JP3539953B2 - New strain of lyophilum ulmarium - Google Patents

Description

【０００９】
官能検査は、リオフィラム ウルマリウム子実体を油炒めし、硫酸キニーネを、苦味の標準物質として、パネラーが判定する。油炒めは次の様に行う。すなわち、得られたリオフィラム ウルマリウムの子実体の柄の基部を除去し、１本ずつにわけ、軽く水洗いして、よく水をきる。次いで、フライパンにサラダ油５ｍｌを加え、全体にのばし、中火で油を充分に熱する。前述リオフィラム ウルマリウム子実体１００ｇ（湿重）を加え、箸でかきまぜながら、２分間以上、全体に火を通し、フライパンに水気が無くなれば、火を止める。該子実体を常温になるまで冷まし、パネラーに苦くない、苦い、かなり苦い、著しく苦いの四段階で評価させる。なお、以下、各表中の−は苦くない、＋は苦い、＋＋はかなり苦い、＋＋＋は著しく苦いを意味する。この＋、−の中間がリオフィラム ウルマリウム子実体の苦味の硫酸キニーネを標準物質とした苦味の閾値、すなわち、ほとんど苦味を感じない苦味であり、標準物質の硫酸キニーネの閾値、０．０００００８Ｍ（太田静行著、「食品調味の知識」第２版第１刷、昭和６０年１０月１日、幸書房発行、第３８頁）の苦味に相当する。
前述の供試菌株中、リオフィラム ウルマリウム Ｌｕ １−２が最も低苦味菌株であるが、Ｂ、Ｃ、Ｄ培地を用いた場合は、収量は増加するものの得られる子実体は苦味のあるものとなる。
【００１０】
本発明において高苦味化培地とは、リオフィラム ウルマリウム Ｌｕ １−２の人工栽培を行った際に得られる子実体の苦味が官能検査により＋以上の培地を意味し、上記Ｂ、Ｃ、Ｄ培地である。また本発明の新菌株とは、該菌株を高苦味化培地で栽培して得られる子実体の苦味が、少なくとも一種の高苦味化培地について、苦味をほとんど感じない程度以下の菌株をいう。
【００１１】
次に、本発明の新菌株の育種について述べる。
１．選抜育種
自然界に発生しているリオフィラム ウルマリウム１１２個の子実体より組織分離を行い、純粋分離した菌糸体９０菌株を得た。次にこの９０菌株を前記Ｂ培地を用い栽培試験を行った。栽培試験により、子実体を形成した菌株４８株の子実体を官能検査に供した。子実体の形状に優れ、収量も多く、苦味の低い菌株１０株を選抜した。更にこの１０株をＢ、Ｃ、Ｄ培地で栽培試験し、工業的栽培に適し、最も低苦味な菌株として、リオフィラム ウルマリウム Ｌｕ １−１３を選抜した。
【００１２】
以下、このリオフィラム ウルマリウムについて説明する。
リオフィラム ウルマリウム Ｌｕ １−１３株は、福島県裏磐梯にて枯れ木に叢生していた子実体より本発明者らが組織分離したもので子実体及び胞子の特徴は次のようである。子実体は叢生、カサは径５〜１．５cm、円形又は不正形で丸山形、表面は平滑、湿潤、白色〜帯褐クリーム色を呈しており、往々やや濃色の斑紋を現し、老時中央にき裂を生じることがある。肉は白色、幅広く柄に上生する。柄は偏心性で湾曲し、３〜７×１〜２cm、カサとほぼ同色、頂部は白色で綿毛状ないし粉状である。胞子はほぼ球形、平滑、無色、４．５〜５．５×３．５〜４．５μｍ、紋は白色であった。
以上の特徴を伊藤誠哉著「日本菌類誌」第二巻第五号（１９５９年、養賢堂出版）の記載と比較すると、本菌はリオフィラム ウルマリウムであることが明りょうである。
【００１３】
次にリオフィラム ウルマリウム Ｌｕ １−１３株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
７日目：旺盛な生育。コロニー径は４０mm。白色で密な菌糸、気菌糸を多量に生じる。１０日目：シャーレ全体に菌糸が生育する。１７日目：表面全体に密な気菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
７日目：旺盛な生育。コロニー径は３６mm。白色で密な菌糸、気菌糸を多量に生じる。１０日目：シャーレ全体に菌糸が生育する。１７日目：表面全体に密な気菌糸を生じる。コロニー中央部は薄い黄色、他は白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
７日目：小程度の生育。コロニー径は２６mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。１７日目：シャーレ全体に菌糸が生育する。菌糸は樹状で希薄、白色。
（４）サブロー寒天培地（２５℃培養）
７日目：旺盛な生育。コロニー径は４２mm。白色で綿状の密な菌糸、気菌糸やや多い。１０日目：シャーレ全体に菌糸が生育する。気菌糸を極めて多量に生じ、菌糸は綿状で白色。
（５）オートミール寒天培地（２５℃培養）
７日目：旺盛な生育。コロニー径は３６mm。菌糸はよく分枝して伸び、気菌糸は少ない。１０日目：シャーレ全体に菌糸が生育する。綿状の気菌糸を多量に生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
７日目：小程度の生育。コロニー径は２１mm。菌糸は白色で直線的に伸長し、放射状のコロニーを形成する。１７日目：シャーレ全体に菌糸が生育する。気菌糸を多量に生じる。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
７日目：旺盛な生育。コロニー径は４０mm。白色で密な菌糸、気菌糸を多量に生じる。マット状。１０日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色だが、培地は黄色に変化する。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
７日目：小程度の生育。コロニー径は１８mm。菌糸は白色で短くマット状に生育、気菌糸は少ない。培地は褐変、褐変半径は４０mm。１７日目：中程度の生育。コロニー径は３７mm。褐変半径は４２mm。種菌の新旧により著しく生育速度に差が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは７〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００１４】
次に、リオフィラム ウルマリウム Ｌｕ １−１３株と他のリオフィラム
ウルマリウムとの異同判定として、両菌糸が持つ性因子が異なっていれば、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、性因子の異同を寒天培地上における対峙培養によって調べた。
供試したリオフィラム ウルマリウムとしてはリオフィラム ウルマリウム
ＩＦＯ ９６３７、リオフィラム ウルマリウム ＩＦＯ ３０５２５、リオフィラム ウルマリウム ＩＦＯ ３０７７５、リオフィラム ウルマリウム Ｌｕ １−２、リオフィラム ウルマリウム Ｌｕ １−８、リオフィラム ウルマリウム Ｌｕ １−１７、リオフィラム ウルマリウム Ｍ−８１７１、リオフィラム ウルマリウム ＳＡである。なおリオフィラム ウルマリウム ＳＡは、市販のリオフィラム ウルマリウム子実体よりの分離株である。
上記それぞれのリオフィラム ウルマリウムの二核菌糸を保存スラントより３×３×３mmのブロックとして切り出し、それぞれをＰＧＹ寒天培地の中央部に、リオフィラム ウルマリウム Ｌｕ １−１３の二核菌糸と対峙して植菌し（２cm間隔）、２５℃、１４日間培養後、両菌株のコロニー間に帯線が生じるか否かを判定した。結果を表２に示す。（帯線を生じた場合＋、生じなかった場合−）。
【００１５】
【表２】

