CN107099477A - One plant of plant endogenesis Bacillus flexus and its application with salt resistance ability - Google Patents
One plant of plant endogenesis Bacillus flexus and its application with salt resistance ability Download PDFInfo
- Publication number
- CN107099477A CN107099477A CN201710416367.5A CN201710416367A CN107099477A CN 107099477 A CN107099477 A CN 107099477A CN 201710416367 A CN201710416367 A CN 201710416367A CN 107099477 A CN107099477 A CN 107099477A
- Authority
- CN
- China
- Prior art keywords
- plant
- salt
- growth
- bacillus flexus
- resistance ability
- 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
Classifications
-
- 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
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Environmental Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Botany (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses one plant of plant endogenesis Bacillus flexus (Bacillus flexus) PE4941 with salt resistance ability, is preserved in Guangdong Culture Collection on May 10th, 2017, and deposit number is GDMCC 60176；And its for promoting the application in plant growth under salt stress.The naringi crenulata endogenetic bacteria Bacillus flexus PE4941 of separation screening of the present invention have fixed nitrogen, produce a variety of growth-promoting features such as siderophore, and it is resistant to 8% NaCl, pass through pouring root tieback naringi crenulata seedling, with significantly slowed under salt stress salt damage symptom, increase biomass so as to promote its grow effect, the salt tolerance of plant is favorably improved, the use of chemical fertilizer is reduced.
Description
Technical field
The invention belongs to microbial manure and crops adverse circumstance Protective strategy field, it is related to plants endogenetic bending gemma
Bacillus and its application.
Background technology
The salinization of soil is one of main abiotic stress factor of the production of current threat Global Agriculture and crop yield,
Irrigating land for the whole world nearly 50% is all threatened by different degrees of salt stress at present, China about 9.9 × 107hm2Soil
There are problems that salt marsh harm, and compromised area increases year by year.How to improve and using the salinization soil of large area,
And it is current urgently to be resolved hurrily with life science in the world to protect and improve important economical crops resistance salt stress
Scientific And Technical Problems.Current primary biological modification method measure is the crop for cultivating and planting Salt And Alkali Tolerance, such as planting beet, jade
Rice, soybean, and salt-tolerant plant alkali paulin, Chinese tamarisk etc., in addition with the research of salt tolerant genetically modified plants, but such method is often
It is cycle length, costly, and plant transgenic method does not also accept extensively accreditation by society.Endophytic bacterium refers to exist
In health plant organization internal, a class new microbe resource of plant disease is not caused.The quasi-microorganism is colonized in plant,
The influence of external environment is not easily susceptible to, research finds that many endogenetic bacterias have fixed nitrogen, Soluble phosphorus, produce auxin, iron
Carrier, the function of producing antibiotic and inducing plant resistance, endophytic bacterium are promoting plant growth and environment stress guarantor
Effect potentiality in terms of shield have been subjected to the concern of more and more researchers.
Halophytes naringi crenulata is lumbaginaceae medicinal herb plant, and the category is described as " pionner " of alkaline land improvement,
It is that China passes with the multiple efficacies such as clearing heat and detoxicating, anti-aging and anticancer simultaneously containing effective ingredients such as a variety of flavones and polysaccharide
One of system Chinese medicine, clinical medical value is big.But wild naringi crenulata quantity die-offs at present, it would be highly desirable to protect.It is relevant at present beneficial thin
Bacterium promotes also to be rarely reported in terms of improving its anti-adversity ability under wild naringi crenulata growth and high-salt stress.Microorganism has volume
It is small, grow the advantage such as fast, easy to control, therefore excavate beneficial microbe microorganism resource out of salt-soda soil grows naringi crenulata body, it is right
In promoting, the growth and adverse circumstance protection of Wild Medicinal herbaceous plant naringi crenulata are significant.
The content of the invention
The purpose of the present invention is that there is provided a kind of plant endogenesis bending gemma with salt resistance ability for above-mentioned technical problem
Bacillus.
It is a further object of the present invention to provide the plant growth-promoting agent of endogenetic bacteria production.
