CN108441456A - One pseudomonas and its separation method and application - Google Patents
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Abstract
The present invention relates to a pseudomonas and its separation method and applications.For the pseudomonas strains on April 10th, 2018 in China typical culture collection center preservation, preservation is entitled：Entomophila pseudomonad Pseudomonas entomophila 1257, deposit number are CCTCC No：M 2018189.The pseudomonad of the present invention has significant inhibition to xanthomonas oryzae pv. oryzicola, while also having inhibition to rice leaf spot bacteria and Pyricularia oryzae.
Description
Technical field
The invention belongs to microorganisms technical fields, more particularly, to a pseudomonas and its separation method and application.
Background technology
Xanthomonas oryzae pv. oryzicola (Xanthomonas oryzae pv.oryzicola, Xoc) is the cause under rice Xanthomonas campestris kind
Lesion kind causes bacterial stripe (Bacterial leaf streak, BLS), is bacteriosis important on rice.It should
Disease mostly occurs, and in the Rice Croppings region such as Asia and Africa, (Zhang Rongsheng, Liu Yongfeng, Chen Zhi friendship xanthomonas oryzae pv. oryzicolas are short of money
Antibacterial screening, evaluation and application study [J] Chinese biological preventing and treating journals, 2011,27 (04):510-514.；
Rongsheng Zhang,Zhiyi Chen,Yongfeng Liu.Advance in Rice Bacterial Leaf Streak
Researches[J].Agricultural Science&Technology,2015,16(02):298-305+310.), especially
It is in subtropical and tropical zones (Adhikari T.B., and Mew T.W., Pathogenic variability in
Xanthomonas compestriis pv.oryzicola, IRRN., 1985,10 (1), 16), such as Malaysia, Vietnam, print
Degree etc..China be mainly distributed on Zhejiang on the south the Changjiang river, Fujian, Guangdong, Hainan, Guangxi, Yunnan, Jiangxi, Hunan, Guizhou,
The areas such as Sichuan, Hubei.In recent years, which is on the rise, and has evolved into the fourth-largest disease on rice.Population of Xanthomonas Oryzae Pv with
The interaction of host rice does not meet the relationship of gene-for-gene.In production, it has not been found that having immune water to Population of Xanthomonas Oryzae Pv
The rice varieties of rice varieties, general water resistant rice bacterial leaf spot pathogenic bacteria do not show resistance to Population of Xanthomonas Oryzae Pv.Therefore, searching being capable of antagonism item
The beneficial microbe of pinta bacterium is used for biological control, controls the generation of disease, be an ideal approach.
Biological control is mainly by using to non-phytotoxic or beneficial microorganism and its metabolic active substance, to influence
Or inhibit the existence and activity of phytopathogen, to effectively inhibit the generation of disease.The technology have do not allow to be also easy to produce resistance,
It free from environmental pollution, the advantages that people and other biological safety and biological and ecological methods to prevent plant disease, pests, and erosion material are easy to obtain, is sent out in integrated pest control
Wave important role.As subtracting for chemical pesticide apply, subtracts use, biological control will gradually replace traditional chemical prevention means,
With more wide application prospect (Li Xinglong, Li Yan loyalty soil-borne diseases Research Progress on Biological Control [J] Acta Pratacultures,
2015,24(03):204-212.)。
Pseudomonad (Pseudomonas spp.) is the quasi-microorganism being active in plant rhizosphere, and some of them are energy
Enough promote plant growth and the biocontrol microorganisms with controlling plant diseases, referred to as plant rhizosphere growth-promoting bacterium (Sun Guangzheng, Yao Tuo, Zhao
Gui Qin, Lu Hu, refined Pseudomonas fluorescens controlling plant diseases status and prospectives [J] the Acta Pratacultures of Marvin's, 2015,24
(04):174-190.)。
Entomophila pseudomonad is a kind of biocontrol microorganisms (Hu Jiangchun, Xue Delin, Ma Chengxin, king's book brocade plant roots for capableing of desinsection
The research of border Promoting bacteria (Pgpr) and application prospect [J] Chinese Journal of Applied Ecologies, 2004,15 (10):4.) it, was metabolized in growth
Cheng Zhong, will produce and secrete a variety of antibacterial substances, these active materials, which may also have, inhibits pathogenetic bacteria and fungi
Effect, Biocontrol Potential is up for further developing.
