阅读说明：本技术 一种植物用复合菌剂及其制备方法 (A kind of plant composite bacteria agent and preparation method thereof ) 是由 束震 吕超田 束张静怡 束冰 于 2019-08-14 设计创作，主要内容包括：本发明公开了一种植物用复合菌剂及其制备方法,涉及植物复合菌剂技术领域,本发明以多黏芽孢杆菌微囊粒剂和木霉菌微囊粒剂复配剂对田间已发生枯萎病的西瓜苗进行灌根防治,经芽孢杆菌、木霉菌复配处理后与不使用任何药剂的对照相比西瓜枯萎病发病率明显降低,二者的复配药剂均可显著促进西瓜的生长发育,培育健壮植株,同时采用微囊粒剂具有货架期长,药效稳定,缓慢释放有效成分分解和挥发,减少用药量,防止漂移,减少环境污染,使用方便,提高药剂选择性等作用,是最具发展前景的剂型之一。(The invention discloses a kind of plant composite bacteria agents and preparation method thereof, it is related to plant composite bacteria agent technical field, the present invention carries out pouring root prevention and treatment with the watermelon seedling that field wilt disease has occurred for Bacillus polymyxa microcapsulated granule and Trichoderma microcapsulated granule compounding agent, through bacillus, watermelon blight disease incidence is substantially reduced compared with the control without using any medicament after Trichoderma complex treatment, the compound drug of the two can significantly promote the growth and development of watermelon, cultivate healthy and strong plant, have shelf life long using microcapsulated granule simultaneously, efficacy stability, slow release effective component is decomposed and volatilization, reduce dosage, it prevents from drifting about, it reduces environmental pollution, it is easy to use, improve the effects of medicament is selective, it is one of dosage form most with prospects.)

1. a kind of plant composite bacteria agent, which is characterized in that including following mass percent ingredient, mostly glutinous bacillus micro-capsule grain Agent 50~75%, Trichoderma microcapsulated granule 25~50%.

2. plant composite bacteria agent according to claim 1, which is characterized in that following mass percent ingredient is included, it is mostly glutinous Bacillus microcapsulated granule 60%, Trichoderma microcapsulated granule 40%.

3. plant composite bacteria agent according to claim 1, which is characterized in that more glutinous bacillus microcapsulated granules and The living bacteria count of Trichoderma microcapsulated granule is not less than 1,000,000,000 CFU/g.

4. plant composite bacteria agent according to claim 1, which is characterized in that the Trichoderma is May 23 in 2019 Day is deposited in China typical culture collection center, and deposit number is CCTCC NO:M2019337.

5. a kind of preparation method of plant composite bacteria agent, which is characterized in that be prepared using the method included the following steps:

Microcapsulated granule will stick bacillus and Trichoderma more and successively expand culture and the culture of second level tank by first order seed respectively Afterwards, starch and diatomite is then added into the fermentor of the two respectively, gelatinization mixing is stirred at 20~30 DEG C of temperature After uniformly, two kinds of dextrins are then passed through into the microcapsulated granule that cryogenic vacuum air prilling technology is made smaller than 200 μm respectively, Obtain more glutinous bacillus microcapsulated granules and Trichoderma microcapsulated granule；

The how glutinous bacillus microcapsulated granule and Trichoderma microcapsulated granule of preparation are added to by corn pulp by the preparation of composite bacteria agent In the solution that the temperature that dry powder, glucose, ammonium sulfate and potassium humate form is 8~15 DEG C, it is uniformly mixed, while using phosphorus The pH that acid dihydride potassium adjusts mixed liquor is not less than 5, obtains composite bacteria agent.

6. plant according to claim 5 composite bacteria agent preparation method, which is characterized in that the starch and diatomite Mass ratio is 5~15:1, and the aperture of diatomite is greater than the partial size of starch.

7. plant according to claim 5 composite bacteria agent preparation method, which is characterized in that the Dried Corn Steep Liquor Powder, Portugal The mass ratio of grape sugar, ammonium sulfate and potassium humate is 5:1:1:3.

8. plant according to claim 5 composite bacteria agent preparation method, which is characterized in that the composite bacteria agent it is effective Viable count is not less than 1,000,000,000 CFU/g.

9. a kind of application of plant composite bacteria agent, which is characterized in that for preventing and treating watermelon blight and promoting watermelon growing.

Technical field

The present invention relates to plant composite bacteria agent technical field, a kind of plant composite bacteria agent and its preparation side are particularly related to Method.

