CN103039439B - Seed coating agent of Trichoderma fungi and preparation method thereof - Google Patents
Seed coating agent of Trichoderma fungi and preparation method thereof Download PDFInfo
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- CN103039439B CN103039439B CN201110306008.7A CN201110306008A CN103039439B CN 103039439 B CN103039439 B CN 103039439B CN 201110306008 A CN201110306008 A CN 201110306008A CN 103039439 B CN103039439 B CN 103039439B
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Abstract
The invention discloses a pesticide seed coating agent, a seed coating agent film forming material and a preparation method of the seed coating agent. According to the invention, active chlamydospore of bacteria fermentation broth is added into a film forming material prepared from polyethylene glycol, xanthan gum and methyl cellulose, and then the seeds are wrapped. The seed coating agent is uniform in appearance and not easy to drop, and does not influence budding rate, wherein number of the Trichoderma chlamydospore on each seed is not less than 10<4> CFU; and the seed coating agent can effectively protect seeds in a germination stage.
Description
Technical field
The present invention relates to a kind of pesticide seed coating and preparation method thereof, particularly relate to a kind of wooden mould fungi seed coat agent and preparation method thereof.
Background technology
Seed is the important guarantee of crops high yield high-quality, but potential pathogen usually affects seed germination in the pathogen that seed itself carries or soil, current seed treatment is extremely important, effectively and one of plant protection means easily, wherein seed coat agent is owing to having the advantages such as easy to use, cost accounting is low, lasting medicine, is very applicable to extensive pesticide producing.Seed coat agent is coated on the surface of the seed, and formed and have the protective layer of some strength and permeability, its preferred component comprises insecticide, bactericide, micro-fertilizer, auxin etc., can effectively preventing soil insect, plant pass, soil-borne disease.Current seed coat agent is to be widely used in the plant protection process of many crops such as soybean, corn, wheat, paddy rice, potato, cotton, capsicum.
Tradition seed coat agent preferred component is chemical pesticide, and as carbendazim, Baijunling, tmtd, Imidacloprid etc., suspension seed-coating agent disclosed in patent CN1444853A, adopts tmtd and olefin conversion 13:2 proportioning to prepare seed coat agent; Suspension seed-coating agent disclosed in patent CN101147489A, employing be the composite mode of prochloraz and Imidacloprid; Corn suspension seed coating disclosed in patent CN101926346A, then what adopt is that carbofuran and thiophanate-methyl are composite; Dressing agent for rice disclosed in patent CN1429485A, adopts ethiprole and Imidacloprid to carry out composite; Potato seed-coating agent disclosed in patent CN101238814A, adopts Imidacloprid and metalaxyl to carry out composite; Cucumber seed coat agent disclosed in patent CN100563442C, adopts KH
2pO
4, CuSO
4composite with paclobutrazol.But many degree of chemical pesticide are produced and are used, ecotope is damaged, and residue of pesticide are detrimental to health especially, simultaneously beneficial bacterium is killed in desinsection process in sterilization by chemical pesticide together with beneficial insect, and Long-Time Service also can formula insect and pathogen develop immunity to drugs.The negative effect that chemical pesticide brings has caused global extensive concern, and various countries have in succession put into effect the forbidding of chemical pesticide, first used regulation.
Biopesticide because of its in degraded, pollute little, environment compatibility good, not easily produce the advantages such as resistance to person poultry safety and disease and insect, started progressively substituting chemical pesticide.Seed coat agent prepared by current biopesticide also has some patent reports, and patent CN100532538C discloses a kind of bacillus licheniformis seed coat agent, for preventing and treating soybean packing nematodosis and root rot; Patent CN100391345C discloses a kind of bacillus thuringiensis,Bt seed coat agent, has control efficiency to soybean packing nematodosis; The open coating agent for seed of patent CN1843118A for active ingredient, has the effect well walking quickly and keeping away muroid with the cycloheximide be separated in streptomyces griseus SPRI-201 culture; Seed coat agent disclosed in patent WO2009/126473A1, with multiple bacterium, fungies such as bacillus, basidiomycetes, chytrids for active component.
