CN103173383B - Shewanella piezotolerans 34# and application thereof to algae inhibition - Google Patents
Shewanella piezotolerans 34# and application thereof to algae inhibition Download PDFInfo
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Abstract
The invention discloses shewanella piezotolerans 34# and an application thereof to algae inhibition. The collection number of the shewanella piezotolerans 34# is CGMCC NO. 6490, and the shewanella piezotolerans 34# has the capacity of producing a long-chain QS (quorum sensing) signal and the function of inhibiting alga. A method provided by the invention has a good effect of inhibiting the red-tide algae, can be used for preventing red-tide organisms effectively, can realize the biological antagonistic prevention and control concept of controlling alga by bacteria and has field popularization value.
Description
Technical field
The present invention relates to biological technical field, more particularly, to a kind of ocean is pressure Xi Washi bacterium and its in suppression red tide algae
Application in growth.
Background technology
Red tide refer in ocean planktonic organism (espespecially algae) at short notice explosive propagation or high aggregation, cause
The abnormal ecological phenomenon (taking on a red color or brown) of water colour, is referred to as " algal tufa " or is likened to " red ghost " more.Paralic environment
Increasingly pollution and the unreasonable exploitation of marine resources, bring huge bearing pressure to ecological environment, lead to local red tide
Frequently occur, cause the ecocatastrophe of marine environment.This ecocatas-trophe causes the pollution at water source, the consumption of the energy, Yi Jishui
The reduction of dissolved oxygen in body, constitutes a serious threat to biological living environment.Part red tide plankton also produces with toxin, directly poison
Kill aquatile, bring massive losses to seashore economy and aquaculture.
China is the multiple state of red tide, and the generation of harmful algal is in ascendant trend year by year.According to National Bureau of Oceanography's statistics,
Since 20 century 70s, China's red tide occurrence frequency is increased with every 10 years 3 times of speed, at the beginning of 2000, has more than every year
30 times about fairly large red tide, calendar year 2001 occur 41,2006 93,2009 68.According to《Chinese Sea in 2010
Disaster publication》Display, there is red tide 69 times, 1.09 ten thousand square kilometres of cumulative area in coastal area of china in 2010, direct economy damages altogether
Lose 2.06 hundred million yuan.
Based on Disaster And Prevention Measures of Red Tides frequency in recent years is increasing, the trend increasingly sharpened of scope expanding day, harm, right
It is the primary premise protected the marine environment that red tide (or red tide algae) carries out effective prevention and control.Though developing in conventional research
Go out the control technology of some red tides, such as physical method, chemical method and part biological administering method, but these methods are all each deposited
In certain defect, the effect administering wawter bloom is not satisfactory.Such as yellow mud flocculence in physical method needs larger agent
Amount just can play the effect of suppression, the input of this difficulty that increased operation to a certain extent and cost；Chemical method is such as killed
Algae agent, while killing algae, also brings direct or indirect harm to other biological it is impossible to embody environmentally friendly spy
Property；Biological method, as thrown in the macro-organism (fish) of the algae that ingests, there is also and breaks food web frame, destroys biological net steady
Fixed ecological risk.
People more and more recognize in recent years, and the life of algae disappears and has inseparable relation with symbiotic microorganism,
Therefore more and more the prevention and controls of red tide are turned to microbial treatment, and this class method has increasingly been paid close attention to and recognized
Can.This field is screened and has cultivated and multiple killed algae, the microorganism of molten algae, including actinomyces, proteus, false unit cell
Bacterium, slime bacteria etc..Molten algae present in symbiotic microorganism or algal control are individual, can suppress algae by direct or indirect effect
Growth.The traveling of this function will depend on certain biomass, and the regulation and control of bacterial biomass are by colony induction signaling
(quorum sensing, QS) is adjusting.QS signal is the Language Communication mode that there is uniqueness between bacterium, and this signal can be real
Transmission (inter-transfer) and inter-species transmission (intra-transfer) in now planting.Between microorganism, QS can cause micro-
Biological a series of Physiology and biochemistry reflection, shows the sensing behavior of colony.