Invention content
The purpose of the present invention is just to provide for a pseudomonas and its separation method and application.The false unit cell of the present invention
Bacterium has significant antagonism to Xanthomonas oryzae pv. oryzicola.
The purpose of the present invention can be achieved through the following technical solutions：
The first aspect of the present invention is：One plant of entomophila pseudomonad is provided, Pseudomonas entomophila are named as
1257 or entomophila pseudomonad 1257, the pseudomonas strains are on April 10th, 2018 in China typical culture collection
The heart (CCTCC) preservation, deposit number are CCTCC No：M 2018189.
By solid medium culture, it is milk yellow, surface wettability, protrusion, side to observe the entomophila pseudomonad bacterium colony
Edge rounding can generate fluorchrome, pass through micro- sem observation, it was demonstrated that thalline is rod-shaped, can be moved.
It is tested by Physiology and biochemistry, it is a kind of gramnegative bacterium to specify the entomophila pseudomonad, can secrete essence
The double hydrolases of propylhomoserin and protease；Glucose, mannose, mannitol, gluconate, capric acid, malic acid, lemon can be assimilated
Acid, phenylacetic acid；62 kinds of carbon sources including polysorbate40, L-arabinose, D-Fructose, alpha-D-glucose etc. can be utilized.
The entomophila pseudomonad has significant antagonism to xanthomonas oryzae pv. oryzicola.
The xanthomonas oryzae pv. oryzicola includes Xanthomonas oryzae pv. oryzicola (Xanthomonas oryzae
Pv.oryzicola, Xoc RS105) and rice leaf spot bacteria (Xanthomonas oryzae pv.oryzae, Xoo
PXO99A)。
By tablet antagonism method, has detected 3 kinds of the entomophila pseudomonad pair and produce the water for causing economic loss larger
The antagonism of rice bacterial leaf spot pathogenic bacteria, walnut alternaria and ralstonia solanacearum of tomato, find its for rice leaf spot bacteria and
Walnut alternaria also has obvious antagonism.
Further, antagonism of the entomophila pseudomonad to other 8 kinds of cause of disease Xanthomonas campestris is had detected, it was demonstrated that it is to cowpea
Beans wilt, Kidney bean wilt, soybean macula germ and capsicum A. mali also have antagonistic activity.
Tong Guo Dui Chi cultural methods, have detected 2 kinds of disease fungus Pyricularia oryzaes of the entomophila pseudomonad pair and gibberella saubinetii
The antagonism of germ finds that it has apparent antagonistic activity to Pyricularia oryzae.
The second aspect of the present invention is：The separation method of the entomophila pseudomonad is provided：It, will not using dilution coated plate method
It with the soil bacteria suspension of gradient, is coated on the NA tablets of Inoculated Rice Population of Xanthomonas Oryzae Pv bacterial strain, picking can generate apparent suppression
The bacterium colony of bacterium circle, scribing line obtain single bacterium colony, carry out PCR amplification using the universal primer of bacterial 16 S rRNA genes, build system
Chadogram specifies the classification position of the bacterial strain.
The comparative analysis that the present invention passes through 16S rRNA analysis and systematic evolution tree affiliation, it was demonstrated that the bacterial strain is entomophila
Pseudomonad Pseudomonas entomophila, a kind of pseudomonad for capableing of desinsection.
The third aspect of the present invention is：The application of the entomophila pseudomonad is provided, including is applied below：
Application as xanthomonas oryzae pv. oryzicola Antagonistic Fungi；.
Application as rice leaf spot bacteria and/or walnut alternaria Antagonistic Fungi；
Application as cowpea wilt, Kidney bean wilt, soybean macula germ or capsicum A. mali Antagonistic Fungi；
Application as Pyricularia oryzae Antagonistic Fungi.
Compared with prior art, one plant of entomophila pseudomonad 1257 provided by the invention has Xanthomonas oryzae pv. oryzicola aobvious
The antagonism of work, and pathogenetic bacteria to a variety of xanthomonas and Pyricularia oryzae also have antagonistic effect, are that rice is thin
The biological control of fungus diseases provides new resources.
Description of the drawings
Microscope (1000X) the observation photo and colonial morphology of Fig. 1 entomophilas pseudomonad 1257.
Fig. 2 entomophilas pseudomonad 1257 is based on phylogenetic tree gene constructed 16S rRNA.
