CN109182219A - One plant of Mo Haiwei bacillus for promoting the growth of Henry David Thoreau grass and its application - Google Patents

One plant of Mo Haiwei bacillus for promoting the growth of Henry David Thoreau grass and its application Download PDF

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CN109182219A
CN109182219A CN201811222398.8A CN201811222398A CN109182219A CN 109182219 A CN109182219 A CN 109182219A CN 201811222398 A CN201811222398 A CN 201811222398A CN 109182219 A CN109182219 A CN 109182219A
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growth
bacillus
kkdd
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bacterial strain
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CN109182219B (en
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谢永丽
吴晓晖
陈兰
柴树芳
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Qinghai University
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    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C21/00—Methods of fertilising, sowing or planting
    • A01C21/005—Following a specific plan, e.g. pattern
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

Abstract

The present invention discloses the Mo Haiwei bacillus and its application that one plant promotees the growth of Henry David Thoreau grass, bacterial strain KKDD belongs to Mo Haiwei bacillus (Bacillus mojavensis), China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on April 11st, 2018, culture presevation number is CGMCC NO.15579, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.It is isolated from Qinghai Province HOh Xil saline-alkali land vegetation rhizosphere bacillus KKDD to have both fixed nitrogen, antagonism pathogen, produce IAA, vernalization growth-promoting and saline-alkali tolerant characteristic, there is certain research and application potential in the recovery of plateau grassland vegetation, ecological countryside.The KKDD bacterial strain also has the function of that degrading straw lignocellulosic, antagonism Henry David Thoreau grass generate fungal disease other than significantly improving the effect of Henry David Thoreau grass growth.

Description

One plant of Mo Haiwei bacillus for promoting the growth of Henry David Thoreau grass and its application
Technical field
The invention belongs to biological pesticide technical field, in particular to one plant of Mo Haiwei Bacillus strain and its in prevention and treatment shuttle Application in sieve grass fungal disease, rush Henry David Thoreau grass growth.
Background technique
Henry David Thoreau grass is grass family Tribe Triticeae Kengyilia herbaceos perennial, and in China, Henry David Thoreau grass integrated distribution is in blueness The ground such as sea, Tibet, are the endemic species of Qinghai-Tibet Platean, and Qinghai Province's Henry David Thoreau grass is mainly distributed on three river sources areas with cooling High aititude.Shuttle Sieve grass solid-holding capacity is strong, and cold-resistant is drought-enduring, anti-blown sand, saline-alkali tolerant, is adapted to the severe high-cold steppe habitat of weather conditions, is high and cold One of the main constituents of steppe plant group, can be used for ecological recovery.Since Henry David Thoreau grass is being rich in nutrition and is easy to animal Digestion, can be used as feed, thus alleviate autumn and winter Qinghai-Tibet Platean grassland it is withered and yellow after, the phenomenon of feed deficiency.
Qinghai-Tibet height above sea level, temperature are low, ultraviolet strong, and special habitats make the universal slow growth of Alpine forage.In recent years, by It endangers, overgraze in mouse worm, the factors such as scientific management are lacked to grassland, good forage is caused to degenerate, malicious weeds propagation and widespread, aggravation Alpine meadow turf peels off, soil bareness, the deterioration of the ecological environment.The formation on degeneration meadow directly threaten this area herdsman and The survival and development of domestic animal also threatens the ecological balance of sources of three rivers In The Middle And Lower Reaches.Therefore, accelerate herbage growth, restore ecology Vegetation improves biological content, particularly important in plateau grassland ecological environment restores and administers.
It can play stably effect in Qinghai-Tibet Alpine meadow extreme environment at present, and be directed to and promote the growth of Henry David Thoreau grass Biocontrol bacterial strain not yet have been reported that.Secondly, a large amount of uses of artificial synthesized pesticide, chemical fertilizer are while improving crop yield to people Class health and ecological environment bring negative effect.The long-time service of chemical fertilizer can bring residual and pollution to Alpine meadow, and poison is miscellaneous Careless largely to breed, harmful organism generates resistance, and injury non-target organism even destruction plateau ecology balance etc. is a series of serious The problem of.Bacillus can generate the inverse gemma of heat-resistant, be conducive to the production and processing of biocontrol agent, and application is convenient, storage Phase is long, becomes the best competitor of chemical fertilizer, pesticide and other probiotics.It is mostly important to be, existing biocontrol bacterial strain Pseudomonas is excessively single, is concentrated mainly on bacillus subtilis and bacillus amyloliquefaciens, if there are other resistance to extreme environments and The biocontrol bacterial strain for efficiently promoting the growth of Henry David Thoreau grass, is the main contents of this research.Mo Haiwei bacillus (Bacillus It mojavensis) is a kind of aerobic sporiferous Gram-positive bacillus for biting warm nature, being able to suppress influences the life of Henry David Thoreau grass Long pathogen will reduce the two packing spaces of environmental pollution caused by chemical pesticide, raising Pasture ecosystem if reasonable utilization Property, keep the Alpine meadow ecological balance.
