JP2007053928A - New fungal strain and method for culturing new fruit body - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new fungal strain belonging to the genus Pleurotus without requiring a complex temperature change-controlling process as adopted in the culture of Pleurotus nebrodensis and capable of easily culturing by adopting a conventional Pleurotus eryngii culturing technology as it is. <P>SOLUTION: This method for obtaining a KX-EN2001 fungal strain (FERM P-20576) which belongs to the fungal strain of the genus Pleurotus is provided by crossing the spores of the Pleurotus eryngii with those of the Pleurotus nebrodensis, and then culturing the crossed fungi by using a Pleurotus eryngii-culturing medium obtained by mixing saw dust powder and nutrition sources, and adjusting water content. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

本発明は、新規菌株の作出方法および新規菌株ならびに新規子実体の栽培方法に関し、詳しくは、食用きのことして有用なプレオロウタス属に属する新規菌株の作出方法および新規菌株：ＫＸ−ＥＮ２００１菌株（ＦＥＲＭ Ｐ−２０５７６）並びに新規子実体の栽培方法に関する。   The present invention relates to a method for producing a new strain and a method for cultivating a new strain and a new fruiting body, and more particularly, a method for producing a new strain belonging to the genus Pleurotus useful as an edible mushroom and a novel strain: KX-EN2001 strain (FERM P- 20576) and a method for cultivating new fruiting bodies.

近年、中国などから導入された「白霊茸（Pleurotus nebrodensis）」が日本においても人工栽培されるようになってきている。具体的には、適宜に調整した培養基に白霊茸（Pleurotus nebrodensis）の種菌を接種し、培養前期の温度を２０℃前後に４０日間維持した後、培養後期において培養前期に維持していた温度を１０日間程度１０℃前後に一旦低下させ、その後の１０日間を３０℃前後まで著しく上昇させるようにして培養基内に菌糸を蔓延させた後、発生時においては、前期の数日間を−１℃前後と低温に維持し、その後５℃前後で７日間程度維持した後に菌掻きを行い、菌掻き後に温度を１８℃前後に上昇させて発生させる栽培方法が提案されている（特許文献１）。
特開２００４−２０１６４５号 In recent years, “Pleurotus nebrodensis” introduced from China and other countries has been artificially cultivated in Japan. Specifically, the inoculum of Pleurotus nebrodensis was inoculated into an appropriately adjusted culture medium, and the temperature maintained in the first culture period in the second culture period after maintaining the temperature in the first culture period at around 20 ° C. for 40 days. For about 10 days, the hyphae are spread in the culture medium so that the subsequent 10 days are remarkably raised to about 30 ° C., and at the time of development, the first few days are reduced to −1 ° C. A cultivation method has been proposed in which the temperature is maintained at around and at a low temperature and then maintained at around 5 ° C. for about 7 days, and then the bacteria are scraped, and the temperature is raised to around 18 ° C. after the bacteria are scraped (Patent Document 1).
JP 2004-201645 A

しかしながら、上記のような白霊茸（Pleurotus nebrodensis）の栽培においては、従来のエリンギ（Pleurotus eryngii）の栽培に比べ栽培サイクルが長く、しかも栽培工程において−１〜１０℃までの変温管理による低温ショックを付与しなければきのこが発生しない等の栽培工程上の問題点が多く、既存施設での新規の栽培きのことしての導入においては経営上のメリットのないことが欠点となっている。   However, in the cultivation of white leopard (Pleurotus nebrodensis) as described above, the cultivation cycle is longer than in the conventional cultivation of eringi (Pleurotus eryngii), and in the cultivation process, the temperature is low due to temperature-change management up to -1 to 10 ° C. There are many problems in the cultivation process such that mushrooms do not occur unless a shock is applied, and there is a disadvantage that there is no management merit in introducing new cultivation mushrooms in existing facilities.

本発明は上記の実情に鑑みなされたものであり、その目的は、白霊茸（Pleurotus nebrodensis）で採用されるような複雑な変温管理工程を必要とせず、従来のエリンギ栽培技術をそのまま採用することで容易に栽培が可能なプレオロウタス属に属する新規な菌株を開発することにある。   The present invention has been made in view of the above circumstances, and the purpose thereof is not to require a complicated temperature change management process such as that employed in white leopard (Pleurotus nebrodensis), and the conventional eringi cultivation technique is adopted as it is. This is to develop a new strain belonging to the genus Pleurotus that can be easily cultivated.

本発明者らは、種の異なるプレオロウタス属のきのこに属するエリンギ「Pleurotus eryngii（DC.:Fr.）Quel.」とその変種である白霊茸「Pleurotus nebrodensis（Inzenga）Quel.」とが交配する事実を見出し、その交配させた新菌株を使用してエリンギ栽培用の人工培養基を使用して交配により作出した菌株の選抜を行った結果、エリンギと同様の栽培方法で栽培が可能なプレオロウタス属に属する新規な菌株と栽培法を見出し、本発明を完成した。   The present inventors mated an eringi “Pleurotus eryngii (DC .: Fr.) Quel.” Belonging to mushrooms of different species of Pleurotus and a variant white phoenix “Pleurotus nebrodensis (Inzenga) Quel.” As a result of finding the facts and selecting the strains produced by mating using the artificial culture medium for growing eringi using the new bred strain, it was found that the genus Pleurotus can be cultivated by the same cultivation method as eringi. The present inventors have found a novel strain and cultivation method to which the present invention belongs, and have completed the present invention.

