CN111138229A - Preparation and application of beneficial trichoderma and bacillus coated fertilizer - Google Patents
Preparation and application of beneficial trichoderma and bacillus coated fertilizer Download PDFInfo
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- CN111138229A CN111138229A CN202010014574.XA CN202010014574A CN111138229A CN 111138229 A CN111138229 A CN 111138229A CN 202010014574 A CN202010014574 A CN 202010014574A CN 111138229 A CN111138229 A CN 111138229A
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- 239000003337 fertilizer Substances 0.000 title claims abstract description 99
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 241000223259 Trichoderma Species 0.000 title abstract description 26
- 241000193830 Bacillus <bacterium> Species 0.000 title abstract description 15
- 230000009286 beneficial effect Effects 0.000 title abstract description 8
- 241000894006 Bacteria Species 0.000 claims abstract description 38
- 239000000843 powder Substances 0.000 claims abstract description 33
- 240000008042 Zea mays Species 0.000 claims abstract description 28
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 28
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 22
- 229910052939 potassium sulfate Inorganic materials 0.000 claims abstract description 18
- -1 potassium sulfate compound Chemical class 0.000 claims abstract description 18
- 235000011151 potassium sulphates Nutrition 0.000 claims abstract description 18
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 13
- 238000005303 weighing Methods 0.000 claims abstract description 5
- 239000000203 mixture Substances 0.000 claims abstract description 3
- 230000000443 biocontrol Effects 0.000 claims description 38
- 239000011248 coating agent Substances 0.000 claims description 30
- 241001460073 Trichoderma asperellum Species 0.000 claims description 26
- 238000000855 fermentation Methods 0.000 claims description 23
- 230000004151 fermentation Effects 0.000 claims description 23
- 238000000576 coating method Methods 0.000 claims description 18
- 150000001875 compounds Chemical class 0.000 claims description 17
- 239000002245 particle Substances 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 15
- 238000002156 mixing Methods 0.000 claims description 11
- 238000001694 spray drying Methods 0.000 claims description 11
- 230000001580 bacterial effect Effects 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- 239000008187 granular material Substances 0.000 claims description 6
- 239000007921 spray Substances 0.000 claims description 6
- 229920000858 Cyclodextrin Polymers 0.000 claims description 5
- 239000001116 FEMA 4028 Substances 0.000 claims description 5
- 235000010489 acacia gum Nutrition 0.000 claims description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 5
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims description 5
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims description 5
- 229960004853 betadex Drugs 0.000 claims description 5
- 239000007853 buffer solution Substances 0.000 claims description 5
- 229920001353 Dextrin Polymers 0.000 claims description 4
- 239000004375 Dextrin Substances 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 4
- 239000001785 acacia senegal l. willd gum Substances 0.000 claims description 4
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims description 4
- 239000001768 carboxy methyl cellulose Substances 0.000 claims description 4
- 235000019425 dextrin Nutrition 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 229920000609 methyl cellulose Polymers 0.000 claims description 4
- 239000001923 methylcellulose Substances 0.000 claims description 4
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims description 4
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 2
- 235000009973 maize Nutrition 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 229910052700 potassium Inorganic materials 0.000 claims description 2
- 238000005507 spraying Methods 0.000 claims 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 abstract description 26
