JP3436768B2 - Culture and cultivation method of new strain - Google Patents
Culture and cultivation method of new strainInfo
- Publication number
- JP3436768B2 JP3436768B2 JP23163492A JP23163492A JP3436768B2 JP 3436768 B2 JP3436768 B2 JP 3436768B2 JP 23163492 A JP23163492 A JP 23163492A JP 23163492 A JP23163492 A JP 23163492A JP 3436768 B2 JP3436768 B2 JP 3436768B2
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- Prior art keywords
- medium
- culture
- ulmarium
- day
- hyphae
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【0001】[0001]
【産業上の利用分野】本発明は、担子菌の新菌株に関
し、更に詳しくはリオフィラム ウルマリウム( Lyoph
yllum ulmarium )の新菌株の培養方法及び子実体の栽培
方法に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a new strain of basidiomycete, more specifically, Lyophyllum ulmarium
and a method for cultivating fruiting bodies of a new strain of yllum ulmarium).
【0002】[0002]
【従来の技術】リオフィラム ウルマリウムは、自然界
においては秋期に種々の広葉樹の枯れ木に叢生あるいは
離生しており、従来より形や歯切れの良い肉質のため、
極めて美味なきのことして採食されてきた。また、近年
ではオガクズに米糠やその他の栄養源を配合した培養基
を用いて、ビン又は箱で栽培を行う菌床人工栽培法が確
立され、季節に関係なく一年を通じて安定してきのこを
収穫できるようになってきた。ここでリオフィラム ウ
ルマリウムの菌床人工栽培を簡略に説明すれば、ビン又
は箱に培養基を詰め蒸気滅菌したものに、予め同様の培
養基に菌糸を繁殖させておいた種菌を接種し、所定の日
数培養した後発生操作を行い、子実体を得て収穫して1
サイクルが終了する。栽培に使用する菌株として数種の
菌株が市販あるいは技術提携により供与されているが、
使用する菌株により様々な形態や栽培特性がある(特開
昭63−273467号)。2. Description of the Related Art Riofilum ulmarium naturally grows into dead trees of various broad-leaved trees in the autumn, or vegetatively grows. Because of its shape and crispy flesh quality,
It has been eaten as an extremely delicious mushroom. In addition, in recent years, using a culture medium containing rice bran and other nutrients in sawdust, a fungal bed artificial cultivation method has been established in which cultivation is performed in bottles or boxes, and mushrooms can be stably harvested throughout the year regardless of the season. It's starting to happen. Briefly explaining artificial bed cultivation of riophyllum ulmarium here, a bottle or box filled with culture medium and steam sterilized is inoculated with inoculum in which hyphae have been propagated in the same culture medium beforehand, and cultured for a predetermined number of days. After that, carry out the generation operation to obtain fruit bodies and harvest 1
The cycle ends. Several strains are commercially available or provided by technical cooperation as strains used for cultivation,
There are various forms and cultivation characteristics depending on the strain used (Japanese Patent Laid-Open No. 63-273467).
【0003】[0003]
【発明が解決しようとする課題】しかしながら、現在使
用されているリオフィラム ウルマリウム菌株のすべて
は収穫時の子実体中に苦味成分を含有しており、子実体
収量を増加させる高苦味化培地では食用に適さないほど
の苦味をもつ。また、この苦味成分は培地の種類により
変動を示し、苦味の強さに大きく差が生じる。一般にき
のこは素材の味を重視する蔬菜的使用方法が主流なた
め、苦味成分が多く含有されることは著しく商品価値を
低下させる結果となる。このために、たとえ子実体の収
穫量を著しく増加させる培地基材であっても、同時に苦
味成分を増加させる培地、すなわち高苦味化培地は使用
できないのが現状である。本発明の目的は、上記現状に
かんがみ、これら高苦味化培地で栽培しても収穫時の子
実体中の苦味成分が食用に供した場合苦味をほとんど感
じない程度以下の新菌株の培養及び栽培方法を提供する
ことにある。However, all of the currently used Riophyllum ulmarium strains contain bitterness components in the fruiting bodies at the time of harvesting, and are edible in a high bittering medium that increases the yield of fruiting bodies. Has an unsuitable bitterness. In addition, the bitterness component varies depending on the type of medium, and there is a large difference in the bitterness intensity. Generally, mushrooms are mainly used in a vegetable-like manner, which places importance on the taste of the raw material, so that a large amount of bitterness components results in a marked decrease in commercial value. For this reason, the present situation is that even a medium base material that significantly increases the yield of fruiting bodies, a medium that simultaneously increases the bitterness component, that is, a high bitterness medium cannot be used. In view of the present situation, the object of the present invention is to cultivate and culture these new bitterness strains with little bitterness when the bitterness component in the fruiting body at harvest is edible even when cultivated in these high bittering medium. To provide a method.