【００１６】
表２よりわかるように、前記各菌株は、リオフィラム ウルマリウム Ｌｕ
１−１３との対峙培養ですべて帯線を生じ、このことからリオフィラム ウルマリウム Ｌｕ １−１３は新しい菌株であることは明白である。
以上説明したように本発明の選抜育種による新菌株として、例えばリオフィラム ウルマリウム Ｌｕ １−１３が挙げられるが、前記菌株と同様に高苦味化培地で栽培して得られる子実体の苦味が、少なくとも一種の高苦味化培地について、苦味をほとんど感じない程度以下であるという特性を示す菌株は、すべて本発明に属するものである。
【００１７】
２．交配育種
本発明の新菌株を交配育種により得た。
特開昭６３−２７３４６７号公報に記載のように、リオフィラム ウルマリウム Ｌｕ １−８とリオフィラム ウルマリウム Ｌｕ １−１７の交配により、収量、生育速度、子実体形態に優れたリオフィラム ウルマリウム Ｍ−８１７１が得られている。栽培特性に優れ、かつ子実体の苦味が低減された菌株を交配育種するために、このリオフィラム ウルマリウム Ｌｕ １−８、リオフィラム ウルマリウム Ｌｕ １−１７の両菌株を親株として用いた。
リオフィラム ウルマリウム Ｌｕ １−８とリオフィラム ウルマリウム
Ｌｕ １−１７をＡ培地にそれぞれ生育させ、通常の操作により子実体を発生させた。該子実体よりそれぞれ胞子を回収し、発芽させた一核菌糸を単離して、５７株ずつを総当り交配して１６５３株を得た。この１６５３株をＢ培地を用いて栽培試験を行い、官能検査により低苦味な菌株８０株を選択し、次にＢ、Ｃ、Ｄ培地で栽培試験を行い、栽培特性に優れ、苦味が、少なくとも一種の高苦味化培地について、苦味をほとんど感じない程度以下である８菌株を選抜し、それぞれリオフィラム ウルマリウム Ｋ−０４２９、リオフィラム ウルマリウム Ｋ−０２０７、リオフィラム ウルマリウム Ｋ−０２０２、リオフィラム ウルマリウム Ｋ−０２５９、リオフィラム ウルマリウム Ｋ−１００４、リオフィラム ウルマリウム Ｋ−１２５７、リオフィラム ウルマリウム Ｋ−６８０４、リオフィラム ウルマリウム Ｋ−６８０６とそれぞれ命名した。
【００１８】
次にこのリオフィラム ウルマリウム Ｋ−０４２９株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
５日目：コロニー径は１８mm。白色で密な菌糸、気菌糸を多量に生じる。１０日目：コロニー径は４７mm。２０日目：コロニー径は８３mm。放射状に生育して表面全体に密な気菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は４０mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：コロニー径は５５mm。２０日目：コロニー径は６６mm。表面全体に密な気菌糸を生じる。綿状のコロニーで、菌糸は白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は２８mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。２０日目：コロニー径は５２mm。菌糸は樹状で希薄、白色。
（４）サブロー寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は５０mm。白色で綿状の密な菌糸、気菌糸やや多い。２０日目：シャーレ全体に菌糸が生育する。放射状に生育して表面全体に気菌糸を生じる。菌糸は白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は５１mm。綿状のコロニーで、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。綿状のコロニーで、気菌糸を多量に生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は２７mm。菌糸は白色で、樹枝状のコロニーを形成する。２０日目：コロニー径は４８mm。気菌糸を生じる。樹枝状のコロニーで、菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は４９mm。綿状のコロニーで白色な菌糸、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
５日目：小程度の生育。コロニー径は１８mm。菌糸は白色で気菌糸を多量に生じる。培地は褐変して、褐変半径は３４mm。１５日目：旺盛な生育。コロニー径は６４mm。培地全面が褐変。種菌の新旧により著しく生育速度に差が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは７〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００１９】
次にリオフィラム ウルマリウム Ｋ−０２０７株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６１mm。白色で放射状のコロニー、気菌糸を少量生じる。２０日目：シャーレ全体に菌糸が生育する。気菌糸は少ない。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は５３mm。白色で密な菌糸、気菌糸を多量に生じる。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。コロニーは綿状で白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
７日目：旺盛な生育。コロニー径は５１mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。１５日目：コロニー径は７５mm。菌糸は樹状で希薄、白色。
２０日目：コロニー径は７７mm。
（４）サブロー寒天培地（２５℃培養）
１０日目：とても旺盛な生育。コロニー径は５８mm。白色で放射状の密な菌糸、気菌糸やや多い。１５日目：シャーレ全体に菌糸が生育する。気菌糸を生じ、菌糸は放射状で白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は６６mm。コロニーは綿状で、気菌糸を生じる。１５日目：シャーレ全体に菌糸が生育する。綿状のコロニーで気菌糸を生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は３３mm。菌糸は白色でやや薄く、樹枝状に伸長し、気菌糸を伴う。２０日目：コロニー径は７４mm。コロニーは希薄で樹枝状、菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は６５mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６３mm。菌糸は白色で、気菌糸を多量に生じる。培地は褐変、褐変半径は６５mm。１５日目：シャーレ全体に菌糸が生育する。気菌糸を多量に生じ、培地全面が褐変する。種菌の新旧により著しく生育速度に差が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは７〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２０】
次にリオフィラム ウルマリウム Ｋ−０２０２株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６０mm。白色で放射状のコロニー、気菌糸を少量生じる。２０日目：シャーレ全体に菌糸が生育する。気菌糸は少ない。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は５９mm。白色で密な菌糸、気菌糸を多量に生じる。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。コロニーは綿状で白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
５日目：旺盛な生育。コロニー径は１６mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。１５日目：コロニー径は６２mm。菌糸は樹状で希薄、白色。
２０日目：コロニー径は７７mm。
（４）サブロー寒天培地（２５℃培養）
１０日目：とても旺盛な生育。コロニー径は６６mm。白色で放射状の密な菌糸、気菌糸やや多い。１５日目：シャーレ全体に菌糸が生育する。気菌糸を生じ、菌糸は放射状で白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は６８mm。コロニーは綿状で、気菌糸を生じる。１５日目：シャーレ全体に菌糸が生育する。綿状のコロニーで気菌糸を生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は３３mm。菌糸は白色でやや薄く、樹枝状に伸長し、気菌糸を伴う。２０日目：コロニー径は６２mm。コロニーは希薄で樹枝状、菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は６８mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６１mm。菌糸は白色で、気菌糸を多量に生じる。培地は褐変。１５日目：シャーレ全体に菌糸が生育する。気菌糸を多量に生じ、培地全面が褐変する。種菌の新旧により著しく生育速度に差が生じる。（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは７〜８．５であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２１】
次にこのリオフィラム ウルマリウム Ｋ−０２５９株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
５日目：コロニー径は２４mm。白色で密な菌糸、気菌糸を多量生じる。１０日目：コロニー径は６４mm。１５日目：シャーレ全体に菌糸が生育する。放射状に生育して表面全体に密な気菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は６１mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：コロニー径は８５mm。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。綿状のコロニーで、菌糸は白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は４８mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。２０日目：コロニー径は７５mm。菌糸は樹状で希薄、白色。
（４）サブロー寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６５mm。白色で綿状の密な菌糸、気菌糸やや多い。２０日目：シャーレ全体に菌糸が生育する。放射状に生育して表面全体に気菌糸を生じる。菌糸は白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６７mm。綿状のコロニーで、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。綿状のコロニーで、気菌糸を多量に生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は３９mm。菌糸は白色で、樹枝状のコロニーを形成する。２０日目：コロニー径は６６mm。気菌糸を生じる。樹枝状のコロニーで、菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６３mm。綿状のコロニーで白色な菌糸、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
５日目：小程度の生育。コロニー径は２９mm。菌糸は白色で気菌糸を多量に生じる。培地は褐変。１５日目：旺盛な生育。コロニー径は８８mm。培地全面が褐変。種菌の新旧により著しく生育速度に差が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは６〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２２】
次にリオフィラム ウルマリウム Ｋ−１００４株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６４mm。白色で放射状のコロニー、気菌糸を少量生じる。２０日目：シャーレ全体に菌糸が生育する。気菌糸は少ない。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６４mm。白色で密な菌糸、気菌糸を多量に生じる。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。コロニーは綿状で白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
５日目：旺盛な生育。コロニー径は１４mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。１５日目：コロニー径は６０mm。菌糸は樹状で希薄、白色。２０日目：コロニー径は７５mm。
（４）サブロー寒天培地（２５℃培養）
１０日目：とても旺盛な生育。コロニー径は８３mm。白色で放射状の密な菌糸、気菌糸やや多い。１５日目：シャーレ全体に菌糸が生育する。気菌糸を生じ、菌糸は放射状で白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は８１mm。コロニーは綿状で、多量の気菌糸を生じる。１５日目：シャーレ全体に菌糸が生育する。綿状のコロニーで気菌糸を生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は２３mm。菌糸は白色でやや薄く、樹枝状に伸張し、気菌糸を伴う。２０日目：コロニー径は４５mm。コロニーは希薄で樹枝状。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は８０mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６８mm。菌糸は白色で、気菌糸を多量に生じる。培地は褐変。１５日目：シャーレ全体に菌糸が生育する。気菌糸を多量に生じ、培地全面が褐変する。種菌の新旧により著しく生育速度に差が生じる。（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは６〜７であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２３】
次にリオフィラム ウルマリウム Ｋ−１２５７株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７１mm。白色で放射状のコロニー、気菌糸を少量生じる。２０日目：シャーレ全体に菌糸が生育する。気菌糸は少ない。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６６mm。白色で密な菌糸、気菌糸を多量に生じる。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。コロニーは綿状で白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
５日目：旺盛な生育。コロニー径は２６mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。１５日目：コロニー径は７１mm。菌糸は樹状で希薄、白色。２０日目：コロニー径は８１mm。
（４）サブロー寒天培地（２５℃培養）
１０日目：とても旺盛な生育。コロニー径は７９mm。白色で放射状の密な菌糸、気菌糸やや多い。１５日目：シャーレ全体に菌糸が生育する。気菌糸を生じ、菌糸は放射状で白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は７９mm。コロニーは綿状で、多量の気菌糸を生じる。１５日目：シャーレ全体に菌糸が生育する。綿状のコロニーで気菌糸を生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は３７mm。菌糸は白色でやや薄く、樹枝状に伸長し、気菌糸を伴う。２０日目：コロニー径は６８mm。コロニーは希薄で樹枝状。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：極めて旺盛な生育。コロニー径は６４mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７０mm。菌糸は白色で、気菌糸を多量に生じる。培地は褐変。１５日目：シャーレ全体に菌糸が生育する。気菌糸を多量に生じ、培地全面が褐変する。種菌の新旧により著しく生育速度に差が生じる。（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは６〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２４】
次にこのリオフィラム ウルマリウム Ｋ−６８０４株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
５日目：コロニー径は２５mm。白色で密な菌糸、気菌糸を多量に生じる。１０日目：コロニー径は６１mm。１５日目：コロニー径は８３mm。放射状に生育して表面全体に密な気菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は６１mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：コロニー径は８６mm。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。綿状のコロニーで、菌糸は白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は４９mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。２０日目：コロニー径は７６mm。菌糸は樹状で希薄、白色。
（４）サブロー寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７０mm。白色で綿状の密な菌糸、気菌糸やや多い。２０日目：シャーレ全体に菌糸が生育する。放射状に生育して表面全体に気菌糸を生じる。菌糸は白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７４mm。綿状のコロニーで、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。綿状のコロニーで、気菌糸を多量に生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は２９mm。菌糸は白色で、樹枝状のコロニーを形成する。２０日目：コロニー径は５３mm。気菌糸を生じる。樹枝状のコロニーで、菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７２mm。綿状のコロニーで白色な菌糸、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
５日目：小程度の生育。コロニー径は２８mm。菌糸は白色で気菌糸を多量に生じる。培地は褐変。１５日目：シャーレ全体に菌糸が生育する。培地全面が褐変。種菌の新旧により著しく生育速度に差が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは６〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２５】
次にこのリオフィラム ウルマリウム Ｋ−６８０６株の菌学的諸性質を示す。
（１）麦芽エキス寒天培地（２５℃培養）
５日目：コロニー径は１９mm。白色で密な菌糸、気菌糸を多量生じる。１０日目：コロニー径は５９mm。１５日目：コロニー径は８１mm。放射状に生育して表面全体に密な気菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃培養）
１０日目：中程度の生育。コロニー径は５５mm。白色で密な菌糸、気菌糸を多量に生じる。１５日目：コロニー径は８０mm。２０日目：シャーレ全体に菌糸が生育する。表面全体に密な気菌糸を生じる。綿状のコロニーで、菌糸は白色。
（３）ツァペック・ドックス寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は３８mm。樹状に伸長し極めて希薄な菌糸、気菌糸は少ない。２０日目：コロニー径は６６mm。菌糸は樹状で希薄、白色。
（４）サブロー寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は６６mm。白色で綿状の密な菌糸、気菌糸やや多い。２０日目：シャーレ全体に菌糸が生育する。放射状に生育し表面全体に気菌糸を生じる。菌糸は白色。
（５）オートミール寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７１mm。綿状のコロニーで、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。綿状のコロニーで、気菌糸を多量に生じる。菌糸は白色。
（６）合成ムコール寒天培地（２５℃培養）
１０日目：小程度の生育。コロニー径は３１mm。菌糸は白色で、樹枝状のコロニーを形成する。２０日目：コロニー径は６１mm。気菌糸を生じる。樹枝状のコロニーで、菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃培養）
１０日目：旺盛な生育。コロニー径は７０mm。綿状のコロニーで白色な菌糸、気菌糸を生じる。２０日目：シャーレ全体に菌糸が生育する。密な気菌糸を多量に生じる。菌糸は白色。
（８）フェノールオキシダーゼ検定用培地（２５℃培養）
５日目：小程度の生育。コロニー径は２８mm。菌糸は白色で気菌糸を多量に生じる。培地は褐変。１５日目：旺盛な生育。コロニー径は８６mm。培地全面が褐変。種菌の新旧により著しく生育速度に差が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径５mmの円盤状種菌を接種し、各温度で１２日間培養した後、コロニー径を測定した。その結果、最適な成育温度は２５℃付近であった。また、５℃、３５℃ではほとんど生育しなかった。
(10) 最適生育ｐＨ
ＰＧＹ液体培地（寒天を含まないＰＧＹ寒天培地）６０mlずつを１００ml容三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に種菌を接種して、２５℃、１５日間静置培養した後、菌体の乾燥重量を測定した。その結果、最適生育ｐＨは６〜８であった。また、生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
【００２６】
次に、リオフィラム ウルマリウム Ｋ−０４２９株、リオフィラム ウルマリウム Ｋ−０２０７株、リオフィラム ウルマリウム Ｋ−０２０２株、リオフィラム ウルマリウム Ｋ−０２５９株、リオフィラム ウルマリウム Ｋ−１００４株、リオフィラム ウルマリウム Ｋ−１２５７株、リオフィラム ウルマリウム Ｋ−６８０４株及びリオフィラム ウルマリウム Ｋ−６８０６株と他のリオフィラム ウルマリウムとの異同判定として、両菌糸が持つ性因子が異なっていれば、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、性因子の異同を寒天培地上における対峙培養によって調べた。
供試したリオフィラム ウルマリウムとしてはリオフィラム ウルマリウム
ＩＦＯ ９６３７、リオフィラム ウルマリウム ＩＦＯ ３０５２５、リオフィラム ウルマリウム ＩＦＯ ３０７７５、リオフィラム ウルマリウム Ｌｕ １−２、リオフィラム ウルマリウム Ｌｕ １−８、リオフィラム ウルマリウム Ｌｕ １−１７、リオフィラム ウルマリウム Ｍ−８１７１、リオフィラム ウルマリウム ＳＡ、リオフィラム ウルマリウム Ｌｕ １−１３である。
【００２７】
上記それぞれのリオフィラム ウルマリウムの二核菌糸を保存スラントより３×３×３mmのブロックとして切り出し、それぞれをＰＧＹ寒天培地の中央部に、リオフィラム ウルマリウム Ｋ−０４２９株、リオフィラム ウルマリウム
Ｋ−０２０７株、リオフィラム ウルマリウム Ｋ−０２０２株、リオフィラムウルマリウム Ｋ−０２５９株、リオフィラム ウルマリウム Ｋ−１００４株、リオフィラム ウルマリウム Ｋ−１２５７株、リオフィラム ウルマリウム Ｋ−６８０４株、又はリオフィラム ウルマリウム Ｋ−６８０６株の二核菌糸と対峙して植菌し（２cm間隔）、２５℃、１４日間培養後、両菌株のコロニー間に帯線が生じるか否かを判定した。結果を表３、表４に示す。（帯線を生じた場合＋、生じなかった場合−）。
【００２８】
【表３】