- 235000005822 corn Nutrition 0.000 abstract description 26
- 230000000694 effects Effects 0.000 abstract description 10
- 239000000126 substance Substances 0.000 abstract description 9
- 201000010099 disease Diseases 0.000 abstract description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 8
- 241000207965 Acanthaceae Species 0.000 abstract 1
- 239000012467 final product Substances 0.000 abstract 1
- 238000011282 treatment Methods 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 238000009331 sowing Methods 0.000 description 7
- 241000223195 Fusarium graminearum Species 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 239000003124 biologic agent Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 3
- 241000223218 Fusarium Species 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 241001112741 Bacillaceae Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 241000223194 Fusarium culmorum Species 0.000 description 2
- 241000223221 Fusarium oxysporum Species 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 241000193833 Bacillales Species 0.000 description 1
- 208000003643 Callosities Diseases 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 239000005781 Fludioxonil Substances 0.000 description 1
- 241000233732 Fusarium verticillioides Species 0.000 description 1
- 241001274282 Fusarium zeae Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 239000005808 Metalaxyl-M Substances 0.000 description 1
- 241000219000 Populus Species 0.000 description 1
- 241000233639 Pythium Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 241000082085 Verticillium <Phyllachorales> Species 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000003625 amylolytic effect Effects 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000009274 differential gene expression Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- MUJOIMFVNIBMKC-UHFFFAOYSA-N fludioxonil Chemical compound C=12OC(F)(F)OC2=CC=CC=1C1=CNC=C1C#N MUJOIMFVNIBMKC-UHFFFAOYSA-N 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- ZQEIXNIJLIKNTD-GFCCVEGCSA-N metalaxyl-M Chemical compound COCC(=O)N([C@H](C)C(=O)OC)C1=C(C)C=CC=C1C ZQEIXNIJLIKNTD-GFCCVEGCSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 239000004563 wettable powder Substances 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Pest Control & Pesticides (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Fertilizers (AREA)
Abstract
本发明公开了一种益菌化的木霉菌和芽孢杆菌包衣化肥的制备及其应用；该方法包含以下操作步骤：(1)制备耐高温棘孢木霉菌粉剂；(2)制备解淀粉芽孢杆菌粉剂；(3)称取硫酸钾复合肥，N‑P2O5‑K2O 14‑16‑15；(4)步骤(3)中加入0.18％的肥料多功能包裹剂S1；(5)将硫酸钾复合肥和多功能包裹剂包裹均匀后，加入步骤(2)制得的解淀粉芽孢杆菌；(6)加入步骤(1)制得的棘孢木霉菌粉剂，混合均匀，即可。本发明制备的具生物防治功能的木霉菌和芽孢杆菌包衣化肥对玉米苗枯病、穗腐病和茎腐病和叶斑病等作物病害有较好的防治效果，同时提高化肥利用效率，促进玉米生长、提高玉米产量。The invention discloses a preparation and application of beneficial bacteria-enhanced Trichoderma and Bacillus-coated fertilizers; the method comprises the following operation steps: (1) preparing high-temperature resistant Trichoderma acanthus powder; (2) preparing amyloliquefaciens spores Bacillus powder; (3) take by weighing potassium sulfate compound fertilizer, N-P 2 O 5 -K 2 O 14-16-15; (4) in step (3), add 0.18% fertilizer multi-functional wrapping agent S1; (5) ) After the potassium sulfate compound fertilizer and the multifunctional wrapping agent are evenly wrapped, add the Bacillus amyloliquefaciens obtained in step (2); (6) add the Trichoderma aculeatus powder obtained in step (1), mix well, and get final product . The Trichoderma and Bacillus-coated chemical fertilizers with biological control functions prepared by the invention have better control effects on crop diseases such as corn seedling blight, ear rot, stem rot and leaf spot, and simultaneously improve the utilization efficiency of chemical fertilizers. Promote corn growth and increase corn yield.
Description
Technical Field
The invention relates to a bacterium benefiting technology of two biocontrol bacterium (trichoderma and bacillus) coating compound fertilizers and application thereof in corn disease prevention and yield increase; in particular to preparation and application of a beneficial trichoderma and bacillus coated fertilizer.