Summary of the invention
The purpose of the present invention is to provide one kind to adapt to Qinghai-Tibet special habitats, herbage such as Henry David Thoreau grass is promoted to grow, Improve the Mo Haiwei bacillus KKDD of Alpine meadow biomass.
Another object of the present invention is to provide a kind of microbial inoculum and microbial manure comprising above-mentioned bacterial strains.
Of the invention has one to be designed to provide above-mentioned Mo Haiwei bacillus KKDD answering in promotion herbage growth With.
The purpose of the present invention can be achieved through the following technical solutions:
The Mo Haiwei bacillus of one plant of rush herbage growth, bacterial strain KKDD belong to Mo Haiwei bacillus (Bacillus Mojavensis), it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 11st, 2018, Culture presevation number is CGMCC NO.15579, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.The KKDD bacterial strain removes Outside the effect for significantly improving the growth of Henry David Thoreau grass, also there is degrading straw lignocellulosic, antagonism Henry David Thoreau grass to generate fungal disease Effect.
A kind of microbial inoculum promoting herbage growth, includes above-mentioned Mo Haiwei bacillus in the microbial inoculum.
A kind of microbial-bacterial fertilizer promoting herbage growth, includes above-mentioned Mo Haiwei bacillus in the microbial-bacterial fertilizer.
Above-mentioned Mo Haiwei bacillus is preparing the application in the microbial-bacterial fertilizer for promoting herbage growth.
Above-mentioned Mo Haiwei bacillus, microbial inoculum or microbial-bacterial fertilizer is promoting herbage growth, lignocellulose degradation or short of money Application in anti-Fungal Forage Diseases.
A method of promoting herbage growth, be at least one of following (1)~(3):
(1) seed soaking is carried out to grass seed using the fermentation liquid of above-mentioned bacterial strains；
(2) root irrigation is carried out to herbage seedling using the fermentation liquid of above-mentioned bacterial strains；
(3) it is applied fertilizer using above-mentioned microbial-bacterial fertilizer to herbage.
Above-mentioned herbage is preferably Henry David Thoreau grass.
Beneficial effects of the present invention
Using 16S-rDNA and gyrB gene amplification analysis to separation screening from Qinghai Province HOh Xil salt-soda soil Androsace umbellata Spraying potassium KKDD carries out Molecular Identification, and bacterial strain KKDD is accredited as Mo Haiwei bacillus mojavensis, the bacterium Strain salt tolerant alkali ability is stronger, remains to normal growth under conditions of salt content is 11%, pH=11.It is examined by plate dual test Survey the activity of the bacterial strain antagonizing pathogenic fungi, strains on plant disease fungus fusarium graminearum (FusaHum Graminearum), sharp top Fusariumsp (Fusarium acuminatum) has significant antagonistic activity, average diameter of inhibition zone >=15mm.IAA ability is generated by Salkowski colorimetric determination bacterial strain, strains expressed, which goes out, relatively produces by force IAA ability, secretion Amount is 19.904 μ g/mL, meanwhile, strains expressed goes out stable nitrogen fixing capacity.It is soaked seed with bacterial strain fermentation liquor to grass seed, The vernalization and growth-promoting effect of strains on plant are detected, Henry David Thoreau is carelessly planted in the processing of bacterial strain seed soaking has stronger vernalization growth-promoting Effect, compared with the control, germination percentage increase by 8%, and the processing of bacterial strain fermentation liquor method for soaking seed is 7.62% to herbage growth-promoting rate；With bacterial strain The fermentation liquid Henry David Thoreau grass seedling uniform to growing way carries out root irrigation, and growth-promoting rate is 24.60%.It is isolated from Qinghai Province HOh Xil Saline-alkali land vegetation rhizosphere bacillus KKDD has both fixed nitrogen, antagonism pathogen, produces IAA, vernalization growth-promoting and saline-alkali tolerant characteristic, in height Former grassland vegetation restores, has certain research and application potential in ecological countryside.