【００２９】
【表４】

【００３０】
表３、表４よりわかるようにリオフィラム ウルマリウム Ｋ−０４２９、リオフィラム ウルマリウム Ｋ−０２０７、リオフィラム ウルマリウム Ｋ−０２０２、リオフィラム ウルマリウム Ｋ−０２５９、リオフィラム ウルマリウム Ｋ−１００４、リオフィラム ウルマリウム Ｋ−１２５７、リオフィラム ウルマリウム Ｋ−６８０４、及びリオフィラム ウルマリウム Ｋ−６８０６はそれぞれ他の菌株との対峙培養ですべて帯線を生じ、このことから両菌株は新しい菌株であることは明白である。
【００３１】
以上説明したように本発明の交配により育種した新菌株として、例えばリオフィラム ウルマリウム Ｋ−０４２９、リオフィラム ウルマリウム Ｋ−０２０７、リオフィラム ウルマリウム Ｋ−０２０２、リオフィラム ウルマリウム Ｋ−０２５９、リオフィラム ウルマリウム Ｋ−１００４、リオフィラムウルマリウム Ｋ−１２５７、リオフィラム ウルマリウム Ｋ−６８０４、リオフィラム ウルマリウム Ｋ−６８０６等が挙げられるが、前記菌株と同様に高苦味化培地で栽培して得られる子実体の苦味が、少なくとも一種の高苦味化培地について、苦味をほとんど感じない程度以下であるという特性を示す菌株はすべて本発明に属するものである。
【００３２】
本発明による新菌株は、前述のとおり人工栽培時において高苦味化培地で栽培しても子実体の苦味が、少なくとも一種の高苦味化培地について、苦味をほとんど感じない程度以下であるという特性をもち、培地の種類に影響されず安定して子実体の苦味が少ない。本発明の新菌株のＡ、Ｂ、Ｃ、Ｄ培地を用い人工栽培を行い得た収穫適期の子実体の苦味、及び収量を表５、表６に示す。なお、前出リオフィラム ウルマリウム ＳＡの結果も合せ表６中に示す。
【００３３】
【表５】

