CN103333838A - Alterikurthia sp and its application - Google Patents
Alterikurthia sp and its application Download PDFInfo
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- CN103333838A CN103333838A CN2013102818299A CN201310281829A CN103333838A CN 103333838 A CN103333838 A CN 103333838A CN 2013102818299 A CN2013102818299 A CN 2013102818299A CN 201310281829 A CN201310281829 A CN 201310281829A CN 103333838 A CN103333838 A CN 103333838A
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- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 235000010958 polyglycerol polyricinoleate Nutrition 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
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- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- RULSWEULPANCDV-PIXUTMIVSA-N turanose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](C(=O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RULSWEULPANCDV-PIXUTMIVSA-N 0.000 description 1
- 235000019143 vitamin K2 Nutrition 0.000 description 1
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Images
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a strain of Alterikurthia sp and its application. The Alterikurthia sp is named as Alterikurthia sp BC-LY168. Analytical tests find that the strain has the activity of hydrolyzing phosphoric acid, hydrolyzing organic acid ester, hydrolyzing polysaccharides, resisting pathogenic fungus growth, and promoting plant growth in itself. The 168rRNA gene sequence of the strain is shown as SEQ ID NO.1. By analysis of its enzyme activity and carbon source utilization characteristics, seed germination bioassay, phytopathogen resistance test to verify the efficacy of the Alterikurthia sp, the Alterikurthia sp can be used for plant growth promotion, soil improvement or microbial fertilizers.
Description
Technical field
The present invention relates to microorganism, relate in particular to a strain and like to reach the special Salmonella of Niccol and application thereof.
Background technology
The farmland uses chemical fertilizer no doubt can provide the crop initial stage required nutrient fast in a large number, but can cause Tu AUTHOR acidifying, fertility inequality to be polluted with underground water source.In the known Tu AUTHOR microorganism if having nitrogen fixation, ineffectivity nutrition dissolving and decompose function organic among the Tu AUTHOR, can be for being microbial fertilizer, and microbial fertilizer can replace partly chemical fertilizer, to solve the derived problem of unearthed AUTHOR deterioration of long-term application chemical fertilizer.Plant-growth promotes root circle bacterium (plant growth promoting rhizobacteria; PGPR) have four according to its useful mechanism of action: (1) bio-feritlizer effect (biofertilization), (2) biological control (bio-control) effect, (3) biology are educated effect (biorcmediation), (4) plant hormone effect (phytostimulation) again.
At present the patent of passing through as biological control or the application of anti-Plant diseases purposes with microorganism includes genus bacillus (Bacillus sp.), series bacillus (Paenibacillus), burkholderia (Burkholderia sp.), pseudomonas (Pseudomonas sp.), Serratia (Serratia sp.), helicobacter pylori (Helicobacter sp.), yeast strain (Pseudozyma), streptomycete (Streptomycetes sp.), Trichoderma (Trichoderma), black-koji mould Pseudomonas such as (Aspergillus sp.), but do not comprise the Pseudomonas of bacterial strain of the present invention.
Summary of the invention
One of purpose of the present invention provides a strain and likes to reach Nico Salmonella BC-LY168, it is the bacterial strain of choosing behind cultivation, separation and the purifying of substratum by among the Wu Ran Tu AUTHOR of Taiwan, show the most similar to Cole Te Shi (Kurthia) bacterium through the strain identification result, therefore called after likes to reach Nico Salmonella (Alterikurthia sp) BC-LY168, oneself is stored in the Chinese typical culture collection center that is positioned at Wuhan this bacterium on April 3rd, 2013, preserve to be numbered CCTCC NO:M2013118.
Two of purpose of the present invention provides the preparation of a kind of Ai Dani Al Kut Salmonella (Alterikurthia) of novelty, and its main cording has (1) phosphohydrolase: comprise alkaline phosphatase lytic enzyme (alkaline phosphatase), acid phosphatase lytic enzyme (acid phosphatase) and naphthols-AS-B1-phosphohydrolase (naphthol-AS-B1-phosphate hydrolase); (2) organic acid acetic lytic enzyme: comprise butyric acid esterase (butyrate esterase), sad esterase (caprylate esterase); (3) carbohydrate lytic enzyme: comprise alpha-glucosidase (α-glucosidase) and amylolytic enzyme (amylase); (4) growth for pathogenic fungi has antagonistic ability; (5) ability of promotion plant-growth, the bacterial classification of at least 5 kinds of functions can have organic matter decomposition for exploitation, promote plant-growth and the microbial preparation that Plant diseases is had the biological control function.
