CN101928683B - Pseudomonas sp. strain and use thereof - Google Patents
Pseudomonas sp. strain and use thereof Download PDFInfo
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- CN101928683B CN101928683B CN2010100037262A CN201010003726A CN101928683B CN 101928683 B CN101928683 B CN 101928683B CN 2010100037262 A CN2010100037262 A CN 2010100037262A CN 201010003726 A CN201010003726 A CN 201010003726A CN 101928683 B CN101928683 B CN 101928683B
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- pseudomonas
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- bacterial wilt
- bacterium
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Images
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a pseudomonas sp. strain 2-29, which belongs to the class of pseudomonas sp. and was collected with a collection number of CGMCC No.3331 in China General Microbiological Culture Collection Center (abbreviated as CGMCC) of Institute of Microbiology Chinese Academy of Sciences located on Datun road in Chaoyang district of Beijing, on October 2009. The invention also provides the use of the pseudomonas sp. strain 2-29 in the prevention and control of tomato ralstonia solanacearum. For solving the problem that an effective chemical for treating tomato ralstonia solanacearum lacks at present and the problem that pseudomonas sp. is susceptible to generating medicament resistance to chemical pesticide, a strain having an antagonistic effect on tomato ralstonia solanacearumis screen through screening tomato ralstonia solanacearum antagonists and the test of the control effect of the tomato ralstonia solanacearum antagonists; on this basis, field tests can be further performed for developing a mature microbial fertilizer; and thus, the pseudomonas sp. strain 2-29 has a great significance for reducing the use of a large amount of fertilizer and pesticide and reducing agricultural pollution sources.
Description
Technical field
The present invention relates to biological field, relate in particular to a kind of pseudomonad strain and application thereof.
Background technology
Bacterial wilt of tomato is to cause worldwide crushing soil-borne disease by the blue or green withered La Er Salmonella of eggplant [Ralatonia solanacearum (smith) Yabuuchi et al]; Tropical, semi-tropical countries and regions; As: South America, Japan, India, Philippines, area such as China Taiwan, Guangdong, Zhejiang, Fujian, Jiangsu, Hubei, Sichuan, Chongqing is serious to take place.It is dead that this disease can cause the plant big area to wilt, and general field piece sickness rate is 10%～15%, and grave illness field sickness rate causes tomato yield seriously to descend up to 80%～98.5%, causes great financial loss.
R.solanacearum is a kind of soil inhabitant, can in the soil under no host's situation, survive for a long time.R.solanacearum mainly invades plant from root system, also can invade and directly get into conduit system from stem's wound, and a large amount of therein breedings also produce metabolic substd, hinder moisture transportation in the plant materials, cause the plant wilting.At present, lack the effective chemical medicament to bacterial wilt of tomato, Ralstonia solanacearum is prone to develop immunity to drugs to chemical pesticide in addition, and uses chemical pesticide can cause environmental pollution and production cost to rise in a large number.The phase of lying fallow spreads fertilizer over the fields lime and can alleviate the disease generation, but is prone to cause soil compaction.Breeding for disease resistance work owing to lack the ideal anti-source material, is made slow progress.Rice field-upland field rotation requires crop rotation year limit for length, and near other host of nothing exists and can play good result, thereby implements difficulty.Formed the soil organisms vacuum behind the soil disinfection,, can cause the great outburst of pathogenic bacteria and cause even more serious consequence in case soil receives pathogen infection.The biological control crop pest have available stock abundant, with low cost, can avoid secondary environmental pollution, be considered to solve approach likely of soil-borne disease.Utilize the pathogenic bacteria antagonistic microbe to carry out the important component part that biological control is biological control.
Summary of the invention
2.-29 technical problem to be solved by this invention has provided a kind of pseudomonad strain of utilizing pathogenic bacteria antagonistic microbe control of plant bacterial wilt; Its called after pseudomonas (Pseudomonas sp.) of classifying; Its deposit number is CGMCC NO.3331, and its preservation time is on October 9th, 2009, and depositary institution is China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC); The address is the Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
The present invention realizes through following technical scheme:
Separate a kind of pseudomonad strain 2.-29:
The screening step is following:
Gather 21 soil samples from tomato planting bases such as Jiashan, Zhejiang, Lishui, Hangzhou in (1) 2008 year, and utilized beef-protein medium to separate, be separated to bacterium strain more than 420 altogether.