【0004】[0004]
【課題を解決するための手段】本発明を概説すれば、本
発明の第1の発明はリオフィラム ウルマリウム新菌株
の培養方法に関する発明であって、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、0.000008M 硫酸キニーネよ
りも苦くなく、かつ下記の菌株、又はこれらの変異株か
ら選択されるリオフィラム ウルマリウム新菌株を培地
に接種し、菌糸体を生成させることを特徴とする。
リオフィラム ウルマリウム K−0429(FERM
P−12570)、
リオフィラム ウルマリウム K−0207(FERM
P−12569)、
リオフィラム ウルマリウム K−0202(FERM
P−12980)、
リオフィラム ウルマリウム K−0259(FERM
P−12981)、
リオフィラム ウルマリウム K−1004(FERM
P−12982)、
リオフィラム ウルマリウム K−1257(FERM
P−12983)、
リオフィラム ウルマリウム K−6804(FERM
P−12984)、
リオフィラム ウルマリウム K−6806(FERM
P−12985)
また本発明の第2の発明はリオフィラム ウルマリウム
新菌株の子実体の栽培方法に関する発明であって、上記
第1の発明におけるリオフィラム ウルマリウム新菌株
を培地に接種し、子実体を生成させることを特徴とす
る。SUMMARY OF THE INVENTION The present invention will be summarized. The first invention of the present invention is a new strain of riophyllum ulmarium.
The invention relating to the culturing method, wherein the bitterness of the fruiting body obtained by culturing in the high bittering medium is less bitter than 0.000008M quinine sulfate for at least one high bittering medium, and the strains described below, or medium Lyophyllum ulmarium new strains are selected from these mutant strains
Inoculated into, characterized by Rukoto to produce a mycelium. Riofilum Ulmarium K-0429 (FERM
P-12570), Riofilum ulmarium K-0207 (FERM
P-12569), Riofilum ulmarium K-0202 (FERM
P-12980), Riofilum ulmarium K-0259 (FERM
P-12981), Riofilum ulmarium K-1004 (FERM
P-12982), Riofilum ulmarium K-1257 (FERM
P-12983), Riofilum ulmarium K-6804 (FERM
P-12984), Riofilum ulmarium K-6806 (FERM
A second invention of the P-12985) The present invention is an invention relating to a method of cultivating a fruit body of Lyophyllum ulmarium novel strain was inoculated to the medium Lyophyllum ulmarium novel strain in the first invention, the fruiting bodies generate It is characterized by
【0005】本発明者らはリオフィラム ウルマリウム
子実体の官能検査を行い、子実体の苦味と栽培培地との
関係を明らかにし、子実体の苦味を増加させる培地、す
なわち高苦味化培地を用い人工栽培を行っても、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リオフィ
ラム ウルマリウム新菌株を育種することに成功し、本
発明を完成させた。The present inventors conducted a sensory test on the fruiting body of Riofilum ulmarium to clarify the relationship between the bitterness of the fruiting body and the cultivation medium, and artificially cultivate it using a medium for increasing the bitterness of the fruiting body, that is, a high bittering medium. The present invention succeeded in breeding a new strain of Riofilum ulmarium, in which the bitterness of the fruiting body thus obtained was at least as high as the bitterness in at least one type of high-bittering medium, and the present invention was completed.
【0006】以下、本発明を詳細に説明する。リオフィ
ラム ウルマリウム菌株として、特開昭63−2734
67号公報に記載のリオフィラム ウルマリウム Lu
1−2(FERM P−12584)、リオフィラム
ウルマリウム Lu 1−8(FERM BP−14
16)、リオフィラム ウルマリウム Lu 1−17
(FERM BP−1417)、リオフィラム ウルマ
リウム M−8171(FERM BP−1415)を
使用し、該公報に記載の方法により、各菌株の人工栽培
を行い、収穫適期の子実体を収穫し、子実体の官能検査
を行った。人工栽培の培地としては、該公報に記載の針
葉樹オガクズ50g、広葉樹オガクズ50g、米糠90
gをよく混合し、水道水にて水分含有率65%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い直径1cmの穴をあ
けた後、キャップで打栓したオガクズ固型培養基を12
0℃、60分間高圧蒸気滅菌したものをA培地とした。
また該培地調製方法に準じ、コーンコブミール130
g、マメカワ60g、フスマ30gをよく混合して同様
に作成したものをB培地、針葉樹オガクズ50g、綿実
殻粉砕物50g、米糠100gをよく混合して同様に作
成したものをC培地、針葉樹オガクズ100g、マメカ
ワ70gをよく混合して同様に作成したものをD培地と
し、それぞれ用いた。各使用培地で得られた子実体の収
量(g/ビン)、官能検査の結果を表1に示す。The present invention will be described in detail below. The lyophilum ulmarium strain is disclosed in JP-A-63-2734.