すなわち、本発明の第１の要旨は、エリンギの胞子と白霊茸の胞子とを交配させた後、オガコと栄養源とを混合して水分調整したエリンギ栽培用培養基で交配菌を培養して選抜を行うことを特徴とする新規菌株の作出方法に存する。   That is, the first gist of the present invention is to cultivate a mating bacterium in a culture medium for cultivating eringi mixed with sawdust and nutrients after mating spore of eringi and spore of white mackerel. It exists in the production method of the novel strain characterized by performing selection.

本発明の第２の要旨は、プレオロウタス属菌株であるＫＸ−ＥＮ２００１菌株（ＦＥＲＭ Ｐ−２０５７６）に存する。   The second gist of the present invention resides in the strain KX-EN2001 (FERM P-20576), which is a strain of the genus Pleurotus.

そして、本発明の第３の要旨は、栽培容器にオガコと栄養源とを混合して水分調整した培養基を充填し、培養基を加熱殺菌した後、請求項１で得られた新規菌株の種菌を接種し、芽出室に搬入し、芽出しを行ない、次いで、原基の生育を行なうことを特徴とする新規子実体の栽培方法に存する。   The third gist of the present invention is that the cultivation container is filled with the culture medium prepared by mixing the sawdust and the nutrient source, and the culture medium is heated and sterilized, and then the inoculum of the novel strain obtained in claim 1 is obtained. A new fruiting body cultivation method characterized by inoculating, carrying into a sprouting chamber, sprouting, and then growing a primordial base.

本発明によれば、エリンギ（Pleurotus eryngii）と白霊茸（Pleurotus nebrodensis）とを交配させて作出した新規な菌株を使用することにより、エリンギ（Pleurotus eryngii）と同様の人工栽培方法で新規な菌株の栽培を行うことが出来るため、白霊茸（Pleurotus nebrodensis）のような多段階の変温管理工程を必要とすることなく、プレオロウタス属に属する新規なきのこを容易に栽培することが出来る。   According to the present invention, by using a novel strain produced by crossing eringi (Pleurotus eryngii) and white leopard (Pleurotus nebrodensis), a novel strain can be obtained by an artificial cultivation method similar to that of eringi (Pleurotus eryngii). Therefore, a new mushroom belonging to the genus Pleurotus can be easily cultivated without requiring a multi-stage temperature change management process such as white leopard (Pleurotus nebrodensis).

＜新規菌株の作出方法＞
本発明に係る新規菌株の作出方法は、エリンギの胞子と白霊茸の胞子とを交配させた後、オガコと栄養源とを混合して水分調整したエリンギ栽培用培養基で交配菌を培養して選抜を行うことを特徴とする。 <New strain production method>
In the method for producing a novel strain according to the present invention, after mating spore of eringi and spore of white mausoleum, the mating bacterium is cultured in a culture medium for cultivating eringi mixed with sawdust and nutrient sources to adjust the water content. It is characterized by performing selection.

交配に使用するエリンギ「Pleurotus eryngii（DC.:Fr.）Quel.」と白霊茸「Pleurotus nebrodensis（Inzenga）Quel.」は、その組み合わせであれば、特に親株が限定されるものではない。通常、交配は次に様に行われる。   The parent strain is not particularly limited as long as it is a combination of the eringi “Pleurotus eryngii (DC .: Fr.) Quel.” And the white mausoleum “Pleurotus nebrodensis (Inzenga) Quel.” Used for mating. Usually, mating is performed as follows.

先ず、落下法により、エリンギ子実体と白霊茸の傘をペトリー皿に入れて静置し、得られた胞子紋に滅菌水を加えて胞子を懸濁させる。血球計算板を使用し、胞子懸濁液中の胞子数を計数後、適当な胞子濃度に希釈し、ＰＤＡ（ポテト・デキストロース寒天）平板培地に接種する。常温で７〜１０日間培養した後、発芽した１次菌糸を分離し、各々の１次菌糸株を得る。次いで、ＰＤＡ平板培地に両１次菌糸株を接種し、常温で７〜３０日間培養し、総当りの交配を行う。交配の有無は、光学顕微鏡下でクランプ結合を確認することにより行う。   First, by the dropping method, an eringi fruit body and a white mausoleum umbrella are placed in a petri dish and allowed to stand, and spore is suspended by adding sterile water to the obtained spore pattern. Using a hemocytometer, count the number of spores in the spore suspension, dilute to an appropriate spore concentration, and inoculate on a PDA (potato dextrose agar) plate medium. After culturing at room temperature for 7 to 10 days, the germinated primary mycelium is separated to obtain each primary mycelium strain. Subsequently, both primary mycelia strains are inoculated on a PDA plate medium, cultured at room temperature for 7 to 30 days, and cross-bred. The presence / absence of mating is confirmed by confirming clamp bonding under an optical microscope.