Detailed description of the invention
The colonial morphology of Fig. 1 bacterial strain KKDD
The effect of Fig. 2 bacterial strain KKDD suppression Henry David Thoreau grass disease fungus
A fusarium graminearum (Fusarium graminearum), the sharp top Fusariumsp of B (Fusarium acuminatum)
The produced lipopeptide compound of Fig. 3 bacterial strain KKDD
The AMALDI-TOF-MS mass spectrogram for the lipopeptid compound that A bacterial strain KKDD is generated
B blood plate transparent circle method detects the active result figure of lipopeptide antibiotic that bacterial strain KKDD is generated
Fig. 4 bacterial strain promotees the growth of Henry David Thoreau grass
Specific embodiment
Embodiment 1:
The separation of 1.1 bacterial strains
From the Qinghai Province HOh Xil Hai Xizhou, (height above sea level: 5500m, year samming is -10.0 DEG C~4.1 DEG C to applicant, average annual to drop Water obtains one plant for separation screening in 173~495mm) salt-soda soil Androsace umbellata (Androsace umbellata) rhizosphere soil Bacterial strain is named as KKDD.Bacterial strain KKDD's of the invention isolates and purifies using the side such as dilution spread flat band method, plate streaking Method.
The identification of 1.2 bacterial strains
1.2.1 the Morphological Identification of bacterial strain
By observation colonial morphology, color, edge, transparency etc., bacterial strain KKDD is cultivated at LB (Luria-Bertan) The scribing line culture of base upper flat plate.As shown in Figure 1, the Main Morphology characteristic of bacterial strain KKDD are as follows: the optimal growth on LB culture medium Temperature is 25 DEG C, after culture 1~2 day, forms the bacterium colony of 2~3cm of diameter, bacterium colony is creamy white, and opaque, colony edge is not whole Together, there is gauffer；In LB liquid medium, 25 DEG C of overnight shaking table cultures, thalli growth is vigorous, highly turbid, and it is cotton-shaped heavy to generate It forms sediment, there is special odor.
1.2.2 the Molecular Identification of bacterial strain
It extracts bacterial genomes DNA and the general of bacterium is designed according to the conservative series at the both ends bacterial 16 S rDNA and gyrB Primer, 16S-27F (5 '-AGAGTTTGATCMTGGCTCAG-3 '), 16S-1492R (5 '-GCYTACCTTGTTACGACTT- 3')；GyrB-For (5 '-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3 '), gyrB-Rev (5 '-AG CAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3').PCR amplification condition are as follows: 95 DEG C of 4min；98℃ 10sec, 62 DEG C of 1min, 72 DEG C of 2min, totally 30 recycle；72℃8min.PCR reaction product is through 1.5% agarose gel electrophoresis Detection after detection is qualified, is sent to the sequencing of Shenzhen Hua Da.
Using bacterial strain KKDD genomic DNA is template amplification 16S rDNA (as shown in SEQ ID NO.1) and gyrB gene is (such as Shown in SEQ ID NO.2), the PCR signature band of size about 1500bp, 1300bp are arrived in amplification respectively, with bacillus 16S The theoretical value of rDNA, gyrB are consistent substantially.Sequencing result is carried out BLAST with known array in GenBank in NCBI to compare. Comparison result shows: bacterial strain KKDD and Bacillus mojavensis strain LMB3G43 sequence (accession number: MF040277.1) identical (or consistent) property is 100%；Bacterial strain KKDD and Bacillus mojavensis strain 28-1 (is stepped on Record number: identical (or consistent) property of gyrB sequence KF194271.1) is 99%.
Identified, bacterial strain KKDD belonged to Mo Haiwei bacillus (Bacillus mojavensis), and in 2018 4 It is preserved within 11st China Committee for Culture Collection of Microorganisms's common micro-organisms center the moon, culture presevation number is CGMCC NO.15579。
The physiological and biochemical test of 1.3 bacterial strain KKDD
1.3.1 resistance to acid and alkali measures:
Draw 100 μ L cultivate to the KKDD bacteria suspension of logarithmic phase be added 10mL pH be respectively 3.0,5.0,7.0,9.0, 11.0, in 13.0 LB liquid medium, training 3d is shaken in 37 DEG C, 180r/min constant temperature, measures its OD600Value is (with LB Liquid Culture Base is control), it is repeated 3 times.Bacterial strain KKDD is not grown under strong acid environment of the pH less than 5, with the reduction of acidity, is gradually become (pH 7~11) thalli growth is gradually vigorous when alkaline environment, and when pH is in strong alkali environment between 11~13, thalli growth is in Downward trend.Thallus at 7~11 pH can normal growth, this optimal pH (6.5~7.5) range compared with most of bacteriums It is wider.