Riofilum ulmarium Lu described in Japanese Patent Publication No. 67
1-2 (FERM P-12584), Riofilum ulmarium Lu 1-8 (FERM BP-14
16), Riofilum Ulmarium Lu 1-17
(FERM BP-1417) and riophyllum ulmarium M-8171 (FERM BP-1415) are used, artificial cultivation of each strain is carried out by the method described in the publication, and fruit bodies at an appropriate harvest time are harvested to obtain fruit bodies. A sensory test was performed. As the medium for artificial cultivation, 50 g of coniferous sawdust, 50 g of broadleaf sawdust and 90 g of rice bran described in the publication are used.
After mixing well with g and adjusting the water content to 65% with tap water, it was pressed into a polypropylene 850 ml wide-mouthed bottle and a hole with a diameter of 1 cm facing downward from the center of the bottle mouth was opened. 12 pieces of sawdust solid culture medium capped with a cap
What was subjected to high-pressure steam sterilization at 0 ° C. for 60 minutes was used as the A medium.
According to the method for preparing the medium, corncob meal 130
g, Mamekawa 60g, bran 30g were mixed in the same way, and B medium, 50g coniferous sawdust 50g, crushed cottonseed shell 50g, and 100g rice bran were similarly mixed to produce C medium, coniferous sawdust. A medium prepared in the same manner by thoroughly mixing 100 g and 70 g of Memekawa was used as D medium. Table 1 shows the yield (g / bin) of fruiting bodies and the result of the sensory test obtained in each medium used.
【0007】[0007]
【表1】 表 1 ──────────────────────────────────── 使 用 菌 株 培 地 収量(g/ビン) 官能検査 ──────────────────────────────────── リオフィラム ウルマリウム A 110 − Lu 1−2 B 178 + C 140 + D 153 + リオフィラム ウルマリウム A 112 ++ Lu 1−8 B 179 +++ C 142 +++ D 150 +++ リオフィラム ウルマリウム A 108 − Lu 1−17 B 180 ++ C 144 + D 155 + リオフィラム ウルマリウム A 113 − M−8171 B 188 +++ C 157 +++ D 171 +++ ────────────────────────────────────[Table 1] Table 1 ──────────────────────────────────── Bacteria used Cultivated land Yield (g / bottle) Sensory test ──────────────────────────────────── Riofilum Ulmarium A 110- Lu 1-2 B 178 + C 140 + D 153 + Riophyllum Ulmarium A 112 ++ Lu 1-8 B 179 +++ C 142 +++ D 150 +++ Riofilum Ulmarium A 108- Lu 1-17 B 180 ++ C 144 + D 155 + Riofilum Ulmarium A 113- M-8171 B 188 +++ C 157 +++ D 171 +++ ────────────────────────────────────
【0008】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5m
l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。In the sensory test, lyophilum ulmarium fruit bodies are stir-fried in oil, and quinine sulfate is used as a standard substance for bitterness by a panelist. Fried oil is performed as follows. That is, the base of the obtained fruit body of Riofilum ulmarium is removed, divided into individual pieces, lightly washed with water, and well drained. Next, add 5m of salad oil to the frying pan.
Add 1 liter, spread all over and heat oil well over medium heat. Riofilum ulmarium fruiting body 100g (wet weight)
Add the ingredients to the pan and stir with chopsticks for 2 minutes or more, and heat the whole mixture. When the pan is dry, turn off the heat. Allow the fruiting body to cool to room temperature and not bitter the panelist,
The four levels of bitterness, pretty bitterness, and remarkably bitterness are evaluated.
In the following, − in each table means no bitterness, + means bitterness, ++ means much bitterness, and ++ means remarkably bitterness. This +
The middle of − is the threshold of bitterness with quinine sulfate as a standard substance, which is the bitterness of Riofilum ulmarium fruiting body, that is,
It is a bitterness with almost no bitterness, the threshold value of quinine sulfate as a standard substance, 0.000008M (Shizuyuki Ota, "Knowledge of food seasoning" 2nd edition, 1st edition, October 1, 1985,
Corresponds to the bitterness of Koshou Shobo, page 38). Among the above-mentioned test strains, riophyllum ulmarium Lu 1-2 is the strain with the lowest bitterness, but when B, C or D medium is used, the yield increases but the fruiting body obtained has a bitter taste. .
【0009】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。[0009] In the present invention the high bitterness of medium Lyophyllum ulmarium bitterness of fruit bodies obtained when performing the artificial cultivation of Lu 1-2 are means + or more media by sensory test, the upper Symbol B, C , D medium. Further, the new strain of the present invention refers to a strain in which the bitterness of fruiting bodies obtained by cultivating the strain in a high-bittering medium is such that the bitterness is hardly felt in at least one high-bittering medium.
【0010】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11
2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。Next, breeding of the new strain of the present invention will be described. 1. Selective breeding Riofilum ulmarium 11 occurring in the natural world
Tissue separation was carried out from the two fruiting bodies to obtain a purely separated mycelium 90 strain. Next, this 90 strain was subjected to a cultivation test using the B medium. By a cultivation test, fruit bodies of 48 strains that formed fruit 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 a cultivation test in B, C and D media, and Riofilum ulmarium Lu 1-13 was selected as a strain having the lowest bitterness and suitable for industrial cultivation.