交配菌の選抜は、適当な培養基を使用して行う。培養基としては、エリンギ栽培の可能な培養基であれば何れの培養基であってもよい。一般的には木粉培養基が使用され、好ましくはエリンギ栽培用培養基、すなわち、オガコと栄養源とを混合して水分調整したエリンギ栽培用培養基が使用される。培養基の調整に使用されるオガコとしては、針葉樹オガコが好適であるが、広葉樹オガコも使用することが出来る。また、３ヶ月以上堆積した針葉樹オガコが好適であるが、新鮮な針葉樹オガコも使用することが出来る。オガコと共にコーンコブ粉砕物やコットンハル等の代替培地基材を使用することも出来る。その場合、オガコ：コーンコブ粉砕物の容積比は、通常１０：０〜０：１０、好ましくは８：２程度とされる。一方、栄養源としては、米糠、フスマ、大麦糠、トウモロコシ糠などの穀類糠が好適に使用される。穀類糠は、出来る限り新鮮なものが好ましい。培養基総重量に対し、栄養源の使用割合は、通常０〜３０重量％、好ましくは１２〜１６重量％（８５０cc１ビン当たり７０〜１００ｇ）の範囲とされる。木粉培養基の含水率は、通常５５〜７５重量％、好ましくは６４〜６８重量％の範囲とされる。   Selection of mating bacteria is performed using an appropriate culture medium. As a culture medium, any culture medium may be used as long as it is a culture medium capable of cultivating eringi. In general, a wood flour culture medium is used, and preferably a culture medium for cultivating eringi, that is, a culture medium for cultivating eringi in which moisture is adjusted by mixing sawdust and nutrient sources. The sawdust used for adjusting the culture medium is preferably a coniferous saw, but a broadleaf saw can also be used. Moreover, although the coniferous sawdust accumulated for three months or more is suitable, a fresh coniferous sawfly can also be used. Alternative medium base materials such as corn cob pulverized material and cotton hull can be used together with sawdust. In this case, the volume ratio of the sculpture of corncob: corn cob is usually 10: 0 to 0:10, preferably about 8: 2. On the other hand, cereal meal such as rice bran, bran, barley meal and corn meal is preferably used as a nutrient source. The cereal meal is preferably as fresh as possible. The ratio of the nutrient source to be used is usually 0 to 30% by weight, preferably 12 to 16% by weight (70 to 100 g per 850 cc bottle) with respect to the total weight of the culture medium. The moisture content of the wood flour culture medium is usually 55 to 75% by weight, preferably 64 to 68% by weight.

上記の木粉培養基による培養（選抜）は、二段階以上に分け、木粉培地適応性を有する交配菌を選抜し、木粉培地に順応性を有する交配菌について更に選抜を行うことが出来る。この場合、第１段目の選抜と第２段目の選抜とで使用する培養基の組成は異ならせることが出来、第１段目の選抜の培養基には栄養源の配合を省略することも出来る。また、第２段目の選抜の選抜は、第１段目の選抜の培養基で製造した菌床を種菌として使用して行われる。そして、最終選抜は、培養の終了した菌床の表面を約１５ｍｍの深さに掻き取り、倒立状態で環境温度１５〜１６℃、炭酸ガス濃度８００〜２，０００ｐｐｍ、昼間の時間帯のみ４００Ｌｘ程度の光を照射し、環境湿度７０〜９８％の湿度範囲で低湿度環境と高湿度環境とを一定間隔で繰り返しながら芽出し管理を行い、芽切り確認後に容器を正立状態に戻し、環境温度を１６〜１８℃、環境湿度を８０〜９０％に変更し、芽出し管理と同様の照度環境下で生育管理を行うという、一般的なエリンギの栽培管理で行う。   The culture (selection) using the above-mentioned wood flour culture medium can be divided into two or more stages, selecting a mating bacterium having adaptability to the wood powder medium, and further selecting the mating bacteria having adaptability to the wood powder medium. In this case, the composition of the culture medium used in the selection of the first stage and the selection of the second stage can be different, and the composition of the nutrient source can be omitted in the culture medium of the first stage selection. . In addition, selection in the second stage selection is performed by using a fungal bed produced with the culture medium in the first stage selection as an inoculum. And the final selection is to scrape the surface of the cultivated bacterial bed to a depth of about 15 mm, and in an inverted state, the ambient temperature is 15 to 16 ° C., the carbon dioxide concentration is 800 to 2,000 ppm, and the daytime is only about 400 Lx. The sprouting management is performed while repeating the low humidity environment and the high humidity environment at regular intervals in the humidity range of 70 to 98%, and after confirming the sprouting, the container is returned to the upright state, and the environmental temperature is adjusted. It is carried out in general cultivation management of eringi, in which the growth is managed under an illuminance environment similar to the germination management by changing the ambient humidity to 16 to 18 ° C. and 80 to 90%.

