CN107501086A - One kind comes from 16 carbon chain fatty acid class antagonistic substances and its application caused by bacillus amyloliquefaciens SQR9 - Google Patents
One kind comes from 16 carbon chain fatty acid class antagonistic substances and its application caused by bacillus amyloliquefaciens SQR9 Download PDFInfo
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Abstract
The invention discloses one kind to come from 16 carbon chain fatty acid class antagonistic substances and its application caused by bacillus amyloliquefaciens SQR9.The 16 carbon chain fatty acid class antagonistic substance molecular formula is C17H34O3, there is has a broad antifungal spectrum, have the advantages of good growth inhibitory activity to plant pathogenic fungi and pathogenetic bacteria, it is notable to the growth inhibitory effect of disease fungus, bacterium, safety and stability, it is with a wide range of applications；It can be applied in plant pathogenic fungi and pathogenetic bacteria growth is suppressed.
Description
Technical field
It is specific a kind of from 16 carbochains fat caused by bacillus amyloliquefaciens SQR9 the invention belongs to microorganism field
Acids antagonistic substance and its application.
Technical background
Antibiotic (Antibiotics) is to be lived by microorganism including bacterium, fungi, actinomyces or high animals and plants
During caused by there is antipathogen or other active a kind of secondary metabolites.Abuse of antibiotics makes bacterium gradually produce
Raw drug resistant gene, makes antibiotic medicine effect be deteriorated, or even invalid.Moreover, what bacterial drug resistance can be propagated mutually, make bacterium
Drug resistance complicates, therefore is badly in need of finding the present problems faced of new antibiotic solution.Find the tradition side of new antibiotic
Method is to cultivate the bacterial strain newly separated at different conditions, then fermented product is isolated and purified, and finally identification obtains new antibiosis
Element.But with the development of genomic sequencing technique in the last few years, full-length genome data analysis provides a kind of find and active matter
The approach of matter synthesis related gene cluster.Fatty acid structure is simple, but with various biological activity, one of them is exactly can be with
Suppress the growth of parasite or phytopathogen, there is good application prospect.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) are widely distributed in nature, to people, animal
It is nontoxic, it is free from environmental pollution, there is very high anti-adversity ability.Research shows that bacillus amyloliquefaciens have the suppression of wide spectrum
Fungi and the activity of bacterium, and the speed of growth is fast, stability is strong, is a kind of preferably biocontrol microorganisms resource.Bacillus amyloliquefaciens
Multiple Classes of Antibiotics class material can be secreted, such as lipopeptide antibiotic (surfactin, bacillomycin D, fengycin)；Polyketone
Class antibiotic (macrolactin, difficidin, bacillaene)；Small molecule antibiotic bacilysin etc..
The content of the invention
The purpose of the present invention will solve the problems, such as to be to provide one kind by a kind of 16 carbon of bacillus amyloliquefaciens SQR9 fermentations
Chain fatty acids antagonistic substance.
It is a further object of the present invention to provide the extraction separation method of the antagonistic substance.
It is yet another object of the invention to provide the application of the antagonistic substance.
One kind comes from 16 carbon chain fatty acid class antagonistic substances, the following institute of structure caused by bacillus amyloliquefaciens SQR9
Show:
A kind of method that 16 described carbon chain fatty acid class antagonistic substances are separated in SQR9 from bacillus amyloliquefaciens, bag
Include：The bacillus amyloliquefaciens SQR9 that fermentative preservation numbering is CGMCC NO.5808, after fermentation ends, zymotic fluid is centrifuged, and is received
Collect supernatant, after moisture removal is removed in supernatant drying, extracted with methanol, concentration leaching liquor obtains medicinal extract；Silicagel column on medicinal extract, with chlorine
Imitation-carbinol gradient elution, it is 20 to collect chloroform-methanol volume ratio:1 eluent；By chloroform-methanol 20:1 eluent removes molten
Dissolved after agent with methanol, further with MPLC ODS chromatographic columns methanol-water gradient elution, the first of collected volume fraction 80%
Alcohol-water elution further with rp-hplc method carry out it is deep isolate and purify, in acetonitrile-water 65:35, flow velocity 2mL/min,
Detection wavelength is 210nm, and 16 described carbon chain fatty acid class antagonistic substances are obtained under Rt=12.1.
The preferred Landy of fermentation medium for the bacillus amyloliquefaciens SQR9 that fermentative preservation numbering is CGMCC NO.5808
Culture medium.
