CN105567726A - Construction method for single knock-out mutant bacterial strain producing deoxynivalenol in high-yield mode - Google Patents
Construction method for single knock-out mutant bacterial strain producing deoxynivalenol in high-yield mode Download PDFInfo
- Publication number
- CN105567726A CN105567726A CN201610041168.6A CN201610041168A CN105567726A CN 105567726 A CN105567726 A CN 105567726A CN 201610041168 A CN201610041168 A CN 201610041168A CN 105567726 A CN105567726 A CN 105567726A
- Authority
- CN
- China
- Prior art keywords
- gene
- deoxynivalenol
- fusarium graminearum
- seqidno
- bacterial strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/04—Phosphoric diester hydrolases (3.1.4)
- C12Y301/04001—Phosphodiesterase I (3.1.4.1)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A construction method for a single knock-out mutant bacterial strain producing deoxynivalenol in a high-yield mode includes the following steps of firstly, knocking a negative regulatory factor high-affinity phosphodiesterase gene FgPDE2 generated in the fusarium graminearumout deoxynivalenol synthesis process out of a genome of fusarium graminearum PH-1 through a split-PCR method, and obtaining the single knock-out mutant, wherein the gene FgPDE2 and upstream and downstream sequences are shown in SEQ ID NO:1; secondly, PEG-mediated protoplast is converted into fusarium graminearum, knocking out converters through resistance screening and marker screening, and verifying that the target gene knocking out the converters is replaced with a neomycin resistance gene through primers 5F and 6R with the genome of fusarium graminearum serving as the reference. The toxicity production amount of the single knock-out mutant bacterial strain is greatly increased; the toxicity production amount of a double knock-out mutant bacterial strain is higher; the gene modification mutant loses the capacity for infecting wheatears, and negative effects caused by possible diffusion of the double knock-out mutant bacterial strain are greatly reduced.
Description
Technical field
The construction process knocking out mutants which had of a kind of high yield deoxynivalenol of the present invention, is specifically related to a kind of construction process knocking out mutants which had of high yield Fusarium graminearum deoxynivalenol.
Background technology
Fusarium graminearum is the pathogenic fungi causing wheat scab (Fusariumheadblightorscab), not only have a strong impact on crop yield, and the multiple mycotoxins deoxynivalenol (DON) that people and animals are harmful to can be produced, the quality of grain or feed is reduced.Deoxynivalenol (deoxynivalenol, DON) be a kind of sesquiterpenoid, not easily degrade, stability is high, be widespread in nature, be considered to primarily of sickle-like bacteria when lacking nutritive substance, synthesis is also present in head blight morbidity wheat as main natural toxin.DON toxin is one of mycotoxins that pollution rate is the highest, mainly pollutes the cereal crop such as wheat, corn, also pollutes cereal product, as bread, biscuit, wheat dessert etc.; In addition, in the milk, egg of animal, also there is the report that DON is residual.This toxin can suppress eukaryotic cells protein synthesis, destroys the immunity system of humans and animals, can enter food chain, serious threat human and livestock health with food, feed.
China endangers one of the most serious country by DON in the world.Therefore, to the examination of DON toxin in cereals goods and hazard evaluation imperative, need too to carry out on a large scale, on a large scale to the research of DON toxin formation mechenism.In fact, China's inspection and quarantine department attaches great importance to the test and monitoring research of this toxin always, and relevant scientific research is also the focus of head blight researcher always.But, these work are owing to needing to use a large amount of DON toxin standard substance, mostly the price of these toxin standard substance again very expensive (1mg price is about 1000 yuans), and is that, from state's outer vent, these factors seriously limit carrying out of related work thus.Relate to the fields such as food safety, environmental protection, medical treatment due to mycotoxins, demand is extensive.Therefore, the preparation of relevant criterion thing has become the focus of various countries' research in the world.
