CN107365723A - Chemicals and Biocontrol microorganism joint prevention and control continuous cropping tomato bacterial wilt - Google Patents
Chemicals and Biocontrol microorganism joint prevention and control continuous cropping tomato bacterial wilt Download PDFInfo
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- CN107365723A CN107365723A CN201710568541.8A CN201710568541A CN107365723A CN 107365723 A CN107365723 A CN 107365723A CN 201710568541 A CN201710568541 A CN 201710568541A CN 107365723 A CN107365723 A CN 107365723A
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- C05—FERTILISERS; MANUFACTURE THEREOF
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Abstract
The invention discloses chemicals and Biocontrol microorganism joint prevention and control continuous cropping tomato bacterial wilt, the present invention is isolated to one plant of Antagonistic Fungi (bacillus B8) for having notable antagonism to bacterial wilt of tomato pathogen, bacterial strain B8 belongs to bacillus amyloliquefaciens (Bacillus amyloliqueBaciens), China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on April 17th, 2017, culture presevation number is CGMCC NO.14042.Liquid bacterial agent and microbial organic fertilizer are produced using the Antagonistic Fungi, Antagonistic Fungi quantity is more than 10 in liquid bacterial agent9/ ml, Antagonistic Fungi quantity is more than 5.0 × 10 in microbial manure7Individual/g.Antagonistic Fungi liquid bacterial agent, the microbial organic fertilizer containing Antagonistic Fungi and the processing of 0.3% (W/V) potassium phosphite solution are used in combination in continuous cropping tomato soil, more than 70% is up to the control rate of bacterial wilt of tomato.
Description
Technical field
The present invention relates to use chemical agent, antagonism bacteria agent and microbial organic fertilizer joint prevention and control bacterial wilt of tomato, category
In the integrated control field of plant soil-borne diseases.
Background technology
Bacterial wilt of tomato is a kind of soil-borne disease as caused by blue or green withered Raul Salmonella (Ralstonia solanacearum),
The incidence of disease is generally particularly acute in 20%-30%, continuous cropping, and the incidence of disease is up to 70-80%, or even total crop failure sometimes.At present, do not have also
There is effective chemical pesticide control bacterial wilt of tomato, there is an urgent need to develop the new technology of prevention and control bacterial wilt of tomato.It is we have found that sub-
Potassium phosphate (KH2PO3), it can not only be used for phosphorus potash fertilizer, the energy grown again with the withered Raul Salmonella of suppression pathogen-green grass or young crops under finite concentration
Power, the suppression of potassium phosphite and antagonistic strain F8 to bacterial wilt have synergy, potassium phosphite and antagonistic strain F8 liquid bacterias
Agent and microbial organic fertilizer are used in combination, and can significantly improve the effect of prevention and control bacterial wilt of tomato, also comply with the need of green agriculture
Ask.
The content of the invention
It is an object of the invention to develop one kind using chemical bacteriostatic agent potassium phosphite, Antagonistic Fungi liquid bacterial agent and microorganism
The complex art of organic fertilizer prevention and control bacterial wilt of tomato, its integrated control efficiency reach 60-80%, are intensive tomato planting industry
Sustainable development provide technology.
The purpose of the present invention can be achieved through the following technical solutions：
One plant of Antagonistic Fungi B8 for being used for control of plant bacterial wilt, belongs to bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens), it is preserved in China Committee for Culture Collection of Microorganisms's commonly micro- life on April 17th, 2017
Thing center, culture presevation number are CGMCC NO.14042.Main Biological is：B8 on LB culture mediums when growing, bacterium colony
Gray, circular, bacterium colony shows smooth that there is gauffer on later stage bacterium colony surface during beginning.B8 cell is shaft-like, Later growth cell
It is interior to form spherical gemma.Antagonistic Fungi B8 bacterial strains have glucose fermentation, fructose, maltose, the ability of sucrose production acid, can hydrolyze
Starch and liquefaction gelatin.
Applications of the above-mentioned Antagonistic Fungi B8 in the biological prevention and control agent for prevention and control bacterial wilt of tomato is prepared.
The biological prevention and control agent for prevention and control bacterial wilt of tomato produced using above-mentioned Antagonistic Fungi B8.
