CN102191201B - Pine endosymbiotic bacterium-new burkholderia cenocepacia NSM-05 and application thereof - Google Patents
Pine endosymbiotic bacterium-new burkholderia cenocepacia NSM-05 and application thereof Download PDFInfo
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Abstract
The invention discloses a pine endosymbiotic bacterium which is classified and named after a new burkholderia cenocepacia NSM-05 and is stored in the China typical culture collection center with the storage number of CCTCC NO: M2011015 and the storage data of January 11, 2011. The invention also discloses application of the new burkholderia cenocepacia NSM-05 to the preventing and treating pine shoot dieback diseases and promoting the growth of pines. The invention also discloses an antagonistic inoculant for preventing and treating the pine shoot dieback diseases; the antagonistic inoculant is obtained by inoculating NSM-05 bacterium strains in a culture medium. The NSM-05 bacterium strain provided by the invention is the pine endosymbiotic bacterium, has strong bacteriostasis on the pine shoot dieback diseases, has the antagonism effect on multiple woods and crop pathogenic bacteria and road-spectrum antibacterial activity, is safe and non-toxic to people, livestock and plants and has good development and application prospects.
Description
Technical field
The present invention relates to the endogenetic bacteria of plant, be specifically related to a kind of pine tree endogenetic bacteria new onion burkholderia NSM-05 and application thereof.
Background technology
Pine tree shoot blight (Sphaeropsis sapinea) is the most common and one of the widest important disease that distributes on the interior softwood tree of world wide.In nearly 30 countries that the whole world should disease distributes, the most serious country that is injured has: nz, Australia, South Africa, Holland, the U.S. and China.Its host range is very wide, can infect more than 60 kind of softwood tree of Pinus (Pinus L.), abies (Abies Mill), Larix (Larix Mill), Thuja-Arten (Thuja L.), Cedrus (Cedrus Trew), Juniperus Linn. (Juniperus L.), Picea (Picea Dietr.) and Pseudotsuga 8 genus such as (Pseudotsuga Carr).Though the host range of pine tree shoot blight pathogenic bacteria is wide, the susceptible degree of different host is different, at present known cause financial loss slash pine, torch pine, Pinus massoniana Lamb and pinus sylvestris var. mongolica only arranged.Since nineteen eighty; Take place in the external pine forest in more than 10 provinces such as this disease is Central-South, southwestern in China, south China, East China, northwest; Caused Direct Loss are mainly due to the withered high growth retardation of pine tree that causes of leading shoot; The side tip is grown thickly and volume increment is fallen sharply, when serious even make whole strain pine tree withered; Its indirect loss main with the timber that timber blue stain etc. is caused demote relevant, thereby the commodity value of timber is greatly reduced.
Endophytic bacterium is meant and has in the plant materials bacterioid of plant not being seen harm, can grow surely in vegetable cell gap or cell, and set up harmony with host plant and unite relation.Since endogenetic bacteria can system be distributed in the plant materials; Competent nitrogenous source and carbon source are arranged, have good living environment, overcome in the past from PGPR (plant growth pro-moting rhizobacteria; Rhizosphere is urged endophytic bacteria) in the screening biocontrol microorganisms shortcoming (it is poor to grow surely; Affected by environment big), so more can bring into play its biocontrol effect, have broad application prospects.And find at present to utilize the pine tree endogenetic bacteria to prevent and treat the relevant report of sphaeropsis sapinea.
New onion burkholderia is under the jurisdiction of the onion bulkholderia cepasea at present on classification position.Onion Burkholderia (Burkholderia cepacia; Be called for short Bc); Being a kind of gram negative bacterium that extensively is present in soil, water, plant surface and hospital, is the mixture that one group of genotype is different, phenotype is close (Burkholderia cepacia complex is called for short Bcc); Be divided into 10 genotype now in the world, close and claim onion Burkholderia crowd (Bcc).Many research shows that some genotype of Bc have multiple distinctive effect, and it can decompose agricultural chemicals residual in the soil, purify waste water; It can controlling plant diseases, and as biocontrol microorganisms, Bc is made into biotic pesticide and biological bactericide, can substitute part chemical pesticide commonly used, is playing important effect aspect agriculture prodn and the environment protection; It can also promote the growth of plant, in agriculture prodn as tethelin and compound manure.It is the raw material production biological pesticide that existing at present many countries adopt Bc, but does not also appear in the newspapers with Bc control sphaeropsis sapinea.