In order to achieve the above object, the present invention has adopted following technical scheme: a kind of love reaches the special Salmonella (Alterikurthia sp) of Niccol, and called after likes to reach the special Salmonella BC-LY168 of Niccol, and its deposit number is CCTCC NO:M2013118.
Described love reaches the 16S rRNA sequence of the special Salmonella of Niccol shown in SEQ ID NO.1.
Above-mentioned love reaches the special Salmonella of Niccol through the resulting cultivation of pure culture bacterium liquid.
Above-mentioned love reaches the special Salmonella of Niccol through pure culture and the outer supernatant liquor of born of the same parents centrifugal or that filtration obtains, and it comprises the outer metabolic active substance of thalline.
Above-mentioned love reaches the special Salmonella of Niccol through pure culture, centrifugal or filter resulting viable bacteria body.
Comprise by above-mentioned love and reach the special Salmonella of Niccol through the resulting cultivation of pure culture bacterium liquid, the outer supernatant liquor of born of the same parents or and the composition of viable bacteria body.
The application of above-mentioned composition is to be infected by plant pathogenic fungi for the control crop, or is used for promoting the Chang And Gai of Zhi thing Sheng Liang Tu AUTHOR quality.
Described plant pathogenic fungi refers to fusarium (Fusarium) fungi.
The described method that is infected by plant pathogenic fungi for the control crop is that described composition is contacted with plant seed, fruit or root.
Described composition also comprises plant-growth and promotes microorganism, organic fertilizer or and agrochemical substances.
Test by analysis, love of the present invention reach the special Salmonella BC-LY168 of Niccol itself and have the growth of hydrolysis phosphoric acid, hydrolysis organic acid acetic, hydrolysis Polysaccharides, antagonism pathogenic fungi, the ability activity of promotion plant-growth.The characteristic of its enzyme activity, utilization of carbon source, seed germination bioanalysis, phytopathogen resistance test to verify its effect by analysis, can be used for promoting the purposes of plant-growth, Tu AUTHOR improvement or microbial fertilizer.
Description of drawings
Fig. 1 approaches the close source dendrogram of the bacterial classification 16S rRNA gene order of relation for utilizing Kimura-2parameter pattern Kimura-double factor to analyze BC-LY168T (deletion) with other.
Fig. 2 is with the antagonistic action of potato hexose nutrient agar (PDA) test bacterial strain BC-LY168 of the present invention for phytopathogen Fusarium oxysporum growth, wherein, (A) phytopathogen Fusarium oxysporum f.sp.niveum (E.F.Smith) Snyder ﹠amp; Hanson (FNH0103); (B) FNH0103 bacterial strain+BC-LY168 bacterial strain; (C) Fusarium oxysporum f.sp.conglutinans (FOC19); (D) FOC bacterial strain+BC-LY168 bacterial strain.
Fig. 3 comprises Fusarium oxysporum f.sp.niveum (E.F.Smith) Snyder ﹠amp for BC-LY168 for phytopathogen Fusarium oxysporum; The growth-inhibiting per-cent of Hanson (FNH0103), Fusarium oxysporum f.sp.conglutinans (FOC19) and Fusarum oxysporum f.sp.cubense race4 (FOC24) bacterial strain.
Embodiment
The fundamental characteristics of bacterial strain BC-LY168 of the present invention is as follows:
1, the feature of thalli morphology
This bacterial strain is gram-positive microorganism, and the tool movability is the aerobic bacterium; In trypsinase salt agar glue (Tryptic soy agar, TSA) or nutrition agar glue (Nutrient agar, NA) substratum when growth colony shape is class root shape.
2, microbial culture character
The growth temperature of bacterial strain of the present invention is 30-37 ℃, the different battalion's property of 0-10%NaCl chemistry, and growth pH value is 5.0-10.0, is known in nutrient agar (Hi-Media), tryptone soy agar (BBL) and R2A (oxoid) substratum can be grown.