The work of China in prepared by mycotoxins reference material is started late, current, synthesis mycotoxins standard substance mainly utilize the method for chemosynthesis, obtained by preparation technologies such as strain culturing, fermentation, extraction, purifying, constantly optimize and technique perfect although experienced by, extraction efficiency is significantly improved relative last century, but mycotoxins yield is still quite limited, and purifying is with high costs.Therefore, mycotoxins extracting and preparing technique process optimization space is more and more less, and researchist has started to start with seeking breakthrough from screening high yield toxic bacterial strain.The core material that bacterial strain is prepared as toxin, its Toxin producing C determines production efficiency, purity and the preparation cost of preparing material.At present, Fusarium graminearum is the main toxigenic bacterium kind of DON, and China researchist is mainly through to Different sources, and the Toxigenic fungi of different sources carries out being separated and the mensuration of toxin, and then screening is applicable to industrial toxigenic bacterium strain.But the Fusarium graminearum Toxin producing C difference that occurring in nature obtains is little, the bacterial strain being difficult to filter out high yield poison amount is applied to scale production.
Now, genetic modification technology is growing, and in a lot of species, had genetic modification to obtain the trial of breeding.Therefore, develop a kind of construction process singly knocking out mutants which had of high yield deoxynivalenol, can large-scale application in industrial production, be the task of top priority of microorganism resource research.
Summary of the invention
Technical problem to be solved by this invention is, provides a kind of construction process singly knocking out mutants which had of high yield deoxynivalenol.
The technical scheme that the present invention solves the employing of its technical problem is that a kind of construction process knocking out mutants which had of high yield deoxynivalenol, comprises the following steps:
(1) gene knockout carrier is built: utilize split-PCR method, negative regulatory factor high-affinity phosphodiesterase gene FgPDE2 in Fusarium graminearum deoxynivalenol building-up process is knocked out from the genome of Fusarium graminearum PH-1, obtains and singly knock out mutant; The gene of FgPDE2 and upstream and downstream sequence are as shown in SEQIDNO:1;
(2) transform, screen, identify: adopt the protoplast transformation of PEG-mediation in Fusarium graminearum (preferred Fusarium graminearum wild-type PH-1 bacterial strain), then adopt Liu Suanyan NEOMYCIN SULPHATE to knock out transformant as resistance screening label screening; Last with the genome of Fusarium graminearum (preferred Fusarium graminearum wild-type PH-1 bacterial strain) for reference, the goal gene utilizing 5F and 6R primer to verify to knock out transformant substitute by neomycin resistance gene.
Further, in step (1), knocked out by the low affinity phosphodiesterase gene FgPDE1 in Fusarium graminearum, acquisition is two knocks out mutant, and the gene of FgPDE1 and upstream and downstream sequence are as shown in SEQIDNO:2 simultaneously.
Further, FgPDE1 the primer sequence PDE1-1F+PDE1-2R is knocked out described in; PDE1-3F+PDE1-4R; YG/F+HY/R; HYG/F+HYG/R, as shown in SEQIDNO:3 ~ 10.
Further, in step (1), described in knock out FgPDE2 the primer sequence PDE2-1F+PDE2-2R, PDE2-3F+PDE2-4R, EN/F+GE/R, GEN/F+GEN/R, as shown in SEQIDNO:11 ~ 18.
Further, in step (2), described 5F and 6R primer is PDE2-5F, and as shown in SEQIDNO:19, PDE2-6F, as shown in SEQIDNO:20; PDE1-5F, as shown in SEQIDNO:21, PDE1-6R, as shown in SEQIDNO:22.
The present invention passes through negative regulatory factor high-affinity phosphodiesterase (phosphodiesterase) gene (FgPDE2 in Fusarium graminearum deoxynivalenol DON building-up process, FGSG_06914), this gene knocks out by the protoplast transformation method utilizing split-PCR and PEG-to mediate from the genome of Fusarium graminearum PH-1, and what acquisition deoxynivalenol resultant quantity significantly improved singly knocks out mutant (genetic modification bacterial strain); And by low affinity phosphodiesterase (phosphodiesterase) gene (FgPDE1 in Fusarium graminearum, FGSG_06633) knock out simultaneously, the two mutant product poison that knock out obtained are measured higher, and the infection ability of this genetic modification mutant loss to wheatear, greatly reduce two adverse consequences knocking out mutants which had possible diffusion and cause.
