CN103589671A - Bacillus subtilis LNXM37 and application thereof - Google Patents
Bacillus subtilis LNXM37 and application thereof Download PDFInfo
- Publication number
- CN103589671A CN103589671A CN201310561031.XA CN201310561031A CN103589671A CN 103589671 A CN103589671 A CN 103589671A CN 201310561031 A CN201310561031 A CN 201310561031A CN 103589671 A CN103589671 A CN 103589671A
- Authority
- CN
- China
- Prior art keywords
- lnxm37
- subtilis
- application
- microbial preparation
- cgmcc
- 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
- 235000014469 Bacillus subtilis Nutrition 0.000 title claims abstract description 10
- 244000063299 Bacillus subtilis Species 0.000 title claims abstract description 9
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 9
- 235000009566 rice Nutrition 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 235000006008 Brassica napus var napus Nutrition 0.000 claims abstract description 5
- 241000221662 Sclerotinia Species 0.000 claims abstract description 4
- 240000007594 Oryza sativa Species 0.000 claims abstract 3
- 241000209140 Triticum Species 0.000 claims description 13
- 235000021307 Triticum Nutrition 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 10
- 239000000843 powder Substances 0.000 claims description 10
- 201000010099 disease Diseases 0.000 claims description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- 230000000813 microbial effect Effects 0.000 claims description 9
- 244000005700 microbiome Species 0.000 claims description 6
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 4
- 235000015097 nutrients Nutrition 0.000 claims description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 238000000855 fermentation Methods 0.000 claims description 2
- 230000004151 fermentation Effects 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 239000002207 metabolite Substances 0.000 claims description 2
- 230000001737 promoting effect Effects 0.000 claims description 2
- 229940001516 sodium nitrate Drugs 0.000 claims description 2
- 235000010344 sodium nitrate Nutrition 0.000 claims description 2
- 239000004317 sodium nitrate Substances 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 240000000385 Brassica napus var. napus Species 0.000 claims 1
- 230000000593 degrading effect Effects 0.000 abstract description 7
- 239000010902 straw Substances 0.000 abstract description 7
- 240000002791 Brassica napus Species 0.000 abstract description 4
- 241001617088 Thanatephorus sasakii Species 0.000 abstract 1
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 230000000452 restraining effect Effects 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 23
- 230000015556 catabolic process Effects 0.000 description 17
- 238000006731 degradation reaction Methods 0.000 description 17
- 230000001580 bacterial effect Effects 0.000 description 16
- 238000000034 method Methods 0.000 description 11
- 238000012216 screening Methods 0.000 description 9
- 241000193830 Bacillus <bacterium> Species 0.000 description 8
- 241000813090 Rhizoctonia solani Species 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 241000209094 Oryza Species 0.000 description 6
- 241000221696 Sclerotinia sclerotiorum Species 0.000 description 6
- 230000000443 biocontrol Effects 0.000 description 6
- 241000607479 Yersinia pestis Species 0.000 description 5
- 230000000844 anti-bacterial effect Effects 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 229920002488 Hemicellulose Polymers 0.000 description 4
- 239000001913 cellulose Substances 0.000 description 4
- 229920002678 cellulose Polymers 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 229920005610 lignin Polymers 0.000 description 4
- 239000002689 soil Substances 0.000 description 4
- 230000004580 weight loss Effects 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 230000003213 activating effect Effects 0.000 description 3
- 230000008485 antagonism Effects 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 230000003413 degradative effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000003628 erosive effect Effects 0.000 description 3
- 108010002430 hemicellulase Proteins 0.000 description 3
- 229940059442 hemicellulase Drugs 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 2
- 241000193755 Bacillus cereus Species 0.000 description 2
- 241000194108 Bacillus licheniformis Species 0.000 description 2
- 241000194103 Bacillus pumilus Species 0.000 description 2
- 241000193388 Bacillus thuringiensis Species 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 2
- 229940106157 cellulase Drugs 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 235000019387 fatty acid methyl ester Nutrition 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000002255 vaccination Methods 0.000 description 2
- 206010003694 Atrophy Diseases 0.000 description 1
- 241000193410 Bacillus atrophaeus Species 0.000 description 1