【0011】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、老時中央にき製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。The riophyllum ulmarium will be described below. Riofilum Ulmarium Lu 1
The -13 strain was isolated by the present inventors from a fruit body that had been crowded on a dead tree in Urabandai, Fukushima Prefecture, and the characteristics of the fruit body and spores are as follows. Fruit bodies are crowded, and umbrellas are diameter 5
~ 1.5 cm, round or irregularly shaped round mountain, smooth surface,
It has a moist, white to brownish cream color, often presents a slightly dark mottle, and may cause squeezing in the middle of old age.
The flesh is white and has a wide variety of patterns. The handle is eccentric and curved,
3 to 7 x 1 to 2 cm, almost the same color as the bulk, white at the top, fluffy or powdery. Spores are almost spherical, smooth, colorless,
4.5 to 5.5 × 3.5 to 4.5 μm, and the pattern was white. Or more of the features of the Seiya Ito al., "Japan fungi magazine" the second volume fifth issue (195 nine years, nourishing Kashikodo publication) when compared with the description of, this bacterium is clearly to be a Lyophyllum ulmarium.
【0012】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。1
7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)60
ml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。Next, Riofilum ulmarium Lu
The various bacteriological properties of strains 1-13 are shown below. (1) Malt extract agar medium (25 ° C culture) Day 7: vigorous growth. The colony diameter is 40 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 10: Mycelia grow on the entire petri dish. Day 17: Dense aerial mycelia develop on the entire surface. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 7: vigorous growth. The colony diameter is 36 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 10: Mycelia grow on the entire petri dish. Day 17: Dense aerial mycelia develop on the entire surface. Light yellow in the center of the colony, white in others. (3) Czapek-Dox agar medium (25 ° C. culture) Day 7: Small growth. The colony diameter is 26 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 17: Mycelia grow on the entire petri dish. Mycelium is dendritic, thin and white. (4) Sabouraud agar medium (25 ° C. culture) Day 7: vigorous growth. The colony diameter is 42 mm. White, cotton-like dense hyphae, aerial hyphae. Day 10: Mycelia grow on the entire petri dish. An abundant amount of aerial mycelia is produced, and the hyphae are cotton-like and white. (5) Oatmeal agar medium (25 ° C. culture) Day 7: Vigorous growth. The colony diameter is 36 mm. Hyphae are well branched and elongated, and aerial hyphae are few. Day 10: Mycelia grow on the entire petri dish. A large amount of cotton-like aerial hyphae are produced. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 7: Small growth. The colony diameter is 21 mm. The hyphae are white and linearly elongated, forming radial colonies. 1
Day 7: Mycelia grow on the entire dish. A large amount of aerial hyphae are produced. Mycelia are white. (7) YpSs agar medium (25 ° C. culture) Day 7: vigorous growth. The colony diameter is 40 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Matte shape. Day 10: Mycelia grow on the entire petri dish. A large amount of dense aerial hyphae are produced. The mycelium is white, but the medium turns yellow. (8) Phenol oxidase assay medium (25 ° C. culture) Day 7: Small growth. The colony diameter is 18 mm. Mycelium is white and grows short and mat-like with few aerial mycelia. The medium is browned and the radius of browning is 40 mm. Day 17: Medium growth. The colony diameter is 37 mm. The browning radius is 42 mm. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 7-8. Also,
The viable pH range was pH 3.5 to 10.
【0013】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−817
1、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−)。Next, Riofilum ulmarium Lu
Based on the taxonomic fact that the hyphae are different hyphae, if the sexes possessed by both hyphae are different from each other, the sex Differences were examined by inversion culture on agar. The tested Riofilum ulmarium is Riofilum ulmarium IFO 9637, Riofilum
Ulmarium IFO 30525, Riofilum Ulmarium IFO 30775, Riofilum Ulmarium Lu 1-2, Riofilum Ulmarium Lu 1-8, Riofilum Ulmarium Lu
1-17, Riofilum Ulmarium M-817
1. Riofilum ulmarium SA. It should be noted that Riofilum ulmarium SA is an isolated strain from the commercially available Riofilum ulmarium fruiting body. Each of the above-mentioned binuclear hyphae of riophyllum ulmarium was cut out from the preserved slant as a block of 3 × 3 × 3 mm, and each was inoculated in the center of the PGY agar medium so as to confront the binuclear hyphae of riophyllum ulmarium Lu 1-13. After culturing (at 2 cm intervals) at 25 ° C. for 14 days, it was determined whether a band line was formed between colonies of both strains. The results are shown in Table 2. (+ If banding occurred, -if not).
