CN1036307C - A new antibiotic pesticides-Ningnan Meisu - Google Patents
A new antibiotic pesticides-Ningnan Meisu Download PDFInfo
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- CN1036307C CN1036307C CN 93104287 CN93104287A CN1036307C CN 1036307 C CN1036307 C CN 1036307C CN 93104287 CN93104287 CN 93104287 CN 93104287 A CN93104287 A CN 93104287A CN 1036307 C CN1036307 C CN 1036307C
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- antibiotic
- ningnanmycin
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Abstract
The present invention relates to a new antibiotic pesticide-Ningnanmycin, more specifically to a first discovered cytidine peptide antibiotic, namely an antibiotic 16A-6I which is generated from a streptomycete 16A-6 strain, a new variety of streptomyces noursei, namely a streptomyces noursei xichang variety. The antibiotic pesticide is a new pesticide which has the advantages of low toxicity, low residue, no teratogenic, mutagenic and cancerogenic action, no storage problem and no environmental pollution. The relative prevention rate of bacterial blight of rice is about 70% and even as high as 90%, and the rate of the yield increase is 10 to 20% and even as high as 35%. In addition, the Ningnanmycin has conspicuous effects on preventing powdery mildew of wheat, vegetables, flowers, etc. and increasing yield, and has excellent effect on preventing small round sclerotial disease of rice, peacock macula disease of olive, scrab and tobacco mosaic.
Description
The invention belongs to the microbial fermentation engineering technology.
Ningnanmycin belongs to alkaline water dissolubility cytosine nucleoside antibiotic, is mainly used in the white leaf of control paddy rice and picks up disease, also can be used for preventing and treating paddy rice culm rot, wheat powdery mildew, vegetables powdery mildew, tobacco mosaic virus disease etc.The main medicine that is used at present to prevent and treat bacterial blight of rice both at home and abroad is a chemical pesticide, is representative with withered peaceful, the phenazine oxide of leaf.
Since the fifties, because the use of chemical pesticide brings environmental pollution, cause public hazards, therefore, the development of public nuisance-free agricultural chemicals---farm antibiotics is subjected to very big attention in the world, Japan is one of the most successful country of farm antibiotics research, be control rice blast, the rice sheath blight, successively gone into operation " blasticidin-S s ", " Polyoxin ", " kasugarnycin ", " valida ", " griseofulvin ", " cycloheximide " etc., but the white leaf Xian of paddy rice disease etc. is not seen the field experiment report, India, China and some countries of west, also carried out similar research, reported IIerbicidin, Azomultin, Crtromycin, Xanthostiu, more than ten kind of Nogimyein, Prumycin etc. have than the active antibiotic of Johnson ﹠ Johnson's reason the former bacterium of bacterial blight of rice, but, do not see the land for growing field crops application report of preventing and treating bacterial blight of rice owing to poisoning and some other reason yet.
In order to control disease, improve grain yield, environment purification guarantees human and livestock health, seeks efficient, low toxicity, low-residual, does not have the nuisanceless broad-spectrum antibiotic agricultural chemicals of " three cause " and cumulative effect, is main purpose of the present invention.
Technical solution of the present invention is: screening has the antibiotics generated bacterium kind of high physiologically active → classify and identify → carry out the fermentating condition test → antibiotic that produces is separated and structure evaluation → production technology → toxicological test → field experiment producing bacterium to the former bacterium of bacterial blight of rice.
Main contents of the present invention:
1. produce the evaluation of bacterium
Ningnanmycin produces bacterium 16A-6 bacterial strain, according to morphological feature, physio-biochemical characteristics, utilization of carbon source, result of the tests such as cell wall proximate analysis and antagonism: morphological feature: on the Gause I medium, fibrillae of spores is the spacious spiral of pine, under the Electronic Speculum, spore ovalize 0.9 * 1.3 μ m, round diameter 1.2 μ m, spore surface band spinelet; Cultural characteristic: it is luxuriant to grow on the Gao Shi agar medium, the short flannel shape, and aerial mycelium changes light red ash, adularescent secondary hyphae, the light sallow of substrate mycelium, no soluble pigment by dark spider web ash; Physio-biochemical characteristics: on tyrosine agar, do not produce melanin, the starch hydrolysis, gelatin liquefaction, milk peptonizes, nitrate reduction, do not produce hydrogen sulphide, do not grow on the cellulose: utilization of carbon source, can utilize D---glucose, D---fructose, sucrose, mannitol, inositol, do not utilize rhamnose, raffinose, to D---wood sugar, L---arabinose utilizes suspicious; Cell-wall components is the I type; Antagonism: strain cultured solution is to Gram-negative bacteria, minority Gram-positive bacteria, saccharomycete, rice banded sclerotial blight cause of disease bacterium, the rice blast pathogen, olive scab pathogen has inhibitory action, and is strong to former bacterium of bacterial blight of rice and the former bacterium inhibitory action of culm rot.