- 208000016444 Benign adult familial myoclonic epilepsy Diseases 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241000345369 Edwardsiella hoshinae Species 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 241000589157 Rhizobiales Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000907138 Xanthomonas oryzae pv. oryzae Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 229940097012 bacillus thuringiensis Drugs 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003245 coal Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 244000000037 crop pathogen Species 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 208000016427 familial adult myoclonic epilepsy Diseases 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- ZGNITFSDLCMLGI-UHFFFAOYSA-N flubendiamide Chemical compound CC1=CC(C(F)(C(F)(F)F)C(F)(F)F)=CC=C1NC(=O)C1=CC=CC(I)=C1C(=O)NC(C)(C)CS(C)(=O)=O ZGNITFSDLCMLGI-UHFFFAOYSA-N 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 230000001408 fungistatic effect Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000003345 natural gas Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 235000006180 nutrition needs Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 238000010422 painting Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000000791 photochemical oxidant Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Abstract
The invention discloses Bacillus subtilis LNXM37 which is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation date is September 9 th, 2013, and the preservation number is CGMCC No.8141. The Bacillus subtilis LNXM37 can be used for degrading crop straws, and has a function of restraining phytopathogen such as pellicularia sasakii, sclerotinia rot of colza and the like, so that growth of rice and oilseed rape is promoted, and a good application prospect is achieved.
Description
Technical field
The present invention relates to microbial technology field, relate in particular to a kind of subtilis LNXM37 and application thereof.
Background technology
Crop material is in the world today, to be only second to coal, oil, and the fourth-largest energy of Sweet natural gas, is also unique reproducible energy, is to have one of biomass resource of the potentiality utilized most.In traditional agriculture, stalk resource is directly used in fertilizer, fuel and feed without any processing, utilization ratio is lower, and a large amount of stalks suitablely utilizes method and by peasant's on-site incineration owing to lacking, a kind of waste to resource on the one hand, on the other hand, brought grave danger to surrounding area safety and environment, as discharge the harmful materials such as a large amount of carbon monoxide, oxynitride, photochemical oxidants and suspended particle.
At present, although the method for straw utilization has a lot, also good in the effect of different areas, due to technology and cost, stalk remains the restriction to a certain degree in the application of industry and energy field; In addition, owing to collecting at present, transportation stalk cost is high and purchasing price is low, peasant lacks enthusiasm, thereby does not fundamentally solve stalk problem of outlet, and peasants are still at a large amount of stalks of on-site incineration year in year out.
Just China and even global straw utilization situation, straw-returning is that the current scope of application is the widest, that the most easily promotes utilizes method.Straw-returning can be discharged into land for growing field crops by the C in stalk, N, K, P and trace element, the experimental results shows, straw-returning can effectively improve soil physical property, increase soil nutrient content, be conducive to the accumulation of soil with organic matter, the long-term straw-returning that keeps, can effectively solve current Chinese arable land productivity and fertility decline problem, significantly improves crop yield simultaneously and reduces the even use of Substitute For Partial chemical fertilizer.Yet decomposition speed is slow under field conditions (factors) for the not treated direct returning to farmland of stalk, impact is sowed, is emerged and seedling growth; And can provide hibernaculum to disease and pest after stalk covering, easily make second stubble crop that all kinds of disease and pests occur.
Subtilis (Bacillus subtilis) is a kind of heat-resisting, aerobic G
+rod-shaped bacterium, this bacterium can produce the gemma of strong stress resistance, can significantly suppress pathogenic bacteria, is a kind of common plant growth-promoting rhizobacteria (PGPR), is also in current biocontrol bacteria, to study and apply a more bacterioid; Meanwhile, a large amount of reports also confirm, genus bacillus can produce born of the same parents' outer fiber element enzyme, hemicellulase etc., effectively lignocellulose degradation.Therefore,, in straw degradative process, the Biocontrol Bacillus subtilis that utilization can produce lignocellulose degradation enzyme accelerates the degraded of straw lignocellulose, and nutrient is the most effectively discharged; Meanwhile, not only harmless to field crop itself as the genus bacillus of biocontrol microorganisms, also have the antagonistic action to pathogenic micro-organism concurrently, thereby reduce disease occurrence risk.Subtilis growth and breeding is fast, nutritional needs is low, to person poultry harmless, can suppress various crop disease and produce lipopeptid compound, and its mass production processes is simple, cost is lower, use conveniently, the shelf lives is long, can not allow farm crop develop immunity to drugs, these advantages make it to become a kind of desirable crop material degraded preparation.