Filmogen is seed coat agent important component part, conventional synthesis macromolecule filming material has polyvinyl acetate, polyvinyl alcohol, polyacrylic etc., conventional natural polymer filmogen has cellulose and polysaccharide, as gelatin, sodium carboxymethylcellulose, gum Arabic, starch etc. have the material of cohesiveness and moulding property, disclosed in patent US20100285962A1, filmogen includes plant based fiber element and albumen; Seed coat agent disclosed in patent WO00/35277A1, component comprises polyvinyl acetate, methylcellulose, polyvinyl alcohol, viscose fiber and PVP(polyvinyl pyrrolidone) and glycan; Seed coat agent disclosed in patent CN100425632C adopts acrylic acid-acrylamide copolymer as filmogen; Film forming agent disclosed in patent CN1303864C adopts ethylene-vinyl acetate copolymer and polyvinyl alcohol to be filmogen; Film forming agent disclosed in patent CN101438701A adopts polyvinyl alcohol, oxidized starch-acrylamide copolymer is film forming agent.What other filmogen seed coat agent disclosed in outer patent CN1974629A adopted is that citric acid-ethylene glycol polyester is as filmogen; Seed coat agent disclosed in patent CN1265744C adopts animal blood dry powder as filmogen; Also inorganic carbonate is added in filmogen disclosed in patent EP2229808A1.But, still there is many defects in current many filmogens or filming performance is poor or environmental suitability is poor or cost is high.
Summary of the invention
The present invention's first object is to provide a kind of seed coat agent filmogen, filmogen good film-forming property of the present invention, difficult drop-off, shows evenly, can state and be widely used in various ecotope.
The present invention's second object is to provide a kind of seed coat agent comprising institute's filmogen, by spore dressing on crop seed, do not affect the vigor of seed, and plant growth and growth can be promoted, " microcosmic is ecological " in crop growth process is changed simultaneously, make around the rhizosphere of crop, have a large amount of Antagonistic Fungis to exist, to reach good disease resisting effect.
The present invention's the 3rd object is to provide a kind of method preparing described seed coat agent.
A kind of seed coat agent filmogen of the present invention, component comprises:
Glycerine 2 ~ 4 weight portion;
Polyethylene glycol 2 ~ 5 weight portion;
Xanthans 0.5 ~ 1 weight portion;
Methylcellulose 0.5 ~ 1 weight portion;
Tween 80 0.5 ~ 1 weight portion.
A kind of seed coat agent comprising described filmogen of the present invention, comprises bioactive ingredients and filmogen, and wherein, described filmogen component comprises:
Glycerine 2 ~ 4 weight portion;
Polyethylene glycol 2 ~ 5 weight portion;
Xanthans 0.5 ~ 1 weight portion;
Methylcellulose 0.5 ~ 1 weight portion;
Tween 80 0.5 ~ 1 weight portion.
Wherein, described bioactive ingredients comprises the chlamydospore of one or more bacterial classification, conidium or mycelium, and is preferably chlamydospore.
Wherein, described bacterial classification is one or more the mixture in wooden enzyme fungi, be preferably the center preservation of Trichoderma harzianum CGMCC No.5203(China Committee for Culture Collection of Microorganisms's common micro-organisms), Trichoderma harzianum CGMCC No.5202(China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation), the center preservation of trichoderma asperellum CGMCC No.5201(China Committee for Culture Collection of Microorganisms's common micro-organisms) in a kind of or several arbitrarily mixtures, above-mentioned three kinds of Trichodermas are separated by Wan Lihua bio tech ltd, Shanghai and obtain, and be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on August 31st, 2011, China Committee for Culture Collection of Microorganisms's common micro-organisms centre address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
According to a kind of preferred embodiment of seed coat agent of the present invention, warning colouration can also be added, and preferred water dissolubility Yihong uses as warning colouration; Its consumption is preferably 2 weight portions.
A preparation method for described seed coat agent, step comprises:
Step 1, strain fermentation, filters mycelia and obtains bioactive ingredients;
Step 2, the water-soluble solution of glycerine, polyethylene glycol, xanthans, methylcellulose and Tween 80, stirs and obtains filmogen;
Step 3, mixes described bioactive ingredients and filmogen;
Step 4, mixture step 3 obtained carries out wrapping up in kind of a process, dry.
Wherein, in step 1, described bioactive ingredients is diluted to 1.0 × 10
6cFU/ml.
Wherein, dry preferably dry 0.5 ~ 2h under 35 DEG C of conditions described in step 4.
Wherein, mixture described in step 4 and seed are preferably to carry out wrapping up according to volume ratio 1:25 and kind process.