Promote under salt stress the application in plant growth it is yet another object of the invention to provide the endogenetic bacteria.
The purpose of the present invention can be achieved through the following technical solutions：
The present invention provides one plant of plant endogenesis Bacillus flexus (Bacillus flexus) with salt resistance ability
PE4941, is preserved in Guangdong Culture Collection on May 10th, 2017, and deposit number is GDMCC 60176.
Plant endogenesis Bacillus flexus (Bacillus flexus) PE4941 of described salt resistance ability, its feature exists
In the 16S rRNA gene orders of the strain are as shown in SEQ ID NO.1.
It is a kind of to contain plant salt tolerance growth-promoting agent of the described preserving number for GDMCC 60176 PE4941 bacterial strains.
The plant endogenesis Bacillus flexus of described salt resistance ability promotes the application in plant growth under salt stress, its
Described in plant be limonium sinenseKuntze.
Contain plant salt tolerance growth-promoting agent of the described preserving number for GDMCC 60176 PE4941 bacterial strains.It is by following
Method is produced：
1) by the Bacillus flexus PE4941 that deposit number is GDMCC 60176,250ml LB Liquid Cultures are inoculated in
In base (10g tryptones, 5g dusty yeasts, 30g NaCl, pH 7.2,1000ml water)；
2) 37 DEG C, after 150rpm Shaking cultures 48h, bacterium is collected by centrifugation in 8000rpm when thalli growth to exponential phase
Body；
3) after using sterilized water washing thalline twice, the OD values of sterilized water regulation thalline are 0.5-0.6, and cell concentration is 107-
108Cfu/mL, is described salt tolerant growth-promoting agent.
The plant salt tolerance growth-promoting agent of the described bacterial strain containing PE4941 promotes the application in plant growth under salt stress.
The plant salt tolerance growth-promoting agent of the described bacterial strain containing PE4941 promotes answering in limonium sinenseKuntze growth under salt stress
With.
The plant salt tolerance growth-promoting agent of the described bacterial strain containing PE4941 is used for the method for the Plant growth-promoting effect of salt stress, its feature
It is, the bacterium solution (50ml) for being 0.5-0.6 with the OD values of thalline containing PE4941 carries out seedling root irrigation, pouring root once, connects weekly
Continue pouring root four times.
Detailed description of the invention：
The present invention passes through to the separation of medicinal herb plant limonium sinenseKuntze endogenetic bacteria and salt tolerant and the sieve of growth promotion feature
Choosing, obtains the bacterial strain of one plant stronger salt tolerant growth-promoting activity, numbering is PE4941.Pass through morphological feature, Physiology and biochemistry and 16S
RRNA gene sequencing and Phylogenetic Analysis, it is determined that its taxonomy.Verified by potted plant experiment, it is interior that the present invention is provided
Endophytic bacteria Bacillus flexus PE4941 be it is a kind of have promote limonium sinenseKuntze growth, significantly improve under its salt stress
The method of adaptability, can ferment for bacterial manure, improve plant salt tolerance ability.
Isolated strains are Bacillus flexus, and code name is PE4941, and the bacterial strain is on May 10th, 2017 in the micro- life in Guangdong Province
Preservation is registered by thing DSMZ, and deposit number is GDMCC 60176.
It is specific such as SEQ ID simultaneously the invention provides the 16S rRNA gene orders (1400bp) of the Bacillus flexus
Shown in NO.1.
