JP3665845B2 - Artificial culture method of Kyushu insecticide - Google Patents
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Description
【０００１】
【発明の属する技術分野】
本発明は一種の微生物菌類を人工的に培養する方法に関するものであり、特に九州虫草菌と名づけられた中国で発見された新種を培養し、その子実体を実際に応用する方法に関する。
【０００２】
【従来の技術】
九州虫草菌は、一般的に利用されている菌類の一種である。野生の九州虫草菌は昔から大切にされてきた中国の薬の材料であって、その多くの薬理作用や栄養効果のために消費者の間で人気のあるものとなっている。しかし、九州虫草菌の生育環境が極めて特殊であるという理由から産出量が少なく、消費者の需要に応えられる状況にはなっていない。
【０００３】
現在多くの研究者達がその人工培養方法について研究し、資源として不足している状態を解決しようと試みている。九州虫草菌の人工培養方法に関しては次に示す文献が開示されており、「冬中夏草の菌糸体の工業生産方法」（中国特許第８５１０７９９１号）及び「冬中夏草の菌株８１４の選択及び発酵技術」（中国特許第８５１０２２３１号）には、菌糸体が念入りな培養及び発酵処理の後の純粋な菌株によって得られると報告されている。これまで九州虫草菌は発生の過程が完了した状態で発見されるため、人工的培養を行なうことができなかった。
【０００４】
「蚕蛹虫草の人工培養方法」（中国特許第８７１０６９８７号）では、ある程度繁殖している菌糸を虫に注入して虫草をバーティシリウムで培養させるために固体培養媒体を使用して、その虫に自然に感染させている。子実体が形成された後は、子実体と虫とを共に使用して薬を製造している。しかし、培養時間は長いが、子実体は虫の体の１／５しか成長していない。
【０００５】
「ブラジル虫草菌の人工培養方法」（中国特許第９０１０９３８０．７号）では接種によって虫と菌類の結合体を得ることが報告されているが、ここに述べられている九州虫草菌の利用例は、純粋な子実体を用いたものではない。「天津薬理学」（第１０巻（１）、１９９８年）の報告によると、虫草菌をマウスに投与した場合、マウスの脾臓の重さが明らかに増加するため、虫草菌の作用部分が虫の体ではなく菌糸束の中にあることが示されている。
【０００６】
「冬中夏草胞子と有性的段階の完成技術」（中国特許第８８１０３４５３号）には、冬中夏草の胞子は九州虫草菌の成長したものであり、菌の種子は数ヶ月後に胞子を生成することができると記録されている。研究者らは更に、１０℃での処理を行い、１８℃で培養した成長胞子を使用し寄生主であるコウモリの幼体に接種させた。しかし生成能力が失われた突然変異効率の低い胞子があるので、接種後では寄生の効率も低かった。そのため、この方法で広く使用されている子実体効果を形成するのは困難である。
【０００７】
本願の発明者はかつて、「北冬中夏草の優れた菌株の選択方法」（中国特許第９３１０５０５６．１号）、「蛹虫草の優れた菌株の選択及び高い生産性での培養方法」（第９４１１００６１８号）及び「高い生産性の蛹虫草の培養方法」（中国特許第９７１０５８５３１０号）等の特許文書において、ノイムシダケ属菌についての人工培養方法を報告している。この方法では、マウスが固体培養媒体の主成分であり、この方法をとることにより蛹虫草菌を培養することに成功した。
【０００８】
しかし、培地物質は単純であるため、栄養が不充分であり、子実体を十分に成長させることができなかった。このことにより、菌株の退化及び低下する薬理学的活動が明らかになった。