Background
In the production of corn, various soil-borne diseases are frequently encountered, such as corn seedling blight, stem rot, sheath blight, ear rot and the like. The main pathogenic bacteria of the maize seedling blight are Fusarium verticillium (Sacc.) Nirenberg, the morbidity of a common field is 10-50%, the seedlings of a heavily diseased field are often deficient by 3-5, and serious seedlings damage seeds and seriously affect the yield. The corn ear rot pathogenic bacteria mainly comprise Fusarium graminearum (compound species) and Fusarium verticillioides (Fusarium graminearum), and the like, and are mainly harmful to corn ear grains, so that direct yield reduction and toxin pollution are caused, the annual incidence rate of the re-transmitted corn ear rot is 30-40%, the incidence rate of susceptible varieties is more than 50%, and the yield loss is as high as 30-40%. The corn stalk rot is mainly caused by fusarium graminearum and Pythium spp, the morbidity of general fields is 10-20%, the serious field disease planting rate reaches 50-60%, the yield per mu is reduced by 10-30%, and the yield of serious fields is reduced by 50%, so that lodging and mechanical harvesting difficulty are caused. The soil-borne diseases do not have ideal fertilizer control technology, and biocontrol bacteria, such as trichoderma granules, can achieve control effects of over 60 percent when being mixed with chemical fertilizers before sowing, however, the mixed application technology of the trichoderma granules and the chemical fertilizers still has the defects of labor and time consumption, and the trichoderma is sensitive to stress-tolerant factors such as high temperature, so that how to coat the biocontrol bacteria on the surface of the fertilizer to form fertilizer particles beneficial to fertilization is realized, and the aims of efficiently controlling diseases by the synergism of the biocontrol microorganisms and the fertilizer, improving the utilization efficiency of the fertilizer and increasing the yield are fulfilled. The technology needs to overcome the following problems: (1) biocontrol bacteria need to tolerate the influence of high temperature in the coating drying process on the biocontrol bacteria activity; (2) the biocontrol bacteria need to endure the stress of the fertilizer per se and keep higher reproductive activity; (3) the difficult problem of easy caking in the processing engineering of the probiotic chemical fertilizer needs to be solved. Therefore, the invention selects and screens biocontrol trichoderma and bacillus amyloliquefaciens with high yield of stress-resistant spores, develops a novel microorganism-fertilizer composite preparation with a biocontrol function by a high-temperature drying resistant technology and a fertilizer stress resistant technology of a beneficial fertilizer, and realizes the upgrading and updating of traditional fertilizer varieties.
The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
Disclosure of Invention
The invention aims to provide preparation and application of a probiotic coated fertilizer containing trichoderma and bacillus, and the preparation method adopts a high-temperature resistant technology to prepare the biocontrol coated fertilizer by taking Arabic gum and β -cyclodextrin as a trichoderma high-temperature resistant protective agent in a ratio and combining a mode of low-temperature drying at the air inlet temperature of 115 ℃ and the air outlet temperature of 65 ℃.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
the invention relates to a preparation method of biocontrol bacteria coated fertilizer granules, which comprises the following steps:
s1, preparing high-temperature resistant trichoderma asperellum powder;
s2, preparing bacillus amyloliquefaciens powder;
s3, weighing the potassium sulfate compound fertilizer, adding the potassium sulfate compound fertilizer into a coating machine, and adjusting the rotating speed and the temperature of the coating machine;
s4, adding 0.18% (relative to the potassium sulfate compound fertilizer) of multifunctional fertilizer wrapping agent after the surface of the potassium sulfate compound fertilizer in the step S3 is raised to a preset temperature;
s5, when the surface of the potassium sulfate compound fertilizer is uniformly coated by the multifunctional fertilizer coating agent in the step S4, 4-5% (relative to the potassium sulfate compound fertilizer) of the bacillus amyloliquefaciens powder is added;
and S6, stopping heating after the particles are uniformly wrapped in the step S5, adding 4-5% of high-temperature resistant trichoderma asperellum powder when the surface temperature of the particles is reduced to be lower than the preset temperature in the step S5, and obtaining the biocontrol bacteria coated fertilizer particles after uniform wrapping.
Further, in step S1, the preparation of the high temperature resistant trichoderma asperellum powder comprises the following steps:
a1, inoculating the activated trichoderma asperellum strain into a PD culture medium, and culturing at 28 ℃ for 3-5 d to prepare a seed solution; adding the seed solution into a fermentation tank according to a proportion, and fermenting for 4-6 days at 25-28 ℃ to obtain fermentation liquor; centrifuging, removing supernatant and reserving bacterial pulp;
a2, uniformly mixing PBS buffer solution with the pH value of 5.5-6.0, 2-5% of the bacterial pulp, 5-10% of Arabic gum, 5-10% of β -cyclodextrin, 7-10% of NNO and 1-5% of ammonium sulfate, and then spray-drying to obtain the mixture with the concentration of 1-5 multiplied by 107CFU/g of Trichoderma asperellum powder.