Summary of the invention
The object of the invention is to propose subtilis (Bacillus subtilis) LNXM37 of a kind of crop material of can effectively degrading, while energy antagonism crop pathogens.
For reaching this object, the present invention by the following technical solutions:
(Bacillus subtilis) LNXM37, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and preservation date is on September 9th, 2013, and deposit number is CGMCC No.8141.
This bacterium is gram-positive microorganism, shaft-like, without pod membrane, aerobic, and peritrichous can move.Bacterium colony surface irregularity is opaque, micro-yellow.Physio-biochemical characteristics: catalase is positive, V-P reaction is positive, hydrolyzed starch, casein, edwardsiella hoshinae.With upper (the http://www.ncbi.nlm.nih gov/blast/Blast.cgi) known array of 16S rDNA sequencing result and Genbank, compare, with Bacillus subtilis BI1 similarity be 100%.
Comprise described subtilis LNXM37 and/or its meta-bolites and take its microbial preparation that is activeconstituents and also belong to protection scope of the present invention.
Preferably, described microbial preparation is liquid.
The present invention also provides the preparation method of mentioned microorganism preparation, comprises the steps:
(1) described subtilis LNXM37 is inoculated in to LB solid medium, at 28～37 ℃, cultivates and within 14～24 hours, obtain thalline;
(2), in thalline access LB liquid nutrient medium step (1) being obtained, at 28～37 ℃, shaking culture is made seed liquor for 12～24 hours;
(3) inoculum size that by volume per-cent is 5%, is inoculated in step (2) gained seed liquor in fermention medium, in 28～37 ℃ of bottom fermentations, cultivates at least 36～48 hours, collects fermented liquid and obtains described microbial preparation.
In above-mentioned preparation method, preferably, described fermention medium comprises the component of following weight percent: wheat stalk powder 1～3%, preferably 2%, dipotassium hydrogen phosphate 0.1～0.5%, preferably 0.2%, SODIUMNITRATE 0.3～0.8%, preferably 0.5% and glucose 0.5～1.5%, preferably 1%.
The present invention also provides the application of described subtilis LNXM37, and concrete application is as follows:
The application of described subtilis LNXM37 in crop material degraded; Preferably, described crop material is wheat stalk;
The application of described subtilis LNXM37 in preventing and treating rice sheath blight disease;
The application of described subtilis LNXM37 in control sclerotinia rot of colza;
The application of described subtilis LNXM37 in promoting paddy rice and growth of rape.
Subtilis LNXM37 provided by the invention has obvious degradation effect to crop materials such as wheat stalks, and the phytopathogens such as Rhizoctonia solani Kuhn, Sclerotinia sclerotiorum are had to obvious inhibition.Subtilis LNXM37 independent role of the present invention is when stalk, and stalk stalk rate of weight loss can reach 35.2%, and cellulose degradation rate reaches 26.0%, and hemicellulose degradation rate reaches 25.2%, and Lignin degradation rate reaches 48.5%.
Preservation information
Preservation date: on September 9th, 2013;
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center (be called for short: CGMCC), address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, 100101;
Deposit number: CGMCC No.8141.
Accompanying drawing explanation
Fig. 1 is the BOX-PCR finger printing of the part bacterial strain of embodiment 1 separation.
Fig. 2 is the phylogenetic tree based on 16S rDNA sequence construct.
Fig. 3 is to the partial results of the plate screening of 31 strain Biocontrol Bacillus (A), the outer circumscribed cellulase activity (B) of born of the same parents and hemicellulase work (C) measurement result described in embodiment 2.
Fig. 4 is the rate of weight loss curve of subtilis LNXM37 of the present invention wheat stalk powder while degrading separately.