【０００９】
更に、「菌類系統」（第１９巻（２）２９６頁、２０００年）では、中国で新しく発見されたきのこ類の菌株、九州虫草菌（Ｃｏｒｄｙｃｅｐｓ Ｋｙｕｓｈｕｅｎｓｉｓ Ｙ．ｋｏｂａｙａｓｉ）は豆天蛾（Ｃｌａｎｉｓ Ｂｉｌｉｎｅａｔａ Ｗａｌｋｅｒ）の幼虫の中でその寄生生活を送ると述べられている。その外形及び特徴は以下の通りである。子実体は幼虫体から白い菌糸束を伸ばして成長する。菌糸束は黄みがかったオレンジで、高さ１．７〜８．６ｃｍ、直径０．２〜０．６ｃｍ、被子器は半内部卵形で３６７〜４９５×２６０〜３１６μｍ、菌糸束は柱状で直径が３．５〜４．８μｍ。二次胞子は短柱状で３．９〜５．４×１．０〜１．２μｍである。しかし、現在のところ、本国又は海外における九州虫草菌の人工培養方法についての報告はない。
【００１０】
【発明が解決しようとする課題】
しかしながら、これまで九州虫草菌は発生の過程が完了した状態で発見されるため、人工的培養を行なうことができなかった。また、培養時間は長いが、子実体が虫の体の１／５しか繁殖していなかったり、生成能力が失われた突然変異効率の低い胞子があるので、接種後では寄生の効率も低かった。また、培地物質が単純であり、体の栄養バランスからみて不充分な組成であるため、子実体を十分に成長させることができなかった。このため上記の全ての方法は有効栄養成分の濃度の低下を引き起こすため、菌株の安定性を保証することができず、蛹虫草菌の子実体を連続的に培養することが困難であった。
【００１１】
【課題を解決するための手段】
本発明は上記の問題点を解決するためになされたものであり、請求項１の発明は九州虫草菌の人工培養方法であり、野生の九州虫草菌を単一の胞子又は単一のコロニーに純化した菌株を液体菌種にして、無菌状態で飼育された豆天蛾（Ｃｌａｎｉｓ Ｂｉｌｉｎｅａｔａ Ｗａｌｋｅｒ）幼虫の凍結乾燥粉末を３〜５重量部、穀物を９７〜９５重量部と、これら材料合計量の１．４倍の栄養水を加えた組成でなる混合培養媒体に植え付けられ、日光と風をあてながら九州虫草菌子実体を生成させるものである。
【００１２】
請求項２の発明は九州虫草菌の人工培養方法であり、請求項１における液体菌種が、前記の純化された菌株を、無菌状態で飼育された３〜５齢の豆天蛾幼虫の凍結乾燥粉末を３〜５重量部、ジャガイモを１０〜１５重量部、、砂糖を１〜２重量部、ビタミンＢ １ を０ ．００１〜０．００２重量部含有し、残部が水で全体を１００重量部としてなる液体培養媒体に培養したものである。
【００１３】
請求項３の発明は九州虫草菌の人工培養方法であり、請求項１における栄養水が、九州虫草菌子実体の精製粉末を１〜１．５重量部、ＫＨ ２ ＰＯ ４ を２〜２．５重量部、Ｋ ２ ＨＰＯ ４ を１〜１．５重量部、ＭｇＳＯ ４ を１〜１．５重量部含有し、残部が水で全体を１０００重量部とした組成である。
【００１４】
【発明の実施の形態】
以下、本発明の実施の形態を説明する。
図１は野生の菌株の形態学的図であり、図２は蛾の幼虫から作られた混合培養媒体で培養された菌株の形態学的図であり、図３は混合培養媒体で培養された菌株の形態学的図である。図１〜図３を参照すると、野生の菌株と培養された菌株は形態学特徴が類似していることが分かる。本発明の培養方法はその他の菌の培養方法とは異なり、主な特徴は以下の通りである。
（１）収集された野生の菌株の組織又は胞子を分ける。
（２）単一の菌のコロニー又は単一の胞子を精製して培養する。
（３）顕微鏡で菌糸を観察して、精製された九州虫草菌の特徴と一致する菌株を分け、これを用いて液体菌種を作る。