【0014】[0014]
【表2】 表 2 ───────────────────────────────── リオフィラム ウルマリウム IFO 9637 + IFO 30525 + IFO 30775 + Lu 1−2 + Lu 1−8 + Lu 1−17 + M−8171 + SA + ─────────────────────────────────[Table 2] Table 2 ────────────────────────────────── Riophyllum Ulmarium IFO 9637 + IFO 30525 + IFO 30775 + Lu 1-2 + Lu 1-8 + Lu 1-17 + M-8171 + SA + ──────────────────────────────────
【0015】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 Lu1−13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 L
u 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。As can be seen from Table 2, all of the above-mentioned strains produced zonal lines in the confrontation culture with Riophyllum ulmarium Lu1-13, and it is clear that Riofilum ulmarium Lu1-13 is a new strain. . As described above, as a new strain by selective breeding of the present invention, for example, Riofilum ulmarium L
u 1-13 can be mentioned, but the bitterness of the fruiting body obtained by cultivating in a high bittering medium as in the case of the strain is such that, for at least one type of high bittering medium, the bitterness is less than or equal to a level at which little bitterness is felt. The strains shown in are all belonging to the present invention.
【0016】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63−2
73467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−81
71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165
3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−042
9、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−68
04、リオフィラム ウルマリウム K−6806とそ
れぞれ命名した。2. Cross Breeding The new strain of the invention was obtained by cross breeding. JP-A-63-2
As described in Japanese Patent Publication No. 73467, by crossing Riofilum ulmarium Lu 1-8 with Riofilum ulmarium Lu 1-17, Riofilum ulmarium M-81 excellent in yield, growth rate and fruiting body morphology.
71 has been obtained. In order to cross-bred strains having excellent cultivation characteristics and reduced bitterness of fruiting bodies, this riophyllum ulmarium lu 1-8, riophyllum
Both strains of Ulmarium Lu 1-17 were used as parent strains. Riofilum ulmarium Lu 1-8 and riophyllum ulmarium Lu 1-17 were grown in A medium, respectively, and fruiting bodies were generated by a normal operation. Spores were collected from each of the fruiting bodies, germinated mononuclear hyphae were isolated, and 57 strains were bred for 165 rounds.
3 strains were obtained. This 1653 strain was subjected to a cultivation test using B medium, and 80 strains with low bitterness were selected by a sensory test,
Next, a cultivation test was carried out in B, C, and D media, and the cultivation characteristics were excellent, and the bitterness was at least one of the high-bitterness media
Eight strains having almost no bitterness were selected, and lyophilum ulmarium K-042 was selected.
9, Riofilum ulmarium K-0207, Riofilum ulmarium K-0202, Riofilum ulmarium K-0259, Riofilum ulmarium K-1004, Riofilum ulmarium
K-1257, Riofilum ulmarium K-68
04, and Riofilum ulmarium K-6806, respectively.
【0017】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64m
m。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)60
ml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。Next, this Riofilum ulmarium K
The bacteriological properties of the -0429 strain are shown. (1) Malt extract agar medium (25 ° C. culture) Day 5: Colony diameter 18 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 10: Colony diameter 47 mm. 20
Day: Colony diameter is 83 mm. It grows radially and produces dense aerial hyphae on the entire surface. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 40 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: Colony diameter 55 mm. Day 20: Colony diameter 66 mm. Dense aerial hyphae are produced on the entire surface. Cotton-like colony with white hyphae. (3) Czapek-Dox agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 28 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 20:
The colony diameter is 52 mm. Mycelium is dendritic, thin and white. (4) Sabouraud agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 50 mm. White, cotton-like dense hyphae, aerial hyphae. Day 20: Mycelia grow on the entire dish. It grows radially and produces aerial mycelia on the entire surface. Mycelia are white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 51 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow on the entire dish. A flocculent colony with a large amount of aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 27 mm. The hyphae are white and form dendritic colonies. 20th day: Colony diameter is 48 mm. Produces aerial mycelia. Dendritic colony with white hyphae. (7) YpSs agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 49 mm. Fluffy colonies produce white hyphae and aerial hyphae. Day 20: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 5: Small growth. The colony diameter is 18 mm. Mycelium is white and produces a large amount of aerial mycelium. The medium turned brown and the browning radius was 34 mm. Day 15: Strong growth. Colony diameter is 64m
m. The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 7-8. Also,
The viable pH range was pH 3.5 to 10.
【0018】次にリオフィラム ウルマリウム K−0
207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)60
ml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。Next, Riofilum ulmarium K-0
The mycological properties of strain 207 are shown. (1) Malt extract agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 61 mm. White, radial colonies, small amount of aerial mycelia. Day 20: Mycelia grow on the entire dish. There are few aerial hyphae. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Vigorous growth. The colony diameter is 53 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface.