In addition, the invention provides Bacillus flexus PE4941 bacteria suspension preparation method, concretely comprising the following steps：Picking exists
24-48h Bacillus flexus PE4941 single bacterium colonies are grown on LB solid plates, are inoculated in 250ml LB fluid nutrient mediums
(10g tryptones, 5g dusty yeasts, 30g NaCl, pH 7.2,1000ml water), after 150rpm Shaking cultures 48h, treats bacterium by 37 DEG C
Thalline is collected by centrifugation in 8000rpm when body grows to exponential phase, with sterilized water washing thalline twice after, sterilized water regulation thalline
OD values be 0.5-0.6, cell concentration is 107-108cfu/mL。
Simultaneously there is provided the fertilizing method of above-mentioned bacteria suspension, comprise the following steps：Use Nutrition Soil with vermiculite with volume ratio
4:1 is mixed to get potting media, is then sterilized one hour in 121 DEG C, dispenses into potted flower pot that (9cm length × 9cm is wide after cooling
× 9cm is high), per basin about 200g.Full grains, grass seed of the same size of enriching blood are chosen, through surface sterilization (75% ethanol 15
Second, 0.1%HgCl 6-7 minutes), put into and nursery is carried out in sterile soil, growth is chosen after germination grows two panels true leaf
Consistent seedling carries out pot experiment, per one seedling of basin, and seedling is in (25 DEG C of greenhouse-grown；12h illumination/12h dark processings), use
OD values carry out seedling root irrigation for 0.5-0.6 bacterium solution (50ml), weekly pouring root once, with 200mM's after continuous pouring root 4 times
NaCl solution carries out salt stress processing.By pot experiment, demonstrate Bacillus flexus PE4941 under salt stress and China is mended
The growth of blood grass seedling has obvious facilitation, and plant height and fresh weight have been respectively increased 1.4 and 1.93 times.
Beneficial effect：
Compared with prior art, the naringi crenulata endogenetic bacteria Bacillus flexus PE4941 tools of separation screening of the present invention
There is fixed nitrogen, produce a variety of growth-promoting features such as siderophore, and be resistant to 8% NaCl, pass through pouring root tieback naringi crenulata seedling, tool
Have and salt damage symptom, increase biomass are significantly slowed under salt stress so as to promote the effect that it grows, be favorably improved the resistance to of plant
Salt, reduces the use of chemical fertilizer.
Brief description of the drawings
Fig. 1 bacterial strains PE4941 cultivate the bacterium colony photo of 48 hours on LB flat boards
Fig. 2 are based on bacterial strain PE4941 phylogenetic trees gene constructed 16S rRNA
Fig. 3 connect bacterium and do not connect the comparison of bacterium naringi crenulata seedling growth conditions under the conditions of high-salt stress
Fig. 4 high salts and without the comparison that bacterium is connect under condition of salt stress Yu bacterium naringi crenulata seedling fresh weight is not connect
Fig. 5 high salts and without the comparison that bacterium is connect under condition of salt stress Yu bacterium naringi crenulata Seedling Height is not connect
Biomaterial preservation information
The plant endogenesis Bacillus flexus PE4941 of salt resistance ability, Classification And Nomenclature is Bacillus flexus (Bacillus
Flexus), Guangdong Culture Collection (GDMCC) is preserved in, address is that XianLie Middle Road, GuangZhou City, GuangDong Province is No. 100 big
No. 59 building of institute Guangdong Culture Collection, deposit number is GDMCC 60176, and preservation date is on May 10th, 2017.
Specific embodiment
Embodiment 1：
1. endophyte is separated
Plant sample is gathered from the salt-soda soil of tidal flat of Jiangsu Province, and herborization takes back laboratory after completion in 48 hours
Strain isolation.Sample is first rinsed well with running water, removes the soil and attachment on surface, plant sample then is cut into 2cm
× 2cm sizes, soak sample 3-5min with the sodium hypochlorite of 5% concentration in aseptic superclean bench, then add sterile
2.5% sodium thiosulfate processing 10min, with the plant 5 times of sterile water wash surface sterilization, the alcohol-pickled sterilization for plus 75%
3min, sterile water wash sample 3-5 times, then with sterile mortar plant is fully ground broken, plus a certain amount of sterilized water and it is diluted to
10-1-10-4Concentration gradient, takes 10-2-10-4The Sample dilution of concentration is coated on LB isolation mediums (10g tryptones, 5g
Dusty yeast, 30g NaCl, pH 7.2, agar 18g, 1000ml water), 37 DEG C of culture 2-7d, the bacterium colony grown in time be used in combination by purifying
The 4 DEG C of preservations of LB solid slopes.