（４）無菌状態で飼育された豆天蛾幼虫幼虫の凍結乾燥粉末を穀物と混合し、栄養水を加えた混合培養媒体に、上記液体菌種を植え付ける。混合培養媒体が菌糸でいっぱいになるまで、自然感染させる。適当な状態で、野生の菌株と同じ子実体に成長することが可能である。
【００１５】
ここで液体菌種は、無菌状態で飼育された３〜５齢の蛾幼虫の凍結乾燥粉末を３〜５重量部、ジャガイモを１０〜１５重量部、砂糖を１〜２重量部、ビタミンＢ １ を０．００１〜０．００２重量部含有し、残部が水で全体を１００重量部とした培養液に、純化された菌株が植え付けられたものである。
【００１６】
混合培養媒体は、無菌状態で飼育された３〜５齢の豆天蛾幼虫の凍結乾燥粉末が３〜５重量部、米、トウモロコシ粉末等の穀物が９７〜９５重量部と、これら材料合計量の１．４倍の栄養水を加えた組成である。ここで、栄養水は、精製された九州虫草菌子実体の粉末を１〜１．５重量部、ＫＨ２ＰＯ４を２〜２．５重量部、Ｋ２ＨＰＯ４を１〜１．５重量部、ＭｇＳＯ４を１〜１．５重量部含有し、残部が水で全体を１０００重量部としている。
【００１７】
本願発明の培養方法によれば、九州虫草菌を精製または選択する過程で純粋な九州虫草菌が選択できることだけでなく、人工培養状態で作った九州虫草菌子実体が野生のものと同じであるということが保証できる。また、その他の人工培養方法と比べると、生産周期が短く、菌株が安定し、接種効率が明らかに増加し、生存率と子実体生成率が１００％以上であり、形質転換率が１６８．３％であるという多くの利点があり、連続的に大規模生産できる基盤とすることができる。また、薬の活性を阻害する傾向も防ぐことができる。瀋陽応用生態研究所、ＣＡＳ及び遼寧省中国新薬開発研究所によって、この九州虫草菌は外形が野生のものと類似しているという報告がなされており、その結果を表１に示している。
【００１８】
【表１】
また、得られた九州虫草菌子実体の栄養成分を表２に示した。
【００２０】
【表２】
これらの点から、有効栄養成分の濃度は高いレベルに達していることがわかる。九州虫草菌子実体を薬の材料として使用して健康製品を製造することにより、肺に栄養を与える、腎臓に良い、止血する、痰を分解する、及び健康状態を改善する、弱った肺や腎臓によって起こった病気を治す機能を強めることができる。また、それだけなく腫瘍の正常化（免疫調節機能の改善）、体に良くない物を追い出す（腫瘍の増殖抑制及び生命の延長）、毒性の軽減（医学的治療の造血細胞の機能の減少）のような面でも使用することができ、腫瘍細胞の増殖を抑制し、疲労からの回復、及び体の免疫機能を改善することができる。
【００２２】
九州虫草菌子実体の有効栄養成分は比較的高いレベルなので、少量の材料でも薬理学的機能を損なうことなく、費用を少なくすることができる。このために、上記に説明された方法は科学技術部門において研究される主要な項目として取り扱われ、新薬の第一級のものとして研究されてきた。
【００２３】
この実用化するに、発明による九州虫草菌子実体を乾燥し、１００メッシュの粉末に粉砕してからアルコールを用いて造粒し、６０℃以下で粒子を乾燥させ、Ｃｏ６０で殺菌したカプセルに入れる、あるいは、九州虫草菌子実体にマンニトール、ステアリン酸マグネシウム、サッカリンナトリウム、メントールなどを配合し、アルコールを用いて造粒し、乾燥させて九州虫草菌入りのの錠剤とすることができる。カプセル、錠剤の他にも、食用薬品、健康食品、経口液、飲料及びアルコール飲料の有効成分とすることも可能である。少量の材料で、その機能は顕著なものである。
【００２４】
（実施例１）