Test and prove, compared to wild-type, the mono-mutant product poison amount that knocks out of Fgpde2 can improve 67.5 times, and Fgpde1Fgpde2 pair knocks out mutant and then can improve 257 times more than.
The construction process knocking out mutants which had of the high yield deoxynivalenol of the present invention can reach and utilize a small amount of starting material mycelia to synthesize the object of deoxynivalenol in a large number in the industrial production, is a bacterial strain being highly suitable for for the preparation of DON under laboratory condition.
Accompanying drawing explanation
Fig. 1 is the comparing result that in the present invention, genetic modification bacterial strain produces poison amount and wild-type under liquid culture condi.
Fig. 2 is the comparing result that in the present invention, genetic modification bacterial strain produces poison amount and wild-type in rice medium.
Fig. 3 be in the present invention genetic modification bacterial strain on the impact of virulence.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the invention will be further described.
High-affinity phosphodiesterase (phosphodiesterase) gene (FgPDE2 is searched from the genome database (http://www.broadinstitute.org/annotation/genome/fusarium_gramin earum/MultiHome.html) of Fusarium graminearum wild type strain PH-1, FGSG_06914) F and upstream and downstream sequence, the DNA sequence dna of FgPDE2 is as shown in SEQIDNO:1, and highlighted part is the encoder block of gene.Low affinity phosphodiesterase (phosphodiesterase) gene (FgPDE1, FGSG_06633) and upstream and downstream sequence, the DNA sequence dna of FgPDE1 is as shown in SEQIDNO:2, and highlighted part is the encoder block of gene.
Be designed for the primer of amplification FgPDE2 upstream and downstream homologous sequence and neomycin resistance gene, for gene knockout, their sequence is as shown in SEQIDNO:3-10; Be designed for the primer of amplification FgPDE1 upstream and downstream homologous sequence and hygromycin gene, for gene knockout.Their sequence is as shown in SEQIDNO:11-18; Be designed for and detect the positive PCR primer knocking out body, DNA sequence dna is as shown in SEQIDNO:19-22.
Embodiment 1: the structure knocking out mutants which had of high yield deoxynivalenol
(1) with the DNA of Fusarium graminearum wild-type PH-1 bacterial strain for template, utilize primer PDE2-1F+PDE2-2R, PDE2-3F+PDE2-4R amplification gene upstream from start codon U and terminator codon downstream sequence D; With the plasmid PFL2 containing neomycin resistance gene for template, utilize primer GEN/F+GE/R, FN/F+GEN/R amplification neomycin resistance gene sequence G1 and G2; Wherein, pcr amplification reaction system: in 50 microlitre PCR reaction solutions, containing 50 nanogram template DNAs, 10 microlitre 5XPfu damping fluids, 1 microlitre 10 mmole dNTP, 0.5 l primer P1 (10 micromole), 0.5 l primer P2 (10 micromole), 0.4 microlitre FastPfuDNA polysaccharase (5 activity unit/microlitres); PCR reaction conditions: 94 DEG C 2 minutes; 94 DEG C 20 seconds, 55 DEG C 20 seconds, 72 DEG C 40 seconds, 32 circulations; And extend eventually 72 DEG C 5 minutes; After having reacted, electrophoresis on 1% sepharose, takes a picture under gel imaging system medium ultraviolet lamp and carries out cutting to band and reclaim PCR primer; By SpiltPCR method by U and G1 fragment, D with G2 fragment is connected respectively, concentrates above-mentioned connection product by isopropanol precipitating concentration method.
(2) the PH-1 spore in CMC liquid nutrient medium is collected, cultivate 12 hours in YEPD liquid nutrient medium, collecting about 0.5 gram mycelia, 30 DEG C of enzymolysis (2.5% driselase+0.5% lywallzyme) 3 hours, is 4x10 by the protoplastis dilution obtained
7individual/milliliter, step (1) is recycled product conversion and enters in protoplastis by the method mediated by PEG, and recovers to protoplastis with TB3 liquid nutrient medium.