(2) carry out dull and stereotyped antagonistic effect then, obtain the blue or green withered La Er Salmonella of bacterial wilt of tomato eggplant is had primary dcreening operation bacterium 27 strains of better antagonistic effect.
(3), further sieve to such an extent that the blue or green withered La Er Salmonella of bacterial wilt of tomato eggplant is had metastable antagonistic effect sieve bacterium 4 strains again through pot experiment.
2.-29 this bacterial strain sieves the strain bacterium in the bacterium for above-mentioned 4 strains again.According to morphological specificity, cultural characteristic and the physiological and biochemical property of bacterial strain,, 2.-29 this bacterial strain is identified that tentatively confirm as the genus Rhodopseudomonas, this bacterial strain sampling separates from Lishui of Zhejiang with reference to uncle Jie Shi Bacteria Identification handbook.
Said pseudomonad strain is 2.-29: belong to pseudomonas (Pseudomonas sp.); Well-grown in beef-protein medium, violet staining is inhomogeneous, the cell rod-short; Bacterium colony is circular when on the beef extract-peptone agar plate, cultivating, neat in edge and rule, and surface wettability, smooth, translucent, the khaki color of bacterium colony, lawn is thin.
The present invention also provides said pseudomonad strain 2.-29 in the application of control of plant bacterial wilt.The present invention is directed to present bacterial wilt of tomato and lack the effective chemical medicament; Ralstonia solanacearum problem that chemical pesticide is prone to develop immunity to drugs through to Antagonistic Strains Against Tomato Bacterial Wilt separation screening and efficiency test thereof, separates the bacterial strain that bacterial wilt of tomato is had antagonistic action in addition; Can carry out further field test on this basis; So as to developing a kind of sophisticated microbial organic fertilizer,, alleviate widespread pollution from the overuse of fertilizers and pesticides in rural area and have important effect reducing the use of a large amount of chemical fertilizer, agricultural chemicals.
Description of drawings
Fig. 1 is 2.-29 antagonistic effect synoptic diagram of the withered La Er Salmonella of pair green grass or young crops of bacterial strain among the present invention.
Fig. 2 is that tomato seedling is transplanted engagement CK1 and the potted plant effect synoptic diagram of CK2 after the 15th day among the present invention.
Fig. 3 be among the present invention tomato seedling transplant engagement after the 15th day bacterial strain 2.-29 handles and contrasts the potted plant effect synoptic diagram of CK2.
Fig. 4 is an Antagonistic Strains Against Tomato Bacterial Wilt biocontrol effect test chart among the present invention.
Embodiment
Through following examples the present invention is described in more detail, but following examples only are illustrative that the present invention does not receive the restriction of these embodiment.
Explanation to related material:
Peptone: biochemical preparation, Sangon Biotech (Shanghai) Co., Ltd.;
Carnis Bovis seu Bubali cream: biochemical preparation, the extensive and profound in meaning star biotechnology in Beijing Ltd;
Yeast extract (yeast extract): biochemical reagents, Shanghai yeast factory;
Casein hydrolysate (Lip river proteolysate): biochemical reagents, BIO.BASIC.INC.;
Agar: biochemical reagents, Quanzhou City, Fujian spring Hong Kongnization factory;
NaCl: analytical pure, Shanghai green grass or young crops is analysed Chemical Industry Science Co., Ltd;
K
2HPO
4: analytical pure, BIO.BASIC.INC.;
Glucose: analytical pure, Sangon Biotech (Shanghai) Co., Ltd.;
MgSO
47H
2O: analytical pure, Chinese ilex chemical plant;
Sucrose: analytical pure, BIO.BASIC.INC..