2. bacterial strain is identified
Bacterial strain PE4941 form and physiological and biochemical property：Bacterial strain is in tryptone-yeast extract medium (LB), nutrition fine jade
Well-grown on a variety of culture mediums such as fat (NA), yeast extract-malt extract (ISP 2), after LB solid plates growth 48h, bacterium colony
For circular, relatively moistening, diameter about 2-3mm, near-white, opaque (Fig. 1).The NaCl that maximum concentration is 8% is resistant to, can be by
Nitrate reduction is nitrite, produces arginine hydrolase, urase and protease.
8000rpm after 37 DEG C, 150rpm Shaking cultures 48h is based on using NA Liquid Cultures, and thalline, microwave method is collected by centrifugation
Phage gene group DNA is extracted, with bacterial universal primers 27f 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492r：5'-
AAGGAGGTGATCCAGCCGCA-3' carries out the amplification of 16S rRNA gene PCRs.PCR reaction systems are：Template DNA 2 μ L, 10 ×
Buffer 5 μ L, MgCl21 μ L, 1492r (the 10 μm of ol/ of μ L, 27f (10 μm of ol/L) of (25mmol) 3 μ L, dNTP (10mmol/L) 1
L) 1 μ L, Taq enzyme (5u/ μ L) 0.5 μ L, it is limited that PCR primer delivers to raw work (Shanghai) after being detected through 0.8% agarose gel electrophoresis
Company is sequenced.Sequencing result compares sequencing result on NCBI and EzBio Cloud websites, is then entered with the softwares of Mega 6.0
Row sequence alignment and analysis, finally with adjacent method (Neighbor-Joining, NJ) constructing system chadogram and carry out system hair
Analysis is educated, sequence alignment finds the effective kind of sequence similarity highest of the bacterial strain and bacillus, with Bacillus flexus
Bacillus flexus NBRC15715TSimilitude be 100% (Fig. 2).Therefore, bacterial strain PE4941 should belong to bacillus
The member of category, is named as Bacillus flexus PE4941 (Bacillus flexus PE4941).
3. bacterial strain growth promotion performance
Fixed nitrogen potentiality is detected：The a small amount of thalline of picking be inoculated into spot planting on nitrogen-free agar (sucrose 10.0g,
CaCO31.0g, K2HPO40.5g, MgSO4·7H2O 0.2g, NaCl 0.2g, agar 15g, H2O 1000mL, pH 7.2),
It is placed in 28 DEG C of constant incubators and cultivates 7 days or so, observe whether it grows, if bacterial strain can also grow after transferring 5 times, says
The bright bacterial strain has fixed nitrogen lateral reactivity.Bacterial strain PE4941 pass on 5 times after still can on nitrogen-free agar well-grown, explanation
PE4941 has the potential of nitrogen in fixed air.
Produce ammonia Activity determination：The a small amount of strain to be tested of picking is inoculated in 5mL peptones liquid (10g/L) culture medium, three times
Repeat, in after 28 DEG C of 120r/min shaking tables shaken cultivations 3 days, 0.5mL Nessler ' s reagents are added in each test tube, if going out
Existing tan precipitate then illustrates there is production ammonia activity, if not producing ammonia without if.Chromogenic reaction is positive after bacterial strain PE4941 cultures, says
Bright PE4941 has the ability of production ammonia.
Produce siderophore Activity determination：The a small amount of thalline of picking is inoculated on CAS detection culture mediums with spot planting, is placed on 28 DEG C
Cultivated 7 days or so in constant incubator, whether observation periphery of bacterial colonies yellow halo occurs, illustrate that the bacterial strain has production iron if having
The activity of carrier.There is yellow halo after CAS plated growths in bacterial strain PE4941, illustrates that it has the ability for producing siderophore.