In algae-bacteria symbiotic system, algae-lysing bacterium can realize the Fast-propagation of itself under the regulation and control of QS, forms sociales
Group, shows competitive advantage in alga-bacterium symbiosis system, produces suppression to algal grown.Algae-lysing bacterium is as red-tide control
Potential microorganism, has caused increasing concern.It is that strict differentiation is red to the screening of algae-lysing bacterium in conventional research
Damp generation phase and non-generation phase are so that the bacterial strain lack of targeted of screening；In addition, whether have the sense of secretion colony to algae-lysing bacterium
The attributes research of induction signal was not also related to.
Content of the invention
It is an object of the present invention to provide one plant of pressure Shewanella (Shewanellapiezotolerans) 34#.
Pressure Shewanella (Shewanellapiezotolerans) 34# that the present invention provides, its preserving number is
CGMCCNO.6490.
It is a further object to provide a kind of above-mentioned pressure Shewanella (Shewanellapiezotolerans)
The application of 34# is it is characterised in that produce colony induction signaling (QS) for extracellular.
In a preferred embodiment of the present invention, extracellular product QS comprises the steps：
1) ferment pressure Shewanella (Shewanella piezotolerans) 34#, and supernatant is collected by centrifugation；
2) it is extracted with ethyl acetate described supernatant, collect organic phase, that is, obtain QS runic thing.
In said method, step 1) in, the temperature of described fermentation is 30 DEG C, and the time of described fermentation is 48h；Described centrifugation
Rotating speed be 8000r/min, time of described centrifugation is 20min, and described centrifugation radius is 13.5cm；
Step 2) in, described supernatant and ethyl acetate extraction volume ratio are 1: 2, and time of described extraction is 3h.
In said method, the temperature of described fermentation is 30 DEG C, and the time of described fermentation is 48 hours, the culture of described fermentation
Base is 2216E seawater fluid nutrient medium (formula：Peptone 5.0g, yeast extract 1.0g, ferric phosphate 0.01g, Chen Haishui 1000mL,
Adjust pH to 7.8)；
In said method, in step 2) in, also include successively removing in described organic phase after described collection organic phase
Ethyl acetate and the step of dissolving；
Ethyl acetate in the described organic phase of above-mentioned removal is specially organic phase described in rotary evaporation, collects product 1；
Above-mentioned dissolving is specially dissolves described product 1 with methyl alcohol；
In described step 1) and step 2) between also include the step that filters described supernatant；The concrete employing of above-mentioned filtration
Filter sizes are 0.22 μm.
Described pressure Shewanella (Shewanella piezotolerans) 34# being prepared by said method
CGMCCNO.6490 extracellular QS extract is also the scope of protection of the invention.
Another aspect of the present invention further relates to above-mentioned pressure Shewanella (Shewanella piezotolerans) 34# and exists
Application in preventing and treating red tide.
In above-mentioned application, described preventing and treating red tide passes through to suppress red tide algae growth to realize.
The growth of above-mentioned suppression red tide algae is embodied in following 1) -4) in 4 kinds or any of which：
1) reduce red tide algae density；
2) reduce the chlorophyll content of red tide algae；
3) reduce the photosynthetic efficiency of red tide algae；
4) change the composition of red tide algae algae border microorganism.
Above-mentioned application is to add above-mentioned pressure Shewanella 34# in the growth system of red tide algae, to suppress red tide algae
Growth；Described red tide algae is specially Alexandrium (Alexandrium sp.).
In an embodiment of the present invention, the method for concrete suppression red tide algae growth is to add in the growth system of red tide algae
Above-mentioned pressure Shewanella 34#, to suppress red tide algae to grow.
The growth system of above-mentioned red tide algae is artificial cultivation system；Above-mentioned artificial culture system is in nutrient solution by red tide algae
Middle culture, obtains artificial culture system；Nutrient solution in above-mentioned artificial culture system is f/2 nutrient solution；Above-mentioned pressure Xi Washi
Final concentration in described artificial culture system for the bacterium 34# is specially 1.0 × 104cells/ML-1.0×106cells/ML.