Embodiment 1: the separation and purification of bacterium
By following testing sequence bacterium is carried out separation and purification:
1 substratum
Beef extract-peptone nutrient agar: Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g, agar 15-20g, H
2O 1000ml, PH 7.0-7.2.121.3 ℃ sterilization 20 minutes.
2 testing sequences
2.1 the preparation of substratum
Preparation as 1 described beef extract-peptone nutrient agar, preparation bacterium slant medium.
2.2 the preparation of soil diluent
(1). take by weighing 10g soil, add 100ml and have in the sterilized water 500ml triangular flask of granulated glass sphere, vibrated 15 minutes, i.e. 10-1;
(2). draw soil suspension 10ml, add in the 90ml sterilized water 500ml triangular flask, promptly 10
-2
(3). from (2), draw 10ml, add in the 90ml sterilized water 500ml triangular flask, promptly 10
-3
(4). from (3), draw 5ml, add in the 45ml sterilized water 250ml triangular flask, promptly 10
-4
(5). from (4), draw 5ml, add in the 45ml sterilized water 250ml triangular flask, promptly 10
-5
(6). from (5), draw 5ml, add in the 45ml sterilized water 250ml triangular flask, promptly 10
-6
2.3 bacterium separates
(7). from (3) (4) (5), draw 0.1ml respectively, join and be coated with cultivation (cultivating 2-3 days for 28-30 ℃) in the beef extract-peptone nutrient agar flat board, 3 repetitions respectively are set;
(8). the petridish of selection 20-200 colony count carries out picking colony and carries out slant culture (cultivating 2-3 days for 28-30 ℃);
2.4 many soil samples
(9). totally 21 soil samples, each soil sample repeats above-mentioned 2.1～2.3 steps.
2.5 result
Separate as stated above and obtain bacterium strain more than 420.
Embodiment 2: the bacterium that separation obtains is to the dull and stereotyped antagonistic effect of the blue or green withered La Er Salmonella of bacterial wilt of tomato eggplant
1. substratum
1.1 solid medium
Ralstonia solanacearum substratum preparation: MgSO
47H
2O0.3g, K
2HPO
42.0g, Yeast extract (yeast extract) 4.0g, Casein hydrolysate (Lip river proteolysate) 8.0g, Sucrose (sucrose) 10.0g, Agar (agar) 18g, Distilled water (zero(ppm) water) 1000ml.
Beef extract-peptone nutrient agar preparation: with among the embodiment 11.
1.2 liquid nutrient medium
Bacterial liquid shake-flask culture base: beef-protein medium does not add agar and gets final product.
Bacterial wilt of tomato bacteria liquid shake-flask culture base: the ralstonia solanacearum substratum does not add agar and gets final product.
2, testing sequence
2.1 medium preparation
Preparation ralstonia solanacearum substratum (solid, liquid), the blue or green withered La Er Salmonella of preparation pathogenic bacteria eggplant inclined-plane; Preparation beef extract-peptone nutrient agar (solid, liquid), preparation bacterium inclined-plane.
2.2 dull and stereotyped antagonistic effect
2.2.1 actication of culture
Bacterial wilt of tomato bacterium and bacterium carry out actication of culture: bacterial wilt of tomato bacterium and separation of bacterial are switched to ralstonia solanacearum medium slant and beef extract-peptone nutrient agar inclined-plane, and 28-30 ℃, cultivated 16-28 hour, subsequent use.
2.2.2 the preparation ralstonia solanacearum is dull and stereotyped
The activation ralstonia solanacearum inserts in the liquid nutrient medium and carries out shake-flask culture, and 28-30 ℃, 200 rev/mins.Dull and stereotyped with the ralstonia solanacearum substratum every other day, (concentration is 10 etc. adding 0.1ml in each flat board after the culture medium solidifying
9～10
10Individual/ml) ralstonia solanacearum fermented liquid (fermenting about 40 hours), coating, 28-30 ℃ of overnight cultures, subsequent use.
2.2.3 prepare the bacterial liquid fermented liquid
Cultivated second day at the ralstonia solanacearum liquid shaking bottle, the activation separation of bacterial inserts in the liquid nutrient medium and carries out shake-flask culture, and 28-30 ℃, 200 rev/mins, subsequent use every other day (fermenting about 40 hours).
