstract
The present invention relates to a bactericide, namely Wanlongmycin and application thereof. The bactericide has an effective component molecular formula: C53H67N13O13S2. The molecular weight of the bactericide is 1157, and a chemical structural formula of the bactericide is given on the right part of a specification. The bactericide is suitable for preventing and treating bacterium and oomycetes diseases which comprise bacterial wilt, rice bacterial leaf streak, rice bacterial blight, bacterial soft rot, plant epidemic diseases, downy mildew, downy epidemic mildew and late blight, and the bactericide is effective for diseases caused by fusarium, hay bacillus and colibacillus.
Description
Technical field
The present invention relates to plant protection and technical field of pesticide, specifically be meant a kind of sterilant bandung mycin and application thereof.
Background technology
Agricultural antibiotic has characteristics such as selectivity is strong, efficient, low toxicity, and the research and development microbiotic is the focus of pesticide research.Though at present the microbiotic kind of the controlling plant diseases of report is many, can be on producing the kind of large scale application less, still lack prevent and treat disease efficiently, the agricultural antibiotic of low toxicity.The oomycetes disease is meant the disease that is caused by main cash crop such as Oomycete algae shape fungal infection vegetables, fruit tree, common as melon eqpidemic disease, capsicum epidemic disease, tomato late blight, the late blight of potato, downy mildew of crucifers, downy mildew of cucurbits, downy mildew of lichee etc., these disease PD are rapid, endanger very greatly, usually cause big area to destroy, lose and receive.To the control of plant Oomycete disease, mainly rely on chemical prevention at present, common fungicide mainly is the medicines such as metalaxyl, Sandofan and home-made metaxanin of import, and these medicaments cost an arm and a leg, and uses for many years, develops immunity to drugs in some areas.
Summary of the invention
Purpose of the present invention is exactly in order to solve above-mentioned the deficiencies in the prior art part, and a kind of sterilant bandung mycin and application thereof are provided.This sterilant is produced by a new streptomyces bacterial classification, and it is applicable to controlling plant diseases etc., the fungus-caused disease of bacterium and Oomycete particularly, and also free from environmental pollution, have attracting prevention effect.
The present invention at first provides the bacterial strain of a new streptomyces, described bacterial strain is named as Streptomyces griseovariabilis subsp.Bandungensis n.subsp.GAAS2507, it is separating obtained from pedotheque, described bacterial strain is being positioned at Chinese typical culture collection center (CCTCC) preservation in Wuhan on July 7th, 2003, and deposit number is CCTCC NO:M203059.
A kind of sterilant bandung mycin of the present invention is characterized in that the molecular formula of its effective constituent is: C
53H
67N
13O
13S
2, molecular weight is: 1157, and chemical structural formula is:
A kind of sterilant bandung mycin of the present invention, be applicable to control bacterium and oomycetes disease, comprise bacterial wilt, bacterial leaf streak of rice, bacterial blight of rice, bacterial soft rot and vegetable plague, oidium, peronospora tabacina, late blight, effective to the microbial disease of reaping hook and Bacillus subtilus and intestinal bacteria.
A kind of sterilant bandung mycin of the present invention can prepare by following method:
The first step inoculates the culture of GAAS2507 into the nutrition substratum, and growth is 36 hours under the vibration of 32 ℃ and 250rpm;
Second step was got sophisticated fs nutrition culture inoculating nutrient medium, obtained secondary nutrition culture, cultivated 48 hours under 32 ℃ and 300rpm condition, and sophisticated strain fermentation is cultivated, and kept cultivating 4 days under 32 ℃ and 300rpm; The sophisticated fermented product of available suitable solvent extraction, and pass through chromatographic separation and obtain metabolite.
Culture feature such as the following table of bacterial strain GAAS2507 of the present invention:
Table 1
| Substratum | The base silk | The gas silk | Pigment |
| Gause I | Smoky gray | Colourless or yellow-white | Do not have |
| Cha Shi sucrose | Canescence | Colourless or little ash | Do not have |
| No. one, Ke Shi | Shallow smoky gray | Colourless to yellow-white | Do not have |
| The glucose asparagine | White (on a small quantity) | Do not have | Do not have |
| The glycerine asparagine | Almost do not have | Light brown | The parrot hat is yellow |
| Potato ball | Light gray or cigarette ash | Cigarette ash | Chocolate |
| Oatmeal | Dark grey | Nothing or greyish white | Do not have |
| Tyrosine | Do not have | Light gray | Brown |
| Inorganic salt starch | Light gray | Do not have | Do not have |
| Glucose yeast | Cigarette ash | White to yellowish-white | Do not have |
| Calcium malate | Greyish white | Greyish white | The jasmine Huang |
The morphological specificity of bacterial strain GAAS2507 of the present invention is: 2507 fibrillaes of spores that produce bacterium are loose volution, and spore surface is smooth.On the Gause I substratum, its gas silk is smoky gray, and the base silk is colourless to Huang Zhese, does not produce pigment.Similar to gray variant streptomycete on form, cultural characteristic and physio-biochemical characteristics, there is significant difference again in correction, therefore names to be gray variant streptomycete Bandung subspecies (Streptomyces griseovariabilis subsp.Bandungensis n.subsp.).