Preferably, the activated trichoderma asperellum strain is inoculated into a PD culture medium and cultured for 3-5 d at 28 ℃ to prepare a seed solution; adding the seed solution into a fermentation tank in proportion, and fermenting for 4-6 days at 28 ℃ to obtain fermentation liquor; centrifuging, removing supernatant and reserving bacterial pulp;
mixing PBS buffer solution with pH of 5.8, 2% of the bacterial pulp, 5% of Arabic gum, 5% of β -cyclodextrin, 7% of NNO and 1% of ammonium sulfate uniformly, and spray drying to obtain the product with concentration of 5 × 107CFU/g of Trichoderma asperellum powder.
Further, in the step A2, the spray drying adopts a low-temperature drying mode that the inlet air temperature is 115-130 ℃ and the outlet air temperature is 65-80 ℃, and the spray rate of the atomizer is 300-350 Hz. Preferably, the spray drying adopts a low-temperature drying mode of air inlet temperature of 115 ℃ and air outlet temperature of 65 ℃, and the spray rate of the atomizer is 300 Hz.
Further, in step S2, the preparation of the bacillus amyloliquefaciens powder comprises the following steps:
b1, culturing the bacillus amyloliquefaciens strain at 37 ℃ for 1d to prepare a seed solution, adding the seed solution into a fermentation tank in proportion, and culturing at 30 ℃ for 48-60 h to obtain a fermentation liquid;
b2, adding 5-10% of diatomite, 4-10% of sodium carboxymethylcellulose, 5-6% of sodium dodecyl sulfate, 2-5% of methylcellulose and 0.1% of dextrin into the fermentation liquor, uniformly mixing, and spray-drying to obtain the product with the concentration of 2.3 x 1010CFU/g Bacillus amyloliquefaciens powder.
Preferably, the bacillus amyloliquefaciens strain is cultured for 1d at 37 ℃ to prepare a seed solution, the seed solution is added into a fermentation tank according to a proportion, and the fermentation liquid is obtained after the seed solution is cultured for 60 hours at 30 ℃; adding 10% of diatomite, 4% of sodium carboxymethylcellulose, 6% of sodium dodecyl sulfate, 2% of methylcellulose and 0.1% of dextrin into the fermentation liquor, uniformly mixing, and spray-drying to obtain the product with the concentration of 2.3 × 1010CFU/g Bacillus amyloliquefaciens powder.
Further, in the step B2, the air inlet temperature is 170-180 ℃, the air outlet temperature is 60-80 ℃, and the spray rate of the atomizer is 300-350 Hz. Preferably, the spray drying adopts the air inlet temperature of 170 ℃, the air outlet temperature of 60 ℃ and the spray rate of the atomizer of 300 Hz.
Further, in step S3, the rotation speed of the coating machine is 20-40 r/min, and the temperature is 45-60 ℃. Preferably, the rotating speed of the coating machine is 40r/min, and the temperature is 60 ℃.
Further, in step S3, the potassium sulfate compound fertilizer is a potassium sulfate compound fertilizer N-P2O5-K2The contents of O and N, P, K are respectively 14%, 16% and 15%, and the total nutrient of the compound fertilizer is more than or equal to 45%.
Further, in step S4, the preset temperature is 45 to 60 ℃.
Further, in step S6, the predetermined temperature is 45 to 50 ℃.
Further, in step S4, the multifunctional fertilizer coating agent is a multifunctional fertilizer coating agent that does not inhibit the growth of trichoderma asperellum.
The invention also relates to application of the biocontrol bacteria-coated fertilizer granules prepared by the preparation method in preventing and treating corn seedling blight, ear rot or stem rot.
The application is specifically implemented by furrow application or hole application of the biocontrol bacterium-coated fertilizer particles before sowing, wherein the amount of the biocontrol bacterium-coated fertilizer particles is 30 kg/mu.