According to above result, with this bacterial strain and streptomyces noursei relatively, think that streptomycete 16A-6 bacterial strain is a new variant of streptomyces noursei, name and be the mutation of streptomyces noursei Xichang (Streptomyces noursoi var.xichangansisn.var).Preserving number is: CGMCC NO.0194 preservation date: on April 23rd, 1993 depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center (Beijing)
2. the separation and the evaluation of antibiotic Ningnanmycin (16A-6I)
Behind the filtering fermentation liquor, filtrate is by 732
#NII
4 +Behind the type cationic ion-exchange resin, remove the upper strata protein precipitation with salt-free washing, use 0.3N ammoniacal liquor wash-out again, collect phosphotungstic acid reacting positive part, be evaporated to small size, concentrate is by the big net resin of D-110, adsorption chromatography, with no salt solution exhibition layer, collect the phosphotungstic acid reactive moieties, freeze-drying gets white crude product, crude product is through the neutral alumina column chromatography, wash with 10 times of volume 75% methanol aqueous solutions earlier, wash with 25% methanol aqueous solution again, collect phosphotungstic acid reacting positive part, concentrating under reduced pressure, freeze-drying gets white amorphous powder, is further purified through high performance liquid chromatography more at last, gets Ningnanmycin (16A-6I) chromatographically pure sample.
With the white amorphous powder of Ningnanmycin (16A-6I), carry out physicochemical property and detect 195 ℃ of decomposition of fusing point, optical activity [a]
D 25+ 38 °, molecular weight 444[mass spectrometry], molecular formula: C
16II
25N
7O
3, soluble in water, be insoluble in common organic solvents such as alcohol, ketone, ethyl acetate, chloroform, in the pH2-8 phosphate buffer, carry out paper electrophoresis, shift to negative pole, show alkalescence.Chromogenic reaction: ninhydrine, biuret, potassium permanganate, phosphotungstic acid are positive, slope mouth ferric trichloride, anthrone, alpha-Naphthol are negative.
16A-6I is carried out chemical analyses such as ultraviolet spectra, infrared spectrum, nuclear magnetic resoance spectrum and hydrolysate, determine that Ningnanmycin 16A-6I is the gougerotin isomer, difference is to form in the constituent, the former is the L serine, and the latter is the D serine, and antibiotic physiologically active has notable difference, to sum up analyze, the anti-16A-6I of farming is the cytidine peptide type antibiotics of finding first, names to be Ningnanmycin, and its structural formula is:
3. the technological process of production of Ningnanmycin:
Slant strains → first order seed → secondary seed → fermentation tank → zymotic fluid → plate-frame filtering → film concentrates (2-3 doubly) → concentrate and adds 0.1-0.3 Sodium Benzoate → bottling (more than 10000 units per ml) → finished product.
Advantage of the present invention:
1. low toxicity, low-residual, nothing " three cause "
1. acute toxicity experience: Ningnanmycin zymotic fluid freeze drying product are to mouse peritoneal injection L
D50=103.6mg/kg; Oral L
D50=6485mg/kg; Through skin L
D50＞10000mg/kg, rats by intraperitoneal injection L
D50=126.1mg/kg; Oral L
D50=5492mg/kg.
2. fish half tolerance concentration test: Ningnanmycin zymotic fluid freeze drying product are 3323mg/L to crucian half tolerance concentration.