The Gel electrophoresis results of Fig. 3 .16S rRNA genes.Wherein 1 represents Marker；2 represent CCTCC No：M
The product of 2018189 16S rRNA genes；A is Marker figures, and B is Marker and 16S rRNA gene maps.
Fig. 4 entomophilas pseudomonad 1257 to Xanthomonas oryzae pv. oryzicola (Xanthomonas oryzae pv.oryzicola,
Xoc RS105) antagonistic effect figure.
Fig. 5 entomophilas pseudomonad 1257 is to rice leaf spot bacteria (Xanthomonas oryzae pv.oryzae, Xoo
PXO99A) antagonistic effect figure.
Fig. 6 entomophilas pseudomonad 1257 is to walnut alternaria (Xanthomonas campestris
Pv.juglandis antagonistic effect figure).
Antagonistic effect of Fig. 7 entomophilas pseudomonad 1257 to ralstonia solanacearum of tomato (Ralstonia solanacearum)
Figure.
Fig. 8 entomophilas pseudomonad 1257 is to the antagonistic effect figure of 9 kinds of pathogenic Xanthomonas campestris, A：Xanthomonas oryzae pv. oryzicola
(Xanthomonas oryzae pv.oryzicola,Xoc RS105)；B：Cowpea wilt (Fusarium oxysporum
Schl)；C：Kidney bean wilt (Fusarium oxysporumf sp.phaseoli)；D：Soybean macula germ
(Xanthomonas axonopodis pv.gycines)；E：Capsicum A. mali (Xanthomonas campestris
pv.vesicatoria)；F：Cotton bacterial angular leaf spot bacterium (Xanthomonas campestris pv.malvacearum)；
G：Cobb's disease bacterium (Xanthomonas axonopodis pv.vasculorum)；H：Onion Bacterial Leaf Blight bacterium
(Xanthomonas oampestris pv.cucurbitae)；I：Ralstonia solanacearum (Burkholderia
solanacearum)。
Antagonistic effect figure of Fig. 9 entomophilas pseudomonad 1257 to Pyricularia oryzae (Magnaporthe oryzae), wherein
A：Pyricularia oryzae, B：Pseudomonad 1257 and Pyricularia oryzae Dui Chi culture effect figures.
Antagonistic effect of Figure 10 entomophilas pseudomonad 1257 to fusarium graminearum (Fusarium graminearum)
Figure, wherein A：Fusarium graminearum, B：Pseudomonad 1257 and gibberellic hypha Dui Chi culture effect figures.
Specific implementation mode
It is further illustrated the present invention below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
Product specification selects.It should be appreciated that described herein, specific examples are only used to explain the present invention, without constituting to the present invention
Limitation.
Bacterium culture medium used in following embodiment is as follows：
Beef-protein medium NA (g/L)：Beef extract 3g, polyprotein peptone 5g, sucrose 10g, yeast powder 1g, fine jade
Cosmetics 15g is added water dissolution, is finally settled to 1000mL, adjusts pH 7.0-7.2, high pressure sterilization after packing (121 DEG C,
20min)。
LB culture mediums (g/L)：Tryptone 10g, sodium chloride 10g, Yeast Extract 5g, distilled water 1000mL, pH
7.0-7.2, high pressure sterilization (121 DEG C, 20min).
PDA solid mediums (g/L)：Potato 200g, glucose 20g, agar 15g, distilled water 1000mL, pH 7.0-
7.2, high pressure sterilization (121 DEG C, 10min).
The acquisition of embodiment 1, entomophila pseudomonad 1257
1, soil sources
Senior Residents in Fengxian District of Shanghai vegetable field pakchoi rhizosphere soil
2, the screening of bacterial strain
(1) soil sample acquires
5 point sampling method of zigzag carries out soil sample acquisition：Every acquisition 200g soil sample, will be according to the soil of Z-shaped 5 taken point
Sample is uniformly mixed, and takes 200g to be fitted into sterilizing bag with quartering, as a soil sample.Every piece of ground acquires 3 soil samples as repetition.
Record the time sampled, place and type.The soil sample that acquisition is completed, which is placed in 4 DEG C of refrigerators, to be preserved, in case used in bacterium separation.