Owing to increasing of using of microbial pesticide and some biocontrol microorganisms its risk that possibly become the human body conditioned pathogen that often has been left in the basket is registered use, the risk assessment of microbial pesticide becomes the problem that the whole world is paid close attention to day by day.The security that the plant-animal model of at present more existing comparative maturities is used for the biological pesticide bacterial strain detects, and assesses these biocontrol microorganisms whether people and animals' plant environment is existed potential risk.
Summary of the invention
Goal of the invention: the deficiency to existing in the prior art the purpose of this invention is to provide the new onion burkholderia of a kind of pine tree endogenetic bacteria NSM-05, to realize with it pine tree shoot blight being carried out biological control.
Another object of the present invention provides the purposes of above-mentioned new onion burkholderia NSM-05.
Technical scheme: in order to realize the foregoing invention purpose, the technical scheme that the present invention adopts is: a kind of pine tree endogenetic bacteria, its classification called after onion burkholderia (
Burkholderia cenocepacia) NSM-05, being preserved in Chinese typical culture collection center, its deposit number is: CCTCC NO:M 2011015, preservation date: on January 11st, 2011.
New onion burkholderia NSM-05 bacterial strain; What be that separation screening obtains from the healthy Pinus massoniana Lamb branch has good biocontrol effect and the endogenetic bacteria that promotes growth to sphaeropsis sapinea; This endogenetic bacteria can well be grown in the pine tree body surely; To people and animals and plant safety non-toxic, also have following character:
1, morphological specificity
Circular, the point-like of bacterium colony on the beef-protein medium flat board, faint yellow, the edge is complete, smooth surface, protuberance; Shaft-like, the no gemma of thalline, size 2.8 ~ 5.2 * 0.9 ~ 2.8 μ m, aerobic, oxidase test is positive, Gram-negative, 3%KOH tests the positive.
2,16S rDNA analyzes
Record 16SrDNA sequence 1302bp, specifically shown in SEQ ID No.1.The 16SrDNA sequence finds the very high close bacterial strain sequence of homology through the BLAST comparison in GenBank.With NSM-05 bacterial strain similarity the highest be new onion burkholderia (
Burkholderia cenocepacia), homology reaches 99%.Through the BIOLOG system NSM-05 is identified, this bacterial strain identification plate cultivate 16～24h and new onion burkholderia (
Burkholderia cenocepacia) reading similar value (SIM value) is 0.545, greater than 0.50, meets the requirement of BIOLOG system about desired result, so this bacterial strain utilizes the BIOLOG system, accurately identified kind, for new onion burkholderia (
Burkholderia cenocepacia).
The application of above-mentioned pine tree endogenetic bacteria in the biological control of pine tree shoot blight.New onion burkholderia NSM-05 to sphaeropsis sapinea bacterium Sphaeropsis sapinea (
Sphaeropsis sapinea) have an intensive bacteriostatic action.
The application of above-mentioned pine tree endogenetic bacteria in promoting the pine tree growth.New onion burkholderia NSM-05 is good to pine tree shoot blight field biological and ecological methods to prevent plant disease, pests, and erosion action effect, and above-mentioned new onion burkholderia NSM-05 also has the growth of promotion effect to pine tree simultaneously.
The application of above-mentioned pine tree endogenetic bacteria in suppressing the forest pathogenic bacteria, promptly new onion burkholderia NSM-05 is to 6 kinds of forest pathogenic bacterias: Ginkgo Leaf rot alternaric bacteria (
Alternaria alternata), Liriodendron black spot interlinkage spore (
Alternaria sp.), Liriodendron black spot anthrax-bacilus (
Colletotrichum sp.), the dothiorella gregaria bacterium: Phomopsis (
Phomopsis sp.), golden yellow gram capsule spore (
Cytospora chrysosperma), horse-chestnut shell shuttle spore (
Fusicoccum aesculi) have bacteriostatic action, can be applicable to prevent and treat Ginkgo Leaf rot, Liriodendron black spot, dothiorella gregaria.