Background technique

Watermelon blight claims that dead seedling disease, wheat cuts disease, stem wilt, is by watermelon specialized form Fusarium oxysporum (Fusarium Oxysporum f.sp.niveum) infect caused soil-borne disease, cause of disease can time-to-live 8~10 in the soil Year.Watermelon entire breeding time all can be caused the yield of watermelon to decline 30%, some plot underproduction by the harm of wilt disease 50% or more, or even total crop failure.Seriously affect high-quality, the efficient production of watermelon.Moreover, with the expansion of growth of watermelon area, west Melon continuous cropping continuous cropping has also been continuously increased the generation of watermelon blight.Do not have for the prevention and treatment of watermelon blight both at home and abroad at present The method of effect is the cancer in watermelon production.It is also inadequate using the methods of grafting and root-changing, breeding for disease resistance, crop rotation, soil removal and replacement effect Ideal, it is at high cost, demand strict technology.Agro-chemicals control effect of chemical control is poor.And biological control is excellent in addition to having Except preventive effect, also have protect and improve farmland ecological environment, it is free from environmental pollution, it is safe to people, animal the advantages that.

With the continuous promotion of the development degree of agricultural, microbial pesticide dosage form (based on composite bacteria agent) is also constantly gushed Reveal and, but applicant has found in research, it is domestic at present not deep enough to the research of the related microorganisms formulations of pesticide, mainly It is single to show as dosage form, lacks high-caliber dosage form, restricts a most important factor for microbial pesticide dosage form development at present It is exactly that there are shelf life is short for microbial pesticide dosage form, is easy to happen medicinal powder when in use and flies upward, cause damages to personnel.

Summary of the invention

In view of this, existing to solve it is an object of the invention to propose a kind of plant composite bacteria agent and preparation method thereof Have in technology that microbial pesticide dosage form shelf life is short, the used time is easy to happen the problems such as medicinal powder is flown upward, and withered in prevention and treatment watermelon Application in disease.

It is more based on a kind of above-mentioned purpose plant composite bacteria agent provided by the invention, including following mass percent ingredient Glutinous bacillus microcapsulated granule 50~75%, Trichoderma microcapsulated granule 25~50%.

Optionally, following mass percent ingredient is included, mostly glutinous bacillus microcapsulated granule 60%, Trichoderma microcapsulated granule 40%.

Optionally, the living bacteria count of more glutinous bacillus microcapsulated granules and Trichoderma microcapsulated granule is not less than 10 Hundred million CFU/g.

Optionally, the Trichoderma is deposited in China typical culture collection center, preservation on May 23rd, 2019 It number is CCTCC NO:M2019337.

A kind of preparation method of plant composite bacteria agent, is prepared using the method included the following steps:

Microcapsulated granule will stick bacillus and Trichoderma more and successively expand culture and second level tank by first order seed respectively After culture, starch and diatomite is then added into the fermentor of the two respectively, is stirred gelatinization at 20~30 DEG C of temperature After mixing, two kinds of dextrins are then passed through into the micro-capsule that cryogenic vacuum air prilling technology is made smaller than 200 μm respectively Granula obtains more glutinous bacillus microcapsulated granules and Trichoderma microcapsulated granule；

The preparation of composite bacteria agent, by the how glutinous bacillus microcapsulated granule and Trichoderma microcapsulated granule of preparation be added to by In the solution that the temperature that Dried Corn Steep Liquor Powder, glucose, ammonium sulfate and potassium humate form is 8~15 DEG C, it is uniformly mixed, It is not less than 5 with the pH that potassium dihydrogen phosphate adjusts mixed liquor simultaneously, obtains composite bacteria agent.

Optionally, the mass ratio of the starch and diatomite is 5~15:1, and the aperture of diatomite is greater than the grain of starch Diameter.

Optionally, the mass ratio of the Dried Corn Steep Liquor Powder, glucose, ammonium sulfate and potassium humate is 5:1:1:3.

Optionally, the living bacteria count of the composite bacteria agent is not less than 1,000,000,000 CFU/g.

A kind of application of plant composite bacteria agent, for preventing and treating watermelon blight and promoting watermelon growing.