Applications of the above-mentioned Antagonistic Fungi B8 in prevention and control bacterial wilt of tomato.
Applications of the above-mentioned Antagonistic Fungi B8 in chemicals potassium phosphite prevention and control bacterial wilt of tomato is combined.
Above-mentioned application, comprises the following steps：(1) B8 liquid bacterial agents are prepared using above-mentioned Antagonistic Fungi B8 respectively and B8 is micro-
Biological organic fertilizer；(2) the B8 liquid bacterial agents and concentration prepared step (1) is 10%~30% (W/V, 10~30g/100ml)
Starch solution according to 1:1 volume ratio is mixed and made into mixed liquor；(3) the mixed liquor root dipping for preparing tomato seedling with step (2),
Make Antagonistic Fungi B8 cell adherences in tomato seedling root；(4) when root is stained with the tomato transplantation of seedlings of Antagonistic Fungi B8 cells, by step
(1) the B8 microbial organic fertilizers prepared are applied in tomato seedling root, and concentration is finally poured in the root of every plant of tomato seedling as 0.2%
~0.4% (W/V, 0.2~0.4g/100ml) potassium phosphite solution.
A kind of method of efficiently prevention and control continuous cropping tomato bacterial wilt, comprises the following steps：
(1) B8 liquid bacterial agents and B8 microbial organic fertilizers are prepared using above-mentioned Antagonistic Fungi B8 respectively；
(2) the B8 liquid bacterial agents and concentration that prepare step (1) are 10%~30% (W/V, 10~30g/100ml) shallow lake
Powder solution is according to 1:1 volume ratio is mixed and made into mixed liquor；
(3) the mixed liquor root dipping for preparing tomato seedling with step (2), Antagonistic Fungi B8 cell adherences are made in tomato seedling root；
(4) when root is stained with the tomato transplantation of seedlings of Antagonistic Fungi B8 cells, by the B8 microbial organic fertilizers of step (1) preparation
Tomato seedling root is applied in, it is 0.2%~0.4% (W/V, 0.2~0.4g/ that concentration is finally poured in the root of every plant of tomato seedling
Potassium phosphite solution 100ml).
Above-mentioned method, the concentration of starch solution described in its step (2) are 20%；Potassium phosphite described in step (4)
The concentration of solution is 0.3%.
Above-mentioned method, the amount of application of B8 microbial organic fertilizers described in its step (4) are 40~60g/；The phosphorous
The amount of pouring of sour potassium solution is 40~60ml/.
Above-mentioned method, the middle B8 liquid bacterial agents miospore and cell prepared using above-mentioned Antagonistic Fungi B8 of its step (1)
Quantity reach 109/ ml the orders of magnitude；Contain 5 in the B8 microbial organic fertilizers prepared in step (1) using above-mentioned Antagonistic Fungi B8
×107Individual/more than g Antagonistic Fungi F8 spore.
With the specific life of antagonistic bacterial strains B8 production B8 liquid bacterial agents and B8 microbial organic fertilizers in step (1) of the present invention
Production method may refer to patent ZL2012 1 0326641.7, but not limited to this, can also use as well known to those skilled in the art
Other method production.
Potassium phosphite used is technical grade in the above method, and concentration is 0.2%~0.4%, preferably 0.3%
(W/V).Potassium phosphite (the KH of present invention experiment display 0.3%2PO3) significantly inhibit the growth of the withered Raul Salmonella of pathogen-green grass or young crops
(see Fig. 2), and biocontrol bacterial strain B8 is not influenceed.
The most preferred method being used in combination of biocontrol bacterial strain B8 liquid bacterial agent, microbial organic fertilizer and potassium phosphite：
By B8 liquid bacterial agents and 20% (W/V) starch solution according to 1:1 (V/V) ratio mixing, for tomato seedling root dipping, makes B8 thin
Born of the same parents are adhered to tomato seedling root.B8 microbial organic fertilizers are using spreading manuer in holes, according to every seedling 50g usage amount by microbial organic fertilizer
Tomato seedling root, then earthing are sprinkling upon, 0.3% (W/V) potassium phosphite solution 50ml is finally poured every plant of tomato seedling root.
