JPH04353593A - Material for inoculation of microorganism - Google Patents
Material for inoculation of microorganismInfo
- Publication number
- JPH04353593A JPH04353593A JP3156006A JP15600691A JPH04353593A JP H04353593 A JPH04353593 A JP H04353593A JP 3156006 A JP3156006 A JP 3156006A JP 15600691 A JP15600691 A JP 15600691A JP H04353593 A JPH04353593 A JP H04353593A
- Authority
- JP
- Japan
- Prior art keywords
- soil
- microorganisms
- microorganism
- inoculation
- pva
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 244000005700 microbiome Species 0.000 title claims abstract description 69
- 239000000463 material Substances 0.000 title claims abstract description 24
- 238000011081 inoculation Methods 0.000 title abstract description 10
- 239000002689 soil Substances 0.000 claims abstract description 64
- 239000000126 substance Substances 0.000 claims abstract description 10
- 229920003169 water-soluble polymer Polymers 0.000 claims 2
- 239000004372 Polyvinyl alcohol Substances 0.000 abstract description 27
- 229920002451 polyvinyl alcohol Polymers 0.000 abstract description 27
- 239000000203 mixture Substances 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 13
- 238000003860 storage Methods 0.000 abstract description 6
- 238000009331 sowing Methods 0.000 abstract description 5
- 230000002411 adverse Effects 0.000 abstract description 3
- 239000003337 fertilizer Substances 0.000 abstract description 3
- 238000002156 mixing Methods 0.000 abstract description 3
- 239000008188 pellet Substances 0.000 abstract description 3
- 235000013311 vegetables Nutrition 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 33
- 241000196324 Embryophyta Species 0.000 description 18
- 238000000034 method Methods 0.000 description 16
- 239000007864 aqueous solution Substances 0.000 description 13
- 239000000047 product Substances 0.000 description 13
- 239000000725 suspension Substances 0.000 description 11
- 230000000813 microbial effect Effects 0.000 description 8
- 230000001954 sterilising effect Effects 0.000 description 8
- 238000004659 sterilization and disinfection Methods 0.000 description 8
- 241000220259 Raphanus Species 0.000 description 7
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 239000008187 granular material Substances 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 5
- 241000233866 Fungi Species 0.000 description 4
- 239000000654 additive Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 230000003042 antagnostic effect Effects 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000005469 granulation Methods 0.000 description 3
- 230000003179 granulation Effects 0.000 description 3
- 239000005416 organic matter Substances 0.000 description 3
- 229960001225 rifampicin Drugs 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000589173 Bradyrhizobium Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 241000607720 Serratia Species 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 238000007664 blowing Methods 0.000 description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 2
- 239000004327 boric acid Substances 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000002361 compost Substances 0.000 description 2
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 235000010958 polyglycerol polyricinoleate Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 235000010413 sodium alginate Nutrition 0.000 description 2
- 239000000661 sodium alginate Substances 0.000 description 2
- 229940005550 sodium alginate Drugs 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 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 1
- 241000589151 Azotobacter Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000589174 Bradyrhizobium japonicum Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241000896533 Gliocladium Species 0.000 description 1
- 229910004835 Na2B4O7 Inorganic materials 0.000 description 1
- 241000244206 Nematoda Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241001668579 Pasteuria Species 0.000 description 1
- 240000001857 Phyllostachys elegans Species 0.000 description 1
- 241000589157 Rhizobiales Species 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 241000082085 Verticillium <Phyllachorales> Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000002154 agricultural waste Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000002734 clay mineral Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000003958 fumigation Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 238000009335 monocropping Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000011368 organic material Substances 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000007678 pseudomonas agar f Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000010801 sewage sludge Substances 0.000 description 1
- 239000003516 soil conditioner Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Soil Conditioners And Soil-Stabilizing Materials (AREA)
Abstract
Description
【0001】0001
【産業上の利用分野】本発明は植物栽培時において、空
中窒素固形菌、拮抗菌(連作障害の主原因である土壌病
害菌や植物寄生性線虫に対して拮抗作用を有する微生物
)、病害抵抗性誘導微生物、PGPR(Plant G
rowth Promoting Rhizobact
eria、植物成長促進性根圏細菌)等の有用な機能を
有する微生物を、植物の成育圏内に持続的に効率よく供
給することが可能な微生物接種用資材に関する。[Industrial Application Field] The present invention is applicable to the use of airborne nitrogen solid bacteria, antagonistic antibacterial bacteria (microorganisms that have an antagonistic effect on soil pathogens and plant parasitic nematodes, which are the main causes of continuous cropping failure), and diseases during plant cultivation. Resistance-inducing microorganism, PGPR (Plant G
rowth Promoting Rhizobact
The present invention relates to a material for inoculating microorganisms that can sustainably and efficiently supply microorganisms having useful functions such as P. elegans, plant growth-promoting rhizobacteria) into the growing range of plants.