From the above it can be seen that a kind of plant composite bacteria agent provided by the invention and preparation method thereof, mostly glutinous bud Spore bacillus can secrete generation extracellular antiseptic protein, have suppression to various crop Sclerotium rolfsii spore germinations and mycelia growth Production is used, and Trichoderma is also the generally existing biocontrol fungi of nature, can pass through hyperparasitism, the mechanism such as antibiosis Play the role of inhibiting and killing to germ；Simultaneously Bacillus polymyxa and Trichoderma all in edaphon beneficial to micro- life There is certain mutual synergistic effect between object, possible disease resistance mechanisms are and Fusarium oxysporum is in living space and nutrition side There are certain Competitions in face；So as to fight for and plant with pathogen in the internal of plant, rhizosphere and body surface mass propagation Nutrition around object prevents pathogen from infecting plant organ, and then inhibits the reproduction speed of pathogen；Also can simultaneously Disease resistance mechanisms existing for enough indirect induction plants itself reduce plant diseased plant number, and this hair to resist the attack of pathogen Bright to have shelf life long using microcapsulated granule, efficacy stability, slow release effective component is decomposed and volatilization, reduces dosage, prevents It only drifts about, reduces environmental pollution, it is easy to use, the effects of medicament is selective is improved, is one of dosage form most with prospects.

Specific embodiment

For below by the description to embodiment, for example related manufacturing process of a specific embodiment of the invention and behaviour Make application method etc., be described in further detail, to help those skilled in the art to inventive concept of the invention, technology Scheme has more complete, accurate and deep understanding.

Need to illustrate: Trichoderma used in this application is preserved in China typical culture collection center, address: Chinese Wuhan Wuhan University；The deposit date is on May 23rd, 2019, deposit numbers: CCTCC NO.M2019337, preservation biological classification Title: Trichoderma harzianum (Trichoderma harzianum).

In order to solve all or part of deficiency of composite bacteria in the prior art, a kind of plant use provided by the invention Composite bacteria agent, including following mass percent ingredient, mostly glutinous bacillus microcapsulated granule 50~75%, Trichoderma microcapsulated granule 25~50%.

The preparation method of a kind of plant composite bacteria agent provided simultaneously, using the method preparation included the following steps At:

Microcapsulated granule will stick bacillus and Trichoderma more and successively expand culture and second level tank by first order seed respectively After culture, starch and diatomite is then added into the fermentor of the two respectively, is stirred gelatinization at 20~30 DEG C of temperature After mixing, two kinds of dextrins are then passed through into the micro-capsule that cryogenic vacuum air prilling technology is made smaller than 200 μm respectively Granula obtains more glutinous bacillus microcapsulated granules and Trichoderma microcapsulated granule；

The preparation of composite bacteria agent, by the how glutinous bacillus microcapsulated granule and Trichoderma microcapsulated granule of preparation be added to by In the solution that the temperature that Dried Corn Steep Liquor Powder, glucose, ammonium sulfate and potassium humate form is 8~15 DEG C, it is uniformly mixed, It is not less than 5 with the pH that potassium dihydrogen phosphate adjusts mixed liquor simultaneously, obtains composite bacteria agent.

Composite bacteria is for preventing and treating watermelon blight and promoting watermelon growing.

Bacillus polymyxa can secrete generation extracellular antiseptic protein, to various crop Sclerotium rolfsii spore germinations and Mycelia growth has inhibiting effect, and Trichoderma is also the generally existing biocontrol fungi of nature, can by hyperparasitism, The mechanism such as antibiosis play the role of inhibiting and killing to germ；Simultaneously Bacillus polymyxa and Trichoderma all with the micro- life of soil There is certain mutual synergistic effect between beneficial microbe in object, possible disease resistance mechanisms are and Fusarium oxysporum is in life Deposit in terms of space and nutrition that there are certain Competitions；So as in the internal of plant, rhizosphere and body surface mass propagation, Nutrition with around pathogen contention plant, prevents pathogen from infecting plant organ, and then inhibit the breeding of pathogen Speed；Simultaneously also can disease resistance mechanisms existing for indirect induction plant itself to resist the attack of pathogen reduce plant Diseased plant number, and the present invention has shelf life long using microcapsulated granule, efficacy stability, slow release effective component is decomposed and volatilization, Dosage is reduced, prevents from drifting about, reduce environmental pollution, it is easy to use, the effects of medicament is selective is improved, is before most developing One of dosage form of scape.

The further embodiment of the present invention 1 provides a kind of plant composite bacteria agent, including following mass percent ingredient, Mostly glutinous bacillus microcapsulated granule 60%, Trichoderma microcapsulated granule 40%, and mostly glutinous bacillus microcapsulated granule and Trichoderma are micro- 1,000,000,000 CFU/g of living bacteria count of capsule granula.