Bacillus amyloliquefaciens SQR9 fermentation process preferably includes：SQR9 seed liquors are transferred to by 1% inoculum concentration
In fresh Landy culture medium conical flasks, 37 DEG C, 170rpm, 12h；Bacterium solution after culture is inoculated into 7L small-sized fermentation again
In tank, 5L Landy culture mediums are filled in fermentation tank, 37 DEG C, 200rpm cultivates 12h；Bacterium solution in 7L fermentation tanks is inoculated into
In 150L fermentation tanks, 100L Landy culture mediums are filled in fermentation tank, rotating speed is 30 DEG C, rotating speed 200rpm during beginning, oxygen dissolving value
75, defoamer is polyether compound, when oxygen dissolving value starts to drop to below 30, gradually steps up rotating speed, up to 500rpm, together
Shi Zengjia throughputs, oxygen dissolving value is maintained at more than 10, until OD600=15, is now adjusted to 200rpm, throughput drop by rotating speed
It is as little as minimum, OD is kept, bacterium solution enters stationary phase, starts a large amount of generation secondary metabolites, ferment 24h.
As the preferred of the inventive method：Described medicinal extract adds methanol to make fully to dissolve, and the quality such as lysate adds 100~
200 mesh silica gel make to be sufficiently mixed, and are concentrated under reduced pressure to be drying to obtain and mix sample silica gel；Another 100~200 mesh silicon for weighing 2 times of medicinal extract quality
Glue is placed among glass column as separation silica gel, using chloroform-methanol volume ratio as 1:0；100:1；80:1；60:1；40:1；
20:1 gradient elution.
As the preferred of the inventive method, described method also includes passing through the active component that rp-hplc method is separated to
Gel chromatography is separated except inactive impurity, finally obtains the pure of 16 described carbon chain fatty acid class antagonistic substances
Product.
16 carbon chain fatty acid class antagonistic substance of the present invention is suppressing plant pathogenic fungi and pathogenetic bacteria growth
In application.
The preferred gram-positive bacterium of described pathogenetic bacteria.
16 described carbon chain fatty acid class antagonistic substances further preferably suppress cucumber Fusarium oxysporum, sclerotium bacteria,
Banana Fusarium oxysporum, Phytophthora capsici, potato Rhizoctonia solani Kuhn, Rhizoctonia cerealis and Solanaceae Raul bacterium, Staphylococcus aureus
Application in bacteria growing.
Beneficial effect:
The application obtains the one section of 84kb gene sequence not being reported from bacillus amyloliquefaciens SQR9 full genome
Row, are a gene transferring horizontally island, GI3 are named as in the position of genome according to it.Pass through gene knockout and growth inhibition
Experiment, the activity of validating substance, has groped fermentation of materials optimum medium and condition of culture, and isolates and purifies out a kind of 16 carbon
Chain fatty acid antagonistic substance C17H34O3, identify the molecular structure of material.Empirical tests, the antagonistic substance can suppress cucumber point spore sickle
Knife bacterium, sclerotium bacteria, banana Fusarium oxysporum, Phytophthora capsici, potato Rhizoctonia solani Kuhn, Rhizoctonia cerealis and Solanaceae Raul bacterium
Growth, antimicrobial spectrum are wide.
Biological deposits information
SQR9, Classification And Nomenclature are bacillus amyloliquefaciens Bacillus amyloliquefaciens, and it is micro- to be preserved in China
Biological inoculum preservation administration committee common micro-organisms center, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica, preservation date are on 2 27th, 2012, and preserving number is CGMCC NO.5808.
Embodiment
The invention discloses a kind of novel fatty acid antagonistic substance, those skilled in the art may be referred to present disclosure, fit
When modified parameters are realized.In order that those skilled in the art more fully understands technical scheme, with reference to specific
The present invention is described in further detail for embodiment.