収集された野生の九州虫草菌子実体の外側被膜をはがし、消毒済みのナイフで０．２〜０．３ｃｍの組織を切り取り、殺菌された媒体の入っている培養皿に置いて表面接種させた。適温で、７日間培養した後、強い菌糸を取り出して試験管の斜面に置き、さらに７〜１０日間培養して、大まかに選択し菌糸の成長状態に基づいて培養皿に植え付けた。カバーガラスを、植え付けた部分のまわり全てに挿入し、５日後カバーガラス上の菌糸を顕微鏡で観察し、その菌株の特徴に合う菌糸を液体菌種とした。尚、液体菌種は、１０ｍｇの純粋な菌株を、豆天蛾の幼虫の凍結乾燥粉末：３〜５重量部、ジャガイモ：１０〜１５重量部、砂糖：１〜２重量部、ビタミンＢ１：０．００１〜０．００２重量部及び残りが水で全体を１００重量部とした培養液に、植え付けたものである。
【００２５】
（実施例２）
豆天蛾幼虫の凍結乾燥粉末３ｋｇ、ジュズダマ１０ｋｇ、粉砕したトウモロコシ１２ｋｇ、米７５ｋｇを一様に混ぜ、この混合物３０ｇと栄養水４２ｍｌをプラスチックボトルに詰めた。殺菌後、２ｍｌの液体菌種子を植え付け、適温で約２０日間培養した。媒体の表面が下に伸びている菌糸でいっぱいになったら、分散光（約２５００Ｌｕｘ）をあてて、換気した。（膜がボトルの縁に付着している。）１日３〜４回、１回につき２０〜３０分間ドアと窓を開けた。室内の湿度は８５〜９０％であった。２５〜３０日後、子実体が得られた。
【００２６】
（実施例３）
豆天蛾幼虫の凍結乾燥粉末４ｋｇ、ジュズダマ９ｋｇ、粉砕したトウモロコシ１１ｋｇ、米７６ｋｇを一様に混ぜ、この混合物４０ｇと栄養水５６ｍｌをボトルに詰めた。殺菌後、３ｍｌの液体菌種子を植え付けて、光なしの適温状態で約２３日間培養した。媒体の表面が下に伸びている菌糸でいっぱいになったら、分散光を与えた。
【００２７】
（実施例４）
豆天蛾幼虫の粉末５ｋｇ、ジュズダマ８ｋｇ、粉砕したトウモロコシ１０ｋｇ、米７７ｋｇを一様に混ぜ、この混合物５０ｇと栄養水７０ｍｌをボトルに詰めた。殺菌後、４ｍｌの液体菌種子を植え付けて、適温で約２５日間培養させた。媒体の表面が下に伸びている菌糸でいっぱいになったら、分散光をあてた。
【００２８】
【発明の効果】
本発明によれば、野生の九州虫草菌の単一の胞子又はコロニーを精製した菌株から、最終的に子実体が形成されるまで任意の培養状態で自然感染させるので、生産周期が短く、菌株の退化現象が起こらずに安定していて、感染効率が増加し、生存率と子実体生成率が１００％以上であり、形質転換率が１６８．３％である子実体の培養を行うことができ、連続的な大規模生産が可能にすることができる。また、この子実体を薬の材料として使用すると、少量で、効果のある薬剤とすることができ、また健康食品、飲料などの有効成分として用いることができる。
【図面の簡単な説明】
【図１】 野生の菌株の形態学的図である。
【図２】 豆天蛾の幼虫から作られた培養媒体で培養された菌株の形態学的図である。
【図３】 豆天蛾の幼虫から作られた培養媒体で培養された菌株の形態学的図である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method of artificially cultivating a kind of microbial fungi, and more particularly to a method of culturing a new species discovered in China named Kyushu insect fungus and applying its fruiting bodies in practice.