In said method, described red tide algae is Alexandrium (Alexandrium sp.).
Third object of the present invention is to provide a kind of red tide algae suppression biological.
The red tide algae suppression that the present invention provides is biological, is divided into above-mentioned pressure Shewanella 34#；Described red tide algae is specially
Alexandrium (Alexandrium sp.).
The experiment proves that, present invention finds a kind of pressure Shewanella
(Shewanellapiezotolerans.) 34#, this bacterium has the energy producing long-chain QS (13 carbon chain lengths, molecular weight 301)
Power；And bacterium itself has the function of algal control, can be used to suppress red tide；This bacterium has the advantage that：1) algal control screened is thin
Bacterium is derived from the marine site that red tide occurs, and uses beneficial to again rendering in natural environment；2) extracellular products demonstrates this bacterium and produces QS energy
The presence of power, is a kind of stronger bacterium of environmental suitability；3) method of algal inhibition red tide, it then follows biological " biological interpromoting relation in five elements phase
Gram " principle, secondary pollution will not be produced, be a kind of environmentally friendly method.
Brief description
Fig. 1：Various dose bacterial strain dense to Alexandrium Effects of Density；Wherein, * represents significant difference (P ＜ 0.05)；**
Represent that difference is extremely notable (P ＜ 0.01)
Fig. 2：The dense impact to Alexandrium chlorophyll content of various dose bacterium；Wherein, * represents significant difference (P ＜
0.05)；* represents that difference is extremely notable (P ＜ 0.01).
Fig. 3：The dense impact to Alexandrium photosynthetic efficiency of various dose bacterium；Wherein, * represents significant difference (P ＜
0.05)；* represents that difference is extremely notable (P ＜ 0.01).
Microorganism Deposit Information
Above-mentioned pressure Shewanella 34# bacterial strain is bacterium circle (Bacteria)；Proteus door (Proteobacteria)；
γ-deformed rod Gammaproteobacteria (Gammaproteobacterium), Alteromonas Zoopagales (Alteromonadales), Shewanella section
(Shewanellaceae), genus Shewanella (Shewanella) bacterial strain, is preserved in China Microbiological on 3rd in September in 2012
(abbreviation CGMCC, address is culture presevation administration committee common micro-organisms center：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.6490, and its Classification And Nomenclature is pressure Shewanella
(Shewanella piezotolerans)
Specific embodiment
Experimental technique used in following embodiments if no special instructions, is conventional method.
Material used, reagent etc. in following embodiments, if no special instructions, all commercially obtain.
F/2 nutrient solution Ju Ti Pei Fang is following (quality containing inorganic salts in every liter of seawater)：NaNO337.5mg,
NaH2PO42.5mg, Fe-EDTA2.5mg (FeCl31.6g+EDTA0.9g), Tyiamine Hd element 5 μ g, Biotin VH0.025 μ g,
VB120.025 μ g, CuSO4.5H2O0.0098 μ g, ZnSO4.7H2O0.022 μ g, CaCl2.6H2O0.01 μ g,
MgCl2.4H2O0.180 μ g, Na2MoO4.2H2O, 0.0063 μ g.
(Hong Kong strain, the Institute of Oceanology of the Chinese Academy of Sciences provides Alexandrium (Alexandrium catenella), produces
Product catalog number (Cat.No.)：Algae20060309)
Embodiment 1, the screening of pressure Salmonella (Shewanella piezotolerans) 34#, identification and characteristic
First, the screening of pressure Salmonella (Shewanella piezotolerans) 34#
On April 21st, 2011, marginal basins offshore (Shenzhen Shekou port) occurs local red tide, and (Shenzhen Shekou port red tide occurs
Area top layer), this time the reason red tide, algae is bent angle algae (Eucampia zoodiacusEhrenberg), belongs to Bacillariophyta
Bacillariophyceae, Fragilariaceae Fragilariaceae Schroder, extra large line Trentepohlia Thalassionema
Grunow.Fetch water sample from this region, water sample is taken back behind laboratory through filtered through gauze, removes impurities in water and large-scale particulate matter；
With after filter through 100 μm of metal grills, remove chip in water；Frustule is removed through 10 μm of membrane filtrations, filtrate is applied after
Cloth and sieve bacterium.