A kind of preparation method of plant composite bacteria agent, is prepared using the method included the following steps:

Microcapsulated granule will stick bacillus and Trichoderma more and successively expand culture and second level tank by first order seed respectively After culture, starch and diatomite is then added into the fermentor of the two respectively, is stirred gelatinization at 20~30 DEG C of temperature After mixing, two kinds of dextrins are then passed through into the micro-capsule that cryogenic vacuum air prilling technology is made smaller than 200 μm respectively Granula obtains more glutinous bacillus microcapsulated granules and Trichoderma microcapsulated granule；

The preparation of composite bacteria agent, by the how glutinous bacillus microcapsulated granule and Trichoderma microcapsulated granule of preparation be added to by In 10 DEG C of temperature of the solution that Dried Corn Steep Liquor Powder, glucose, ammonium sulfate and potassium humate form, it is uniformly mixed, uses simultaneously The pH that potassium dihydrogen phosphate adjusts mixed liquor is not less than 5, obtains composite bacteria agent.

The mass ratio of starch and diatomite is 10:1, and the aperture of diatomite is greater than the partial size of starch；Dried Corn Steep Liquor Powder, The mass ratio of glucose, ammonium sulfate and potassium humate is 5:1:1:3；The living bacteria count of composite bacteria agent is not less than 1,000,000,000 CFU/ g。

Optionally, the embodiment of the invention also provides a kind of plant composite bacteria agent, including following mass percent at Point, mostly glutinous bacillus microcapsulated granule 50%, Trichoderma microcapsulated granule 50%, while using the method for following performance test It is tested for the property, the result of test is that main stem rugosity can increase by 35.68%, blade face transverse diameter, vertical diameter increases separately 14.55% With 14.82%, fruit-setting rate also increases by 35.8%, preventive effect 82.12%.

Optionally, the embodiment of the invention also provides a kind of plant composite bacteria agent, including following mass percent at Point, mostly glutinous bacillus microcapsulated granule 75%, Trichoderma microcapsulated granule 25%.Simultaneously using the method for following performance test It is tested for the property, the result of test is that main stem rugosity can increase by 41.68%, blade face transverse diameter, vertical diameter increases separately 14.75% With 14.82%, fruit-setting rate also increases by 38.5%, preventive effect 82.53%.

Optionally, the embodiment of the invention also provides a kind of plant composite bacteria agent, including following mass percent at Point, mostly glutinous bacillus microcapsulated granule 70%, Trichoderma microcapsulated granule 30%.Simultaneously using the method for following performance test It is tested for the property, the result of test is that main stem rugosity can increase by 38.68%, blade face transverse diameter, vertical diameter increases separately 14.45% With 14.52%, fruit-setting rate also increases by 37.4%, preventive effect 82.45%.

Optionally, the embodiment of the invention also provides a kind of plant composite bacteria agent, including following mass percent at Point, mostly glutinous bacillus microcapsulated granule 55%, Trichoderma microcapsulated granule 45%.Simultaneously using the method for following performance test It is tested for the property, the result of test is that main stem rugosity can increase by 38.14%, blade face transverse diameter, vertical diameter increases separately 13.58% With 13.75%, fruit-setting rate also increases by 36.45%, preventive effect 82.13%.

Performance test

1, test material, test plant are watermelon, and reagent agent is the how glutinous bacillus microcapsulated granule of 1,000,000,000 CFU/g The composite bacteria agent of the 1000000000 CFU/g compounding prepared with the Trichoderma microcapsulated granule and embodiment 1 of 1,000,000,000 CFU/g, mostly glutinous bud Born of the same parents bacillus and Trichoderma are screened by applicant, separated and identified.Control chemical agent is Fluoxastrobin.

2, test method

Testing has watermelon blight disease symptom in Anhui Province, Fengyang County, Bengbu on May 18th, 2019 to June 14 Field soil in carry out.Test sets 5 groups: (1) not using the soil (CK) of any medicament, the mostly glutinous gemma of (2) 10 hundred million CFU/g 300 times of liquid of bacillus microcapsulated granule, (3) 10 hundred million 300 times of liquid of CFU/g Trichoderma microcapsulated granule, (4) embodiment 1 prepare 1,000,000,000 300 times of liquid of composite bacteria agent of CFU/g compounding, random alignment, 2 repetitions of each processing.Each treatment region application is every using measuring cup Strain pouring root 500mL carries out second by same procedure and dose from after the 7d of application for the first time and is administered, and (5) use at Fluoxastrobin Reason trial zone periphery watermelon seedling soil does chemical agent control, other Routine Managements.

