CN101012444A - Rahnella aquatilis HX2 and application thereof - Google Patents
Rahnella aquatilis HX2 and application thereof Download PDFInfo
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Abstract
The invention discloses a new Rahnella aquatilis strain HX2 (CGMCC No.1896), which is characterized by the following: manufacturing antagonistic material and IAA; inhibiting partial pathogenic bacteria (Agrobacterium, Xanthomonas, Clavibacter, Pseudomonas and Bacillus) and partial pathogenic fungus (Fusarium, Alternaria and Botrytis); detecting root cancer based on green house sunflower as indicating plant.
Description
Technical field
The present invention relates to new bacterial strain of a kind of microorganism and application thereof, specifically Rahnella aquatilis HX 2 and the application in biological control thereof.
Background technology
The Root of European Grape carninomatosis generally takes place in the world.The classical symptom of Root of European Grape carninomatosis is to form tumour.Site of pathological change mainly at trunk near the position (collar portion) on ground, also can take place on place or the overhead nearer grape trellis face slightly down at the soil table.The Root of European Grape carninomatosis all has generation in various degree in the orchard of many provinces, cities and regions of northeast, North China, northwest, the Yellow River and the Yangtze valley of China and nursery, wherein ground diseases such as Liaoning, Jilin, the Inner Mongol, Beijing, Hebei, Shanxi, Shandong, Zhejiang and Shanghai take place serious.Some vineyard sickness rate are up to 80-90%, and production loss reaches 30-70%, and the when serious even garden of ruining has no harvest, and causes heavy economic losses.Because the generation of root knot is relevant with the Ti-plasmids of pathogenic bacteria, behind edaphic bacillus invaded plants wounded tissue, T-DNA on its Ti-plasmids can enter vegetable cell and be incorporated on the DNA of plants, T-DNA causes the formation of crown gall in the intravital expression of plant, so uses general chemical agent and can not obtain good prevention effect.Nineteen seventies, Australian Kerr found the K84 bacterial strain, made the control of root knot important breakthrough occur, and after this, the application biocontrol strain carries out biological control to root knot and obtained remarkable effect.But K84 is only effective to the A.tumefaciens and the A.rhizogenes crown gall bacterium that contain nopaline type or agrobacteriocin A type Ti-plasmids, does not imitate anti-to the Root of European Grape carninomatosis that A.vitis causes.Therefore, the biocontrol strain of seeking multiple source becomes the main means of control Root of European Grape carninomatosis.At present, the report of Agrobacterium, the Pseudomonas of existing many strains no pathogenicity and Bacillus control Root of European Grape carninomatosis.Bell report Rahnella aquatili JC577 bacterial strain has dull and stereotyped restraining effect to the grape edaphic bacillus, yet it can not prevent and treat the Root of European Grape carninomatosis.
Summary of the invention
The object of the present invention is to provide new bacterial strain HX2 of a kind of aquatic engler of drawing bacterium and the application in biological control thereof.
Bacterial strain HX2 of the present invention separates the new bacterial strain of the aquatic engler of the drawing bacterium (Rahnella aquatilis) that obtains from the soil of vineyard, outer suburbs, Beijing, China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC), preserving number CGMCC No.1896 have been preserved in on December 22nd, 2006.
HX2 specifically separates by the following method and obtains: from vineyard, outer suburbs, Beijing, gather soil sample, get 10g add in the triangular flask of dress 90mL stroke-physiological saline solution, 28 ℃, 150rpm shaking table concussion 15min, after leaving standstill 10min, get 0.5mL and add in the 4.5mL stroke-physiological saline solution, dilute 10 successively again
-2, 10
-3, 10
-4Doubly, get above soil suspension 100 μ L respectively at NA substratum (extractum carnis 5g/L, peptone 10g/L, NaCl 5g/L, agar 15g/L, pH7.0-7.2) even coated plate on the flat board dries up in the super quiet worktable, and each concentration repeats 3 times, cultivate 36h in 28 ℃ of incubators, behind the single bacterium colony line of aseptic toothpick picking bacterium purifying, K308 is the target bacterium with grape edaphic bacillus (A.vitis), utilizes double-deck culture method to carry out antimicrobial preliminary screening short of money; To detecting the greenhouse protection effect behind the antagonism bacterium shake flask fermentation that obtains, finishing screen is selected the aquatic engler of the drawing bacterium of the good biocontrol strain of proterties (Rahnellaaquatilis) HX2.