Above-mentioned ready filtrate is carried out doubling dilution, takes 30 μ L to coat and (join on 2216E seawater solid medium
Side：Peptone 5.0g, yeast extract 1.0g, ferric phosphate 0.01g, Chen Haishui 1000ml, agar 20g, adjust pH to 7.8), overnight train
Support, until growing clearly monoclonal.Pick out 200 plants of cultivable bacteria altogether.
Subsequently picking monoclonal (1mL) in 2216E fluid nutrient medium, shaking table is cultivated 12 hours for 30 DEG C, standby.To choose
The monoclonal bacterial strain of choosing is with 1 × 104The amount of individual/mL adds Alexandria algae culturing liquid (by Alexandrium in f/2 nutrient solution
Cultivate the nutrient solution that obtains) in as experimental group, two weeks interior suppression situations to algal grown of continuous monitoring, to be added without Dan Ke
Grand bacterial strain is control group, counts (4 × 10 times) detection algae density, each sample count 3 times, result is made even under binocular microscope
Average.Experimental result shows, adds bacterium solution can suppress algae growth after 2 days, and after 4 days, the algae cell density of experimental group declines
For the 50% of control group, during to 6 days, the algae of experimental group substantially stops growing and divides, and more than 80% algal grown is pressed down
System, algae bio amount shows pole significant difference (P ＜ 0.01) compared with control group.
It is therefore contemplated that preliminary screening obtains bacterial strain 34#.
2nd, 34#The identification of bacterial strain
1st, physiological and biochemical property identification
1.1 morphologic observation
Using 2216E solid medium, by 34 after purification#Monoclonal bacterial strain cultivates 24h at 30 DEG C, and it is blue to carry out leather
The micro- sem observation (oil mirror, 1000 times of multiplication factor) of Albert'stain Albert and thalli morphology.Colouring method reference《Microbiological Test is real
Test guidance》Method carry out (author：Gui Fang, publishing house：China Medical Science Press, publication time：In August, 2009,
ISBN：9787506742238).Test result indicate that bacterial strain is Gram-negative bacteria；Form is the corynebacteria of two ends blunt circle, tool
Flagellum, thalline size is (0.3～0.4) × (2.0～3.0) μm.
1.2 physiological and biochemical property
The detection of physiological and biochemical property carries out (Qingdao GaoKeYuan sea rich biology skill using proteus suit biochemical identification pipe
Art Co., Ltd, production code member：SHBG05).Experimental result display hydrogen sulfide is positive, and Phenylalanine dehydrogenase is negative, urase sun
Property.
2nd, Molecular Identification
Extract bacterial strain 34#DNA, using bacterial 16 S rRNA universal primer (forward primer：
AGAGTTTGATCCTGGCTCAG；Reverse primer：CTGAGCCAGGATCAAACTCT) PCR amplifies PCR primer and is
Encoding gene for 16S-rRNA, PCR primer is carried out electrophoresis detection, sends to survey by obtaining the product that stripe size is 1500bp
Sequence, this PCR primer of result (encoding gene of 16S-rRNA) has the nucleotides shown in sequence 1 in sequence table.
Proof after sequence alignment, the encoding gene of the 16S-rRNA of bacterial strain 34# and proteus door
(Proteobacteria), γ-deformed rod Gammaproteobacteria (Gammaproteobacterium), Alteromonas Zoopagales
(Alteromonadales), Shewanella section (Shewanellaceae), the 16S- of genus Shewanella (Shewanella sp.)
The encoding gene of rRNA has 99% similitude (GenBank：AJ551090).