【0002】0002
【従来の技術】作物生産において、土壌中の微生物が関
与する役割は大きく、ある種の有用な微生物活性を人為
的に高めることができれば、作物の高位生産を可能にす
る。これらの有用な機能を有する微生物を、植物の成育
圏内に人為的に導入する試みがなされている。BACKGROUND OF THE INVENTION Microorganisms in soil play a major role in crop production, and if the activity of certain useful microorganisms can be artificially increased, high-level production of crops will be possible. Attempts have been made to artificially introduce microorganisms having these useful functions into the growing range of plants.
【0003】微生物の導入法としては次の2種に大別さ
れる。
(1)それぞれが有用な機能を有する微生物をバランス
よく共存させたものを、予め養分となる有機物上で繁殖
させておくか、またはそれらの微生物群と養分となる有
機物を配合したものであり、土壌中に施用することによ
り土壌の微生物相の改良を通じ作物の収量・品質の向上
を図るもので、堆肥または土壌改良材といわれるものを
使用する。
(2)根粒菌、VA菌根菌、拮抗菌、PGPR等の特定
の微生物を植物の成育圏内に導入し、その作用を発現さ
せる微生物資材を使用する方法である。[0003] Methods for introducing microorganisms are broadly classified into the following two types. (1) A well-balanced coexistence of microorganisms, each of which has a useful function, is grown in advance on organic matter that serves as nutrients, or a group of microorganisms and organic matter that serves as nutrients are blended, By applying it to the soil, it aims to improve the yield and quality of crops by improving the microflora of the soil, and uses something called compost or soil conditioner. (2) This is a method of introducing specific microorganisms such as rhizobia, VA mycorrhizal fungi, antagonistic antibacterial fungi, and PGPR into the growing range of plants and using microbial materials to express their effects.
【0004】(1)に属する堆肥や土壌改良材に関する
特許については例えば特開昭60−262886号公報
や特開昭61−209981号公報に開示されている。
これらの資材の欠点は製品の大部分の重量を有機物が占
めることにある。原料の有機物として、稲藁、家畜の糞
尿等の農業廃棄物を用いる場合には農業構造の変化から
原料の入手が難しくなっている。また、比較的供給が安
定している食品工業産廃物(醗酵粕、有機成分抽出残渣
等)や下水汚泥等を有機物原料として使用することも可
能であるが、これらは一般に水分含有量が大きく、しば
しば原料供給地と消費地が離れており、乾燥コストや輸
送コストの面で原料として用いるのに不利がある。[0004] Patents relating to compost and soil improvement materials belonging to (1) are disclosed in, for example, Japanese Patent Laid-Open No. 60-262886 and Japanese Patent Laid-Open No. 61-209981. The disadvantage of these materials is that organic matter accounts for most of the weight of the product. When agricultural waste such as rice straw or livestock manure is used as an organic material, it is becoming difficult to obtain the raw material due to changes in the agricultural structure. It is also possible to use food industry waste (fermentation lees, organic component extraction residue, etc.) and sewage sludge, which are in relatively stable supply, as organic raw materials, but these generally have a high water content and The raw material supply area and the consumption area are often far apart, making it disadvantageous to use it as a raw material in terms of drying costs and transportation costs.
【0005】更に製造過程の問題点として、これらの有
機物と微生物を混合して醗酵させて製造する場合には、
長い製造期間を要し、各種微生物が共存しているため長
期間の保存により微生物相の遷移が起り、場合によって
は効力の低下も生じ、安定した品質の商品を供給するこ
とが困難であること等の問題点がある。中でも、最大の
問題は、仮に上記の問題が全て解決できたとしてもスク
リーニングによって得た最も効果の高い菌株を、他の微
生物群に対して優先的に増殖させることが困難な点にあ
る。[0005] Furthermore, there are problems in the production process when these organic substances and microorganisms are mixed and fermented.
It takes a long manufacturing period, and as various microorganisms coexist, the microbial flora may undergo a transition due to long-term storage, and in some cases, the efficacy may decrease, making it difficult to supply products with stable quality. There are other problems. Among them, the biggest problem is that even if all of the above problems could be solved, it would be difficult to preferentially propagate the most effective bacterial strain obtained through screening over other microbial groups.
【0006】これらの欠点を補うものとして、微生物資
材を使用し、特定の有用微生物を直接接種する方法が提
案されている。その例として根瘤菌を予め種子に封入し
ておく方法、或いは播種時に種子に粉衣する方法が開発
されている。これらの方法では仮に当該微生物が根瘤菌
のように植物の根と親和性があり、且つ運動性を有する
ものであったとしても、根の成長速度が圧倒的に大きい
ため、根の成長に伴って当該微生物が定着することは困
難であり、定着は種子周辺の根域に限定されるという欠
点がった。[0006] In order to compensate for these drawbacks, a method has been proposed in which microbial materials are used and specific useful microorganisms are directly inoculated. For example, a method has been developed in which the root-knot fungus is encapsulated in seeds in advance, or a method in which the seeds are coated with powder at the time of sowing. In these methods, even if the microorganism in question has an affinity for plant roots and is motile, such as root-knot fungi, the growth rate of the roots is overwhelmingly high, so the microorganism is It is difficult for the microorganisms to colonize the seeds, and colonization is limited to the root zone around the seeds.