By above-mentioned design, the present invention has following beneficial effect:
1. filmogen good film-forming property of the present invention, difficult drop-off, surface uniform, non-corrosiveness, adopt natural material to be main, good environmental adaptability.
2. seed coat agent of the present invention can with multiple bacterial classification or bacterial strain for bioactive ingredients, applied widely.Be convenient to store, transport and packaging simultaneously, and good stability, the storage life is long.
3. seed coat agent of the present invention is by reesei spores dressing on seed, does not affect seed vitality, and can change " microcosmic is ecological " in crop growth process, makes have a large amount of Antagonistic Fungi to exist around crop rhizosphere, can reach effective disease resisting effect.
4. in seed coat agent of the present invention, trichoderma strain content is high, therefore, has better prophylactic effect.
Embodiment
Seed coat agent filmogen component of the present invention comprises:
Glycerine 2 ~ 4 weight portion;
Polyethylene glycol 2 ~ 5 weight portion;
Xanthans 0.5 ~ 1 weight portion;
Methylcellulose 0.5 ~ 1 weight portion;
Tween 80 0.5 ~ 1 weight portion.
Can also comprise:
Warning colouration (preferred water dissolubility Yihong) 2 weight portions.
The present invention prepares the method for described seed coat agent, and step comprises:
Step 1, biological active component raw material, obtains spore zymotic fluid by fermentation, filters out mycelia, obtain bioactive ingredients;
Step 2, form filmogen by methylcellulose, polyethylene glycol, xanthans, glycerine and Tween 80, the water taken a morsel adds Tween 80 and polyethylene glycol, is placed on magnetic stirring apparatus and stirs, get glycerine, xanthans and methylcellulose again to add respectively, stir;
Step 3, mixes filmogen prepared by bioactive ingredients and step 2, now can also add warning colouration;
Step 4, preferably wraps up in kind of a process with volume ratio 1:25 ratio to seed, and after film forming agent is coated with seed, 35 DEG C are dried 0.5 ~ 2h.
Described bioactive ingredients is preferably the mixing of any one or a few bacterial strain in trichoderma harzianum CGMCC No.5202, trichoderma harzianum CGMCC No.5203, Trichoderma asperellum CGMCC No.5201 or bacterial strain fermentation liquor.
Below by specific embodiment the present invention described in detail and describe, to make better to understand the present invention, but the following embodiment remarkable scope of the invention.
embodiment 1
Strain fermentation prepares bioactive ingredients, and conventional solid can be adopted to ferment for fermentation process or conidium, chlamydospore or mycelium are prepared in liquid state fermentation, and being diluted to spore count is 1.0 × 10
6cFU/ml.
The water taken a morsel, adds the Tween 80 of 0.5g and 2g polyethylene glycol and is placed on magnetic stirring apparatus and stirs, then gets 4g glycerine, 1g xanthans and 0.5g methylcellulose and add respectively, and stir, 2g continues to stir in water-soluble Yihong.
Carry out wrapping up in kind of a process with volume ratio 1:20 ratio, after film forming agent is coated with seed, dry 1 ~ 2h for 35 DEG C.
embodiment 2
Strain liquid fermentation is for chlamydospore zymotic fluid, and 3 layers of filtered through gauze fall mycelia, are diluted to trichoderma chlamydospore numerical digit 1.0 × 10
6cFU/ml.Fermentation process is as follows:
Step 1, strain inoculation in medium, constant temperature culture;
Step 2, is transferred to culture fluid by the bacterial classification after cultivation and carries out Shaking culture, obtain conidium liquid;
Step 3, drops into culture fluid in fermentation tank, autoclave sterilization, then pours in fermentation tank by the conidium liquid obtained in step 2, and pressurization passes into air, ferments, obtains chlamydospore;
Described culture fluid component comprises:
| Broad bean powder: | 2wt% |
| Glucose: | 0.2wt% |
| Yeast extract: | 0.4wt% |
| Mixing Juice: | 3wt% |
| PH value: | 7.5 |
The water taken a morsel, adds the Tween 80 of 1g and 5g polyethylene glycol and is placed on magnetic stirring apparatus and stirs, then gets 3g glycerine, 0.5g xanthans and 1g methylcellulose and add respectively, and stir, 2g continues to stir in water-soluble Yihong.Seed coat agent is prepared with reference to method described in embodiment 1.