(2) separation of bacterium
Using plate dilution method：Each soil sample weighs 10g and puts into sterile triangular pyramidal bottle, and 90mL sterile waters are added,
It is subsequently placed in 200rpm, 15min is vibrated in 28 DEG C of shaking tables, triangular flask is taken out and stands 5min at room temperature, soil bacteria suspension is made
Stoste.Soil bacteria suspension stoste is drawn in centrifuge tube, gradient dilution is carried out, respectively obtains 100、10-1、10-2、10-3、10-4、
10-5、10-6, the bacteria suspension dilution of totally 7 gradients.200 μ L are respectively taken from 7 dilution solution, are spread evenly across containing item
On the NA tablets of pinta bacterium RS105,3 repetitions of each gradient.28 DEG C of biochemical culture carton upside down cultures are placed in, are observed afterwards for 24 hours.
(3) bacteria purification and preservation
There is picking the single bacterium colony of inhibition zone scribing line purifying is carried out on NA tablets, be placed in 28 DEG C of biochemical cultivation case cultures
12h, picking single bacterium colony, number.By inoculation in liquid NA or LB culture medium, 12h is cultivated in 28 DEG C, 200rpm shaking tables
Afterwards, the sterile glycerol for drawing 1mL bacterium solutions and 1mL 50%, gently vibrates mixing, is placed in -80 DEG C of long-term preservations.
(4) screening of Antagonistic Fungi
Using filter paper enzyme：Pathogen is seeded in NA fluid nutrient mediums, 28 DEG C, cultivate 12h in 200rpm shaking tables, draws
After 200 μ L bacteria suspensions and NA solid mediums mix well, a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices is then well-balanced on NA tablets to be stained with a diameter of 7mm's
Filter paper, every filter paper point connect 5 μ L for examination bacterium (OD600 about 2.0), each handle three repetitions be placed in 28 DEG C after drying
Biochemical cultivation case in be inverted culture for 24 hours, seen whether inhibition zone appearance, record strain number simultaneously measures inhibition zone size.
Microscope (1000X) the observation photo and colonial morphology of entomophila pseudomonad 1257 are as shown in Figure 1.
Embodiment 2, the 16S rRNA identified for genes of entomophila pseudomonad 1257
The genomic DNA for extracting bacterial strain 1257, utilizes primer：27F5’-AGAGTTTGATCCTGGCTCAG-3 ' and
1492R5’-TACGGCTACCTTGTTACGACTT-3’；Using the DNA of extraction as template, PCR amplification is carried out, to obtain purpose piece
Section.PCR reaction systems are：
Reaction mixture gently of short duration centrifugation after mixing, that is, carry out PCR amplification.PCR reaction conditions are：
94 DEG C of 3min,
94 DEG C of 30sec, 56 DEG C of 30sec, 72 DEG C of 90sec, 30 cycles,
72 DEG C of 8min,
25℃Forever。
Take 5 μ L of above-mentioned pcr amplification product, using 1% Ago-Gel carry out electrophoresis detection after (see Fig. 3), by PCR original
Ye Songboshang biotechnologys (Shanghai) Co., Ltd. is sequenced, and base sequence is as shown in SEQ ID No.1.
Sequencing result is analyzed with DNA Star, and BLAST comparisons are carried out on the websites NCBI, determines that nearly edge strain is thin
The kind of bacterium.
As a result it shows：Bacterial strain 1257 and Pseudomonas entomophila have 99.4% similarity.It uses
MEGA6.0 phylogenetic tree constructions, are as a result shown in Fig. 2.
The Physiology and biochemistry of embodiment 3,1257 is identified
The physiological and biochemical property of entomophila pseudomonad 1257 of the present invention is：Fluorchrome can be generated, essence can be secreted
The double hydrolases of propylhomoserin and protease, can assimilate glucose, mannose, mannitol, gluconate, capric acid, malic acid, lemon
Acid, phenylacetic acid, are shown in Table 1.62 kinds of carbon sources such as polysorbate40, L-arabinose, D-Fructose, alpha-D-glucose can be utilized.It is unable to water
Indoles, aesculin, beta galactoside enzyme are solved, nitrate reductase to nitrite cannot be shown in Table 2.