【0007】この欠点は根瘤菌施用の場合に限定される
ものではなく、例えば植物の根に定着できない微生物を
用いた場合には更に決定的となる。すなわち、施用した
微生物の生存がその土壌に土着する微生物との競合によ
り阻害されるため、効果の持続期間は施用直後のわずか
な期間に限定されてしまう。[0007] This drawback is not limited to the application of root-knot bacteria, but becomes even more decisive when, for example, microorganisms that cannot colonize the roots of plants are used. That is, since the survival of the applied microorganisms is inhibited by competition with microorganisms indigenous to the soil, the duration of the effect is limited to a short period immediately after application.
【0008】この欠点を解決するため、供試する微生物
を固定化し、微生物の活性を植物の成育期間に応じて維
持させ、成育期間に肥料と同様の方法で施用して、活性
な微生物を導入する方法が有効であると考えられている
。上記発想に基づく技術として特開昭62−23400
5号公報に開示さている農林水産業用微生物製剤等があ
り、固定化担体として、アルギン酸ナトリウム、ポリビ
ニルアルコールを用いる方法が提示されている。上記技
術のうち、アルギン酸ナトリウムのゲル化物については
強度の点で問題がある。また、ポリビニルアルコール固
定化物調製法については凍結乾燥、ホウ酸等によるゲル
化法が開示されている。しかしながら、これらの方法は
生菌存在下で急激な温度変化や乾燥の工程或いはホウ酸
等微生物に有害な物質と接触する工程を含み、これらの
条件に弱い微生物の生存性に悪影響を与え、この方法の
現実の実施はかなり制限されるものと考えられる。[0008] In order to solve this drawback, the microorganisms to be tested are immobilized, the activity of the microorganisms is maintained according to the growth period of the plant, and the active microorganisms are introduced by applying it in the same manner as fertilizer during the growth period. This method is considered to be effective. As a technology based on the above idea, Japanese Patent Application Publication No. 62-23400
There are microbial preparations for agriculture, forestry and fisheries industries disclosed in Publication No. 5, and a method using sodium alginate and polyvinyl alcohol as immobilization carriers is proposed. Among the above techniques, the gelled product of sodium alginate has a problem in terms of strength. In addition, freeze-drying, gelation using boric acid, etc. are disclosed as methods for preparing polyvinyl alcohol immobilized products. However, these methods involve rapid temperature changes in the presence of viable bacteria, drying steps, or contact with substances harmful to microorganisms such as boric acid, which adversely affect the survival of microorganisms that are sensitive to these conditions. It is believed that the practical implementation of the method is quite limited.
【0009】[0009]
【発明が解決しようとする課題】本発明者らは、ポリビ
ニルアルコール(以下、PVAとする)と微生物のみの
組合わせでは、製品中の生菌数を安定させることが難し
く実用性に問題があり、土壌と微生物のみで造粒物を製
造した場合には造粒物の強度が不十分で、保管輸送の点
で問題があるという実情に鑑み、製造取扱いが容易で、
保存期間、施用後を通じて生菌が安定して保存され、生
菌数が減少するおそれがなく、播種前に肥料と同時に施
用しても同等の効果を有する微生物接種用資材を提供す
るため鋭意研究したものである。[Problems to be Solved by the Invention] The present inventors found that the combination of only polyvinyl alcohol (hereinafter referred to as PVA) and microorganisms has difficulty in stabilizing the number of viable bacteria in the product and has problems in practicality. In view of the fact that when granules are manufactured using only soil and microorganisms, the strength of the granules is insufficient and there are problems with storage and transportation.
We are conducting extensive research to provide a microbial inoculation material that maintains stable viable bacteria throughout the storage period and after application, has no risk of decreasing the number of viable bacteria, and has the same effect when applied at the same time as fertilizer before sowing. This is what I did.
【0010】0010
【課題解決の手段】本発明は上記課題を解決することを
目的とし、その構成は、滅菌した土壌に水溶性高分子物
質、好ましくはPVAを添加混合したことを特徴とし、
更に、この微生物接種用資材に植物に有用な微生物を配
合して造粒し、好ましくは、ペレット状に造粒したこと
を特徴とする。[Means for Solving the Problems] The present invention aims to solve the above-mentioned problems, and is characterized in that a water-soluble polymeric substance, preferably PVA, is added to and mixed with sterilized soil.
Furthermore, it is characterized in that microorganisms useful for plants are blended into this microorganism inoculation material and granulated, preferably into pellets.