Fig. 5 is the cellulose degradation rate curve of subtilis LNXM37 of the present invention wheat stalk powder while degrading separately.
Fig. 6 is the hemicellulose degradation rate curve of subtilis LNXM37 of the present invention wheat stalk powder while degrading separately.
Fig. 7 is the lignin degradation rate curve of subtilis LNXM37 of the present invention wheat stalk powder while degrading separately.
Fig. 8 has shown the bacteriostatic activity of separated subtilis LNXM37 of the present invention; From left to right be followed successively by the bacteriostatic activity design sketch to Rhizoctonia solani Kuhn (A) and Sclerotinia sclerotiorum (B); Wherein, CK represents blank group, and LNXM37 represents subtilis LNXM37 treatment group of the present invention.
Embodiment
For the present invention is described better, be convenient to understand technical scheme of the present invention, typical but non-limiting embodiment of the present invention is as follows.
Adopt five point sampling ，Yu Lhasa cities, Nyingchi Prefecture, the Shannan Prefecture and Shigatse District, gather altogether 35 soil samples under varying environment condition, to dilute painting flat band method, bacterial strain is carried out to separation and purification, separation obtains 1388 bacterial strains.Then, take rice leaf spot bacteria (Xanthomonas oryzae pv.oryzae), Rhizoctonia solani Kuhn (Rhizoctonia solani) and Sclerotinia sclerotiorum (Sclerotinia sclerotiorum) is indicator, finally obtains the good bacterial strain of 311 strain fungistatic effect (antibacterial circle diameter >=5mm).The isolated strains of these 311 good antimicrobial effect is carried out to genome rep-PCR(repetitive extragenic palindromic PCR) finger printing screening, obtain altogether 150 different bacterial strains of finger printing (Fig. 1 is shown in by finger printing).150 bacterial strains that screening obtains through finger printing are carried out to Physiology and biochemistry evaluation, 150 bacterial strains are divided into 7 monoids, then, from each monoid, choose representational bacterial strain totally 31 strains carry out lipid acid gas-chromatography (fatty acid methyl ester, FAME) and 16S rDNA Molecular Identification (see figure 2), finally, the result of comprehensive phenotypic evaluation and a series of Molecular Identification, by separation, there are 31 Bacillus strains of Biocontrol Potential to be accredited as: bacillus pumilus (B.pumilus) 11 strains, bacillus cereus (B.cereus) 7 strains, bacillus thuringiensis (B.thuringiensis) 3 strains, bacillus amyloliquefaciens (B.amyloliquefaciens) 3 strains, atrophy genus bacillus (B.atrophaeus) 3 strains, B.axarquiensis2 strain, subtilis (B.subtilis) 1 strain, Bacillus licheniformis (B.licheniformis) 1 strain.
The 31 strain Biocontrol Bacillus that the embodiment 1 of take is identified, as basis, carry out first round primary dcreening operation with dull and stereotyped transparent circle method, obtain producing bacterial strain 27 strains of extracellular lignocellulose degrading enzyme, and part plate screening the results are shown in Figure 3A.And then with polysaccharide substrate of different nature, measure the size of this 27 bacillus extracellular enzyme vigor, the outer circumscribed cellulase activity of born of the same parents and hemicellulase measurement result alive are shown in respectively Fig. 3 B and Fig. 3 C.Last comprehensive primary dcreening operation and sieve again result, obtains bacterial strain GBSW19 and LNXM37 that 2 strains have biological and ecological methods to prevent plant disease, pests, and erosion and straw degradative potentiality concurrently.
The actual degradation effect comprehensive test of embodiment 3 bacterial strain LNXM37 to wheat stalk powder
The LNXM37 bacterial strain activating on embodiment 2 gained flat boards is chosen to liquid LB substratum, in 37 ℃, 200rpm shaking culture is made seed liquor for 12 hours, the bacterium amount that connects that the volume percent of take is again 2% is forwarded to adds shaking culture in the Hutchinson substratum that wheat stalk powder (5 order) is sole carbon source, in not collecting on the same day time nutrient solution, and with the poor rate of weight loss that weighs method and improved Van Voest method mensuration wheat stalk powder, cellulose degradation rate, hemicellulose degradation rate and Lignin degradation rate, finally obtain the actual degradation effect comprehensive test result (see Fig. 4-7) of bacterial strain LNXM37 to wheat stalk powder.