The physiological characteristic of bacterial strain GAAS2507 of the present invention is: gelatine liquefication is suspicious; Milk solidifies and peptonizes strongly; The hydrolyzed starch ability is stronger; Do not grow on the Mierocrystalline cellulose; Do not produce H
2S.Can utilize glucose, lactose, raffinose, maltose, N.F,USP MANNITOL, inositol, rhamnosyl, seminose, semi-lactosi; Do not utilize D-fructose, sorbose, pectinose, wood sugar, sucrose, synanthrin, sweet and pure, sorbyl alcohol.The present invention compared with prior art has following advantage and beneficial effect:
1. simple, easy to operate, the easy preparation of preparation technology of the present invention.
2. sterilant bandung mycin of the present invention is effective to control of plant disease such as oomycetes agent bacteriums, lacks than the conventional dose consumption, can overcome the resistance of Plant diseases to conventional pesticide.
3. sterilant of the present invention is safe in utilization, free from environmental pollution, is a kind of non-harmful biological pesticide.
Description of drawings
Fig. 1 is the form synoptic diagram of bacterial strain GAAS2507 of the present invention;
Fig. 2 is the joint cucurbit wilt bacterium bacterium colony synoptic diagram that is suppressed by bandung mycin in the embodiment of the invention four.
Bacterial strain Streptomyces griseovariabilis subsp.Bandungensis n.subsp.GAAS2507 of the present invention is being positioned at Chinese Typical Representative culture collection center (CCTCC) preservation in Wuhan on July 7th, 2003, and deposit number is CCTCC NO:M203059.
Embodiment
Below in conjunction with drawings and Examples, the present invention is done detailed description further.
Embodiment one
Prepare the sterilant bandung mycin by in fermention medium, cultivating bacterial strain GAAS2507.The refrigerated GAAS2507 nutritional medium of 1ml is melted, and the 100ml nutritional medium in the 500ml triangular flask is gone in inoculation, and growth is 48 hours under the vibration of 32 ℃ and 250rpm.
Table 2 nutritional medium composition
| The composition title | Content (%) |
| Glucose corn steep liquor soybean cake powder starch NaCl (NH 4) 2SO 4 CaCO 3 pH | 3 1.5 1.5 1 0.5 0.5 0.5 7.2 |
Get the secondary nutrition culture of nutritional medium that the sophisticated fs nutrition of 3500ml culture is used to inoculate the 35L of 50L stirred-tank fermenter, under 32 ℃ and 300rpm condition, cultivated 48 hours, in the stirred-tank fermenter of 500L, fermentation culture keeps cultivating 4 days under 32 ℃, 300rpm with sophisticated bacterial classification inoculation.The sophisticated fermented product of available suitable solvent extraction, and pass through chromatographic separation and obtain metabolite---the sterilant bandung mycin.
Embodiment two
The present invention can be by test broth extract and the preparation of the bandung mycin of tracking sterilization agent is during fermentation for example analyzed fermentation broth extract by bioassay method.
Fermentation finishes, fermented product from 500L, mycelia is obtained in method separation with filter press, the acetone soln of mycelia with 1: 2 leached 10 hours, leaching liquid is concentrated remove acetone, use the ethyl acetate extraction working substance again, the acetic acid ethyl acetate extract concentrating under reduced pressure is obtained oily matter, the extraction concentrated solution is mixed silica gel and is crossed post, uses repeatedly silica gel column chromatography and concentrated of ethyl acetate, sherwood oil, obtains the pure product of oily mater.
Embodiment three
A kind of sterilant bandung mycin of the present invention is applicable to control bacterium and oomycetes disease, is included in the field and uses the germ amount of suppression.Come the effect of controlling plant diseases relevant with access times and using method by the sterilant bandung mycin, relevant with the kind of the disease of being prevented and treated simultaneously.The present invention realizes using preparation by the disclosed technical scheme of Chinese invention patent application (number of patent application is 99117029.6).