【0011】本発明において使用する土壌は特に限定は
ないが、水田、畑土壌、山野の土壌など肥沃な土壌であ
ることが好ましい。入手できる土壌の性質に応じて炭酸
カルシウム、水酸化カルシウムなどの薬剤を配合してp
H調整をすることもできる。或いはベントナイト、モン
ロリロナイト等の粘土鉱物類を添加し、土壌の物理、化
学的性質を改善することも可能である。また、必要があ
れば、各種有機物を添加し、微生物の生存安定性を増大
させることも可能である。更に、その他の配合物を添加
することも可能である。[0011] The soil used in the present invention is not particularly limited, but it is preferably fertile soil such as paddy field soil, field soil, and mountain soil. Depending on the nature of the available soil, chemicals such as calcium carbonate and calcium hydroxide are mixed into
H adjustment can also be done. Alternatively, it is also possible to improve the physical and chemical properties of the soil by adding clay minerals such as bentonite and montrollilonite. Furthermore, if necessary, various organic substances can be added to increase the survival stability of microorganisms. Furthermore, it is also possible to add other formulations.
【0012】更に、土壌を50〜100タイラーメッシ
ュ程度の篩を通過させたものはその後の造粒が円滑に行
われる。このようにして調製した土壌または土壌と各種
添加物との混合物は滅菌して用いることを要する。滅菌
は通常工業的に行われる高圧蒸気滅菌、エチレンオキサ
イドガス滅菌、放射線滅菌、紫外線滅菌、薬剤滅菌、燻
蒸滅菌等の方法があるが、高圧蒸気滅菌が特に好ましい
。高圧蒸気滅菌の条件は105〜135℃で5〜60分
、好ましくは115〜125℃で15〜30分である。Furthermore, if the soil is passed through a sieve of about 50 to 100 Tyler meshes, subsequent granulation can be carried out smoothly. The soil or the mixture of soil and various additives prepared in this manner must be sterilized before use. Sterilization can be carried out by conventional industrial methods such as high-pressure steam sterilization, ethylene oxide gas sterilization, radiation sterilization, ultraviolet sterilization, chemical sterilization, fumigation sterilization, etc., and high-pressure steam sterilization is particularly preferred. The conditions for autoclaving are 105-135°C for 5-60 minutes, preferably 115-125°C for 15-30 minutes.
【0013】この操作を省略すると、土壌に土着の微生
物と、接種する微生物との間に競合が生じ、接種する微
生物の生存性を著しく損なうことになる。滅菌した土壌
或いは各種添加物と配合した土壌は乾燥しておく。乾燥
条件は室温ないし105℃程度で放置するか、送風下に
乾燥する。好ましくは60〜90℃で5〜24時間であ
る。[0013] If this operation is omitted, competition will occur between the microorganisms indigenous to the soil and the microorganisms to be inoculated, which will significantly impair the survival of the microorganisms to be inoculated. Sterilized soil or soil mixed with various additives should be kept dry. The drying conditions are either left at room temperature to about 105°C, or dried under ventilation. Preferably, the temperature is 60 to 90°C for 5 to 24 hours.
【0014】本発明に用いるPVAは水溶液としても固
形のままでも使用できる。通常PVAを水に溶解したと
きのpHは酸性側であるが目的とする微生物の至適pH
に調整して用いる。水溶液として使用する場合には粘度
1〜300cps程度のものが好適である。粘度1cp
s未満では、でき上がった造粒物の強度が不十分であり
、粘度300cpsを越えると土壌または土壌と添加物
との混合時の操作性が悪く不均一となりやすく、強いて
混合しても微生物の生存性を損なう。PVAの使用量は
接種する土壌または土壌と各種添加物との混合物に対し
、PVA原体に換算して0.5〜30重量部、好ましく
は1〜10重量部である。PVAを固形のまま用いる場
合は、常温での溶解性の点から部分けん化物でかつ微粉
状であることが好ましい。[0014] PVA used in the present invention can be used either as an aqueous solution or as a solid. Normally, the pH when PVA is dissolved in water is on the acidic side, but the optimum pH for the target microorganism is
Adjust and use. When used as an aqueous solution, one with a viscosity of about 1 to 300 cps is suitable. Viscosity 1cp
If the viscosity is less than 300 cps, the strength of the finished granules will be insufficient, and if the viscosity exceeds 300 cps, the operability during mixing of soil or soil and additives will be poor and unevenness will likely occur, and even if forced to mix, microorganisms will not survive. detract from sexuality. The amount of PVA used is 0.5 to 30 parts by weight, preferably 1 to 10 parts by weight, in terms of PVA base material, based on the soil to be inoculated or the mixture of soil and various additives. When PVA is used in solid form, it is preferably partially saponified and in the form of fine powder from the viewpoint of solubility at room temperature.