2.2.4 dull and stereotyped antagonistic effect
Dip in sterilization filter paper (Φ 9mm) and to get the separation of bacterial fermented liquid and put into the ralstonia solanacearum flat board, each flat board is put the different antagonistic bacteriums of 5 strains, 3 repetitions.24h left and right sides observation experiment result mainly investigates the inhibition zone size that has that it's too late.
3, result
Obtain antagonistic effect bacterium 27 strains preferably through dull and stereotyped antagonistic effect, 2.-29 bacterial strain is exactly a wherein strain.Referring to Fig. 1.Fig. 1 experiment is for being coated with earlier the blue or green withered La Er Salmonella fermented liquid of 0.1ml on flat board, then with filter paper dip in get bacterial strain 2.-29 fermented liquid be placed on the flat board, cultivate simultaneously and get.
Can find out that by Fig. 1 inhibition zone is obvious, 2.-29 fast growth of bacterial strain is described, diffusibility is strong, to the blue or green withered La Er Salmonella tool of eggplant antagonistic action significantly.
Embodiment 3: bacterial strain 2.-29 fermented liquid carries out the biocontrol effect test in order to check bacterial strain biocontrol effect 2.-29 to bacterial wilt of tomato on potted plant, carry out this test.
Experimental procedure:
1, grows seedlings
1.1 the preparation of seedling medium
Seedling medium is: the peat composed of rotten mosses: vermiculite: perlite=7: 1: 0.5, regulate water ratio to 60%～70% with tap water then, and subsequent use.
1.2 sowing
1.1 about 3/4ths height of in the every cave of seedling culture hole plate (specification 90 * 60), packing into preparations matrix is pressed with have gentle hands, evenly is sprinkled into 50 tomato seeds in every then cave, the very thin matrix of layer overlay again, and cover the cave dish with plastics bag, and prevent moisture evaporation, subsequent use.
1.3 emerge
Put into growth cabinet to 1.2 ready sowing caves dish and cultivate, 25 ℃ of temperature are put into Da Penshui and are preserved moisture, and open all fluorescent lamps, all turn off later on for 18, and guarantee that day and night the temperature difference is more than 10 ℃.Emerged in about 4～5 days, and took plastics bag off after emerging, wait two cotyledons open and flat, subsequent use.
2, seedling moves in the dish of cave and cultivates
2.1 transplantation of seedlings
In dish (specification 45 * 45) every cave, cave, fill by 1.1 preparation matrix (slightly compressing), move into the open and flat seedling of 1 strain, 1.3 ready two cotyledons in every then cave, two refer to slightly compress at root, in booth, cultivate.Be cultured to 3～4 true leaves (about 3～4 weeks) in the dish of cave.Attention: two refer to pinch 1 cotyledon when taking seedling.
2.2 blue or green withered La Er Salmonella is prepared with test antagonism fermented liquid
Blue or green withered La Er Salmonella and the activation of test antagonism bacterium, every then strain connects liquid shaking bottle and cultivates, and 28～30 ℃, 200 rev/mins, cultivate about 40 hours, subsequent use.
2.3 use the antagonism bacterium in advance
Preceding 1 week of transplantation of seedlings engagement, add the corresponding antagonism fermented liquid of 5ml in every cave.
3, potted plant
3.1 potting soil is prepared
2～4 tons of soil are dug in the serious plot of bacterial wilt morbidity, Nian Qiancong Lishui of Zhejiang tomato planting base, and are subsequent use.
3.2 soil dress basin
Take by weighing 2.1 of appropriate amount and prepare, sneak into 2% fertilizer, pack in every basin (specification 280 * 220) 4.5 kilograms and mix soil, subsequent use.