【００３４】
【表６】

【００３５】
表５、表６に示すようにリオフィラム ウルマリウム Ｌｕ １−１３、リオフィラム ウルマリウム Ｋ−０４２９、リオフィラム ウルマリウム Ｋ−０２０７、リオフィラム ウルマリウム Ｋ−０２０２、リオフィラム ウルマリウム Ｋ−０２５９、リオフィラム ウルマリウム Ｋ−１００４、リオフィラム ウルマリウム Ｋ−１２５７、リオフィラム ウルマリウム Ｋ−６８０４、リオフィラム ウルマリウム Ｋ−６８０６の各菌株は、従来の人工栽培可能な菌株において、収穫適期の子実体が苦味を呈する培地を使用しても苦味を示さず、子実体収量を増加させるが苦味も増加させるので従来使用することが困難であった高苦味化培地を用いても、苦味の無い子実体を高収率で安定して得ることができる。
【００３６】
また、上記菌株を培地に接種して生成する菌糸体も苦味を呈さず、該菌糸体を食品に使用するにも好適である。なお、リオフィラム ウルマリウム菌糸体は食物繊維も多く、制ガン作用も知られており、該菌糸体を利用した食品は、健康維持のために特に有用である。
【００３７】
本発明において育種した新菌株のリオフィラム ウルマリウム Ｌｕ １−１３、リオフィラム ウルマリウム Ｋ−０４２９、リオフィラム ウルマリウムＫ−０２０７、リオフィラム ウルマリウム Ｋ−０２０２、リオフィラム ウルマリウム Ｋ−０２５９、リオフィラム ウルマリウム Ｋ−１００４、リオフィラム ウルマリウム Ｋ−１２５７、リオフィラム ウルマリウム Ｋ−６８０４、リオフィラム ウルマリウム Ｋ−６８０６はそれぞれ Lyophyllum ulmarium Ｌｕ １−１３、 Lyophyllum ulmarium Ｋ−０４２９、 Lyophyllum ulmarium Ｋ−０２０７、 Lyophyllum ulmarium Ｋ−０２０２、 Lyophyllum ulmarium Ｋ−０２５９、 Lyophyllum ulmarium Ｋ−１００４、 Lyophyllum ulmarium Ｋ−１２５７、 Lyophyllum ulmarium Ｋ−６８０４、 Lyophyllum ulmarium Ｋ−６８０６と表示され、工業技術院微生物工業技術研究所にそれぞれ微工研菌寄第１２５７１号（ＦＥＲＭＰ−１２５７１）、微工研菌寄第１２５７０号（ＦＥＲＭ Ｐ−１２５７０）、微工研菌寄第１２５６９号（ＦＥＲＭ
Ｐ−１２５６９）、微工研菌寄第１２９８０号（ＦＥＲＭ Ｐ−１２９８０）、微工研菌寄第１２９８１号（ＦＥＲＭ Ｐ−１２９８１）、微工研菌寄第１２９８２号（ＦＥＲＭ Ｐ−１２９８２）、微工研菌寄第１２９８３号（ＦＥＲＭ
Ｐ−１２９８３）、微工研菌寄第１２９８４号（ＦＥＲＭ Ｐ−１２９８４）、微工研菌寄第１２９８５号（ＦＥＲＭ Ｐ−１２９８５）として寄託されている。
【００３８】
なお、リオフィラム ウルマリウム子実体中の苦味成分は次のように定量することもできる。
リオフィラム ウルマリウム子実体中の可食部分（カサ部、柄部）を凍結乾燥し、これを粉砕して子実体凍乾粉末を得る。この凍乾粉末０．５ｇ（乾重）を３５mlの酢酸エチルで２５℃、２４時間振とう抽出し、ろ過、減圧濃縮後１mlのメタノールに加温しながら溶解し、抽出サンプルとする。
前述のサンプル２００μｌより、ＦＰＬＣ〔ファルマシア ファイン ケミカルス（ Pharmacia Fine Chemicals ) 製〕を用いて、次の様に苦味成分画分を分取する。カラムは、Lober ＲＰ−８〔メルク（ Merck )製、φ１．０×２４cm〕を使用して、溶媒８５％メタノール（関東化学；液クロ用）、１．０ml／分、常温、検出波長２１０nmの条件下で、溶出時間１０〜４５分の画分を分取する。
分取画分は再び減圧濃縮し、メタノール：エタノール：水＝３：４：４の溶媒１．０mlに溶解して、その５０μｌをＨＰＬＣ〔（株）島津製作所製ＬＣ−６Ａシステム〕にて苦味成分画分を分取する。カラムは、μ Bondapak Ｃ₁₈〔ウォータース（ Waters ) 製、φ０．３９×３０cm〕を使用して、メタノール：エタノール：水＝３：４：４の溶媒（関東化学；液クロ用メタノール、ナカライテスク；特級エタノール）、０．７ml／分、温度４０℃、検出波長ＵＶ２１０nmの条件下で、溶出時間３２．５〜４４．０分の画分の乾燥重量を測定する。この画分に溶出する成分（以下画分Ａと略す）は、官能検査において強い苦味を呈し、リオフィラム ウルマリウム子実体の苦味は、この画分Ａに起因する。
【００３９】
また、本発明で培地に使用した綿実殻は、安価に入手でき、しかもきのこの収量を著しく増加させる、きのこの人工栽培に有用な基材として、本発明者らが見出したものである。この綿実殻を培地に使用する際は、他の培地基材と混合して使用しても良いし、単独で使用しても良い。綿実殻は培地の乾燥重量に対して１５〜６０％で使用した場合が最も増収効果が良いが、使用量はこれに限定されるものではない。この綿実殻を培地に使用し、収量よく栽培できるきのことしては、例えば本発明のリオフィラム ウルマリウムの他、シイタケ、ヒラタケ、エノキタケ、マイタケ等があり、例えばこれらのきのこの菌株を使用し、通常の人工栽培を行えば良い。
【００４０】
【実施例】
以下に本発明によるリオフィラム ウルマリウム新菌株の人工栽培実施例を示すが、本発明は以下の実施例の範囲にのみ限定されるものではない。
なお、下記の実施例１〜実施例３は、本発明の参考例として挙示した例である。
【００４１】
実施例１
液体ＰＧＹ培地（寒天を含まないＰＧＹ寒天培地）１００mlに、リオフィラムウルマリウム Ｌｕ １−１３（ＦＥＲＭ Ｐ−１２５７１）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３５日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１６８ｇで、官能検査において苦味は感じられなかった。
【００４２】
なお、子実体の官能検査は、次の様に行った。上記ビン栽培で得られた子実体の柄の基部を除去し、１本ずつに分け、軽く水洗いして、よく水を切った。一方、フライパンにサラダ油５mlを加え、全体にのばし、中火で油を充分に熱した後、子実体１００ｇ（湿重）を加え、箸でかきまぜながら２分間以上、全体に火を通し、フライパンに水気が無くなるまで炒め、火を止めた。油炒めした子実体は常温になるまで冷まし、硫酸キニーネを標準物質とし、１０名のパネラーで苦味を判定した。
【００４３】
実施例２
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｌｕ １−１３の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１４３ｇで、子実体の官能検査において苦味は感じられなかった。
【００４４】
実施例３
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｌｕ １−１３の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３７日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１５７ｇで、子実体の官能検査において苦味は感じられなかった。
【００４５】
実施例４
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−０４２９（ＦＥＲＭ Ｐ−１２５７０）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３２日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４３日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１８４ｇで、子実体の官能検査において苦味は感じられなかった。
【００４６】
実施例５
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０４２９の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を２８日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３７日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１５７ｇで、子実体の官能検査において苦味は感じられなかった。
【００４７】
実施例６
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０４２９の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を２８日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３７日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１６４ｇで、子実体の官能検査において苦味は感じられなかった。
【００４８】
実施例７
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−０２０７（ＦＥＲＭ Ｐ−１２５６９）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を４０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１０日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１４日間培養を続けて子実体を得た。
得られた子実体の収量は１８８ｇで、子実体の官能検査において苦味は感じられなかった。
【００４９】
実施例８
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０２０７の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３４日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３１日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１４日間培養を続けて子実体を得た。
得られた子実体の収量は１６１ｇで、子実体の官能検査において苦味は感じられなかった。
【００５０】
実施例９
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０２０７の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３３日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３２日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１１日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１７８ｇで、子実体の官能検査において苦味は感じられなかった。
【００５１】
実施例１０
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−０２０２（ＦＥＲＭ Ｐ−１２９８０）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を４５日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で８日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１８７ｇで、子実体の官能検査において苦味は感じられなかった。
【００５２】
実施例１１
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０２０２の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を４０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で８日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１２日間培養を続けて子実体を得た。
得られた子実体の収量は１５４ｇで、子実体の官能検査において苦味は感じられなかった。
【００５３】
実施例１２
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０２０２の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３６日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４４日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で８日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１６２ｇで、子実体の官能検査において苦味は感じられなかった。
【００５４】
実施例１３
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−０２５９（ＦＥＲＭ Ｐ−１２９８１）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を４５日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で８日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１２日間培養を続けて子実体を得た。
得られた子実体の収量は１９２ｇで、子実体の官能検査において苦味は感じられなかった。
【００５５】
実施例１４
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０２５９の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３５日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で８日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１６２ｇで、子実体の官能検査において苦味は感じられなかった。
【００５６】
実施例１５
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−０２５９の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３７日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４３日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で８日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１６５ｇで、子実体の官能検査において苦味は感じられなかった。
【００５７】
実施例１６
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−１００４（ＦＥＲＭ Ｐ−１２９８２）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１１日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１５日間培養を続けて子実体を得た。
得られた子実体の収量は１８８ｇで、子実体の官能検査において苦味は感じられなかった。
【００５８】
実施例１７
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−１００４の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３２日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４８日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１０日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１５日間培養を続けて子実体を得た。
得られた子実体の収量は１５８ｇで、子実体の官能検査において苦味は感じられなかった。
【００５９】
実施例１８
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−１００４の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を２８日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５２日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１１日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１６日間培養を続けて子実体を得た。
得られた子実体の収量は１６１ｇで、子実体の官能検査において苦味は感じられなかった。
【００６０】
実施例１９
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−１２５７（ＦＥＲＭ Ｐ−１２９８３）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１１日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１５日間培養を続けて子実体を得た。
得られた子実体の収量は１８６ｇで、子実体の官能検査において苦味は感じられなかった。
【００６１】
実施例２０
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−１２５７の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３２日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４８日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１６日間培養を続けて子実体を得た。
得られた子実体の収量は１６０ｇで、子実体の官能検査において苦味は感じられなかった。
【００６２】
実施例２１
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−１２５７の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を２８日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５２日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１０日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１４日間培養を続けて子実体を得た。
得られた子実体の収量は１５７ｇで、子実体の官能検査において苦味は感じられなかった。
【００６３】
実施例２２
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−６８０４（ＦＥＲＭ Ｐ−１２９８４）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１０日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１６日間培養を続けて子実体を得た。
得られた子実体の収量は１９５ｇで、子実体の官能検査において苦味は感じられなかった。
【００６４】
実施例２３
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−６８０４の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を２８日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３２日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１０日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１５日間培養を続けて子実体を得た。
得られた子実体の収量は１６３ｇで、子実体の官能検査において苦味は感じられなかった。
【００６５】
実施例２４
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−６８０４の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を２８日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３２日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１６日間培養を続けて子実体を得た。
得られた子実体の収量は１７１ｇで、子実体の官能検査において苦味は感じられなかった。
【００６６】
実施例２５
液体ＰＧＹ培地１００mlに、リオフィラム ウルマリウム Ｋ−６８０６（ＦＥＲＭ Ｐ−１２９８５）を接種し、２５℃で１０日間培養して液体種菌を得た。一方、コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後に前記液体種菌２０mlを接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３２日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３８日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１５日間培養を続けて子実体を得た。
得られた子実体の収量は１９１ｇで、子実体の官能検査において苦味は感じられなかった。
【００６７】
実施例２６
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−６８０６の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で４０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１５日間培養を続けて子実体を得た。
得られた子実体の収量は１６２ｇで、子実体の官能検査において苦味は感じられなかった。
【００６８】
実施例２７
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｋ−６８０６の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３１日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で３９日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１４日間培養を続けて子実体を得た。
得られた子実体の収量は１６５ｇで、子実体の官能検査において苦味は感じられなかった。
【００６９】
対照例１
コーンコブミール１３０ｇ、マメカワ６０ｇ、フスマ３０ｇをよく混合して水道水により水分含有率を６２％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｌｕ １−２の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３５日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５０日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１７８ｇで、子実体の官能検査において苦味が感じられた。
【００７０】
対照例２
針葉樹オガクズ５０ｇ、綿実殻粉砕物５０ｇ、米糠１００ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｌｕ １−２の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で９日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１４日間培養を続けて子実体を得た。
得られた子実体の収量は１４０ｇで、子実体の官能検査において苦味が感じられた。
【００７１】
対照例３
針葉樹オガクズ１００ｇ、マメカワ７０ｇをよく混合して水道水により水分含有率を６３％に調製したものを、ポリプロピレン製８５０ml容広口ビンに圧詰めして、ビン口部中央より下方に向かい直径１cmの穴を開けた後、該培養基を１２０℃で６０分間高圧蒸気滅菌して、常温まで冷却後にリオフィラム ウルマリウム Ｌｕ １−２の固体種菌を接種した。
暗所、２５℃、湿度５０〜６０％の条件下で該培養基を３０日間培養すると、ビン全体に菌糸がまん延した。更に、同条件下で５５日間培養を続けて子実体発生基を得た。該子実体発生基の上部菌糸層１cmを除去し、水道水２０mlを加え充分に給水させた後に余剰の水道水を捨て、１５℃、湿度９０〜９５％、照度２０ルックスの条件下で１０日間培養を続けて子実体原基を得て、更に照度を２００ルックスに上げて１３日間培養を続けて子実体を得た。
得られた子実体の収量は１５３ｇで、子実体の官能検査において苦味が感じられた。
【００７２】
【発明の効果】
以上説明した様に、本発明によれば、従来使用することができなかった高苦味化培地を用い栽培しても、子実体の苦味が低減したリオフィラム ウルマリウム子実体を得ることが可能となり、工業的に高収量で良質なリオフィラム ウルマリウム子実体及び菌糸体を提供することができる。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a new strain of basidiomycete, and more particularly to a new strain of Lyophyllum ulmarium.
[0002]
[Prior art]
In the natural world, liophyllum ulmarium is clustered or incarcerated in the dead trees of various hardwoods in the autumn, and has been eaten without taste because of its well-formed and crisp meat quality. In addition, in recent years, using a culture medium in which rice bran and other nutrients are mixed with sawdust, a fungus bed artificial cultivation method of cultivating in a bottle or a box has been established, and it is possible to stably harvest mushrooms throughout the year regardless of the season. It has become.
Briefly described here is the artificial cultivation of the bacterium of lyophilum ulmarium in a bottle or a box filled with a culture medium and steam-sterilized, inoculated with an inoculum in which the hypha was propagated in the same culture medium in advance, and cultured for a predetermined number of days. After that, a generating operation is performed, and fruiting bodies are obtained and harvested, thereby completing one cycle. Several strains are commercially available or provided through technical tie-ups as strains used for cultivation, and there are various forms and cultivation characteristics depending on the strain used (for example, see Patent Document 1).
[0003]
[Patent Document 1]
JP-A-63-273467
[0004]
[Problems to be solved by the invention]
However, all of the currently used lyophilum ulmarium strains contain bitter components in the fruiting bodies at harvest, and have a bitterness that makes them unedible on high bittering media that increase fruiting body yield. In addition, the bitter component varies depending on the type of the medium, and the bitterness varies greatly. In general, mushrooms are mainly used as vegetables in which the taste of the material is emphasized, so that the inclusion of a large amount of bitter components significantly lowers the commercial value. For this reason, even in the case of a medium substrate that significantly increases the yield of fruiting bodies, a medium that simultaneously increases the bitter component, that is, a highly bitter medium, cannot be used.
In view of the above-mentioned current situation, an object of the present invention is to provide a new strain having a bitterness component in a fruit body at the time of harvest that hardly feels bitterness even when cultivated in these highly bittering media. It is in.
[0005]
[Means for Solving the Problems]
The present invention provides a fruiting body obtained by cultivation in a highly bittering medium, wherein the bitterness of at least one kind of bittering medium is less bitter than 0.000008M quinine sulfate, and from the following strains or mutants thereof: The present invention relates to a new lyophilum ulmarium strain, which is a new lyophilum ulmarium strain to be selected. Lyophilum ulmarium K-0429 (FERM P-12570),
Lyophilum ulmarium K-0207 (FERM P-12569),
Lyophilum ulmarium K-0202 (FERM P-12980),
Lyophilum ulmarium K-0259 (FERM P-12981),
Lyophilum ulmarium K-1004 (FERM P-12982),
Lyophilum ulmarium K-1257 (FERM P-12983),
Lyophilum ulmarium K-6804 (FERM P-12984),
Liophyllum ulmarium K-6806 (FERM P-12985)
[0006]
BEST MODE FOR CARRYING OUT THE INVENTION
The present inventors conducted a sensory test of the fruit body of lyophilum ulmarium, clarified the relationship between the bitterness of the fruit body and the cultivation medium, and performed an artificial cultivation using a medium that increases the bitterness of the fruit body, that is, a high-bittering medium. In addition, the present inventors succeeded in breeding a new strain of lyophilum ulmarium in which the obtained fruiting body has a bitterness of at least one kind of highly bittering medium or less, at which bitterness is hardly felt, thereby completing the present invention.
[0007]
Hereinafter, the present invention will be described in detail.
As lyophilum ulmarium strains, lyophilum ulmarium Lu 1-2 (FERM P-12584), lyophilum ulmarium Lu 1-8 (FERM BP-1416), and lyophilum ulmarium Lu 1-17 (FERM) described in JP-A-63-273467. BP-1417) and lyophilum ulmarium M-8171 (FERM BP-1415), and artificially cultivating each strain according to the method described in the publication, harvesting fruiting bodies at the appropriate time for harvesting, and sensory testing of fruiting bodies Was done. As a culture medium for artificial cultivation, 50 g of softwood sawdust, 50 g of hardwood sawdust, and 90 g of rice bran described in the publication were mixed well and adjusted to a water content of 65% with tap water. After filling, a hole having a diameter of 1 cm was directed downward from the center of the bottle mouth, and then the sawdust solid culture medium stoppered with a cap was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes to obtain an A medium. In addition, according to the method for preparing the medium, 130 g of corn cob meal, 60 g of Mamekawa, and 30 g of bran were mixed well and similarly prepared, and B medium, 50 g of conifer sawdust, 50 g of crushed cotton hulls, and 100 g of rice bran were mixed well. The medium prepared above was mixed well with C medium, 100 g of coniferous sawdust, and 70 g of Mamekawa, and similarly prepared, and used as D medium.
Table 1 shows the yield (g / bin) of the fruiting bodies obtained in each used medium and the results of the sensory test.
[0008]
[Table 1]