【0015】本発明に用いる微生物としては各種の微生
物が使用されるが、中でも下記のものが特に好適である
。例えば、リゾビウム属菌(Rhizobium 属菌
)、ブラディリゾビウム属菌(Bradyrhizob
ium属菌)、アゾトバクター属菌 (Azotoba
cter 属菌) 、トリコデルマ属菌 (Trich
oderma 属菌) 、グリオクラジウム属菌(Gl
iocladium 属菌) 、フザリウム属菌(Fu
sarium属菌) 、バーティシリウム属菌 (V
erticillium 属菌) 、エンテロバクター
属菌 (Enterobacter属菌) 、バシルス
属菌 (Bacillus属菌) 、シュードモナス属
菌(Pseudomonas 属菌)、セラチア属菌
(Serratia属菌) 、パステウリア属菌(Pa
steuria 属菌)、ストレプトマイセス属菌(S
treptmyces属菌)等の微生物である。すなわ
ち、本発明では各種微生物を安定して生存させることが
可能である。これら微生物は常法により液体培養または
固体培養を行い、菌体または胞子等の形態で使用する。[0015] Various types of microorganisms can be used in the present invention, and among them, the following are particularly preferred. For example, Rhizobium spp., Bradyrhizobium spp.
ium), Azotobacter (Azotoba genus)
cter genus), Trichoderma genus (Trich
bacteria of the genus Oderma), bacteria of the genus Gliocladium (Gl
iocladium genus), Fusarium genus (Fu
Sarium spp.), Verticillium spp. (V
erticillium), Enterobacter, Bacillus, Pseudomonas, Serratia
(Serratia genus), Pasteuria genus (Pa
steuria spp.), Streptomyces spp. (S
microorganisms such as treptmyces (genus treptmyces). That is, the present invention allows various microorganisms to survive stably. These microorganisms are cultured in liquid or solid form by conventional methods and used in the form of cells, spores, or the like.
【0016】本発明においては土壌とPVAからなる微
生物接種用資材に微生物を配合して成形を行うが、工業
的には例えば下記の方法が使用される。
(1)培養した微生物をPVA水溶液に分散後、パン型
造粒装置を用い土壌または土壌を主体とする配合物に吹
きつけながら造粒物を製造する。必要に応じ微生物に損
傷を与えない温度で乾燥する。In the present invention, microorganisms are blended into a microorganism inoculation material consisting of soil and PVA and molded. For example, the following method is used industrially. (1) After dispersing the cultured microorganisms in a PVA aqueous solution, granules are produced by spraying onto soil or a soil-based composition using a pan-shaped granulator. If necessary, dry at a temperature that does not damage microorganisms.
【0017】(2)培養した微生物をPVA水溶液に分
散後、混練機などを用いて土壌または土壌を主体とする
配合物と混合し、その後、押出成形機等を用いて任意の
形状の押出成形物を製造する。必要に応じ微生物に損傷
を与えない温度で好ましい硬度が得られる迄乾燥する。(2) After dispersing the cultured microorganisms in a PVA aqueous solution, they are mixed with soil or a soil-based mixture using a kneader or the like, and then extruded into an arbitrary shape using an extruder or the like. Manufacture things. If necessary, dry at a temperature that does not damage microorganisms until the desired hardness is achieved.
【0018】(3)培養した微生物をPVA水溶液に分
散後、混練機などを用いて土壌または土壌を主体とする
配合物と混合し、微生物に損傷を与えない温度で好まし
い硬度が得られる迄乾燥後粉砕する。(3) After dispersing the cultured microorganisms in a PVA aqueous solution, they are mixed with soil or a soil-based mixture using a kneader or the like, and dried at a temperature that does not damage the microorganisms until a desired hardness is obtained. Then grind.
【0019】(4)微粉状の部分けん化PVAと土壌ま
たは土壌を主体とする配合物を混合し、パン型造粒装置
を用い培養した微生物の懸濁液を吹きつけながら造粒物
を製造する。(4) Partially saponified PVA in the form of fine powder is mixed with soil or a soil-based composition, and a granulated product is produced by spraying a suspension of cultured microorganisms using a pan-shaped granulator. .
【0020】(5)微粉状の部分けん化PVAと土壌ま
たは土壌を主体とする配合物を混合し、培養した微生物
の懸濁液を注入混合し、(2)及び(3)に方法により
成形物を製造する。(5) Mix partially saponified PVA in the form of fine powder and soil or a soil-based mixture, inject and mix a suspension of cultured microorganisms, and form a molded product according to the methods of (2) and (3). Manufacture.
【0021】(6)PVA溶液に土壌または土壌を主体
とする配合物を混合、造粒し、好ましい硬度またはこれ
よりやや硬く乾燥した後、培養した微生物の懸濁液を吹
きつけながら造粒物を製造する。(6) Mix soil or a soil-based mixture with the PVA solution, granulate it, dry to a desired hardness or slightly harder, and then spray the granulated product with a suspension of cultured microorganisms. Manufacture.