(3) after protoplastis recovery growth 16h, proceed in solid medium, screen with the Liu Suanyan NEOMYCIN SULPHATE of 300 mg/litre final concentrations, after son to be transformed grows solid medium, picking also proceeds to new substratum, extracts transformant DNA and utilize 5F and 6R primer to detect further to confirm transformant.
On the basis of the genetic modification bacterial strain obtained, use the same method and knock out FgPDF1 further, adopt Totomycin to mark as resistance screening, acquisition is two knocks out mutant.
Embodiment 2: the toxin resultant quantity evaluating genetic modification mutant
1, liquid culture produces malicious method: the spore obtaining wild-type and mutant, joins liquid toxin producing medium (liquid toxin producing medium (1 liter): 30 grams of sucrose, 1 gram of SODIUMNITRATE, 1 gram of primary ammonium phosphate, 0.5 gram of magnesium sulfate heptahydrate, 0.5 gram of Repone K, 10 milligrams of iron vitriol, 0.03% plant gel and 200 microlitres trace element mixed solution (micro-mixed solutions (100 milliliters): 5g citric acid, 5 grams of Zinc Sulphate Heptahydrates, 0.25 gram of cupric sulfate pentahydrate, 50 milligrams of manganese sulfate monohydrates, 50 milligrams of boric acid, 50 milligrams of Sodium Molybdate Dihydrates.), pH value sodium hydroxide is adjusted to 6.5.) to final concentration 10
4individual spore/milliliter, dark quiescent culture 7 days, measures the toxin in substratum.The ELISA kit that toxin determination utilizes Beacon company to produce, concrete operations are: draw substratum, centrifugal segregation mycelia, obtain sample; Add enzyme, sample, antibody successively in the reaction cup be equipped with in test kit, mix, standing and reacting 10 minutes, abandons reaction solution, with the scavenging solution in test kit, washes reaction cup 5 times, adds substrate reactions 5 minutes, use stop buffer termination reaction; OD is surveyed in microplate reader
450numerical value also contrasts with the typical curve that standard specimen is done, and calculates concrete numerical value.
Experimental result is shown in Fig. 1, and as shown in Figure 1, compared to wild-type, the mono-mutant product poison amount that knocks out of Fgpde2 improves 67.5 times, and Fgpde1Fgpde2 pair knocks out mutant and then improve 257 times more than.
2, malicious method is produced in rice induction: be placed in 50 milliliters of triangular flasks by 6 grams of rice, adds 2 ml sterile waters and to soak after 2 hours 121 DEG C of sterilizings 1 hour; The bacterial strain activated in inoculation PDA substratum, connects the bacterium cake of 35 mm dias, is placed in 25 degree and cultivates 21 days in each bottle; Freeze drying example spends the night, and takes 4 grams of samples measure for DON with liquid nitrogen after fully grinding, and be placed in 50 milliliters of centrifuge tubes and add the acetonitrile/water (84: 16) of 16 milliliters, on shaking table, level shakes 1 hour; Filtered sample, (3 milliliters of plastics pillars, fill 500 milligrams of C18/ neutral alumina=1: 3), collect filtrate and are transferred to centrifuge tube, 50 degree of dried overnight to get filtrate 3 milliliters mistake post; Add TMS (TMSI/TMCS=100/1) silylating reagent of 100 microlitres, shake 10 minutes; Add the chromatographic grade octane-iso of 800 microlitres, slightly turn upside down and make abundant mixing, add ultrapure water 800 microlitre, slightly turn upside down and make sample become clarification and layering; Leave standstill 10 minutes, 700 microlitre supernatants are all transferred in GC loading bottle, gas chromatograph-mass spectrometer measures content of toxins.
Experimental result is shown in Fig. 2, and as shown in Figure 2, compared to wild-type, the mono-mutant product poison amount that knocks out of Fgpde2 improves 9.8 times, and Fgpde1Fgpde2 pair knocks out mutant and then improve 12.4 times.