[0009]
The sensory test is carried out by sautéing lipophilum ulmarium fruit bodies with oil, and panelists judge quinine sulfate as a reference substance for bitterness. Oil stir-fry is performed as follows. That is, the base of the stem of the obtained fruit body of lyophilum ulmarium is removed, divided into single pieces, lightly washed with water, and well drained. Next, add 5 ml of salad oil to a frying pan, spread the whole, and heat the oil sufficiently over medium heat. Add 100 g (wet weight) of the above-mentioned lyophilum ulmarium fruit body, stir with chopsticks and cook for at least 2 minutes. When the frying pan is dry, turn off the fire. The fruiting bodies are cooled to room temperature and evaluated by the panelists on a four-point scale: bitter, bitter, fairly bitter, and markedly bitter. In the following,-in each table means not bitter, + means bitter, ++ means quite bitter, and +++ means extremely bitter. The threshold between + and-is the bitterness threshold of lyophilum ulmarium fruiting body with bitter quinine sulfate as a standard substance, that is, the bitterness that hardly feels bitterness, and the threshold of the standard substance quinine sulfate, 0.000008M (Shizuki Ota) This book is equivalent to the bitterness of the book, "Knowledge of Food Seasoning", 2nd edition, 1st printing, published October 1, 1985, published by Koshobo, p. 38).
Among the test strains described above, lyophilum ulmarium Lu 1-2 is the strain with the lowest bitterness, but when B, C, and D media are used, the yield is increased but the fruiting body obtained is bitter. .
[0010]
In the present invention, a highly bittering medium refers to a medium in which the bitterness of fruiting bodies obtained when artificially cultivating lyophilum ulmarium Lu 1-2 is + or higher by a sensory test, and the above B, C, D medium is there. The novel strain of the present invention refers to a strain in which the bitterness of fruiting bodies obtained by culturing the strain in a highly bittering medium is less than or equal to at least one kind of bittering medium.
[0011]
Next, breeding of the novel strain of the present invention will be described.
1. Selective breeding
Tissue isolation was performed on 112 fruit bodies of Riophyllum ulmarium occurring in nature to obtain 90 purely isolated mycelium strains. Next, a cultivation test was performed on the 90 strains using the B medium. Through the cultivation test, the fruiting bodies of 48 strains that formed fruiting bodies were subjected to a sensory test. Ten strains with excellent fruiting body shape, high yield and low bitterness were selected. Further, these 10 strains were subjected to cultivation tests in B, C, and D media, and lyophilum ulmarium Lu 1-13 was selected as a strain having the lowest bitterness suitable for industrial cultivation.
[0012]
Hereinafter, this lyophilum ulmarium will be described.
The lyophilum ulmarium Lu 1-13 strain was isolated by the present inventors from the fruiting bodies that had clustered in dead trees in Urabandai, Fukushima Prefecture. The characteristics of the fruiting bodies and spores are as follows. The fruiting body is crowded, the body is 5-1.5 cm in diameter, round or irregular, round mountain shape, the surface is smooth, wet, white to brownish cream color, often showing a slightly dark spot, old age Cracks may form in the center. The meat is white and grows in a wide variety of patterns. The handle is eccentric and curved, 3-7 × 1-2 cm, almost the same color as the umbrella, the top is white and fluffy or powdery. The spores were almost spherical, smooth, colorless, 4.5-5.5 × 3.5-4.5 μm, and the crest was white.
Comparing these characteristics with the description in Seiya Ito, "Japanese Fungi Journal," Vol. 5, No. 5 (1959, Yokendo Publishing), it is clear that the bacterium is lyophilum ulmarium.
[0013]
Next, the bacteriological properties of the lyophilum ulmarium Lu 1-13 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 7: Vigorous growth. The colony diameter is 40 mm. A large amount of white, dense hyphae and aerial hyphae. Day 10: Hyphae grow throughout the petri dish. Day 17: dense aerial hyphae form over the entire surface. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 7: Vigorous growth. The colony diameter is 36 mm. A large amount of white, dense hyphae and aerial hyphae. Day 10: Hyphae grow throughout the petri dish. Day 17: dense aerial hyphae form over the entire surface. The center of the colony is pale yellow, and others are white.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 7: small growth. The colony diameter is 26 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 17: Mycelia grow throughout the petri dish. Hyphae are dendritic, thin, white.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 7: Vigorous growth. The colony diameter is 42 mm. White, flocculent, dense hyphae, aerial hyphae. Day 10: Hyphae grow throughout the petri dish. A large amount of aerial mycelium is formed, and the mycelium is cottony and white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 7: Vigorous growth. The colony diameter is 36 mm. Hyphae are well-branched and stretched, and aerial hyphae are few. Day 10: Hyphae grow throughout the petri dish. A large amount of flocculent aerial hyphae. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 7: small growth. The colony diameter is 21 mm. The hyphae grow white and linear, forming radial colonies. Day 17: Mycelia grow throughout the petri dish. Produces a large amount of aerial hyphae. Mycelium is white.
(7) YpSs agar medium (cultured at 25 ° C)
Day 7: Vigorous growth. The colony diameter is 40 mm. A large amount of white, dense hyphae and aerial hyphae. Matte shape. Day 10: Hyphae grow throughout the petri dish. Produces a large amount of dense aerial hyphae. The mycelium turns white, but the medium turns yellow.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 7: small growth. Colony diameter is 18mm. The hyphae are white, grow short and matte, and have few aerial hyphae. The medium is browning, the browning radius is 40 mm. Day 17: Medium growth. The colony diameter is 37 mm. The browning radius is 42 mm. There is a marked difference in growth rate between new and old seeds.
(9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 7-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0014]
Next, lyophilum ulmarium Lu 1-13 strain and other lyophilum
Based on the fungal taxonomic fact that if the sex factors of both hyphae were different, the hyphae were different from each other, the difference in sex factors was examined by facing culture on an agar medium. .
The lyophilium ulmarium tested was lyophilium ulmarium
IFO 9637, liophilum ulmarium IFO 30525, liophilum ulmarium Lu 1-2, liophilum ulmarium Lu 1-8, liophilum ulmarium Lu 1-17, riophilum ulmarium M-8171, liophilum ulmarium ulmarium ulmarium. In addition, liophilum ulmarium SA is a isolate from commercially available lyophilum ulmarium fruit body.
Each of the above-mentioned dinuclear hypha of lyophilum ulmarium was cut out from the storage slant as a block of 3 × 3 × 3 mm, and each was inoculated in the center of the PGY agar medium in opposition to the dinuclear mycelium of lyophilum ulmarium Lu 1-13. After culturing at 25 ° C. (at 2 cm intervals) for 14 days, it was determined whether or not a band was formed between colonies of both strains. Table 2 shows the results. (+ When banding occurs,-when not occurring).
[0015]
[Table 2]