＜新規菌株＞
本発明者らは、従来のエリンギ栽培技術をそのまま採用することで容易に栽培が可能な新規な菌株を見出し、ＫＸ−ＥＮ２００１と命名した。このＫＸ−ＥＮ２００１株は、プレオロウタス属に属する新規な菌株であり、平成１７年６月２９日から、独立行政法人産業技術総合研究所に１７産生寄第８８号（ＦＥＲＭ Ｐ−２０５７６）として寄託されている。本発明の菌株の菌学的諸性質は次の（１）〜（１１）に示す通りである。 <New strain>
The present inventors have found a novel strain that can be easily cultivated by adopting conventional eringi cultivation technology as it is, and named it KX-EN2001. This KX-EN2001 strain is a novel strain belonging to the genus Pleurotus, and has been deposited as an Independent Administrative Institution National Institute of Advanced Industrial Science and Technology as No. 88 Production No. 88 (FERM P-20576) from June 29, 2005. ing. The mycological properties of the strain of the present invention are as shown in the following (1) to (11).

（１）麦芽エキス寒天培地における生育状態（２３℃）：
７日目のコロニー直径は２６．８ｍｍ。菌糸は白色で密。気中菌糸の量は中程度で、直線的に伸びる。 (1) Growth state in malt extract agar medium (23 ° C.):
The day 7 colony diameter was 26.8 mm. Mycelium is white and dense. The amount of aerial hyphae is moderate and grows linearly.

（２）バレイショ・・ブドウ糖寒天培地における生育状態（２３℃）：
７日目のコロニー直径は３０．４ｍｍ。菌糸は白色で密。気中菌糸の量は中程度で、直線的に伸びる。 (2) Growth state in potato / glucose agar medium (23 ° C.):
The colony diameter on the 7th day is 30.4 mm. Mycelium is white and dense. The amount of aerial hyphae is moderate and grows linearly.

（３）ツアペック・・ドックス寒天培地における生育状態（２３℃）：
７日目のコロニー直径は１３．２ｍｍ。菌糸は白色で極めて希薄。気中菌糸の量は極めて少なく、直線的に伸びる。 (3) Growth state on the Tuapekk Docs agar medium (23 ° C.):
The day 7 colony diameter was 13.2 mm. Mycelium is white and extremely dilute. The amount of aerial hyphae is very small and grows linearly.

（４）サブロー寒天培地における生育状態（２３℃）：
７日目のコロニー直径は２９．９ｍｍ。菌糸は白色で密。気中菌糸の量は中程度で、直線的に伸びる。 (4) Growth state on Sabouraud agar medium (23 ° C.):
The day 7 colony diameter was 29.9 mm. Mycelium is white and dense. The amount of aerial hyphae is moderate and grows linearly.

（５）オートミール寒天培地における生育状態（２３℃）：
７日目のコロニー直径は２８．５ｍｍ。菌糸は白色で希薄。気中菌糸の量は極めて少なく、直線的に伸びる。 (5) Growth state in oatmeal agar medium (23 ° C.):
The day 7 colony diameter was 28.5 mm. Mycelium is white and thin. The amount of aerial hyphae is very small and extends linearly.

（６）合成ムコール寒天培地における生育状態（２３℃）：
７日目のコロニー直径は５．５ｍｍ。菌糸は白色で密。気中菌糸の量が多く、樹状に伸びる。 (6) Growth state on synthetic mucor agar medium (23 ° C.):
On the 7th day, the colony diameter was 5.5 mm. Mycelium is white and dense. The amount of aerial hyphae is large and grows in a tree shape.

（７）ＹｐＳｓ寒天培地における生育状態（２３℃）：
７日目のコロニー直径は３８．８ｍｍ。菌糸は白色で密。気中菌糸の量は中程度で、直線的に伸びる。 (7) Growth state on YpSs agar medium (23 ° C.):
The day 7 colony diameter was 38.8 mm. Mycelium is white and dense. The amount of aerial hyphae is moderate and grows linearly.

（８）フェノールオキシダーゼ検定培地における生育状態（２３℃）
０．５％タンニン酸と０．５％没食子酸添加ＰＤＡ培地、および０．５％没食子酸添加麦芽エキス寒天培地のいずれの培地とも、菌糸は活着、生長せずに不活着症状を示し、接種源の周辺培地が少し褐変する症状が認められる。 (8) Growth state in phenol oxidase assay medium (23 ° C.)
The mycelium is inactivated and does not grow and inoculates with both 0.5% tannic acid and PDA medium containing 0.5% gallic acid and 0.5% gallic acid added malt extract agar medium. Symptoms of browning of the medium surrounding the source are observed.

（９）菌糸生育最適温度：
ＰＤＡ培地上に直径５ｍｍの種菌を接種し、各温度帯でそれぞれ培養して、７日後に各コロニー径を測定したところ、菌糸伸長の最適温度は３０℃付近であった。 (9) Mycelium growth optimum temperature:
An inoculum with a diameter of 5 mm was inoculated on the PDA medium, cultured in each temperature zone, and each colony diameter was measured after 7 days. The optimum temperature for hyphal elongation was around 30 ° C.