The application of above-mentioned pine tree endogenetic bacteria in suppressing pathogen of crop, promptly new onion burkholderia NSM-05 is to 7 kinds of pathogen of crop: Rhizoctonia solani Kuhn (
Thanatephorus cucumeris), fusarium graminearum (
Gibberella zeae), cotton rhizoctonia solani (
Rhizoctonia solani), cotton-wilt fusarium (
Fusarium oxysoorium), the ring rot of apple bacterium (
Dothiorella gregaria), tea moire blight bacterium (
Guignardia camelliae), tea zonate spot bacterium (
Pestalotiopsis versicolor) have bacteriostatic action, can be applicable to prevent and treat rice sheath blight disease, wheat scab, cotton seedling blight, cotton wilt, ring rot of apple, tea moire blight, tea zonate spot.
A kind of sphaeropsis sapinea biocontrol fungicide adopts following method preparation: new onion burkholderia NSM-05 is inoculated in substratum, air flow 1:1; 28 ~ 30 ℃ of temperature; Shaking table revolution 180 ~ 200r/min, 48 ~ 60h, the fermented liquid that obtains is the sphaeropsis sapinea biocontrol fungicide; Wherein, the prescription of substratum is: in every 1000ml zero(ppm) water, contain Carnis Bovis seu Bubali cream 2 ~ 5g, peptone 5 ~ 10g, sodium-chlor 5g, pH5 ~ 7.
This fermented liquid directly sprays application to the pine tree tip of a branch and limb going up just can be brought into play its biological control effect.
Beneficial effect: a kind of new onion burkholderia NSM-05 bacterial strain provided by the invention is a kind of pine tree endogenetic bacteria, and strong to sphaeropsis sapinea bacterium bacteriostatic action, biological and ecological methods to prevent plant disease, pests, and erosion test effect in field is good.Pine tree also had the growth of promotion effect.And multiple forest and pathogen of crop are had bacteriostatic action, have antimicrobial spectrum very widely.Culture condition is simple, is easy to preserve and produce, and colonization ability is strong in the pine tree body; Fast growth; Overcome in the past biocontrol microorganisms and grow shortcomings such as poor, biocontrol effect instability surely, in biocontrol of plant disease, play important effect; Particularly the control to sphaeropsis sapinea has great importance, and is a kind of broad spectrum type sterilant with good development prospect.
Description of drawings
Fig. 1 is new onion burkholderia NSM-05 bacterial strain to the antagonistic action of sphaeropsis sapinea bacterium figure as a result.Wherein, A figure is new onion burkholderia NSM-05 bacterial strain, and B figure is contrast (CK);
Fig. 2 is new onion burkholderia NSM-05 meta-bolites to the antagonistic action of sphaeropsis sapinea bacterium 2d figure as a result.Wherein, A figure is new onion burkholderia NSM-05 meta-bolites, and B figure is CK;
Fig. 3 is new onion burkholderia NSM-05 meta-bolites to the antagonistic action of sphaeropsis sapinea bacterium 5d figure as a result.Wherein, A figure is new onion burkholderia NSM-05 meta-bolites, and B figure is CK;
Fig. 4 is new onion burkholderia NSM-05 to sphaeropsis sapinea field control figure as a result.Wherein, left side figure is not biological control, and right figure adopts new onion burkholderia NSM-05 that sphaeropsis sapinea is carried out biological control;
Fig. 5 is new onion burkholderia NSM-05 to the growth promoting function of slash pine figure as a result.Wherein, left side figure is for being untreated, and right figure adopts new onion burkholderia NSM-05 to handle;
Fig. 6 is new onion burkholderia NSM-05 to the pathogenic mensuration of onion figure as a result.Among the figure, A is the onion of inoculation sterilized water, and B is the onion of inoculation onion burkholderia LMG1222.C is the onion of the new onion burkholderia NSM-05 of inoculation;
Fig. 7 is new onion burkholderia NSM-05 to the pathogenic mensuration of onion exploded chart as a result.Wherein, A figure is the onion of inoculation sterilized water, and B figure is the onion of inoculation onion burkholderia LMG1222.C figure is the onion of the new onion burkholderia NSM-05 of inoculation;
Fig. 8 is new onion burkholderia NSM-05 to the pathogenic mensuration of clover figure as a result.Among the figure, A is the clover of inoculation sterilized water, and B is the clover of inoculation onion burkholderia LMG1222, and C is the clover of the new onion burkholderia NSM-05 of inoculation;
Fig. 9 is the bacteriostatic activity table of new onion burkholderia NSM-05 meta-bolites to the sphaeropsis sapinea bacterium.
Embodiment
Below in conjunction with specific embodiment the present invention is done further explanation.