3. subacute toxicity test: low dose group (100mg/kg, be equivalent to 50 times of field dosage) and in dosage group (400mg/kg is equivalent to 200 times of field dosage) and control group relatively, in body weight, ingest, basically identical such as activity, high dose group death midway is because of decreasing on the basis with infection at medicine, the blood picture of survival rats, biochemical indicator check result and control group be no significant difference relatively, according to the boost method with 1/13L
D50Index K＞12.8 are accumulated in (middle dosage group) measuring and calculating, think that this medicine does not have cumulative effect substantially.
4. mutagenicity test: Ningnanmycin does not all have mutagenesis to mouse breastbone marrow micronucleus mutagenesis testing and mouse testis cell chromosome distortion analysis.
5. adopt instruments such as high pressure liquid chromatograph and data processor, detecting standard specimen is the Ningnanmycin chromatographically pure; The examination method rate of recovery is about 98%, to the retention analysis of all kinds of crops sample, beanpod is 0.0160ppm, and the rice leaf is 0.0952ppm, paddy soil is 0.0003ppm, cowpea soil is 0.0034ppm, and cucumber soil, paddy field water either do not detect, millet straw 0.1133ppm, chaff 0.1113ppm, rice 0.0350ppm, wheat 0.0002ppm belongs to low-residual.
2. preventive effect is good, increase yield significantly.Prove according to 5 provinces, 3884 mu of field experiments in 6 counties, Ningnanmycin to withered peaceful, the phenazine oxide of bacterial blight of rice and leaf, enemy withered two etc. relatively, the Ningnanmycin relative control effect is generally 70%, high reaches more than 90%, average 85.41%, effect of increasing production is generally 10～20%, is up to 35%, two average preventive effects are 62.14%～75.63% and withered peaceful, the phenazine oxide of leaf, enemy are withered, and effect of increasing production is about 10%.
3. raw material is easy to get, inexpensive, fermentation titer is stable, is fit to large-scale industrial production, production equipment can with other production of antibiotics Device-Generals.
4. to light, heat, sour more stable, concentrate is preserved at normal temperatures and was not lost efficacy in 2 years, to the people, animal nonhazardous, crop there is not poisoning, no off-odor, not chafe, resistance of rainwater washing against, in the simultaneous field of damage by disease and insect, can with Rogor, decamethrin is used with, not only can reach the control positive effect, and economize the labour, save time, thereby reduced drug cost.
Embodiment: 1
1989, Chengdu factory carry out 1 ton of test tank and 6 tons produce jar totally 20 batches production investigate, prove that this bacterial strain produces to tire to be stabilized in 5000 units per ml that the product Ningnanmycin is stored at normal temperatures and do not lost efficacy in 2 years.Crossed and generally take place 3580 mu of rice field demonstration tests of bacterial leaf-blight by water logging in the Sichuan county with this antibiotic, though the chance rain of spray medicine, do not subtract drug effect yet, control efficiency average out to 86.48%, the withered peaceful preventive effect of leaf of contrast then is 77.14%, and phenazine oxide is 66.85%, effect of increasing production: Ningnanmycin is 11.77%, leaf is withered rather to be 9.09%, and the leaf three-bristle cudrania is 5.35% only.
Embodiment: 2
Ningnanmycin and triadimefon, sulphur colloidal suspension agent compared wheat powdery mildew in certain two county prevent and treat result of the test, the Ningnanmycin preventive effect is 80.19%, effect of increasing production is 27.73%, and the control efficiency of triadimefon next at similarity condition be 66.34%, effect of increasing production is 26.05%, the control efficiency of sulphur colloidal suspension agent is 67.54%, and effect of increasing production is 16.81%.
Embodiment: 3
With 4000 times of the triadimefons of Ningnanmycin (concentration 50ppm/ml) and concentration 15%, prevent and treat vegetables powdery mildew tests such as cucumber, cowpea with 1000 times of concentration 70% thiophanate methyls, the result, the Ningnanmycin preventive effect: cucumber is 85.7%, cowpea is 92.8%, effect of increasing production: cucumber is 37.3%; Triadimefon is 81.0% to the cucumber preventive effect, is 90.3% to cowpea, and effect of increasing production is 36.6%; Thiophanate methyl is 64.3% to the cucumber preventive effect, and cowpea is 78.9%, and effect of increasing production is 9.2%.
Embodiment: 4
Continuous 2 years of 1990-1991 prevents and treats test with first south mycin to tobacco mosaic in the somewhere, if 83 to increase anti-agent be contrast agents, clear water is used conventional spray for contrast, mu dosage (specification 1%) 750 milliliters, 83 increase 750 milliliters of anti-agent, convert 75 kilograms of sprayings of water, dispenser totally three times, the result shows, the absolute preventive effect of Ningnanmycin is 87.38%, and effect of increasing production is 23.2%; 83 preventive effects that increase anti-agent only are 22.47%, and Ningnanmycin obviously is better than 83 and increases anti-agent.
Claims (3)
1. the antibiotic agricultural chemicals of producing with microbial fermentation technology is characterized in that; It produces bacterium 16A-6 bacterial strain is a new variant of streptomyces noursei, names to be the mutation of streptomyces noursei Xichang (Streptomyces noursei varxichangensis nvar), and preserving number is: CGMCC NO.0194; Its molecular weight is the 444[mass spectrometry],
Molecular formula is C
16II
25N
7O
8, structural formula is:
2. the antibiotic agricultural chemicals of producing with microbial fermentation technology according to claim 1, it is characterized in that: hydrolysate is a cytidine peptide type antibiotic, and its serine is the L-serine.
3. the antibiotic agricultural chemicals of producing with microbial fermentation technology according to claim 2, feature is altogether: its fusing point is 195 ℃, and optical activity is [a]
D 25+ 38 °.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 93104287 CN1036307C (en) | 1993-04-23 | 1993-04-23 | A new antibiotic pesticides-Ningnan Meisu |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 93104287 CN1036307C (en) | 1993-04-23 | 1993-04-23 | A new antibiotic pesticides-Ningnan Meisu |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1093869A CN1093869A (en) | 1994-10-26 |
| CN1036307C true CN1036307C (en) | 1997-11-05 |
Family
ID=4985113
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 93104287 Expired - Lifetime CN1036307C (en) | 1993-04-23 | 1993-04-23 | A new antibiotic pesticides-Ningnan Meisu |
Country Status (1)
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| CN (1) | CN1036307C (en) |
Cited By (1)
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|---|---|---|---|---|
| US9192166B2 (en) | 2013-02-11 | 2015-11-24 | Bayer Cropscience Lp | Compositions comprising a streptomyces-based biological control agent and an insecticide |
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| CN1050846C (en) * | 1995-08-03 | 2000-03-29 | 中国医学科学院医药生物技术研究所 | Antitumor antibiotic-Yunnan-mycin |
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| CN101978827B (en) * | 2010-12-01 | 2012-11-28 | 德强生物股份有限公司 | Ningnanmycin composition and preparation method thereof |
| CN102067855B (en) * | 2011-02-17 | 2013-04-10 | 德强生物股份有限公司 | Polyoxin and ningnan mycin compounded pesticide |
| CN102160549B (en) * | 2011-02-17 | 2013-04-10 | 德强生物股份有限公司 | Compound pesticide containing cyprodinil and ningnanmycin |
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| CN102363753B (en) * | 2011-03-07 | 2012-11-21 | 浙江师范大学 | Streptomyces termitum ACT-2 strain, culture and application thereof |
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| CN107549181A (en) * | 2017-09-30 | 2018-01-09 | 京博农化科技股份有限公司 | A kind of composition containing Ningnanmycin |
| CN107573403B (en) * | 2017-10-17 | 2020-07-28 | 福州奥尼多生物科技有限公司 | Method for purifying ningnanmycin |
| CN109699665B (en) * | 2018-12-19 | 2021-03-12 | 深圳诺普信农化股份有限公司 | Seed coating agent containing captan and application thereof |
| CN111411050B (en) * | 2019-01-04 | 2023-03-17 | 西北农林科技大学 | F01 actinomycete and antiviral application thereof |
| CN113337553B (en) * | 2019-05-31 | 2022-10-21 | 中国医学科学院医药生物技术研究所 | Herbicidin biosynthesis gene cluster and application thereof |
-
1993
- 1993-04-23 CN CN 93104287 patent/CN1036307C/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9192166B2 (en) | 2013-02-11 | 2015-11-24 | Bayer Cropscience Lp | Compositions comprising a streptomyces-based biological control agent and an insecticide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1093869A (en) | 1994-10-26 |
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