Embodiment 1：Verify growth inhibitory activity of the aliphatic acid of the present invention to homologous strain：
Bacillus amyloliquefaciens SQR9 genomic islands GI3 is a Horizontal transfer genomic islands, positioned at SQR9 genome cores
Nucleotide sequence 657050-738610 positions, the gene that GI3 is included are to be not present in the same position of homologous strain, infer the base
Because the active material of island GI3 synthesis is related to homologous strain.In order to study the function of genomic islands GI3 in SQR9, using double cross
The method for changing homologous recombination has carried out full knockout to genomic islands.Chloramphenicol (Cm) resistance is sieved using the method for fusion DNA vaccine first
The gene of choosing mark merges with the upstream and downstream homology arm of gene to be knocked out, and fusion fragment is transferred in competence SQR9,37
DEG C, 100r min-1Bacteria suspension is coated on LB flat boards (the μ g ml of Cm 5 after recovery culture 8h-1), transformant checking is chosen, is obtained just
True mutant △ GI3.Bacterial strain SQR9 and △ GI3 are connected into Landy culture mediums, and (Landy culture mediums, 1L culture mediums are with method：
1st, glucose 40g, individually sterilizing, 115 DEG C, 30min.2、MgSO4·7H2O 0.5g；KH2PO41g；KCl 0.2g；MnSO4·
H2O 10mg；FeSO4·7H2O 5mg；CuSO4·5H2O 0.2mg；Dusty yeast 1g；ALANINE 2mg；Pidolidone 5g, uses 5M
NaOH modulation pH=6.7 after, 115 DEG C, 30min sterilizing.1st, 2 be sterilized separately after mix) in, 30 DEG C, 200rpm training
Support 72 hours.By 4 DEG C of zymotic fluid, 12000r/min centrifugations 10min removes thalline.Moisture removal is removed in the freeze-drying of supernatant zymotic fluid,
Extracted to obtain crude extract (50ml methanol with methanol：1L zymotic fluid).Crude extract is crossed to 0.22um sterile organic phase filter membrane, gone
Except precipitation, impurity and bacterium, it is stored in -20 DEG C.SQR9 crude extracts are experimental group, and △ GI3 crude extracts are control group.
The type strain FZB42 of bacillus amyloliquefaciens is have selected, bacterium colony face-off experiment is carried out on LB culture mediums.It is preferred
On the flat board of LB culture mediums in uniform sprinkling FZB42 bacterium solution, after natural air drying, Oxford cup is placed in relevant position.Ox
The crude extract of 100ul experimental groups and control group is separately added into the cup of Tianjin.After 4 DEG C stand 8 hours, after being put into 37 DEG C of culture 24h, see
Examine experimental result.
Growth inhibition effects of the wild type SQR9 of the table 1 and mutant △ GI3 to homologous strain FZB42
Experimental result is as shown in table 1, and wild type SQR9 can suppress FZB42 growth, but as mutant △ GI3 lose
After the function of synthesizing this material, FZB42 growth inhibitory effect is wholly absent, illustrating the material of genomic islands GI3 synthesis is
For the homologous strain bacillus amyloliquefaciens class with SQR9.
Embodiment 2：Prepare and isolate and purify aliphatic acid antagonistic substance of the present invention：
Mutant △ GI3 only knock out genomic islands GI3, mutant △ GI3 can not be synthesized ten mentioned in this application again
Six carbon chain fatty acid classes, i.e. C17H34O3, the synthesis without influenceing other active materials in SQR9, so experimental group is poor with control group
Different is C17H34O3Bioactivity.Mutant △ GI3 are wholly absent to FZB42 growth inhibitory effect in embodiment 1, therefore
We can isolate and purify by way of activity tracks to 16 carbon chain fatty acid classes of △ GI3 synthesis, that is, separate
Each step all verifies the growth inhibitory activity to FZB42, and active component further isolates and purifies.
Prepare seed liquor first：Picking SQR9 single bacteriums are fallen within the test tube of Landy culture mediums, 170rpm, 37 DEG C, 12h.Press
Seed liquor is transferred in fresh Landy culture medium conical flasks by 1% inoculum concentration, 37 DEG C, 170rpm, 12h.After cultivating again
Bacterium solution be inoculated into 7L small-sized fermentation tank, in fermentation tank fill 5L Landy culture mediums, 37 DEG C, 200rpm culture 12h.
Bacterium solution in 7L fermentation tanks is inoculated into 150L fermentation tanks, 100L Landy culture mediums are filled in fermentation tank.Rotating speed during beginning
For 30 DEG C, rotating speed 200rpm, oxygen dissolving value 75, defoamer is polyether compound.When oxygen dissolving value starts to drop to below 30, gradually
Rotating speed, up to 500rpm are improved, while increases throughput, oxygen dissolving value is maintained at more than 10, until OD600=15, now
Rotating speed is adjusted to 200rpm, throughput is reduced to minimum, keeps OD, and bacterium solution enters stationary phase, starts a large amount of generation cometabolisms
Product, ferment 24h.