（１０）菌糸生育最適ｐＨ：
ＹＧＴ液体培地（酵母エキス・ブドウ糖・チアミン塩酸培地）１６ｍｌずつを滅菌後、各ｐＨに調整し、種菌を接種後、２３℃で７日間静置培養した後、乾燥菌体重量を測定したところ、菌糸生育最適ｐＨは５．５付近であった。 (10) Optimum hyphal growth pH:
After sterilizing each 16 ml of YGT liquid medium (yeast extract / glucose / thiamine hydrochloride medium), adjusting to each pH, inoculating the inoculum, and then standing at 23 ° C. for 7 days, the dry cell weight was measured. The optimum pH for hyphal growth was around 5.5.

（１１）遺伝的特性：
遺伝的特性については、両菌糸が持っている性因子が異なれば、その菌糸は互いに異なる菌糸であるという菌類分類学的事実に基づき、両親株ならびに近縁品種との寒天培地上における対峙培養により性因子の同異判定を行った。 (11) Genetic characteristics:
As for genetic characteristics, based on the fungal taxonomic fact that, if the sex factors of both mycelia are different, the mycelia are different from each other, the parental strain and related varieties are cultured on the agar medium. Sex factor discrimination was performed.

供試したエリンギ「Pleurotus eryngii（DC.:Fr.）Quel.」の菌株としては、親株（ＫＸ−ＥＧ０７９号）の他にＡＴＣＣ３６０４７、ＡＴＣＣ９０２１２、ＡＴＣＣ９０７８７、ＡＴＣＣ９０８８７、ＡＴＣＣ９６０５４の各菌株を使用した。   As the strain of the tested eringi "Pleurotus eryngii (DC .: Fr.) Quel.", Each strain of ATCC36047, ATCC90212, ATCC90787, ATCC90887, and ATCC96054 was used in addition to the parent strain (KX-EG079).

白霊茸「Pleurotus nebrodensis（Inzenga）Quel.」の菌株としては、親株（中国遼寧省瀋陽農業大学所有の白霊茸）の他に新宇食用菌研究所所有白霊茸（Ｐｎｋｗ−１株）を使用した。   As a strain of the white mausoleum “Pleurotus nebrodensis (Inzenga) Quel.”, In addition to the parent strain (white mausoleum owned by Shenyang University of Agriculture, Liaoning Province, China) used.

また、プレオロウタス属の近縁株としては、Pleurotus ostreatus（DC.:Fr.）Quel.（ヒラタケ：弊社保有株）、Pleurotus pulmonarius（DC.:Fr.）Quel.（ウスヒラタケ：弊社保有株）、Pleurotus salmoneostramineus（DC.:Fr.）Quel.（トキイロヒラタケ：弊社保有株）、Pleurotus cornucopiae var. citrinopileatus（DC.:Fr.）Quel.（タモギタケ：弊社保有株）、Pleurotus djamor（Fries）Boedjin（ＮＢＲＣ３２３９８）、Pleurotus dryinus（Persoon:Fries）Kummer（ＮＢＲＣ３２７９７）、Pleurotus ferulae Lanzi（新宇食用菌研究所所有:Ｐｎｋｗ−４）を使用した。   Pleurotus ostreatus (DC.:Fr.) Quel. (Oyster mushrooms: our company), Pleurotus pulmonarius (DC .: Fr.) Quel. (Ushiratake: our company), Pleurotus salmoneostramineus (DC.:Fr.) Quel. ), Pleurotus dryinus (Persoon: Fries) Kummer (NBRC32797), Pleurotus ferulae Lanzi (owned by Shin Uedo Institute for Bacteria): Pnkw-4).

常法によりＰＤＡ寒天培地上に３ｃｍの間隔で上記の各菌株を接種し、２３℃で１４日間培養した後、両コロニー境界部に帯線が生じるか否かを判定した。結果を表１に示す（帯線形成は＋、形成しないものは−で表示）。   Each of the above strains was inoculated on a PDA agar medium at an interval of 3 cm by a conventional method, and cultured at 23 ° C. for 14 days. Then, it was determined whether or not a band was formed at the boundary between both colonies. The results are shown in Table 1 (indicated by + for forming a band, and-for those not formed).

表１に示す通り、前記の各菌株は、ＫＸ−ＥＮ２００１と明確な帯線を形成することから、当菌株はプレオロウタス属に属する新規な菌株であることが明らかである。   As shown in Table 1, each of the above-mentioned strains forms a clear band with KX-EN2001, and thus it is clear that this strain is a novel strain belonging to the genus Pleurotus.

＜新規子実体の栽培方法＞
本発明に係る新規子実体の栽培方法は、栽培容器にオガコと栄養源とを混合して水分調整した培養基を充填し、培養基を加熱殺菌した後、請求項１で得られた新規菌株の種菌を接種し、芽出室に搬入し、芽出しを行ない、次いで、原基の生育を行なうことを特徴とする。 <Cultivation method of new fruit body>
The cultivation method of the novel fruiting body which concerns on this invention fills the culture medium which mixed the sawfish and the nutrient source into the cultivation container, adjusted the water | moisture content, and heat-sterilized the culture medium, Then, the inoculum of the novel strain obtained in Claim 1 , Inoculated into the sprouting chamber, sprouting, and then growing the primordium.