HX2 has following characteristic:
Gram-negative, electron microscope observation, thalline rod-short, diameter are 0.8 μ m, many flagellums of Zhousheng (see figure 1).Can grow containing on the substratum of 2%NaCl, can under 4 ℃～41 ℃ culture condition, grow, reaction such as catalase, Citrate trianion utilization, VP, arginine dihydrolase, malonate utilization, phenylalanine utilization is positive, glucose fermentation produces sour aerogenesis, fermentation pectinose, sorbyl alcohol, sucrose produce acid, oxydase, methyl red, phenylalanine deaminase, lysine decarboxylase, gelatine liquefication, hydrocellulose, amylase, H
2Reaction negatives such as S, tartrate utilization, acetate utilization.
(the GENBANK accession number: U88436.1) similarity is the highest, reaches 99% with engler bacterium (CDC2987-79) the 16S rDNA that draws that delivers at present for the 16S rDNA sequence of HX2 of the present invention.Comprehensive morphological feature and 16S rDNA Sequence Identification HX2 are the aquatic new bacterial strain of engler bacterium (Rahnella aquatilis) that draws.
Studies show that HX2 has good bacteriostatic action to the various plants pathogenic bacteria, has good preventive and therapeutic effect to the Root of European Grape carninomatosis.
The present invention also provides the fermentation culture method of HX2, comprises step:
1, actication of culture
Use improvement 523 substratum, culture medium prescription is: sucrose 5-10g/L, KH
2PO
41.0-2.0g/L, yeast extract paste 5-10g/L, CaCO
34.0-6g/L, NaCl 0.1-0.3g/L, MgSO
47H
2O 0.2-0.4g/L, agar 15-18g/L, surplus is a water, pH7.0-7.2.The aquatic engler of drawing bacterium (Rahnella aquatilis) HX2 is inoculated on improvement 523 medium slant, cultivated 20-24 hour for 25-28 ℃.
2, culture of seed liquid
Seed culture medium is formed: sucrose 4-6g/L, corn steep liquor 1-3g/L, KH
2PO
40.03-0.0.07g/L, K
2HPO
40.03-0.07g/L, MgSO
47H
2O 0.025-0.05g/L, (NH
4)
2SO
40.3-0.8g/L, CaCO
31-3g/L, surplus is a water, pH7.0-7.2.The HX2 that activation is good is mixed with 10 with stroke-physiological saline solution
8-10
9The bacteria suspension of CFU/mL is inoculated in the seed culture medium with the inoculum size of 1-2%, and 25-28 ℃ of shaking table concussion cultivated, and rotating speed is 130-180rpm, and incubation time is 12-16 hour.
3, ferment tank
Fermention medium consists of: glucose 10-15g/L, yeast extract paste 2-4g/L, NaCl0.5-0.8g/L, KH
2PO
40.25-0.5g/L, K
2HPO
40.25-0.5g/L, MgSO
47H
2O0.5-1g/L, (NH
4)
2SO
45-10g/L, rapeseed oil 0.4-0.6g/L, surplus is a water, pH7.0-7.2.Per minute air flow volume), tank pressure 0.05-0.06MPa cultured seed liquid is inserted in the fermentor tank with the inoculum size of 2-4%, and 20-28 ℃, stirring velocity is 150-180rpm, and air flow is 1: 0.6-0.8 (fermentating liquid volume:.Fermented 20-24 hour, the bacterium amount reaches 5-10 * 10
8CFU/mL obtains HX2 bacterium liquid.
Because HX2 can produce antimicrobial substance in a large number in the substratum that contains glucose and yeast extract paste, have bacteriostatic activity, so fermention medium to select glucose and yeast extract paste for use be carbon nitrogen source.
The present invention also provides the microbial inoculum that contains HX2, its can by above-mentioned fermented liquid and turfy soil are mixed must.
Bacterial strain of the present invention is used for biological control, can reduce the sickness rate of plant root cancer effectively, for the high yield of grape has been created condition.
Description of drawings
Fig. 1 is morphologic observation (* 10000) under the HX2 bacterial strain transmission electron microscope of the present invention.
Embodiment
Following embodiment is used for further specifying of the present invention, but is not used for limiting the scope of the invention.