The colony is cotton and white. (3) Czapek-Dox agar medium (25 ° C. culture) Day 7: vigorous growth. The colony diameter is 51 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 15: Colony diameter 75 mm. Mycelium is dendritic, thin and white. Day 20:
The colony diameter is 77 mm. (4) Sabouraud agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 58 mm. White, radial dense hyphae, aerial hyphae. Day 15: Mycelia grow on the entire dish. Aerial hyphae are produced, and the hyphae are radial and white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 66 mm. Colonies are flocculent and produce aerial hyphae. Day 15: Mycelia grow on the entire dish. Flocky colonies produce aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 33 mm. Hyphae are white and slightly thin, dendritic, with aerial hyphae. 20
Day: Colony diameter is 74 mm. The colony is thin and dendritic,
Mycelia are white. (7) YpSs agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 65 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 63 mm. Mycelium is white and produces a large amount of aerial mycelia. The medium is browned and the radius of browning is 65 mm. Day 15: Mycelia grow on the entire dish. A large amount of aerial mycelia are produced, and the entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 7-8. Also,
The viable pH range was pH 3.5 to 10.
【0019】次にリオフィラム ウルマリウム K−0
202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)60
ml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。Next, Riofilum ulmarium K-0
The mycological properties of the 202 strain are shown. (1) Malt extract agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 60 mm. White, radial colonies, small amount of aerial mycelia. Day 20: Mycelia grow on the entire dish. There are few aerial hyphae. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Vigorous growth. The colony diameter is 59 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface.
The colony is cotton and white. (3) Czapek-Dox agar medium (25 ° C. culture) Day 5: vigorous growth. The colony diameter is 16 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 15: Colony diameter is 62 mm. Mycelium is dendritic, thin and white. Day 20:
The colony diameter is 77 mm. (4) Sabouraud agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 66 mm. White, radial dense hyphae, aerial hyphae. Day 15: Mycelia grow on the entire dish. Aerial hyphae are produced, and the hyphae are radial and white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 68 mm. Colonies are flocculent and produce aerial hyphae. Day 15: Mycelia grow on the entire dish. Flocky colonies produce aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 33 mm. Hyphae are white and slightly thin, dendritic, with aerial hyphae. 20
Day: Colony diameter is 62 mm. The colony is thin and dendritic,
Mycelia are white. (7) YpSs agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 68 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 61 mm. Mycelium is white and produces a large amount of aerial mycelia. The medium turned brown. Day 15:
Mycelia grow on the entire dish. A large amount of aerial hyphae,
The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, 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.
【0020】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)60
ml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。Next, this Riofilum ulmarium K
The mycological properties of the -0259 strain are shown. (1) Malt extract agar medium (25 ° C. culture) Day 5: Colony diameter 24 mm. A large amount of white and dense mycelia and aerial mycelia are produced. Day 10: Colony diameter 64 mm. Day 15: Mycelia grow on the entire dish. It grows radially and produces dense aerial hyphae on the entire surface. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 61 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: 85 mm colony diameter. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface. Cotton-like colony with white hyphae. (3) Czapek-Dox agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 48 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 20:
The colony diameter is 75 mm. Mycelium is dendritic, thin and white. (4) Sabouraud agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 65 mm. White, cotton-like dense hyphae, aerial hyphae. Day 20: Mycelia grow on the entire dish. It grows radially and produces aerial mycelia on the entire surface. Mycelia are white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 67 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow on the entire dish. A flocculent colony with a large amount of aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 39 mm. The hyphae are white and form dendritic colonies. Day 20: Colony diameter 66 mm. Produces aerial mycelia. Dendritic colony with white hyphae. (7) YpSs agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 63 mm. Fluffy colonies produce white hyphae and aerial hyphae. Day 20: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 5: Small growth. The colony diameter is 29 mm. Mycelium is white and produces a large amount of aerial mycelium. The medium turned brown. Day 15: Strong growth. The colony diameter is 88 mm. The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Also,
The viable pH range was pH 3.5 to 10.
【0021】次にリオフィラム ウルマリウム K−1
004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)60
ml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。Next, Riofilum ulmarium K-1
The mycological properties of strain 004 are shown below. (1) Malt extract agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 64 mm. White, radial colonies, small amount of aerial mycelia. Day 20: Mycelia grow on the entire dish. There are few aerial hyphae. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Vigorous growth. The colony diameter is 64 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface.
The colony is cotton and white. (3) Czapek-Dox agar medium (25 ° C. culture) Day 5: vigorous growth. The colony diameter is 14 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 15: Colony diameter 60 mm. Mycelium is dendritic, thin and white. Day 20:
The colony diameter is 75 mm. (4) Sabouraud agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 83 mm. White, radial dense hyphae, aerial hyphae. Day 15: Mycelia grow on the entire dish. Aerial hyphae are produced, and the hyphae are radial and white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 81 mm. The colonies are flocculent and produce large numbers of aerial hyphae. Day 15: Mycelia grow on the entire dish. Flocky colonies produce aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 23 mm. Hyphae are white and slightly thin, dendritic, and accompanied by aerial hyphae. 20
Day: Colony diameter is 45 mm. The colony is thin and dendritic.