【0022】[0022]
【作用】PVAと微生物のみの組合わせでは前述の通り
、製品中の生菌数を安定させることが難しく実用性に問
題があり、土壌と微生物のみで造粒物を製造した場合に
は造粒物の強度が不十分で、保管輸送及び微生物の土壌
中における生存性の点で問題があった。本発明は土壌の
有する適度の栄養性と造粒性に着眼し、更にPVAを使
用することにより好ましい造粒性と生菌安定性を有する
微生物接種用資材を提供することに成功したものである
。この資材を利用して、植物に有用な機能を有する微生
物を資材に固定し、播種前または播種後の土壌に混合す
ることにより植物の成長に合わせて充分な量の有用微生
物を生存させることが可能になり、これらを植物の根に
連続的に供給することができる。[Effect] As mentioned above, with the combination of only PVA and microorganisms, it is difficult to stabilize the number of viable bacteria in the product and there are problems with practicality. The strength of the product was insufficient, and there were problems in terms of storage, transportation, and survival of microorganisms in the soil. The present invention focuses on the appropriate nutritional properties and granulation properties of soil, and has succeeded in providing a material for microbial inoculation that has favorable granulation properties and viable bacteria stability by using PVA. . By using this material, microorganisms that have useful functions for plants are immobilized on the material, and by mixing it with the soil before or after sowing, it is possible to keep a sufficient amount of useful microorganisms alive as the plants grow. This makes it possible to continuously supply these to the roots of plants.
【0023】[0023]
【発明の効果】本発明微生物接種用資材は、保存中、土
壌中においても生菌数が安定しているため、植物栽培前
の土壌に事前に施用しておくことが可能であり、作業が
省力化され、種々の有用菌の植物栽培への応用範囲が拡
大する。また、土壌に大量に供給しても土壌中の菌との
間に調和が得られるため、生態系に悪影響を与えるおそ
れがない。更に、育苗土に本発明品を混合して使用し、
育苗土ごと本圃に移植しても製品中の生菌数が安定性が
優れているため、以後の生菌数も安定して、植物に有用
な効果をもたらす。[Effect of the invention] The microorganism inoculation material of the present invention has a stable number of viable bacteria even during storage and in the soil, so it can be applied in advance to the soil before cultivating plants, making the work easier. This saves labor and expands the scope of application of various useful bacteria to plant cultivation. In addition, even if large amounts are supplied to the soil, there is no risk of adverse effects on the ecosystem because harmony is achieved with the bacteria in the soil. Furthermore, the product of the present invention is mixed and used in seedling raising soil,
Since the number of viable bacteria in the product is highly stable even when transplanted to the main field along with the seedling soil, the number of viable bacteria in the future will also be stable, resulting in a beneficial effect on the plants.
【0024】[0024]
【実施例】以下、本発明を実施例、比較例により具体的
に説明する。
実施例1
大根の根に定着することのできる分離株である蛍光性P
seudomonas 属細菌、DK−240「 (R
ifanpicin耐性株)を液体培地で28℃で、好
気的条件下で培養を行い、PVA B−17(電気化
学工業社製)と、電気化学工業株式会社、総合研究所内
土壌を用いて造粒化した。[Examples] The present invention will be specifically explained below using Examples and Comparative Examples. Example 1 Fluorescent P, an isolate capable of colonizing the roots of radish
bacterium of the genus pseudomonas, DK-240 (R
ifanpicin resistant strain) was cultured in a liquid medium at 28°C under aerobic conditions, and granulated using PVA B-17 (manufactured by Denki Kagaku Kogyo Co., Ltd.) and soil within the Research Institute of Denki Kagaku Kogyo Co., Ltd. It became.
【0025】DK−240株は神奈川県三浦市の大根圃
場で栽培した大根の根から分離した細菌である。この細
菌はブイヨン寒天培地で良好な成育を示し、極鞭毛を有
し、運動性があり、グラム陰性の絶対好気性の桿菌で胞
子を作らない。また、カタラーゼ活性が陽性で、O−F
(High−Leifson) テストにより好気的に
酸を生成し、Pseudomonas agar F
(PAF)培地上で28℃、2日間培養すると蛍光性の
色素(フルオレシン)を産生することから蛍光性Pse
udomonas 細菌であると同定した。DK−24
0株から自然変異により生じたRifanpicin
耐性株(Rifanpicin100μg/ml倍で
成育可)を分離し、DK−240「 株と命名した。DK-240 strain is a bacterium isolated from the roots of radish grown in a radish field in Miura City, Kanagawa Prefecture. This bacterium grows well on broth agar, is polar flagellated, motile, Gram-negative, obligately aerobic rod, and does not produce spores. In addition, catalase activity was positive, and O-F
(High-Leifson) test produces acid aerobically and Pseudomonas agar F
When cultured on (PAF) medium at 28°C for 2 days, a fluorescent dye (fluorescin) is produced.