The employed culture medium prescription of following examples is:
PDA plate culture medium prescription: yam 200g, sucrose 20g, agar 20g, zero(ppm) water 1L;
The NA culture medium prescription: Carnis Bovis seu Bubali cream 3g, peptone 10g, sodium-chlor 5g, agar 20g, pH 7.3, zero(ppm) water 1L;
The NB culture medium prescription: Carnis Bovis seu Bubali cream 3g, peptone 10g, sodium-chlor 5g, agar 20g, pH 7.3, zero(ppm) water 1L.
Embodiment 1 new onion burkholderia NSM-05 to the sphaeropsis sapinea bacterium (
Sphaeropsis sapinea) bacteriostatic action.
Adopt the PDA plate culture medium, 25 ℃, constant temperature culture sphaeropsis sapinea bacterium 3 ~ 4 days; Beat the bacterium piece of getting the same amount of homogeneity with the punch tool of diameter 5mm, be inoculated in the dull and stereotyped central authorities of PDA, use the good new onion burkholderia NSM-05 streak inoculation of NA medium slant activation again in both sides apart from sphaeropsis sapinea bacterium bacterium piece 3.5cm; Dull and stereotyped with the PDA that only inoculates the sphaeropsis sapinea bacterium as contrast; Each handles 3 repetitions, and 28 ℃ of constant temperature culture 3 ~ 5 days treat that contrast covers with whole observations when dull and stereotyped.Test-results is as shown in Figure 1, and new onion burkholderia NSM-05 has the intensive bacteriostatic action to the sphaeropsis sapinea bacterium, and inhibition zone (distance of pathogenic bacteria and NSM-05 bacterial strain) diameter MV is 11.15mm.
The new onion burkholderia NSM-05 of inoculation in the NB nutrient solution, 30 ℃, 200r/min shakes training 48h, makes fermented liquid.Then with fermented liquid at 4 ℃, 10000r/min, centrifugal 20 minutes.Get supernatant,, obtain not having the meta-bolites that fermented liquid is the NSM-05 bacterial strain through the biofilter filtration sterilization of 0.22 μ m micropore.The PDA medium sterilization for preparing is cooled to about 50 ℃, and the substratum of every 10ml adds 1ml does not have flat board of fermented liquid mixing, and it is dull and stereotyped to process the band poison.Inoculation sphaeropsis sapinea bacterium after 2 days, and with the PDA culture medium inoculated sphaeropsis sapinea bacterium that do not add no fermented liquid as contrast.Cultivate 5d for 25 ℃, observe its growing state, calculate bacteriostasis rate.Test-results such as Fig. 9, Fig. 2 and shown in Figure 3, no fermented liquid has tangible antagonistic action to the sphaeropsis sapinea bacterium, and the bacteriostasis rate of 2d is 80.18%, and the bacteriostasis rate 57% of 5d explains that new onion burkholderia NSM-05 meta-bolites has antibacterial substance.The calculation formula of bacteriostasis rate is: growth bacteriostasis rate (%)=(contrast flat-plate bacterial colony diameter-be with malicious flat-plate bacterial colony diameter) ÷ (contrast flat-plate bacterial colony diameter-bacterium piece diameter) * 100%.
Embodiment 3 new onion burkholderia NSM-05 are to the sphaeropsis sapinea field control.
5 ~ June in 2010, pineland carries out new onion burkholderia NSM-05 to the sphaeropsis sapinea field control in mountain forest pathological study institute of Nanjing University of Nanjing Forestry University.The inoculation method of sphaeropsis sapinea bacterium: earlier gently slash pine tip of a branch limb is scratched the epidermis of 1cm left and right sides length with the aseptic inoculation cutter, again with sphaeropsis sapinea bacterium fragmentation liquid spray inoculation in the slash pine tip of a branch.Handle 1: 3d irritates root and inoculates new onion burkholderia NSM-05 bacterial strain fermentation liquor (embodiment 2 preparations) 100ml/ strain before the inoculation sphaeropsis sapinea bacterium.Handle 2: the new onion burkholderia of 3d spray inoculation NSM-05 bacterial strain fermentation liquor is till pine tree is drenched before the inoculation sphaeropsis sapinea bacterium.Handle 3: direct inoculation sphaeropsis sapinea bacterium.Each handles 20 strains, 3 repetitions.Observe incidence every day.Inoculation 10d " Invest, Then Investigate " incidence.Statistics disease index and protection effect.The morbidity grade scale is as shown in table 2, test-results as as table 3 with shown in Figure 4, newly onion burkholderia NSM-05 has good biological and ecological methods to prevent plant disease, pests, and erosion effect to sphaeropsis sapinea.