4. bacterial strain application test
Bacterial strain bacteria suspension preparation method：The Bacillus flexus PE4941 that picking grows 24-48h on LB solid plates is mono-
Bacterium colony, is inoculated in 250ml LB fluid nutrient mediums, 37 DEG C, after 150rpm Shaking cultures 48h, treats that thalli growth to logarithm is given birth to
Thalline is collected by centrifugation in 8000rpm when long-term, with sterilized water washing thalline twice after, the OD values of sterilized water regulation thalline are 0.5-
0.6 is used as bacteria agent.
Potted plant fertilising experiment：Use Nutrition Soil with vermiculite with volume ratio 4:1 is mixed to get potting media, then in 121 DEG C
Sterilizing one hour, is dispensed into potted flower pot (9cm length × 9cm wide × 9cm high) after cooling, often basin about 200g.Particle is chosen to satisfy
Full, grass seed of the same size of enriching blood, through surface sterilization (75% ethanol 15 seconds, 0.1%HgCl 6-7 minutes), puts sterilized soil into
Nursery is carried out in earth, the consistent seedling of growth is chosen after germination grows two panels true leaf and carries out pot experiment, per one, basin
Seedling, seedling is in (25 DEG C of greenhouse-grown；12h illumination/12h dark processings), carry out pouring root with OD values 0.5-0.6 bacterium solution (50ml)
Processing, with isometric sterile water process as control, handles weekly once, is carried out after one month with 200mM NaCl solution
Salt stress processing, control continues pouring root water, randomly selects plant and taken pictures, measured, detected within 30 days after salt stress.As a result such as
Shown in Fig. 3, the basin of first row three is that, without the limonium sinenseKuntze under salt stress, second is classified as the lower bacterium of 200mM NaCl stress from left to right
The plant of strain PE4941 processing, the 3rd is classified as the plant shoots without bacterial strain processing under 200mM NaCl stress, it can be seen that salt is coerced
It is with compeling the lower naringi crenulata growth of seedling for being inoculated with Bacillus flexus PE4941 more preferable.As shown in table 1 and Fig. 4, bending bud with the addition of
The naringi crenulata seedling of spore bacillus PE4941 microbial inoculums is after high-salt stress process, its fresh weight compared with not being inoculated with the control of endophyte
Add 1.93 times.As shown in figure 5, be inoculated with after high-salt stress process Bacillus flexus PE4941 naringi crenulata Seedling Height with
Control is compared to improving 1.4 times, while root length and the number of blade are all dramatically increased.
Table 1. is inoculated with influences of the endophyte PE4941 to limonium sinenseKuntze growth of seedling
Embodiment 2, a kind of plant salt tolerance growth-promoting agent, be prepared by the following method obtained by：
Picking grows 24-48h Bacillus flexus PE4941 single bacterium colonies on LB solid plates, is inoculated in 250ml's
In LB fluid nutrient mediums, 37 DEG C, after 150rpm Shaking cultures 48h, 8000rpm centrifugations are received when thalli growth to exponential phase
Collect thalline, with sterilized water washing thalline twice after, sterilized water regulation thalline OD values for 0.5-0.6 and obtain plant salt tolerance growth-promoting
Agent.