1 1257 physio-biochemical characteristics of table-enzyme activity, carbon assimilation
+：Positive reaction；-：Negative reaction；
The physio-biochemical characteristics-of table 2 1257 produce acid using carbon source
+：Positive reaction；-：Negative reaction；W：Weakly positive is reacted
Embodiment 4, the antagonism spectrum of entomophila pseudomonad 1257 measure
1) entomophila pseudomonad 1257 measures the antagonistic activity of 12 kinds of phytopathogens
By Xanthomonas oryzae pv. oryzicola, rice leaf spot bacteria, walnut alternaria, ralstonia solanacearum of tomato and xanthomonas
8 kinds of pathogens and be inoculated in respectively in NA fluid nutrient mediums for examination pseudomonad 1257,28 DEG C, cultivate in 200rpm shaking tables
After 12h, unified OD600It is 2.0；It draws respectively after the corresponding cause of disease bacterium solutions of 200 μ L and NA solid mediums mix well,
Plate, the then filter paper for being stained with a diameter of 7mm well-balanced on NA tablets, every filter paper point connect 5 μ L for trying pseudomonad
1257, three repetitions are each handled, after drying, is placed in 28 DEG C of biochemical cultivation case and is inverted culture for 24 hours, seen whether suppression
Bacterium is irised out now, and records inhibition zone size.
Fungistatic effect of the 3 entomophila pseudomonad 1257 of table to Different Kinds of Pathogens bacterium
Entomophila pseudomonad 1257 is to Xanthomonas oryzae pv. oryzicola (Xanthomonas oryzae pv.oryzicola, Xoc
RS105 antagonistic effect figure) as shown in figure 4, entomophila pseudomonad 1257 to rice leaf spot bacteria (Xanthomonas
oryzae pv.oryzae,Xoo PXO99A) antagonistic effect figure as shown in figure 5, entomophila pseudomonad 1257 to walnut blackspot
The antagonistic effect figure of germ (Xanthomonas campestris pv.juglandis) is as shown in Figure 6.Entomophila pseudomonad
The antagonistic effect figure of 1257 pairs of ralstonia solanacearum of tomato (Ralstonia solanacearum) is as shown in Figure 7.Entomophila pseudomonad
The antagonistic effect figures of 1257 pairs of 9 kinds of pathogenic Xanthomonas campestris is as shown in figure 8, in Fig. 8, A：Xanthomonas oryzae pv. oryzicola
(Xanthomonas oryzae pv.oryzicola,Xoc RS105)；B：Cowpea wilt (Fusarium oxysporum
Schl)；C：Kidney bean wilt (Fusarium oxysporumf sp.phaseoli)；D：Soybean macula germ
(Xanthomonas axonopodis pv.gycines)；E：Capsicum A. mali (Xanthomonas campestris
pv.vesicatoria)；F：Cotton bacterial angular leaf spot bacterium (Xanthomonas campestris pv.malvacearum)；
G：Cobb's disease bacterium (Xanthomonas axonopodis pv.vasculorum)；H：Onion Bacterial Leaf Blight bacterium
(Xanthomonas oampestris pv.cucurbitae)；I：Ralstonia solanacearum (Burkholderia
solanacearum)。
2) entomophila pseudomonad 1257 measures the antagonism of fungal disease
Using tablet opposite culture method, fusarium graminearum and rice blast fungus are cultivated in PDA culture medium respectively,
It after pathogen covers with tablet, being beaten in plate edge with card punch and takes fungus block spare, take one of bacteria cake, mycelia is face-down,
It is inoculated into new PDA plate center, is stained with the filter paper of same size in bacteria cake surrounding equidistant (25mm) using crossing method
Piece (7mm), on every filter paper point connect 5 μ L for trying pseudomonad 1257, each handle 3 repetitions, meanwhile, it is false single not connect
The tablet that born of the same parents bacterium only connects disease fungus is control.Inversion culture is carried out in 28 DEG C of biochemical cultivation cases, and suppression is observed and recorded after 5 days
Bacterium phenomenon, antibacterial situation such as attached drawing 9 and attached drawing 10.The results show that pseudomonad 1257 has certain suppression to rice blast
Effect processed.
One plant of entomophila pseudomonad 1257 provided by the invention is utilized as a result, is had to Xanthomonas oryzae pv. oryzicola significant short of money
Anti- effect, and pathogenetic bacteria to a variety of xanthomonas and Pyricularia oryzae also have antagonistic effect.For rice bacteriosis
Biological control provide new resources.
The above description of the embodiments is intended to facilitate ordinary skill in the art to understand and use the invention.
Person skilled in the art obviously easily can make various modifications to these embodiments, and described herein general
Principle is applied in other embodiment without having to go through creative labor.Therefore, the present invention is not limited to the above embodiments, ability
Field technique personnel announcement according to the present invention, improvement and modification made without departing from the scope of the present invention all should be the present invention's
Within protection domain.