In addition, applicant also carries out identifying to the virulence of genetic modification mutant and analyzes, and sees Fig. 3.As shown in Figure 3, Fgpde2 is mono-knocks out mutant virulence with wild-type difference not quite, but two virulence knocking out mutant almost completely loses.
To sum up, although this bacterial strain can volume production be malicious greatly, wheatear cannot be infected.Therefore, this bacterial strain is harmless to farm crop, greatly reduces the risk of instability because possible diffusion brings, and is a bacterial strain being highly suitable for for the preparation of DON under laboratory condition.
Claims (5)
1. the construction process knocking out mutants which had of high yield deoxynivalenol, is characterized in that, comprise the following steps:
(1) gene knockout carrier is built: utilize split-PCR method, negative regulatory factor high-affinity phosphodiesterase gene FgPDE2 in Fusarium graminearum deoxynivalenol building-up process is knocked out from the genome of Fusarium graminearum PH-1, obtains and singly knock out mutant; The gene of FgPDE2 and upstream and downstream sequence are as shown in SEQIDNO:1;
(2) transform, screen, identify: adopt the protoplast transformation of PEG-mediation in Fusarium graminearum, then adopt resistance screening label screening to knock out transformant; Last with the genome of Fusarium graminearum for reference, the goal gene utilizing 5F and 6R primer to verify to knock out transformant substitute by neomycin resistance gene.
2. the construction process knocking out mutants which had of high yield deoxynivalenol according to claim 1, it is characterized in that, in step (1), low affinity phosphodiesterase gene FgPDE1 in Fusarium graminearum is knocked out simultaneously, acquisition is two knocks out mutant, and the gene of FgPDE1 and upstream and downstream sequence are as shown in SEQIDNO:2.
3. the construction process knocking out mutants which had of high yield deoxynivalenol according to claim 2, is characterized in that, described in knock out FgPDE1 the primer sequence PDE1-1F+PDE1-2R; PDE1-3F+PDE1-4R; YG/F+HY/R; HYG/F+HYG/R, as shown in SEQIDNO:3 ~ 10.
4. the construction process knocking out mutants which had of high yield deoxynivalenol according to claim 1, it is characterized in that, in step (1), describedly knock out FgPDE2 the primer sequence PDE2-1F+PDE2-2R, PDE2-3F+PDE2-4R, EN/F+GE/R, GEN/F+GEN/R, as shown in SEQIDNO:11 ~ 18.
5. according to the construction process knocking out mutants which had of the high yield deoxynivalenol one of Claims 1 to 4 Suo Shu, it is characterized in that, in step (2), described Fusarium graminearum is Fusarium graminearum wild-type PH-1 bacterial strain; Described 5F and 6R primer is PDE2-5F, and as shown in SEQIDNO:19, PDE2-6F, as shown in SEQIDNO:20; PDE1-5F, as shown in SEQIDNO:21, PDE1-6R, as shown in SEQIDNO:22; Described resistance screening mark knocks out for FgPDE2, selects Liu Suanyan NEOMYCIN SULPHATE; FgPDE1 is knocked out, selects Totomycin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610041168.6A CN105567726B (en) | 2016-01-21 | 2016-01-21 | A kind of construction method of the knockout mutations body bacterial strain of high yield deoxynivalenol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610041168.6A CN105567726B (en) | 2016-01-21 | 2016-01-21 | A kind of construction method of the knockout mutations body bacterial strain of high yield deoxynivalenol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105567726A true CN105567726A (en) | 2016-05-11 |
| CN105567726B CN105567726B (en) | 2019-10-22 |
Family
ID=55878350
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610041168.6A Expired - Fee Related CN105567726B (en) | 2016-01-21 | 2016-01-21 | A kind of construction method of the knockout mutations body bacterial strain of high yield deoxynivalenol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105567726B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109777744A (en) * | 2019-01-22 | 2019-05-21 | 浙江大学 | A kind of method for the compound that quick screening inhibits gibberellic hypha DON toxin to synthesize |