<110>Jiangsu Normal University
<120>One plant of plant endogenesis Bacillus flexus and its application with salt resistance ability
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1400
<212> DNA
<213>Bacillus flexus PE4941
<400> 1
cgagcgaact gattagaagc ttgcttctat gacgttagcg gcggacgggt gagtaacacg 60
tgggcaacct gcctgtaaga ctgggataac tccgggaaac cggagctaat accggataac 120
attttctctt gcataagaga aaattgaaag atggtttcgg ctatcactta cagatgggcc 180
cgcggtgcat tagctagttg gtgaggtaac ggctcaccaa ggcaacgatg catagccgac 240
ctgagagggt gatcggccac actgggactg agacacggcc cagactccta cgggaggcag 300
cagtagggaa tcttccgcaa tggacgaaag tctgacggag caacgccgcg tgagtgatga 360
aggctttcgg gtcgtaaaac tctgttgtta gggaagaaca agtacaagag taactgcttg 420
taccttgacg gtacctaacc agaaagccac ggctaactac gtgccagcag ccgcggtaat 480
acgtaggtgg caagcgttat ccggaattat tgggcgtaaa gcgcgcgcag gcggtttctt 540
aagtctgatg tgaaagccca cggctcaacc gtggagggtc attggaaact ggggaacttg 600
agtgcagaag agaaaagcgg aattccacgt gtagcggtga aatgcgtaga gatgtggagg 660
aacaccagtg gcgaaggcgg ctttttggtc tgtaactgac gctgaggcgc gaaagcgtgg 720
ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgagt gctaagtgtt 780
agagggtttc cgccctttag tgctgcagct aacgcattaa gcactccgcc tggggagtac 840
ggtcgcaaga ctgaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg 900
gtttaattcg aagcaacgcg aagaacctta ccaggtcttg acatcctctg acaactctag 960
agatagagcg ttccccttcg ggggacagag tgacaggtgg tgcatggttg tcgtcagctc 1020
gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttgatct tagttgccag 1080
catttagttg ggcactctaa ggtgactgcc ggtgacaaac cggaggaagg tggggatgac 1140
gtcaaatcat catgcccctt atgacctggg ctacacacgt gctacaatgg atggtacaaa 1200
gggctgcaag accgcgaggt caagccaatc ccataaaacc attctcagtt cggattgtag 1260
gctgcaactc gcctacatga agctggaatc gctagtaatc gcggatcagc atgccgcggt 1320
gaatacgttc ccgggccttg tacacaccgc ccgtcacacc acgagagttt gtaacacccg 1380
aagtcggtgg ggtaaccttt 1400
Claims (10)
1. one plant of plant endogenesis Bacillus flexus (Bacillus flexus) PE4941 with salt resistance ability, in May, 2017
Guangdong Culture Collection is preserved within 10th, deposit number is GDMCC 60176.
2. the plant endogenesis Bacillus flexus (Bacillus flexus) of salt resistance ability according to claim 1
PE4941, it is characterised in that the 16S rRNA gene orders of the PE4941 bacterial strains are as shown in SEQ ID NO.1.
3. the plant endogenesis Bacillus flexus (Bacillus flexus) of salt resistance ability according to claim 1
PE4941, it is characterised in that the PE4941 bacterial strains are in LB solid plate well-growns, after growth 48h, bacterium colony for it is circular,
Relatively moistening, diameter about 2-3mm, near-white are opaque.
4. a kind of preserving number containing described in claim 1 is the plant salt tolerance growth-promoting agent of GDMCC 60176 PE4941 bacterial strains.
5. the plant endogenesis Bacillus flexus PE4941 of salt resistance ability according to claim 1 promotes to plant under salt stress
Application in thing growth.
6. the plant endogenesis Bacillus flexus PE4941 of salt resistance ability according to claim 5 application, its feature exists
In described plant is limonium sinenseKuntze.
7. plant salt tolerance growth-promoting agent according to claim 4, it is characterised in that be to produce by the following method：
1) by the Bacillus flexus PE4941 that deposit number is GDMCC 60176, it is inoculated in 250ml LB fluid nutrient mediums
(10g tryptones, 5g dusty yeasts, 30g NaCl, pH 7.2,1000ml water)；
2) 37 DEG C, after 150rpm Shaking cultures 48h, thalline is collected by centrifugation in 8000rpm when thalli growth to exponential phase；
3) after using sterilized water washing thalline twice, the OD values of sterilized water regulation thalline are 0.5-0.6, and cell concentration is 107-
108Cfu/mL, is described salt tolerant growth-promoting agent.
8. the plant salt tolerance growth-promoting agent of the bacterial strain according to claim 4 containing PE4941 promotes limonium sinenseKuntze under salt stress
Application in growth.
9. the limonium sinenseKuntze that the plant salt tolerance growth-promoting agent of the bacterial strain according to claim 4 containing PE4941 is used for salt stress promotees
The method of growth, it is characterised in that the bacterium solution (50ml) for being 0.5-0.6 with the OD values of thalline containing PE4941 carries out limonium sinenseKuntze children
Seedling root irrigation, weekly pouring root once, continuous pouring root four times.
