CN107935880B - Separation and identification of aspergillus fumigatus antioxidant product for improving drought resistance of rice and application thereof - Google Patents
Separation and identification of aspergillus fumigatus antioxidant product for improving drought resistance of rice and application thereof Download PDFInfo
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C233/00—Carboxylic acid amides
- C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C233/16—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms
- C07C233/17—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
- C07C233/18—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by singly-bound oxygen atoms with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/18—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing the group —CO—N<, e.g. carboxylic acid amides or imides; Thio analogues thereof
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- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
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Abstract
The invention discloses separation and identification of an antioxidant product of aspergillus fumigatus and application thereof in improving drought resistance of rice. The Aspergillus fumigatus is deposited under the name ofAspergillus fumigatusSG 17; the preservation unit: china center for type culture Collection; the preservation address is China, Wuhan and Wuhan university; the preservation date is as follows: 5 months and 11 days 2015; the preservation number is CCTCC NO: m2015286. The antioxidant product is (Z) -N- (4-hydroxystyryl) formamide. Experiments prove that the NFA can reduce the contents of proline and malondialdehyde in drought rice, effectively assist the rice in resisting drought stress, and can assist the rice in resisting the drought stress by regulating the ways of superoxide dismutase, peroxidase, heat shock protein 70 (HSP 70) and NADPH oxidase. These studies are useful in elucidating the antioxidant substance basis of endophytic fungi in application, and lay the foundation for developing novel antioxidants, defining antioxidant stress metabolic pathways of plants, and improving the resistance of higher plants to oxidative stress.
Description
Technical Field
The invention relates to a separation and purification method of secondary metabolites of endogenous aspergillus fumigatus and application thereof, wherein Ultraviolet (UV), Mass Spectrum (MS) and Nuclear Magnetic Resonance (NMR) tests are carried out to determine a structure, experiments prove that the structure can reduce Pro and MDA contents of drought rice, and the rice is assisted to resist drought stress by regulating SOD, POD, HSP70 and NADPH oxidase.
Background
The native asian tropical rice (Oryza sativa) is gradually spread to all over the world after being widely planted in china. Nearly half of the world population all eat rice, so the planting and production of rice have important influence on the national civilian life.
Aspergillus fumigatus belongs to fungi of fungi imperfecti, is mainly present in grains, polluted food, soil and mould rotten objects or is deposited on seeds, can cause pulmonary aspergillosis and other diseases of people, livestock and poultry, and can infect boll, apple and the like to cause fruit rot. The strain can produce various secondary metabolites such as fumigalacins (fumigalacines), fumagilins (fumiremorgins), fumagilins (fumigillins), fumagilins (fumixins), penicillium verruculogen (verruculogen) and the like, but the growth promotion effect of the fumagilins on plants is rarely reported at home and abroad at present.
NFA is useful as a novel antibiotic, is selective against Mycobacterium tuberculosis, also has inhibitory activity against platelet aggregation in rabbits stimulated by arachidonic acid and collagen, has an IC50 value of 2.25, 6.00tag/mL in order, reduces the mortality induced by arachidonic acid in mice by a single intraperitoneal dose of the substance, is biosynthesized from tyrosine and glycine produced by the tetrahydrofolate metabolic pathway, and exhibits antagonism against E.coli while exhibiting moderate cytotoxicity against Du145, A-549 and HeLa cell lines.
The invention separates and purifies a aspergillus fumigatus SG-17 product to obtain a substance NFA with antioxidant activity, which can assist rice to resist drought stress by regulating the contents of Pro, MDA, SOD, POD, HSP70 and NADPH oxidase. Can be used for developing novel antioxidants and planting and producing rice.
Disclosure of Invention
One of the purposes of the invention is to provide a separation and purification process and preparation conditions of NFA with antioxidant activity, and a molecular structure thereof, which serve to develop a novel antioxidant.