1.3.2 Salt resistant test:
Draw 100 μ L cultivate to the KKDD bacteria suspension of logarithmic phase be added 10mL NaCl concentration be respectively 0,3%, 5%, 7%, in 9%, 11%, 13% LB liquid medium, training 3d is shaken in 37 DEG C, 180r/min constant temperature, measure its OD600Value (with LB liquid medium is control), it is repeated 3 times.Measurement result shows that bacterial strain KKDD has very strong salt resistance ability, is in salt content When growing in 5% LB culture medium, the concentration of thallus is maximum, with the increase of salt content, when salt content is more than 11%, thallus Slow growth, cell concentration are on a declining curve.Thallus can be grown in the environment that salt content is 3%-11%, show bacterial strain The more most of bacterium salt tolerant wider range of salt tolerance.
1.3.3 low temperature resistant measurement:
On LB culture medium flat plate, in four vaccinations of " ten " font distribution, diameter 4mm filter paper sequin is placed, is inhaled 5 μ L are taken to cultivate to the KKDD bacterial suspension inoculation of logarithmic phase in filter paper, each processing is repeated 3 times, and culture medium is placed in 4 DEG C, 10 DEG C, 25 DEG C, cultivate under the conditions of 37 DEG C, observe and measure upgrowth situation of the bacterial strain on culture medium flat plate.Measurement result shows: bacterium Strain KKDD is resistant to 4 DEG C -10 DEG C of low temperature, at 25 DEG C, grows the most vigorous, while can also grow in 37 DEG C of environment.
1.3.4 antibiotic-resistant marks:
By bacterial strain KKDD be inoculated in respectively containing 50 μ g/mL rifampins, kanamycins LB culture medium on, in 25 DEG C of culture 5d Afterwards.The good mutant strain of upgrowth situation is picked out, is transferred to containing rifampin, kanamycins concentration difference 50 μ g/mL, 100 μ g/ ML, 200 μ g/mL, 300 μ g/mL, 400 μ g/mL, 500 μ g/mL LB culture medium in cultivate, be repeated 5 times.Measurement result shows: Bacterial strain KKDD being capable of the rifampin of 50 μ g/mL of antagonism and the kanamycins of 50 μ g/mL.
1.3.5 nitrogen fixing capacity measures:
On Ashby nitrogen-free agar plate, in four vaccinations of " ten " font distribution, it is small to place diameter 4mm filter paper Disk draws 5 μ L at 37 DEG C, the bacterial suspension inoculation of 14h is cultivated under the conditions of 200r/min in filter paper, each processing is repeated 3 times It is repeated 3 times, 37 DEG C of constant temperature incubations, observes and measures upgrowth situation of the bacterial strain on culture medium flat plate.It is trained according to bacterial strain in fixed nitrogen The bacterium colony size formed on base is supported, determines the nitrogen fixing capacity of bacterial strain KKDD, bacterium colony is bigger, indicates that bacterial strain nitrogen fixing capacity is stronger.Bacterium Strain KKDD forms apparent bacterium colony on Ashby nitrogen-free agar, and remains to form obvious bacterium colony after 3 generations continuously cultivated, In Ashby fluid nutrient medium after 3 generation of squamous subculture, bacterium solution is still muddy.
1.3.6 heteroauxin determination of activity is produced:
Using Salkowski colorimetric method, color solution (30mL 0.5mol/L FeCl is added3+ 30mL distilled water+50mL 98%H2SO4) after 15min observe color change, 3 repetitions redden as the positive, and expression can secrete heteroauxin IAA, color It more deeply feels and shows that secretion quantity is more；3 repetitions do not change color as feminine gender, and heteroauxin IAA is not secreted in expression；It is bent using IAA standard Line production will cultivate the bacterium suspension and control centrifugation (4 DEG C, 10000r/min, 10min) of 7d, supernatant 4mL taken to add same-size ratio Color liquid, dark measure OD after standing 30min530, it is repeated 3 times.Bacterial strain KKDD secretion heteroauxin is calculated according to standard curve Amount.