Compared with the prior art, the invention has the following beneficial effects:
1) the biocontrol bacteria coated fertilizer prepared by the invention has better control effects on fusarium oxysporum f.sp.zeae, fusarium graminearum ear rot, fusarium culmorum rot and the like;
2) the biocontrol bacteria coated fertilizer prepared by the invention not only provides trace elements required by corn growth, but also provides metabolites which are generated by bacillus amyloliquefaciens and trichoderma asperellum and can promote corn growth and improve disease resistance;
3) the application of the biocontrol bacteria-coated fertilizer has a certain synergistic effect on improving the yield of the corn.
Detailed Description
The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that it would be apparent to those skilled in the art that several modifications and improvements can be made without departing from the inventive concept. All falling within the scope of the present invention.
Experimental materials: in the embodiment, the compound fertilizer is a potassium sulfate compound fertilizer purchased from the market; the trichoderma asperellum powder and the bacillus amyloliquefaciens powder are produced by the institute of microorganisms in the agricultural environment of Shanghai university of transportation; the multifunctional fertilizer wrapping agents S1 and S2 are purchased from HiCOL BRL101 and HICOL YL201, Hubei Fubang science and technology Co., Ltd; the multifunctional wrapping agent FB is purchased from Suzhou Fengbang biotechnology limited and has the model number of FB-W01.
Trichoderma asperellum (Trichoderma asperellum) is a species of Trichoderma (Trichoderma), and has antagonistic biocontrol bacteria against various pathogenic bacteria, and the distribution is very wide. They belong to the subdivision Deuteromycotina (Deuteromycina), class Hyphomycetes (Hyphomycetes), order Hyphomycetales (Hyphomycetales), family Hyphomycetaceae (Hyphomycetae) according to the classical classification system. Bacillus is a type of biocontrol bacterium that produces a range of metabolites that inhibit fungal and bacterial activity during growth. They belong to the kingdom of Bacteria (Bacteria), the phylum Firmicutes (Firmicutes), the class of Bacillaceae (Bacillus), the order of Bacillales (Bacillus), the family of Bacillaceae (Bacillus), the genus Bacillus (Bacillus). The following examples relate to T.asperellum GDFS1009 and B.amyloliquefaciens 1841 (these two strains are published in the International journal "Karuppiah V, Li T, Vallikkannu M, et al. Co-culture of Trichoderma asperellum GDFS1009 and Bacillus amyloliquefaciens 1841 uses differential gene expression and differentiation in the white growth and biocontrol activity [ J ]. Frondiers microbiology,2019,10: 1068.).
Example 1
The embodiment relates to a screening method of a coating material of a biocontrol bacteria coated fertilizer, which comprises the following operation steps:
(1) weighing the multifunctional wrapping agent, heating in a metal bath at 60 ℃ for 30min, wherein the raw materials are as follows: the multifunctional coating agent comprises S1 multifunctional coating agent, S2 multifunctional coating agent, FB multifunctional coating agent and FB + 2% CMC-Na solution.
(2) And (3) respectively taking 100 mu L of the raw materials in the step (1), uniformly mixing with 10mL of PDA culture medium (the PDA culture medium comprises 200g of potatoes, 20g of glucose, 15g of agar and 1L of pure water, and the pH value is natural), and respectively preparing the plates with the wrapping materials.
(3) And (2) respectively taking 100 mu L of the raw materials in the step (1), uniformly mixing with 10mL of LB solid medium (10 g of tryptone, 5g of yeast extract powder and 10g of NaCl, pH7.0, fixing the volume to 1L, and adding 15g of agar), and respectively preparing the plates with the wrapping materials.
(4) Edges of the trichoderma colonies that had been activated on the PDA medium were punched out with a sterilization punch (phi. ═ 5mm), and inoculated on the plates in step (2), respectively, and observed day by day.
(5) Single colonies of spores that had been activated on LB solid medium were picked, streaked on the plates in step (3), respectively, and observed day by day.