An antioxidant product of Aspergillus fumigatus is (Z) -N- (4-hydroxystyryl) formamide. In the process of separation and identification, the NFA of the invention has gradual change phenomenon after 1 day at room temperature, and generates isomer (E) -N- (4-hydroxystyryl) formamide (ENFA). When NFA is isolated by Thin Layer Chromatography (TLC), ENFA appears below the NFA. The Aspergillus fumigatus antioxidant product is characterized by being prepared by using Aspergillus fumigatus SG 17; the preservation unit: china center for type culture Collection; the preservation address is China, Wuhan and Wuhan university; the preservation date is as follows: 5 months and 11 days 2015; the preservation number is CCTCC NO: m2015286.
The invention provides a modifier for improving drought stress resistance of rice, which is (Z) -N- (4-hydroxystyryl) formamide.
The biological modifier for rice contains an antioxidant product (Z) -N- (4-hydroxystyryl) formamide, and the using concentration of the antioxidant is as follows: 0.05 to 0.2mg/mL, and more preferably 0.1 mg/mL.
The separation method of the antioxidant product (Z) -N- (4-hydroxystyryl) formamide comprises the following steps:
(1) strain activation and fermentation culture: inoculating endophytic fungus aspergillus fumigatus SG-17 on a PDA solid culture medium, when a phenotype single colony grows, selecting spores and putting the spores in an SDA liquid fermentation culture medium for culture for 14d to obtain endophytic fungus fermentation bacteria suspension;
(2) preparing a fermentation liquor ester phase primary extract: vacuum filtering and separating endophytic fungi fermentation bacteria suspension to obtain thalli and filtrate; extracting the filtrate with ethyl acetate to obtain a fermentation liquor ethyl acetate phase, rotatably steaming the fermentation liquor ethyl acetate phase to be viscous, and collecting the fermentation liquor ethyl acetate phase as a primary extract of the fermentation liquor ester phase;
(3) TLC separation fermentation liquor ester phase primary extract: mixing petroleum ether and acetone in a volume ratio of 1: 1-2, separating the primary extract of the ester phase of the fermentation liquor to obtain a separated sample; when the fragments separated from the separated sample are multiple fragments, the antioxidant activity of the fragments is measured by adopting a T-AOC method and a DPPH free radical scavenging method, and high performance liquid chromatography analysis is carried out when the antioxidant activity is high.
(4) High performance liquid chromatography analysis: and (3) completely dissolving the separated sample with methanol, filtering with a microporous filter, and performing semi-preparative HPLC separation and purification on the filtrate to obtain the compound NFA.
Chromatographic separation and purification conditions: the analytical column is Cosmosil MS-II RP-C18Column 5 μm, 250 × 10mm analytical, mobile phase: acetonitrile: 60 parts of water: 40(V/V), flow rate: 3.0mL/min, ultraviolet detection wavelength: 254 nm; when NFA is separated by semi-preparative HPLC, the ENFA retention time is about 9.58-10.00min, and the NFA retention time is about 10.78-11.30 min.
Stress tests show that the NFA can obviously reduce the Pro and MDA contents of the drought rice and can effectively assist the rice to resist drought stress.
The NFA has the capability of regulating SOD, POD, HSP70 and NADPH oxidase of rice, can increase the contents of SOD, POD, HSP70 and NADPH oxidase of the rice in a drought state, has obvious effect on improving the drought capability of the rice, and lays a foundation for determining the oxidative stress resisting metabolic pathway of plants and improving the resistance of higher plants to oxidative stress.
Drawings
FIG. 1 structural formulas of NFA and ENFA of the present invention. Wherein the structural formulas of NFA (left) and ENFA (right).
FIG. 2 TLC chart of NFA and ENFA separations of the present invention.
FIG. 3 HPLC profiles of NFA and ENFA of the present invention.
FIG. 4 is a graph showing the effect of NFA of the present invention on drought-stressed rice.
FIG. 5 Effect of NFAs of the invention on rice drought physiology.
FIG. 6 Effect of NFAs of the present invention on oxidative metabolism of rice under drought stress.
Detailed Description
The first embodiment is as follows: the separation and purification process and preparation conditions of the antioxidant active substance NFA, and the molecular structure thereof.