Fig. 4-7 result shows: bacterial strain LNXM37 independent role is when stalk, and stalk rate of weight loss can reach 35.2%, and cellulose degradation rate reaches 26.0%, and hemicellulose degradation rate reaches 25.2%, and Lignin degradation rate reaches 48.5%.
The bacteriostatic activity of embodiment 4 subtilis LNXM37 of the present invention
A. the antibacterial screening to pathogenetic bacteria rice leaf spot bacteria
By indicator pathogenetic bacteria rice leaf spot bacteria by a certain percentage (1:30) add in the NA substratum that is cooled to 50 ℃ of left and right and mix, make containing bacterium dull and stereotyped, at 4 filter papers that diameter is 4mm of the vertical placement of distance center 2.5cm place cross, draw bacterial strain bacterium liquid 5 μ l to be measured, be inoculated on filter paper, test bacterium is not inoculated in contrast, with sterilized water, replaces, and each is processed in triplicate.Be placed in 28 ℃ of incubators and cultivate altogether after 2-3 days, observe and record result.
B. the antagonism screening to pathogenic fungi Sclerotinia sclerotiorum
The sclerotinia rot of colza germ PDA plate edge activating in 26 ℃ of incubators is played the bacterium dish that cut-off footpath is 0.7cm (getting the vigorous mycelia piece of indicator marginal growth), is seeded in the dull and stereotyped central authorities of new PDA.Apart from bacterium piece 2.5cm place, be four vaccinations that " ten " font distributes, put the filter paper sequin of diameter 4mm, get bacterium liquid 5ul point to be measured in filter paper central authorities, test bacterium is not inoculated in contrast, with sterilized water, replaces, and each is processed and repeats 3 times, put into 26 ℃ of constant incubators and cultivate after 2-3 days, take out and observe and record antibacterial result.
C. the antagonism screening to pathogenic fungi Rhizoctonia solani Kuhn
The Rhizoctonia solani Kuhn PDA plate edge activating in 28 ℃ of incubators is played the bacterium dish that cut-off footpath is 0.7cm (getting the vigorous mycelia piece of indicator marginal growth), is seeded in the dull and stereotyped central authorities of new PDA.Apart from bacterium piece 2.5cm place, be four vaccinations that " ten " font distributes, put the filter paper sequin of diameter 7mm, get bacterium liquid 5ul point to be measured in filter paper central authorities, test bacterium is not inoculated in contrast, with sterilized water, replaces, and each is processed and repeats 3 times, put into 26 ℃ of constant incubators and cultivate after 3-5 days, take out and observe and record antibacterial result.
Subtilis LNXM37 of the present invention can effectively suppress Rhizoctonia solani Kuhn and Sclerotinia sclerotiorum, and the equal >5mm of its antibacterial circle diameter, as shown in the A of Fig. 8 and B; To rice leaf spot bacteria without obvious inhibition.
Applicant statement, the present invention illustrates the present invention by above-described embodiment, but that the present invention is not limited to is above-mentioned, does not mean that the present invention must rely on above-mentioned could enforcement.Person of ordinary skill in the field should understand, any improvement in the present invention, to the selection of the interpolation of the equivalence replacement of the selected raw material of the present invention and ancillary component, concrete mode etc., within all dropping on protection scope of the present invention and open scope.
Claims (8)
1. subtilis (Bacillus subtilis) LNXM37, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), and preservation date is on September 9th, 2013, and deposit number is CGMCC No.8141.
2. a microbial preparation, is characterized in that, comprises subtilis LNXM37 and/or its meta-bolites described in claim 1.
3. microbial preparation according to claim 2, is characterized in that, described microbial preparation is liquid.