After fermentation ends, zymotic fluid 12000rpm centrifugations remove thalline, collect supernatant, about 90L.After supernatant is dried,
Extracted with methanol, after extraction, rotary evaporation, obtain 123 grams of medicinal extract.Proper amount of methanol is added to make fully to dissolve, lysate adds equivalent silicon
Glue (123 grams, 100~200 mesh) makes to be sufficiently mixed, and is concentrated under reduced pressure to be drying to obtain and mixes sample silica gel.Separately weigh on 2 times of amounts (246 grams)
Used silica gel is stated to be placed among glass column as separation silica gel, with chloroform-methanol (volume ratio 1:0；100:1；80:1；60:
1；40:1；20:1；10:1；0:1) gradient elution, suppression of each gradient eluent checking to bacillus amyloliquefaciens FZB42 is collected
Bacterium activity (method reference implementation example 1), eluent 20:1 have it is more strongly active, by 20:1 eluent removes solvent and obtains activearm
Divide (1.50 grams).The 20 of previous step collection:After 1 component is dissolved with proper amount of methanol, gained medicinal extract is examined respectively using TLC
Know, the colour developing of sulfuric acid ethanol, judge active component medicinal extract ingredient feature for long chain fatty acids；After TLC checkings, by 20:1 component
Further with MPLC ODS chromatographic column methanol：Water (volume ratio 30:70；50:50；60:40；70:30；80:20；90:10；
100:0) gradient elution successively, collecting 80% methanol-water eluent, further to carry out deep separation with rp-hplc method pure
Change, in acetonitrile-water (65:35) it is more significant that activity is obtained under, flow velocity 2mL/min, Detection wavelength 210nm, Rt=12.1
Compound FG-80%- (2).FG-80%- (2) has good bacteriostatic activity to bacillus amyloliquefaciens FZB42, and (method is with reference to real
Apply example 1).Separated further through gel chromatography (85% methanol) except inactive impurity, finally obtain the pure of active component
Product.Mass spectrum and NMR (including two dimensional NMR structure elucidation) tests are carried out to active component, final reactive compound is length
Chain saturated fatty acid class compound, molecular structure are as follows:
The chemical molecules formula that final separation Purification goes out chain saturated fatty acids antagonistic substance is C17H34O3, degree of unsaturation
It is 1, meets long chain fatty acids feature.C17H34O3Simpleness for [M+H-H2O] +=269, [M+H] -=287, [M+Na] +=
309, [M+K] +=325.1H NMR(600MHz,CD3OD) δ 0.86 (3H, d, J=7.2Hz), 0.9 (3H, t, J=7.8Hz),
1.32 (19H, m), 1.41-147 (2H, m), 1.59 (2H, m), 2.27 (2H, t, J=7.2Hz), 3.43 (1H, m)13C NMR
(150MHz,CD3OD)δ13.16(CH3),14.82(CH3),26.99(CH2),27.99(CH2),28.33(CH2),31.14
(2CH2),31.32(2CH2),31.48(CH2),31.62(CH2),31.74(CH2),35.87(CH2),36.27(CH2),42.37
(CH),76.3(-OCH),178.55(-COOH).Calculating finally determines that the molecular formula of the compound is C17H34O3, error is
(9.9ppm)。
Embodiment 3：Verify growth inhibitory activity of the aliphatic acid of the present invention to phytopathogen：
The chain saturated fatty acids solution (100 μ g/ml) that the 7 pathogen strain fungies chosen separate with pathogen strain bacterium checking
Antagonistic activity.Choose 7 pathogen strain bacterium be respectively：
Cucumber Fusarium oxysporum (Fusarium oxysporum.sp.cucumebrium Owen, the separation of this laboratory),
Sclerotium bacteria (Sclerotiniasclerotiorum, the separation of this laboratory),
Banana Fusarium oxysporum (Fusarium oxysporum Schl, the separation of this laboratory),
Phytophthora capsici (Phytophthora capsici, ACCC NO.36278),
Potato Rhizoctonia solani Kuhn (Rhizoctonia solani Ktihn, ACCC NO.36246),
Rhizoctonia cerealis/rhizoctonia cerealis (Rhizoctonia cerealis, ACCC NO.37393),
Corn stem rot Fusarium moniliforme (Fusarium Verticillioides, the separation of this laboratory)
Ralstonia solanacearum (Ralstonia solanacearum, the separation of this laboratory)
Pathogen, is inoculated into PDA culture medium, is positioned over 30 DEG C by fungal pathogens face-off experiment first, treats that mycelia grows
After diameter about 1cm, retell Oxford cup and be placed on culture medium, at mycelia edge 3cm.100ul separation is added in Oxford cup
Chain saturated fatty acids solution (100 μ g/ml).Flat board is placed in 30 DEG C of incubators again, until pathogen mycelia is covered with entirely
Flat board.Bacterial disease opportunistic pathogen is that bacterial disease opportunistic pathogen cuts a gram Raul Salmonella in uniform sprinkling on NA culture medium flat plates, then by ox
Tianjin cup is placed on flat board, adds the chain saturated fatty acids C of 100ul separation in Oxford cup respectively17H34O3Solution (100 μ g/ml), 4
DEG C stand overnight, be then placed into 30 DEG C of incubators 24 hours.