Sterilant bandung mycin acetone soln with 2 times of dilutions of sterilized water, is adopted inhibition curve and the MIC of double-deck agar cup-plate method mensuration to Bacillus subtilus and bacterial stripe bacterium.Use the result shown in table 3,4.
Table 3 sterilant bandung mycin is to the restraining effect of Bacillus subtilus
| Concentration (μ g/ml) | Inhibition zone radius (mm) | ||||
| 1 | 2 | 3 | 4 | On average * | |
| 100 50 25 12.5 6.25 3.125 1.563 0.781 0.391 0.195 CK (sterilized water) | 8.25 8.00 7.10 6.40 5.50 4.75 4.25 3.25 2.75 2.50 2.50 | 8.50 8.25 7.10 6.50 6.25 5.00 4.40 3.50 2.90 2.50 2.50 | 10.50 9.75 8.00 7.00 6.10 5.00 4.25 3.50 2.75 2.50 2.50 | 11.50 9.75 8.75 7.75 6.40 5.10 5.50 3.90 3.00 2.50 2.50 | 9.688±0.786A 8.938±0.472A 7.738±0.399B 6.913±0.308BC 6.063±0.197C 4.963±0.075D 4.600±0.302D 3.538±0.134E 2.850±0.061E 2.500±0.000E 2.500±0.000E |
| Regression equation ** Y＝0.0288484890X-0.4820479049 | |||||
*: through the DUNCANShi of SAS statistical software multiple comparisons, R=0.938006, P=0.0001.
*: according to the concentration logarithm Y of sterilant bandung mycin and the square value X of inhibition zone radius, with the SAS statistical software try to achieve regression equation (R=0.940579, P=0.0001).
Table 4 sterilant bandung mycin is to the restraining effect of xanthomonas oryzae pv. oryzicola
| Concentration (μ g/ml) | Inhibition zone radius (mm) | |||||
| 1 | 2 | 3 | 4 | 5 | On average * | |
| 50 25 12.5 6.25 3.125 1.563 0.781 | 7.25 7.00 4.50 5.25 3.50 3.00 2.75 | 7.25 4.75 5.00 4.75 3.50 3.00 2.75 | 6.50 5.50 4.25 4.00 3.50 3.00 2.75 | 7.75 6.50 4.50 3.50 3.25 3.00 2.75 | 8.00 7.25 4.50 4.00 3.25 3.00 2.75 | 7.35±0.257A 6.20±0.470B 4.55±0.122C 4.30±0.310C 3.40±0.061D 3.00±0D 12.75±0D |
| CK (sterilized water) regression equation ** Y＝0.0348525860X-0.0003572797 |
*: through the DUNCANShi of SAS statistical software multiple comparisons, R=0.916090, P=0.0001.
*: according to the concentration logarithm Y of sterilant bandung mycin and the square value X of inhibition zone radius, with the SAS statistical software try to achieve regression equation (R=0.857593, P=0.0027).
Embodiment four
Adopt medium liquid dilution method and seed agar piece method to measure inhibition curve and the LD of cucumber parasitica, joint cucurbit wilt bacterium, rice blast fungus and mango charcoal myiasis bacterium
50, use the result as shown in table 5.
Experimental result shows that the sterilant bandung mycin has obvious suppression effect (seeing Table 5) to cucumber parasitica mycelial growth, and when the concentration of 2507A1 was 50 μ g/ml, cucumber parasitica mycelial growth was suppressed fully.
Try to achieve the LD of sterilant bandung mycin by regression equation to cucumber parasitica mycelial growth
50=1.256 μ g/ml.
Table 5 sterilant bandung mycin is to the restraining effect effect of cucumber parasitica growth
| Concentration (μ g/ml) | The mycelial growth diameter * (42hr mm | Mycelial growth inhibition rate (%) | Antibacterial work probability value |
| 50 25 12.5 6.25 3.125 1.563 0.781 CK (sterilized water) | 0±0F 8.7±0.539E 12.5±0.880D 14.0±0.158CD 14.5±0.447C 16.0±1.007C 26.0±0.447B 40.9±0.843A | 100 78.7 69.4 65.8 64.5 60.9 36.4 - | - 5.7961 5.5072 5.4070 5.3719 5.2767 4.6505 |
| Regression equation ** Y＝0.612791330X+4.939325911 | |||
*: through the DUNCANShi of SAS statistical software multiple comparisons, R=0.987862, P=0.0001.