[0002]
[Prior art]
Kyushu insect fungus is a kind of commonly used fungi. Wild Kyushu fungus is a Chinese medicine material that has been treasured since ancient times, and has become popular among consumers due to its many pharmacological and nutritional effects. However, because the growth environment of Kyushu insecticide is extremely special, the production volume is small and it is not in a situation that can meet the demand of consumers.
[0003]
Many researchers are now studying the artificial culture method and trying to solve the shortage of resources. Regarding the artificial culture method of Kyushu insecticide, the following documents are disclosed: “Industrial production method of mycelium of winter summer summer” ( Chinese Patent No. 85107991) and “Selection of strain 814 of winter summer summer grass” And fermentation technology "( Chinese Patent No. 85102231) reports that mycelia are obtained by pure strains after careful culture and fermentation treatment. Until now, Kyushu insecticides have been discovered in a state where the development process has been completed, and thus artificial culture has not been possible.
[0004]
In the method of artificial culture of helminths ( Chinese Patent No. 87106987), a solid culture medium is used to inject worms that have been propagated to some extent and cultivate worms in Verticillium. It is naturally infected. After the fruiting body is formed, drugs are produced using both the fruiting body and insects. However, although the culture time is long, the fruiting body grows only 1/5 of the insect body.
[0005]
In the “Artificial culture method of Brazilian insect fungus” ( Chinese Patent No. 90109380.7), it is reported that a combination of insects and fungi is obtained by inoculation. It is not a pure child entity. According to a report of “Tianjin Pharmacology” (Volume 10 (1), 1998), when protozoan fungus is administered to mice, the weight of the spleen of the mouse clearly increases, so It is shown to be in the mycelium bundle, not the body.
[0006]
According to “Winter Summer Soybean Spore and Completion Technique of Sexual Stage” ( Chinese Patent No. 88103453), winter summer summer spores are grown by Kyushu insect fungus and fungus seeds are spores after several months. Can be generated and recorded. The researchers further treated at 10 ° C. and used the grown spores cultured at 18 ° C. to inoculate larvae of the main parasite bats. However, since there were spores with low mutation efficiency that lost production capacity, parasitic efficiency was low after inoculation. Therefore, it is difficult to form a fruit body effect widely used in this method.
[0007]
The inventor of the present application once said, “Selection method of excellent strains of northern winter midsummer” ( Chinese Patent No. 93105056.1), “Selection of superior strains of helminths and culture method with high productivity” ( No. 941100688) and “a highly productive method for cultivating helminths” ( Chinese Patent No. 9710585310), and the like, have reported an artificial culture method for the genus Noimushida. In this method, the mouse is the main component of the solid culture medium, and by using this method, helminths were successfully cultured.
[0008]
However, since the medium material is simple, the nutrition is insufficient and the fruiting body cannot be grown sufficiently. This revealed a degenerative and reduced pharmacological activity of the strain.
[0009]
Furthermore, in the “Fungus strain” (Vol. 19 (2) 296, 2000), a newly discovered mushroom strain in China, Cordyceps Kyushuensis Y. kobayasi, is a Clarice Bilineta Walker. ) Larvae are said to live their parasitoids. The outline and features are as follows. The fruiting body grows from a larval body by extending a white mycelium bundle. The mycelium bundle is yellowish orange with a height of 1.7 to 8.6 cm, a diameter of 0.2 to 0.6 cm, the angioplasty is a semi-inner egg shape, 367 to 495 × 260 to 316 μm, and the mycelium bundle is columnar. The diameter is 3.5 to 4.8 μm. The secondary spores are short columnar and 3.9 to 5.4 × 1.0 to 1.2 μm. However, at present, there is no report on an artificial culture method of Kyushu insecticide in Japan or overseas.