In the LB culture solution containing 100mg/L tryptophan, the color solution that the bacterium is added reddens.It is made and is marked of IAA Directrix curve.Normal equation are as follows: y=19.057x+0.5991, KKDD are in the LB culture solution containing tryptophan, the OD of color solution530 It is 1.013, is according to the secretory volume that normal equation can calculate the KKDD in the Jin Shi culture medium of the tryptophan containing 100mg/L 19.904μg/mL。
The measurement of 1.4 bacterial strain KKDD antagonistic activities
1.4.1 bacterial strain KKDD restraining epiphyte effect measuring
Respectively will in 26 DEG C of incubators activated fusarium graminearum (FusaHum graminearum), sharp top sickle The bacterium dish for taking diameter to be 0.7cm is played at the PDA plate edge spore bacterium (Fusarium acuminatum), is seeded in new PDA plate Centre.At fungus block 2.5cm, in four vaccinations of " ten " font distribution, diameter 4mm filter paper sequin is placed, draws 5 μ L At 37 DEG C, the bacteria suspension of 14h is cultivated under the conditions of 200r/min, is inoculated in filter paper, each processing is repeated 3 times, and is put into 26 DEG C of perseverances After cultivating 2~3d in warm incubator, takes out and observe and record antibacterial result.As shown in Fig. 2, detecting bacterium by plate dual test Strain antagonistic activity, bacterial strain KKDD is to plant pathogenic fungi fusarium graminearum (Fusarium graminearum) as the result is shown (such as Fig. 2A), sharp top Fusariumsp (Fusarium acuminatum) (such as Fig. 2 B) have significant antagonistic activity, and inhibition zone is average Diameter >=15mm, show significant restraining epiphyte effect.
Table 1 antagonizing pathogenic fungi activity
Table 1Antagonistic activity to pathogenic fungus
Note :+: inhibition zone radius 0-5mm；++: antibacterial circle diameter 5-10mm；+++: antibacterial circle diameter 10-15mm；
++++: antibacterial circle diameter > 15mm.
Note:+:Inhibition zone radius 0-5mm；++:Inhibition zone radius 5-10mm； +++:Inhibition zone radius 10-15mm；++++:Inhibition zone radius≥15mm.
1.4.2 the MALDI-TOF-MS mass spectroscopy of bacterial strain KKDD
Biocontrol bacterial strain B.mojavensis KKDD extracts and obtains lipopeptid compound after fermenting by Landy, to lipopeptid It closes object and carries out MALDI-TOF-MS analysis, detect the type and composition of the producible lipopeptid compound of bacterial strain.As shown in Figure 3A, it ties Fruit shows: B.mojavensis KKDD m/z value be 1485.790,1499.799 at have quasi-molecular ions (cluster) appearance, this 2 from Sub- peak both corresponds to the quality of Fengycin；It is to there is quasi-molecular ions (cluster) to go out at 1030.83,1044.75,1058.75 in m/z value Existing, this 3 quasi-molecular ions both correspond to the quality of Surfactin.MALDI-TOF-MS mass spectrogram analysis shows that bacterial strain KKDD can be produced Raw lipopeptide compound Fengycin, Surfactin.
1.4.3 the lipopeptide antibiotic suppression blood plate experiment of bacterial strain KKDD
As shown in Figure 3B, the activity for the lipopeptide antibiotic that strains tested generates, knot are detected by blood plate transparent circle method It is the saturating of 17mm that fruit, which shows that the lipopeptid compound crude extract that bacterial strain B.mojavensis KKDD is extracted forms diameter on blood plate, Bright circle, the lipopeptid compound for showing that bacterial strain generates all have antibiotic activity, can be by dividing with the phosphatide pair on erythrocyte membrane Sublayer interacts to form ion channel, destroys membrane structure, discharges cellular content, causes haemocyte dead.
1.5 bacterial strain KKDD promote the growth of Henry David Thoreau grass
1.5.1 bacterial strain KKDD promotees the sprouting of Henry David Thoreau grass seed seedling
Experiments have shown that by Henry David Thoreau grass seed through bacterial strain KKDD fermentation liquid seed soaking 2h (effective bacterial content are as follows: 1 × 107cfu/ ML), after cultivating 5d, measurement Biocontrol Bacillus promotees Henry David Thoreau grass seed and sprouts effect.The result shows that: strains tested KKDD fermentation liquid Henry David Thoreau has carelessly been planted and has significantly promoted to sprout effect, compared with the control, the Henry David Thoreau grass seed germination percentage of fermented liquid processing is 92%, control group germination percentage is 80%, and germination percentage increases by 12% compared with the control group.