[0016]
As can be seen from Table 2, each of the strains was lyophilum ulmarium Lu.
All bands were generated in the confrontation culture with 1-13, from which it is clear that lyophilum ulmarium Lu 1-13 is a new strain.
As described above, as a new strain obtained by the selective breeding of the present invention, for example, lyophilum ulmarium Lu 1-13 can be mentioned, but the bitterness of the fruiting body obtained by cultivation in a highly bitter medium as in the case of the above strain is at least one. All of the strains exhibiting the characteristic that the bitterness is not more than almost no bitterness belong to the present invention.
[0017]
2. Breeding
The new strain of the present invention was obtained by cross breeding.
As described in JP-A-63-273467, lipophilum ulmarium M-8171 excellent in yield, growth rate and fruiting body morphology can be obtained by crossing lyophilum ulmarium Lu 1-8 and lyophilum ulmarium Lu 1-17. ing. In order to cross-breed strains having excellent cultivation characteristics and reduced bitterness of fruiting bodies, both strains of this lyophilum ulmarium Lu 1-8 and lyophilum ulmarium Lu 1-17 were used as parent strains.
Liophyllum ulmarium Lu 1-8 and lyophilum ulmarium
Lu 1-17 was grown on A medium, respectively, and fruiting bodies were generated by a usual operation. Spores were collected from the fruiting bodies, and germinated mononuclear hyphae were isolated. 57 strains were bred in total and 1653 strains were obtained. A cultivation test was performed on the 1653 strains using the B medium, and 80 strains having low bitterness were selected by sensory tests. Then, cultivation tests were performed on the B, C, and D media. About one kind of highly bittering medium, eight strains which are less than or equal to almost no bitterness are selected, and each of them is lyophilum ulmarium K-0429, lyophilum ulmarium K-0207, lyophilum ulmarium K-0202, lyophilum ulmarium K-0259, and liophilum ulmarium. K-1004, lyophilum ulmarium K-1257, lyophilum ulmarium K-6804, and liophilum ulmarium K-6806, respectively.
[0018]
Next, various bacteriological properties of the lyophilum ulmarium K-0429 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 5: Colony diameter 18 mm. A large amount of white, dense hyphae and aerial hyphae. Day 10: Colony diameter is 47 mm. Day 20: colony diameter 83 mm. It grows radially and produces dense aerial hyphae over the entire surface. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 40 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Colony diameter 55 mm. Day 20: Colony diameter is 66 mm. It produces dense aerial hyphae over the entire surface. A flocculent colony with white hyphae.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 28 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 20: colony diameter 52 mm. Hyphae are dendritic, thin, white.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 50 mm. White, flocculent, dense hyphae, aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It grows radially and produces aerial hyphae over the entire surface. Mycelium is white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 51 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow throughout the petri dish. A flocculent colony that produces large amounts of aerial hyphae. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 27 mm. The mycelium is white and forms dendritic colonies. Day 20: colony diameter 48 mm. This produces aerial hyphae. Dendritic colony with white hyphae.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 49 mm. White hyphae and aerial hyphae occur in flocculent colonies. Day 20: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 5: Small growth. Colony diameter is 18mm. The mycelium is white and produces large amounts of aerial hyphae. The medium turned brown with a browning radius of 34 mm. Day 15: Vigorous growth. Colony diameter is 64 mm. The entire surface of the medium is browned. There is a marked difference in growth rate between new and old seeds.
(9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 7-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0019]
Next, various bacteriological properties of the lyophilum ulmarium K-0207 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 61 mm. White, radial colonies, small amounts of aerial hyphae. Day 20: Mycelia grow throughout the petri dish. There are few aerial hyphae. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 53 mm. A large amount of white, dense hyphae and aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. Colonies are cottony and white.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 7: Vigorous growth. The colony diameter is 51 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 15: Colony diameter is 75 mm. Hyphae are dendritic, thin, white.
Day 20: Colony diameter is 77 mm.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Very active growth. The colony diameter is 58 mm. White, radial dense mycelium, aerial mycelium Day 15: Hyphae grow throughout the petri dish. Aerial mycelium forms, radially white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 66 mm. The colonies are flocculent and give rise to aerial hyphae. Day 15: Hyphae grow throughout the petri dish. Aerial hyphae occur in flocculent colonies. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 33 mm. The hyphae are white, slightly thin, elongate in dendrites, with aerial hyphae. Day 20: colony diameter 74 mm. Colonies are sparse and dendritic, mycelium white.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 65 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 63 mm. The mycelium is white and produces large amounts of aerial hyphae. The medium is browning and the browning radius is 65 mm. Day 15: Mycelia grow throughout the petri dish. A large amount of aerial mycelium is produced, and the entire surface of the medium turns brown. There is a marked difference in growth rate between new and old seeds.
(9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 7-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0020]
Next, various bacteriological properties of the lyophilum ulmarium K-0202 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 60 mm. White, radial colonies, small amounts of aerial hyphae. Day 20: Mycelia grow throughout the petri dish. There are few aerial hyphae. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 59 mm. A large amount of white, dense hyphae and aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. Colonies are cottony and white.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 5: Vigorous growth. Colony diameter is 16mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 15: Colony diameter is 62 mm. Hyphae are dendritic, thin, white.
Day 20: Colony diameter is 77 mm.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Very active growth. The colony diameter is 66 mm. White, radial dense mycelium, aerial mycelium Day 15: Mycelia grow throughout the petri dish. Aerial mycelium forms, radially white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 68 mm. The colonies are flocculent and give rise to aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Aerial hyphae occur in flocculent colonies. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 33 mm. The hyphae are white, slightly thin, elongate in dendrites, with aerial hyphae. Day 20: colony diameter is 62 mm. Colonies are sparse and dendritic, mycelium white.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 68 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 61 mm. The mycelium is white and produces large amounts of aerial hyphae. Medium browned. Day 15: Mycelia grow throughout the petri dish. A large amount of aerial mycelium is produced, and the entire surface of the medium turns brown. There is a marked difference in growth rate between new and old seeds. (9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 7 to 8.5. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0021]
Next, various bacteriological properties of the lyophilum ulmarium K-0259 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 5: Colony diameter 24 mm. A large number of white, dense hyphae and aerial hyphae. Day 10: colony diameter 64 mm. Day 15: Mycelia grow throughout the petri dish. It grows radially and produces dense aerial hyphae over the entire surface. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 61 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Colony diameter is 85 mm. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. A flocculent colony with white hyphae.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 48 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 20: Colony diameter is 75 mm. Hyphae are dendritic, thin, white.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 65 mm. White, flocculent, dense hyphae, aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It grows radially and produces aerial hyphae over the entire surface. Mycelium is white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 67 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow throughout the petri dish. A flocculent colony that produces large amounts of aerial hyphae. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 39 mm. The mycelium is white and forms dendritic colonies. Day 20: Colony diameter is 66 mm. This produces aerial hyphae. Dendritic colony with white hyphae.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 63 mm. White hyphae and aerial hyphae occur in flocculent colonies. Day 20: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 5: Small growth. The colony diameter is 29 mm. The mycelium is white and produces large amounts of aerial hyphae. Medium browned. Day 15: Vigorous growth. The colony diameter is 88 mm. The entire surface of the medium is browned. There is a marked difference in growth rate between new and old seeds.
(9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0022]
Next, various bacteriological properties of the lyophilum ulmarium K-1004 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. Colony diameter is 64 mm. White, radial colonies, small amounts of aerial hyphae. Day 20: Mycelia grow throughout the petri dish. There are few aerial hyphae. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. Colony diameter is 64 mm. A large amount of white, dense hyphae and aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. Colonies are cottony and white.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 5: Vigorous growth. The colony diameter is 14 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 15: Colony diameter is 60 mm. Hyphae are dendritic, thin, white. Day 20: Colony diameter is 75 mm.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Very active growth. The colony diameter is 83 mm. White, radial dense mycelium, aerial mycelium Day 15: Mycelia grow throughout the petri dish. Aerial mycelium forms, radially white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 81 mm. Colonies are flocculent and give rise to abundant aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Aerial hyphae occur in flocculent colonies. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 23 mm. The mycelium is white, slightly thin, dendritic, accompanied by aerial mycelia. Day 20: colony diameter 45 mm. Colonies are sparse and dendritic. Mycelium is white.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 80 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 68 mm. The mycelium is white and produces large amounts of aerial hyphae. Medium browned. Day 15: Mycelia grow throughout the petri dish. A large amount of aerial mycelium is produced, and the entire surface of the medium turns brown. There is a marked difference in growth rate between new and old seeds. (9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-7. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0023]
Next, various bacteriological properties of the lyophilum ulmarium K-1257 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 71 mm. White, radial colonies, small amounts of aerial hyphae. Day 20: Mycelia grow throughout the petri dish. There are few aerial hyphae. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 66 mm. A large amount of white, dense hyphae and aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. Colonies are cottony and white.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 5: Vigorous growth. The colony diameter is 26 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 15: Colony diameter 71 mm. Hyphae are dendritic, thin, white. Day 20: colony diameter 81 mm.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Very active growth. The colony diameter is 79 mm. White, radial dense mycelium, aerial mycelium Day 15: Mycelia grow throughout the petri dish. Aerial mycelium forms, radially white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. The colony diameter is 79 mm. Colonies are flocculent and give rise to abundant aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Aerial hyphae occur in flocculent colonies. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 37 mm. The hyphae are white, slightly thin, elongate in dendrites, with aerial hyphae. Day 20: Colony diameter is 68 mm. Colonies are sparse and dendritic. Mycelium is white.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Extremely vigorous growth. Colony diameter is 64 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 70 mm. The mycelium is white and produces large amounts of aerial hyphae. Medium browned. Day 15: Mycelia grow throughout the petri dish. A large amount of aerial mycelium is produced, and the entire surface of the medium turns brown. There is a marked difference in growth rate between new and old seeds. (9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0024]
Next, the mycological properties of the lyophilum ulmarium K-6804 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 5: Colony diameter 25 mm. A large amount of white, dense hyphae and aerial hyphae. Day 10: colony diameter 61 mm. Day 15: Colony diameter 83 mm. It grows radially and produces dense aerial hyphae over the entire surface. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 61 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Colony diameter 86 mm. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. A flocculent colony with white hyphae.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 49 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 20: colony diameter is 76 mm. Hyphae are dendritic, thin, white.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 70 mm. White, flocculent, dense hyphae, aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It grows radially and produces aerial hyphae over the entire surface. Mycelium is white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 74 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow throughout the petri dish. A flocculent colony that produces large amounts of aerial hyphae. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 29 mm. The mycelium is white and forms dendritic colonies. Day 20: colony diameter 53 mm. This produces aerial hyphae. Dendritic colony with white hyphae.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 72 mm. White hyphae and aerial hyphae occur in flocculent colonies. Day 20: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 5: Small growth. The colony diameter is 28 mm. The mycelium is white and produces large amounts of aerial hyphae. Medium browned. Day 15: Mycelia grow throughout the petri dish. The entire surface of the medium is browned. There is a marked difference in growth rate between new and old seeds.
(9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0025]
Next, the bacteriological properties of the lyophilum ulmarium K-6806 strain will be described.
(1) Malt extract agar medium (cultured at 25 ° C)
Day 5: Colony diameter is 19 mm. Produces a large amount of white, dense hyphae and aerial hyphae. Day 10: colony diameter 59 mm. Day 15: Colony diameter is 81 mm. It grows radially and produces dense aerial hyphae over the entire surface. Mycelium is white.
(2) Potato-glucose agar medium (cultured at 25 ° C)
Day 10: Medium growth. The colony diameter is 55 mm. A large amount of white, dense hyphae and aerial hyphae. Day 15: Colony diameter is 80 mm. Day 20: Mycelia grow throughout the petri dish. It produces dense aerial hyphae over the entire surface. A flocculent colony with white hyphae.
(3) Tzapek Dox agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 38 mm. Very few mycelia and aerial mycelia elongate like dendrites. Day 20: Colony diameter is 66 mm. Hyphae are dendritic, thin, white.
(4) Sabouraud agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 66 mm. White, flocculent, dense hyphae, aerial hyphae. Day 20: Mycelia grow throughout the petri dish. It grows radially and produces aerial hyphae over the entire surface. Mycelium is white.
(5) Oatmeal agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 71 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow throughout the petri dish. A flocculent colony that produces large amounts of aerial hyphae. Mycelium is white.
(6) Synthetic mucor agar medium (cultured at 25 ° C)
Day 10: Small growth. The colony diameter is 31 mm. The mycelium is white and forms dendritic colonies. Day 20: colony diameter 61 mm. This produces aerial hyphae. Dendritic colony with white hyphae.
(7) YpSs agar medium (cultured at 25 ° C)
Day 10: Vigorous growth. The colony diameter is 70 mm. White hyphae and aerial hyphae occur in flocculent colonies. Day 20: Mycelia grow throughout the petri dish. Produces a large amount of dense aerial hyphae. Mycelium is white.
(8) Phenoloxidase assay medium (cultured at 25 ° C)
Day 5: Small growth. The colony diameter is 28 mm. The mycelium is white and produces large amounts of aerial hyphae. Medium browned. Day 15: Vigorous growth. The colony diameter is 86 mm. The entire surface of the medium is browned. There is a marked difference in growth rate between new and old seeds.
(9) Optimal growth temperature
A disc-shaped inoculum having a diameter of 5 mm was inoculated on a PGY agar medium and cultured at each temperature for 12 days, and then the colony diameter was measured. As a result, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 ° C. and 35 ° C.
(10) Optimal growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, sterilize, adjust to each pH with acid or alkali, inoculate the inoculum, and incubate at 25 ° C for 15 days. After that, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Moreover, the pH range in which growth was possible was pH 3.5 to 10.
[0026]
Next, lyophilum ulmarium K-0429 strain, lyophilum ulmarium K-0207 strain, lyophilum ulmarium K-0202 strain, lyophilum ulmarium K-0259 strain, lyophilum ulmarium K-1004 strain, lyophilum ulmarium K-1257 strain, and lyophilum ulmarium K-68 strain. Strain and the lipophilum ulmarium K-6806 strain and the other lipophilum ulmarium strain were determined to be different from each other based on the fungal taxonomic fact that if the two hyphae had different sex factors, the hyphae were different hyphae. Was examined by confronting culture on an agar medium.
The lyophilium ulmarium tested was lyophilium ulmarium
IFO 9637, liophilum ulmarium IFO 30525, riophilum ulmarium Lu 1-2, liophilum ulmarium Lu 1-8, liophilum ulmarium Lu 1-17, riophilum ulmarium M-8171, liophilum 13-ulmarium ulmarium ullumium It is.
[0027]
Each of the above-mentioned dinuclear mycelia of lyophilum ulmarium was cut out as a 3 × 3 × 3 mm block from the preservation slant, and each was placed on the center of a PGY agar medium, and lyophilum ulmarium K-0429 strain, lyophilum ulmarium,
K-0207 strain, lyophilum ulmarium K-0202 strain, lyophilum ulmarium K-0259 strain, lyophilum ulmarium K-1004 strain, lyophilum ulmarium K-1257 strain, lyophilum ulmarium K-6804 strain, or lyophilum ulmarium K-6806 strain After inoculation (at 2 cm intervals) at 25 ° C. for 14 days, it was determined whether or not a band line was formed between colonies of both strains. The results are shown in Tables 3 and 4. (+ When banding occurs,-when not occurring).
[0028]
[Table 3]