10. the plant endogenesis Bacillus flexus (Bacillus flexus) of salt resistance ability according to claim 1
PE4941, it is characterised in that the PE4941 bacterial strains can produce siderophore, fixed nitrogen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710416367.5A CN107099477B (en) | 2017-06-05 | 2017-06-05 | Endophytic curvularia lunata with salt tolerance and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710416367.5A CN107099477B (en) | 2017-06-05 | 2017-06-05 | Endophytic curvularia lunata with salt tolerance and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107099477A true CN107099477A (en) | 2017-08-29 |
| CN107099477B CN107099477B (en) | 2021-02-02 |
Family
ID=59660987
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710416367.5A Active CN107099477B (en) | 2017-06-05 | 2017-06-05 | Endophytic curvularia lunata with salt tolerance and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107099477B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136106A (en) * | 2018-09-18 | 2019-01-04 | 四川诚隆药业有限责任公司 | A kind of separation of rhizoma ligustici endophyte, breeding method and application thereof |
| CN109576177A (en) * | 2018-12-12 | 2019-04-05 | 新疆农业科学院土壤肥料与农业节水研究所(新疆维吾尔自治区新型肥料研究中心) | A kind of China microbot bacterial strain SM8 and its application in salt tolerant growth-promoting |
| CN110521510A (en) * | 2019-09-06 | 2019-12-03 | 云南农业大学 | A kind of method that paris polyphylla endogenetic bacteria promotes paris polyphylla growth of seedling |
| CN112980728A (en) * | 2021-03-08 | 2021-06-18 | 领先生物农业股份有限公司 | Bacillus flexus endophytic strain for salt-tolerant alkaline hydrolysis of silicon, phosphorus and potassium and application thereof |
| CN113337444A (en) * | 2021-06-26 | 2021-09-03 | 北京大学深圳研究院 | Bacillus flexus and application thereof in PHA (polyhydroxyalkanoate) production |
| CN114409446A (en) * | 2022-01-21 | 2022-04-29 | 领先生物农业股份有限公司 | Low-water solid-state fermentation method of drought-tolerant bacteria and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232532A (en) * | 2014-08-29 | 2014-12-24 | 北京大学工学院包头研究院 | Endophytic bacillus pumilus, and microbial agent, preparation method and application of endophytic bacillus pumilus |
| CN104263684A (en) * | 2014-09-25 | 2015-01-07 | 山东农业大学 | Siderophores-producing bacillus and applications thereof |
-
2017
- 2017-06-05 CN CN201710416367.5A patent/CN107099477B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232532A (en) * | 2014-08-29 | 2014-12-24 | 北京大学工学院包头研究院 | Endophytic bacillus pumilus, and microbial agent, preparation method and application of endophytic bacillus pumilus |
| CN104263684A (en) * | 2014-09-25 | 2015-01-07 | 山东农业大学 | Siderophores-producing bacillus and applications thereof |
Non-Patent Citations (1)
| Title |
|---|
| SHENG QIN,ET AL: "Isolation of ACC deaminase-producing habitat-adapted symbiotic bacteria associated with halophyte Limonium sinense (Girard) Kuntze and evaluating their plant growth-promoting activity under salt stress", 《PLANT SOIL》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136106A (en) * | 2018-09-18 | 2019-01-04 | 四川诚隆药业有限责任公司 | A kind of separation of rhizoma ligustici endophyte, breeding method and application thereof |
| CN109576177A (en) * | 2018-12-12 | 2019-04-05 | 新疆农业科学院土壤肥料与农业节水研究所(新疆维吾尔自治区新型肥料研究中心) | A kind of China microbot bacterial strain SM8 and its application in salt tolerant growth-promoting |
| CN109576177B (en) * | 2018-12-12 | 2022-07-12 | 新疆农业科学院土壤肥料与农业节水研究所(新疆维吾尔自治区新型肥料研究中心) | Chinese micro-rod strain SM8 and application thereof in salt tolerance and growth promotion |
| CN110521510A (en) * | 2019-09-06 | 2019-12-03 | 云南农业大学 | A kind of method that paris polyphylla endogenetic bacteria promotes paris polyphylla growth of seedling |
