JPS611379A - Novel fungal strain resulting from cellfusion between edible fungi: "matsutake" and "shiitake" - Google Patents

Abstract

PURPOSE:Cell fusion between MATSUTAKE (a kind of mushroom: Armillaria matsudake) and SHIITAKE (a kind of mushroom: Cortinellus shiitake) is effected to form hybrid cells and they are cultured to obtain a new type of fungus which has good smell and shape as well as properties of wood-decaying fungi. CONSTITUTION:Tissue pieces of fruit bodies of MATSUTAKE and SHIITAKE are subjected to sterile culture, respectively to make mycelia grow. The mycelia are multiplied and transplanted in a liquid medium to effect subcultivation. Then, the culture mixtures are treated with enzyme solutions, respectively to separate protoplasts of individual fungi. The enzyme solutions containing the protoplast is centrifuged and washed, both suspensions of SHIITAKE and MATSUTAKE are mixed and centrifuged and the residue is dispersed again in a polyethylene glycol. Further, a culture medium for cell fusion is added to the suspension and they are cultured to give the objective new type of fungus.

Description

【発明の詳細な説明】 〔産業上の利用分野〕 本発明はマツタケとシイタケの細胞融合を用いて得らｎ
る新菌種に関するものである。[Detailed Description of the Invention] [Industrial Field of Application] The present invention is directed to the use of matsutake mushroom and shiitake mushroom cell fusion.
This study concerns new bacterial species.

〔従来の技術及び発明が解決しようとする問題点〕マツ
タケは古来、その香り、味が珍重さｎてき友が、菌根菌
であるため、その栽培は不可能とさｎて来たのに対し、
シイタケは木材腐釘菌の性質が利用され、その商業的栽
培が行なわｎ現在では周知の日常的食料となっている。[Problems to be solved by conventional techniques and inventions] Since ancient times, matsutake mushrooms have been prized for their aroma and taste, but because they are mycorrhizal fungi, it has been considered impossible to cultivate them. On the other hand,
Shiitake mushrooms have been cultivated commercially by utilizing the properties of wood-rotting fungi, and are now a well-known daily food.

本発明者らは、との両画類の性質を相補的に併わせもつ
新菌種を得るために、細胞融合の手法に着目し、鋭意研
究を重ねた結果、前記の性質を併有する新菌種の育種に
成功し本発明を完成するに至った。The present inventors focused on cell fusion techniques and conducted extensive research in order to obtain a new bacterial species that has the properties of both classes in a complementary manner. They succeeded in breeding the fungal species and completed the present invention.

〔問題点を解決するための手段〕[Means for solving problems]

すなわち本発明はマツタケとシイタケの細胞融合により
雑種細胞を得、こｎを培養することにより得ら几る新菌
種であって、マツタケの香り、形態と、木材腐打菌の性
質を併有する新菌種を提供するものである。以下、本発
明を更に具体的に説明する。That is, the present invention is a new fungal species obtained by obtaining hybrid cells by cell fusion of matsutake and shiitake and culturing them, which has both the aroma and morphology of matsutake and the properties of wood rot fungi. It provides new bacterial species. The present invention will be explained in more detail below.

（笑施例） まず、マツタケ、シイタケの子実体の組織片を各々表１
の組成よりなる培地に無菌的に植え込み、菌糸を生育さ
せる。(Example) First, tissue pieces of the fruiting bodies of Matsutake and Shiitake were prepared in Table 1.
The mycelium is grown aseptically in a medium with the following composition.

この菌糸を増殖させた後、各々先述の組成の液体培地に
移しかえ、継代培養の後、植え継ぎ後、約２週間の菌糸
を各々、表２．３の組成の酵素液で２５℃３時間処理す
ることにより各々両画類のプロトプラストが分離される
。このプロトプラストを含む酵素液を　７００ＸＰで遠
心分離し、上澄を捨てる。After growing these hyphae, they were each transferred to a liquid medium with the composition described above, and after subculturing, the hyphae, which had been about 2 weeks old after subculturing, were treated with an enzyme solution having the composition shown in Table 2.3 at 25°C. By time processing, protoplasts of both classes are separated. Centrifuge the enzyme solution containing protoplasts at 700XP and discard the supernatant.

残渣に０．７　Ｍ　Ｎａｃｊ’　水溶′ｆｅＬを加えて
再懸濁し、上記条件にて遠心分離を行ない上澄を捨てる
。この洗浄操作を３回くり返した後、各々１．０Ｍマン
ニトール水溶液にグロトプラスト濃度が１〜５ＸｌＯ’
個／ｄＫなるように再懸濁する。Add 0.7 M Nacj'feL in water to resuspend the residue, centrifuge under the above conditions, and discard the supernatant. After repeating this washing operation three times, the grotoplast concentration was 1 to 5XlO' in each 1.0M mannitol aqueous solution.
Resuspend to a concentration of 100 μg/dK.