The chain saturated fatty acids C of table 217H34O3To the growth inhibition effect of phytopathogen
Experimental result is as shown in table 2, the chain saturated fatty acids solution (100 μ g/ml) of separation to cucumber Fusarium oxysporum,
Sclerotium bacteria, banana Fusarium oxysporum, potato Rhizoctonia solani Kuhn, Rhizoctonia cerealis, corn stem rot Fusarium moniliforme and Solanaceae labor
You have very strong growth inhibitory activity by bacterium.
Embodiment 4：Verify growth inhibitory activity of the aliphatic acid of the present invention to staphylococcus aureus：
Two plants of bacterial strains, ACCC NO.01337 Staphylococcus aureus are obtained from DSMZ of Scientia Agricultura Sinica institute
Bacterium and the golden yellow subspecies of ACCC NO.10449 staphylococcus aureuses.First choice uniform sprinkling respectively on the flat board of LB culture mediums
The bacterium solution of the golden yellow subspecies of upper 01337 staphylococcus aureus and 10449 staphylococcus aureuses, after drying, place Oxford cup, ox
Add the chain saturated fatty acids C of 100ul separation in the cup of Tianjin respectively17H34O3Solution (100 μ g/ml).After 4 DEG C stand 8 hours, it is put into
37 DEG C of culture 12h, observation experiment result.
The chain saturated fatty acids C of table 317H34O3To the growth inhibition effect of staphylococcus aureus
Experimental result is as shown in table 3, and the chain saturated fatty acids solution (100 μ g/ml) of separation has to Staphylococcus aureus
Very strong growth inhibitory activity, Staphylococcus aureus is grown completely.
Claims (10)
1. one kind comes from 16 carbon chain fatty acid class antagonistic substances caused by bacillus amyloliquefaciens SQR9, it is characterised in that structure
It is as follows:
2. 16 carbon chain fatty acid class antagonistic substances described in claim 1 are separated in a kind of SQR9 from bacillus amyloliquefaciens
Method, it is characterised in that including：The bacillus amyloliquefaciens SQR9 that fermentative preservation numbering is CGMCC NO.5808, fermentation ends
Afterwards, zymotic fluid is centrifuged, collects supernatant, after moisture removal is removed in supernatant drying, extracted with methanol, concentration leaching liquor obtains medicinal extract；
Silicagel column on medicinal extract, with chloroform-methanol gradient elution, collect chloroform-methanol 20:1 eluent；By chloroform-methanol 20:1 elution
Liquid is dissolved after removing solvent with methanol, further with MPLC ODS chromatographic columns methanol-water gradient elution, collects 80% first
Alcohol-water elution further with rp-hplc method carry out it is deep isolate and purify, in acetonitrile-water 65:35, flow velocity 2mL/min,
Detection wavelength is 210nm, and 16 carbon chain fatty acid class antagonistic substances described in claim 1 are obtained under Rt=12.1.
3. according to the method for claim 2, it is characterised in that the solution starch bud that fermentative preservation numbering is CGMCC NO.5808
Spore bacillus SQR9 fermentation medium is Landy culture mediums.