Beneficial effects of the present invention：
The antagonistic bacterial strains B8 that the present invention screens has significant control effect to bacterial wilt of tomato, using bacterial strain B8 systems
Biological control of the standby biological prevention and control agent to bacterial wilt of tomato is notable, and the result of pot experiment and field test shows, continuous cropping tomato
After synthesis applies B8 liquid bacterial agents, B8 microbial organic fertilizers and potassium phosphite in the soil of bacterial wilt, the life of bacterial wilt of tomato
Thing control rate is up to more than 70%, shows that Combined Treatment has significant synergistic function, is remarkably improved withered to tomato green grass or young crops
The prevention effect of disease.
Brief description of the drawings
Fig. 1 is that (display presses down inhibitory action of the antagonistic bacterial strains B8 to bacterial wilt of tomato pathogen (R.solanacearum)
Bacterium circle)
Fig. 2 is that (display presses down inhibitory action of 0.3% potassium phosphite to bacterial wilt of tomato pathogen (R.solanacearum)
Bacterium circle)
Fig. 3 is effect of the Combined Treatment to control of plant bacterial wilt
Embodiment
(1) separation and identification of bacterial strain
Plate dilution method separation of bacterial is used from tomato plant rhizosphere soil, then with bacterial wilt of tomato pathogen-green grass or young crops
Withered Raul Salmonella (Ralstonia solanacearum) is that indicator bacteria screens Antagonistic Actinomycetes using face-off method, according to growth speed
Degree, antagonistic ability finally filter out efficient antagonistic strain B8, by morphological feature, physio-biochemical characteristics and 16S rRNA genes
Bacterial strain B8 is accredited as bacillus amyloliquefaciens by sequence.
Bacterial strain B8 was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on April 17th, 2017
The heart, culture presevation number are CGMCC NO.14042.Main Biological is：For F8 when being grown on LB culture mediums, bacterium colony is in ash
Color, circular, bacterium colony shows smooth that there is gauffer on later stage bacterium colony surface during beginning.B8 cell is shaft-like, the intracellular shape of Later growth
The gemma of globulate.Antagonistic Fungi B8 bacterial strains have glucose fermentation, fructose, maltose, the ability of sucrose production acid, can hydrolysis starch
With liquefaction gelatin.
Antagonistic bacterial strains B8 is as shown in Figure 1 to the inhibitory action of bacterial wilt of tomato pathogen (R.solanacearum).
(2) production of bacterial strain B8 liquid bacterial agents and microbial organic fertilizer
Production method is referring to patent ZL2012 1 0326641.7.
1) preparation of B8 liquid bacterial agents：Bacterial strain B8 is inoculated into progress liquid fermentation production in PDA nutrient solutions, its life of fermenting
The condition of production is：Initial pH scopes be 7.0-7.2,30 DEG C of cultivation temperature, 170 revs/min, form gemma after the 48h that ferments, acquisition
B8 liquid bacterial agents, it is therein to contain bacterium or gemma amount >=5 × 109Individual/ml；
PDA cultures liquid making method used is, exemplified by preparing 1L culture mediums：It is cut into small pieces after being peeled with 200g potatoes and puts
Boiled in water, 30min is boiled after boiling, added 20g common sucroses in filtrate after filtering, be settled to 1000ml, pH value is adjusted to 7.2-
7.4,121 DEG C of sterilizing 20min.
2) preparation of B8 microbial organic fertilizers：With decomposed pig manure and amino acid fertilizer (Jiangsu new world amino
Sour fertilizer Co., Ltd product, registration card number：The quasi- words of Su Nongfei (2008) 0180, fertilizer specification：N+P2O5+K2O >=6%, have
Machine matter >=30%, amino acid >=5%) with volume ratio 6：4 are mixed.Decomposed pig manure germination index used be 98% with
On, quality of organic matter is 1.2-2%, water content 25-30% than content than content >=35%, organic nitrogen quality.