Therefore above-mentioned bacterial strains 34#For proteus door (Proteobacteria), γ-deformed rod Gammaproteobacteria
(Gammaproteobacterium), Alteromonas Zoopagales (Alteromonadales), Shewanella section
(Shewanellaceae), genus Shewanella (Shewanella sp.) bacterial strain, was preserved in China micro- on April 7th, 2012
(abbreviation CGMCC, address is biological inoculum preservation administration committee common micro-organisms center：BeiChen West Road, Chaoyang District, BeiJing City 1
Institute 3, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCCNO.6490, and its Classification And Nomenclature is Shewanella
(Shewanella piezotolerans).
Embodiment 2：Pressure watt of Salmonella (Shewanella piezotolerans) 34# produces the detection of QS ability
1. the preparation of Shewanella.By culture 6-8 hour in 1.5 milliliters of eppendof pipe for the 34# bacterial strain of identification,
Eppendof pipe liquid amount is 0.5 milliliter, and culture medium is LB fluid nutrient medium (culture medium prescription：Tryptone 10g, yeast carries
Take thing 5g, NaCl10g, adjust pH to 7.0.Deionized water is settled to 1L.Steam sterilizing 20min under 15psi high pressure).Culture
Good Shewanella liquid detects its concentration through visible spectrophotometer, and its concentration value is about OD600=0.35.This preparation
It is standby that bacterium solution is placed in 4 DEG C of refrigerators.
2. the preparation of detection bacterial strain and detection.With reporting bacterial strain CV026 and A136 of specific detection QS to 34# bacterial strain institute
The QS producing is detected, wherein CV026 reporting bacterial strain is applied to detection short chain QS (4-6 carbochain), and A136 is applied to detection length
Chain QS (6-14 carbochain).The detection method of short chain QS is as follows：First CV026 bacterial strain is added LB semisolid culturemedium (culture basigamy
Side：Tryptone 10g, yeast extract 5g, NaCl10g, 0.5 gram of agar, adjust pH to 7.0.Deionized water is settled to 1L) system
Become final concentration of 1 × 106The flat board of individual/ML；Subsequently a diameter of 4 millimeters of white filter paper is placed on flat board；Finally in filter
34# bacterial strain (point sample amount be 2ul) is put on the scraps of paper, cultivates more than 12 hours in 30 DEG C, according to purple chromosphere is had or not on filter paper
Exist and judge whether 34# bacterial strain has short chain QS production capacity.Purple chromosphere is had to indicate short chain QS production capacity, no purple color
Circle then represents do not there is short chain QS production capacity.From the point of view of the result of this patent of invention, result is feminine gender, has no that colour developing is anti-
Should, therefore tentatively conclude that 34# bacterial strain does not have the production capacity of short chain QS.
The detection method of long-chain QS is as follows：First A136 bacterial strain is added LB semisolid culturemedium (culture medium prescription：Tryptose
Peptone 10g, yeast extract 5g, NaCl10g, 0.5 gram of agar, adjust pH to 7.0.Deionized water is settled to 1L) make final concentration
For 1 × 106The flat board of individual/ML；Subsequently a diameter of 4 millimeters of white filter paper is placed on flat board；Again by 2ul20mg/ml's
X-gal point is added on filter paper；Last 34# bacterial strain (point sample amount is 2ul) is put on filter paper, in 30 DEG C cultivate 12 hours with
On, judge whether 34# bacterial strain has long-chain QS production capacity according to blue chromosphere presence is had or not on filter paper.There is blue chromosphere table
It is shown with long-chain QS production capacity, no blue chromosphere then represents do not there is long-chain QS production capacity.From the point of view of experimental result, result
Display is positive, assumes chromogenic reaction, therefore concludes that 34# bacterial strain has the production capacity of long-chain QS.
