CN105754911A - Crassulaceae root growth-promoting Bacillus aryabhattai and application thereof - Google Patents

Crassulaceae root growth-promoting Bacillus aryabhattai and application thereof Download PDF

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CN105754911A
CN105754911A CN201610330179.6A CN201610330179A CN105754911A CN 105754911 A CN105754911 A CN 105754911A CN 201610330179 A CN201610330179 A CN 201610330179A CN 105754911 A CN105754911 A CN 105754911A
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crassulaceae
bacillus cereus
shi bacillus
microbial inoculum
promoting
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CN105754911B (en
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马海林
刘方春
杜秉海
马丙尧
丁延芹
李丽
杜振宇
姚良同
单连娟
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Shandong Academy of Forestry
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    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • 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

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Abstract

The invention discloses crassulaceae root growth-promoting Bacillus aryabhattai JIA2 which has been preserved in the China General Microbiological Culture Collection Center in January 7, 2016, and the preservation No. is CGMCC No. 11967. The crassulaceae root growth-promoting bacillus aryabhattai has very good specificity, can be successfully colonized in root soil of crassulaceae succulent plants, and can remarkably promote growth of the crassulaceae succulent plants.

Description

One strain Herba hylotelephii erythrosticti rhizosphere growth-promoting A Shi bacillus cereus and application thereof
Technical field
The present invention relates to a kind of Herba hylotelephii erythrosticti rhizosphere growth-promoting A Shi bacillus cereus and application thereof.
Background technology
Crassulaceae plants (Crassulaceae) is perennial meat draft, and outward appearance is small and exquisite, and plant plumpness succulence, moulding is special, incomparably graceful, is well received by consumers.Crassulaceae succulent plays an important role in afforestation and interior decoration, or the main material of roof greening.Along with the fast development of succulent industry, improving cultivation quality, shortening cultivation period, reduce cultivation cost is the urgent needs that Crassulaceae succulent produces.
The beneficial effect of plant is mainly manifested in the alimentation improving plant and the facilitation to plant growing by rhizosphere microorganism, uses a kind of method that bacterization technique improvement rhizosphere microflora is generally considered environmental friendliness, cost-effective raising plant cultivation quality and benefits.Plant growth-promoting rhizobacteria (PGPR) refers to that existence is in plant rhizosphere scope, has promotion or pathogen has the beneficial bacteria of antagonism be referred to as plant growing.In the last few years, carry out PGPR screening study and the application of many kind of plant both at home and abroad, and obtained and desirably apply effect.But about screening and the applied research of crassulaceae plants PGPR bacterial strain, have not yet to see report.
Application crassulaceae plants PGPR bacterial strain, promotes cultivation quality and the productivity effect of Crassulaceae succulent, and the sustainable development for succulent industry is significant.
Summary of the invention
The purpose of the present invention is contemplated to solve the problems referred to above, it is provided that a kind of Herba hylotelephii erythrosticti rhizosphere growth-promoting A Shi bacillus cereus and application thereof.
To achieve these goals, the present invention adopts the following technical scheme that
One strain Herba hylotelephii erythrosticti rhizosphere growth-promoting A Shi bacillus cereus (Bacillusaryabhattai) JIA2, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 7th, 2016, and deposit number is CGMCCNo.11967.
A kind of compositions, containing above-mentioned A Shi bacillus cereus.
A kind of microbial inoculum, active component is above-mentioned A Shi bacillus cereus.
Above-mentioned microbial inoculum, also includes carrier；Described carrier is solid carrier or liquid-carrier.
Preferred: the dosage form of described microbial inoculum is liquor, suspending agent, powder, granule, wettable powder or water dispersible granules.
A kind of Compound bacterium, containing above-mentioned A Shi bacillus cereus.
The application in Herba hylotelephii erythrosticti is cultivated of above-mentioned A Shi bacillus cereus, microbial inoculum or Compound bacterium.