4. according to the method for claim 2, it is characterised in that bacillus amyloliquefaciens SQR9 fermentation process includes：Press
SQR9 seed liquors are transferred in fresh Landy culture medium conical flasks by 1% inoculum concentration, 37 DEG C, 170rpm, 12h；Again will training
Bacterium solution after supporting is inoculated into 7L small-sized fermentation tank, 5L Landy culture mediums is filled in fermentation tank, 37 DEG C, 200rpm is cultivated
12h；Bacterium solution in 7L fermentation tanks is inoculated into 150L fermentation tanks, 100L Landy culture mediums are filled in fermentation tank, during beginning
Rotating speed is 30 DEG C, rotating speed 200rpm, and oxygen dissolving value 75, defoamer is polyether compound, when oxygen dissolving value starts to drop to below 30,
Rotating speed, up to 500rpm are gradually stepped up, while increases throughput, oxygen dissolving value is maintained at more than 10, until OD600=15,
Rotating speed is now adjusted to 200rpm, throughput is reduced to minimum, and bacterium solution enters stationary phase, starts a large amount of cometabolisms that produce and produces
Thing, ferment 24h.
5. according to the method for claim 2, it is characterised in that described medicinal extract adds methanol to make fully to dissolve, and lysate adds
The mesh silica gel of quality 100~200 makes to be sufficiently mixed, and is concentrated under reduced pressure to be drying to obtain and mixes sample silica gel；Separately weigh the 100 of 2 times of medicinal extract quality
~200 mesh silica gel are placed among glass column as separation silica gel, using chloroform-methanol volume ratio as 1:0；100:1；80:1；
60:1；40:1；20:1 gradient elution.
6. according to the method for claim 2, it is characterised in that gel is passed through to the active component that rp-hplc method is separated to
Chromatogram (85% methanol) is separated except inactive impurity, finally obtains 16 carbon chain fatty acids described in claim 1
The sterling of class antagonistic substance.
7. 16 carbon chain fatty acid class antagonistic substances described in claim 1 are suppressing plant pathogenic fungi and pathogenetic bacteria growth
In application.
8. application according to claim 7, it is characterised in that described pathogenetic bacteria is gram-positive bacterium.
9. application according to claim 7, it is characterised in that 16 carbon chain fatty acid class antagonists described in claim 1
Matter is suppressing cucumber Fusarium oxysporum, sclerotium bacteria, banana Fusarium oxysporum, Phytophthora capsici, potato Rhizoctonia solani Kuhn, cereal silk
Application in pyrenomycetes and Solanaceae Raul bacterium, staphylococcus aureus growth.
10. 16 carbon chain fatty acid class antagonistic substances described in claim 1 suppress plant pathogenic fungi in preparation and cause of disease is thin
Application in the preparation of bacteria growing.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112106775A (en) * | 2019-06-21 | 2020-12-22 | 中国农业大学 | Application of hydroxycarboxylic acid compound in preventing and treating plant diseases |
| CN112898147A (en) * | 2021-02-09 | 2021-06-04 | 中国农业大学 | Method for asymmetric synthesis of (13R,14S) -13-hydroxy-14-methylhexadecanoic acid |
| CN113755394A (en) * | 2021-06-19 | 2021-12-07 | 南京农业大学 | Biological carbon-based microbial fertilizer containing bacillus amyloliquefaciens and application thereof |
| CN114854810A (en) * | 2022-06-17 | 2022-08-05 | 南京农业大学 | Method for improving yield of antibacterial lipopeptide bacillus mycin D through fatty acid metabolism regulation |
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2017
- 2017-09-06 CN CN201710797445.0A patent/CN107501086B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112106775A (en) * | 2019-06-21 | 2020-12-22 | 中国农业大学 | Application of hydroxycarboxylic acid compound in preventing and treating plant diseases |
| CN112106775B (en) * | 2019-06-21 | 2022-04-22 | 中国农业大学 | Application of hydroxycarboxylic acid compound in preventing and treating plant diseases |
| CN112898147A (en) * | 2021-02-09 | 2021-06-04 | 中国农业大学 | Method for asymmetric synthesis of (13R,14S) -13-hydroxy-14-methylhexadecanoic acid |
| CN112898147B (en) * | 2021-02-09 | 2022-02-11 | 中国农业大学 | Method for asymmetric synthesis of (13R,14S) -13-hydroxy-14-methylhexadecanoic acid |
| CN113755394A (en) * | 2021-06-19 | 2021-12-07 | 南京农业大学 | Biological carbon-based microbial fertilizer containing bacillus amyloliquefaciens and application thereof |
| CN114854810A (en) * | 2022-06-17 | 2022-08-05 | 南京农业大学 | Method for improving yield of antibacterial lipopeptide bacillus mycin D through fatty acid metabolism regulation |
| CN114854810B (en) * | 2022-06-17 | 2024-01-26 | 南京农业大学 | Method for improving yield of antibacterial lipopeptide bacilomycin D through fatty acid metabolism regulation |
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