CN105400703A - Paecilomyces sp. fungus and application thereof - Google Patents
Paecilomyces sp. fungus and application thereof Download PDFInfo
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Abstract
The invention discloses a Paecilomyces sp. fungus and application thereof. The Pasecilomyces sp. fungus is named paecilomyces QD10 and preserved in the China Center for Type Culture Collection, the address is in Wuhan university, the microorganism preservation number is CCTCC NO.M2015405, and the preservation date is June 26th, 2015. The minimum inhibitory concentration (MIC) of the Pasecilomyces sp. fungus QD10 is cadmium (Cd<2+>)1000 mg/L, and the common resistance is 800&1500 mg/L (Cd<2+>&Pb<2+>). The adsorption effect can be improved by researching the adsorption condition, the adsorption capacity is improved, the self stress resistance of a high-resistance QD10 strain is stronger, and therefore the Paecilomyces sp. fungus can be applied to heavy metal cadmium or lead pollution treatment.
Description
Technical field
The present invention relates to a strain paecilomyces fungi, particularly strain paecilomyces fungi QD10 bacterial strain and uses thereof.The invention belongs to microbial technology field.
Background technology
Along with the growth at full speed of world economy, economical activities of mankind increases, and brings the random process of large problem-environmental pollution, particularly industrial waste discharges that a whole world can not be ignored, agrochemicals, very large pollution is caused, especially heavy metal contamination to soil, river etc.So far, lead about 5 × 10 is discharged in the whole world every year on average
5ten thousand kg, mercury about 1,500 ten thousand kg etc.Also there are some researches show, some primary agricultural products come into the market have heavy-metal residual thing, as cadmium rice etc.These enter food chain, can give physical environment even human body cause very large damage.Therefore, carrying out of heavy metal contamination repair has very great meaning for the whole mankind and ecotope.
The control of heavy metal contamination is paid close attention in many ways, especially to the soil remediation of plumbous cadmium pollution.Plumbous, cadmium belongs to five large polluted heavy metals, and its pollution range is very wide, and water body, soil etc. are all by its evil.Particularly soil, pollution range was more and more wider in the last few years.According to statistics, China about has 1.3 × 10
4hm
2arable land is subject to lead and waits heavy metal severe contamination, causes grain drop in production to reach 1.0 × 10
7t.The use of doped fuel has also increased the weight of the pollution of Lead In Soil.In recent years, in the testing process to edible farm crop such as vegetables, found that there is lead, phenomenon that cadmium exceeds standard.
The harm that the pollution of Pb in Soil cadmium brings is very large, all has a great impact the mankind and ecotope.In plant, first can cause production declining, serious plant can be dead.In humans and animals, plumbous, the murder by poisoning of cadmium to animal and human is entered in human body and animal body by food chain.The excessive meeting of Pb accumulation causes degradation toxicity symptom under kidney failure, intelligence.Cd accumulation is excessive may be caused " itai-itai ", and cause the diseases such as glomerular function exhaustion, affect the activity in production of animal, serious heavy metal can spread all over animal whole body.
Compared with microorganism remediation technology, common physical chemistry recovery technique easily causes secondary pollution.And the heavy metal Land pollution recovery technique that microorganism remediation becomes popular to pollute the feature such as little, cost is low, more and more receive publicity.Microorganism remediation heavy-metal contaminated soil and water body refer to and alleviate the harm of heavy metal for environment by the interaction between organism and heavy metal particles.In the biomaterial that biological renovation method is used, fungal material has the many merits such as biomass is large, heavy metal adsorption capacity large, the rear metal of absorption easily reclaims.And microbial metabolism material is as exocellular polysaccharide, chitin etc. also participate in absorption and the immobilization process of heavy metal.This is mainly based on the functional group of microbial cell surface, and as hydroxyl, carbonyl, sulfydryl and amine groups etc., these functional groups can be combined effectively with heavy metal cation, thus have the adsorption of heavy metal.The biological adsorption of fungi heavy metal is the emphasis of Pollution abatement research and utilization.