*: according to the concentration logarithm X of antibacterial work probability value Y and sterilant bandung mycin (the 6 groups of data in back), with the SAS statistical software try to achieve regression equation (R=0.826143, P=0.0121).
The result shows that the sterilant bandung mycin also has good inhibitory effect (to see Table 6, Fig. 1) to joint cucurbit wilt bacterium mycelial growth.Try to achieve the LD of sterilant bandung mycin according to regression equation to joint cucurbit wilt bacterium mycelial growth
50=20.570 μ g/ml.
Table 6 sterilant bandung mycin is to the restraining effect effect of joint cucurbit wilt bacteria growing
| Concentration (μ g/ml) | The mycelial growth diameter * (60hr mm) | Mycelial growth inhibition rate (%) | Antibacterial work probability value |
| 50 25 12.5 6.25 3.125 1.563 0.781 0.391 0.195 CK (sterilized water) | 15.6±1.317F 16.7±0.436F 19.8±1.114E 23.4±0.030D 24.6±0.678D 27.8±1.800C 32.8±0.200B 34.3±0.200AB 33.4±0.748AB 36.0±1.049A | 56.7 53.6 45.0 35.0 31.7 22.8 8.9 4.7 7.2 - | 5.1687 5.0904 4.8743 4.6147 4.5239 4.2546 3.6531 3.3253 3.5462 |
| Regression equation ** Y＝0.807660378X+3.939351467 | |||
*: through the DUNCANShi of SAS statistical software multiple comparisons, R=0.939921, P=0.0001.
*: according to the concentration logarithm X of antibacterial work probability value Y and sterilant bandung mycin, with the SAS statistical software try to achieve regression equation (R=0.937433, P=0.0001).
Find out that from test-results sterilant bandung mycin effect of the present invention is fine, and is not only fine to the bacterial diseases of plants prevention effect, and also fine to plant Oomycete disease effect, environment is not polluted.Simultaneously, can overcome the resistance of phytopathogen to medicament.As the control agent of Plant diseases, will help the stable yields and the high yield of agricultural, thereby produce bigger economic and social benefit.
As mentioned above, can realize the present invention preferably.
Claims (4)
1. the bacterial strain of a streptomyces, it is characterized in that, it is separating obtained from pedotheque, described bacterial strain is Streptomyces griseovariabilis subsp.Bandungensis n.subsp.GAAS2507, in China's typical culture collection center preservation, deposit number is CCTCC NO:M 203059.
2. a sterilant bandung mycin is characterized in that, it produces by cultivating the described bacterial strain of claim 1, and its molecular formula is: C
53H
67N
13O
13S
2, molecular weight is: 1157, and chemical structural formula is:
3. the application of the described sterilant bandung mycin of claim 2 is characterized in that, described sterilant bandung mycin is used to prevent and treat vegetative bacteria and oomycetes disease.
4. according to the application of the described sterilant bandung mycin of claim 3, wherein said vegetative bacteria and oomycetes disease are bacterial wilt, bacterial leaf streak of rice, bacterial blight of rice, bacterial soft rot, the phytobacterial disease that vegetable plague, oidium, peronospora tabacina, late blight or sickle-like bacteria, Bacillus subtilus, intestinal bacteria cause.
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|---|---|---|---|
| CN 03140091 CN1237169C (en) | 2003-08-06 | 2003-08-06 | Bactericide bandung mycin and its upplication |
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| CN1237169C true CN1237169C (en) | 2006-01-18 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100497595C (en) * | 2006-09-21 | 2009-06-10 | 广东省农业科学院植物保护研究所 | Extract of soil actinomycetes fermenting liquid and its preparation and application |
| CN101928685A (en) * | 2010-03-11 | 2010-12-29 | 王家琛 | Streptomyces albus MC-15 bacterial strain as well as method and application thereof for preparing fermentation liquor thereby |
| CN102363753B (en) * | 2011-03-07 | 2012-11-21 | 浙江师范大学 | Streptomyces termitum ACT-2 strain, culture and application thereof |
| CN102696682B (en) * | 2012-04-20 | 2014-12-03 | 广东省农业科学院植物保护研究所 | Wanlongmycin dimethomorph suspending agent, preparation method thereof and application |
| CN111454869B (en) * | 2020-06-22 | 2020-10-09 | 滨州医学院 | Marine streptomyces and application thereof |
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