Preferred: described crassulaceae plants is Herba Saussureae Involueratae.
Beneficial effects of the present invention:
The Herba hylotelephii erythrosticti rhizosphere growth-promoting A Shi bacillus cereus JIA2 of the present invention has very strong specific aim, it is possible to successfully surely grows in the rhizosphere soil of Crassulaceae succulent, and can remarkably promote the growth of Crassulaceae succulent.
Accompanying drawing explanation
Fig. 1 is the colonial morphology figure of bacterial strain of the present invention；
Fig. 2 is the optical microscope microscopy figure of bacterial strain of the present invention.
Detailed description of the invention
Below in conjunction with accompanying drawing, the invention will be further described with embodiment.
1. screening process
Gather 3, the Crassulaceae succulent Rhizosphere sampling sample of Crassulaceae succulent production base, Qingzhou City Huang Lou town, Shandong Province.By gradient dilution method, from Rhizosphere sampling, isolate bacteria distribution thing.By three zoning collimation methods, the antibacterial being separated to is purified, and judges bacterial strain whether purification by microscopy, the antibacterial after purification is numbered, picking list bacterium colony, is transferred on LB inclined-plane and saves backup.
To the R containing L-Trp2Connecing bacterium in A fluid medium, 28 DEG C, 180rpm, 4d cultivated by shaking table.Take 50 microlitre bacteria suspensions to drip on whiteware plate, be simultaneously introduced 50 microlitre Salkowski color solution (50ml35%HCLO4+1ml0.5mol/LFeCl3).Using add 50mg/LIAA color solution as positive control.Whiteware plate is placed in room temperature lucifuge and is observed after 30min, the color person of reddening represent can producing IAA, just sift out the antibacterial of producing IAA.
The antibacterial of the producing IAA that primary dcreening operation obtains is carried out quantitative assay, continues screening and there is the rhizosphere growth-promoting endophytic bacteria of application potential.Potted plant experiment finally by Herba hylotelephii erythrosticti a, it is determined that strain promotes the antibacterial that blue berry growth result is best, is numbered JIA2.
Described LB inclined-plane: peptone 10g；NaCl10g；Yeast extract 5g；Agar 15-20g；Water 1000ml；PH7.0-7.2,121 DEG C of sterilizing 20min.
The R of L-Trp2A fluid medium: yeast powder 0.5g；Glucose 0.5g；Tryptone 0.5g；Casamino acid 0.5g；Soluble starch 0.5g；Dipotassium hydrogen phosphate 0.3g；Magnesium sulfate heptahydrate 0.05g；Sodium Pyruvate 0.3g；It is dissolved in 1000ml water, with potassium dihydrogen phosphate, pH value is adjusted to 7.2,1L culture medium with dipotassium hydrogen phosphate and adds L-Trp 200mg)
2. the qualification of bacterial strain
(1) colony characteristics and thalli morphology
The rounded rule of bacterium colony and bacterium colony are relatively big, and rat is more moistening, slightly yellow opaque, and microscopy thalline is rod-short, has spore, as shown in Figure 1, 2.
(2) physiological and biochemical property
The Starch Hydrolysis test positive, V-P negative, mannitol test is positive, and glucose produces the acid test positive, and the arabinose test positive, lactose test is negative, and catalase test is positive.
(3) produce heteroauxing (IAA) ability to measure
JIA2 is inoculated in the R containing L-Trp2In A fluid medium, 28 DEG C, 180rpm, 4d cultivated by shaking table.By the OD600 value of spectrophotometry bacteria suspension, then by centrifugal for bacteria suspension 10000rpm 10min, take supernatant and add equal-volume Salkowski (50mL35%HClO4+1mL0.5MFeCl3) color solution, lucifuge stands 30min, measures its OD530 value.When calculating bacteria concentration OD600 value is 1, the content of IAA in unit volume fermentation liquid, the content of IAA is calculated by the standard curve of IAA gradient dilution.By measuring, it is 18.4 μ g/ (mL OD that JIA2 produces IAA content600)-1。
(4) 16SrDNA sequence analysis
16SrDNA gene order is such as shown in SEQIDNO.1.The 16SrRNA gene sequence of JIA2 is carried out BLAST comparison analysis with number Genbank according to the sequence in storehouse, result shows: JIA2 and Bacillusaryabhattai is in the similarity of a minimum branch, the 16SrRNA gene order of the nearest JIA2 of evolutionary distance and Bacillusaryabhattai (EF114313.2) together and reaches %.Comprehensive physiological and biochemical property and colony morphology characteristic, it is accredited as A Shi bacillus cereus (Bacillusaryabhattai), and carry out preservation, preservation date: on January 7th, 2016, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCCNo.11967, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postcode: 100101.