Table 2 sphaeropsis sapinea inoculation morbidity grade scale
| Sick level | Grade scale | Typical value |
| Ⅰ | The inoculation wound healing, no needle yellow | 0 |
| Ⅱ | Near inoculation mouthful 2-3 bundle needle yellow | 1 |
| Ⅲ | The blue brown shrinkage of inoculation wound branch, more needle yellow | 2 |
| Ⅳ | The whole tip of a branch is withered | 3 |
The new onion burkholderia NSM-05 of table 3 is to the field control effect of sphaeropsis sapinea
| Sample | Disease index (%) | Control effect (%) |
| Handle 1 | 6．67 | 87.49 |
| |
3.33 | 93.76 |
| Handle 3 | 53.33 | / |
Wherein, the calculation formula of disease index is: disease index (%)=∑ (sick tree strain tree * typical value at different levels) ÷ (highest typical value is set * fallen ill in total strain) * 100%.The calculation formula of control effect is: control effect (%)=(disease index after contrast disease index-control) ÷ contrasts disease index * 100%.
Embodiment 4 new onion burkholderia NSM-05 are to the growth promoting function of slash pine.
Irritate two kinds of methods of root and spraying with new onion burkholderia NSM-05 fermented liquid (embodiment 2 preparations) and inoculate the potted plant seedling of annual slash pine, compare with the loose seedling that does not have inoculation, each handles 30 strains, surveys its plant height leading thread after 6 months.Test-results such as table 4 are with shown in Figure 5, and new onion burkholderia NSM-05 fermented liquid has tangible growth promoting function to the slash pine seedling, no matter are that the slash pine seedling of irritating after root or spray inoculation are handled exists significant difference with contrast on plant height and leading thread.
Table 4 pine tree endogenetic bacteria NSM-05 is to the growth promoting function of slash pine
| Handle | Plant height (cm) | Leading thread (mm) |
| Irritate root | 29.55a | 3.18a |
| Spraying | 28.86a | 3.15a |
| CK | 23.71b | 2.50b |
Annotate: P 0.05, same column different rows lowercase representative significant difference inequality.
Embodiment 5 new onion burkholderia NSM-05 are to the bacteriostatic action of 6 kinds of forest pathogenic bacterias.
Adopt dull and stereotyped face-off method to detect new onion burkholderia NSM-05 to Ginkgo Leaf rot alternaric bacteria, Liriodendron black spot interlinkage spore, Liriodendron black spot anthrax-bacilus, dothiorella gregaria bacterium: the bacteriostatic action of 6 kinds of forest pathogenic bacterias such as Phomopsis, golden yellow gram capsule spore, horse-chestnut shell shuttle spore.Test-results shows that new onion burkholderia NSM-05 all has certain antagonistic action to whole forest pathogenic bacterias, and antibacterial circle diameter is all more than 5mm.
Embodiment 6 new onion burkholderia NSM-05 are to the bacteriostatic action of 7 kinds of pathogen of crop.
Adopt dull and stereotyped face-off method to detect the bacteriostatic action of new onion burkholderia NSM-05 to 7 kinds of pathogen of crop such as Rhizoctonia solani Kuhn, fusarium graminearum, cotton rhizoctonia solani, cotton-wilt fusarium, ring rot of apple bacterium, tea moire blight bacterium, tea zonate spot bacterium.Test shows; New onion burkholderia NSM-05 all has certain antagonistic action to whole pathogen of crop; All more than 5mm, particularly the bacteriostatic diameter to cotton rhizoctonia solani, cotton-wilt fusarium, ring rot of apple bacterium and tea moire blight bacterium reaches more than the 10mm antibacterial circle diameter.Above-mentioned test shows that new onion burkholderia NSM-05 has antibacterial broad spectrum.
The endogenous mensuration of embodiment 7 new onion burkholderia NSM-05.
The new onion burkholderia NSM-05 that adopts Rifampin (300 mcg/ml) mark to cross is inoculated in the NB substratum, fermentation 36h, and the fermented liquid that obtains sprays in slash pine, Pinus massoniana Lamb, black pine needle and limb.In 2d, 3d, 5d, 7d, 10d, 15d sampling, get needle, limb and Gen Ge 0.4g, surface sterilization respectively; Grind with sterilized water; Whether coat on the NA flat board of 300 μ g/mL, cultivate 5d for 30 ℃, observing flat board has new onion burkholderia NSM-05 bacterium colony to grow.Test shows that slash pine, Pinus massoniana Lamb, black pine limb and root can reclaim new onion burkholderia NSM-05, and it has endogeny.
