CN105754911B - One plant of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus and its application - Google Patents
One plant of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus and its application Download PDFInfo
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- CN105754911B CN105754911B CN201610330179.6A CN201610330179A CN105754911B CN 105754911 B CN105754911 B CN 105754911B CN 201610330179 A CN201610330179 A CN 201610330179A CN 105754911 B CN105754911 B CN 105754911B
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Abstract
The invention discloses a kind of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus (Bacillus aryabhattai) JIA2, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on January 7th, 2016, deposit number is CGMCCNo.11967, red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus of the invention has very strong specific aim, it can successfully colonize in the rhizosphere soil of Crassulaceae succulent, and the growth of Crassulaceae succulent can be remarkably promoted.
Description
Technical field
The present invention relates to a kind of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus and its applications.
Background technique
Crassulaceae plants (Crassulaceae) are perennial meat draft, and appearance is small and exquisite, and plant plumpness succulence is made
Type is special, incomparably graceful, is well received by consumers.Crassulaceae succulent plays in afforestation and interior decoration focuses on
It acts on or the main material of roof greening.With the fast development of succulent industry, improves and cultivate quality, shorten training
It educates the period, reduce the urgent need for cultivating that cost is the production of Crassulaceae succulent.
Rhizosphere microorganism, which is mainly manifested in the beneficial effect of plant, improves the nutrient absorption of plant and to plant growth
Facilitation is generally considered a kind of environmental-friendly, economical and effective with bacterization technique improvement rhizosphere microflora
Raising plant cultivation quality and benefits method.Plant growth-promoting rhizobacteria (PGPR), which refers to, survives in plant rhizosphere range,
Have to plant growth and promotes or there is the beneficial bacteria of antagonism to be referred to as pathogen.In recent years, having carried out many both at home and abroad
The PGPR screening study and application of kind plant, and obtain ideal application effect.But about crassulaceae plants PGPR bacterial strain
Screening and application study, have not been reported yet.
Using crassulaceae plants PGPR bacterial strain, the cultivation quality and productivity effect of Crassulaceae succulent are promoted, for more
The sustainable development of meat plant industry is of great significance.
Summary of the invention
The purpose of the present invention is to solve the above-mentioned problems, provide a kind of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus and its
Using.
To achieve the goals above, the present invention adopts the following technical scheme:
One plant of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus (Bacillus aryabhattai) JIA2, January 7 in 2016
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center day, deposit number is CGMCC No.11967.
A kind of composition contains above-mentioned A Shi bacillus.
A kind of microbial inoculum, active constituent are above-mentioned A Shi bacillus.
Above-mentioned microbial inoculum further includes carrier；The carrier is solid carrier or liquid-carrier.
It is preferred that: the dosage form of the microbial inoculum is liquor, suspending agent, pulvis, granule, wettable powder or water dispersible granules.
A kind of Compound bacterium contains above-mentioned A Shi bacillus.
Application of above-mentioned A Shi bacillus, microbial inoculum or the Compound bacterium in red-spotted stonecrop cultivation.
It is preferred that: the crassulaceae plants are saussurea involucrata.
Beneficial effects of the present invention:
Red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus JIA2 of the invention has very strong specific aim, can successfully colonize in scape
The rhizosphere soil of its section succulent, and the growth of Crassulaceae succulent can be remarkably promoted.
Detailed description of the invention
Fig. 1 is the colonial morphology figure of bacterial strain of the present invention；
Fig. 2 is the optical microscopy microscopy figure of bacterial strain of the present invention.
Specific embodiment
The invention will be further described with embodiment with reference to the accompanying drawing.
1. screening process
Acquire the Crassulaceae succulent Rhizosphere sampling of the town Shandong Province's Qingzhou City Huang Lou Crassulaceae succulent production base
3, sample.By gradient dilution method, bacterium isolate is isolated from Rhizosphere sampling.By three zoning collimation methods to being separated to
Bacterium is purified, and judges whether bacterial strain purifies by microscopy, bacterium after purification is numbered, picking single colonie, is turned
It is connected on the inclined-plane LB and saves backup.