3.3 transplant
Transplant 5 strain tomato seedlings in every basin, keep the skin wet as required.2 contrasts are established in this test, i.e. CK1 and CK2, CK1: do not add any bacterium during transplanting; CK2: the blue or green withered La Er Salmonella dilution fermented liquid (diluting 500 times) of 50ml is only watered in every strain during transplanting.2.-29 bacterial strain is handled, and each handles 6 basins, every basin 5 young plants, and uniform distribution, every strain root waters the blue or green withered La Er Salmonella dilution fermented liquid (diluting 500 times) of 50ml earlier during transplanting, and then waters 50ml antagonism bacterium dilution fermented liquid (diluting 10 times).
3.4 record
Write down temperature every day, humidity, standard plant true leaf number, every basin morbidity strain number, incidence of leaf and total true leaf number.
4, result
(annotate: transplanted engagement on May 17th, 2009, photo is to take on May 31st, 2009) as shown in Figures 2 and 3.Fig. 2 and Fig. 3 photo are that tomato seedling is transplanted the photo that engagement was taken on the 15th day, and on May 31st, 1 took.Can find out that by Fig. 4 CK1 is an acomia diseased plant on June 22nd, 2009 to observing end; CK2 fell ill since on May 21st, 2009, reached 24 strains to morbidity strain on May 31st, 2009 number; 2.-29 bacterial strain is handled since on May 19th, 2009 and is fallen ill; Also have only 9 strains to morbidity strain on May 31st, 2009 number; 2.-29 pair certain preventive effect of bacterial wilt of tomato tool of bacterial strain is described; Though 2.-29 bacterial strain is handled the diseased plant number still to compare photograph CK2 few when observe finishing on June 22nd, 2009, not obvious to later stage preventive effect effect.Reason possibly be that blue or green at leisure withered La Er Salmonella breeds in a large number, and bacterial strain 2.-29 can't prevention and control.Therefore when suggestion used this bacterial strain 2.-29 to carry out the bacterial wilt control, the later stage mended with 2.-29 goods of bacterial strain; Perhaps using goods of the present invention combines the effect meeting better with the measure of plantation gardening.
Embodiment 4: the bacterium to separation obtains is identified
According to bacterial strain morphological specificity, cultural characteristic and physiological and biochemical property 2.-29,, 2.-29 this bacterial strain is identified that tentatively confirm as the genus Rhodopseudomonas, this bacterial strain sampling separates from Lishui of Zhejiang with reference to uncle Jie Shi Bacteria Identification handbook.
Characteristics such as the form of this preservation strain, cultivation, physiology, biochemistry are following:
Pseudomonad strain is 2.-29: belong to pseudomonas (Pseudomonas sp.); Well-grown in beef-protein medium, violet staining is inhomogeneous, the cell rod-short; Bacterium colony is circular when on the beef extract-peptone agar plate, cultivating, neat in edge and rule, and surface wettability, smooth, translucent, the khaki color of bacterium colony, lawn is thin.
Bacterial strain is physics and chemistry test-results (BIOLOG GN2) 2.-29
| Test subject | The result | Test subject | The result | Test subject | The result | Test subject | The result |
| Cellular form | Shaft-like | Gramstaining | Negative | Oxydase | - | Catalase | + |
| Contrast | - | The D-melibiose | - | The p-phenylglycollic acid | + | The L-Histidine | + |
| The a-Schardinger dextrins | - | Methyl glucoside | - | Methylene-succinic acid | - | Ls-hydroxyproline | + |
| Dextrin | - | The D-psicose | - | The a-ketobutyric acid | - | The L-leucine | + |
| Glycogen | - | The D-raffinose | - | The a-keto-glutaric acid | + | The L-ornithine | + |
| Tween40 | - | The L-rhamnosyl | - | The a-oxopentanoic acid | - | The L-phenylalanine(Phe) | + |