Embodiment 1 HX2 bacterial strain 16S rDNA analyzes
With the aquatic engler of drawing bacterium (Rahnella aquatilis) HX2 genomic dna is template, with 16S amplification universal primer 63F 5 '-CAGGCCTAACACATGCAAGTC-3 ' and 1494R 5 '-GGYTACCTTGTTACGACTT-3 '.To increase in the following pcr amplification reaction system: 50 μ L amplification systems, 10pmol primer, 0.2mM dNTP, 1.5mMMgCl
2, 2.5 U Taq archaeal dna polymerases (TaKaRa), 1 * PCR buffer (TaKaRa), the 25ng genomic dna is made template.PCR reaction conditions: 94 ℃ of sex change 5min; 94 ℃ of sex change 40sec, 55 ℃ of renaturation 40sec, 72 ℃ are extended 1min, and cycle number is 35; 72 ℃ are extended 10min.The PCR product reclaims purifying with Omega " Gelextraction kit " behind 1% agarose gel electrophoresis, connect pMD18-T Vector, Transformed E .coli DH5 α, check order with ABI 3730 dna sequencing instrument in Invitrogen company, its nucleotide sequence is shown in sequence table SEQ ID NO.1.With blast program GenBank (
Http:// www.ncbi.nlm.nih.gov) result of the corresponding sequence comparison 16SrDNA sequence that shows HX2 with deliver at present draw engler bacterium similarity the highest, reach 99%.
Embodiment 2 antimicrobial spectrum analyses
To the dull and stereotyped detection method that suppresses of plant pathogenetic bacteria.Detect the inhibition situation of HX2 with double-deck culture method to plant pathogenetic bacteria.The activatory Rahnella aquatilis HX 2 is mixed with bacteria suspension (10
9CFU/mL), draw 10 μ L points on the PDA substratum, every ware was a bit cultivated 48 hours, and was killed the HX2 of growth with the 3mL trichloromethane with its steam for 28 ℃.After 10-12 hour, bacteria suspension is made on cultured edaphic bacillus bacterial strain and the former bacterium of bacterial blight of rice, cotton bacterial angular leaf spot pathogenic bacteria, cabbage black rot pathogenic bacteria, cucumber bacterial angular leaf spot pathogenic bacteria, canker of tomato pathogenic bacteria, subtilis inclined-plane respectively, determined that with Mike's turbidimetry whole bacteria suspension concentration is about 10
9CFU/mL.Draw 50 μ L bacteria suspensions respectively and add soft YEB substratum (sucrose 5g/L, peptone 5g/L, extractum carnis 5g/L, yeast extract paste 1g/L, agar 7g/L, the MgSO that 5mL melts postcooling to 50 ℃
47H
2O 0.5g/L, pH7.0-7.2) in, rapid mixing is poured on the substratum of the HX2 that trichloromethane kills immediately, is paved into uniform thin layer, cultivates 24 hours for 28 ℃, observes the appearance of inhibition zone.
Flat board to plant pathogenic fungi suppresses detection method.HX2 activates 48 hours down for 28 ℃ with improvement 523 inclined-planes, makes bacteria suspension with aseptic 0.85% physiological saline, determines that with Mike's turbidimetry whole bacteria suspension concentration is about 10
9CFU/mL.The fungi dibbling on the PDA flat board 25 ℃ the activation 72 hours, make it grow up to macrocolony, with the punch tool of 0.4cm beat getting carry disease germs the cultivation post (bacterium cake), put a slice bacterium cake in the centre of each PDA flat board.Accomplish fluently and the double-deck filter paper of sterilizing is placed on the PDA flat board using with the aperture punch tool, every ware is put four, and the every dull and stereotyped center of distance is 3cm, and two adjacent filter paper are 90 °.Adding 20 μ L HX2 bacteria suspensions on filter paper, is contrast to add physiological saline, and each handles 4 repetitions, trains sample down for 25 ℃ and measures the colony diameter of fungi after 4 days, 5 days, 6 days, 7 days respectively, and calculate and suppress growth rate.Suppressing growth rate calculates according to following formula:
Inhibition growth rate=(C-T)/C * 100%
C: contrast fungal growth diameter
T: fungal growth diameter behind the inoculation HX2
HX2 is as shown in table 1 to the dull and stereotyped inhibition of edaphic bacillus result.