Concrete 16SrDNA sequence is as follows:
ccttagcggctagctccttacggttactccaccgacttcgggtgttacaaactctcgtgg
tgtgacgggcggtgtgtacaaggcccgggaacgtattcaccgcggcatgctgatccgcga
ttactagcgattccagcttcatgtaggcgagttgcagcctacaatccgaactgagaatgg
ttttatgggattggcttgacctcgcggtcttgcagccctttgtaccatccattgtagcac
gtgtgtagcccaggtcataaggggcatgatgatttgacgtcatccccaccttcctccggt
ttgtcaccggcagtcaccttagagtgcccaactgaatgctggcaactaagatcaagggtt
gcgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacaaccatgcacc
acctgtcactctgtcccccgaaggggaacgctctatctctagagttgtcagaggatgtca
agacctggtaaggttcttcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcg
ggcccccgtcaattcctttgagtttcagtcttgcgaccgtactccccaggcggagtgctt
aatgcgttagctgcagcactaaagggcggaaaccctctaacacttagcactcatcgttta
cggcgtggactaccagggtatctaatcctgtttgctccccacgctttcgcgcctcagcgt
cagttacagaccaaaaagccgccttcgccactggtgttcctccacatctctacgcatttc
accgctacacgtggaattccgcttttctcttctgcactcaagttccccagtttccaatga
ccctccacggttgagccgtgggctttcacatcagacttaagaaaccgcctgcgcgcgctt
tacgcccaataattccggataacgcttgccacctacgtattaccgcggctgctggcacgt
agttagccgtggctttctggttaggtaccgtcaaggtacaagcagttactcttgtacttg
ttcttccctaacaacagagttttacgacccgaaagccttcatcactcacgcggcgttgct
ccgtcagactttcgtccattgcggaagattccctactgctgcctcccgtaggagtctggg
ccgtgtctcagtcccagtgtggccgatcaccctctcaggtcggctatgcatcgttgcctt
ggtgagccgttacctcaccaactagctaatgcaccgcgggcccatctgtaagtgatagcc
gaaaccatctttcaatcatctcccatgaaggagaagatcctatccggtattagcttcggt
ttcccgaagttatcccagtcttacaggcaggttgcccacgtgttactcacccgtccgccg
ctaacgtcatagaagcaagcttctaatcagttcgctcgacttgcatgtatag
3. the A Shi bacillus cereus (Bacillusaryabhattai) the growth-promoting effect disquisition to Herba hylotelephii erythrosticti rhizosphere
The method of JIA2 three rides being saved on inclined-plane is activated at LB (peptone 10g；NaCl10g；Yeast extract 5g；Agar 15-20g；Water 1000ml；PH7.0-7.2, subpackage, 121 DEG C of sterilizing 20min) on flat board.Picking list bacterium colony is to corresponding 10ml fluid medium, and 37 DEG C, 180rpm cultivates 24h.By 1% inoculum concentration seed liquor is seeded in corresponding 50ml fluid medium and ferments, 37 DEG C, after 180rpm cultivates 2d, for the experiment of Herba hylotelephii erythrosticti.
Experiment carries out in Qingzhou, and test plant is crassulaceae plants Herba Saussureae Involueratae.Culture matrix is that 3:1 forms by the peat composed of rotten mosses and perlite mass ratio, and every box substrate is 100g.Test is totally 4 process and 1 group of comparison, often processes 12 strain Herba Saussureae Involueratae, totally 60 strain Herba Saussureae Involueratae.Herba Saussureae Involueratae Seedling is seedling, and hat width is about 1cm.Take 5mlJIA2 bacterium solution and be diluted to 150ml, uniformly spray to Herba Saussureae Involueratae cultivation matrix.Not inoculate bacterium solution (watering same amount of culture medium) as compareing, each process 10 strain, normal management.
Test Herba hylotelephii erythrosticti is taken back laboratory together with culture matrix, after measuring hat width and root system, Herba hylotelephii erythrosticti plant and root system is carefully taken off, culture matrix is loaded in valve bag at-4 DEG C and preserves.Weigh plant fresh weight after being cleaned by culture matrix on Herba hylotelephii erythrosticti plant root, and measure root system fresh weight and utilize WinRHIZO system that root system is analyzed, measure extent of the root system, surface area.Ethanol immersion is utilized to survey Herba hylotelephii erythrosticti chlorophyll content in leaf blades at laboratory.Experimental result is as shown in table 1.It can be seen that inoculation JIA2 significantly promotes the hat width of Herba Saussureae Involueratae, add plant and root system fresh weight, significantly promote the content of blade Determination of Chlorophyll a and chlorophyll b simultaneously.As can be seen here, inoculation JIA2 has very big application potential in promoting Herba hylotelephii erythrosticti plant growing.
Table 1JIA2 is effect in promoting Herba hylotelephii erythrosticti plant growing
The specific embodiment of the present invention is described in conjunction with accompanying drawing although above-mentioned; but not limiting the scope of the invention; one of ordinary skill in the art should be understood that; on the basis of technical scheme, those skilled in the art need not pay various amendments or deformation that creative work can make still within protection scope of the present invention.