3, the biochemical characteristic of bacterial strain
Bacterial strain of the present invention has following enzyme activity: catalase (catalase), alkaline phosphatase lytic enzyme (alkaline phosphatase), acid phosphatase lytic enzyme (acid phosphatase), naphthols-AS-B1-phosphorus lytic enzyme (naphthol-AS-B1-phosphohydrolase), butyric acid esterase (butyrate esterase), sad esterase (caprylate esterase), alpha-glucosidase (α-glucosidase) and amylolytic enzyme (amylase).
4, the physiological property of bacterial strain
The carbon source that bacterial strain of the present invention can utilize: 2, the 3-butyleneglycol (2,3-butanediol), cyclohexaamylose (α-cyclodetxrin), D-galactose aldehydic acid (D-gacturonic acid), γ-hydroxybutyric acid (γ-hydroxybutyric acid), phenylethylamine (phenylethylamine), α-ketone valeric acid (α-keto valeric acid), left-handed-tyramine acid (L-threonine), interfacial agent (Tween-40, Tween-80) and dextrorotation-trehalose (D-trehalose).Can be produced the carbohydrate of acid by bacterial strain utilization of the present invention: glycerine (glycerol), dextrorotation-ribose (D-ribose), dextrorotation-sorbitol (D-sorbitol), dextrorotation-maltose (D-maltose), dextrorotation-turanose (D-turanose), gluconic acid (Gluconate), potassium-5-ketone group gluconic acid (potassium-ketogluconate).
The composition of known plants root exudates generally include carbohydrate, aminoacids, vitamins, organic acid and other various kinds of compounds, conducive to the use of microorganism, is considered to be active main reason of micro organism quantity and increased activity.Therefore bacterium also is considered to successfully one of the factor of planting a colony root circle of bacterium to the ability of utilizing of various carbon sources.
The present invention's love reaches special Salmonella (Alterikurthia sp) BC-LY168 of Niccol and includes following feature:
One, bacterial strain of the present invention is the natural wild-type strain of automatic pollution soil AUTHOR environment separation, without the processing of any artificial genetic modification.
Two, bacterial classification of the present invention is a novel strain, possesses the bacterial classification that decomposes organic matter, bio-feritlizer and biological control.
Based on above-mentioned feature, this bacterium can be developed as microbial preparation.
The present invention's bacterial strain can be done following application: (1) microbial fertilizer, can be applied to the root circle, and promote plant-growth; (2) Tu AUTHOR modifying agents are applied to Tu AUTHOR, and (3) compost or organic quality-improving are applied to the compost that do not become thoroughly decomposed or become thoroughly decomposed or organic matter, (4) have the purposes of the rhizosphere microorganism Inoculant of biological control function for specific fungal disease among the Tu AUTHOR.
According to the present invention, can use the constituent processing of liking to reach the special Salmonella BC-LY168 of Niccol.The present invention's specific embodiment is for liking that reaching the special Salmonella BC-LY168 of Niccol yeast culture proves that in solid medium its viable bacteria body itself has the effect that suppresses the Fusarium phytopathogen.
Now described in detail the present invention with following example, only And does not mean that the present invention only is confined to these examples and holds within disclosing.
[separation screening of Taiwan sheath amine alcohol unit cell]
From the soybean fermented sample of gathering from Taiwan Province, to get the 1g sample and put into the 10ml sterilized water, dilution is applied in nutrient agar medium (nutrient agar) (HIMEDIA) on the substratum, in 30 ℃ of cultivations, separates and the bacterial strain of the single bacterium colony of purifying.
