CN107099468A - One plant of red bacillus and its application - Google Patents
One plant of red bacillus and its application Download PDFInfo
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- CN107099468A CN107099468A CN201710165834.1A CN201710165834A CN107099468A CN 107099468 A CN107099468 A CN 107099468A CN 201710165834 A CN201710165834 A CN 201710165834A CN 107099468 A CN107099468 A CN 107099468A
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- salt
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- red bacillus
- cucumber
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- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 17
- 150000003839 salts Chemical class 0.000 claims abstract description 11
- 241000598149 Roseobacter sp. Species 0.000 claims abstract description 8
- 230000008635 plant growth Effects 0.000 claims abstract description 8
- 244000005700 microbiome Species 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 238000009629 microbiological culture Methods 0.000 claims 1
- 239000002689 soil Substances 0.000 abstract description 22
- 230000001737 promoting effect Effects 0.000 abstract description 7
- 230000012010 growth Effects 0.000 abstract description 6
- 230000015784 hyperosmotic salinity response Effects 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 230000009466 transformation Effects 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 20
- 240000008067 Cucumis sativus Species 0.000 description 18
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 17
- 241000894006 Bacteria Species 0.000 description 15
- 241000196324 Embryophyta Species 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 241000209140 Triticum Species 0.000 description 5
- 235000021307 Triticum Nutrition 0.000 description 5
- 230000035784 germination Effects 0.000 description 4
- 239000013535 sea water Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 239000005955 Ferric phosphate Substances 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 244000305550 Streptopus amplexifolius Species 0.000 description 1
- 235000001231 Streptopus amplexifolius Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229940032958 ferric phosphate Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- WBJZTOZJJYAKHQ-UHFFFAOYSA-K iron(3+) phosphate Chemical compound [Fe+3].[O-]P([O-])([O-])=O WBJZTOZJJYAKHQ-UHFFFAOYSA-K 0.000 description 1
- 229910000399 iron(III) phosphate Inorganic materials 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000007483 microbial process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 235000019354 vermiculite Nutrition 0.000 description 1
- 239000010455 vermiculite Substances 0.000 description 1
- 229910052902 vermiculite Inorganic materials 0.000 description 1
Classifications
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- 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
-
- 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
-
- 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
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Materials Engineering (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
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Abstract
The present invention provides the red bacillus that one plant of preserving number is CGMCC No.13352（Roseobacter sp.）, the strain number is red bacillus 32, belongs to biological technical field.Present invention also offers application of the red bacillus 32 that preserving number is CGMCC No.13352 in terms of promoting plant growth and improving plant salt endurance.The preserving number that the present invention is provided has the obvious effect for promoting plant growth and improving salt tolerance for CGMCC No.13352 red bacillus 32, it can be widely applied to the transformation in salt-soda soil and promote the growth of salt-soda soil crops, with good ecology and social benefit.
Description
Technical field
The present invention relates to one plant of red bacillus（Roseobacter sp.）And the application of the bacterial strain；More precisely, being related to
A kind of preserving number is CGMCC No.13352 red bacillus（Roseobacter sp.）, further relate to the red bacillus
（Roseobacter sp.）Application in terms of promoting plant growth and improving plant salt endurance.
Background technology
Salt-soda soil be it is a series of acted on by salt and alkali component in the soil body, including various solonchak, alkaline earth and other are different degrees of
The general name of all kinds soil of salinization and alkalization.The area in global salt-soda soil is 9.5 hundred million hectares, the current various saline-alkali soil of China
Area is 99,130,000 hectares.Arable area can not only be expanded, alleviate crisis in food by reasonably developing salt-soda soil, and
It can also be improved the ecological environment by planting plants, improve the quality of life of people.
However, because the salinity containing high level causes that plant growth is difficult, yield poorly down in salt-soda soil, serious salt
Alkali soil region plant can hardly survive.The screening of salt tolerant growth promoting microorganism is carried out, and applied to the improvement in salt-soda soil, having can
The recycling mode of low cost be able to can be provided for alkaline land improving.
The content of the invention
Present invention offer is a kind of can to improve salt resistance of plants and the red bacillus strain with growth promoting effect.
One plant of red bacillus strain, depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center, protects
The Tibetan date is on November 28th, 2016, preserving number：CGMCC No.13352.