Embodiment 3, pressure Shewanella (Shewanella piezotolerans) application in algal control for the 34#
First, the method for pressure Shewanella (Shewanella piezotolerans) 34# algal control
1st, the High Density Cultivation of pressure Shewanella (Shewanella piezotolerans) 34#
The pressure Shewanella 34# being obtained by embodiment 1 is carried out one-level culture (2216E liquid in 10mL triangular flask
Culture medium, formula：Peptone 5.0g, yeast extract 1.0g, ferric phosphate 0.01g, Chen Haishui 1000mL, adjust ph to 7.8), cultivate bar
Part is 30 DEG C, and rotating speed 200r/min, time 6h treat bacterial strain density close to OD600When=0.5, proceed in 100mL triangular flask and carry out
Two grades of cultures.Continue culture 18 hours, reach 1 × 10 to bacterial strain density9Individual/ML concentration, standby.
2nd, the culture of experiment frustule
Algae kind used by experiment is Alexandrium (Alexandrium catenella) (Hong Kong strain).Take laboratory passage
(initial density is 0.2 × 10 to the algae kind of culture4Individual/mL), it is sub-packed in the triangular flask of 15 300mL (often bottled liquid 100mL),
Continuous monitoring is cultivated, when algae is in exponential phase early stage, that is, algae density is 0.4 × 10 after packing4Individual/mL (train in algae by algae
Density in nutrient solution) when, carry out following group experiment, test sets five experimental group, and every group 3 are parallel.
The condition of culture of algae is as follows：Temperature is 21 ± 1 DEG C, light application time L: D=12h: 12h, intensity of illumination 3000Lx.
Five experimental group are as follows respectively：
Blank group (without 34# bacterium solution, without the culture medium for strain culturing), continues culture；
Control group (containing strain cultures, without 34# bacterium solution), final concentration of 0.05% (v/v) of nutrient solution, continues training
Support；
Strain density 1.0 × 104Individual/ML group：To in triangular flask plus by above-mentioned 1 inoculum obtaining, make bacterial density
Final concentration of 1.0 × 104Individual/ML, continues culture；
Strain density 1.0 × 105Individual/ML group：To in triangular flask plus by above-mentioned 1 inoculum obtaining, make bacterial density
Final concentration of 1.0 × 105Individual/ML, continues culture；
Strain density 1.0 × 106Individual/ML group：To in triangular flask plus by above-mentioned 1 inoculum obtaining, make bacterial density
Final concentration of 1.0 × 106Individual/ML, continues culture.
Above-mentioned each group is added various materials, is denoted as continuing the 0th day of culture.
2nd, detect
1st, the inhibition to algae for the interpolation of bacterium
By above-mentioned 5 groups of cultured products, under binocular microscope, (4 × 10 times) carry out frustule counting, each sample count 3
Secondary, results averaged.
It is found that in blank group and control group, frustule is uniformly distributed, algae culturing liquid is more limpid, in light yellow；
And when there being high concentration 34# bacterial strain to add, to when cultivating 46 days, frustule nutrient solution starts to assume muddy shape, part frustule
Dissolve, sink to the bottom.
Fig. 1 be various dose bacterium to the effect curve of algae Effects of Density it can be seen that
Blank group 0,2,4,6,8,10,12,14 days algae density (individual/mL) be respectively 4000,4066,4200,4534,
5734th, 8000,10000 and 10256.
Control group 0,2,4,6,8,10,12,14 days algae density (individual/mL) be respectively 4066,4100,4134,4466,
5534th, 7333,9000 and 8520.
Strain density 1.0 × 104Individual/ML group 0,2,4,6,8,10,12,14 days algae density (individual/mL) be respectively 4033,
4106th, 4200,4533,4800,5933,6400 and 6314.
Strain density 1.0 × 105Individual/ML group 0,2,4,6,8,10,12,14 days algae density (individual/mL) be respectively 4133,
4134th, 3600,2800,2000,1666,1002 and 762.
Strain density 1.0 × 106Individual/ML group 0,2,4,6,8,10,12,14 days algae density (individual/mL) be respectively 4166,
3466、2000、800、668、668、586、370.