The present invention obtains strain paecilomyces (Paecilomycessp.) fungi QD10 bacterial strain through screening and identification, and the minimum inhibitory concentration (MIC) of this bacterial strain is cadmium (Cd
2+) 1000mg/L, common resistance is 800 & 1500mg/L (Cd
2+aMP.AMp.Amp Pb
2+).Improve its adsorption effect by research adsorption conditions, improve adsorptive power, make resistance himself resistance of QD10 bacterial strain stronger, be more conducive to practical application.
Summary of the invention
An object of the present invention is to provide a strain paecilomyces fungi, and the minimum inhibitory concentration (MIC) of this fungi is cadmium (Cd
2+) 1000mg/L, common resistance is 800 & 1500mg/L (Cd
2+aMP.AMp.Amp Pb
2+).
Two of object of the present invention is to provide the described application of paecilomyces fungi in heavy metal cadmium or control of Pb pollution.
In order to achieve the above object, present invention employs following technique means:
Strain paecilomyces (Paecilomycessp.) fungi that the present invention screens, called after Paecilomyces varioti QD10 (Paecilomycessp.QD10), Classification And Nomenclature is Paecilomyces varioti QD10 (Paecilomycessp.QD10), China typical culture collection center is deposited on June 26th, 2015, address is in Wuhan University, and its microbial preservation number is CCTCCNO.M2015405.
Paecilomyces fungi QD10 bacterial strain has following characteristic:
Microorganism feature: bacterium colony surface is in white on solid medium, and reverse side is faint yellow, quality flocculence, fast, when solid culture basal growth 48h, its diameter can 2-3cm in growth.Crystallization is there will be in cultivation 10d rear surface.Show QD10 vegetative hyphae wall under opticmicroscope smooth, hyphae colorless is transparent, and mycelia is flourishing, and top is tapered into obvious elongated necks, visible a large amount of elliptic spore.
Physico-chemical property: well-grown in pH3-9 potato (PDA) substratum, well-grown between temperature 10-45 DEG C, can make gelatine liquefication, hydrolyzed starch and Mierocrystalline cellulose, and maltose and Mierocrystalline cellulose can also be utilized as carbon source.
The ITSs-rDNA sequence of paecilomyces fungi QD10 bacterial strain of the present invention and the ITSs-rDNA sequence of the blue or green Pseudomonas fungi of the plan delivered at present, homology reaches more than 98%.Comprehensive morphological feature and molecular biology identification, QD10 bacterial strain is paecilomyces (Paecilomycessp.) fungi, and its ITSs-rDNA conserved sequence is as shown in SEQIDNO:1.
The minimum inhibitory concentration (MIC) of paecilomyces fungi QD10 bacterial strain of the present invention is cadmium (Cd
2+) 1000mg/L, common resistance is 800 & 1500mg/L (Cd
2+aMP.AMp.Amp Pb
2+).Improve its adsorption effect by research adsorption conditions, improve adsorptive power, make resistance self resistance of QD10 bacterial strain comparatively strong, be more conducive to practical application.
Therefore, further, present invention also offers the described application of paecilomyces fungi in heavy metal cadmium or control of Pb pollution.Preferably, by described paecilomyces fungi and plant coupling, jointly for the Pollution abatement of heavy metal cadmium or lead.
In sum, the paecilomyces fungi QD10 bacterial strain mixing better resistance that the present invention filters out, at 800 & 1500mg/L (Cd
2+aMP.AMp.Amp Pb
2+) mixed culture medium in also can well grow.Improve its adsorption effect by research adsorption conditions, improve adsorptive power, make resistance self resistance of QD10 bacterial strain stronger, be more conducive to practical application.