Table 1 HX2 is to the dull and stereotyped (unit: mm) that suppresses of edaphic bacillus
| Strains A .tumefaciens | Antibacterial circle diameter Inhibition zone (dia.) | Strains A .rhizogenes | Antibacterial circle diameter Inhibition zone (dia.) | Strains A .vitis | Antibacterial circle diameter Inhibition zone (dia.) |
| C58 | 28.8±3.5 a | K27 | 26.4±1.9 | K308 | 33.8±3.3 |
| AC11 | 29.7±3.6 | K75 | 39.1±3.4 | G11(3) | 39.1±3.6 |
| B6S3 | 35.9±2.1 | NL12-2 | 41.3±2.5 | G7(2) | 25.8±3.1 |
| MG14-2 | 27.7±2.1 | B1 | 40.8±2.7 | MI23-1 | 31.5±2.1 |
| MG10-1 | 42.9±5.2 | D2 | 30.8±3.7 | MB26-1 | 27.3±1.5 |
| CY4 | 30.0±3.8 | ||||
| CY31 | 24.3±1.9 | ||||
| CY32 | 28.5±2.6 | ||||
| Pt12 | 30.5±3.7 | ||||
| Y5-1 | 39.8±2.1 | ||||
| MI4-6 | 38.0±2.8 |
Annotate: a represents standard deviation
HX2 is as shown in table 2 to the dull and stereotyped inhibition of other plant pathogenetic bacteria result.
Table 2 HX2 is to the dull and stereotyped (unit: mm) that suppresses of plant pathogenetic bacteria
| Plant pathogenetic bacteria | Antibacterial circle diameter (mm) | Plant pathogenetic bacteria | Antibacterial circle diameter (mm) |
| The former bacterium of cabbage black rot | 18.3±0 a | The former bacterium of bacterial blight of rice | 27.6±1.7 |
| The former bacterium of angular leaf spot of cucumber | 22.5±1.0 | The former bacterium of angular leaf spot of cotton | 28.9±1.9 |
| The former bacterium of canker of tomato | 39.0±2.1 |
Annotate: a represents standard deviation
HX2 is as shown in table 3 to the dull and stereotyped inhibition of plant pathogenic fungi result.
Table 3 HX2 suppresses plant pathogenic fungi is dull and stereotyped
| Fate | Plant the disease fungi | The bacterium colony contrast | Diameter aHandle | Suppress growth rate % | Plant the disease fungi | The bacterium colony contrast | Diameter is handled | Suppress growth rate % |
| 4 | Tomato is epidemic disease early | 45.2 | 35.0 | 22.8 | Cucumber is withered | 51.7 | 41.9 | 19.1 |
| Tomato gray mould | 51.3 | 40.7 | 20.8 | Watermelon is withered | 52.7 | 41.4 | 21.6 | |
| 5 | Tomato is epidemic disease early | 57.7 | 39.5 | 31.8 | Cucumber is withered | 65.2 | 47.3 | 27.6 |
| Tomato gray mould | 62.2 | 48.6 | 22.0 | Watermelon is withered | 66.0 | 49.0 | 25.9 | |
| 6 | Tomato is epidemic disease early | 70.8 | 53.1 | 39.3 | Cucumber is withered | 80.7 | 57.0 | 29.5 |
| Tomato gray mould | 77.7 | 60.3 | 21.9 | Watermelon is withered | 83.3 | 61.8 | 29.9 | |
| 7 | Tomato is epidemic disease early | 77.2 | 46.5 | 40.0 | Cucumber is withered | 82.3 | 60.5 | 26.6 |
| Tomato gray mould | 80.3 | 62.4 | 22.4 | Watermelon is withered | 84.0 | 67.3 | 20.0 |
Annotate: a represents diameter, unit: mm
Embodiment 3 greenhouses prevent and treat the root knot effect detection
HX2 is inoculated in the PDA liquid nutrient medium 28 ℃, and 150rpm shakes training 28 hours.Being mixed with concentration for the edaphic bacillus that tries is 10
8The bacteria suspension of CFU/mL mixes with the HX2 bacterium liquid and the stroke-physiological saline solution of 2 times of volumes respectively, compares with HX2 bacterium liquid and stroke-physiological saline solution inoculation simultaneously.On Sunflower Receptacle stem section, be vertical scuffing with the disinfection inoculation pin, make the wound of growing into 0.5cm, draw 3 μ L liquid to be measured and be inoculated in the wound, seal film parcel wound, cultivate in the greenhouse and observe the dross situation after 15 days and claim knurl heavy with parafilm.30 repetitions are established in every kind of processing, and this tests triplicate.