Mycelia are white. (7) YpSs agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 80 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 68 mm. Mycelium is white and produces a large amount of aerial mycelia. The medium turned brown. Day 15:
Mycelia grow on the entire dish. A large amount of aerial hyphae,
The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-7. Also,
The viable pH range was pH 3.5 to 10.
【0022】次にリオフィラム ウルマリウム K−1
257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)60
ml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。Next, Riofilum ulmarium K-1
The various bacteriological properties of strain 257 are shown. (1) Malt extract agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 71 mm. White, radial colonies, small amount of aerial mycelia. Day 20: Mycelia grow on the entire dish. There are few aerial hyphae. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Vigorous growth. The colony diameter is 66 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface.
The colony is cotton and white. (3) Czapek-Dox agar medium (25 ° C. culture) Day 5: vigorous growth. The colony diameter is 26 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 15: Colony diameter is 71 mm. Mycelium is dendritic, thin and white. Day 20:
The colony diameter is 81 mm. (4) Sabouraud agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 79 mm. White, radial dense hyphae, aerial hyphae. Day 15: Mycelia grow on the entire dish. Aerial hyphae are produced, and the hyphae are radial and white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 79 mm. The colonies are flocculent and produce large numbers of aerial hyphae. Day 15: Mycelia grow on the entire dish. Flocky colonies produce aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 37 mm. Hyphae are white and slightly thin, dendritic, with aerial hyphae. 20
Day: Colony diameter is 68 mm. The colony is thin and dendritic.
Mycelia are white. (7) YpSs agar medium (25 ° C. culture) Day 10: Very vigorous growth. The colony diameter is 64 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 70 mm. Mycelium is white and produces a large amount of aerial mycelia. The medium turned brown. Day 15:
Mycelia grow on the entire dish. A large amount of aerial hyphae,
The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Also,
The viable pH range was pH 3.5 to 10.
【0023】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)60
ml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。Next, this Riofilum ulmarium K
The mycological properties of the -6804 strain are shown. (1) Malt extract agar medium (25 ° C. culture) Day 5: colony diameter 25 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 10: Colony diameter is 61 mm. 15
Day: Colony diameter is 83 mm. It grows radially and produces dense aerial hyphae on the entire surface. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 61 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: colony diameter 86 mm. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface. Cotton-like colony with white hyphae. (3) Czapek-Dox agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 49 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 20:
The colony diameter is 76 mm. Mycelium is dendritic, thin and white. (4) Sabouraud agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 70 mm. White, cotton-like dense hyphae, aerial hyphae. Day 20: Mycelia grow on the entire dish. It grows radially and produces aerial mycelia on the entire surface. Mycelia are white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 74 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow on the entire dish. A flocculent colony with a large amount of aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 29 mm. The hyphae are white and form dendritic colonies. Day 20: Colony diameter 53 mm. Produces aerial mycelia. Dendritic colony with white hyphae. (7) YpSs agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 72 mm. Fluffy colonies produce white hyphae and aerial hyphae. Day 20: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 5: Small growth. The colony diameter is 28 mm. Mycelium is white and produces a large amount of aerial mycelium. The medium turned brown. Day 15: Mycelia grow on the entire dish. The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Also,
The viable pH range was pH 3.5 to 10.
【0024】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)60
ml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。Next, this Riofilum ulmarium K
The various bacteriological properties of the −6806 strain are shown. (1) Malt extract agar medium (25 ° C. culture) Day 5: Colony diameter 19 mm. A large amount of white and dense mycelia and aerial mycelia are produced. Day 10: Colony diameter 59 mm. Day 15: colony diameter 81 mm. It grows radially and produces dense aerial hyphae on the entire surface. Mycelia are white. (2) Potato-glucose agar medium (25 ° C. culture) Day 10: Medium growth. The colony diameter is 55 mm. A large amount of white, dense hyphae and aerial hyphae are produced. Day 15: colony diameter 80 mm. Day 20: Mycelia grow on the entire dish. Dense aerial hyphae are produced on the entire surface. Cotton-like colony with white hyphae. (3) Czapek-Dox agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 38 mm. Very few mycelia and aerial hyphae that grow in a dendritic form and are extremely thin. Day 20:
The colony diameter is 66 mm. Mycelium is dendritic, thin and white. (4) Sabouraud agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 66 mm. White, cotton-like dense hyphae, aerial hyphae. Day 20: Mycelia grow on the entire dish. It grows radially and produces aerial hyphae on the entire surface. Mycelia are white. (5) Oatmeal agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 71 mm. A flocculent colony that produces aerial hyphae. Day 20: Mycelia grow on the entire dish. A flocculent colony with a large amount of aerial hyphae. Mycelia are white. (6) Synthetic Mucor agar medium (25 ° C. culture) Day 10: Small growth. The colony diameter is 31 mm. The hyphae are white and form dendritic colonies. Day 20: Colony diameter is 61 mm. Produces aerial mycelia. Dendritic colony with white hyphae. (7) YpSs agar medium (25 ° C. culture) Day 10: vigorous growth. The colony diameter is 70 mm. Fluffy colonies produce white hyphae and aerial hyphae. Day 20: Mycelia grow on the entire dish. A large amount of dense aerial hyphae are produced. Mycelia are white. (8) Phenol oxidase assay medium (25 ° C. culture) Day 5: Small growth. The colony diameter is 28 mm. Mycelium is white and produces a large amount of aerial mycelium. The medium turned brown. Day 15: Strong growth. The colony diameter is 86 mm. The entire surface of the medium turns brown. The growth rate significantly varies depending on the old and new seeds. (9) Optimal growth temperature PGY agar medium was inoculated with a discoid inoculum having a diameter of 5 mm, 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. Also, 5
Almost no growth occurred at 35 ° C and 35 ° C. (10) Optimal growth pH PGY liquid medium (PGY agar without agar) 60
Dispense each 100 ml into a 100 ml Erlenmeyer flask to sterilize, adjust each pH with acid or alkali, and inoculate inoculum, then
After static culture at ℃ for 15 days, the dry weight of the cells was measured. As a result, the optimum growth pH was 6-8. Also,
The viable pH range was pH 3.5 to 10.