Sequence table
<110>Shanghai Communications University
<120>One pseudomonas and its separation method and application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1416
<212> DNA
<213>Entomophila pseudomonad (Pseudomonas entomophila)
<400> 1
ctacacatgc aagtcgagcg gatgagaaga gcttgctctt cgattcagcg gcggacgggt 60
gagtaatgcc taggaatctg cctggtagtg ggggacaacg tttcgaaagg aacgctaata 120
ccgcatacgt cctacgggag aaagcagggg accttcgggc cttgcgctat cagatgagcc 180
taggtcggat tagctagttg gtgaggtaat ggctcaccaa ggcgacgatc cgtaactggt 240
ctgagaggat gatcagtcac actggaactg agacacggtc cagactccta cgggaggcag 300
cagtggggaa tattggacaa tgggcgaaag cctgatccag ccatgccgcg tgtgtgaaga 360
aggtcttcgg attgtaaagc actttaagtt gggaggaagg gcagtaagtt aataccttgc 420
tgttttgacg ttaccgacag aataagcacc ggctaactct gtgccagcag ccgcggtaat 480
acagagggtg caagcgttaa tcggaattac tgggcgtaaa gcgcgcgtag gtggttcgtt 540
aagttggatg tgaaagcccc gggctcaacc tgggaactgc atccaaaact ggcgagctag 600
agtatggtag agggtggtgg aatttcctgt gtagcggtga aatgcgtaga tataggaagg 660
aacaccagtg gcgaaggcga ccacctggac tgatactgac actgaggtgc gaaagcgtgg 720
ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgtca actagccgtt 780
ggaatccttg agattttagt ggcgcagcta acgcattaag ttgaccgcct ggggagtacg 840
gccgcaaggt taaaactcaa atgaattgac gggggcccgc acaagcggtg gagcatgtgg 900
tttaattcga agcaacgcga agaaccttac caggccttga catgcagaga actttccaga 960
gatggattgg tgccttcggg aactctgaca caggtgctgc atggctgtcg tcagctcgtg 1020
tcgtgagatg ttgggttaag tcccgtaacg agcgcaaccc ttgtccttag ttaccagcac 1080
gttatggtgg gcactctaag gagactgccg gtgacaaacc ggaggaaggt ggggatgacg 1140
tcaagtcatc atggccctta cggcctgggc tacacacgtg ctacaatggt cggtacagag 1200
ggttgccaag ccgcgaggtg gagctaatct cacaaaaccg atcgtagtcc ggatcgcagt 1260
ctgcaactcg actgcgtgaa gtcggaatcg ctagtaatcg caaatcagaa tgttgcggtg 1320
aatacgttcc cgggccttgt acacaccgcc cgtcacacca tgggagtggg ttgcaccaga 1380
agtagctagt ctaaccttcg ggaggacggt accacg 1416
Claims (10)
1. one plant of entomophila pseudomonad, which is characterized in that the pseudomonas strains are trained on April 10th, 2018 in Chinese Typical Representative
Object collection preservation is supported, deposit number is CCTCC No：M 2018189.
2. entomophila pseudomonad according to claim 1, which is characterized in that the entomophila pseudomonad bacterium colony is milk yellow,
Surface wettability, protrusion, edge rounding can generate fluorchrome, and thalline is rod-shaped, can be moved.
3. entomophila pseudomonad according to claim 1, which is characterized in that the entomophila pseudomonad is a kind of gram-negative
Property bacterium, can secrete arginine dihydrolase and protease；Glucose, mannose, mannitol, gluconic acid can be assimilated
Salt, capric acid, malic acid, citric acid, phenylacetic acid；It includes polysorbate40, L-arabinose, D-Fructose, alpha-D-glucose that can utilize
Carbon source inside.
4. entomophila pseudomonad according to claim 1, which is characterized in that the entomophila pseudomonad is to paddy bacterial item
Pinta bacterium has antagonism.
5. entomophila pseudomonad according to claim 4, which is characterized in that the xanthomonas oryzae pv. oryzicola includes rice
Population of Xanthomonas Oryzae Pv (Xanthomonas oryzae pv.oryzicola, Xoc RS105) and rice leaf spot bacteria
(Xanthomonas oryzae pv.oryzae,Xoo PXO99A)。
6. entomophila pseudomonad according to claim 1, which is characterized in that the entomophila pseudomonad is to bacterial blight of rice
Bacterium and walnut alternaria have antagonism.