[0010]
[Problems to be solved by the invention]
However, since Kyushu insecticide has been discovered in a state where the development process has been completed, it has not been possible to perform artificial culture. In addition, although the culture time is long, there are spores with low mutation efficiency that the fruiting bodies have propagated only 1/5 of the body of the worm or lost the production ability, so the parasitic efficiency was also low after inoculation . Further, a simpler medium material, because it is insufficient composition viewed from the nutrient balance of the body, it was not possible to sufficiently grow the fruiting bodies. For this reason, since all the above methods cause a decrease in the concentration of effective nutrients, the stability of the strain cannot be guaranteed, and it has been difficult to continuously culture the fruit bodies of Helminthiamyces.
[0011]
[Means for Solving the Problems]
The present invention has been made to solve the above-mentioned problems, and the invention of claim 1 is an artificial culture method for Kyushu insect fungus, wherein the wild Kyushu fungus is transformed into a single spore or a single colony. 3-5 parts by weight of lyophilized powder and 97-95 parts by weight of cereal larvae (Clanis Bilineta Walker) larvae cultivated in a sterile state with the purified strain as a liquid strain , It is planted in a mixed culture medium having a composition containing 1.4 times nutrient water, and produces Kyushu insect fungus fruit bodies while applying sunlight and wind .
[0012]
The invention of claim 2 is an artificial culture method for Kyushu insecticide, wherein the liquid bacterial species in claim 1 is a freezing of 3-5 year old bean larvae bred in aseptic condition. 3-5 parts by weight of dry powder, 1-2 parts by weight 10 to 15 parts by weight ,, sugar potatoes, vitamin B 1 0. It is cultured in a liquid culture medium containing 001 to 0.002 parts by weight, the balance being water and the whole being 100 parts by weight.
[0013]
The invention of claim 3 is a method for artificial culture of Kyushu insect fungus, wherein the nutrient water in claim 1 is 1 to 1.5 parts by weight of purified powder of Kyushu insect fungus fruiting body and 2 to 2.5 of KH 2 PO 4 . The composition is 1 to 1.5 parts by weight , 1 to 1.5 parts by weight of K 2 HPO 4 and 1 to 1.5 parts by weight of MgSO 4 , with the remainder being water and 1000 parts by weight as a whole.
[0014]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described below.
FIG. 1 is a morphological view of a wild strain, FIG. 2 is a morphological view of a strain cultured in a mixed culture medium made from pupae larvae , and FIG. 3 is cultured in a mixed culture medium. It is a morphological view of a strain. 1-3, it can be seen that wild strains and cultured strains have similar morphological characteristics. The culturing method of the present invention is different from other culturing methods of bacteria, and the main features are as follows.
(1) Divide the collected tissues or spores of wild strains.
(2) A single fungal colony or a single spore is purified and cultured.
(3) by observing the hyphae with a microscope, dividing the strain that matches the characteristics of the purified Kyushu Mushikusakin, making liquid bacterial species using the same.
(4) A lyophilized powder of pea larva larvae bred under aseptic conditions is mixed with cereals, and the above-mentioned liquid bacterial species are planted in a mixed culture medium to which nutrient water is added . Infect naturally until the mixed culture medium is filled with mycelium. Under appropriate conditions, it can grow into the same fruiting body as the wild strain.
[0015]
Here, the liquid species are 3 to 5 parts by weight of lyophilized powder of 3-5 year old larvae bred under aseptic conditions, 10 to 15 parts by weight of potato, 1-2 parts by weight of sugar, vitamin B 1 The purified strain is planted in a culture solution containing 0.001 to 0.002 parts by weight of the remainder, the remainder being water and the whole being 100 parts by weight.
[0016]
The mixed culture medium is 3 to 5 parts by weight of freeze-dried powder of 3 to 5 years old bean larvae bred under aseptic conditions, 97 to 95 parts by weight of grains such as rice and corn powder, and the total amount of these materials It is the composition which added 1.4 times the nutrient water. Here, nutrition water, 1.5 parts by weight of powder of purified Kyushu Mushikusakinko entity, 2-2.5 parts by weight of KH 2 PO 4, K 2 HPO 4 and 1.5 parts by weight , MgSO 4 is contained in an amount of 1 to 1.5 parts by weight, with the balance being water and the whole being 1000 parts by weight.