Accompanying drawing explanation
Fig. 1 is the colony characteristics of QD10 bacterial strain;
Fig. 2 is the QD10 vegetative hyphae and spore shape that show under opticmicroscope;
Fig. 3 is that difference throws bacterium amount to the effect diagram of QD10 bacterial strain adsorption effect, and wherein A figure is that different bacterium amount of throwing is to QD10 bacterial strain absorption Cd
2+the effect diagram of effect, B figure is that different bacterium amount of throwing is to QD10 bacterial strain absorption Pb
2+the effect diagram of effect;
Fig. 4 is the effect diagram of different pH value to QD10 bacterial strain adsorption effect;
Fig. 5 is the effect diagram of differing temps to QD10 bacterial strain adsorption effect;
Fig. 6 is the effect diagram of different time to QD10 bacterial strain adsorption effect;
Fig. 7 adopts QD10 bacterial strain or the effect with plant combined remediating heavy metal cadmium or lead pollution of soil.
Embodiment
Further describe the present invention below in conjunction with specific embodiments and the drawings, advantage and disadvantage of the present invention will be more clear along with description.But embodiment is only exemplary, does not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments and replacement all fall within the scope of protection of the present invention.
The separation of embodiment 1 paecilomyces fungi of the present invention QD10 bacterial strain, Isolation and characterization
1 materials and methods
1.1 substratum and solution
Potato (PDA) substratum: glucose 20g, the potato leach liquor 1000ml of about 20% (takes peeled potatoes 200g, stripping and slicing, the 1000ml that adds water boils half an hour, crosses leaching juice with double gauze, supplies the water yield to 1000ml), agar 20g, pH nature.
Basic medium: glucose 10g, saltpetre 1g, magnesium sulfate 0.2g, potassium primary phosphate 0.5g, agar 15g, distilled water 1000ml.
Gelatine liquefication substratum: peptone 5g, gelatin 200g, glucose 20g, distilled water 1000ml.
Starch Hydrolysis substratum: K
2hPO
410.0g, KH
2pO40.3, NaCl0.5g, MgCO
31.0g, KNO
31.0g, Zulkovsky starch 2.0g, agar 15g; PH7.2, distilled water 1000ml.
Cellulose hydrolysis substratum: MgSO
47H
2o0.5g, K
2hPO
40.5g, NaCl0.5g, KNO
31.0g, filter paper bar, distilled water 1000ml.
Solution: preparation 100gL
-1pb (NO
3)
2mother liquor and 100gL
-1cdSO
4mother liquor, sterilizing is for subsequent use.
1.2QD10 the separation and purification of bacterial strain:
1. soil sampling 10g loads in the triangular flask of 90mL sterilized water, makes soil supension, respectively get 0.1mL10 at 28 DEG C and 150r/min shaking table vibration 2h postprecipitation 30min
-1, 10
-2, 10
-3, 10
-4, 10
-5the soil sample suspension of concentration, is coated on screening solid medium (PDA substratum) flat board with coating method, cultivates 4-5d for 28 DEG C, then picking individual colonies, is separated, through repeatedly purifying with same method line, obtain single strain, the bacterial classification of purifying is put into 4 DEG C of refrigerators and save backup.
2. by inoculation good for purifying in the PDA substratum having heavy metal, Cd in substratum
2+ion and Pb
2+the concentration of ion respectively with 100,200,400,600,800,1000mg/L and 200,400,600,800,1000, the gradient of 1500mg/L rules successively, selects the good bacterial strain of heavy metal tolerance.
3. the bacterial strain selected is linked in the metal ion substratum of mixing, Cd in substratum
2+ion and Pb
2+the concentration of ion is that the gradient of 100 & 400,200 & 600,400 & 800,800 & 1000,800 & 1500mg/L is rule successively, selects the bacterial strain of better resistance.
Through above-mentioned screening step, filter out the QD10 bacterial strain that a strain resistance is the strongest.Wherein the minimum inhibitory concentration (MIC) of this bacterial strain is cadmium (Cd
2+) 1000mg/L, common resistance is 800 & 1500mg/L (Cd
2+aMP.AMp.Amp Pb
2+).
The microbial morphology of 1.3QD10 bacterial strain and physico-chemical property are identified
1.3.1QD10 the microbial morphology of bacterial strain
Observe colony characteristics, examine under a microscope spore and mycelia feature.
1.3.2QD10 the physical and chemical property determining of bacterial strain
Gelatine liquefication measures: inoculation test strain QD10 bacterial strain is in gelatin surface, and light culture at 28 DEG C, observes once every 5d.First by test tube subcooling, observed and recorded after gelatin in nonvaccinated CK test tube solidifies.Do not liquefy as then cooled rear gelatin without protease hydrolysis effect, otherwise then show there is protease hydrolysis effect.
Starch Hydrolysis measures: poured into by substratum in culture dish, adopts the inoculation of some connection, at 28 DEG C after light culture 10 ~ 20d, pour the iodine liquid prepared into media surface after culture medium solidifying.Produce if any amylase, then the surrounding of bacterium colony can not become blueness, otherwise then becomes blue, shows have amylase to produce.The transparent circle formed is utilized to detect whether produce amylase.
Maltose or cellulose utilization measure: change the carbon source glucose in basic medium into maltose or Mierocrystalline cellulose, all the other medium components are constant, and can observe QD10 bacterial strain normal growth.
Cellulose hydrolysis measures: be placed in the substratum of test tube by a filter paper bar, in half immersion substratum, half is on substratum, test strains is inoculated on the filter paper bar in sterilized test tube, after 28 DEG C of cultivation 30d, observes it and whether can grow on filter paper bar.
Growth temperature and pH scope measure: by QD10 inoculation PDA substratum, at different temperatures, cultivate 7 days, collected by centrifugation bacterial strain, the bacterial strain collected is placed in 65 DEG C of bakings and weighs after 4 hours, according to the bacterial strain dry weight (g/L) of this bacterium, and the growth temperature range of this bacterium known;
Under optimum growth temp 30 DEG C of conditions, in the same way, measure bacterial strain dry weight, according to the bacterial strain dry weight (g/L) of this bacterium, the pH scope of the growth of this bacterium known.
1.3.3QD10 the molecular biology identification of bacterial strain
The first step: extract QD10 bacterial strain DNA
(1) get fungus culture medium 1-5ml, the centrifugal 1min of 10,000rpm, exhaust supernatant as far as possible;
(2) in bacterial strain precipitation, add 200 μ l damping fluid GA, vibrate to bacterial strain and thoroughly suspend;
(3) Xiang Guanzhong adds 20 μ lProteinaseK solution, mixing;
(4) add 220 μ l damping fluid GB, vibration 15sec, place 10min for 70 DEG C, solution strain is limpid, of short duration centrifugal with the globule removing cap wall;
(5) add 220 μ l dehydrated alcohols, fully, now may there is flocks in vibration mixing 15sec, of short duration centrifugal with the globule removing cap wall;
(6) all add in an adsorption column CB3 (adsorption column puts into collection tube) by previous step gained solution and flocks, the centrifugal 30sec of 12,000rpm, outwells waste liquid, and adsorption column CB3 is put into collection tube;
(7) in adsorption column CB3, add 500 μ l damping fluid GD, the centrifugal 30sec of 12,000rpm, outwells waste liquid, and adsorption column CB3 is put into collection tube;
(8) in adsorption column CB3, add 600 μ l rinsing liquid PW, the centrifugal 30sec of 12,000rpm, outwells waste liquid, and adsorption column CB3 puts into collection tube;
(9) repetitive operation step (8);
(10) put back in collection tube by adsorption column CB3, the centrifugal 2min of 12,000rpm, outwells waste liquid, adsorption column CB3 is placed in room temperature and places several minutes, thoroughly to dry rinsing liquid remaining in sorbing material;
(11) proceeded to by adsorption column CB3 in a clean centrifuge tube, the unsettled dropping in the middle part to adsorption film 50-200 μ l elution buffer TE, room temperature places the centrifugal 2min of 2-5min, 12,000rpm, by solution collection in centrifuge tube.
Second step: use universal primer to carry out pcr amplification 18s-rDNA fragment, the PCR primer after amplification is carried out race gel electrophoresis, checks band.
Reaction system: 10ul system.Wherein 5ulMasterMix, forward primer and reverse primer each 0.2ul, ddH
2o-1.6ul, bacterium DNA masterplate 3ul.
3rd step: cut glue and reclaim purified pcr product
1. add the sol solutions of 500ul, 55 DEG C of water-bath 10min, (timing register timing) makes blob of viscose melt completely, and every 2min puts upside down mixing once;
2. during object fragment ﹤ 500bp, the Virahol of 100ul is added, mixing.55 DEG C of water-bath 1min, mixing.During object fragment >500bp, this step can be omitted;
3. shift: the Agarose liquid dissolved is moved into adsorption column, the centrifugal 40sec of 12000rpm.Outwell the liquid in collection tube, then adsorption column is put into same collection tube;
4. in adsorption column, 500ul rinsing liquid is added, the centrifugal 40sec of 12000rpm.Outwell the liquid in collection tube, then adsorption column is put into same collection tube.If room temperature is lower than 20 DEG C, before centrifugal, first leave standstill 1min;
5. in adsorption column, 500ul rinsing liquid is added, the centrifugal 40sec of 12000rpm.Outwell the liquid in collection tube, adsorption column is put into same collection tube;
6. the centrifugal 2min of 12000rpm, leaves standstill 10min;
7. adsorption column is put into a clean 1.5ml centrifuge tube.Add 30ulTE elutriant (according to electrophoretic band brightness decision) in adsorption film central authorities, leave standstill the centrifugal 2min of 5min, 12000rpm.
4th step: by BigDyeterminator test kit processing sample, use ABI3730DNA sequenator to carry out capillary electrophoresis, is detected the fluorescence chemical mark of DNA in sample, is obtained the sequence of sample DNA by data processing.
1.4QD10 bacterial strain adsorption effect at different conditions
QD10 bacterial strain is measured at different conditions its heavy metal adsorption effect, comprising temperature, pH value, connect bacterium amount and time, investigate the adsorption effect of bacterial strain.
1.4.1 different bacterium amount of throwing is on the impact of QD10 bacterial strain adsorption effect
In the triangular flask of 250ml, add the deionized water of 100ml, in triangular flask, add the Cd that concentration is 50mg/L
2+with the Pb of 100mg/L
2+solution, temperature is 30 DEG C, and pH value is nature, add different amount mycelia (0g, 0.2g, 0.4g, 0.6g, 0.8g, 1.0g, 1.2g, 1.4g) shake 60min after centrifugal, get supernatant liquor, measure Cd in supernatant liquor
2+and Pb
2+concentration, the different bacterium amount of throwing of research is on the impact of QD10 bacterial strain adsorption effect.
1.4.2 different pH value is on the impact of QD10 bacterial strain adsorption effect
Fungal growth situation and environment acid-basicity closely related, and usually lower in cadmium and Lead pollution environment pH value, therefore study this bacterial strain and whether possess the ability adapting to sour environment.In the triangular flask of 250ml, add the deionized water of the 100ml of different pH value (pH value is 3,4,5,6,7,8,9), in triangular flask, add the Cd that concentration is 50mg/L
2+with the Pb of 100mg/L
2+solution, temperature is 30 DEG C, centrifugal after adding 0.2g mycelia concussion 60min, gets supernatant liquor, measures Cd in supernatant liquor
2+and Pb
2+concentration, study the impact of different pH value on QD10 bacterial strain adsorption effect.
1.4.3 differing temps is on the impact of QD10 bacterial strain adsorption effect
In microbial growth process, temperature affects the dissolving etc. of the activity of enzyme, the mobility of cytoplasmic membrane and material, thus causes its growth velocity and adsorption effect to change.In the triangular flask of 250ml, add the deionized water of 100ml, in triangular flask, add the Cd that concentration is 50mg/L
2+with the Pb of 100mg/L
2+solution, differing temps (15 DEG C, 20 DEG C, 25 DEG C, 30 DEG C, 30 DEG C, 40 DEG C), pH value is nature, centrifugal after adding 0.2g mycelia concussion 60min, gets supernatant liquor, measures Cd in supernatant liquor
2+and Pb
2+concentration, study the impact of different pH value on QD10 bacterial strain adsorption effect.