| Tween80 | B | The D-sorbyl alcohol | - | D, L-lactic acid | + | The L-proline(Pro) | + |
| N-acetylgalactosamine | - | Sucrose | - | Propanedioic acid | - | The L-Pyrrolidonecarboxylic acid | + |
| N-acetyl grape osamine | - | The D-trehalose | - | Propionic acid | + | The D-Serine | + |
| Ribitol | - | Turanose | - | Quinic acid | + | The L-Serine | + |
| L-arabinose | - | Xylitol | - | The D-saccharinic acid | + | The L-Threonine | + |
| The D-arabitol | - | Pyruvic Acid Methyl ester | + | The certain herbaceous plants with big flowers diacid | - | D, the L-carnitine | + |
| The D-cellobiose | - | The Succinic Acid methyl esters | + | Succinic Acid | + | The g-propalanine | + |
| The i-tetrahydroxybutane | - | Acetate | + | Bromosuccinic acid | + | Urocanic acid | + |
| D-fructose | B | Suitable-aconic acid | + | Succinamic acid | + | Inosine | + |
| L-fructose | - | Hydrocerol A | + | Glucuronamide | - | Uridine | - |
| The D-semi-lactosi | - | Formic acid | - | The L alanimamides | + | Thymidine | - |
| Gentiobiose | - | The D-galactonolactone | - | The D-L-Ala | + | Phenyl-ethyl amine | + |
| A-D-glucose | + | The D-galacturonic acid | + | The L-L-Ala | + | Putrescine | + |
| The m-inositol | - | The D-saccharic acid of crawling | + | The L-alanyl-glycine | + | The 2-monoethanolamine | + |
| The a-D-lactose | - | The b-methyl glucoside | - | Altheine | + | 2, the 3-butyleneglycol | B |
| Lactulose | - | The D-uronic acid of crawling | + | The L-aspartic acid | + | Glycerine | + |
| SANMALT-S | - | The a-hydroxybutyric acid | - | L-L-glutamic acid | + | D, the L-glycerophosphate | - |
| D-N.F,USP MANNITOL | - | The b-hydroxybutyric acid | + | The glycyl aspartic acid | - | D-sugar-1-the phosphoric acid of crawling | - |
| The D-seminose | + | The g-hydroxybutyric acid | - | Glycyl L-glutamic acid | + | The D-Robison ester | B |
Bacterial strain 2.-2916S rRNA gene sequencing result (SEQ ID No.1 in the sequence table) as follows:
GGTAACCGTCCCCCCGAAGGTTAGACTAGCTACTTCTGGTGCAACCCACTCCC
ATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCAACATT
CTGATTTGCGATTACTAGCGATTCCGACTTCACGCAGTCGAGTTGCAGACTGC
GATCCGGACTACGATCGGTTTTGTGAGATTAGCTCCACCTCGCGGCTTGGCAA
CCCTCTGTACCGACCATTGTAGCACGTGTGTAGCCCAGGCCGTAAGGGCCATG
ATGACTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCCTTAG
AGTGCCCACCATAACGTGCTGGTAACTAAGGACAAGGGTTGCGCTCGTTACGG
GACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCAGCACCTG
TGTCAGAGTTCCCGAAGGCACCAATCCATCTCTGGAAAGTTCTCTGCATGTCA
AGGCCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGC
TTGTGCGGGCCCCCGTCAATTCATTTGAGTTTTAACCTTGCGGCCGTACTCCCC
AGGCGGTCAACTTAATGCGTTAGCTGCGCCACTAAAATCTCAAGGATTCCAAC
GGCTAGTTGACATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGC
TCCCCACGCTTTCGCACCTCAGTGTCAGTATCAGTCCAGGTGGTCGCCTTCGC
CACTGGTGTTCCTTCCTATATCTACGCATTTCACCGCTACACAGGAAATTCCAC
CACCCTCTACCATACTCTAGCTCGCCAGTTTTGGATGCAGTTCCCAGGTTGAGC
CCGGGGCTTTCACATCCAACTTAACGAACCACCTACGCGCGCTTTACGCCCAG
TAATTCCGATTAACGCTTGCACCCTCTGTATTACCGCGGCTGCTGGCACAGAGT
TAGCCGGTGCTTATTCTGTCGGTAACGTCAAAACAGCAAGGTATTAACTTACTG
CCCTTCCTCCCAACTTAAAGTGCTTTACAATCCGAAGACCTTCTTCACACACG
CGGCATGGCTGGATCAGGCTTTCGCCCATTGTCCAATATTCCCCACTGCTGCCT
CCCGTAGGAGTCTGGACCGTGTCTCAGTTCCAGTGTGACTGATCATCCTCTCA
GACCAGTTACGGATCGTCGCCTTGGTGAGCCATTACCTCACCAACTAGCTAATC
CGACCTAGGCTCATCTGATAGCGCAAGGCCCGAAGGTCCCCTGCTTTCTCCCG
TAGGACGTATGCGGTATTAGCGTTCCTTTCGAAACGTTGTCCCCCACTACCAGG
CAGATTCCTAGGCATTACTCACCCGTCCGCCGCTGAATCGAAGAGCAAGCTCT
CCTCATCCGCTCGACTTGC
16S rRNA gene order is carried out the Blast interpretation of result; Obtain reaching 99% with pseudomonas type strain similarity; Therefore according to 16S rRNA gene sequencing result, this bacterial strain can be classified as pseudomonas putida, but physiological and biochemical index and pseudomonas putida differ greatly; At present also do not have suitable type strain corresponding with it, belong to new bacterial strain.