To the R containing L-Trp2Connect bacterium in A fluid nutrient medium, 28 DEG C, 180rpm, shaking table culture 4d.Take 50 microlitres of bacterium
50 microlitres of Salkowski color solution (50ml 35%HCLO are added on whiteware plate in suspension drop4+1ml 0.5mol/
L FeCl3).The color solution of 50mg/L IAA will be added as positive control.Whiteware plate is after room temperature avoid light place 30min
Observation, the color person of reddening indicate can producing IAA, just sift out the bacterium of producing IAA.
The bacterium for the producing IAA that primary dcreening operation obtains is quantitative determined, the rhizosphere growth-promoting that screening has application potential is continued
Bacterium.Finally by the potted plant experiment of red-spotted stonecrop, one plant of bacterium for promoting blueberry growth result best, number JIA2 are determined.
The inclined-plane LB: peptone 10g；NaCl 10g；Yeast extract 5g；Agar 15-20g；Water 1000ml；pH
7.0-7.2 121 DEG C of sterilizing 20min.
The R of L-Trp2A fluid nutrient medium: yeast powder 0.5g；Glucose 0.5g；Tryptone 0.5g；Casein amino
Sour 0.5g；Soluble starch 0.5g；Dipotassium hydrogen phosphate 0.3g；Epsom salt 0.05g；Sodium Pyruvate 0.3g；It is dissolved in 1000ml
PH value is adjusted to 7.2,1L culture medium with dipotassium hydrogen phosphate and potassium dihydrogen phosphate and L-Trp 200mg is added by water)
2. the identification of bacterial strain
(1) colony characteristics and thalli morphology
The rounded regular and bacterium colony of bacterium colony is larger, and surface bulge is relatively wet, yellowish opaque, and microscopy thallus is quarter butt
Shape has gemma, as shown in Figure 1, 2.
(2) physiological and biochemical property
Starch Hydrolysis test is positive, and V-P negative, mannitol test is positive, and glucose production acid test is positive, Arabic
Sugar test is positive, and lactose test is negative, and catalase test is positive.
(3) measurement of heteroauxin (IAA) ability is produced
JIA2 is inoculated in the R containing L-Trp2In A fluid nutrient medium, 28 DEG C, 180rpm, shaking table culture 4d.With point
Light photometry measures the OD600 value of bacteria suspension, and bacteria suspension 10000rpm is then centrifuged 10min, takes supernatant to be added isometric
Salkowski (50mL 35%HClO4+1mL 0.5M FeCl3) color solution, it is protected from light and stands 30min, measure its OD530 value.Meter
When calculation bacteria concentration OD600 value is 1, the content of IAA in unit volume fermentation liquid, the standard curve for passing through IAA gradient dilution is calculated
The content of IAA.By measurement, it is 18.4 μ g/ (mLOD that JIA2, which produces IAA content,600)-1。
(4) 16S rDNA sequence is analyzed
16SrDNA gene order is as shown in SEQ ID NO.1.By the 16S rRNA gene sequence of JIA2 and number Genbank evidence
Sequence in library carries out BLAST and compares analysis, the results showed that JIA2 and Bacillus aryabhattai are in a minimum together
Branch, the 16SrRNA gene order of the nearest JIA2 of evolutionary distance and Bacillus aryabhattai (EF114313.2's)
Similitude reaches %.Comprehensive physiological and biochemical property and colony morphology characteristic, are accredited as A Shi bacillus (Bacillus
Aryabhattai), and preservation is carried out, preservation date: is preserved in Chinese microorganism strain preservation management committee on January 7th, 2016
Member's meeting common micro-organisms center, deposit number are CGMCC No.11967, preservation address: BeiChen West Road, Chaoyang District, BeiJing City 1
No. 3 Institute of Microorganism, Academia Sinica of institute, postcode: 100101.