Detect through microscope 400 times of amplifying observations, seed coat agent prepared by said method of the present invention at normal temperatures (18 DEG C ~ 28 DEG C) shelf time (i.e. spore survival period) can reach more than 6 months, substantially prolongs the product shelf phase.
embodiment 3
Trichoderma CGMCC No.5203 fermentation is for chlamydospore zymotic fluid, and 3 layers of filtered through gauze fall mycelia, are diluted to trichoderma chlamydospore numerical digit 1.0 × 10
6cFU/ml.
The water taken a morsel, adds the Tween 80 of 0.5g and 2g polyethylene glycol and is placed on magnetic stirring apparatus and stirs, then gets 4g glycerine, 1g xanthans and 0.5g methylcellulose and add respectively, stirs, then adds 2g warning colouration and continue to stir.
With volume ratio 1:20 ratio, kind of a process is wrapped up in cucumber seeds, after film forming agent is coated with seed, dry 1 ~ 2h for 35 DEG C.
Pythium spp (
pythium spp), sickle-like bacteria (
fusurium spp), rhizoctonia (
rhizactonia spp) seed in sprouting can be infected, cause rotten kind, affect seed germination rate.Experimental result shows, and uses the root rot control efficiency in seedling stage of seed coat agent of the present invention to be measure seed coat agent after 91%, 6 each moons, every mould number>=1.7 × 10 of seed wood
4.
And Trichoderma known at present there is no method prepares wooden mould number and can reach 10
4the seed coat agent of content.
embodiment 4
Trichoderma CGMCC No.5201 and CGMCC No.5202 fermentation are for chlamydospore zymotic fluid, and 3 layers of filtered through gauze fall mycelia, are diluted to trichoderma chlamydospore numerical digit 1.0 × 10
6cFU/ml.
The water taken a morsel, adds the Tween 80 of 0.5g and 2g polyethylene glycol and is placed on magnetic stirring apparatus and stirs, then gets 4g glycerine, 0.5g xanthans and 0.5g methylcellulose and add respectively, stirs, then adds the water-soluble Yihong of 2g and continue to stir.
With volume ratio 1:20 ratio, kind of a process is wrapped up in cucumber seeds, after film forming agent is coated with seed, dry 1 ~ 2h for 35 DEG C.
Experimental result shows, and uses the root rot control efficiency in seedling stage of seed coat agent of the present invention to be measure seed coat agent after 93%, 6 each moons, every mould number>=1.1 × 10 of seed wood
4.
embodiment 5
Trichoderma CGMCC No.5201 fermentation is for chlamydospore zymotic fluid, and 3 layers of filtered through gauze fall mycelia, are diluted to trichoderma chlamydospore numerical digit 1.0 × 10
6cFU/ml.
The water taken a morsel, adds the Tween 80 of 0.5g and 2g polyethylene glycol and is placed on magnetic stirring apparatus and stirs, then gets 4g glycerine, 0.5g xanthans and 0.5g methylcellulose and add respectively, stir, then continues to stir in the water-soluble Yihong of 2g.
With volume ratio 1:20 ratio, kind of a process is wrapped up in cucumber seeds, after film forming agent is coated with seed, dry 1 ~ 2h for 35 DEG C.
Experimental result shows, and uses the root rot control efficiency in seedling stage of seed coat agent of the present invention to be measure seed coat agent after 93%, 6 each moons, every mould number>=1.3 × 10 of seed wood
4.
embodiment 6
Trichoderma CGMCC No.5201, CGMCC No.5202, CGMCC No.5203 fermentation are for chlamydospore zymotic fluid, and 3 layers of filtered through gauze fall mycelia, are diluted to trichoderma chlamydospore numerical digit 1.0 × 10
6cFU/ml.
The water taken a morsel, adds the Tween 80 of 0.5g and 2g polyethylene glycol and is placed on magnetic stirring apparatus and stirs, then gets 4g glycerine, 0.5g xanthans and 0.5g methylcellulose and add respectively, stirs, then adds the water-soluble Yihong of 2g and continue to stir.
With volume ratio 1:20 ratio, kind of a process is wrapped up in cucumber seeds, after film forming agent is coated with seed, dry 1 ~ 2h for 35 DEG C.
Experimental result shows, and uses the root rot control efficiency in seedling stage of seed coat agent of the present invention to be measure seed coat agent after 95%, 6 each moons, every mould number>=1.5 × 10 of seed wood
4.