| CN110616155A (en) * | 2019-10-15 | 2019-12-27 | 江苏省农业科学院 | Wild strain for high yield of nivalenol and construction method of mutant thereof |
-
2016
- 2016-01-21 CN CN201610041168.6A patent/CN105567726B/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| CHRISTINA A. CUOMO等: "The Fusarium graminearum Genome Reveals a Link Between Localized Polymorphism and Pathogen Specialization", 《SCIENCE》 * |
| RUI HOU等: "The AreA transcription factor mediates the regulation of deoxynivalenol (DON) synthesis by ammonium and cyclic adenosine monophosphate (cAMP) signalling in Fusarium graminearum", 《MOLECULAR PLANT PATHOLOGY》 * |
| 常玉梅 侯占铭: "禾谷镰刀菌中FgPDE1基因的敲除及其功能的研究", 《中国生物工程杂志》 * |
| 张岳平: "镰刀菌真菌毒素产生与调控机制研究进展", 《生命科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109777744A (en) * | 2019-01-22 | 2019-05-21 | 浙江大学 | A kind of method for the compound that quick screening inhibits gibberellic hypha DON toxin to synthesize |
| CN110616155A (en) * | 2019-10-15 | 2019-12-27 | 江苏省农业科学院 | Wild strain for high yield of nivalenol and construction method of mutant thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105567726B (en) | 2019-10-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106591156B (en) | One plant of epicoccum nigrum FXZ2 and its application | |
| CN106916765B (en) | A method of heavy metal in waste water zinc is adsorbed using penicillium janthinellum | |
| CN109825444B (en) | Strain for improving food safety of red yeast rice yellow wine and brewing method thereof | |
| CN108841733A (en) | The one plant of production main part of Songgangmeisu --- bacterial strain and method of griseofulvin | |
| CN109943488A (en) | A kind of ganoderma lucidum polysaccharide superior strain RWHBW-1 and its application | |
| CN107502566B (en) | Lysine bacillus and application thereof in degradation of zearalenone | |
| CN105567726A (en) | Construction method for single knock-out mutant bacterial strain producing deoxynivalenol in high-yield mode | |
| CN102094080A (en) | Quick molecular detection method for simultaneously detecting three kinds of fusarium toxins and application thereof | |
| CN113607959B (en) | Rapid detection kit for aflatoxin toxigenic bacteria toxigenic indicator molecule immunity and application thereof | |
| CN102925361A (en) | AFB1 degrading bacterium and degrading enzyme | |
| CN110295211A (en) | A kind of preparation method and application of bacterium selenium-enriched protein | |
| CN106676044B (en) | One plant of Rhodopseudomonas palustris and its application | |
| CN109385488A (en) | Multiple PCR primer, method and the kit of Chinese medicine pollution three classes Toxigenic fungi are detected simultaneously | |
| CN1312475C (en) | Molecular detection method for fusarium toxin | |
| CN116536212A (en) | Serratia AWH-NS6 and application thereof in dissolving phosphorus and promoting plant growth | |
| CN108130281B (en) | Aspergillus terreus DDT98801 and its screening technique and the application in degradation of dichloro-diphenyl-trichloroethane | |
| CN102443589A (en) | Molecular identification method of trichothecene type-B toxins of fusarium | |
| CN112725193B (en) | Trichoderma tomentosum and application thereof | |
| CN104894042B (en) | The escherichia coli of one plant of detection arsenic | |
| CN116144522A (en) | Paenibacillus pumilus and application thereof in preventing and treating white rot of grape | |
| CN105483175A (en) | Inducer and method for synthesizing fusarium graminearum deoxynivalenol | |
| CN110343620B (en) | Plant rhizosphere fungus F9 capable of adsorbing cadmium ions and application thereof | |
| CN106497794B (en) | One plant of naked born of the same parents' shell of structure nest for producing echinocandin B and its application | |
| CN106011220B (en) | The method for quickly separating and detecting and used medium of sickle-like bacteria in a kind of sample | |
| CN109182154B (en) | Pabuvina rhodotorula strain capable of producing protease at high yield |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20191022 Termination date: 20210121 |