The raw material which can not inhibit the growth of trichoderma asperellum obtained in the embodiment can be used for a wrapping material of a compound fertilizer, in the embodiment, the bacteriostasis rate of the multifunctional wrapping agent S1 on trichoderma is 0%, the bacteriostasis rate of the multifunctional wrapping agent S2 on trichoderma is 43.10%, the bacteriostasis rate of the multifunctional wrapping agent FB on trichoderma is 85.96%, and the bacteriostasis rate of the FB + 2% CMC-NA solution on trichoderma is 88.50%. Spores grew normally on the plates containing the multifunctional wrapping agent S1, and none of the other treatments could grow. Therefore, the S1 multifunctional coating agent can be used for coating compound fertilizers.
Example 2
The embodiment relates to a preparation method of trichoderma asperellum powder
(1) Activating trichoderma asperellum on a PDA culture medium for about 5 days;
(2) and inoculating the activated strain into 300mL/500mL PD culture medium, and culturing at 28 ℃ at 200r/min for 3-5 d to finish the preparation of seed liquid.
(3) Adding the seed liquid into a fermentation tank according to the proportion of 0.3 percent, and fermenting according to the following formula: the formula of the fermentation medium is 50g/L corn flour, 3.82g/L potassium dihydrogen phosphate, 1.42g/L sodium nitrate, 1.1g/L ammonium sulfate, 1g/L sodium chloride, 0.5g/L magnesium sulfate heptahydrate, 0.0075g/L ferrous sulfate heptahydrate, 0.0025g/L manganese sulfate and 0.002g/L zinc sulfate, water is added, 200L water is added, and fermentation is carried out for 4-6 d to obtain fermentation liquor.
(4) Centrifuging the fermentation liquid at 12000rpm for 30min, removing supernatant to obtain bacterial pulp, mixing PBS buffer solution with pH of 5.8, 2% bacterial pulp, 5% acacia gum, 5% β -cyclodextrin, 7% NNO and 1% ammonium sulfate (the percentage content of each substance is relative to the volume of PBS buffer solution), drying with spray drier at inlet air temperature of 115 deg.C and outlet air temperature of 65 deg.C to obtain concentration of 5 × 107CFU/mL of Trichoderma asperellum powder.
Example 3
The embodiment relates to a preparation method of a biocontrol bacteria coated fertilizer, which comprises the following operations:
(1) preparing high-temperature resistant trichoderma asperellum powder; (same as example 2)
(2) Weighing potassium sulfate compound fertilizer, adding into a coating machine, and adjusting the rotating speed of the coating machine to 40r/min and the temperature to 60 ℃;
(3) after the compound fertilizer in the step (2) is raised to a preset temperature, adding 0.18 percent (relative to the weight of the compound fertilizer) of multifunctional fertilizer wrapping agent S1;
(4) when the multifunctional fertilizer coating agent in the step (3) uniformly coats the surface of the compound fertilizer, adding 4% (relative to the weight of the compound fertilizer) of bacillus amyloliquefaciens powder;
(5) and (3) stopping heating after uniform coating, adding 4 percent (relative to the weight of the compound fertilizer) of trichoderma asperellum powder when the surface temperature of the particles in the step (4) is reduced to be below 45 ℃ (the temperature range clearly indicated by the worship), and obtaining the disease-resistant and beneficial bacterial fertilizer after uniform coating. The fertilizer obtained in this embodiment is the biocontrol bacteria coated fertilizer.
The preparation method of the bacillus amyloliquefaciens powder refers to the development of bacillus amyloliquefaciens X17 wettable powder which is published in China biological control newspaper of Qian Yixin, congyuan, Roth Kjen, Gunn Kun, Lebangxing, Zhang.
The amylolytic spores are cultured at 37 ℃ for 1d to prepare seed liquid, the seed liquid is added into a fermentation tank according to the proportion of 0.3 percent, and the seed liquid is fermented according to the following formula: 20g of yeast extract powder, 20g of molasses, 10g of corn protein powder, 0.5g of monopotassium phosphate (KH2PO4), 0.5g of magnesium sulfate (MgSO4 & 7H2O), 0.3g of sodium chloride (NaCl), 3g of calcium carbonate (CaCO3) and 0.5g of defoaming agent, adding 100L of water, and culturing at 30 ℃ for 60 hours to obtain a fermentation liquid; adding 10% of diatomite, 4% of sodium carboxymethylcellulose, 6% of sodium dodecyl sulfate, 2% of methylcellulose and 0.1% of dextrin (the percentage content of each substance is relative to the volume of the fermentation liquor), uniformly mixing, and spray-drying to obtain the product with the concentration of 2.3 × 1010CFU/g Bacillus amyloliquefaciens powder.