Bacterial strain of the present invention is isolated from China's Sea Water of The Bohai Gulf sample, with the effect for promoting plant growth,
Crops can be effectively promoted to germinate and grow in the environment for have salt stress, improve the Soil Micro-environment in salt-soda soil, relatively
In chemical modifying mode, it is possible to reduce the destruction to Soil Micro-environment, contribute to the sustainable use to salt-soda soil, be salt-soda soil
Utilization provide it is a kind of effectively, environmental protection, safety microbial process.The present invention can be to carry out salt-soda soil using microbial resources
Improvement and the exploitation of promotion salt-soda soil provide strain resource and utilize foundation.
Biological sample preservation information
Red bacillus 3-2, taxology is named asRoseobacter Sp., it has been preserved in Chinese microorganism strain preservation conservator
Can common micro-organisms center, abbreviation CGMCC, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences microorganism
Research institute, postcode 100101, culture presevation number is 13352.
Embodiment
The separation of the marine bacteria of embodiment 1
53, the surface seawater sample in China Bohai Sea Gulf Cangzhou marine site is taken using surface water sampling device, using 2216E culture mediums
（It is formulated and is：Peptone 5g, dusty yeast 1g, ferric phosphate 0.1g, agar powder 15g, seawater 1000ml, adjust pH to be 7.6,121 with NaOH
DEG C sterilizing 20min after use）, the separation of the method progress Oceanic Samples of flat board is applied using dilution.By the flat board of coating at 28 DEG C
Cultivated in incubator, after flat board grows bacterium colony, the single bacterium colony of picking different shape is pure using line partition method 3 separation of progress
Change, obtain the pure culture bacterium colony of marine bacteria, 126 plants of marine bacteria is obtained altogether.By the liquid culture of single bacterium colony with 60% it is sweet
Oil presses 1:After 1 ratio is mixed, it is stored in -80 DEG C of ultra low temperature freezers.
The screening of the ocean Promoting bacteria of embodiment 2
The bacterial strain of ocean that is isolated from of preservation is activated on LB flat boards, afterwards by the marine bacteria activated by 1% inoculum concentration
It is inoculated into the triangular flask equipped with 20ml sea water mediums, 28 DEG C of culture 12h.Bacterium solution 5000rpm is centrifuged into 10min, outwelled
Clear liquid simultaneously adds the concussion of 5ml sterilized waters, and thalline is resuspended.Then 5000rpm centrifuges 10min again, outwells supernatant.With sterile
Water is resuspended thalline and adjusts OD600For 0.2-0.3, cucumber, wheat seed are soaked after 4h in the bacterium solution for mix up concentration, by seed
The neat sterilized water for being placed on sterilized filter paper and adding 5ml.It is put into 25 ± 2 DEG C of plant culturing room to cultivate 3 days, timing
Measure bud length and root is long.Table 1 be part bacterial strain to cucumber, the promotion result of wheat germination, wherein bacterial strain 3-2 is to cucumber and wheat
Germination, the growth of root length, bud length have good facilitation, and compared with the control, promotions of the 3-2 to cucumber root length reaches
21.9%, the promotion rate to wheat root length and bud length is respectively 32.0% and 16.8%.
The part bacterial strain of table 1 is to cucumber, the facilitation effect of wheat germination
。
Biological controls of the 3-2 of embodiment 3 to earth culture cucumber
By vermiculite and Nutrition Soil 1:1 is well mixed, and is added in square flowerpot.The square flowerpot for filling in soil is put into two big
In white pallet, it is OD to be added into a pallet in 3L sterilized waters, another pallet and add 3L concentration600Be worth for 0.1 it is red
Bacillus 3-2 bacterium solution.After after soil completely moistening, at least two cucumber seeds are cultivated to square flowerpot, culture is put into
Cultivate room.When growing a true leaf Deng cucumber, the consistent cucumber seedling of one plant of growing way is stayed per basin.After hot-house culture 6 weeks, by cucumber
Plant, which is taken out, to be cleaned, and measures the data such as its plant height, dry weight and weight in wet base（Table 3）.According to table 2 it can be seen that adding bacterial strain 3-2 place
Manage its plant height, dry weight, weight in wet base and be all higher than blank control, illustrate that bacterial strain 3-2 administrations in soil can be obviously promoted cucumber growth.