Strain activation and fermentation culture: inoculating endophytic fungus Aspergillus fumigatus SG-17 stored in a refrigerator on a PDA solid culture medium, culturing for 5d in an environment at 28 ℃, picking spores by using a sterile inoculating needle, placing the spores in 150mL SDA liquid fermentation medium (500mL conical flask) when a phenotype single colony grows, and culturing for 14d at 28 ℃ and 120 rmp.
Preparing a crude extract of an ester phase of a fermentation liquid: and filtering and separating the endophytic fungi fermentation bacteria suspension by using a vacuum filtration device to obtain thalli and filtrate. Extracting the filtrate with equal amount of ethyl acetate for 3 times, and mixing the extractive solutions to obtain fermentation liquid ethyl acetate phase. And (3) rotatably steaming the fermentation liquor ethyl acetate phase at 42 ℃ and 0.1MPa to be viscous, and collecting for later use to obtain a primary extract of the fermentation liquor ester phase of the endophytic fungi.
TLC separation SG-17 fermentation liquor ester phase primary extract: separation conditions are as follows: petroleum ether: acetone ═ 1: 1 (volume ratio), and 4 separated fragments (Fr.1-Fr.4) are obtained after the primary extract of the ester phase of SG-17 fermentation liquor is separated. The antioxidant activity of Fr.1-Fr.4 is measured by T-AOC method and DPPH free radical scavenging method, and Fr.2 has the highest antioxidant capacity, so the fragment is taken to be prepared by semi-preparative HPLC. (results are shown below)
High Performance Liquid Chromatography (HPLC) analysis fr.2: taking a small amount of Fr.2 sample, completely dissolving with a small amount of methanol, filtering with a 0.22 μm organic microporous filter, and separating and purifying the filtrate by semi-preparative HPLC to obtain compound NFA. Chromatographic separation and purification conditions: the analytical column is Cosmosil MS-II RP-C18Column (5 μm, 250 × 10mm analytical), mobile phase: acetonitrile: 60 parts of water: 40(V/V), flow rate: 3.0mL/min, ultraviolet detection wavelength: 254 nm. When NFA is separated by semi-preparative HPLC, the ENFA retention time is about 9.58-10.00min, and the NFA retention time is about 10.78-11.30 min.
Conversion of NFA to ENFA: the NFA shows a gradual change after 1 day at room temperature, producing its isomer ENFA. When TLC is used to separate NFA, ENFA appears below NFA.
And (3) NMR detection: freeze-drying NFA of the monomer compound obtained after HPLC separation and purification by using a freeze-drying instrument, dissolving NFA by using 0.5mL of deuterated DMSO, filling the solution into a nuclear magnetic tube, and measuring the hydrogen spectrum and the carbon spectrum of the NFA by using a Bruker AVANCE 400MHz nuclear magnetic resonance spectrometer, wherein the specific steps are as follows:
example two: NFA (NFA-assisted rice drought stress resistance) concentration determination method
Preparing rice seedlings: soaking the seeds in tap water at 4 deg.C for 4 days to promote their rooting and germination, and replacing water every 12 h. Planting the seedlings in a plastic cup filled with soil after 4 days, watering regularly, and obtaining the Japanese fine rice seedlings after 7 days.
Determination of NFA dosage: according to the report of related documents, 0.1mg of NFA is placed in an EP tube, a little DMSO is added for dissolving, the volume is fixed to 1mL by distilled water, so that a sample is fully dissolved, and an NFA solution with the final concentration of 0.1mg/mL is prepared for standby. Selecting rice seedlings with good growth and consistent size, adding NFA solution into Dr + NFA group, and adding 5 × 10 into Dr + Pro group-5mol/L proline solution, 0.1mg/mL Vc solution was added to the Dr + Vc group, tap water was added to the normal group, no solution was added to the Dr group, and each group was repeated 3 times. And (3) culturing the seedlings at room temperature to naturally reduce the water content in soil of each seedling, observing the growth state of each group of rice seedlings, and taking a picture after 20 days to record.