4. the preparation method of microbial preparation as claimed in claim 3, comprises the steps:
(1) described subtilis LNXM37 is inoculated in to LB solid medium, at 28～37 ℃, cultivates and within 14～24 hours, obtain thalline;
(2), in thalline access LB liquid nutrient medium step (1) being obtained, at 28～37 ℃, shaking culture is made seed liquor for 12～24 hours;
(3) inoculum size that by volume per-cent is 5%, is inoculated in step (2) gained seed liquor in fermention medium, in 28～37 ℃ of bottom fermentations, cultivates at least 36～48 hours, collects fermented liquid and obtains described microbial preparation;
Preferably, described fermention medium comprises the component of following weight percent: wheat stalk powder 1～3%, preferably 2%, dipotassium hydrogen phosphate 0.1～0.5%, preferably 0.2%, SODIUMNITRATE 0.3～0.8%, preferably 0.5% and glucose 0.5～1.5%, preferably 1%.
5. the application of subtilis LNXM37 in crop material degraded described in claim 1; Preferably, described crop material is wheat stalk.
6. the application of subtilis LNXM37 in preventing and treating rice sheath blight disease described in claim 1.
7. the application of subtilis LNXM37 in control sclerotinia rot of colza described in claim 1.
8. the application of subtilis LNXM37 in promoting paddy rice and growth of rape described in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310561031.XA CN103589671B (en) | 2013-11-12 | 2013-11-12 | A kind of subtilis LNXM37 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310561031.XA CN103589671B (en) | 2013-11-12 | 2013-11-12 | A kind of subtilis LNXM37 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103589671A true CN103589671A (en) | 2014-02-19 |
| CN103589671B CN103589671B (en) | 2015-08-19 |
Family
ID=50079989
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310561031.XA Active CN103589671B (en) | 2013-11-12 | 2013-11-12 | A kind of subtilis LNXM37 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103589671B (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104762089A (en) * | 2015-04-03 | 2015-07-08 | 桃源县兴隆米业科技开发有限公司 | Selenium-rich soil conditioner for producing natural selenium-rich rice as well as preparation and use methods of selenium-rich soil conditioner |
| CN105018386A (en) * | 2015-07-31 | 2015-11-04 | 长江大学 | Bacillus methylotrophicus and application thereof |
| CN105483053A (en) * | 2016-01-04 | 2016-04-13 | 江西天人生物控股有限公司 | Microbial compound bacteria for preventing and treating citrusyellowshoot and preparation method and application thereof |
| CN105385629B (en) * | 2015-12-01 | 2018-11-30 | 北京德瑞丰农业科技有限责任公司 | One plant of lignocellulosic substance efficient degrading bacteria M2 and its application |
| CN109504632A (en) * | 2018-12-18 | 2019-03-22 | 河北农业大学 | One bacillus subtilis and its application |
| CN109497108A (en) * | 2018-12-25 | 2019-03-22 | 长江师范学院 | A kind of coptis soil-borne disease control agent and preparation method thereof |
| CN109929784A (en) * | 2019-04-15 | 2019-06-25 | 青岛宝创生物科技有限公司 | One bacillus subtilis and its cultural method and application |
| CN111518731A (en) * | 2020-05-14 | 2020-08-11 | 河北环境工程学院 | Bacillus subtilis with antagonistic effect for degrading cellulose at low temperature and application thereof |
| CN113957002A (en) * | 2021-08-26 | 2022-01-21 | 中国林业科学研究院林产化学工业研究所 | Lignin degrading bacillus and application thereof |
| CN114591857A (en) * | 2022-03-10 | 2022-06-07 | 内蒙古工业大学 | Bacillus subtilis with high fiber degradation rate and application thereof |
| CN114806926A (en) * | 2022-03-22 | 2022-07-29 | 扬州大学 | Kitchen waste decomposing and degrading microbial inoculum and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401587A (en) * | 2008-11-21 | 2009-04-08 | 天津科技大学 | Bacillus subtilis formulation for preventing and controlling soil-borne disease and method of preparing the same |