1.5.2 bacterial strain KKDD promotees Henry David Thoreau grass growth of seedling
By Henry David Thoreau grass seed through bacterial strain KKDD fermentation liquid seed soaking 2h (effective bacterial content are as follows: 1 × 107Cfu/mL), 15d is cultivated Afterwards, growth-promoting effect of the measurement Biocontrol Bacillus to Henry David Thoreau grass seedling.As shown in Figure 4, the results showed that: strains tested KKDD bacterium is outstanding Liquid has apparent growth-promoting effect to Henry David Thoreau grass seedling, and compared with the control, the average plant height of the Henry David Thoreau grass seedling through handling reaches 12.15cm increases by 7.62% compared with the control group, and plant height conspicuousness improves.By Henry David Thoreau grass seed through 20% liquor natrii hypochloritis Seed is aseptically sown into the basin of cave by middle disinfection treatment, when plant plant height reaches 5-6cm, with concentration be 1 × 107Cfu/mL ferments liquid irrigating root bacterial strain, growth-promoting effect of the measurement bacterial strain to Henry David Thoreau grass seedling after 10d.The result shows that: strains tested KKDD bacteria suspension has apparent growth-promoting effect to Henry David Thoreau grass seedling, increases by 24.60% compared with the control group, Henry David Thoreau grass plant height is significant Property improve.Compare through many experiments, method for soaking seed growth-promoting effect is better than root-pouring method, and bacterial strain KKDD is promoting Henry David Thoreau grass growth of seedling Meanwhile also improving fresh weight, dry weight and the chlorophyll content of Henry David Thoreau grass.
2 bacillus of table promotees plant seedling growth efficiency
Table 2The effect of plant seedling promoted by Bacillus spp.(Seed soaking)
3 bacterial strain KKDD of table promotees plant seedling growth efficiency (root-pouring method)
Table 3The effect of plant seedling promoted by KKDD(Root irrigation)
16S sequence (SEQ ID NO.1):
CAGATGGGAGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAG ACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGCTTGTTTGAACCGCATGGTTCAAACATAAAAGGTGGCT TCGGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCAACGATGCGT AGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAA TCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTG TTAGGGAAGAACAAGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTG CCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTCCTT AAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAG TGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTA ACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTG CTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAA GACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAA CCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCA TGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGC ATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCC TTATGACCTGGGCTACACACGTGCTACAATGGACAGAACAAAGGGCAGCGAAACCGCGAGGTTAAGCCAATCCCACA AATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCAT GCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTG AGGTAACCTTTATGGAGCCAGCCGCCGAA
GyrB sequence (SEQ ID NO.2):
CGGGCTTTTTCCAAGTTGACATCTTCCCCGATACCTGTGCCGAGTGCAGTAATCATAGAACGAACTTCG TTGTTAGAAAGAATTTTATCAAGCCTTGCTTTTTCAACGTTTAGGATTTTACCTCTTAGCGGCAAAATGGCTTGGAA ATGCCTGTCACGTCCCTGTTTTGCAGATCCTCCGGCAGAGTCACCCTCTACGATATATAACTCGGAGATGCTCGGGT CTTTCGAAGAGCAGTCCGCTAATTTACCGGGAAGGTTTGAAATCTCCAAAGCGCTTTTGCGGCGCGTTAATTCACGC GCTTTTTTCGCAGCCATTCTTGCTCTTGCTGCCATTAAACCTTTGTCAACGATTTTTTTTGCCGCATCTGGATTTTC CAGCATAAATGTTTCCAACGCCGCAGAAAATAACGTATCTGTGATCGTCCGCGCCTCTGAGTTGCCTAGTTTTGTTT TCGTTTGGCCTTCGAACTGCGGATCCGGGTGTTTGATCGAGATAATCGCGGTAAGCCCTTCTCTCACATCATCTCCG CTTAAGTTTGGATCATTTTCTTTTATGAGTCCTTTTTTTCTGGCGTAATCATTGATGACACGAGTCAGCCCCGTTTT AAAACCGGCTTCGTGGGTACCGCCTTCGTACGTGTTGATATTGTTTGTAAATGAGTAAATATTGCTTGTGTAGCTGT CATTGTATTGCAGAGCGACTTCAACCGTAATGCCGTCCTTTTCGCCTTCAATATAAATCGGCTCTTCATGGACAACT TCTTTGGAGCGGTTTAAATACTCTACATAGCTTTTAATTCCGCCTTCGTAATGATACTCATTTTTGCGCTCTTGGCC TTCACGTTTATCTTCAATCGTGATGTTTACGCCTTTTGTCAAAAAGGCTAATTCGCGAACACGGTTAGCAAGCAAAT CATATTCATACTCAGTTGTTTCCGTGAAGATTTCAGGATCTGGAACAAAGTGTGTAGTCGTTCCTGTATGATCCGTT TCGCCAATAACCTCAAGATCAGAAACCGGGACACCGCGGTTATAGACTTGGCGATGGATTTTTCCGTCACGGTGAAC AGTCACATCAAGCTCTGTTGATAACGCGTTAAA