As can be seen that from the beginning of 0 day microscope inspection, Shewanella (Shewanella sp.) killing to frustule for the 34#
Effect assumes dose relationship, and the raising killing effect with concentration is more notable, and killing rate reaches more than 80% within the 6th day, effect of algae restraint
Reach peak value.
2nd, the impact to Alexandrium chlorophyll and photosynthetic efficiency for the Shewanella (Shewanella sp.)
Detect that above-mentioned 5 groups are continued the chlorophyll of algae of culture, photosynthetic efficiency, concrete grammar is as follows：The leaf of each group red tide algae
Green element and photosynthetic efficiency measurement are carried out using phytoplankton classification luminoscope (PHYTO-PAM), and concrete operations are as follows, take 3mL algae
Liquid (cultured products of continuous culture) loads measuring cup and is placed in magazine, carries out 20min dark adaptation to frond, opens Phyto-
PAM modulation pulse fluoro instrument wavelength is 0.1 μm of ol/ (m for 520nm intensity2S) green test light.Measurement process by
Phytowin software controls, and opens measurement light (ML), opens saturation pulse key, write down Fv/Fm value, as after optical signal is stable
Photosynthetic efficiency yield value.(ChI) of chlorophyll levels is measured and is also carried out using this instrument.
As shown in Figures 2 and 3, wherein, Fig. 2 is the impact to Alexandrium chlorophyll content for the 34# bacterial strain to result, Fig. 3
For the impact to Alexandrium photosynthetic efficiency for the 34# bacterial strain；
From figure 2 it can be seen that
Blank group 0,2,4,6,8,10,12,14 days chlorophyll content (μ g/L) be respectively 15.21,18.78,
28.14、34.64、34.07、38.92、36.50、37.20.
Control group 0,2,4,6,8,10,12,14 days chlorophyll content (μ g/L) be respectively 16.57,19.42,
27.64、34.14、35.50、39.35、38.90、37.60.
Strain density 1.0 × 104Individual/ML group is respectively in the chlorophyll content (μ g/L) of 0,2,4,6,8,10,12,14 days
15.85、19.07、27.50、34.57、34.35、38.00、36.50、37.19.
Strain density 1.0 × 105Individual/ML group is respectively in the chlorophyll content (μ g/L) of 0,2,4,6,8,10,12,14 days
15.64、16.71、12.00、10.92、11.35、11.28、9.50、7.21.
Strain density 1.0 × 106Individual/ML group 0,2,4,6,8,10,12,14 days chlorophyll content (μ g/L) be respectively
16.42、15.78、8.07、7.57、7.42、7.0、5.60、4.11.
As can be seen that dense (the strain density 1.0 × 10 of high dose bacterium6Individual/ML group) compared to control group, from the beginning of the 4th day, its
Chlorophyll levels are significantly lower than control group, and its value only has the 15.533.6% (P ＜ 0.05) of control group.
From figure 3, it can be seen that blank group 0,2,4,6,8,10,12,14 days photosynthetic efficiency (Fv/Fm) be respectively
0.45、0.47、0.46、0.57、0.60、0.61、0.63、0.62.
Control group 0,2,4,6,8,10,12,14 days photosynthetic efficiency (Fv/Fm) be respectively 0.46,0.48,0.47,
0.56、0.63、0.64、0.64、0.65.
Strain density 1.0 × 104Individual/ML group is respectively in the photosynthetic efficiency (Fv/Fm) of 0,2,4,6,8,10,12,14 days
0.45、0.46、0.50、0.58、0.63、0.64、0.58、0.60.
Strain density 1.0 × 105Individual/ML group is respectively in the photosynthetic efficiency (Fv/Fm) of 0,2,4,6,8,10,12,14 days
0.47、0.47、0.48、0.35、0.35、0.38、0.34、0.34.
Strain density 1.0 × 106Individual/ML group 0,2,4,6,8,10,12,14 days photosynthetic efficiency (Fv/Fm) be respectively 0.46,
0.40、0.40、0.30、0.31、0.31、0.29、0.30.