Preventive effect (%)=(it is on average heavy that contrast crown gall knurl on average weighs-handle the crown gall knurl) ÷ (contrast crown gall knurl is on average heavy) * 100%
The HX2 greenhouse prevents and treats root knot effectiveness results such as table 4
Table 4 HX2 greenhouse prevents and treats the root knot effect
| Strains A. tumefaciens | Preventive effect (%) | Strains A. rhizogenes | Preventive effect (%) | Strains A .vitis | Preventive effect (%) |
| C58 | 82.6 | K27 | 98.8 | K308 | 95.9 |
| AC11 | 80.6 | K75 | 98.8 | G11(3) | 81.6 |
| B6S3 | 39.8 | NL12-2 | 88.9 | G7(2) | 84.0 |
| MG14-2 | 96.1 | B1 | 96.1 | MI23-1 | 66.9 |
| MG10-1 | 66.7 | D2 | 100 | MB26-1 | 97.4 |
| CY4 | 100 | ||||
| CY31 | 100 | ||||
| CY32 | 80.1 | ||||
| Pt12 | 91.5 | ||||
| Y5-1 | 84.2 | ||||
| M14-6 | 100 |
Embodiment 4 greenhouses control Root of European Grape carninomatosis effect detection
HX2 is inoculated in the PDA liquid nutrient medium, and 28 ℃, 150mm shakes training 28 hours.Being mixed with concentration for grape edaphic bacillus (A.vitis) K308 that tries is 10
8The bacteria suspension of CFU/mL mixes with the HX2 bacterium liquid and the stroke-physiological saline solution of 2 times of volumes respectively.Each is handled and uses 4 strain grape stem sections, 4 inoculations of inoculation point between 3 leaf to the 7 leaves of every strain stem Duan Congdi, on the stem section, cause the wound of long 0.8cm with the disinfection inoculation pin, drawing 10 μ L liquid to be measured is inoculated in the wound, seal film parcel wound with parafilm, observe the dross situation after 30 days and claim knurl heavy.This test triplicate, preventive effect is calculated the same.HX2 reaches 99.5% to the preventive effect of the root knot that caused by grape edaphic bacillus (A.vitis) K308.
Embodiment 5 control in field Root of European Grape carninomatosis effect detection
District's group arrangement is at random adopted in the experimental plot, handles for every kind and repeats 4 times, and the sub-district completely random is arranged, 60 plant in every sub-district.Boundary belt is set to 3 meters.
Select for use the cuttage bar of the grape shearing of annual 2-3 joint to clean root with sterilized water; Pathogenic bacterium grape edaphic bacillus (A.vitis) K308 be inoculated in improvement 523 liquid nutrient mediums 28 ℃ of shaking tables cultivate 48 hours to the bacterium amount be 10
8CFU/mL, after biocontrol microorganisms HX2 microbial inoculum and germ K308 mix with 1: 1 (kilogram: rise), above-mentioned grape cuttage bar root being immersed bacterium liquid, take out plantation after 10 minutes, is contrast with the mixed solution of independent inoculation grape edaphic bacillus (A.vitis) K308 and turfy soil.Plant carries out conventional water and fertilizer management according to the practical situation on producing in the inoculation back.Grow after about 7 months (mid-April～mid-November), dig out, check that the formation of tumour and weighing knurl are heavy.Statistics sickness rate and preventive effect.
Plant sum * 100% of sickness rate (%)=(the plant quantity of the plant quantity+death of long knurl) ÷ plantation
Preventive effect (%)=(contrast sickness rate-processing sickness rate) ÷ contrasts sickness rate * 100%
The sickness rate of the Root of European Grape carninomatosis of inoculation Portugal's edaphic bacillus (A.vitis) K308 is 93.75% separately, and the disease plant tumour is bigger.The sickness rate of the Root of European Grape carninomatosis of inoculation HX2 microbial inoculum is 17.5%, and the tumour of disease plant is less, and preventive effect is 81.3%.Therefore, the HX2 microbial inoculum has good preventive effect to the Root of European Grape carninomatosis.