Sequence table
< 110>Zhejiang University
The Zhejiang Academy of Agricultural Science
< 120>a kind of pseudomonad strain and application thereof
<130>1
<160>1
<170>PatentIn?version?3.3
<210>1
<211>1406
<212>DNA
<213>Pseudomonas?sp.
<400>1
ggtaaccgtc?cccccgaagg?ttagactagc?tacttctggt?gcaacccact?cccatggtgt 60
gacgggcggt?gtgtacaagg?cccgggaacg?tattcaccgc?aacattctga?tttgcgatta 120
ctagcgattc?cgacttcacg?cagtcgagtt?gcagactgcg?atccggacta?cgatcggttt 180
tgtgagatta?gctccacctc?gcggcttggc?aaccctctgt?accgaccatt?gtagcacgtg 240
tgtagcccag?gccgtaaggg?ccatgatgac?ttgacgtcat?ccccaccttc?ctccggtttg 300
tcaccggcag?tctccttaga?gtgcccacca?taacgtgctg?gtaactaagg?acaagggttg 360
cgctcgttac?gggacttaac?ccaacatctc?acgacacgag?ctgacgacag?ccatgcagca 420
cctgtgtcag?agttcccgaa?ggcaccaatc?catctctgga?aagttctctg?catgtcaagg 480
cctggtaagg?ttcttcgcgt?tgcttcgaat?taaaccacat?gctccaccgc?ttgtgcgggc 540
ccccgtcaat?tcatttgagt?tttaaccttg?cggccgtact?ccccaggcgg?tcaacttaat 600
gcgttagctg?cgccactaaa?atctcaagga?ttccaacggc?tagttgacat?cgtttacggc 660
gtggactacc?agggtatcta?atcctgtttg?ctccccacgc?tttcgcacct?cagtgtcagt 720
atcagtccag?gtggtcgcct?tcgccactgg?tgttccttcc?tatatctacg?catttcaccg 780
ctacacagga?aattccacca?ccctctacca?tactctagct?cgccagtttt?ggatgcagtt 840
cccaggttga?gcccggggct?ttcacatcca?acttaacgaa?ccacctacgc?gcgctttacg 900
cccagtaatt?ccgattaacg?cttgcaccct?ctgtattacc?gcggctgctg?gcacagagtt 960
agccggtgct?tattctgtcg?gtaacgtcaa?aacagcaagg?tattaactta?ctgcccttcc 1020
tcccaactta?aagtgcttta?caatccgaag?accttcttca?cacacgcggc?atggctggat 1080
caggctttcg?cccattgtcc?aatattcccc?actgctgcct?cccgtaggag?tctggaccgt 1140
gtctcagttc?cagtgtgact?gatcatcctc?tcagaccagt?tacggatcgt?cgccttggtg 1200
agccattacc?tcaccaacta?gctaatccga?cctaggctca?tctgatagcg?caaggcccga 1260
aggtcccctg?ctttctcccg?taggacgtat?gcggtattag?cgttcctttc?gaaacgttgt 1320
cccccactac?caggcagatt?cctaggcatt?actcacccgt?ccgccgctga?atcgaagagc 1380
aagctctcct?catccgctcg?acttgc 1406
Claims (6)
1. 2.-29 a pseudomonad strain is characterized in that: its called after pseudomonas (Pseudomonas sp.) of classifying, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and its deposit number is CGMCC No.3331.