Sequence table
<110>Qinghai University
<120>one plants of Mo Haiwei bacillus for promoting the growth of Henry David Thoreau grass and its application
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
agagtttgat cmtggctcag 20
<210> 2
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
gcytaccttg ttacgactt 19
<210> 3
<211> 41
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
gaagtcatca tgaccgttct gcaygcnggn ggnaarttyg a 41
<210> 4
<211> 44
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
agcagggtac ggatgtgcga gccrtcnacr tcngcrtcng tcat 44
<210> 5
<211> 1407
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 5
cagatgggag cttgctccct gatgttagcg gcggacgggt gagtaacacg tgggtaacct 60
gcctgtaaga ctgggataac tccgggaaac cggggctaat accggatgct tgtttgaacc 120
gcatggttca aacataaaag gtggcttcgg ctaccactta cagatggacc cgcggcgcat 180
tagctagttg gtgaggtaac ggctcaccaa ggcaacgatg cgtagccgac ctgagagggt 240
gatcggccac actgggactg agacacggcc cagactccta cgggaggcag cagtagggaa 300
tcttccgcaa tggacgaaag tctgacggag caacgccgcg tgagtgatga aggttttcgg 360
atcgtaaagc tctgttgtta gggaagaaca agtaccgttc gaatagggcg gtaccttgac 420
ggtacctaac cagaaagcca cggctaacta cgtgccagca gccgcggtaa tacgtaggtg 480
gcaagcgttg tccggaatta ttgggcgtaa agggctcgca ggcggttcct taagtctgat 540
gtgaaagccc ccggctcaac cggggagggt cattggaaac tggggaactt gagtgcagaa 600
gaggagagtg gaattccacg tgtagcggtg aaatgcgtag agatgtggag gaacaccagt 660
ggcgaaggcg actctctggt ctgtaactga cgctgaggag cgaaagcgtg gggagcgaac 720
aggattagat accctggtag tccacgccgt aaacgatgag tgctaagtgt tagggggttt 780
ccgcccctta gtgctgcagc taacgcatta agcactccgc ctggggagta cggtcgcaag 840
actgaaactc aaaggaattg acgggggccc gcacaagcgg tggagcatgt ggtttaattc 900
gaagcaacgc gaagaacctt accaggtctt gacatcctct gacaatccta gagataggac 960
gtccccttcg ggggcagagt gacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga 1020
tgttgggtta agtcccgcaa cgagcgcaac ccttgatctt agttgccagc attcagttgg 1080
gcactctaag gtgactgccg gtgacaaacc ggaggaaggt ggggatgacg tcaaatcatc 1140
atgcccctta tgacctgggc tacacacgtg ctacaatgga cagaacaaag ggcagcgaaa 1200
ccgcgaggtt aagccaatcc cacaaatctg ttctcagttc ggatcgcagt ctgcaactcg 1260
actgcgtgaa gctggaatcg ctagtaatcg cggatcagca tgccgcggtg aatacgttcc 1320
cgggccttgt acacaccgcc cgtcacacca cgagagtttg taacacccga agtcggtgag 1380
gtaaccttta tggagccagc cgccgaa 1407
<210> 6
<211> 1103
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 6
cgggcttttt ccaagttgac atcttccccg atacctgtgc cgagtgcagt aatcatagaa 60
cgaacttcgt tgttagaaag aattttatca agccttgctt tttcaacgtt taggatttta 120
cctcttagcg gcaaaatggc ttggaaatgc ctgtcacgtc cctgttttgc agatcctccg 180
gcagagtcac cctctacgat atataactcg gagatgctcg ggtctttcga agagcagtcc 240
gctaatttac cgggaaggtt tgaaatctcc aaagcgcttt tgcggcgcgt taattcacgc 300
gcttttttcg cagccattct tgctcttgct gccattaaac ctttgtcaac gatttttttt 360
gccgcatctg gattttccag cataaatgtt tccaacgccg cagaaaataa cgtatctgtg 420
atcgtccgcg cctctgagtt gcctagtttt gttttcgttt ggccttcgaa ctgcggatcc 480
gggtgtttga tcgagataat cgcggtaagc ccttctctca catcatctcc gcttaagttt 540
ggatcatttt cttttatgag tccttttttt ctggcgtaat cattgatgac acgagtcagc 600
cccgttttaa aaccggcttc gtgggtaccg ccttcgtacg tgttgatatt gtttgtaaat 660
gagtaaatat tgcttgtgta gctgtcattg tattgcagag cgacttcaac cgtaatgccg 720
tccttttcgc cttcaatata aatcggctct tcatggacaa cttctttgga gcggtttaaa 780
tactctacat agcttttaat tccgccttcg taatgatact catttttgcg ctcttggcct 840
tcacgtttat cttcaatcgt gatgtttacg ccttttgtca aaaaggctaa ttcgcgaaca 900
cggttagcaa gcaaatcata ttcatactca gttgtttccg tgaagatttc aggatctgga 960
acaaagtgtg tagtcgttcc tgtatgatcc gtttcgccaa taacctcaag atcagaaacc 1020
gggacaccgc ggttatagac ttggcgatgg atttttccgt cacggtgaac agtcacatca 1080
agctctgttg ataacgcgtt aaa 1103

Claims (6)

1. the Mo Haiwei bacillus of one plant of rush herbage growth, the strain classification are named as Mo Haiwei bacillus (Bacillus Mojavensis), it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 11st, 2018, Culture presevation number is CGMCC NO.15579, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
2. a kind of microbial inoculum for promoting herbage growth, which is characterized in that include Mo Haiwei gemma described in claim 1 in the microbial inoculum Bacillus.