[0017]
According to the culture method of the present invention, Kyushu Mushikusa bacteria purified or pure Kyushu Mushikusakin in the process of selecting not only the to select, as made from artificial culture conditions Kyushu Mushikusa mycelium entity is the same as that of the wild Can be guaranteed. Compared with other artificial culture methods, the production cycle is short, the strain is stable, the inoculation efficiency is clearly increased, the survival rate and fruiting body production rate is 100% or more, and the transformation rate is 168.3. There are many advantages of being%, it can be the foundation can be continuously large scale production. Moreover, the tendency to inhibit the activity of the drug can also be prevented. The Shenyang Institute for Applied Ecology, CAS and Liaoning China New Drug Development Research Institute have reported that Kyushu insect fungi are similar in appearance to the wild, and the results are shown in Table 1.
[0018]
[Table 1]
In addition, Table 2 shows nutritional components of the fruit body of Kyushu insect fungus.
[0020]
[Table 2]
From these points, it turns out that the density | concentration of an active nutrient component has reached the high level. By producing health products using Kyushu Mushikusa mycelium entity as a material for drugs, it nourishes the lung, good kidney, hemostasis, improving degrade sputum, and health, weakened lungs and kidneys Can strengthen the ability to cure the disease caused by. In addition, normalization of the tumor (improved immunoregulatory function), expelling bad things (suppression of tumor growth and prolongation of life), reduction of toxicity (decrease of function of hematopoietic cells in medical treatment) It can also be used in such aspects, suppresses the growth of tumor cells, recovers from fatigue, and improves the body's immune function.
[0022]
Since the effective nutrient components of Kyushu insect fungus fruiting bodies are relatively high, even a small amount of material can reduce costs without impairing pharmacological functions. For this reason, the method described above has been treated as a major item studied in the science and technology sector and has been studied as the first class of new drugs.
[0023]
In order to put this to practical use, the Kyushu insect fungus fruiting body according to the present invention is dried, pulverized to a powder of 100 mesh, granulated with alcohol, dried at 60 ° C. or lower, and put into a capsule sterilized with Co60. Alternatively, mannitol, magnesium stearate, sodium saccharin, menthol and the like can be blended with Kyushu insect fungus body, granulated with alcohol, and dried to obtain tablets containing Kyushu insecticide. In addition to capsules and tablets, it can be used as an active ingredient in edible drugs, health foods, oral fluids, beverages and alcoholic beverages. With a small amount of material, its function is remarkable.
[0024]
(Example 1)
The outer capsule of the collected wild Kyushu insecticide fruit bodies was peeled off, and 0.2 to 0.3 cm of tissue was cut with a sterilized knife and placed on a culture dish containing a sterilized medium to inoculate the surface. After culturing at an appropriate temperature for 7 days, the strong mycelium was taken out and placed on the slope of the test tube, further cultured for 7 to 10 days, roughly selected and planted in a culture dish based on the growth state of the mycelium . Coverslips were inserted into all around the planting portion, the mycelium on the cover glass after 5 days were observed with a microscope, and the hyphae that fits the characteristics of the strains and liquid species. In addition, the liquid bacterial species is 10 mg of pure strain, freeze-dried powder of larvae of beans, 3-5 parts by weight , potato: 10-15 parts by weight , sugar: 1-2 parts by weight , vitamin B 1 : 0.001 to 0.002 parts by weight and the rest is water, and the whole is planted in a culture solution of 100 parts by weight.