[analyzing the present invention's BC-LY168 bacterial strain with multinomial sorting technique]
1. genotypic classification analysis
Identify the present invention's bacterial strain with 16S rRNA gene sequencing, it is that the microbial staining body DNA separation sleeve group (UltraCleanTM Microbial Genomic DNA Isolation Kit) of utilizing MO BIO company (USA) to provide is carried out the DNA extraction, sub-cell (small-subunit with intestinal bacteria (Escherichia coli), SSU) the height reservation queue design bacterium specificity primer (bacterial universal primer) of nuclear candy body rna gene, forward primer 1F (5 '-GAG TIT GAT CAT GGC TCA G-3 ') and reverse primer 9R (5 '-AAG G AG GTG ATC CAA CCG CA-3 ') (as shown in table 1) carry out polymerase chain reaction (PCR), add bacterial chromosome DNA Approximately 20-50ng in the PCR reaction solution respectively as masterplate, two gangs of each 20pmol of primer, 1.5U Taq archaeal dna polymerase, 200 μ M dNTPs, it is 25 μ L that buffered soln makes cumulative volume, reaction conditions be 95 ℃ 5 minutes, 95 ℃ 1 minute, 50 ℃ 1 minute 30 seconds, 72 ℃ 2 minutes, totally 35 the circulation, 72 ℃ 7 minutes.Carry out the propagation of 16S rDNA with polymerase chain reaction.After recycling QIAquick Gel Extraction cover group reclaims DNA, desire the thermal cycle reaction of sequencing gene fragment, the sequencing primer of selecting for use is the 16S rDNA specificity primer of reservations highly, will carry out stain termination rotational ordering (dye terminator cycle sequencing) with sequencing with primer through the fragment of bacterium specificity primer amplification and react.At last with
310PRISM sequencing instrument carries out detecting and the sequence interpretation of fluorescent, obtain the 16SrDNA1 of bacterium BC-LY168 of the present invention, 447 base pairs (shown in SEQ IDNO.1), the 16S rDNA sequence of bacterial strain of the present invention have sent to deposit and have given birth to skill money gene information storehouse, News center (NCBI Genbank) to American National to obtain logining number (accession number) be JF311905.
The primer (primers) that table 1. is adopted with polymerase chain reaction (PCR) increment 16S rRNA
*: nucleic acid Wei Zhi Department refers to the position of the 16S rRNA nucleotide sequence of intestinal bacteria
The reference culture (type strains) that the Al Kut Salmonella (Kurthia) that NCBI GeneBank is obtained belongs to utilizes Clustal_X (1.83) formula to carry out parallelism, again with Mega-4 software (Tamura et al., 2007) carry out relationship analysis (phylogenetic analysis), utilize contiguous method of attachment (neighbour-joining method) (Saitou and Nei, 1987) inference to develop and set (evolutionary trees).(Felsenstein, 1985) are assessed in the opening up of tree of developing the at last bootstrapping analysis of learning with based on the contiguous connection method (neighbour-joining method) of 1000 stochastic analyses (re-sampling) (bootstrap analyses) of falling forward.Fig. 1 is the dendrogram constructed with proximity junction legal (neighbor-joining method) of bacterial strain BC-LY168 of the present invention for this reason, branch represents the percentage that still is attributed to same association after 1000 stochastic analyses with display digit, shows that bacterial strain BC-LY168 of the present invention and pseudomonas have dependency most.The similarity that shows bacterial strain of the present invention and Ji Shi Al Kut Salmonella (Kurthia gibsonii) NCIMB9758T (97.4%) is nearest; Be lower than 97% with the Xiang Si Du The of other bacterial classifications.Be not higher than 60% (Stackebrandt and Goebel, 1994) when the similarity of 16S rDNA is lower than 97% degree that is equivalent to the DNA-DNA heterozygosis, can assert it is novel species.Bacterial strain BC-LY168 of the present invention and Er Te Bordetella (Kurthia), Bill Rammell genus bacillus (Rummeliibacillus), green genus bacillus (Viridibacillus) and be respectively 3.2%, 3.6% and 4.0% from the distance of the evolution between the standard bacterial classification (type species) of amino acid genus bacillus (Lysinibacillus) (evolutionary distances), all the overgauge bacterial classification with belong in bacterial classification evolutions apart from (be respectively 1.3-2.8%, 1.9% and 0.1-2.8%), prove that bacterial strain BC-LY168 of the present invention is a novel species bacterium.