As can be seen that external source adds the bacterium (10 of high dose6Individual/ML), frustule photosynthetic efficiency level is relatively low, and compares
Group then has an obvious uphill process.
This shows, the presence of bacterium, to the physiology of algae bring stress pressure, the energy of algae itself is used for tackling ring
Border is coerced, thus reducing its ability to luminous energy capture.
Claims (10)
1. pressure Shewanella (Shewanella piezotolerans) 34#, its preserving number is CGMCC NO.6490.
2. application in preparing colony induction signaling for the pressure Shewanella 34# described in claim 1.
3. application according to claim 2, the method wherein preparing colony induction signaling comprises the steps：
1) ferment pressure Shewanella 34#, and supernatant is collected by centrifugation；
2) it is extracted with ethyl acetate described supernatant, collect organic phase, that is, obtain colony induction signaling extract.
4. according to claim 3 application it is characterised in that：
Step 1) in, the temperature of described fermentation is 30 DEG C, and the time of described fermentation is 48h；The rotating speed of described centrifugation is 8000r/
Min, the time of described centrifugation is 20min, and described centrifugation radius is 13.5cm；
Step 2) in, described supernatant and ethyl acetate extraction volume ratio are 1: 2, and time of described extraction is 3h.
5. the application according to claim 3 or 4 it is characterised in that：
In step 2) in, also include successively removing the ethyl acetate and dissolving in described organic phase after described collection organic phase
Step；
In described step 1) and step 2) between also include the step that filters described supernatant.
6. the colony induction signaling extract prepared by the method described in claim 3-5 any one.
7. application in preventing and treating red tide for the pressure Shewanella 34# described in claim 1.
8. according to claim 7 application it is characterised in that：Described preventing and treating red tide passes through to suppress red tide algae propagation to realize.
9. the application according to claim 7 or 8 it is characterised in that：Described suppression red tide algae growth is embodied in following 1) -4)
In 4 kinds or any of which：
1) reduce red tide algae density；
2) reduce the chlorophyll content of red tide algae；
3) reduce the photosynthetic efficiency of red tide algae；
4) change the composition of red tide algae algae border microorganism.
10. according to claim 9 application it is characterised in that：Described application is to add in the growth system of red tide algae
Pressure Shewanella 34# described in claim 1, to suppress red tide algae to grow, described red tide algae comprises or part is gone through for Asia
The big algae in mountain.
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| CN201210596277.6A CN103173383B (en) | 2012-12-26 | 2012-12-26 | Shewanella piezotolerans 34# and application thereof to algae inhibition |
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| CN103509744B (en) * | 2013-09-11 | 2015-07-15 | 上海交通大学 | Algicidal Shewanella and application thereof in controlling blue algae water bloom |
| CN106085896B (en) * | 2016-05-10 | 2019-07-09 | 清华大学深圳研究生院 | Enterobacter cloacae and application thereof |
| CN111944710B (en) * | 2019-05-15 | 2022-10-04 | 东北林业大学 | Shewanella and application thereof in improving stress resistance of plants |
| CN114908013B (en) * | 2022-05-19 | 2023-01-31 | 中国水产科学研究院黑龙江水产研究所 | Shewanella manshurica for producing DDP-IV inhibitor and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1621513A (en) * | 2003-11-28 | 2005-06-01 | 广东省微生物研究所 | Shewanella decolorationis |
| CN101880638A (en) * | 2009-05-06 | 2010-11-10 | 国家***第三海洋研究所 | Shewanella and application thereof in microbiological fuel cell |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1621513A (en) * | 2003-11-28 | 2005-06-01 | 广东省微生物研究所 | Shewanella decolorationis |
| CN101880638A (en) * | 2009-05-06 | 2010-11-10 | 国家***第三海洋研究所 | Shewanella and application thereof in microbiological fuel cell |
Non-Patent Citations (1)
| Title |
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| 藻细菌鉴定及溶藻机理的初步研究;曹宾霞等;《全国海洋生物技术与海洋药物学术会议论文集》;20060806;616-619 * |
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