このようにして調製したマツタケ、シイタケの両懸濁液
を混合し、７０（ＩＸＦ　で遠心分離し、上澄を捨て、
残漬を３０℃に保った表４の組成のポリエチレングリコ
ール溶液に再懸濁し１０分間放置する。このＳ濁液に表
５の組成の融合細胞培養培地を十分量加え、７００　Ｘ
Ｆで５分間遠心し、上置を捨てる。Both matsutake and shiitake suspensions prepared in this way were mixed, centrifuged at 70°C (IXF), and the supernatant was discarded.
The residue was resuspended in a polyethylene glycol solution having the composition shown in Table 4 kept at 30°C and left for 10 minutes. Add a sufficient amount of fused cell culture medium with the composition shown in Table 5 to this S suspension, and incubate at 700X.
Centrifuge at F for 5 minutes and discard the supernatant.

残漬を同様の操作により２度洗浄し几後、融合細胞培養
培地に再懸濁し、シャーレ上に移し、得られた融合細胞
をマイクロマニュプレーターにてピックアラブレ、融合
細胞用培地で２５℃暗黒下にて培養する。培養約１ケ月
後にこの雑種細胞は***をおこない、菌糸の生長とコロ
ニー形成が観原さｎｔ−Ｑ 表１ （効果） 上記のようにして得らｎた本発明の新菌種は増殖、徨ゴ
マ作裂、植菌、子実体形成等の過程を経て、マツタケの
香りと形態を有するキノコが生産される。しかも本発明
の新菌種はシイタケと同様木材屑Ａ５菌の性質を有する
ので、商業的栽培が可能で６９、その効果は極めて大き
い。The remaining cells were washed twice in the same manner, and after cooling, they were resuspended in a fused cell culture medium and transferred onto a Petri dish. The obtained fused cells were picked and washed with a micromanipulator and incubated in the dark at 25°C in a fused cell culture medium. Culture at the bottom. After about one month of culture, the hybrid cells divide, and hyphal growth and colony formation are observed. Through processes such as sesame seed splitting, inoculation, and fruiting body formation, mushrooms with the aroma and shape of matsutake are produced. Moreover, since the new fungal species of the present invention has the properties of wood waste A5 fungi like shiitake mushrooms, it can be cultivated commercially69, and its effects are extremely large.

【図面の簡単な説明】[Brief explanation of the drawing]

第１図はマツタケのグロトプラストの顕微鏡写真、第２
図はシイタケのグロトプラストのｋＥ８４倣境写真、第
３図はマツタケシイタケ両グロトプラストの融合細胞の
顕微鏡写真である０図面１．″）浄書（内容に変更、ｚ
・シ）ミ ； し、 ；ミ へ ゛≧ 手続補正書く方式） １．事件の表示 昭和５９年特許願　第１２０８４８号 ２、発明の名称 マツタケとシイタケの細胞融合による新菌種３、補正を
する者 （事件との関係）　特許出願人 住　所　　大阪府大阪市大淀区豊崎３丁目２１番３号４
、補正命令の日付（発送困 昭和５９年９月２５日 ５、補正の対象 第１図は、マツタケのプロトプラストを示す顕微鏡写真
（１７５０倍）でらる０ 第２図は、シイタケのプロトプラストを示す顕微鏡写真
（１７５０倍）であるっ 第３図は、マツタケ　シイタケ両プロトプラストの融合
細胞を示す顕微鏡写真（３５嘩）である、尚、第１〜第
３図は、生物の形態に関する図面に代る顕微鏡写真であ
る。Figure 1 is a microscopic photograph of the glotoplast of Matsutake, Figure 2
The figure is a kE84 imitation photograph of a grotoplast of a shiitake mushroom, and Figure 3 is a micrograph of a fused cell of both glotoplasts of a shiitake mushroom. ″) engraving (change to content, z
・Sh)Mi; Shi, ;Mihe゛≧ Procedure amendment writing method) 1. Indication of the case: 1982 Patent Application No. 120848 2, Name of the invention: New bacterial species resulting from cell fusion of matsutake and shiitake mushrooms 3, Person making the amendment (relationship with the case) Patent applicant address: Toyosaki, Oyodo-ku, Osaka-shi, Osaka Prefecture 3-21-3-4
, date of amendment order (shipped September 25, 1982), subject of amendment Figure 1 is a micrograph (1750x) showing protoplasts of matsutake mushrooms Figure 2 shows protoplasts of shiitake mushrooms Figure 3 is a micrograph (1750x) showing a fused cell of Matsutake and Shiitake protoplasts. Figures 1 to 3 are diagrams showing the morphology of the organism. This is a microscopic photograph.

Claims (1)

【特許請求の範囲】[Claims] マツタケとシイタケの細胞融合により雑菌細胞を得、こ
れを培養することにより得られた新菌種A new bacterial species obtained by culturing various bacterial cells obtained by cell fusion of matsutake and shiitake