[the thalli morphology analysis of thalline]
Amending method according to Cowan (1974) will carry out gram (Gram) dyeing, the positive bacterium of bacterial strain gramstaining of the present invention.The thalline of bacterium of the present invention was grown 3 days down at 30 ℃ in movability liquid nutrient medium (mobility), with its movability of microscopic examination (mobility).The thalline kenel of utilizing transmission electron microscope to observe bacterial strain BC-LY168 of the present invention presents shaft-like, does not have spore with bacterium of the present invention, aerobic organism, and the cell when the growth logarithmic phase is shaft-like, has flagellum, so belong to movability.In nutrient agar medium (Nutrient agar; NA) the substratum grown cultures after 2 days the colony shape outward appearance be class root shape.
[physiology of thalline and the analysis of cultural property]
Inoculation of the present invention is tested the growth situation of following condition in Nutrient broth (Hi-Media) substratum:
Growth temperature
With the NB liquid nutrient medium of fresh inoculation in the different pH-values of adjusting with sodium hydroxide or hydrochloric acid (pH), test the pH that it can be grown under 30 ℃ of shaking culture, the result shows that its pH value scope that can grow is 5-10.
Growth situation under the NaCl of different concns, temperature are between 25-37 ℃ that chemical different battalion's property can be grown in containing the NB of 0-10.0%NaCl (HIMEDIA).
[thalline chemistry analysis]
1. cell fatty acid compositional analysis
Test inoculation likes to reach the kenel (profile) of cell walls lipid acid of other bacterial classifications of Niccol Salmonella BC-LY168 and Al Kut Bordetella, with bacterial strain in the Erlenmeyer flask that contains Nutrient agar substratum, measure its chemical classification character after 48 hours in 30 ℃ of cultivations, according to carrying out saponification, methylate and extract as Miller (1982), as described in Paisley (1996), cooperate microbial identification system (MIDI) to identify lipid acid with gas-flame ion detector (GC-FID) thalline.The cellular fat acid assay shows that the main lipid acid of BC-LY168 is anteiso-C15:0 (34%), iso-C15:0 (21%), anteiso-C17:0 (10%), iso-C6:0 (7%), n-C16:0 (6%), iso-C14:0 (6%) and C15:1 ω 5c (5%).The typical lipid acid of the special Bordetella of Cole (Kurthia) is iso-C14:0, iso-C15:0, anteiso-C15:0, C15:1 ω 5c, the main lipid acid that shows the main lipid acid composition Anteiso-C17:0 And Fei Keerte Bordetella typical case bacterial classification of bacterial strain BC-LY168 of the present invention, show that the BC-LY168 bacterial strain is that a new Shu ， And and called after like to reach the special Salmonella (Alterikurthia) of Niccol.
2. breathe the analysis of quinone
The isoprenoid (isoprenoid) of bacterial strain of the present invention being liked to reach the special Salmonella BC-LY168 of Niccol extracts with people's such as Minnikin (1984) method, (HPLC) analyzes (Collins with high performance liquid chromatography, 1985), the result shows: bacterial strain of the present invention has the insatiable hunger of 7 isoprene units and closes vitamin k4 (menaquinone; MK-7).
[analyzing the utilization of carbon source ability of BC-LY168 bacterial strain with BIOLOG]
Love is reached the setting-out of the special Salmonella BC-LY168 of Niccol bacterial strain be seeded to NA culture medium culturing ware, cultivated 16-24 hour in 30 ℃, bacterium colony is suspended in the GN/GP-IF Inoculant, adjust the transparence 65% of bacterial concentration to the wavelength 590nm, getting 150 μ l adjustment suspension annotates to the micro-square position of GN2BIOLOG96 hole slot (wells), place 30 ℃ to cultivate 72 hours, measure the light absorption value of its wavelength 595nm with BIOLOG Thermomax interpretoscope, interpretation via interpretoscope, each can be cultivated OD value (the λ 1=590 of the color reaction of the different depths in the well, λ 2=750) deducts A-1 and cultivate well (blank test, carbonaceous sources not) numerical value can be divided into strong reaction, the weak reaction and reactionless three kinds.Learnt by the BIOLOG analytical results, the available carbon source of BC-LY168 bacterial strain of the present invention's separation has 2, the 3-butyleneglycol (2,3-butanediol), cyclohexaamylose (α-cyclodetxrin), D-galactose aldehydic acid (D-gacturonic acid), γ-hydroxybutyric acid (γ-hydroxybutyric acid), phenylethylamine (phenylethylamine), α-ketone valeric acid (α-keto valeric acid), left-handed-tyramine acid (L-threonine), interfacial agent (Tween-40, Tween-80) and dextrorotation-trehalose (D-trehalose).Known, the secretion of plant roots circle contains the volume carbohydrate and the carbonyl acid，anticipatedly applied to the plant roots circle with bacterial strain of the present invention, and it can have root to plant a colony the advantage of effect, for plant, can bring into play the physiological function that bacterium itself has.