上記の培養基としては、前述のエリンギ栽培用培養基を使用することが出来、栽培管理は、ポリプロピレン製栽培瓶（８５０cc）に正味重量で５３０〜５４０ｇの培養基を充填し、培養基の中央部に直径が約１５ｍｍで底部に到達する接種孔を設けて施栓し、常法に従って高圧殺菌釜中で殺菌処理を行った後、クリーンルーム内で無菌的に冷却して、プレオロウタス属に属する新規な菌株（ＫＸ‐ＥＮ２００１）を接種し、２３℃で３５日間の培養管理を行った。次いで、培養の終了した菌床の表面を約１５ｍｍの深さに掻き取り、倒立状態で環境温度１５〜１６℃、炭酸ガス濃度８００〜２，０００ｐｐｍ、昼間の時間帯のみ４００Ｌｘ程度の光を照射し、環境湿度７０〜９８％の湿度範囲で低湿度環境と高湿度環境とを一定間隔で繰り返しながら芽出し管理を行い、芽切り確認後に容器を正立状態に戻して、環境温度を１６〜１８℃、環境湿度を８０〜９０％に変更して、芽出し管理と同様の照度環境下で生育管理を行った。   As the above culture medium, the above-mentioned culture medium for cultivating eringi can be used, and the cultivation management is carried out by filling a polypropylene cultivation bottle (850 cc) with a culture medium of 530 to 540 g in net weight and having a diameter at the center of the culture medium. The inoculation hole reaching the bottom at about 15 mm is provided and plugged. After sterilization in a high-pressure sterilization pot according to a conventional method, it is cooled aseptically in a clean room, and a new strain belonging to the genus Pleurotus (KX- EN2001) was inoculated and the culture was controlled at 23 ° C. for 35 days. Next, the surface of the cultivated bacterial bed is scraped to a depth of about 15 mm, and in an inverted state, the ambient temperature is 15 to 16 ° C., the carbon dioxide concentration is 800 to 2,000 ppm, and the light of about 400 Lx is irradiated only during the daytime. Then, sprouting management is performed while repeating a low humidity environment and a high humidity environment at regular intervals in a humidity range of 70 to 98%, and after confirming sprouting, the container is returned to an upright state, and the environmental temperature is set to 16 to 18 The growth management was performed under the same illuminance environment as the germination management by changing the environmental humidity to 80 to 90%.

本発明の新規な菌株における子実体の形態的特性は次の通りである。すなわち、子実体は散生、菌傘の大きさは８〜１０ｃｍ、円形または不正形で丸ないし漏斗形、傘色は淡黄白色ないし灰黄色、表面は平滑で艶があり、筋状の紋様を有する。肉は白色かつ厚く緻密である。ヒダは淡灰黄色、密度は密生で、菌柄への付き方は強い垂生である。菌柄は中心ないし偏心生、太さは３〜４ｃｍ、長さ６〜８ｃｍで、色は白色、形は下太、肉質は白色、中実で非常に硬い。胞子は楕円ないし紡錘形、表面は平滑、大きさは５〜６×７〜９μｍで、胞子紋は白色である。   Morphological characteristics of fruit bodies in the novel strain of the present invention are as follows. That is, the fruiting body is scattered, the size of the fungus is 8-10 cm, round or irregular shape, round or funnel shape, the umbrella color is light yellowish white or grayish yellow, the surface is smooth and glossy, and the stripe pattern Have The meat is white, thick and dense. The folds are light grayish yellow, the density is dense, and the attachment to the fungus pattern is strong aspens. The fungus pattern is center or eccentric, the thickness is 3-4 cm, the length is 6-8 cm, the color is white, the shape is lower, the flesh is white, solid and very hard. The spores are oval or spindle-shaped, the surface is smooth, the size is 5-6 × 7-9 μm, and the spores are white.

以下、本発明を実施例により更に詳細に説明するが、本発明は、その趣旨を超えない限り、以下の実施例に限定されるものではない。   EXAMPLES Hereinafter, although an Example demonstrates this invention still in detail, this invention is not limited to a following example, unless the meaning is exceeded.

実施例１：
＜新規菌株の作出＞ Example 1:
<Creation of new strain>

＜新規菌株＞
子実体としては、株式会社キノックス所有のエリンギの子実体（Pleurotus eryngii、品種名ＫＸ−ＥＧ０７９号）と中国遼寧省瀋陽農業大学所有の白霊茸（Pleurotus nebrodensis）の子実体とを使用した。 <New strain>
As the fruiting body, the fruiting body of Elingi (Pleurotus eryngii, variety name KX-EG079) owned by Kinox Co., Ltd. and the fruiting body of Pleurotus nebrodensis owned by Shenyang University of Agriculture, Liaoning Province, China were used.