Example 4
The embodiment relates to the application of the biocontrol bacteria coated fertilizer prepared in the embodiment 3 in preventing and treating fusarium corn diseases:
set 3 treatments:
(1) biocontrol bacteria coating fertilizer, 30 kg/mu;
(2) 30 kg/mu of uncoated compound fertilizer;
(3) in contrast to CK, the seed treated with metalaxyl-M fludioxonil corn grain seed coating agent was first reached;
the method is applied to the seriously ill corn field of the tile house in 2017 and 2018 respectively.
The test sites in 2017 are arranged in the Yangjia village in the Yangjia, the Sun-countryside village in the Wakjang city and the three villages in the Wakjang city, and the areas of the three villages are 10 mu each. Performing hole application on the treatments (1) and (2) before sowing the corns, and performing conventional treatment at the later stage. As can be seen from the results (tables 1 and 2), the biocontrol bacteria-coated fertilizer has good control effects on fusarium oxysporum f.zeae, fusarium graminearum ear rot and fusarium culmorum stem rot of corn, and the control effects in 3 test fields are all more than or equal to 40.00%. The biocontrol bacterium-coated fertilizer has the effect of increasing the yield of the corn, and the yield is increased by more than 11.00 percent. (the yield increase and control effect data in tables 1 and 2 were calculated using the treatment (3) as a control group)
TABLE 1
TABLE 2
Example 5
The embodiment relates to the application of the biocontrol bacteria coated fertilizer prepared in the embodiment 3 in preventing and treating fusarium corn diseases:
set 3 treatments:
(1) biocontrol bacteria coating fertilizer, 30 kg/mu;
(2) 30 kg/mu of uncoated compound fertilizer;
(3) in contrast to CK, seeds were not coated, fertilizer was not applied;
the test site in 2018 is set in the Malay Chenghui of Yanjiacun of Yang Home of the Populus of Wakuang shop, and the area is 10 mu each. Performing hole sowing before sowing the corn, performing conventional post-treatment, arranging the treated and contrasted plants in the same field with 10 mu each, setting two fields, and performing hole sowing of the treated plants (1) and (2) respectively before sowing the corn. From the results (table 3), it can be seen that treatment (1) significantly improved the control of corn head rot, and both the control in the mary home and maryan home were superior to the uncoated compound fertilizer treatment. The yield increase of the treatment (1) is more than 20 percent, and is obviously better than that of the treatment (2). (the data on the yield increase and control effect in Table 3 were obtained by calculation using the treatment (3) as a control group)
TABLE 3
The high-temperature protective agent added in the preparation method of the trichoderma can quickly dry the microbial inoculum, protect the trichoderma from high-temperature loss of a coating machine, greatly shorten the preparation time limit of the trichoderma biofertilizer and improve the productivity; compared with the traditional method that the biological agent is mixed with the fertilizer for application after being prepared, the trichoderma and the bacillus are uniformly attached to the surface of the fertilizer, so that the using amount of the biological agent can be reduced, and the using efficiency is obviously improved; in addition, the traditional process of mixing and applying the biological agent after preparation with the chemical fertilizer can not eliminate the influence of the compound fertilizer on the growth of microorganisms, the multifunctional wrapping agent used by the invention can prevent the biological agent from directly contacting with the chemical fertilizer, reduce the lethal effect of urea and the like contained in the compound fertilizer on trichoderma, and obviously improve the survival rate of viable bacteria of the microbial agent.
The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.