[measuring the physiological property of BC-LY168 bacterial strain with API20E, API20NE, API-ZYM cover group]
1. oxydase and catalase activity test:
Test its oxidase activity according to the specification sheets that bioMerieux manufacturer provides, bacterium colony is directly dripped with oxidase reagent (OX reagent), being purple is positive reaction (positive reaction), represent that bacterium BC-LY168 bacterial strain of the present invention can secrete oxydase (oxidase), drip with 3% hydrogen peroxide (hydrogen peroxide) and on bacterium colony, can produce bubble, represent that bacterial strain of the present invention can secrete catalase (catalase).
2.API20NE, API ZYM test
With the biochemical activity of API20NE test bacterial strain of the present invention, as the operation instructions of cover group (bioM é rieux, France).Its step is as described below: bacterium is lined on the solid medium that is fit to growth, in 30 ℃ of cultivations.Pure bacterium colony is added the 2ml normal saline solution, mix, A600 is adjusted into 0.5 with the thallus suspension liquid light absorption value.Sterilized water is filled groove in culture plate, to keep the moisture of culture plate.API20NE is reacted bar (strip) place culture plate.Thallus suspension liquid is splashed among the tube of NO3 to PNPG test holes.The cupule of GLU, ADH, URE test holes covers sterile mineral oil.The thallus suspension liquid of getting 200 μ l adds among the AUX medium, the composition of substratum is divided into: 2g/L ammonium sulfate, 10.5ml/L VITAMIN solution, 10ml/L trace element (trace element), 6.24g/L Lin Suan Na (monosodium phosphate), 1.5g/L Repone K (potassium chloride), 1.5g/L agar (agar), pH7.0-7.2, evenly mixing.The thallus suspension liquid of allocating is added among the tube and cupule of GLU to PAC12 test holes.After the culture plate lid covered, cultivated 48 hours in 30 ℃.Add each one of NIT1 and NIT2 reagent to the NO3 test holes, react interpretation after 5 minutes.Add a JAMES reagent in the TRP test holes, interpretation immediately.Single-minded enzyme activity with API-ZYM cover group (bioMerieux, Inc. company) test bacterial strain BC-LY168 bacterial strain of the present invention.After at first getting bacterium colony on the culture dish with transfering loop, be dissolved in the 5ml sterilized water, make transparence near 50% through adjusting bacterial concentration, get the bacterium liquid 65 μ l that adjust add each the reaction hole slot in after, sample is placed 30 ℃ of reactions 24 hours, adding each Di ， Static of API-ZYM cover group A liquid and B liquid again puts after 5 minutes and observes colour-change.By API20NE and API ZYM test result, learn that the BC-LY168 bacterial strain has the activity of following ferment: alkaline phosphatase lytic enzyme (alkaline phosphatase), acid phosphatase lytic enzyme (acid phosphatase), naphthols-AS-B1-phosphorus lytic enzyme (naphthol-AS-B1-phosphohydrolase), butyric acid esterase (butyrate esterase), sad esterase (caprylate esterase), alpha-glucosidase (α-glucosidase) and amylolytic enzyme (amylase).
[with the antagonism analysis of bacterium at phytopathogen]
Selection has pathogenic three fungal strain bacterial classification Fusarium oxysporums (Fusarium oxysporum) for plant and is respectively watermelon dead arm pathogenic bacterium Fusarium oxysporum f.sp.niveum (E.F.Smith) Snyder﹠amp; Hanson (FNH0103), broccoli yellowtop pathogenic bacterium Fusarium oxysporum f.sp.conglutinans (FOC19), with banana yellowtop pathogenic bacterium Fusarum oxysporum f.sp.cubense race4 (FOC24), wherein FNH0103 and FOC19 two bacterial strains system to plant disease by Taiwan Chung Hsing University be that the Huang culture and education of shaking is awarded institute and provided, FOC24 bacterium strain The is that to plant disease by Taiwan Chung Hsing University be that associate professor Zhang Bifang is provided.