Table 2 applies influences of the bacterial strain 3-2 to cucumber growth
。
Facilitations of the 3-2 of embodiment 4 to salt tolerance of cucumber
Bacterial strain 3-2 is cultivated as described in Example 2.The consistent cucumber seeds of germination are planted to containing different water
In the water planting box for training liquid.Experiment sets 4 groups of processing altogether, is blank control respectively（For running water）, bacterium processing（Contain in running water
Concentration is 1 × 104Cfu/mL bacterial strains 3-2）, salt treatment（Sodium chloride concentration is 50mmol/L polymer solution in water）At salt plus bacterium
Reason（It is 1 × 10 to have concentration in water4Cfu/mL bacterial strains 3-2 and 50mmol/L sodium chloride）.4 groups of processing for having planted cucumber are put
Enter 25 ± 2 DEG C of temperature, light dark period for 12h:Cultivated in 12h plant culturing room, the life of cucumber is observed in culture after 20 days
Long situation, and measure root length, stem length and length of blade data.As a result as shown in table 3, cucumber in bacterial strain 3-2 treatment group is added
The root length of seedling, stem are high and length of blade is all higher than blank control, show that bacterial strain can also promote plant growth in water planting environment.
Compare the growing state for adding 3-2 bacterial strains in saline environment and being not added with the cucumber of 3-2 bacterium, it was also found that addition 3-2 bacterial strains can
Alleviate harm of the salt stress to plant growth, show that bacterial strain 3-2 can improve the salt tolerance of plant.
The water planting cucumber of table 3 and salt tolerance experimental result
。
The growth-promoting salt-enduring strain 3-2 of embodiment 5 identification
16SrDNA is identified：Bacterial strain 3-2 is inoculated into ocean culture medium, 28 DEG C of h of shaking table culture 24, using Tiangeng biochemical technology
（Beijing）Co., Ltd's genome DNA extracting reagent kit extracts the genomic DNA of the bacterial strain, then using the STb gene of extraction as
Template, with 27f（5′-AGAGTTTGATCCTGGCTCAG -3′）With 1492r (5 '-TACGGCTACCTTGTTACGAC TT -3
') primer pair enter performing PCR amplification.PCR reaction systems are 50 μ l, and reaction system is：Genomic DNA 1 μ L, 10 × PCR
5 μ L, 27f primers of buffer, 1 μ L, 1492r primer, 14 μ L, Taq enzyme (5U/ μ L) of μ L, dNTPs 1 μ L, ddH2O 36μL.Instead
The condition is answered to be：94 DEG C of min of pre-degeneration 5；94 DEG C of denaturation 1min, 50 DEG C of annealing 1min, 72 DEG C of extension 2min, are circulated 35 times；72
DEG C extension 10 min.PCR primer carries out electrophoresis detection with 1% Ago-Gel, the PCR for being about 1500bp by PCR stripe sizes
Product is served after the sequencing of Hai Sheng works Bioisystech Co., Ltd, by the DNA sequence dna of acquisition（See Seq1）, GenBank is inputted, is used
Blast programs are compared analysis with all sequences in database.The structure of phylogenetic tree is carried out using MEGA4.1.Through
Cross sequence analysis to show, the bacterial strain belongs to one kind of red Bacteriaceae.
Claims (3)
1. one plant of red bacillus（Roseobacter sp.）, it is characterised in that：The red bacillus（Roseobacter sp.）In 2016
On November 28, in has carried out preservation, preservation in China Committee for Culture Collection of Microorganisms's General Microbiological Culture collection
Number be CGMCC No. 13352.
2. described in claim 1 red bacillus (Roseobacter sp.）Applications of the 3-2 in plant growth is promoted.
3. described in claim 1 red bacillus (Roseobacter sp.）Applications of the 3-2 in plant salt endurance is improved.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710165834.1A CN107099468B (en) | 2017-03-20 | 2017-03-20 | Rhodobacter and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710165834.1A CN107099468B (en) | 2017-03-20 | 2017-03-20 | Rhodobacter and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107099468A true CN107099468A (en) | 2017-08-29 |
| CN107099468B CN107099468B (en) | 2020-08-21 |
Family
ID=59675517
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710165834.1A Expired - Fee Related CN107099468B (en) | 2017-03-20 | 2017-03-20 | Rhodobacter and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107099468B (en) |
-
2017
- 2017-03-20 CN CN201710165834.1A patent/CN107099468B/en not_active Expired - Fee Related
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| Publication number | Publication date |
|---|---|
| CN107099468B (en) | 2020-08-21 |
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