Embodiment 8 new onion burkholderia NSM-05 are to the pathogenic mensuration of onion.
New onion burkholderia NSM-05, onion burkholderia LMG1222 (being provided by professor Swings of DSMZ of country of Ghent, Belgium university) reference culture shake training 24h at 30 ℃ under the 200r/min condition, the fermented liquid that obtains is centrifugal; 10000r/min, centrifugal 10min abandons supernatant; Add the sterilized water mixing, continue centrifugal under similarity condition, 3-5 time so repeatedly; Metabolite removal until bacterium is clean, obtains pure bacteria suspension, regulates cell concentration 1 * 10
8About cfu/ml.With syringe get respectively be ready to new onion burkholderia NSM-05, the pure bacteria suspension of onion burkholderia LMG1222 reference culture and sterilized water on onion bulb stem two point symmetries injections (degree of depth is 1 ~ 2cm); Wherein reference culture LMG1222 is as positive control, and sterilized water is as negative control.Every some injection volume 1ml, all processing all repeat 4 times, place 25 ℃ of incubated at room temperature, observations behind the 5d.
Test-results such as Fig. 6 and shown in Figure 7 behind the onion bulb stem inoculation 5d; Do not cause behind the new onion burkholderia of the injection inoculation NSM-055d that onion bulb stem rots, only produce very little necrotic spot, continue to cultivate at inoculation point epidermis; Scab does not continue to enlarge; After the onion incision, inside does not have ANOMALOUS VARIATIONS, does not have notable difference with the sterilized water contrast; And positive control reference culture onion burkholderia LMG1222 produces the scab of bulk on the onion bulb stem epidermis, cuts, and bulb seriously rots, and has foul smell.Test-results shows that new onion burkholderia NSM-05 is not the onion pathogenic bacterium.
Embodiment 9 new onion burkholderia NSM-05 are to the pathogenic mensuration of clover.
Alfalfa seed is soaked 20min with 98% the vitriol oil, with sterile water wash 5 times, steep then in an amount of sterilized water, 30 ℃ of concussion (120r/min) seed soaking 8h are down cleaning 2 times, under similarity condition, shake overnight cultures after changing water.Use sterile water wash 2 times next day again, aseptic technique is waited for inoculation with the alfalfa seed pendulum on 1% water agar plate, and every ware is put 10 seeds.The alfalfa seed that is placed on the water agar plate is stung an aperture with inoculating needle on cotyledon; Get the ready pure bacteria suspension of 10 μ L (new onion burkholderia NSM-05, onion burkholderia LMG1222 reference culture and sterilized water) point in the wound; Wherein onion burkholderia LMG1222 is as positive control, and sterilized water is as negative control.Each handles 3 repetitions, 10 seeds of every repetition.Postvaccinal seed is placed on the artificial climate incubator of 30 ℃, 70% relative humidity, regularly illumination (12h illumination, 12h is dark) condition, checks sickness rate behind the 8d.
The result was as shown in Figure 8 after clover inoculated 8 days, and new onion burkholderia NSM-05 and sterilized water inoculation are handled, and all do not have disease symptom, and both do not show notable difference.And onion burkholderia LMG1222 inoculation is handled, and clover shows symptoms such as necrosis, yellow and root be abnormal, and alfalfa growing obviously is suppressed.Test-results explains that new onion burkholderia NSM-05 is to the clover safety non-toxic.And clover is to replace mouse as measuring the model plant of Bcc flora to mammalian toxicity; Utilize the clover inoculation test simply rapid detection go out the Bcc bacterial strain and whether Mammals had pathogenic, above test-results explains that new onion burkholderia NSM-05 is to the mammalian safe toxicological harmless.
SEQUENCE?LISTING
< 110>Nanjing Forestry University
< 120>a kind of pine tree endogenetic bacteria new onion burkholderia NSM-05 and application thereof
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< 213>new onion burkholderia (Burkholderia cenocepacia) NSM-05
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Claims (7)
1. pine tree endogenetic bacteria, the new onion burkholderia of its classification called after (
Burkholderia Cenocepacia) NSM-05, being preserved in Chinese typical culture collection center, its deposit number is: CCTCC NO:M 2011015, preservation date: on January 11st, 2011.