| CN110521510B (en) * | 2019-09-06 | 2021-05-07 | 云南农业大学 | Method for promoting growth of paris polyphylla seedlings by paris polyphylla endophytic bacteria |
| CN112980728A (en) * | 2021-03-08 | 2021-06-18 | 领先生物农业股份有限公司 | Bacillus flexus endophytic strain for salt-tolerant alkaline hydrolysis of silicon, phosphorus and potassium and application thereof |
| CN112980728B (en) * | 2021-03-08 | 2022-06-03 | 领先生物农业股份有限公司 | Bacillus flexus endophytic strain for salt-tolerant alkaline hydrolysis of silicon, phosphorus and potassium and application thereof |
| CN113337444A (en) * | 2021-06-26 | 2021-09-03 | 北京大学深圳研究院 | Bacillus flexus and application thereof in PHA (polyhydroxyalkanoate) production |
| CN114409446A (en) * | 2022-01-21 | 2022-04-29 | 领先生物农业股份有限公司 | Low-water solid-state fermentation method of drought-tolerant bacteria and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107099477B (en) | 2021-02-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102286412B (en) | Bacillus velezensis and application thereof in tomato blight antagonism | |
| CN107099477A (en) | One plant of plant endogenesis Bacillus flexus and its application with salt resistance ability | |
| CN103695356B (en) | A kind of bacillus amyloliquefaciens and microbiobacterial agent and application thereof | |
| CN101760439B (en) | Bacillus pumilus NMCC 46 and application thereof | |
| CN101880633B (en) | Fusarium proliferatum, and bacterium agent and application thereof | |
| CN105441366B (en) | Methylotrophic bacillus ZBL-1 is applied in preventing cotton verticillium wilt | |
| CN112358974B (en) | Plant endophytic fungus epicoccum nigrum FZT214 and application thereof | |
| CN104805021A (en) | Cadmium-tolerant penicillium and separation method thereof | |
| CN112342173B (en) | Bacillus belgii and application thereof | |
| CN103224904A (en) | Rhodopseudomonas strain, biocontrol microbial inoculum and biocontrol fermentation liquid, and corresponding preparation methods and application thereof in controlling phytophthora blight of pepper | |
| CN101215543A (en) | Strain for preventing and curing muskmelon bacteroidal spot disease and application thereof in field disease prevention | |
| CN104877926A (en) | Anti-Cotinus coggygria oxysporum strain and use thereof | |
| CN105238723B (en) | A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention crop verticillium wilt | |
| CN113755389B (en) | Bacillus bailii and application thereof | |
| CN105567600A (en) | Pathogen verticillium antagonistic bacterium and application thereof | |
| CN113549570B (en) | Soil antagonistic bacterium preparation of momordica grosvenori soil-borne pathogenic bacteria, microbial fertilizer and application | |
| CN112280694B (en) | Plant endophytic fungus phomopsis D2G7 and application thereof | |
| CN108841748A (en) | Sinorhizobium nitrogen-fixing bacteria strain H6 and its application | |
| CN105368739A (en) | Bacillus amyloliquefaciens strain capable of resisting fruit tree pathogenic fungi in broad spectrum and application thereof | |
| CN102191205B (en) | Bacterial strain B1 for converting insoluble phosphate into soluble phosphate | |
| CN114874953B (en) | Peanut rhizosphere biocontrol bacterium-Bacillus beleisi SW-1 and application thereof | |
| CN102154155B (en) | Brevibacillus brevis for preventing and treating plant fungus diseases and method for preparing biopesticide | |
| CN104108964A (en) | Preparation method and application of total nutrient microorganism biocontrol organic fertilizer | |
| CN103451116A (en) | Pseudomonas fluorescens Y13, and preparation method and applications thereof | |
| CN110373344A (en) | Carp streptomycete and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |