CN103103151B - Streptomyces thermocarboxydus NL-1 as actinomycete capable of efficiently eroding limestone and application of Streptomyces thermocarboxydus NL-1 - Google Patents
Streptomyces thermocarboxydus NL-1 as actinomycete capable of efficiently eroding limestone and application of Streptomyces thermocarboxydus NL-1 Download PDFInfo
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- CN103103151B CN103103151B CN201310018611.4A CN201310018611A CN103103151B CN 103103151 B CN103103151 B CN 103103151B CN 201310018611 A CN201310018611 A CN 201310018611A CN 103103151 B CN103103151 B CN 103103151B
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Abstract
The invention discloses Streptomyces thermocarboxydus NL-1 as actinomycete capable of efficiently eroding limestone and an application of the Streptomyces thermocarboxydus NL-1. The actinomycete capable of efficiently eroding limestone is classified and named as Streptomyces thermocarboxydus NL-1 and is preserved in the China Center for Type Culture Collection (CCTCC), wherein the preservation number is CCTCC NO: M212460, the preservation date is Nov. 19, 2012 and the address of the preservation unit is Wuhan University in China. The Streptomyces thermocarboxydus NL-1 has strong erosion capability to limestone and can be used for accelerating release of main ions, such as Ca and Mg in rocks and accelerating erosion and soil formation of limestone; and the volume weight of a spray-seeding substrate can be reduced, the erosion resistance and the shearing resistance of the spray-seeding substrate are enhanced, and a favorable application and development prospect is achieved.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of limestone efficiently corrode the thermophilic carbon monoxide streptomycete of actinomycetes (
streptomyces thermocarboxydus) NL-1 and application thereof.
Background technology
Seed spraying technology be by the mixture of plant seed and growth substrate material evenly, the high pressure greening technique of sowing grass seeds by duster on ground is domatic.This technology is mainly for a kind of greening technique of the hard side slope research and development such as rock.Be widely used in the slope greening of the engineerings such as water conservancy, highway, railway at present.But this technology is directed to rock slope and realizes greening and must possess two conditions: one is scar has plant to rely the planting matrix of continued propagation; Two is that planting matrix mass-energy is fixed on scar for a long time, the erosion of blowing-resistant, heavy rain and flowing water.But in Practical Project, current Seed spraying technology, is difficult to solve above-mentioned two technical barriers, matrix of sowing grass seeds by duster is difficult to permanent maintain on rock slope, makes the utilization of this technology be subject to a lot of restrictions.Obviously matrix of sowing grass seeds by duster has become the core content of rock slope Seed spraying technology, this matrix not only requires the soil property providing suitable slope-protecting vegetation to grow, also require to there is the attachment function adhered on rock slope, the fusion completely permanent with domatic realization.
Rocky erosion microbial bacteria group energy accelerates the soil process of rock, forms the soil adapted to needed for plant life fast, simulate natural ecological environment, promotes that rocky erosion becomes the succession of soil, for plant-growth provides nutritive ingredient continuously.Under the effect of rocky erosion microorganism species, at rock mass with sow grass seeds by duster between matrix and define an interface---vegetation base.This vegetation base, both solved plant on scar to rely the planting matrix problem (comprise and nutritive element is provided) of continued propagation, also solve planting matrix mass-energy and be permanently affixed at scar, realize the fusion of scar and planting matrix, facilitate the material cycle of planting matrix and rock.Obviously, screening efficient rocky erosion microorganism strains is the key that alien earth sparging covers green technology long term maintenance and widespread use.At present, domestic for different rock, the efficient rocky erosion microorganism strains that screening is suitable for have not been reported.
Summary of the invention
Goal of the invention: for above-mentioned problems of the prior art, the object of this invention is to provide a kind of limestone and efficiently corrode actinomycetes thermophilic carbon monoxide streptomycete NL-1, the foundation of efficiently corroding microbial bacteria storehouse for rock provides bacterial classification support.Another object of the present invention is to provide the application that above-mentioned limestone efficiently corrodes actinomycetes thermophilic carbon monoxide streptomycete NL-1.
Technical scheme: in order to realize foregoing invention object, the technical solution used in the present invention is as follows:
A kind of limestone efficiently corrodes actinomycetes, its Classification And Nomenclature be thermophilic carbon monoxide streptomycete (
streptomyces thermocarboxydus) NL-1, be preserved in China typical culture collection center, deposit number CCTCC NO:M 2012460, preservation date: on November 19th, 2012; Depositary institution address: Wuhan, China Wuhan University.
Described limestone efficiently corrodes actinomycetes and is corroding the application in rock.
Described rock is lime rock.
Thermophilic carbon monoxide streptomycete NL-1 is that (it is limestone, primarily of CaCO from the exposed crag of Nanjing Technology in Mine Spoils in Mufu Mountain
3composition, is rich in Ca) surface screening obtains, and to accelerate in lime rock the Element releases such as Ca, Mg.
Thermophilic carbon monoxide streptomycete NL-1 bacterial strain main biological property: cultivate under Gause I solid medium 28 DEG C of conditions of improvement, mycelium gos deep in substratum, and fibrillae of spores becomes chain, spore is kept straight on, middle length, ovalization, surface irregularity.
Thermophilic carbon monoxide streptomycete NL-1 bacterial strain 16S rDNA gene order, is shown in shown in SEQ ID NO.1.Sequence in surveyed 16S rDNA sequence and GenBank database is carried out BLAST comparison.Result shows, thermophilic carbon monoxide streptomycete NL-1 bacterial strain with
streptomyces thermocarboxydussimilarity be 99%.Combining form feature and 16S rDNA sequential analysis, be accredited as thermophilic carbon monoxide streptomycete (
streptomyces thermocarboxydus) NL-1.
Beneficial effect: compared with prior art, the efficient erosion rock actinomycetes thermophilic carbon monoxide streptomycete NL-1 that the present invention filters out, all has very strong erosional competency to limestone, can promote the release of the leading ions such as Ca in rock, accelerates limestone and cuts out soil; Can matrix unit weight of sowing grass seeds by duster be reduced, strengthen impact-resistance and the shear resistance of matrix of sowing grass seeds by duster, there is good application and development prospect.
Accompanying drawing explanation
Fig. 1 is the sampling bacterium colony figure on the exposed crag of Technology in Mine Spoils in Mufu Mountain;
Fig. 2 is the thermophilic carbon monoxide streptomycete NL-1 bacterium colony figure on the Gause I solid medium of improvement;
Fig. 3 is thermophilic carbon monoxide streptomycete NL-1 fermentation culture 15d, adds fermented liquid and the comparative result figure not adding Soluble Ca that bacterium contrasts, Mg.
Embodiment
Below in conjunction with specific embodiment, the present invention is described further.
The acquisition of embodiment 1 bacterial strain and qualification:
Gather the bacterium colony (see figure 1) on limestone (Nanjing Technology in Mine Spoils in Mufu Mountain) naked rock surface, adopt dilution-plate method, in 28 DEG C of incubators, with Gause I substratum (the 1g KNO improved
3, 20g Zulkovsky starch, 0.5g K
2hPO
4, 0.5g MgSO
47H
2o, 0.5g NaCl, 0.01g FeSO
4, 18g agar powder, 1000mL distilled water, 3% potassium bichromate 1mL is added in every 300mL substratum) cultivate 4 ~ 7d, picking list bacterium colony, purifying of repeatedly ruling, and obtained strains being inoculated in respectively in the Gause I medium slant of improvement, 4 DEG C of Refrigerator stores, obtain pure bacterial strain.This bacterial strain main biological property: cultivate (see figure 2) under Gause I substratum 28 DEG C of conditions of improvement, mycelium gos deep in substratum, and fibrillae of spores becomes chain, spore is kept straight on, middle length, ovalization, surface irregularity.
The pure inoculation that takes a morsel is in the 250mL triangular flask of Gause I liquid nutrient medium that 50mL improvement is housed, and 28 DEG C, 72h is cultivated in 180r/min concussion.Conventionally, bacterial strain DNA sequencing is extracted.Obtain 16S rDNA gene order, see shown in SEQ ID NO.1.Sequence in surveyed 16S rDNA sequence and GenBank database is carried out BLAST comparison.Result shows, this bacterial strain with
streptomyces thermocarboxydussimilarity be 99%.Combining form feature and 16S rDNA sequential analysis, be accredited as thermophilic carbon monoxide streptomycete (
streptomyces thermocarboxydus) NL-1.
The limestone erosional competency of embodiment 2 bacterial strain:
Take a morsel pure inoculation in the 250mL triangular flask of Gause I liquid nutrient medium that 50mL improvement is housed with inoculating needle, and 28 DEG C, 72h is cultivated in 180r/min concussion, obtains activated spawn.
Get above-mentioned bacterium liquid 2mL, be inoculated in and 50mL fermention medium (1g KNO is housed
3, 20g Zulkovsky starch, 0.5g K
2hPO
4, 0.5g MgSO
47H
2o, 0.5g NaCl, 0.01g FeSO
4, 10g limestone powder (cleaned Technology in Mine Spoils in Mufu Mountain rock, cross 200 mesh sieves, main related chemical constituents is in table 1), distilled water 1000mL, pH 7.0 ~ 7.5) 250mL triangular flask in, 25 DEG C, 180r/min shakes cultivation.
The main related chemical constituents of table 1 Technology in Mine Spoils in Mufu Mountain limestone
| Composition | CaO | MgO | P 2O 5 | K 2O |
| Content (%) | 50.02 | 4.17 | 0.68 | 0.31 |
Fermentation culture 15d, pours into bacteria suspension in the centrifuge tube of sterilizing, 4 DEG C, and the centrifugal 10min of 5000r/min, uses asepsis injector Aspirate supernatant, after biofilter (filter sizes is 0.22um) filters, obtain bacteria-free filtrate.
Get after above-mentioned bacteria-free filtrate 20mL adds 3 concentrated nitric acid acidifyings, utilize the ICP Optima 4300DV(U.S.) Soluble Ca, Mg concentration in test soln.The result of thermophilic carbon monoxide streptomycete NL-1 as shown in figure 3 and table 2, compared with contrasting with the fermented liquid not adding bacterium, add Soluble Ca in bacterium secondary fermentation liquid, Mg concentration be respectively do not add bacterium contrast 21.4,4.1 times.
Table 2 is thermophilic carbon monoxide streptomycete NL-1 is to the Element release of limestone
| Process | Calcium (mg/L) | Magnesium (mg/L) |
| Add bacterium | 381.11 | 129.47 |
| Contrast | 17.81 | 31.65 |
Note: P < 0.05.
The judgement criteria of actinomycetes to rocky erosion ability is there is no due to domestic at present, the beautiful monarch of the present invention's reference fourth waits (2009) about rock microorganism to related documents and our experimental datas thereof such as the weathering researchs of In Carbonate Rock, determine that actinomycetes are more than 4 times that do not add bacterium contrast for the concentration be discharged in nutrient solution such as principal element Ca of limestone, be namely defined as the efficient erosion actinomycetes strain of limestone.Visible, this strain bacterium has very strong erosional competency to limestone, can promote the release of the leading ions such as Ca, Mg in rock, accelerates limestone and cuts out soil, is that very promising efficient lime rock corrodes actinomycetes strain.
Embodiment 3 bacterial strain is on the impact of matrix of sowing grass seeds by duster:
By after bacterial strain activation (method is with embodiment 2), matrix (timbered soil of sowing grass seeds by duster is admixed according to 100mL/kg, add humic acid organic fertilizer 25%, limestone powder (crossing 100 mesh sieves) 4%, xylon 0.3 % that sows grass seeds by duster, water-holding agent 0.3 %, afforest tackiness agent 0.03% of sowing grass seeds by duster), contrast is then at the substratum admixing same volume in matrix of sowing grass seeds by duster, carry out upper basin test (basin alms bowl diameter 17cm, dark 15cm), be placed in 25 DEG C of greenhouses, keep matrix of sowing grass seeds by duster moistening.After 4 months, core cutter method is adopted to test matrix unit weight of sowing grass seeds by duster; CC Suo Bo Paderewski anti-impact instrument is adopted to measure matrix impact-resistance of sowing grass seeds by duster; Soil shearing determinator is adopted to measure matrix shear resistance of sowing grass seeds by duster.
The quality of soil weight reflect soil, the comprehensive physical situation such as structure and fertility, directly affect water storage permeable performance and the venting capability of soil.As shown in Table 3, adding the sow grass seeds by duster unit weight of matrix of bacterium is 85.57% of contrast.Add bacterium and reduce matrix unit weight, be conducive to the permeable venting capability improving matrix.
Soil anti-scouribility refers to the physical disturbance of soil opposing runoff and the ability promoting to move down, and characterizes the power of Soil Anti-erodibility.As shown in Table 3, add the sow grass seeds by duster anti-impact index of matrix of bacterium comparatively to contrast and improve 20.79%.
The slip resistance of soil reflects the stability of the soil body.Resistance to shear is strong, illustrates that soil curing is good, can improve the stability of slope body; Otherwise soil curing is poor, the easy unstability of side slope.As shown in Table 3, add the sow grass seeds by duster slip resistance of matrix of bacterium comparatively to contrast and improve 23.4%.
Table 3 is thermophilic carbon monoxide streptomycete NL-1 is on the impact of matrix of sowing grass seeds by duster
| Process | Unit weight (g/cm 3） | Anti-impact index | Slip resistance (kPa) |
| Add bacterium | 0.83 | 2.15 | 90.7 |
| Contrast | 0.97 | 1.78 | 73.5 |
Note: P < 0.05.
SEQUENCE LISTING
<110> Nanjing Forestry University
<120> limestone efficiently corrodes actinomycetes thermophilic carbon monoxide streptomycete NL-1 and application thereof
<130> 100
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1419
<212> DNA
<213> Streptomyces thermocarboxydus
<400> 1
cgcatgctac acatgcaagt cgaacgatga agcccttcgg ggtggattag tggcgaacgg 60
gtgagtaaca cgtgggcaat ctgccctgca ctctgggaca agccctggaa acggggtcta 120
ataccggata ctgatcgcct tgggcatcct tggtgatcga aagctccggc ggtgcaggat 180
gagcccgcgg cctatcagct tgttggtgag gtaatggctc accaaggcga cgacgggtag 240
ccggcctgag agggcgaccg gccacactgg gactgagaca cggcccagac tcctacggga 300
ggcagcagtg gggaatattg cacaatgggc gaaagcctga tgcagcgacg ccgcgtgagg 360
gatgacggcc ttcgggttgt aaacctcttt cagcagggaa gaagcgaaag tgacggtacc 420
tgcagaagaa gcgccggcta actacgtgcc agcagccgcg gtaatacgta gggcgcgagc 480
gttgtccgga attattgggc gtaaagagct cgtaggcggc ttgtcgcgtc ggttgtgaaa 540
gcccggggct taaccccggg tctgcagtcg atacgggcag gctagagttc ggtaggggag 600
atcggaattc ctggtgtagc ggtgaaatgc gcagatatca ggaggaacac cggtggcgaa 660
ggcggatctc tgggccgata ctgacgctga ggagcgaaag cgtggggagc gaacaggatt 720
agataccctg gtagtccacg ccgtaaacgg tgggcactag gtgtgggcga cattccacgt 780
cgtccgtgcc gcagctaacg cattaagtgc cccgcctggg gagtacggcc gcaaggctaa 840
aactcaaagg aattgacggg ggcccgcaca agcggcggag catgtggctt aattcgacgc 900
aacgcgaaga accttaccaa ggcttgacat acaccggaaa cgtccagaga tgggcgcccc 960
cttgtggtcg gtgtacaggt ggtgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg 1020
ttaagtcccg caacgagcgc aacccttgtc ccgtgttgcc agcaggccct tgtggtgctg 1080
gggactcacg ggagaccgcc ggggtcaact cggaggaagg tggggacgac gtcaagtcat 1140
catgcccctt atgtcttggg ctgcacacgt gctacaatgg ccggtacaat gagctgcgat 1200
accgcgaggt ggagcgaatc tcaaaaagcc ggtctcagtt cggattgggg tctgcaactc 1260
gaccccatga agtcggagtc gctagtaatc gcagatcagc attgctgcgg tgaatacgtt 1320
cccgggcctt gtacacaccg cccgtcacgt cacgaaagtc ggtaacaccc gaagccggtg 1380
gcccaacccc cttgtgggga gggagctgtc gaaggtgat 1419
Claims (2)
1. limestone efficiently corrodes actinomycetes, its Classification And Nomenclature be thermophilic carbon monoxide streptomycete (
streptomyces thermocarboxydus) NL-1, be preserved in China typical culture collection center, deposit number CCTCC NO:M 2012460, preservation date: on November 19th, 2012; Preservation address: Wuhan, China Wuhan University.
2. limestone according to claim 1 efficiently corrodes actinomycetes and is corroding the application in rock; Described rock is lime rock.
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| CN201310018611.4A CN103103151B (en) | 2013-01-18 | 2013-01-18 | Streptomyces thermocarboxydus NL-1 as actinomycete capable of efficiently eroding limestone and application of Streptomyces thermocarboxydus NL-1 |
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| CN102061270A (en) * | 2010-04-20 | 2011-05-18 | 怀化学院 | Streptomyces thermocarboxydus strain and cultivating method thereof |
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| CN102061270A (en) * | 2010-04-20 | 2011-05-18 | 怀化学院 | Streptomyces thermocarboxydus strain and cultivating method thereof |
Non-Patent Citations (2)
| Title |
|---|
| "典型细菌、真菌、放线菌对石灰岩动态溶蚀效果比较";贾丽萍;《应用与环境生物学报》;20071231;第13卷(第1期);摘要，126页1.1节 * |
| "碳酸钙微生物风化试验研究";丁丽君;《中国岩溶》;20080930;第27卷(第3期);197-200页 * |
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