The experimental results are shown in the figure: (A) normal group rice seedlings, (B) Dr group rice seedlings, (C) Dr + NFA group rice seedlings, (D) Dr + Pro group rice seedlings, and (E) Dr + Vc group rice seedlings. The results show that the NFA with the concentration of 0.1mg/mL can assist the rice to resist drought stress, the effect is equivalent to that of Pro and Vc, the phenotype difference of the rice is obvious, the dosage is small, economy is saved, and therefore the concentration is selected for carrying out later-stage experiments (figure 4).
Example three: effect of NFA on Rice drought physiology
Drought stress test: putting 0.1mg NFA into an EP tube, adding a little DMSO for dissolving, and fixing the volume to 1mL by using distilled water to fully dissolve the sample to prepare an NFA solution with the final concentration of 0.1mg/mL for later use. Selecting rice seedlings with good growth and consistent size, adding NFA solution into-Dr + NFA and Dr + NFA groups, and adding 5 × 10 to Dr + Pro group-5mol/L proline solution, add 0.1mg/mL Vc solution to Dr + Vc group, add tap water solution to-Dr group, Dr group does not add any solution, each group is repeated 3 times. One day before drought stress 50mL of the corresponding solution was added to-Dr + NFA, Dr + Pro, Dr + Vc, -Dr group seedlings and Dr group seedlings50mL of tap water was added and only-Dr + NFA, -Dr groups of seedlings were supplemented with 50mL of tap water during the stress period. And (3) culturing the seedlings at room temperature to naturally reduce the water in the soil of each seedling, and determining the Pro and MDA contents of the rice after drought stress for 20 days to determine the influence of NFA on the drought physiology of the rice.
The experimental results show that: dr + NFA is significantly reduced from Dr Pro. As the Pro content of the plant is increased when the plant is subjected to stress injury, the NFA can effectively assist the rice to resist drought stress. Pro content of Dr + NFA is obviously reduced compared with Dr + Pro, and the effect of NFA in assisting rice in resisting drought stress is better than that of Pro. Pro content of Dr + NFA is obviously reduced compared with Dr + Vc, which indicates that the NFA is related to the resistance of rice to drought stress and oxidation resistance.
Dr + NFA is significantly lower than Dr in MDA content. As the MDA content of the plant is increased when the plant is subjected to stress injury, the NFA can effectively assist the rice to resist drought stress. Dr + NFA contained slightly more MDA than Dr + Pro and Dr + NFA contained slightly more MDA than Dr + Vc, indicating that NFA assists rice in resisting drought stress and is related to oxidation resistance (FIG. 5).
Example four: NFA regulates oxidative homeostasis of rice to combat drought stress
Drought stress test: after NFA is extracted, a solution of 0.1mg/mL is prepared, the solution is added into rice seedlings, the contents of SOD, POD, HSP70 and NADPH oxidase of rice are measured at the 5 th, 10 th and 20 th days of drought stress respectively, and the relation between the NFA and the oxidation steady state under the drought stress is revealed.
SOD content: SOD is a substance which can cause superoxide to generate O in the plant body through the disproportionation reaction2And H2O2The antioxidant of (1). It can resist and block the damage of oxygen free radical to cell, repair damaged cell and restore the damage of free radical to cell. Therefore, the determination of SOD can reflect the change of plant metabolism and stress resistance in a certain time. Enzyme-linked immunoassay using plant superoxide dismutase (SOD) from Jiangsu Baolai Biotechnology Ltd
And (3) determining by using a kit, wherein the kit is used for measuring the SOD level of plants by using a double-antibody sandwich method.
POD content: POD is universally produced in plantsAnd the enzyme activity is higher. H in redox reactions2O2Catalyzed by this enzyme, H in the course of oxidation of other substances2O2By being reduced to H2O removing H in body2O2Thus, POD is one of the protective enzymes in plant cells. In addition, processes such as oxidation, respiration, photosynthesis, and formation of lignin of auxin are involved in POD. The POD measurement can reflect the change of the metabolism and the stress resistance of the plants in a certain time. The determination is carried out by utilizing a plant peroxidase determination kit of Jiangsu Baolai Biotechnology limited company, and the kit is used for measuring the content of plant POD by utilizing a double-antibody sandwich method.
HSP70 content: HSP70 is an important stress protein produced by plants under the stress of high temperature, cold, salt, drought, diseases, mechanical wounds and the like, or some chemical substances such as ethanol, methyl jasmonate, heavy metal, arsenic and the like, has the functions of preventing protein polymerization and assisting the refolding of protein without functions, and plays an important role in the aspects of plant growth and development and adversity stress. The determination is carried out by utilizing a plant heat shock protein 70 determination kit of Jiangsu Baolai Biotechnology GmbH, and the kit is used for determining the content of plant HSP70 by utilizing a double-antibody sandwich method.
NADPH oxidase content: NADPH oxidase is a key enzyme for ROS production in the plant organism. When the external factors are induced, the NADPH oxidase can rapidly increase and reduce ROS by utilizing the activation and inactivation of the NADPH oxidase, so that the NADPH oxidase can play a positive role in coping with biotic and abiotic stresses during the growth and development of plants. The determination is carried out by utilizing a plant NADPH oxidase determination kit of Jiangsu Baolai biotechnology limited company, and the kit determines the content of the plant NADPH oxidase by utilizing a double-antibody sandwich method.
And (3) analyzing an experimental result: after drought 15d, the SOD activity was significantly higher in the NFA treated group than in the other groups (P <0.01) (fig. 6A). The change in POD enzyme activity had a similar trend within 10 days (FIG. 6B). SOD and POD are important ROS scavengers for rice under drought stress, so that NFA can help rice to resist drought stress by regulating oxidation stability through antioxidase. Once plants suffer from drought, NADPH oxidase is induced to produce some amount of ROS. The rice after 5 days of drought had a much higher NADPH oxidase content after treatment with NFA (fig. 6C). The result shows that NFA can up-regulate NADPH oxidase to generate ROS in the early stage of drought, and further participate in stress-resistant metabolism. Also in the early stage of drought, NFA can induce the expression of HSP70, which is widely involved in ROS metabolism (fig. 6D), further suggesting that NFA can improve drought resistance of rice by regulating oxidative homeostasis.
Claims (9)
- Application of (Z) -N- (4-hydroxystyryl) formamide in preparing a modifier for improving drought stress resistance of rice.
- 2. Use according to claim 1, characterized in that (Z) -N- (4-hydroxystyryl) formamide is an antioxidant product of aspergillus fumigatus.
- 3. The use according to claim 2, wherein the Aspergillus fumigatus has the deposited name Aspergillus fumigatusAspergillus fumigatusSG 17; the preservation unit: china center for type culture Collection; the preservation address is China, Wuhan and Wuhan university; the preservation date is as follows: 5 months and 11 days 2015; the preservation number is CCTCC NO: m2015286.
- 4. Use according to claim 1, characterized in that (Z) -N- (4-hydroxystyryl) formamide is used in a concentration of 0.05-0.2 mg/mL.
- 5. The use according to claim 4, characterized in that (Z) -N- (4-hydroxystyryl) formamide is used in a concentration of 0.1 mg/mL.
- 6. Use according to any one of claims 1 to 5, characterized in that the process for the isolation of (Z) -N- (4-hydroxystyryl) formamide comprises the following steps:(1) strain activation and fermentation culture: inoculating endophytic fungus aspergillus fumigatus SG17 on a PDA solid culture medium, when a phenotype single colony grows, selecting spores and putting the spores in an SDA liquid fermentation culture medium for culture to obtain endophytic fungus zymophyte suspension;(2) preparing a fermentation liquor ester phase primary extract: vacuum filtering and separating endophytic fungi fermentation bacteria suspension to obtain thalli and filtrate; extracting the filtrate with ethyl acetate to obtain a fermentation liquor ethyl acetate phase, rotatably steaming the fermentation liquor ethyl acetate phase to be viscous, and collecting the fermentation liquor ethyl acetate phase as a primary extract of the fermentation liquor ester phase;(3) TLC separation fermentation liquor ester phase primary extract: separating the primary extract of the ester phase of the fermentation liquor by using a mixed solution of petroleum ether and acetone to obtain a separated sample;(4) high performance liquid chromatography analysis: completely dissolving a separation sample with methanol, filtering with a microporous filter, and performing semi-preparative HPLC separation and purification on the filtrate to obtain a compound NFA, namely (Z) -N- (4-hydroxystyryl) formamide;the collection name of the endophytic fungus aspergillus fumigatus isAspergillus fumigatusSG 17; the preservation unit: china center for type culture Collection; the preservation address is China, Wuhan and Wuhan university; the preservation date is as follows: 5 months and 11 days 2015; the preservation number is CCTCC NO: m2015286.
- 7. The use according to claim 6, wherein, when the fragments separated from the sample in the step (3) are in a plurality of fractions, the antioxidant activity of the plurality of fractions is measured by a T-AOC method and a DPPH free radical scavenging method, and high performance liquid chromatography is performed when the antioxidant activity is high.
- 8. The use according to claim 6, wherein the volume ratio of petroleum ether to acetone is 1: 1-3.
- 9. The use according to claim 6, characterized in that the chromatographic separation and purification conditions are: the analytical column is Cosmosil MS-IIRP-C18Column 5 μm, 250 × 10mm analytical, mobile phase: acetonitrile: water = 60: 40V/V, flow rate: 3.0mL/min, ultraviolet detection wavelength: 254 nm; the retention time of ENFA, i.e. (E) -N- (4-hydroxystyryl) formamide, and the retention time of NFA, i.e. (Z) -N- (4-hydroxystyryl) formamide, are 9.58-10.00min in semi-preparative HPLCIs 10.78-11.30 min.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59175891A (en) * | 1983-03-28 | 1984-10-04 | Fujisawa Pharmaceut Co Ltd | 4-(2-formylaminovinyl)phenol, its salt and its preparation |
| WO2010094666A3 (en) * | 2009-02-17 | 2010-12-23 | Bayer Cropscience Ag | Fungicidal n-(phenylcycloalkyl)carboxamide, n-(benzylcycloalkyl)carboxamide and thiocarboxamide derivatives |
| CN105219651A (en) * | 2015-08-17 | 2016-01-06 | 三峡大学 | A kind ofly improve Rice Resistance arid, interior raw Aspergillus fumigatus that salt damage coerces ability and application thereof |
-
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- 2017-11-27 CN CN201711208611.5A patent/CN107935880B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59175891A (en) * | 1983-03-28 | 1984-10-04 | Fujisawa Pharmaceut Co Ltd | 4-(2-formylaminovinyl)phenol, its salt and its preparation |
| WO2010094666A3 (en) * | 2009-02-17 | 2010-12-23 | Bayer Cropscience Ag | Fungicidal n-(phenylcycloalkyl)carboxamide, n-(benzylcycloalkyl)carboxamide and thiocarboxamide derivatives |
| CN105219651A (en) * | 2015-08-17 | 2016-01-06 | 三峡大学 | A kind ofly improve Rice Resistance arid, interior raw Aspergillus fumigatus that salt damage coerces ability and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| STUDIES ON WF-5239, A NEW POTENT PLATELET AGGREGATION INHIBITOR;KAZUYOSHI UMEHARA等;《THE JOURNAL OF ANTIBIOTICS》;19840531;第XXXVII卷(第5期);第469-474页 * |
| 曲鹏等.黄河三角洲耐盐真菌Penicillium chrysogenum HK14-01的次生代谢产物.《微生物学报》.2012,第52卷(第9期),第1103-1112页. * |
| 黄河三角洲耐盐真菌Penicillium chrysogenum HK14-01的次生代谢产物;曲鹏等;《微生物学报》;20120904;第52卷(第9期);第1103-1112页 * |
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