With bacterial strain streak inoculation of the present invention in nutrient agar (NA, Hi-Media) go up in 30 ℃ of cultivations after 1 day, the bacterium colony of growing is suspended in no Jun Shui And adjusts its light absorption value A600=0.5, get 100 μ l thallus suspension liquids to potato hexose agar (Potato dextrose agar; PDA) substratum, smear evenly after, cut in the middle of the PDA of cultivation based on the top BC-LY168 of scribbling thalline that a square (0.42cm) contains true bacterium bacterium Silk from growing in the PDA substratum again.In 25 ℃ of incubators (top is to shine 12 hours the every day of 20W fluorescent lamp 18-inch distance) cultured continuously 6 days.The size of observed and recorded fungal growth.The result shows that the BC-LY168 bacterial strain can obviously suppress Fusarium oxysporum bacterial strain in the growth of PDA substratum as shown in Figures 2 and 3.
Sequence table
＜110〉go up company limited of the luxuriant biotechnology of Heiden research and development centre
＜120〉strain likes to reach the special Salmonella of Niccol and application thereof
<160>1
<210>1
<211>1447
<212>DNA
＜213〉like to reach special Salmonella (Alterikurthia sp) BC-LY168 of Niccol
<400>1
GAGTTTGATC ATGGCTCAGA ACGAACGCTG GCGGCGTGCC TAATACATGC AAGTCGAGCG 60
AATGACGAGA AGGTTGCTTC TCTGATTTAG CGGCGGACGG GTGAGTAACA CGTGGGCAAC 120
CTGCCCTGTA GACTGGGATA ACTTCGGGAA ACCGGAGCTA ATACCGGATA ATTCTTTTAG 180
CCTCATGGCT TTAAGCTAAA AGGCGCTTCG GCGTCACTAC AGGATGGGCC CGCGGTGCAT 240
TAGCTAGTTG GTGCGGTAAC GGCCTACCAA GGCAACGATG CATAGCCGAC CTGAGAGGGT 300
GATCGGCCAC ATTGGGACTG AGACACGGCC CAAACTCCTA CGGGAGGCAG CAGTAGGGAA 360
TCTTCCACAA TGGACGAAAG TCTGATGGAG CAACGCCGCG TGAGTGATGA AGGTTTTCGG 420
ATCGTAAAAC TCTGTTGTAA GGGAAGAACA AGTGCGTTAG GTAATGAACG CACCTTGACG 480
GTACCTTATT AGAAAGCCAC GGCTAACTAC GTGCCAGCAG CCGCGGTAAT ACGTAGGTGG 540
CAAGCGTTGT CCGGATTTAT TGGGCGTAAA GCGCGCGCAG GTGGTTTCTT AAGTCTGATG 600
TGAAAGCCCA CGGCTCAACC GTGGAGGGTC ATTGGAAACT GGGAGACTTG AGTGCAGAAG 660
AGGATAGTGG AATTCCAAGT GTAGCGGTGA AATGCGTAGA GATTTGGAGG AACACCAGTG 720
GCGAAGGCGA CTGTCTGGTC TGTAACTGAC ACTGAGGCGC GAAAGCGTGG GGAGCAAACA 780
GGATTAGATA CCCTGGTAGT CCACGCCGTA AACGATGAGT GCTAAGTGTT AGGGGGTTTC 840
CGCCCCTTAG TGCTGCAGCT AACGCATTAA GCACTCCGCC TGGGGAGTAC GACCGCAAGG 900
TTGAAACTCA AAGGAATTGA CGGGGGCCCG CACAAGCGGT GGAGCATGTG GTTTAATTCG 960
AAGCAACGCG AAGAACCTTA CCAGGTCTTG ACATCCCAAT GACCGTCTTA GAGATAAGAT 1020
TTTCCCTCCG GGGACATTGG TGACAGGTGG TGCATGGTTG TCGTCAGCTC GTGTCGTGAG 1080
ATGTTGGGTT AAGTCCCGCA ACGAGCGCAA CCCTTATTCT TAGTTGCCAT CATTTAGTTG 1140
GGCACTCTAA GGAGACTGCC GGTGACAAAC CGGAGGAAGG TGGGGATGAC GTCAAATCAT 1200
CATGCCCCTT ATGACCTGGG CTACACACGT GCTACAATGG ACGGTACAAA GAGCTGCAAG 1260
CCCGCGAGGG TTAGCCAATC TCATAAAACC GTTCTCAGTT CGGATTGTAG TCTGCAACTC 1320
GACTACATGA AGCCGGAATC GCTAGTAATC GCGGATCAGC ATGCCGCGGT GAATACGTTC 1380
CCGGGCCTTG TACACACCGC CCGTCACACC ACGAGAGTTT GTAACACCCG AAGCCGGTGG 1440
GGTAACC 1447
Claims (10)
1. a strain likes to reach the special Salmonella (Alterikurthia sp) of Niccol, and called after likes to reach the special Salmonella BC-LY168 of Niccol, and its deposit number is CCTCC NO:M2013118.