2. 2.-29 a kind of pseudomonad strain as claimed in claim 1 is characterized in that said pseudomonad strain 16S rRNA gene order 2.-29 is SEQ ID NO:1.
3. a kind of pseudomonad strain as claimed in claim 1 2.-29; 2.-29 characteristic is following to it is characterized in that said pseudomonad strain: the strain cell rod-short; Bacterium colony is circular when on the beef extract-peptone agar plate, cultivating, neat in edge and rule, surface wettability, smooth; Translucent, the khaki color of bacterium colony, lawn is thin.
4. according to claim 1 or claim 2 2.-29 application in control of plant bacterial wilt of a kind of pseudomonad strain.
5. 2.-29 application in control of plant bacterial wilt of a kind of pseudomonad strain as claimed in claim 4 is characterized in that its being applied as in said control of plant bacterial wilt: with concentration is 10
9～10
10The pseudomonas of individual/ml 2.-29 liquid fermentation liquid is seeded to that to contain 0.1ml concentration be 10
9～10
10In the plate culture medium of the bacterial wilt of tomato fermented liquid of individual/ml, cultivate 24h, bacterial wilt of tomato is had antagonistic action.
6. 2.-29 application in control of plant bacterial wilt of a kind of pseudomonad strain as claimed in claim 4 is characterized in that its being applied as in said control of plant bacterial wilt: with inoculum size is that every gram soil contains 10 approximately
6～10
7Individual antimicrobial pseudomonas short of money 2.-29 liquid fermentation liquid is seeded in the basin of planting tomato seedling, potted plant in every gram soil contain 10 approximately
5～10
6Individual bacterial wilt of tomato bacterium was cultivated 15 days, and inoculation has 2.-29 tomato seedling morbidity strain number minimizing of bacterium liquid of pseudomonas.
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| CN2010100037262A CN101928683B (en) | 2010-01-01 | 2010-01-01 | Pseudomonas sp. strain and use thereof |
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| CN106047741B (en) * | 2016-03-04 | 2019-09-27 | 李涛 | It is a kind of to utilize the bacterial strain and synthetic method of chitin synthesis PHA and its application |
| CN106490003B (en) * | 2016-08-31 | 2019-02-01 | 浙江省农业科学院 | Inhibition and its extracting method of the celery root exudates concentrate to ralstonia solanacearum of tomato |
| CN106978373B (en) * | 2017-05-09 | 2019-07-09 | 福建农林大学 | One plant of pseudomonad and application for bacteriosis prevention and treatment |
| CN109603515A (en) * | 2019-01-16 | 2019-04-12 | 郑州大学 | A kind of microbial deodorant and preparation method thereof for benzene homologues |
| CN112522136A (en) * | 2020-11-19 | 2021-03-19 | 贵州省烟草科学研究院 | Pseudomonas and application thereof in controlling bacterial wilt of solanaceae crops |
| CN114561324B (en) * | 2022-03-03 | 2023-04-07 | 山西农业大学 | Tomato bacterial wilt antagonistic strain and application thereof in prevention and treatment of tomato bacterial wilt |
| CN114657101B (en) * | 2022-04-06 | 2023-06-09 | 华南农业大学 | Pseudomonas aeruginosa Q4-3 strain of tray Huo Ni and application thereof in treating and/or preventing bacterial wilt |
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| CN1515666A (en) * | 2000-10-11 | 2004-07-28 | 常州兰陵制药有限公司 | Method for screening pseudomonads for controlling blight |
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| CN1515666A (en) * | 2000-10-11 | 2004-07-28 | 常州兰陵制药有限公司 | Method for screening pseudomonads for controlling blight |
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