Specific 16S rDNA 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. A Shi bacillus (Bacillus aryabhattai) is to the growth-promoting effect disquisition of red-spotted stonecrop rhizosphere
The method of three rides of the JIA2 being stored on inclined-plane is activated in LB (peptone 10g；NaCl 10g；Yeast
Medicinal extract 5g；Agar 15-20g；Water 1000ml；PH 7.0-7.2, packing, 121 DEG C of sterilizing 20min) on plate.Picking single bacterium drops down onto
In corresponding 10ml fluid nutrient medium, 37 DEG C, 180rpm is cultivated for 24 hours.Seed liquor is seeded to corresponding 50ml by 1% inoculum concentration
It ferments in fluid nutrient medium, 37 DEG C, after 180rpm cultivates 2d, the experiment for red-spotted stonecrop.
Experiment carries out in Qingzhou, and test plant is crassulaceae plants saussurea involucrata.Culture substrate is by turf and perlite mass ratio
For 3:1 composition, every box matrix is 100g.Test totally 4 processing and 1 group of control, 12 plants of saussurea involucrata of every processing, totally 60 plants of saussurea involucrata.Snow
Lotus seedling is seedling, and hat width is 1cm or so.It takes 5mlJIA2 bacterium solution to be diluted to 150ml, uniformly sprays into saussurea involucrata cultivation matrix.
Not to be inoculated with bacterium solution (pouring same amount of culture medium) as control, each 10 plants of processing, normal management.
Test red-spotted stonecrop is taken back into laboratory together with culture substrate, after measuring hat width and root system, by red-spotted stonecrop plant and root system
It carefully removes, culture substrate is loaded in valve bag and is saved at -4 DEG C.After culture substrate on red-spotted stonecrop plant root is cleaned
Plant fresh weight is weighed, and measures root system fresh weight and root system is analyzed using WinRHIZO system, measures extent of the root system, surface
Product.Red-spotted stonecrop chlorophyll content in leaf blades is surveyed using ethanol immersion in laboratory.Experimental result is as shown in table 1.As can be seen that inoculation
JIA2 significantly promotes the hat width of saussurea involucrata, increases plant and root system fresh weight, while significantly promoting blade Determination of Chlorophyll a and leaf
The content of green element b.It can be seen that inoculation JIA2 has very big application potential in promoting red-spotted stonecrop plant growth.
Table 1JIA2 is acted in promoting red-spotted stonecrop plant growth
Above-mentioned, although the foregoing specific embodiments of the present invention is described with reference to the accompanying drawings, not protects model to the present invention
The limitation enclosed, those skilled in the art should understand that, based on the technical solutions of the present invention, those skilled in the art are not
Need to make the creative labor the various modifications or changes that can be made still within protection scope of the present invention.
Claims (5)
1. a kind of red-spotted stonecrop rhizosphere growth-promoting A Shi bacillus (Bacillus aryabhattai) JIA2, on January 7th, 2016
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.11967.
2. a kind of microbial inoculum, it is characterized in that: active constituent is A Shi bacillus described in claim 1.
3. microbial inoculum as claimed in claim 2, it is characterized in that: further including carrier, the carrier is solid carrier or liquid-carrier.
4. microbial inoculum as claimed in claim 2, it is characterized in that: dosage form is liquor, suspending agent, pulvis or granule.
5. A Shi bacillus described in claim 1, the described in any item microbial inoculums of claim 2-4 are cultivated in crassulaceae plants
In application, wherein the crassulaceae plants be saussurea involucrata.
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| CN103224900A (en) * | 2013-04-26 | 2013-07-31 | 浙江大学 | Plant endophyte and application thereof |
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| CN103224900A (en) * | 2013-04-26 | 2013-07-31 | 浙江大学 | Plant endophyte and application thereof |
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| Title |
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| plant growth promoting bacteria from cocus sativus rhizosphere;Sheetal Ambardar.Jyoti Vakhlu et al.;《World J Microbiol Biotechnol》;20130609;第2271-2279页 * |
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