[0029]
[Table 4]

[0030]
As can be seen from Tables 3 and 4, Liophyllum ulmarium K-0429, Liophyllum ulmarium K-0207, Liophyllum ulmarium K-0202, Liophyllum ulmarium K-0259, Liophyllum ulmarium K-1004, Liophyllum ulmarium K-1257, Liophyllum ulmarium K-68. , And Liophyllum ulmarium K-6806 all produce banding in facing culture with other strains, which clearly shows that both strains are new strains.
[0031]
As described above, new strains bred by the cross of the present invention include, for example, lyophilum ulmarium K-0429, lyophilum ulmarium K-0207, lyophilum ulmarium K-0202, lyophilum ulmarium K-0259, lyophilum ulmarium K-1004, lyophilum Ulmarium K-1257, lyophilum ulmarium K-6804, lyophilum ulmarium K-6806, and the like, but the bitterness of the fruiting bodies obtained by cultivation in a highly bitter medium in the same manner as the above strains has at least one type of high bitterness. All strains of the medium exhibiting a characteristic of not more than almost no bitterness belong to the present invention.
[0032]
The new strain according to the present invention has the property that the bitterness of the fruit body is at least one kind of highly bitter medium even when cultivated in a highly bitter medium during artificial cultivation as described above, and has a property that the bitter taste is hardly felt. It has a stable and low bitterness of the fruiting body regardless of the type of medium. Tables 5 and 6 show the bitterness and yield of the fruit body at the optimal harvest time obtained by artificially cultivating the new strains of the present invention using the A, B, C, and D media. Table 6 also shows the results of lyophilum ulmarium SA described above.
[0033]
[Table 5]

[0034]
[Table 6]