1.4.4 different time is on the impact of QD10 bacterial strain adsorption effect
In the triangular flask of 250ml, add the deionized water of 100ml, in triangular flask, add the Cd that concentration is 50mg/L
2+with the Pb of 100mg/L
2+solution, temperature is 30 DEG C, and pH value is nature, adds 0.2g mycelia concussion different time (0min, 20min, 40min, 60min, 80min, 100min, 120min, 140min, 160min, 180min, 200min) centrifugal afterwards, get supernatant liquor, measure Cd in supernatant liquor
2+and Pb
2+concentration, study the impact of different pH value on QD10 bacterial strain adsorption effect.
2 results
The microbial morphology of 2.1QD10 bacterial strain and physico-chemical property qualification result
2.1.1QD10 the microbial morphology of bacterial strain
The colony characteristics of QD10 bacterial strain is shown in Fig. 1, and on solid medium, bacterium colony surface is in white (Figure 1A), and reverse side is faint yellow (Figure 1B), quality flocculence, and fast, when solid culture basal growth 48h, its diameter can 2-3cm in growth.Crystallization is there will be in cultivation 10d rear surface.Show QD10 vegetative hyphae wall under opticmicroscope smooth, hyphae colorless is transparent, and mycelia is flourishing, and top is tapered into obvious elongated necks, and visible a large amount of elliptic spore, shown in Fig. 2.
2.1.2QD10 the physico-chemical property qualification result of bacterial strain
The physico-chemical property qualification result of QD10 bacterial strain is in table 1.
The physico-chemical property qualification result of table 1QD10 bacterial strain
| Title | Result |
| Gelatine liquefication | + |
| Starch Hydrolysis | + |
| Cellulose hydrolysis | + |
| Maltose utilizes | + |
| Cellulose utilization | + |
| pH | 3-9 |
| Temperature | 10-45℃ |
2.1.3QD10 the molecular biology identification result of bacterial strain
Molecular biology identification found that: the ITSs-rDNA sequence (shown in SEQIDNO:1) of QD10 bacterial strain and the ITSs-rDNA sequence of the paecilomyces fungi of delivering at present, homology reaches more than 98%.Comprehensive morphological feature and molecular biology identification, QD10 bacterial strain is paecilomyces (Paecilomycessp.) fungi.
The above-mentioned Strain Designation screened is Paecilomyces varioti QD10 (Paecilomycessp.QD10), and be deposited in China typical culture collection center on June 26th, 2015, address is in Wuhan University, and its microbial preservation number is CCTCCNO.M2015405.
2.2QD10 bacterial strain adsorption effect at different conditions
2.2.1 different bacterium amount of throwing is on the impact of QD10 bacterial strain adsorption effect
The results are shown in Figure 3, according to Fig. 3, the Cd when throwing bacterium amount is 0.2g to 1.0g
2+and Pb
2+the adsorption rate of two kinds of ions is all the trend that rises gradually and adsorptive capacity in downward trend gradually.When to throw bacterium amount be 0.2g to 0.6g, the adsorptive capacity change of bacterial strain obviously, be 0.6g to 1.2g is change gradually to delay in throwing bacterium amount, and tend to be steady, illustrate that mycelium quality reaches capacity when certain concentration, the adsorptive capacity of unit volume also can tend to be steady thereupon.
2.2.2 different pH value is on the impact of QD10 bacterial strain adsorption effect
The results are shown in Figure 4, according to Fig. 4, when pH value 3 to 9, adsorption rate is in the overall trend risen.When pH value is 3, adsorption rate is very low, may be due to the H3O in solution
+with Pb
2+the reason of the avtive spot of competition cell wall surface.When pH value is greater than 7 meta-alkalescence, Cd
2+and Pb
2+can with OH
-ion forms the precipitation of oxyhydroxide, and the rising of adsorption rate can not ascribe the absorption of bacterial strain completely to, so we think when pH value is 6, and Cd
2+and Pb
2+adsorption effect is best, and adsorption rate reaches 47.50% and 75.58%.