2. love as claimed in claim 1 reaches the special Salmonella of Niccol, it is characterized in that: described love reaches the 16S rRNA sequence of the special Salmonella of Niccol shown in SEQ ID NO.1.
3. reach the special Salmonella of Niccol through the resulting cultivation of pure culture bacterium liquid by the described love of claim 1.
4. reach the special Salmonella of Niccol through pure culture and the outer supernatant liquor of born of the same parents centrifugal or that filtration obtains by the described love of claim 1, it comprises the outer metabolic active substance of thalline.
5. reach the special Salmonella of Niccol through pure culture, centrifugal or filter resulting viable bacteria body by the described love of claim 1.
6. comprise by the described love of claim 1 and reach the special Salmonella of Niccol through the resulting cultivation of pure culture bacterium liquid, the outer supernatant liquor of born of the same parents or and the composition of viable bacteria body.
7. the application of the described composition of claim 6 is characterized in that: be used for the control crop and infected by plant pathogenic fungi, or be used for promoting the Chang And Gai of Zhi thing Sheng Liang Tu AUTHOR quality.
8. application as claimed in claim 7 is characterized in that: described plant pathogenic fungi refers to fusarium (Fusarium) fungi.
9. application as claimed in claim 7 is characterized in that: the described method that is infected by plant pathogenic fungi for the control crop is that described composition is contacted with plant seed, fruit or root.
10. composition as claimed in claim 6 is characterized in that: described composition also comprises plant-growth and promotes microorganism, organic fertilizer or and agrochemical substances.
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| CN116286449A (en) * | 2022-10-28 | 2023-06-23 | 云南大学 | Bacillus viridis YSL-1-5 capable of promoting growth of armillaria mellea and application thereof |
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| RU2270063C2 (en) * | 2003-03-07 | 2006-02-20 | Ооо Пкф "Бигор" | Bacterium kurthia zopfii strain capable to recover oil, petroleum products and aromatic polycyclic hydrocarbons |
| CN102433289A (en) * | 2012-01-16 | 2012-05-02 | 江南大学 | Strain for producing citrulline and method for biologically synthesizing citrulline with same |
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| RU2270063C2 (en) * | 2003-03-07 | 2006-02-20 | Ооо Пкф "Бигор" | Bacterium kurthia zopfii strain capable to recover oil, petroleum products and aromatic polycyclic hydrocarbons |
| CN102433289A (en) * | 2012-01-16 | 2012-05-02 | 江南大学 | Strain for producing citrulline and method for biologically synthesizing citrulline with same |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116286449A (en) * | 2022-10-28 | 2023-06-23 | 云南大学 | Bacillus viridis YSL-1-5 capable of promoting growth of armillaria mellea and application thereof |
| CN116286449B (en) * | 2022-10-28 | 2024-04-16 | 云南大学 | Bacillus viridis YSL-1-5 capable of promoting growth of armillaria mellea and application thereof |
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