先ず、落下法により、エリンギ子実体と白霊茸の傘をペトリー皿に入れて静置し、得られた胞子紋に滅菌水を加えて胞子を懸濁させ、血球計算板を使用し、胞子懸濁液中の胞子数を計数後、適当な胞子濃度に希釈し、ＰＤＡ平板培地に接種し、２３℃で８日間培養した後、発芽した１次菌糸を分離し、各々の１次菌糸株を得た。次いで、ＰＤＡ平板培地に両１次菌糸株を接種し、２３℃で２０日間培養し、総当りの交配を行った。交配の有無は、光学顕微鏡下でクランプ結合を確認することにより行った。そして、エリンギ（Pleurotus eryngii）と白霊茸（Pleurotus nebrodensis）の交配株約５０株を得た。   First, put the eringi fruit body and the white mausoleum umbrella in a petri dish by the falling method, and leave the spore in sterilized water by adding sterile water to the obtained spore pattern. After counting the number of spores in the suspension, diluted to an appropriate spore concentration, inoculated on a PDA plate medium and cultured at 23 ° C. for 8 days, the germinated primary mycelium was separated, and each primary mycelium strain was isolated. Got. Subsequently, both primary mycelia strains were inoculated on a PDA plate medium, cultured at 23 ° C. for 20 days, and cross-bred. The presence or absence of mating was performed by confirming clamp binding under an optical microscope. As a result, about 50 crosses of Pleurotus eryngii and Pleurotus nebrodensis were obtained.

先ず、スギオガコとコーンコブミールを容積比で８：２の割合で混合し、培養基総重量当たり１０重量％の専管フスマを添加（１ビン当たり５２ｇ添加）した後、含水率を約６８重量％に調整した培養基に、上記の交配株約５０株を接種し、２３℃で４０日間培養することで、木粉培地に順応性の高い１０株の交配株を選抜した。   First, Sugiogako and corn cob meal are mixed at a volume ratio of 8: 2, and 10% by weight of special tubed bran is added to the total weight of the culture medium (52g added per bottle), and the water content is adjusted to about 68% by weight. About 50 strains of the above-mentioned hybrids were inoculated into the cultured medium and cultured at 23 ° C. for 40 days to select 10 crosses highly adaptable to the wood flour medium.

次いで、スギオガコとコーンコブミールを容積比で８：２の割合で混合し、培養基総重量当たり１２重量％のフスマを添加（１ビン当たり６０ｇ添加）した後、含水率を約６６重量％に調整した培養基に、上記の菌株１０菌株を接種し、２３℃で３５日間の培養管理を行った。培養の終了した菌床は、表面を約１５ｍｍの深さに菌掻き処理を行った後、倒立状態で環境温度１５〜１６℃、炭酸ガス濃度が８００〜２，０００ｐｐｍ、昼間の時間帯のみ４００Ｌｘ程度の光を照射して、環境湿度７０〜９８％の湿度範囲で芽出し管理を行った。   Next, Sugiogako and corn cob meal were mixed at a volume ratio of 8: 2, and 12% by weight of bran was added to the total weight of the culture medium (60 g per bottle), and the water content was adjusted to about 66% by weight. The culture medium was inoculated with 10 strains described above, and the culture was managed at 23 ° C. for 35 days. After the cultivation, the surface of the fungus bed is scraped to a depth of about 15 mm, and in an inverted state, the ambient temperature is 15 to 16 ° C., the carbon dioxide concentration is 800 to 2,000 ppm, and the daytime is 400 Lx only. The sprouting was controlled in a humidity range of 70 to 98% by irradiating a certain amount of light.

芽切り確認後は、正立状態に戻した後、環境温度を１６〜１８℃、環境湿度を８０〜９０％に変更して、芽出し管理と同様の照度環境下で生育管理を行い、優良交配株を５株選抜し、再度同培地にて同様の生育管理を５回繰り返し、最良な１菌株を育成した。この菌株は、「ＫＸ−ＥＮ２００１」と命名され「ＦＥＲＭ Ｐ−２０５７６」として寄託されている。   After confirming sprouting, after returning to an upright state, the environment temperature is changed to 16-18 ° C and the environmental humidity is changed to 80-90%. Five strains were selected and the same growth control was repeated five times in the same medium again to grow the best strain. This strain is named “KX-EN2001” and has been deposited as “FERM P-20576”.

＜新規子実体の栽培方法＞
先ず、スギオガコとコーンコブミールを容積比で８：２の割合に混合し、培養基総重量当たり３重量％の専かんフスマと同６重量％の乾燥オカラ、更に１重量％のネオビタスＮを添加（合計で１ビン当たり８０ｇ添加）した後、含水率を約６８重量％に調節して培養基を調製した。充填機を使用し、ポリプロピレン製栽培瓶（８５０cc）に正味重量で５３０〜５４０ｇの培養基を充填し、培養基の中央部に直径が約１５ｍｍで底部に到達する接種孔を設けて施栓した。次いで、常法に従って高圧殺菌釜で殺菌した後に冷却した。冷却は、放冷時における戻り空気による再汚染を防止するため、クリーンルーム内で行った。 <Cultivation method of new fruit body>
First, Sugiogako and corn cob meal are mixed at a volume ratio of 8: 2, and 3% by weight of the total weight of the culture medium, 6% by weight of dried okara, and 1% by weight of neovitus N are added (total) After adding 80 g per bottle, the water content was adjusted to about 68% by weight to prepare a culture medium. Using a filling machine, a polypropylene culture bottle (850 cc) was filled with a culture medium of 530 to 540 g in net weight, and an inoculation hole having a diameter of about 15 mm and reaching the bottom was provided at the center of the culture medium and plugged. Next, the mixture was sterilized in a high-pressure sterilization pot according to a conventional method and then cooled. Cooling was performed in a clean room to prevent recontamination due to return air during cooling.