2.2.3 differing temps is on the impact of QD10 bacterial strain adsorption effect
The results are shown in Figure 5, according to Fig. 5, bacterial strain is to Cd
2+and Pb
2+absorption be all the process declined afterwards that first rises, when 25 DEG C and 30 DEG C, clearance is better.Also be the temperature that mycelial growth breeding is relatively suitable in the temperature range of 30 DEG C, so Cd
2+and Pb
2+clearance reaches maximum value 39.58% and 61.73%.
2.2.4 different time is on the impact of QD10 bacterial strain adsorption effect
The results are shown in Figure 6, according to Fig. 6, bacterial strain rate of adsorption when 0 ~ 60min is very fast, has just started a large amount of avtive spot of bacterial strain and Cd
2+and Pb
2+in conjunction with, the heavy metal in solution is adsorbed rapidly.After 60min to 100min, trend slows down gradually and trends towards balance, illustrates that a large amount of ion is combined with site, and absorption reaches capacity.
The application of embodiment 2 paecilomyces fungi QD10 bacterial strain in heavy metal cadmium or control of Pb pollution
Adopt potted plant reed to test, potted plant reed is with the seed plantation reed plant of 3-5 month.Be designed to A, B two groups, often organize 5 kinds of process, often kind of process three repetitions, 100g soil/basin.First by soil aging, the content of Cd2+ and Pb2+ is respectively: A group: 13.67mg/kg and 54.26mg/kg, B group: 22.39mg/kg and 137.8mg/kg.Use after soil disinfection, be respectively: C1 is for adding 10mlQD10 bacterium liquid, and C2 is for adding 20mlQD10 bacterium liquid, and Y is potted plant reed, K1 is that potted plant reed adds 10mlQD10 bacterium liquid, and K2 is that potted plant reed adds 20mlQD10 bacterium liquid.The whole reparation phase adds sterilized water, and repair time is 40 days.
Wherein, QD10 bacterium liquid is prepared in accordance with the following methods: 1, allow QD10 bacterial classification grow on test tube slant substratum, to growing spore, 2, after covering with spore, super clean bench operates, injects in slant tube with sterile distilled water (physiological saline, PBS damping fluid), scrape chamfered surface gently with aseptic inoculation ring, make the abundant wash-out of spore, mixing; 3, by the filter paper of liquid in step 2 by aseptic, common funnel filters (removing non-spore thing), with the small beaker of aseptic, splendid attire filtrate, gets filtrate and can obtain spore suspension, be i.e. QD10 bacterium liquid; 4, with blood counting chamber counting, concentration is 2 × 10
8-2 × 10
10cFU/ml.
Result as shown in Figure 7, comparatively obvious according to the effect of Fig. 7 known phytomicroorganism combine d bioremediation, residual gravity metal content in soil is lower, inoculating strain amount is that the effect of 20ml is better than 10ml, flora environment in not spoiled soil is described, the repairing effect connecing bacterium amount under the condition of colony balance larger is better.Microorganism is repaired separately does not have the effective of phytomicroorganism combine d bioremediation, the great role that plant plays in repair process, and microorganism can strengthen the adsorption effect of plant heavy metal in soil.
Claims (3)
1. strain paecilomyces (Paecilomycessp.) fungi, called after Paecilomyces varioti QD10, be deposited in China typical culture collection center, address is in Wuhan University, its microbial preservation number is CCTCCNO.M2015405, and preservation date is on June 26th, 2015.
2. the application of paecilomyces fungi according to claim 1 in the Pollution abatement of heavy metal cadmium or lead.
3. application according to claim 2, is characterized in that paecilomyces fungi according to claim 1 and plant coupling, jointly for the Pollution abatement of heavy metal cadmium or lead.
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| CN114874916A (en) * | 2022-03-28 | 2022-08-09 | 中国地质大学（武汉） | Scopulariopsis fungus (Sarocladium kiliense) ZJ-1 and application thereof |
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