| CN102344320A (en) * | 2011-07-19 | 2012-02-08 | 杨凌绿都生物科技有限公司 | Novel high-efficiency biocontrol bacterial fertilizer and production technology thereof |
| CN102517237A (en) * | 2011-12-30 | 2012-06-27 | 湖南农业大学 | Composite micro-ecological water-based suspension concentrate for aquaculture |
| CN103173394A (en) * | 2013-03-21 | 2013-06-26 | 西南大学 | Bacillus subtilis for controlling citrus canker |
-
2013
- 2013-11-12 CN CN201310561031.XA patent/CN103589671B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401587A (en) * | 2008-11-21 | 2009-04-08 | 天津科技大学 | Bacillus subtilis formulation for preventing and controlling soil-borne disease and method of preparing the same |
| CN102344320A (en) * | 2011-07-19 | 2012-02-08 | 杨凌绿都生物科技有限公司 | Novel high-efficiency biocontrol bacterial fertilizer and production technology thereof |
| CN102517237A (en) * | 2011-12-30 | 2012-06-27 | 湖南农业大学 | Composite micro-ecological water-based suspension concentrate for aquaculture |
| CN103173394A (en) * | 2013-03-21 | 2013-06-26 | 西南大学 | Bacillus subtilis for controlling citrus canker |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104762089A (en) * | 2015-04-03 | 2015-07-08 | 桃源县兴隆米业科技开发有限公司 | Selenium-rich soil conditioner for producing natural selenium-rich rice as well as preparation and use methods of selenium-rich soil conditioner |
| CN104762089B (en) * | 2015-04-03 | 2017-11-14 | 桃源县兴隆米业科技开发有限公司 | A kind of selenium-rich soil modifying agent for producing natural selenium-rich rice and its preparation, application method |
| CN105018386A (en) * | 2015-07-31 | 2015-11-04 | 长江大学 | Bacillus methylotrophicus and application thereof |
| CN105385629B (en) * | 2015-12-01 | 2018-11-30 | 北京德瑞丰农业科技有限责任公司 | One plant of lignocellulosic substance efficient degrading bacteria M2 and its application |
| CN105483053A (en) * | 2016-01-04 | 2016-04-13 | 江西天人生物控股有限公司 | Microbial compound bacteria for preventing and treating citrusyellowshoot and preparation method and application thereof |
| CN105483053B (en) * | 2016-01-04 | 2018-12-11 | 江西天人生物控股有限公司 | A kind of microbial composite bacteria and its preparation method and application for preventing and treating Citrus Huanglongbing pathogen |
| CN109504632B (en) * | 2018-12-18 | 2022-02-25 | 河北农业大学 | Bacillus subtilis and application thereof |
| CN109504632A (en) * | 2018-12-18 | 2019-03-22 | 河北农业大学 | One bacillus subtilis and its application |
| CN109497108A (en) * | 2018-12-25 | 2019-03-22 | 长江师范学院 | A kind of coptis soil-borne disease control agent and preparation method thereof |
| CN109929784A (en) * | 2019-04-15 | 2019-06-25 | 青岛宝创生物科技有限公司 | One bacillus subtilis and its cultural method and application |
| CN111518731A (en) * | 2020-05-14 | 2020-08-11 | 河北环境工程学院 | Bacillus subtilis with antagonistic effect for degrading cellulose at low temperature and application thereof |
| CN111518731B (en) * | 2020-05-14 | 2021-10-19 | 河北环境工程学院 | Bacillus subtilis with antagonistic effect for degrading cellulose at low temperature and application thereof |
| CN113957002A (en) * | 2021-08-26 | 2022-01-21 | 中国林业科学研究院林产化学工业研究所 | Lignin degrading bacillus and application thereof |
| CN114591857A (en) * | 2022-03-10 | 2022-06-07 | 内蒙古工业大学 | Bacillus subtilis with high fiber degradation rate and application thereof |
| CN114591857B (en) * | 2022-03-10 | 2023-05-09 | 内蒙古工业大学 | Bacillus subtilis with high fiber degradation rate and application thereof |
| CN114806926A (en) * | 2022-03-22 | 2022-07-29 | 扬州大学 | Kitchen waste decomposing and degrading microbial inoculum and application thereof |
| CN114806926B (en) * | 2022-03-22 | 2023-08-01 | 扬州大学 | Kitchen waste decomposing degradation microbial inoculum and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103589671B (en) | 2015-08-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103589671B (en) | A kind of subtilis LNXM37 and application thereof | |
| CN103589670B (en) | A kind of Bacillus pumilus GBSW19 and application thereof | |