Claims (8)

1. Herba hylotelephii erythrosticti rhizosphere growth-promoting A Shi bacillus cereus (Bacillusaryabhattai) JIA2, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 7th, 2016, and deposit number is CGMCCNo.11967.
2. a compositions, is characterized in that: containing the A Shi bacillus cereus described in claim 1.
3. a microbial inoculum, is characterized in that: active component is the A Shi bacillus cereus described in claim 1.
4. microbial inoculum as claimed in claim 3, is characterized in that: also include carrier, and described carrier is solid carrier or liquid-carrier.
5. microbial inoculum as claimed in claim 3, is characterized in that: dosage form is liquor, suspending agent, powder, granule, wettable powder or water dispersible granules.
6. a Compound bacterium, is characterized in that: containing the A Shi bacillus cereus described in claim 1.
7. the A Shi bacillus cereus described in claim 1, the microbial inoculum described in claim 3,4,5 or the Compound bacterium described in claim 6 application in crassulaceae plants is cultivated.
8. apply as claimed in claim 7, it is characterized in that: described crassulaceae plants is Herba Saussureae Involueratae.
CN201610330179.6A 2016-05-17 2016-05-17 One plant of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus and its application Active CN105754911B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110982743A (en) * 2019-12-16 2020-04-10 沈阳农业大学 Bacillus aryabhattai for promoting corn root development and application thereof
CN112410244A (en) * 2020-09-30 2021-02-26 山东农业大学 Bacillus aryabhattai and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103224900A (en) * 2013-04-26 2013-07-31 浙江大学 Plant endophyte and application thereof

Patent Citations (1)

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Publication number Priority date Publication date Assignee Title
CN103224900A (en) * 2013-04-26 2013-07-31 浙江大学 Plant endophyte and application thereof

Non-Patent Citations (1)

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Title
SHEETAL AMBARDAR.JYOTI VAKHLU ET AL.: "plant growth promoting bacteria from cocus sativus rhizosphere", 《WORLD J MICROBIOL BIOTECHNOL》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110982743A (en) * 2019-12-16 2020-04-10 沈阳农业大学 Bacillus aryabhattai for promoting corn root development and application thereof
CN112410244A (en) * 2020-09-30 2021-02-26 山东农业大学 Bacillus aryabhattai and application thereof
CN112410244B (en) * 2020-09-30 2022-04-05 山东农业大学 Bacillus aryabhattai and application thereof

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