Claims (10)
1. A preparation method of biocontrol bacteria coated fertilizer particles is characterized by comprising the following steps:
s1, preparing high-temperature resistant trichoderma asperellum powder;
s2, preparing bacillus amyloliquefaciens powder;
s3, weighing the potassium sulfate compound fertilizer, adding the potassium sulfate compound fertilizer into a coating machine, and adjusting the rotating speed and the temperature of the coating machine;
s4, adding 0.18% of multifunctional fertilizer wrapping agent after the surface of the potassium sulfate compound fertilizer in the step S3 is raised to a preset temperature;
s5, when the multifunctional fertilizer wrapping agent uniformly wraps the surface of the potassium sulfate compound fertilizer in the step S4, adding 4-5% of bacillus amyloliquefaciens powder;
and S6, stopping heating after the particles are uniformly wrapped in the step S5, adding 4-5% of high-temperature resistant trichoderma asperellum powder when the surface temperature of the particles is reduced to be lower than the preset temperature in the step S5, and obtaining the biocontrol bacteria coated fertilizer particles after uniform wrapping.
2. The method of claim 1, wherein in step S1, the preparation of the high temperature resistant trichoderma asperellum powder comprises the steps of:
a1, inoculating the activated trichoderma asperellum strain into a PD culture medium, and culturing at 28 ℃ for 3-5 d to prepare a seed solution; adding the seed solution into a fermentation tank according to a proportion, and fermenting for 4-6 days at 25-28 ℃ to obtain fermentation liquor; centrifuging, removing supernatant and reserving bacterial pulp;
a2, uniformly mixing PBS buffer solution with the pH value of 5.5-6.0, 2-5% of the bacterial pulp, 5-10% of Arabic gum, 5-10% of β -cyclodextrin, 7-10% of NNO and 1-5% of ammonium sulfate, and then spray-drying to obtain the mixture with the concentration of 1-5 multiplied by 107CFU/g of Trichoderma asperellum powder.
3. The method for preparing biocontrol bacteria-coated fertilizer particles as claimed in claim 2, wherein in step A2, the spray drying is performed by using a low-temperature drying mode with the inlet air temperature of 115-130 ℃ and the outlet air temperature of 65-80 ℃, and the spray rate of the atomizer is 300-350 Hz.
4. The method of claim 1, wherein the step of preparing the powder of Bacillus amyloliquefaciens at step S2 comprises the steps of:
b1, culturing the bacillus amyloliquefaciens strain at 37 ℃ for 1d to prepare a seed solution, adding the seed solution into a fermentation tank in proportion, and culturing at 30 ℃ for 48-60 h to obtain a fermentation liquid;
b2, adding 5-10% of diatomite, 4-10% of sodium carboxymethylcellulose, 5-6% of sodium dodecyl sulfate, 2-5% of methylcellulose and 0.1% of dextrin into the fermentation liquor, uniformly mixing, and then spray-dryingTo obtain a concentration of 2.3X 1010CFU/g Bacillus amyloliquefaciens powder.
5. The method for preparing biocontrol bacteria-coated fertilizer particles as claimed in claim 4, wherein in step B2, the spray drying adopts the air inlet temperature of 170-180 ℃, the air outlet temperature of 60-80 ℃ and the spraying rate of the atomizer of 300-350 Hz.
6. The method for preparing biocontrol bacteria-coated fertilizer particles as claimed in claim 1, wherein in step S3, the coating machine is rotated at a speed of 20-40 r/min and the temperature is 45-60 ℃.
7. The method for preparing biocontrol bacteria-coated fertilizer granules of claim 1, wherein in step S3, the potassium sulfate compound fertilizer is potassium sulfate compound fertilizer N-P2O5-K2The contents of O and N, P, K are respectively 14%, 16% and 15%, and the total nutrient of the compound fertilizer is more than or equal to 45%.
8. The method for preparing biocontrol bacteria-coated fertilizer particles as claimed in claim 1, wherein in step S4, the preset temperature is 45-60 ℃; in step S6, the predetermined temperature is 45-50 ℃.
9. The method of claim 1, wherein in step S4, the multifunctional fertilizer coating agent is a multifunctional fertilizer coating agent that does not inhibit the growth of Trichoderma asperellum.
10. The application of the biocontrol bacteria-coated fertilizer granules prepared by the preparation method of any one of claims 1-9 in preventing and treating maize seedling blight, ear rot or stem rot.
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