2. the application of the described pine tree endogenetic bacteria of claim 1 in the biological control of pine tree shoot blight.
3. the application of the described pine tree endogenetic bacteria of claim 1 in promoting the slash pine growth.
4. sphaeropsis sapinea biocontrol fungicide; It is characterized in that this biocontrol fungicide adopts following method preparation: described new onion burkholderia NSM-05 is inoculated in substratum, air flow 1:1 with claim 1; 28 ~ 30 ℃ of temperature; Shaking table revolution 180 ~ 200r/min, 48 ~ 60h, the fermented liquid that obtains is the sphaeropsis sapinea biocontrol fungicide; Wherein, the prescription of substratum is: in every 1000ml zero(ppm) water, contain Carnis Bovis seu Bubali cream 2 ~ 5g, peptone 5 ~ 10g, sodium-chlor 5g, pH5 ~ 7.
5. the described sphaeropsis sapinea biocontrol fungicide of claim 4 is in biological control of pine tree shoot blight and the application that promotes in the slash pine growth.
6. the application of the described pine tree endogenetic bacteria of claim 1 in suppressing the forest pathogenic bacteria; Wherein, the forest pathogenic bacteria is Phomopsis and golden yellow gram capsule spore and the horse-chestnut shell shuttle spore of Ginkgo Leaf rot alternaric bacteria, Liriodendron black spot interlinkage spore, Liriodendron black spot anthrax-bacilus, dothiorella gregaria bacterium.
7. the application of the described pine tree endogenetic bacteria of claim 1 in suppressing pathogen of crop; Wherein, pathogen of crop is Rhizoctonia solani Kuhn, fusarium graminearum, cotton rhizoctonia solani, cotton-wilt fusarium, ring rot of apple bacterium, tea moire blight bacterium and tea zonate spot bacterium.
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| CN103820354B (en) * | 2013-11-21 | 2016-06-01 | 上海海洋大学 | The application of a kind of bulkholderia cepasea FH4 bacterial strain |
| CN103642742B (en) * | 2013-12-27 | 2016-01-06 | 中国热带农业科学院香料饮料研究所 | A kind of bulkholderia cepasea and application thereof |
| CN105695364B (en) * | 2016-03-28 | 2019-02-22 | 福建农林大学 | One plant of new Burkholderia cepacia and application |
| CN106495912B (en) * | 2016-11-15 | 2020-02-07 | 环境保护部南京环境科学研究所 | Method for preparing machine-transplanted rice seedling raising substrate by taking decomposed corn straws as raw materials |
| CN107099478A (en) * | 2017-06-06 | 2017-08-29 | 江苏省中国科学院植物研究所 | One plant of Lycoris aurea endogenetic bacteria bulkholderia cepasea HDXY 02 and its application |
| CN108148785B (en) * | 2018-01-31 | 2021-09-03 | 广西壮族自治区农业科学院 | Sugarcane endogenous burkholderia CZ08152 and application thereof |
| CN108739860B (en) * | 2018-05-02 | 2020-10-23 | 华南农业大学 | Microbial quorum sensing signal quenching and sterilizing agent and application thereof as biocontrol bacterium |
| CN110468084B (en) * | 2019-09-20 | 2021-04-23 | 华南农业大学 | New burkholderia cepacia and application thereof in preventing and treating litchi frost blight and litchi anthracnose |
| CN112825716B (en) * | 2019-11-22 | 2022-07-22 | 中国林业科学研究院森林生态环境与保护研究所 | Method for regulating and controlling fungal community structure for reducing harm of pine wilt disease pathogenic bacteria to host plants |
| CN112646733B (en) * | 2020-12-28 | 2023-01-17 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Tamarix chinensis endophytic antagonistic fungus as well as separation method and application thereof |
| CN112868670B (en) * | 2021-02-20 | 2021-10-22 | 南京林业大学 | Novel application of Burkholderia polyphylla WS-FJ9 |
| CN113151076B (en) * | 2021-04-13 | 2022-04-12 | 安徽省祁门县祁红茶业有限公司 | Burkholderia and application thereof |
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| CN101555458B (en) * | 2009-03-12 | 2010-09-15 | 南京林业大学 | Burkholderia pyrrocinia and application thereof in control of dothiorella gregaria |
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