It was identified as a udomonas bacterium. DK-24
Rifanpicin generated by natural mutation from strain 0
A resistant strain (which can grow at 100 μg/ml of Rifanpicin) was isolated and named strain DK-240.
【0026】用いたPVA B−17は温浴中(90
℃以上)で溶解して水溶液とし、冷却後pH6.9〜7
.1に調整した。このPVA水溶液5mlに菌懸濁液1
.2mlを添加して微生物含有PVA水溶液中の濃度が
5%になるように調整した。[0026] The PVA B-17 used was heated in a warm bath (90
℃ or above) to form an aqueous solution, and after cooling, the pH is 6.9 to 7.
.. Adjusted to 1. Add 1 part of the bacterial suspension to 5 ml of this PVA aqueous solution.
.. 2 ml was added to adjust the concentration in the microorganism-containing PVA aqueous solution to 5%.
【0027】この微生物含有PVA水溶液(生菌数10
12個/mlを含む)に土壌10gを添加した後よく混
合し、注射筒を用いて押出して直径1mm、長さ5〜6
mmの円柱状に造粒した。含水率が30〜40%になる
ように室温で送風下に乾燥し、目的とする微生物含有資
材を得た。この資材1g中には2×1010個の生菌が
存在していた。得られた微生物接種用資材をポットによ
る植物栽培試験に供した。[0027] This microorganism-containing PVA aqueous solution (viable number of bacteria: 10
After adding 10g of soil to the mixture (containing 12 particles/ml), mix well and extrude using a syringe to obtain a mixture with a diameter of 1mm and a length of 5-6.
The pellets were granulated into a cylindrical shape of mm. The material was dried at room temperature under blowing air so that the moisture content was 30 to 40%, and the desired microorganism-containing material was obtained. There were 2×1010 live bacteria in 1 g of this material. The obtained microbial inoculation materials were subjected to a plant cultivation test using pots.
【0028】300gの土壌を入れたポットの2箇所に
各造粒物を1カ所当り0.15g施用した。施用した日
及び5週間後に大根の種子を造粒物施用位置に播種し、
温室内で栽培した。各々播種後6週間目に大根の根部を
採集し、滅菌水中で付着する土壌を除去した後、滅菌水
中でホモジナイズし、この大根の根部懸濁液をrifa
mpicin100μg/ml含有PAF培地上に接種
することにより、その根面から供試菌を回収し、大根の
根部重量当りの生菌数を算出した。結果を表1に示した
。[0028] Each granulated product was applied at two locations in a pot containing 300 g of soil at a rate of 0.15 g per location. On the day of application and 5 weeks later, radish seeds are sown at the granule application position,
Grown in a greenhouse. The roots of each radish were collected 6 weeks after sowing, and after removing the attached soil in sterile water, they were homogenized in sterile water, and the root suspension of this radish was rifated.
Test bacteria were collected from the root surface by inoculating them onto a PAF medium containing 100 μg/ml of mpicin, and the number of viable bacteria per root weight of the radish was calculated. The results are shown in Table 1.
【表1】[Table 1]
【0029】比較例1
DK−240「 株をNutrient broth
( Difco )+1%グルコース培地で培養後10
0mlを分取し、遠心分離を行い上清を廃棄した。残っ
た湿潤菌体にPVAB−17の10%水溶液100ml
を添加混合し、生菌109 個を含むPVA水溶液を調
製した。調製した生菌懸濁液は、0.7 m/m 注
射針を装着したシリンダからNa2 B4 O7 飽和
水溶液(pH7.0 )中に滴下し、球状の生菌含有P
VAゲル化担体を調製した。調製した直後の担体中には
2×108 個/粒の生菌が含まれていた。この担体を
ポットにおける植物栽培実験に供試した。植穴当り施用
量は初期の生菌量をそろえるため植穴当り15粒/植穴
とした他は実施例1に準じて行った。その結果を表1に
併記した。Comparative Example 1 DK-240
(Difco)+10 after culturing in 1% glucose medium
0 ml was collected, centrifuged, and the supernatant was discarded. Add 100 ml of a 10% aqueous solution of PVAB-17 to the remaining moist bacterial cells.
were added and mixed to prepare a PVA aqueous solution containing 109 live bacteria. The prepared live bacteria suspension was dropped into a Na2B4O7 saturated aqueous solution (pH 7.0) from a cylinder equipped with a 0.7 m/m injection needle, and spherical live bacteria-containing P
A VA gelling carrier was prepared. The carrier immediately after preparation contained 2 x 108 live bacteria/grain. This carrier was used in a plant cultivation experiment in a pot. The application amount per planting hole was 15 seeds/planting hole in order to equalize the initial amount of viable bacteria, but the procedure was as in Example 1. The results are also listed in Table 1.