When above-described embodiment illustrates that seed coat agent of the present invention take trichoderma chlamydospore as seed coat agent active component, effectively can suppress crop pest, therefore obvious effect of increasing production.
Also find by experiment, seed coat agent of the present invention is convenient to storage, transport and packaging, and by Trichoderma spore dressing on crop seed, do not affect the vigor of seed, and can promote that plant growth can also be grown, " microcosmic is ecological " in crop growth process is changed simultaneously, make around the rhizosphere of crop, have the mould Antagonistic Fungi that waits of a large amount of wood to exist, to reach good disease resisting effect.
According to the method for above-described embodiment, adopt trichoderma strain prevention root rot known at present, compared with most of known trichoderma strain, seed coat agent prepared by the above embodiment of the present invention, significantly can improve wooden mould content, thus prevention root rot effect in seedling stage is all greatly improved.
Detect through experiment, seed coat agent appearance uniform, the difficult drop-off prepared, do not affect bud ratio with filmogen of the present invention, every seed chlamydospore number can reach 10
4more than CFU, can carry out available protecting to germination period seed.
Be described in detail specific embodiments of the invention above, but it is just as example, the present invention is not restricted to specific embodiment described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and substituting also all among category of the present invention.Therefore, equalization conversion done without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.
Claims (7)
1. the mould fungi seed coat agent of wood, is characterized in that, comprise bioactive ingredients and filmogen, and wherein, described filmogen component comprises:
Described bioactive ingredients comprises chlamydospore or the conidium of one or more bacterial classification, and described bacterial classification is one or more the Mixed Microbes in trichoderma harzianum CGMCC No.5202, trichoderma harzianum CGMCC No.5203, Trichoderma asperellum CGMCCNo.5201.
2. seed coat agent according to claim 1, is characterized in that, component also comprises:
Warning colouration 2 weight portion.
3. a seed coat agent filmogen as claimed in claim 1, it is characterized in that, component comprises:
4. prepare a method for seed coat agent as claimed in claim 1, it is characterized in that, step comprises:
Step 1, strain fermentation, filter mycelia and obtain bioactive ingredients, described bioactive ingredients comprises chlamydospore or the conidium of one or more bacterial classification, and described bacterial classification is one or more the Mixed Microbes in trichoderma harzianum CGMCC No.5202, trichoderma harzianum CGMCC No.5203, Trichoderma asperellum CGMCC No.5201;
Step 2, the water-soluble solution of glycerine, polyethylene glycol, xanthans, methylcellulose and Tween 80, stirs and obtains filmogen;
Step 3, mixes described bioactive ingredients and filmogen;
Step 4, mixture step 3 obtained carries out wrapping up in kind of a process, dry.
5. method according to claim 4, is characterized in that, in step 1, described bioactive ingredients is diluted to 1.0 × 10
6cFU/ml.
6. method according to claim 4, is characterized in that, mixture described in step 4 and seed carry out wrapping up according to volume ratio 1:25 and kind to process.
7. method according to claim 4, is characterized in that, drying described in step 4 is dry 0.5 ~ 2h under 35 DEG C of conditions.
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| CN201110306008.7A CN103039439B (en) | 2011-10-11 | 2011-10-11 | Seed coating agent of Trichoderma fungi and preparation method thereof |
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| MX2018007932A (en) * | 2015-12-28 | 2019-02-26 | Novozymes Bioag As | Stable inoculant compositions and methods for producing same. |
| CN106172508B (en) * | 2016-07-11 | 2018-08-21 | 中国科学院昆明植物研究所 | A kind of Radix Notoginseng seedling root system biology coating agent and the preparation method and application thereof |
| CN106818887A (en) * | 2016-12-30 | 2017-06-13 | 新昌县派特普科技有限公司 | A kind of trichoderma fungi seed coat agent and preparation method thereof |
| CN107258824A (en) * | 2017-07-07 | 2017-10-20 | 云南农业大学 | A kind of biological coating agent for preventing and treating coix smut |
| CN109207381A (en) * | 2018-11-02 | 2019-01-15 | 河南省科学院生物研究所有限责任公司 | It is a kind of prevent and treat nematodiasis biocontrol bacterial strain SFC-3 and its application |
| CN109892327A (en) * | 2019-04-15 | 2019-06-18 | 微生物肥料技术研究推广中心 | A kind of preparation method and applications of microorganism seed coating |
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