Embodiment 6 biological and ecological methods to prevent plant disease, pests, and erosion correlated character detect
Indolylacetic acid (IAA) detects.IAA is a plant hormone, can promote plant-growth, thereby improves disease resistance of plant.HX2 be inoculated in the NA substratum (peptone 5g/L, yeast extract 1.5g/L, extractum carnis 1.5g/L, NaCl 5g/L, pH7.0-7.2) and add in the 0.5g/L tryptophane NA substratum, 28 ℃ are shaken training sampling after 12 hours.Sampling 5mL, the centrifugal 10min of 12000rpm gets the 2mL supernatant, adds the 10mM phosphoric acid of 100 μ L, and 4mL detects liquid (the 0.5M FeCl of 1mL
3Be dissolved in 50mL of 35%HClO
4), use spectrophotometric instrumentation 530nm place's photoabsorption behind the room temperature reaction 25min.With pure product IAA obtain solution measurement standard absorption curve, and calculate the amount that HX2 produces IAA.HX2 cultivates the concentration that produces IAA after 12 hours in containing the NA substratum of tryptophane and reaches 24.8 μ g/mL.The concentration that produces IAA in not containing the NA substratum of tryptophane is 4.1 μ g/mL.Phosphate esterase active is measured.Phosphoesterase has the effect of branch phosphorus decomposing, can decompose the inorganic phosphorus in the soil after the biocontrol microorganisms that therefore produces phosphoesterase is used, thereby improves the content that plant in the soil can utilize phosphorus, plays the promotion plant-growth, improves the effect of disease resistance of plant.Use the dull and stereotyped phosphate esterase active that detects of Pikovskaya ' s.The dull and stereotyped compound method of Pikovskaya ' s is: yeast extract 0.5g/L, glucose 10g/L, calcium phosphate 5g/L, ammonium sulfate 0.5g/L, Repone K 0.2g/L, magnesium chloride 0.1g/L, manganous sulfate 0.0001g/L, ferrous sulfate 0.0001g/L, agar 15g/L is inoculated in the dull and stereotyped central authorities of Pikovskaya ' s to HX2, cultivates after 7 days for 28 ℃ and detects.Transparent circle occurs in periphery of bacterial colonies, show that HX2 has phosphate esterase active.
Embodiment 7 fermentation methods prepare HX2
1, actication of culture
Use improvement 523 substratum, culture medium prescription is: sucrose 10g/L, KH
2PO
42.0g/L, yeast extract paste 10g/L, CaCO
35.0gg/, NaCl 0.2g/L, MgSO
4.7H
2O 0.3g/L, agar 15g/L, pH.0-7.2.HX2 is inoculated on improvement 523 medium slant, cultivated 24 hours for 28 ℃.
2, culture of seed liquid
The composition of seed culture medium: sucrose 5g/L, corn steep liquor 2g/L, KH
2PO
40.05g/L, K
2HPO
40.05g/L, MgSO
47H
2O 0.025g/L, (NH
4)
2SO
40.5g/L, CaCO
32g/L, pH 7.0-7.2.The HX2 that activation is good is mixed with 10 with stroke-physiological saline solution
8The bacteria suspension of CFU/mL, the inoculum size with 1% is inoculated in the seed culture medium, and 28 ℃ of shaking table concussions are cultivated, and rotating speed is 150rpm, and incubation time is 16 hours.
3, ferment tank
Fermention medium consists of: glucose 10g/L, yeast extract paste 2g/L, NaCl 0.5g/L, KH
2PO
40.25g/L, K
2HPO
40.25g/L, MgSO
47H
2O 0.5g/L, (NH
4)
2SO
410g/L, pH7.0-7.2.Per minute air flow volume), tank pressure 0.05MPa cultured seed liquid is inserted in the fermentor tank with 2% inoculum size, and 28 ℃, stirring velocity is 180rpm, and air flow is 1: 0.6-0.8 (fermentating liquid volume:.Fermented 20 hours, the bacterium amount reaches 5-10 * 10
8CFU/mL obtains HX2 bacterium liquid.
Embodiment 8 microbial inoculums are made
Every liter and 4 kilograms turfy soils of HX2 bacterium liquid that embodiment 7 is fermented mix, and obtain the HX2 microbial inoculum.The product water content is 30-40%, and every gram microbial inoculum contains 1-2 * 10
8Aquatic engler bacterium (Rahnella aquatilis) HX2 that draws of CFU.