[0025]
(Example 2 )
3 kg of lyophilized powder of Mametengu larvae, 10 kg of juzudama, 12 kg of ground corn, and 75 kg of rice were uniformly mixed, and 30 g of this mixture and 42 ml of nutrient water were packed in a plastic bottle. After sterilization, 2 ml of liquid fungus seeds were planted and cultured at an appropriate temperature for about 20 days . When the surface of the medium was filled with mycelium extending downward, it was ventilated by applying dispersed light (about 2500 Lux). (Membrane is attached to the edge of the bottle.) Doors and windows were opened 3-4 times a day for 20-30 minutes each time . Humidity in the room was 85% to 90%. After 25 to 30 days, fruiting bodies were obtained.
[0026]
(Example 3 )
4 kg of freeze-dried powder of pea tengu larvae, 9 kg of juzudama, 11 kg of crushed corn and 76 kg of rice were uniformly mixed, and 40 g of this mixture and 56 ml of nutrient water were packed in a bottle . After sterilization, 3 ml of liquid fungus seeds were planted and cultured for about 23 days at a suitable temperature without light. Once full to become in the mycelia surface of the medium is extended to lower, given the dispersed light.
[0027]
(Example 4 )
5 kg of beans larva powder, 8 kg of juzudama, 10 kg of crushed corn and 77 kg of rice were uniformly mixed, and 50 g of this mixture and 70 ml of nutrient water were packed in a bottle. After sterilization, 4 ml of liquid fungus seeds were planted and cultured at an appropriate temperature for about 25 days. When the surface of the medium was filled with the mycelium extending downward, scattered light was applied .
[0028]
【The invention's effect】
According to the present invention, from a strain obtained by purifying a single spore or colony of wild Kyushu insecticide, it is naturally infected in an arbitrary culture state until a fruiting body is finally formed. Incubation of fruiting bodies that are stable without causing degeneration, increase in infection efficiency, have a survival rate and a fruiting body generation rate of 100% or more, and a transformation rate of 168.3%. And can enable continuous large-scale production. Moreover, when this fruit body is used as a medicine material, it can be made into an effective drug in a small amount, and can be used as an active ingredient for health foods, beverages and the like .
[Brief description of the drawings]
FIG. 1 is a morphological view of a wild strain.
Figure 2 is a morphological view of cultured strains in culture medium made from Mameten moth larvae.
3 is a morphological view of cultured strains in culture medium made from Mameten moth larvae.
Claims (3)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN00110494A CN1109103C (en) | 2000-06-06 | 2000-06-06 | Artificial culture of jiuzhou caterpillar fungus and the application of its sporophore |
| CN00110494.2 | 2000-06-06 |
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| Publication Number | Publication Date |
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| JP2002045035A JP2002045035A (en) | 2002-02-12 |
| JP3665845B2 true JP3665845B2 (en) | 2005-06-29 |
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| JP2001171804A Expired - Fee Related JP3665845B2 (en) | 2000-06-06 | 2001-06-06 | Artificial culture method of Kyushu insecticide |
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| JP (1) | JP3665845B2 (en) |
| KR (1) | KR100442766B1 (en) |
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| CN100434508C (en) * | 2006-11-08 | 2008-11-19 | 中山大学 | Method for increasing yield of Qinghai Tibet plateau cordyceps |
| CN101659578B (en) * | 2009-09-18 | 2012-10-17 | 鲁东大学 | Artificial culture method of Kyushu cordyceps sinensis and culture medium thereof |
| CN104871830B (en) * | 2015-06-15 | 2017-03-15 | 鲁东大学 | With bean worm as the method for carrier-planting Cordyceps militaris (L.) Link. |
| CN110786204A (en) * | 2019-11-28 | 2020-02-14 | 张翰熙 | Industrial production method of cordyceps sinensis |
| CN112889758A (en) * | 2021-01-27 | 2021-06-04 | 连云港市农业科学院 | Environmental control condition for artificial indoor culture of bean hawkmoth |
| CN114568208B (en) * | 2022-03-14 | 2023-03-14 | 刘晓红 | Artificial cultivation method for cordyceps sinensis sporocarp |
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| JP2002045035A (en) | 2002-02-12 |
| CN1274751A (en) | 2000-11-29 |
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