その後、同クリーンルーム内で無菌的にＫＸ−ＥＮ２００１菌株を接種して培養を開始した。培養は２３℃で３５日間の培養管理を行った後、培地の表面も含めて約１５ｍｍの深さに菌掻き処理を行った。その後、倒立状態で直ちに、環境温度１５〜１６℃、炭酸ガス濃度が８００〜２，０００ｐｐｍ、昼間の時間帯のみ４００Ｌｘ程度の光を照射して、環境湿度７０〜９８％の湿度範囲で低湿度環境と高湿度環境とを一定間隔で保持しながら繰り返す、芽出し管理を行った。   Thereafter, the KX-EN2001 strain was aseptically inoculated in the same clean room and culture was started. Culturing was carried out at 23 ° C. for 35 days, and then the bacteria were scraped to a depth of about 15 mm including the surface of the medium. After that, immediately in an inverted state, the ambient temperature is 15 to 16 ° C., the carbon dioxide concentration is 800 to 2,000 ppm, and light of about 400 Lx is irradiated only during the daytime, and the humidity is low in the humidity range of 70 to 98%. The sprout management was repeated while maintaining the environment and the high humidity environment at regular intervals.

芽切り確認後は、容器を正立状態に戻した後、環境温度を１６〜１８℃、環境湿度を８０〜９０％に変更して、芽出し管理と同様の照度環境下で生育を行った。そして、きのこは菌傘が平らになるまで生長させた段階で、１本づつ収穫を行った。収穫までの日数は１４．３日（標準偏差値０．６）、１瓶当たりの収量は１３９．４ｇ（２１．７）、有効茎本数は３．１本（１．４）、子実体の個重は４５．０ｇ（９．４）であった。子実体の形態を図１に示す。   After sprouting was confirmed, the container was returned to an upright state, and then the environmental temperature was changed to 16 to 18 ° C. and the environmental humidity was changed to 80 to 90%. The mushrooms were harvested one by one when they were grown until the fungus was flat. The number of days until harvest is 14.3 days (standard deviation 0.6), the yield per bottle is 139.4 g (21.7), the number of effective stems is 3.1 (1.4), The individual weight was 45.0 g (9.4). The form of the fruiting body is shown in FIG.

比較例１：
実施例１において、使用する種菌の種類を白霊茸（Pleurotus nebrodensis）に変更した以外は、実施例１と同様の管理で白霊茸（Pleurotus nebrodensis）の栽培を行った。結果においては、実施例１とは異なり、子実体の発生をまったく認めることは出来なかった。 Comparative Example 1:
In Example 1, except that the type of inoculum used was changed to white leopard (Pleurotus nebrodensis), white leopard (Pleurotus nebrodensis) was cultivated under the same management as in Example 1. In the results, unlike Example 1, the occurrence of fruiting bodies could not be recognized at all.

上記の実施例１及び比較例１の結果から明らかな通り、本発明に従い、エリンギ（Pleurotus eryngii）と白霊茸（Pleurotus nebrodensis）との変種間きのこを交配させて作出した新規な菌株は、白霊茸（Pleurotus nebrodensis）栽培と異なり、一般的に使用されるエリンギ栽培用の人工培養基を使用して容易に人工栽培を行うことが出来る。   As is clear from the results of Example 1 and Comparative Example 1 above, according to the present invention, a novel strain produced by crossing mushrooms between varieties of Elingi (Pleurotus eryngii) and white leopard (Pleurotus nebrodensis) is white. Unlike mausoleum (Pleurotus nebrodensis) cultivation, it is possible to easily carry out artificial cultivation using a commonly used artificial culture medium for eringi cultivation.

実施例１で得られた新規子実体の形態説明図Form explanatory drawing of the new fruiting body obtained in Example 1

Claims (3)

エリンギの胞子と白霊茸の胞子とを交配させた後、交配菌を培養して選抜を行うことを特徴とする新規菌株の作出方法。   A method for producing a novel strain, comprising: mating a spore of eringi and a spore of a white mausoleum, and then culturing and selecting the hybrid. プレオロウタス属菌株であるＫＸ−ＥＮ２００１菌株（ＦＥＲＭ Ｐ−２０５７６）。   KX-EN2001 strain (FERM P-20576) which is a strain of the genus Pleurotus. 栽培容器にオガコと栄養源とを混合して水分調整した培養基を充填し、培養基を加熱殺菌した後、請求項１で得られた新規菌株の種菌を接種し、芽出室に搬入し、芽出しを行ない、次いで、原基の生育を行なうことを特徴とする新規子実体の栽培方法。   A cultivation container is filled with a culture medium mixed with sawdust and nutrients, and the moisture content is adjusted. The culture medium is sterilized by heating, then inoculated with the inoculum of the new strain obtained in claim 1, and carried into the budding chamber. And then growing the primordium.

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