B8 liquid bacterial agents prepared by step 1) are inoculated into described in step 2) by 20% (V/W, 20ml/100g) ratio
Secondary solid fermentation is carried out in organic fertilizer, fermentation temperature is 30-50 DEG C, and turning daily 1 time in fermentation process, ferment fermentation period
5-7 days, antagonism bacterial content is set to reach 2.0 × 109Individual/more than g, obtain Antagonistic Fungi B8 solid fungicide.Do not surpass in temperature finally
Cross under conditions of 60 DEG C and the water content of microbial organic fertilizer is evaporated to less than 30%, obtain the micro- lifes of F8 of control of plant bacterial wilt
Organic fertilizer, >=2.0 × 10 are contained in the fertilizer9Individual/g Antagonistic Fungi B8, total nitrogen content 30.9g.kg-1, content of tatal phosphorus is
12.0g.kg-1, full potassium content be 20.9g.kg-1。
(3) greenhouse pot culture is tested
It is " Hezuo 918 " for examination tomato variety, kind of a tomato species industry Co., Ltd is grown by Shanghai and produced.4~5 true leaves of nursery
It is transplanted to afterwards in the basin alms bowl equipped with 5kg soil.4 processing are set：
1)CK；
2) potassium phosphite of B8 liquid bacterial agents+B8 microbial organic fertilizers+0.3% is handled
3) B8 liquid bacterial agents+F8 microbial organic fertilizers：By B8 liquid bacterial agents and 20% (W/V) starch solution according to 1:1
(V/V) ratio mixing, for tomato seedling root dipping, makes bacterial strain B8 cell adherences in tomato seedling root.Microbial organic fertilizer uses
Spread manuer in holes, microbial organic fertilizer is sprinkling upon tomato seedling root, then earthing according to every seedling 50g usage amount.
4) 0.3% potassium phosphite is handled：CK processing nitrogen, phosphorus, potash fertilizer polishing nutrient, are poured every plant of tomato seedling root
0.3% (W/V) potassium phosphite solution 50ml.
Every kind of processing sets three district's groups, each 15 repetitions of district's groups.Pathogen is inoculated with soil, content is 8.5 × 106
Individual/g soil.The process of the potassium phosphite integrated treatment of B8 liquid bacterial agent+B8 microbial organic fertilizers+0.3% is：By B8 liquid bacterial agents and
20% (W/V) starch solution is according to 1:1 (V/V) ratio mixing, for tomato seedling root dipping, make bacterial strain B8 cell adherences kind
Eggplant seedling root.Microbial organic fertilizer is sprinkling upon tomato seedling root by microbial organic fertilizer using spreading manuer in holes according to every seedling 50g usage amount
Portion, then earthing, 0.3% (W/V) potassium phosphite solution 50ml (0.3% potassium phosphites pair are finally poured every plant of tomato seedling root
The inhibitory action of bacterial wilt of tomato pathogen (R.solanacearum) is shown in Fig. 2).CK processing nitrogen, phosphorus, potash fertilizer polishing nutrient.
A situation arises for investigation bacterial wilt daily after 20 days for tomato growth, calculates disease index and prevention effect：
Disease index=∑ (diseased plant numbers at different levels × corresponding series)/total plant number × 100 of investigation；
Prevention effect=(check plot disease index-treatment region disease index)/check plot disease index × 100；
Disease severity grade scale：0 grade：Without symptom；I grade：1/4 blade is fallen ill；II grade：1/2 blade is fallen ill；III grade：
Whole strain is dead.
As a result show, it is 40.6% to the prevention effect of bacterial wilt of tomato that 0.3% potassium phosphite, which is used alone, uses B8 liquid
Body microbial inoculum and B8 microbial organic fertilizers are 51.1% to the prevention effect of bacterial wilt of tomato, and B8 liquid bacterial agents, B8 is administered in combination
The integrated treatment of microbial organic fertilizer and potassium phosphite reaches 76.2% (table 1) to the prevention effect of bacterial wilt of tomato, shows to join
Closing processing has synergy, is remarkably improved the prevention effect (Fig. 3) to bacterial wilt of tomato.