7. entomophila pseudomonad according to claim 1, which is characterized in that the entomophila pseudomonad is to cowpea droop
Bacterium, Kidney bean wilt, soybean macula germ and capsicum A. mali have antagonistic activity.
8. entomophila pseudomonad according to claim 1, which is characterized in that the entomophila pseudomonad has Pyricularia oryzae
Antagonistic activity.
9. the separation method of entomophila pseudomonad as described in claim 1, which is characterized in that include the following steps：
Using dilution coated plate method, by the soil bacteria suspension of different gradients, the NA for being coated on Inoculated Rice Population of Xanthomonas Oryzae Pv bacterial strain is flat
On plate, picking can generate the bacterium colony of apparent inhibition zone, and scribing line obtains single bacterium colony, utilizes general the drawing of bacterial 16 S rRNA genes
Object carries out PCR amplification, builds systematic evolution tree, specifies the classification position of the bacterial strain.
10. the application of entomophila pseudomonad as described in claim 1, which is characterized in that including applying below：
Application as xanthomonas oryzae pv. oryzicola Antagonistic Fungi；
Application as rice leaf spot bacteria and/or walnut alternaria Antagonistic Fungi；
Application as cowpea wilt, Kidney bean wilt, soybean macula germ or capsicum A. mali Antagonistic Fungi；
Application as Pyricularia oryzae Antagonistic Fungi.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112111426A (en) * | 2020-09-22 | 2020-12-22 | 河南科技学院 | Pseudomonas flaxseed and application thereof in biocontrol of green Chinese onion purple spot |
| CN112175869A (en) * | 2020-09-29 | 2021-01-05 | 烟台市林业科学研究所 | Pseudomonas and application thereof |
| CN112300964A (en) * | 2020-11-04 | 2021-02-02 | 福建农林大学 | Insect pseudomonas with insecticidal activity on coleoptera pests and application thereof |
| CN112359000A (en) * | 2020-11-27 | 2021-02-12 | 江苏省农业科学院 | Efficient biocontrol pseudomonas and application thereof in prevention and control of rice diseases |
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| US6048713A (en) * | 1995-03-28 | 2000-04-11 | Nisshin Flour Milling Co., Ltd. | Pseudomonas fluorescens |
| CN105925501A (en) * | 2016-05-09 | 2016-09-07 | 中国水稻研究所 | Pseudomonas mosselii and application thereof |
| CN106978373A (en) * | 2017-05-09 | 2017-07-25 | 福建农林大学 | One plant is used for pseudomonad and the application that bacteriosis is prevented and treated |
| CN107502584A (en) * | 2017-10-16 | 2017-12-22 | 山东省林业科学研究院 | One fluorescent pseudomonads and its application in walnut black spot is prevented and treated |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US6048713A (en) * | 1995-03-28 | 2000-04-11 | Nisshin Flour Milling Co., Ltd. | Pseudomonas fluorescens |
| CN105925501A (en) * | 2016-05-09 | 2016-09-07 | 中国水稻研究所 | Pseudomonas mosselii and application thereof |
| CN106978373A (en) * | 2017-05-09 | 2017-07-25 | 福建农林大学 | One plant is used for pseudomonad and the application that bacteriosis is prevented and treated |
| CN107502584A (en) * | 2017-10-16 | 2017-12-22 | 山东省林业科学研究院 | One fluorescent pseudomonads and its application in walnut black spot is prevented and treated |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112111426A (en) * | 2020-09-22 | 2020-12-22 | 河南科技学院 | Pseudomonas flaxseed and application thereof in biocontrol of green Chinese onion purple spot |
| CN112111426B (en) * | 2020-09-22 | 2022-10-21 | 河南科技学院 | Pseudomonas flaxseed and application thereof in biocontrol of green Chinese onion purple spot |
| CN112175869A (en) * | 2020-09-29 | 2021-01-05 | 烟台市林业科学研究所 | Pseudomonas and application thereof |
| CN112300964A (en) * | 2020-11-04 | 2021-02-02 | 福建农林大学 | Insect pseudomonas with insecticidal activity on coleoptera pests and application thereof |
| CN112359000A (en) * | 2020-11-27 | 2021-02-12 | 江苏省农业科学院 | Efficient biocontrol pseudomonas and application thereof in prevention and control of rice diseases |
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| Publication number | Publication date |
|---|---|
| CN108441456B (en) | 2020-08-25 |
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