[0035]
As shown in Tables 5 and 6, lyophilum ulmarium Lu1-13, lyophilum ulmarium K-0429, lyophilum ulmarium K-0207, lyophilum ulmarium K-0202, lyophilum ulmarium K-0259, lyophilum ulmarium K-1004, lyophilum ulmarium K- 1257, Liophyllum ulmarium K-6804, and Liophyllum ulmarium K-6806 are strains of conventional artificially cultivable strains, which show no bitterness even when a medium in which the fruiting body at the appropriate time for harvesting exhibits bitterness shows no bitterness, and the fruiting body yield , But also increases the bitterness, so that a bittering-free fruiting body can be stably obtained at a high yield even if a highly bittering medium which has been conventionally difficult to use is used.
[0036]
Moreover, the mycelium produced by inoculating the above strain into a medium does not exhibit bitterness, and is suitable for use in foods. In addition, liophilum ulmarium mycelium has a large amount of dietary fiber and is also known to have an anticancer effect, and foods using the mycelium are particularly useful for maintaining health.
[0037]
The new strains bred in the present invention, lyophilum ulmarium Lu 1-13, lyophilum ulmarium K-0429, lyophilum ulmarium K-0207, lyophilum ulmarium K-0202, liophilum ulmarium K-0259, liophilum ulmarium K-1004, liophilum 1257-ulmarium K- , Lyophyllum ulmarium K-6804 and Lyophyllum ulmarium K-1806 are respectively Lyophyllum ulmarium Lu 1-13, Lyophyllum ulmarium K-0429, Lyophyllum ulmarium K-0207, Lyophyllum ulmarium K-0202, Lyophyllum ulmarium Kullum ulmarium Kylum-4 ulmarium Kulum ulmarium Kulum ulmarium Kulum ulmarium Kulum 4 , Lyophyllum ulmarium K-1257, Lyophyllum ulmarium K-6804, and Lyophyllum ulmarium K-6806. No. 12571 (FERMP-12571), No. 12570 (FERM P-12570), No. 12569 (FERM)
P-12569), Microtechnical Laboratories No. 12980 (FERM P-12980), Microtechnical Research Microorganisms No. 12981 (FERM P-12981), Microtechnical Research Microorganisms No. 12982 (FERM P-12982), No. 12983 (Ferm)
P-12983), No. 12984 (FERM P-12984) and No. 12985 (FERM P-12985).
[0038]
The bitter component in the lyophilum ulmarium fruit body can also be quantified as follows.
The edible parts (casa and stalks) in the lyophilum ulmarium fruit body are freeze-dried and crushed to obtain a freeze-dried powder of the fruit body. 0.5 g (dry weight) of the freeze-dried powder was extracted by shaking with 35 ml of ethyl acetate at 25 ° C. for 24 hours, filtered, concentrated under reduced pressure, dissolved in 1 ml of methanol while heating, and used as an extracted sample.
From the above 200 μl sample, a bitter component fraction is fractionated as follows using FPLC (Pharmacia Fine Chemicals). The column used was a solvent of 85% methanol (Kanto Chemical; liquid chromatography), 1.0 ml / min, room temperature, detection wavelength of 210 nm using Lober RP-8 (manufactured by Merck, φ1.0 × 24 cm). Under the conditions, a fraction with an elution time of 10 to 45 minutes is collected.
The fractionated fraction was again concentrated under reduced pressure, dissolved in 1.0 ml of a solvent of methanol: ethanol: water = 3: 4: 4, and 50 μl thereof was bitter by HPLC [LC-6A system manufactured by Shimadzu Corporation]. Separate the component fractions. Column is μ Bondapak C ₁₈ Methanol: ethanol: water = 3: 4: 4 solvent (Kanto Chemical; methanol for liquid chromatography, Nacalai Tesque; special grade ethanol) using [Waters, φ0.39 × 30 cm]; The dry weight of the fraction with an elution time of 32.5 to 44.0 minutes is measured under the conditions of 7 ml / min, a temperature of 40 ° C. and a detection wavelength of UV 210 nm. The component eluted in this fraction (hereinafter abbreviated as fraction A) has a strong bitter taste in the sensory test, and the bitter taste of the lyophilum ulmarium fruit body is due to this fraction A.
[0039]
The present inventors have found the cottonseed husk used in the medium of the present invention as a base material useful for artificial cultivation of mushrooms, which can be obtained at low cost and significantly increases the yield of mushrooms. When this cottonseed shell is used for a culture medium, it may be used by mixing it with another medium base material, or may be used alone. The use of cottonseed husks is most effective when used at 15 to 60% of the dry weight of the medium, but the amount used is not limited thereto. Mushrooms that can be cultivated with high yield by using this cottonseed shell as a medium include, for example, lyophilum ulmarium of the present invention, shiitake, oyster mushroom, enokitake, maitake, and the like.For example, using these mushroom strains, You only have to do artificial cultivation.
[0040]
【Example】
Examples of artificial cultivation of a new strain of lyophilum ulmarium according to the present invention will be described below, but the present invention is not limited only to the scope of the following examples.
Examples 1 to 3 below are examples given as reference examples of the present invention.
[0041]
Example 1
100 ml of liquid PGY medium (PGY agar medium without agar) was inoculated with lyophilum ulmarium Lu 1-13 (FERM P-12571) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 35 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 40 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 168 g, and no bitterness was felt in the sensory test.
[0042]
In addition, the sensory test of the fruiting body was performed as follows. The base of the handle of the fruiting body obtained in the above bottle cultivation was removed, divided into single pieces, washed lightly with water, and drained well. On the other hand, add 5 ml of salad oil to a frying pan, spread the whole, heat the oil sufficiently over medium heat, add 100 g of fruit body (wet weight), stir with chopsticks and cook for 2 minutes or more. Stir fry until dry. The oil-fried fruit body was cooled to room temperature, and bitterness was determined by 10 panelists using quinine sulfate as a standard substance.
[0043]
Example 2
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium Lu 1-13.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 35 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain fruit body primordia, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain fruit body.
The yield of the obtained fruit body was 143 g, and no bitterness was felt in the sensory test of the fruit body.
[0044]
Example 3
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium Lu 1-13.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 37 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 157 g, and no bitterness was felt in the sensory test of the fruit body.
[0045]
Example 4
Liophyllum ulmarium K-0429 (FERM P-12570) was inoculated into 100 ml of liquid PGY medium and cultured at 25 ° C for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 32 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 43 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain fruit body primordia, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain fruit body.
The yield of the obtained fruit body was 184 g, and no bitterness was felt in the sensory test of the fruit body.
[0046]
Example 5
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of Lyophilum ulmarium K-0429.
When the culture medium was cultured for 28 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 37 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain fruit body primordia, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain fruit body.
The yield of the obtained fruit body was 157 g, and no bitterness was felt in the sensory test of the fruit body.
[0047]
Example 6
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0429.
When the culture medium was cultured for 28 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 37 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 164 g, and no bitterness was felt in the sensory test of the fruit body.
[0048]
Example 7
100 ml of liquid PGY medium was inoculated with lyophilum ulmarium K-0207 (FERM P-12569) and cultured at 25 ° C for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 40 days in a dark place at 25 ° C and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 35 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 10 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 14 days to obtain a fruit body.
The yield of the obtained fruit body was 188 g, and bitterness was not felt in the sensory test of the fruit body.
[0049]
Example 8
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0207.
When the culture medium was cultured for 34 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 31 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 14 days to obtain a fruit body.
The yield of the obtained fruiting body was 161 g, and bitterness was not felt in the sensory test of the fruiting body.
[0050]
Example 9
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0207.
When the culture medium was cultured for 33 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 32 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 11 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 178 g, and bitterness was not felt in the sensory test of the fruit body.
[0051]
Example 10
Liophyllum ulmarium K-0202 (FERM P-12980) was inoculated into 100 ml of liquid PGY medium and cultured at 25 ° C for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 45 days in the dark at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 35 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, excess tap water is discarded, and the conditions are maintained at 15 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 8 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 187 g, and no bitterness was felt in the sensory test of the fruit body.
[0052]
Example 11
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0202.
When the culture medium was cultured for 40 days in a dark place at 25 ° C and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 40 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, excess tap water is discarded, and the conditions are maintained at 15 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 8 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 12 days to obtain a fruit body.
The yield of the obtained fruit body was 154 g, and bitterness was not felt in the sensory test of the fruit body.
[0053]
Example 12
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0202.
When the culture medium was cultured for 36 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 44 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, excess tap water is discarded, and the conditions are maintained at 15 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 8 days. The cultivation was continued to obtain fruit body primordia, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain fruit body.
The yield of the obtained fruit body was 162 g, and bitterness was not felt in the sensory test of the fruit body.
[0054]
Example 13
Liophyllum ulmarium K-0259 (FERM P-12981) was inoculated into 100 ml of liquid PGY medium and cultured at 25 ° C for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 45 days in the dark at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 35 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, excess tap water is discarded, and the conditions are maintained at 15 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 8 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 12 days to obtain a fruit body.
The yield of the obtained fruit body was 192 g, and no bitterness was felt in the sensory test of the fruit body.
[0055]
Example 14
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0259.
When the culture medium was cultured for 35 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 45 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° -95% humidity and 20 lux illuminance for 8 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 162 g, and bitterness was not felt in the sensory test of the fruit body.
[0056]
Example 15
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water is pressed into a 850 ml wide-mouth bottle made of polypropylene, and a hole with a diameter of 1 cm is directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-0259.
When the culture medium was cultured for 37 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 43 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, excess tap water is discarded, and the conditions are maintained at 15 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 8 days. The cultivation was continued to obtain fruit body primordia, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain fruit body.
The yield of the obtained fruit body was 165 g, and no bitterness was felt in the sensory test of the fruit body.
[0057]
Example 16
Liophyllum ulmarium K-1004 (FERM P-12982) was inoculated into 100 ml of liquid PGY medium and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 50 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 11 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 15 days to obtain a fruit body.
The yield of the obtained fruit body was 188 g, and bitterness was not felt in the sensory test of the fruit body.
[0058]
Example 17
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole 1 cm in diameter downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-1004.
When the culture medium was cultured for 32 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 48 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 10 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 15 days to obtain a fruit body.
The yield of the obtained fruit body was 158 g, and no bitterness was felt in the sensory test of the fruit body.
[0059]
Example 18
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-1004.
When the culture medium was cultured for 28 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 52 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 11 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 16 days to obtain a fruit body.
The yield of the obtained fruiting body was 161 g, and bitterness was not felt in the sensory test of the fruiting body.
[0060]
Example 19
Liophyllum ulmarium K-1257 (FERM P-12983) was inoculated into 100 ml of liquid PGY medium, and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 50 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 11 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 15 days to obtain a fruit body.
The yield of the obtained fruit body was 186 g, and bitterness was not felt in the sensory test of the fruit body.
[0061]
Example 20
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-1257.
When the culture medium was cultured for 32 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 48 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 16 days to obtain a fruit body.
The yield of the obtained fruit body was 160 g, and no bitterness was felt in the sensory test of the fruit body.
[0062]
Example 21
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-1257.
When the culture medium was cultured for 28 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 52 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 10 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 14 days to obtain a fruit body.
The yield of the obtained fruit body was 157 g, and no bitterness was felt in the sensory test of the fruit body.
[0063]
Example 22
100 ml of liquid PGY medium was inoculated with lyophilum ulmarium K-6804 (FERM P-12984) and cultured at 25 ° C for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 30 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 10 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 16 days to obtain a fruit body.
The yield of the obtained fruit body was 195 g, and no bitterness was felt in the sensory test of the fruit body.
[0064]
Example 23
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-6804.
When the culture medium was cultured for 28 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 32 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 10 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 15 days to obtain a fruit body.
The yield of the obtained fruit body was 163 g, and no bitterness was felt in the sensory test of the fruit body.
[0065]
Example 24
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-6804.
When the culture medium was cultured for 28 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 32 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 16 days to obtain a fruit body.
The yield of the obtained fruit body was 171 g, and bitterness was not felt in the sensory test of the fruit body.
[0066]
Example 25
Liophyllum ulmarium K-6806 (FERM P-12985) was inoculated into 100 ml of liquid PGY medium and cultured at 25 ° C for 10 days to obtain a liquid inoculum. On the other hand, a mixture of 130 g of corn cob meal, 60 g of Mamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water was pressed into a 850 ml wide-mouth bottle made of polypropylene, and pressed downward from the center of the mouth of the bottle. After drilling a hole 1 cm in diameter, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum.
When the culture medium was cultured for 32 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 38 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 15 days to obtain a fruit body.
The yield of the obtained fruit body was 191 g, and bitterness was not felt in the sensory test of the fruit body.
[0067]
Example 26
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole with a diameter of 1 cm downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-6806.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Furthermore, culturing was continued for 40 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 15 days to obtain a fruit body.
The yield of the obtained fruit body was 162 g, and bitterness was not felt in the sensory test of the fruit body.
[0068]
Example 27
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium K-6806.
When the culture medium was cultured for 31 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 39 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 14 days to obtain a fruit body.
The yield of the obtained fruit body was 165 g, and no bitterness was felt in the sensory test of the fruit body.
[0069]
Comparative Example 1
A mixture of 130 g of corn cob meal, 60 g of Makamekawa and 30 g of bran mixed together and adjusted to a water content of 62% with tap water is pressed into a 850 ml polypropylene wide-mouth bottle, and the diameter is directed downward from the center of the mouth of the bottle. After drilling a 1 cm hole, the culture medium was autoclaved at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium Lu 1-2.
When the culture medium was cultured for 35 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 50 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The cultivation was continued to obtain fruit body primordia, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain fruit body.
The yield of the obtained fruit body was 178 g, and bitterness was felt in a sensory test of the fruit body.
[0070]
Control Example 2
50 g of softwood sawdust, 50 g of crushed cottonseed shell, and 100 g of rice bran were mixed well and adjusted to a water content of 63% with tap water, and pressed into a 850 ml wide-mouth bottle made of polypropylene. After drilling a hole 1 cm in diameter downward, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium Lu 1-2.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 55 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply the water, the excess tap water was discarded, and the conditions were maintained at 15 ° C., 90 to 95% humidity, and 20 lux illuminance for 9 days. The culture was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 14 days to obtain a fruit body.
The yield of the obtained fruit body was 140 g, and bitterness was felt in a sensory test of the fruit body.
[0071]
Control Example 3
A mixture of 100 g of coniferous sawdust and 70 g of Mekamawa mixed well and adjusted to a water content of 63% with tap water was pressed into a 850 ml polypropylene wide-mouth bottle, and a hole with a diameter of 1 cm was directed downward from the center of the mouth of the bottle. After opening the culture medium, the culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of lyophilum ulmarium Lu 1-2.
When the culture medium was cultured for 30 days in a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia spread throughout the bottle. Further, culturing was continued for 55 days under the same conditions to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body and adding 20 ml of tap water to sufficiently supply water, the excess tap water was discarded, and 15 days at 90 ° C., humidity of 90 to 95%, and illuminance of 20 lux for 10 days. The cultivation was continued to obtain the fruit body primordium, and the illuminance was further increased to 200 lux, and the culture was continued for 13 days to obtain a fruit body.
The yield of the obtained fruit body was 153 g, and the sensory test of the fruit body showed bitterness.
[0072]
【The invention's effect】
As described above, according to the present invention, it is possible to obtain a lyophilum ulmarium fruit body with reduced bitterness of fruit bodies even when cultivated using a highly bittering medium that could not be used conventionally, and Thus, it is possible to provide a high-yield and high-quality lyophilum ulmarium fruit body and mycelium.

Claims (1)

The bitterness of fruiting bodies obtained by cultivation in a highly bittering medium is less bitter than at least one kind of high bittering medium than 0.000008M quinine sulfate, and is selected from the following strains or mutants thereof. A new lyophilum ulmarium strain, which is a new ulmarium strain.
Lyophilum ulmarium K-0429 (FERM P-12570),
Lyophilum ulmarium K-0207 (FERM P-12569),
Lyophilum ulmarium K-0202 (FERM P-12980),
Lyophilum ulmarium K-0259 (FERM P-12981),
Lyophilum ulmarium K-1004 (FERM P-12982),
Lyophilum ulmarium K-1257 (FERM P-12983),
Lyophilum ulmarium K-6804 (FERM P-12984),
Liophyllum ulmarium K-6806 (FERM P-12985)