【0030】実施例2
Bradyrhizobium japonicum
IFO 15001から突然変異処理によりrifam
picin耐性変異株(100μg rifampi
cin/ml培地で成育可)を取得した。上記変異株は
rifampicin100μg/mlを含む酵母エキ
ス・マンニット寒天培地上で30℃、1週間培養の後集
菌した。集菌した変異株はpH7.0に調整したPVA
B−17の7%水溶液に撹拌懸濁し生菌2×101
1個/mlの懸濁液を調製した。滅菌した土壌10gに
懸濁液8mlを添加混合し、注射筒より押出し成形し、
送風下で水分25%まで乾燥した。担体中には1×10
10個/gの生菌が存在していた。Example 2 Bradyrhizobium japonicum
rifam from IFO 15001 by mutation treatment
picin-resistant mutant strain (100 μg rifampi
cin/ml medium) was obtained. The above mutant strain was cultured on a yeast extract/mannite agar medium containing 100 μg/ml of rifampicin at 30° C. for one week and then harvested. The collected mutant strain was treated with PVA adjusted to pH 7.0.
Stir and suspend live bacteria in a 7% aqueous solution of B-17 and add 2 x 101
A suspension of 1 cell/ml was prepared. Add and mix 8 ml of the suspension to 10 g of sterilized soil, extrude through a syringe,
It was dried under blowing air to a moisture content of 25%. 1 x 10 in the carrier
There were 10 live bacteria/g.
【0031】上記担体1gを畑土壌100gにV型混合
機を用いて均一に混合した。担体を含む畑土壌はポット
に充填し、最大容水量の60%の水分を保つように水分
を補給しながら恒温庫中で28℃でインキュベートを行
った。経時的に土壌全量を回収し、土壌懸濁液を調製し
、rifampicin含有酵母エキス・マンニット寒
天培地への希釈平板法により土壌供試変異株数を測定し
、その結果を表2に示した。1 g of the above carrier was uniformly mixed with 100 g of field soil using a V-type mixer. The field soil containing the carrier was filled into a pot, and incubated at 28° C. in a thermostatic chamber while replenishing water to maintain 60% of the maximum water capacity. The total amount of soil was collected over time, a soil suspension was prepared, and the number of soil test mutants was measured by the dilution plate method on a rifampicin-containing yeast extract/mannite agar medium. The results are shown in Table 2.
【表2】[Table 2]
【0032】比較例2
実施例2と同様に、Bradyrhizobium j
aponicum IFO 15001の突然変異株を
培養した。変異株は無菌水に懸濁し、生菌数5×108
個/mlの生菌懸濁液を得た。この懸濁液20mlを
スポイトを用い土壌100gに均一に滴下後更にV型混
合機にて充分に撹拌混合した。実施例2に準じて土壌中
の生菌数を測定し、表2に併記した。Comparative Example 2 Similar to Example 2, Bradyrhizobium j
A mutant strain of Aponicum IFO 15001 was cultured. The mutant strain was suspended in sterile water and the number of viable bacteria was 5 x 108.
A suspension of viable cells/ml was obtained. 20 ml of this suspension was uniformly dropped onto 100 g of soil using a dropper, and then sufficiently stirred and mixed using a V-type mixer. The number of viable bacteria in the soil was measured according to Example 2 and is also listed in Table 2.
【0033】表1及び表2から明らかな通り、本発明微
生物接種用資材は土壌中において長期にわたり特定の微
生物を安定して生存させる効果を有することが判明した
。As is clear from Tables 1 and 2, it was found that the microorganism inoculation material of the present invention has the effect of stably allowing specific microorganisms to survive in the soil for a long period of time.
Claims (2)
加混合したことを特徴とする微生物接種用資材。1. A material for inoculating microorganisms, characterized in that a water-soluble polymer substance is added and mixed with sterilized soil.
分子物質とを含有したことを特徴とする微生物接種用資
材。2. A material for inoculating microorganisms, comprising microorganisms, sterilized soil, and a water-soluble polymer substance.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3156006A JP2972390B2 (en) | 1991-05-31 | 1991-05-31 | Materials for microbial inoculation |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3156006A JP2972390B2 (en) | 1991-05-31 | 1991-05-31 | Materials for microbial inoculation |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH04353593A true JPH04353593A (en) | 1992-12-08 |
JP2972390B2 JP2972390B2 (en) | 1999-11-08 |
Family
ID=15618271
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP3156006A Expired - Fee Related JP2972390B2 (en) | 1991-05-31 | 1991-05-31 | Materials for microbial inoculation |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP2972390B2 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0656614A (en) * | 1992-07-31 | 1994-03-01 | Central Glass Co Ltd | Agricultural chemical of microorganism |
-
1991
- 1991-05-31 JP JP3156006A patent/JP2972390B2/en not_active Expired - Fee Related
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0656614A (en) * | 1992-07-31 | 1994-03-01 | Central Glass Co Ltd | Agricultural chemical of microorganism |
Also Published As
Publication number | Publication date |
---|---|
JP2972390B2 (en) | 1999-11-08 |
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