3. a kind of microbial-bacterial fertilizer for promoting herbage growth, which is characterized in that include described in claim 1 in the microbial-bacterial fertilizer Mo Haiwei bacillus.
4. Mo Haiwei bacillus described in claim 1 is preparing the application in the microbial-bacterial fertilizer for promoting herbage growth.
5. described in Mo Haiwei bacillus described in claim 1, microbial inoculum as stated in claim 2 or claim 3 Microbial-bacterial fertilizer promote herbage growth, the application in lignocellulose degradation or antagonism Fungal Forage Diseases.
6. a kind of method for promoting herbage growth, which is characterized in that be at least one of following (1)~(3):
(1) seed soaking is carried out to grass seed using the fermentation liquid of bacterial strain described in claim 1；
(2) root irrigation is carried out to herbage seedling using the fermentation liquid of bacterial strain described in claim 1；
(3) it is applied fertilizer using microbial-bacterial fertilizer as claimed in claim 3 to herbage.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110484463A (en) * 2019-07-16 2019-11-22 中国科学院微生物研究所 One plant of Mo Haiwei bacillus B282 and its application
CN110628661A (en) * 2019-07-05 2019-12-31 哈尔滨师范大学 Bacillus mojavensis CJX-61 and application thereof
CN114369550A (en) * 2022-01-04 2022-04-19 青海大学 Bacillus amyloliquefaciens for promoting oat growth and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103931659A (en) * 2014-04-30 2014-07-23 武汉虹睿生物科技开发有限公司 Application of bacillus mojavensis KJS-3 as biopesticide
CN105831160A (en) * 2015-01-16 2016-08-10 武汉虹睿生物科技开发有限公司 Composition utilizing Bacillus mojavensis KJS-3 strain or its culture to promote plant growth and prevent plant death

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103931659A (en) * 2014-04-30 2014-07-23 武汉虹睿生物科技开发有限公司 Application of bacillus mojavensis KJS-3 as biopesticide
CN105831160A (en) * 2015-01-16 2016-08-10 武汉虹睿生物科技开发有限公司 Composition utilizing Bacillus mojavensis KJS-3 strain or its culture to promote plant growth and prevent plant death

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BACON, CW等: "Potential for control of seedling blight of wheat caused by Fusarium graminearum and related species using the bacterial endophyte Bacillus mojavensis", 《BIOCONTROL SCIENCE AND TECHNOLOGY》 *
谢永丽: "青海不同生境芽孢杆菌多样性分析、生防菌筛选及其抗病促生机理研究", 《中国博士学位论文全文数据库 农业科技辑》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110628661A (en) * 2019-07-05 2019-12-31 哈尔滨师范大学 Bacillus mojavensis CJX-61 and application thereof
CN110484463A (en) * 2019-07-16 2019-11-22 中国科学院微生物研究所 One plant of Mo Haiwei bacillus B282 and its application
CN110484463B (en) * 2019-07-16 2021-04-27 中国科学院微生物研究所 Bacillus mojavensis B282 and application thereof
CN114369550A (en) * 2022-01-04 2022-04-19 青海大学 Bacillus amyloliquefaciens for promoting oat growth and application thereof
CN114369550B (en) * 2022-01-04 2023-07-25 青海大学 Bacillus amyloliquefaciens for promoting oat growth and application thereof

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