Effect of the Combined Treatment of table 1 to control of plant bacterial wilt
| Processing | Disease index (%) | Prevention effect (%) |
| CK | 90.6 | — |
| Integrated treatment | 21.5 | 76.2 |
| Comparative example 1 (B8 liquid bacterial agent+B8 microbial organic fertilizers) | 44.3 | 51.1 |
| Comparative example 2 (processing of 0.3% potassium phosphite) | 53.8 | 40.6 |
Claims (10)
1. one plant of Antagonistic Fungi B8 for being used for prevention and control bacterial wilt of tomato, bacterial strain B8 belong to bacillus amyloliquefaciens (Bacillus
AmyloliqueBaciens), it is preserved in China Committee for Culture Collection of Microorganisms's commonly micro- life on April 17th, 2017
Thing center, culture presevation number are CGMCC NO.14042.
2. applications of the Antagonistic Fungi B8 in the biological prevention and control agent for prevention and control bacterial wilt of tomato is prepared described in claim 1.
3. using the biological prevention and control agent for prevention and control bacterial wilt of tomato of the Antagonistic Fungi B8 productions described in claim 1.
4. applications of the Antagonistic Fungi B8 in prevention and control bacterial wilt of tomato described in claim 1.
5. applications of the Antagonistic Fungi B8 in chemicals potassium phosphite prevention and control bacterial wilt of tomato is combined described in claim 1.
6. application according to claim 5, it is characterised in that：Comprise the following steps：
(1) B8 liquid bacterial agents and B8 microbial organic fertilizers are prepared using the Antagonistic Fungi B8 described in claim 1 respectively；
(2) starch solution that the B8 liquid bacterial agents and concentration prepared step (1) is 10%~30% is according to 1:1 volume ratio is mixed
Mixed liquor is made in conjunction；
(3) the mixed liquor root dipping for preparing tomato seedling with step (2), Antagonistic Fungi B8 cell adherences are made in tomato seedling root；
(4) when root is stained with the tomato transplantation of seedlings of Antagonistic Fungi B8 cells, B8 microbial organic fertilizers prepared by step (1) are applied
In tomato seedling root, the potassium phosphite solution that concentration is 0.2%~0.4% is finally poured in the root of every plant of tomato seedling.
A kind of 7. method of efficiently prevention and control continuous cropping tomato bacterial wilt, it is characterised in that：Comprise the following steps：
(1) B8 liquid bacterial agents and B8 microbial organic fertilizers are prepared using the Antagonistic Fungi B8 described in claim 1 respectively；
(2) starch solution that the B8 liquid bacterial agents and concentration prepared step (1) is 10%~30% is according to 1:1 volume ratio is mixed
Mixed liquor is made in conjunction；
(3) the mixed liquor root dipping for preparing tomato seedling with step (2), Antagonistic Fungi B8 cell adherences are made in tomato seedling root；
(4) when root is stained with the tomato transplantation of seedlings of Antagonistic Fungi B8 cells, B8 microbial organic fertilizers prepared by step (1) are applied
In tomato seedling root, the potassium phosphite solution that concentration is 0.2%~0.4% is finally poured in the root of every plant of tomato seedling.
8. according to the method for claim 7, it is characterised in that：The concentration of starch solution described in step (2) is 20%；Step
Suddenly the concentration of potassium phosphite solution described in (4) is 0.3%.
9. the method according to claim 7 or 8, it is characterised in that：The administration of B8 microbial organic fertilizers described in step (4)
Measure as 40~60g/；The amount of pouring of the potassium phosphite solution is 40~60ml/.
10. according to the method for claim 7, it is characterised in that：Using the Antagonistic Fungi described in claim 1 in step (1)
B8 liquid bacterial agents miospore and the quantity of cell prepared by B8 reaches 109/ ml the orders of magnitude；Using claim 1 institute in step (1)
Contain 5 × 10 in B8 microbial organic fertilizers prepared by the Antagonistic Fungi B8 stated7Individual/more than g Antagonistic Fungi B8 spore.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019184259A1 (en) * | 2018-03-25 | 2019-10-03 | 江苏辉丰生物农业股份有限公司 | Medical fertilizer composition containing potassium phosphite and polyglutamic acid |
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| WO2019184259A1 (en) * | 2018-03-25 | 2019-10-03 | 江苏辉丰生物农业股份有限公司 | Medical fertilizer composition containing potassium phosphite and polyglutamic acid |
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