＜110〉China Agricultural University
＜120〉Rahnella aquatilis HX 2 and application thereof
<130>
<160>1
<170>PatentIn version 3.3
<210>1
<211>1491
<212>DNA
<213>Rahnella aquatilis HX2
<400>1
tgattacgaa ttcgagctcg gtacccgggg atcctctaga gattcaggcc taacacatgc 60
aagtcgagcg gcagcggaaa gtagcttgct actttgccgg cgagcggcgg acgggtgagt 120
aatgtctggg aaactgcctg atggaggggg ataactactg gaaacggtag ctaataccgc 180
atgacctcga aagagcaaag tgggggatct tcggacctca cgccatcgga tgtgcccaga 240
tgggattagc tagtaggtga ggtaatggct cacctaggcg acgatcccta gctggtctga 300
gaggatgacc agccacactg gaactgagac acggtccaga ctcctacggg aggcagcagt 360
ggggaatatt gcacaatggg cgcaagcctg atgcagccat gccgcgtgtg tgaagaaggc 420
cttagggttg taaagcactt tcagcgagga ggaaggcatc atacttaata cgtgtggtga 480
ttgacgttac tcgcagaaga agcaccggct aacttcgtgc cagcagccgc ggtaatacgg 540
agggtgcaag cgttaatcgg aattactggg cgtaaagcgc acgcaggcgg tttgttaagt 600
cagatgtgaa atccccgcgc ttaacgtggg aactgcattt gaaactggca agctagagtc 660
ttgtagaggg gggtagaatt ccaggtgtag cggtgaaatg cgtagagatc tggaggaata 720
ccggtggcga aggcggcccc ctggacaaag actgacgctc aggtgcgaaa gcgtggggag 780
caaacaggat tagataccct ggtagtccac gctgtaaacg atgtcgactt ggaggttgtg 840
cccttgaggc gtggcttccg gagctaacgc gttaagtcga ccgcctgggg agtacggccg 900
caaggttaaa actcaaatga attgacgggg gcccgcacaa gcggtggagc atgtggttta 960
attcgatgca acgcgaagaa ccttacctac tcttgacatc cacggaattc gccagagatg 1020
gcttagtgcc ttcgggaacc gtgagacagg tgctgcatgg ctgtcgtcag ctcgtgttgt 1080
gaaatgttgg gttaagtccc gcaacgagcg caacccttat cctttgttgc cagcacgtaa 1140
tggtgggaac tcaaaggaga ctgccggtga taaaccggag gaaggtgggg atgacgtcaa 1200
gtcatcatgg cccttacgag tagggctaca cacgtgctac aatggcatat acaaagagaa 1260
gcgaactcgc gagagcaagc ggacctcata aagtatgtcg tagtccggat tggagtctgc 1320
aactcgactc catgaagtcg gaatcgctag taatcgtaga tcagaatgct acggtgaata 1380
cgttcccggg ccttgtacat caccgcccgt cacaccatgg gagtgggttg caaaagaagt 1440
aggtagctta accttcggga gggcgcttac cactttgtga ttcatgactg g 1491
Claims (5)
1, aquatic engler bacterium (Rahnella aquatilis) the HX2 CGMCC No.1896 that draws.
2, the microbial inoculum that contains the described bacterial strain of claim 1.
3, the application of the described bacterial strain of claim 1 in biological control.
4, the application of the described bacterial strain of claim 1 in the control plant root cancer.
5, the application of the described bacterial strain of claim 1 in control Root of European Grape carninomatosis.
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| CNB2007100635207A CN100455659C (en) | 2007-02-02 | 2007-02-02 | Rahnella aquatilis HX2 and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100635207A CN100455659C (en) | 2007-02-02 | 2007-02-02 | Rahnella aquatilis HX2 and application thereof |
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| CN103266065A (en) * | 2013-05-15 | 2013-08-28 | 内蒙古农业大学 | Method for quickly culturing verticillium dahliae |
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| CN109234193B (en) * | 2018-09-26 | 2021-06-08 | 中国农业科学院蔬菜花卉研究所 | Rahnella aquatilis ZF7 and application thereof in plant growth promotion and disease prevention |
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