3, preventive effect measures

It is observed and recorded everywhere when 20d (June 14) before being administered for the first time after (May 18) and second application The incidence of area's watermelon is managed, and (Wang Hanrong, Fang Li watermelon and stock variety (are) identification of anti-blight referring to Wang Hanrong With the north evaluation [J] gardening, 2010, (8)；181-183) method, disease scale standard: 0 grade: without illness；1 grade: 1 or 2 Piece cotyledon slightly turns to be yellow, but grows normal；There is necrotic plaque in 2 grades: 1~2 yellowing leafs or 1 cotyledon, and true leaf slightly turns yellow, There is slight wilt；3 grades: cotyledon is all withered, and true leaf is obviously wilted, growth retardation；4 grades: complete stool is seriously wilted or death. Disease index, disease incidence, control efficiency are calculated according to investigation result and according to following formula.

Disease incidence=diseased plant number/investigation total strain number × 100%

Disease index=[∑ (diseased plant numbers at different levels × corresponding series)/(investigating total plant number × highest series)] × 100%

Control efficiency=[(check plot disease index-treatment region disease index)/check plot disease index] × 100%

4, the measurement of the main growth indexes of watermelon

Each treatment region watermelon plant main stem length is measured after the test, main stem diameter, the vertical diameter in blade face, blade face transverse diameter, is sat Fruit number.Every processing randomly selects 3 plants, indulges diameter, blade face transverse diameter using tape measure measurement main stem length, blade face, is surveyed using vernier caliper Determine main stem diameter.Main stem length is length of the cotyledon to main stem top.Main stem diameter intermediate position between main stem protelum is straight Diameter.

Test result

1, to the control efficiency of watermelon blight

Preventive effect of 1 different agents of table to watermelon blight

The result shows that using the compound drug of 300 times of liquid of Bacillus polymyxa, 300 times of liquid of Trichoderma and two kinds of preparations After 500mL is to watermelon pouring root 20d, watermelon plant wilt disease disease incidence before being administered than reducing, and each processing disease incidence Significantly lower than the control efficiency of blank control.The compounding of Bacillus polymyxa, Trichoderma and Bacillus polymyxa and Trichoderma 300 times of liquid disease indexs of test medicine are respectively 10.78,9.62,6.14.The disease index of blank control treatment region is 16.49, the disease index of three kinds of microbial inoculums processing is significantly lower than control treatment group.The preventive effect highest of composite bacteria agent processing, reaches 82.78%, have good preventive and therapeutic effect, especially therapeutic effect significant.

2, to the growth-promoting functions of watermelon

Influence of 2 different agents of table to watermelon growing

Influence investigation discovery (table 2) of five processing to the growing way of watermelon, Bacillus polymyxa, Trichoderma and two kinds The compounding post-processing of preparation water melon leaf is dark green, robust plant, number of bearing fruit are more, and vines and leaf growth are very fast, integrate institute There are a data, composite bacteria agent treated main stem length, main stem rugosity, blade face transverse diameter, diameter is indulged on blade face and fruit-setting rate is significantly higher than Blank control and chemical agent.Blade face longitudinal and transverse demeter increases by 15% or more than blank control.Bacillus polymyxa and Trichoderma are single The growing way and no significant difference of watermelon between the two after reason of staying alone, but main stem is sturdy, and blade transverse diameter and vertical diameter are also all larger than blank pair According to.Inhibiting effect is played to watermelon main stem length after the processing of chemical pesticide Fluoxastrobin.It is compound from the point of view of the number of bearing fruit of investigation Microbial inoculum is beared fruit several highests, all in all plays apparent facilitation to the growing way of watermelon after three microbial inoculums processing.Wherein Composite bacteria agent processing is had outstanding performance, and main stem rugosity can increase by 42.68%, blade face transverse diameter, vertical diameter increases separately 15.75% and 15.82%, fruit-setting rate also increases by 41.5%.

Detection discovery is carried out to the 20d after the 2nd application simultaneously, applies Bacillus polymyxa microcapsulated granule and trichoderma Phytotoxicity does not occur for the treatment region of bacterium microcapsulated granule and composite bacteria agent, show the drug formulation of the embodiment of the present invention, method and Dosage is to watermelon growing safety.

It should be understood by those ordinary skilled in the art that: the discussion of any of the above embodiment is exemplary only, not It is intended to imply that the scope of the present disclosure (including claim) is limited to these examples；Under thinking of the invention, above embodiments Or can also be combined between the technical characteristic in different embodiments, step can be realized with random order, and be existed such as Many other variations of the upper different aspect of the invention, for simplicity, they are not provided in details.

What the embodiment of the present invention was intended to cover fall within the broad range of appended claims all such replaces It changes, modifications and variations.Therefore, all within the spirits and principles of the present invention, any omission for being made, modification, equivalent replacement, Improve etc., it should all be included in the protection scope of the present invention.