| CN105524858B (en) | A kind of high temperature resistant decomposing microbial inoculum of organic waste and its application | |
| Zhang et al. | Algal sludge from Taihu Lake can be utilized to create novel PGPR-containing bio-organic fertilizers | |
| CN104017744A (en) | Preparation method and application of pseudomonas chlororaphis for resisting disease and promoting growth | |
| CN101519639B (en) | Paenibacillus polymyxa for preventing and treating plant fungal diseases and production method thereof | |
| CN101497541B (en) | Efficient disease-resistant phosphate solubilizing bacterial manure for tobacco and production method thereof | |
| Zou et al. | Emerging technologies of algae‐based wastewater remediation for bio‐fertilizer production: a promising pathway to sustainable agriculture | |
| CN103224904B (en) | Rhodopseudomonas strain, biocontrol microbial inoculum and biocontrol fermentation liquid, and corresponding preparation methods and application thereof in controlling phytophthora blight of pepper | |
| CN107142210A (en) | A kind of compound method of hard stalk fermentation microbial inoculum | |
| Borah et al. | Low energy rice stubble management through in situ decomposition | |
| Liu et al. | Isolation and evaluation of the plant growth promoting rhizobacterium Bacillus methylotrophicus (DD-1) for growth enhancement of rice seedling | |
| CN103468591B (en) | Salt-tolerant trichoderma pleuroticola strain and application thereof | |
| CN101955903B (en) | Bacillus licheniformis bacterial strain and application thereof | |
| Wu et al. | Rhizobacteria strain from a hypersaline environment promotes plant growth of Kengyilia thoroldiana | |
| Adelere et al. | Valorization of feather by Bacillus safensis and Aquamicrobium defluvii for growth promotion in leafy vegetables | |
| CN105176893B (en) | A kind of bacillus amyloliquefaciens and its application for preventing crop damping-off | |
| CN104560817B (en) | Thermophilic bacillus licheniformis UTM102 for producing phytase and application of thermophilic bacillus licheniformis UTM102 | |
| CN110066748A (en) | A kind of complex microorganism and microbial inoculum and its application comprising the complex microorganism | |
| CN109182219A (en) | One plant of Mo Haiwei bacillus for promoting the growth of Henry David Thoreau grass and its application | |
| CN108676760A (en) | A kind of organic fertilizer decomposing agent and preparation method thereof | |
| CN102925394A (en) | Endophytic bacillus subtilis with disease prevention and growth promotion effects | |
| CN115968910A (en) | Chaetomium globosum DX-THS3 bacterial fertilizer, preparation method and application thereof | |
| CN116121082A (en) | Smoke tube bacterium MR-8 with drought resistance function and application thereof | |
| CN106609250A (en) | Preparation method of fermented traditional Chinese medicine residue organic fertilizer inoculant and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190313 Address after: 211103 2nd Building, 318 Runfa Road, Dongshan Industrial Concentration Zone, Jiangning District, Nanjing City, Jiangsu Province Patentee after: NANJING HOUJI BIO-TECHNOLOGY Co.,Ltd. Address before: 214192 Xiangyun No. 99 Furong Zhongsan Road, Xishan Economic Development Zone, Wuxi City, Jiangsu Province Patentee before: WUXI YAKE BIO-TECHNOLOGY Co.,Ltd. |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20240411 Address after: Room 308-30, Building A, No. 159 Chengjiang Middle Road, Jiangyin City, Wuxi City, Jiangsu Province, 214400 Patentee after: Wuxi Yuansheng Biotechnology Co.,Ltd. Country or region after: China Address before: 211103 2nd Building, 318 Runfa Road, Dongshan Industrial Concentration Zone, Jiangning District, Nanjing City, Jiangsu Province Patentee before: NANJING HOUJI BIO-TECHNOLOGY Co.,Ltd. Country or region before: China |