【0025】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−020
7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−817
1、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。Next, Riofilum ulmarium K-
0429 strain, riophyllum ulmarium K-020
7 strains, Riofilum ulmarium K-022 strain,
Riofilum ulmarium K-0259 strain, riophyllum ulmarium K-1004 strain, riophyllum ulmarium K-1257 strain, riophyllum ulmarium K-6804 strain and riophyllum ulmarium K-6806 strain and liophyllum ulmarium sexuality possessed by both hyphae. Based on the taxonomic fact that the hyphae are different hyphae if the factors are different from each other, the sex factors were examined by counterculture on an agar medium. The tested Riofilum ulmarium is Riofilum ulmarium IFO 9637, Riofilum
Ulmarium IFO 30525, Riofilum Ulmarium IFO 30775, Riofilum Ulmarium Lu 1-2, Riofilum Ulmarium Lu 1-8, Riofilum Ulmarium Lu
1-17, Riofilum Ulmarium M-817
1. Riofilum ulmarium SA and Riofilum ulmarium Lu 1-13.
【0026】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−68
04株、又はリオフィラム ウルマリウム K−680
6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−)。Each of the above-mentioned binuclear hyphae of Riofilum ulmarium was cut out from the preserved slant as a block of 3 × 3 × 3 mm, and each of them was placed in the central portion of the PGY agar medium, and Riofilum ulmarium K-0429 strain, Riofilum ulmarium K-0207 strain, Riofilum ulmarium K-0202 strain, riophyllum ulmarium K-0259 strain, riophyllum ulmarium K-1004 strain, riophyllum ulmarium K
-1257 strain, Riofilum ulmarium K-68
04 strain, or Riofilum ulmarium K-680
Inoculate with 6 strains of dinuclear hyphae (2 cm intervals), 25
After culturing at 14 ° 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. (+ If banding occurred, -if not).
【0027】[0027]
【表3】 [Table 3]
【0028】[0028]
【表4】 [Table 4]
【0029】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−02
59、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。As can be seen from Tables 3 and 4, Riofilum ulmarium K-0429, Riophyllum ulmarium K-0207, Riofilum ulmarium
K-0202, Riofilum ulmarium K-02
59, Riofilum ulmarium K-1004, Riofilum ulmarium K-1257, Riofilum ulmarium K-6804, and Riofilum.
Urmalium K-6806 all produced zoning in opposite cultures with other strains, respectively, which makes it clear that both strains are new strains.
【0030】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−020
2、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。As the new strains bred by the crossing of the present invention as described above, for example, Riofilum ulmarium K-0429 and Riofilum ulmarium K-.
0207, Riofilum Ulmarium K-020
2, Riofilum ulmarium K-0259, Riofilum ulmarium K-1004, Riofilum ulmarium K-1257, Riofilum ulmarium K-6804, Riofilum ulmarium K
-6806 and the like, but the bitterness of the fruiting body obtained by cultivating in a high bittering medium as in the case of the strain is such that, for at least one type of high bittering medium, the bitterness is less than or equal to a level at which little bitterness is felt. All the strains shown belong to the present invention.
【0031】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。As described above, the new strain according to the present invention has a bitterness of fruiting bodies which is less than or equal to the bitterness of at least one type of high bittering medium even when grown in a high bittering medium during artificial cultivation. It has the characteristic that it is stable regardless of the type of medium and has little bitterness in fruiting bodies. Table 5 and Table 6 show the bitterness and yield of fruiting bodies at the proper harvesting time, which were obtained by artificial cultivation using the new strains A, B, C and D of the present invention.
Shown in. The above mentioned Riophyllum ulmarium SA
The results are also shown in Table 6 together.
【0032】[0032]
【表5】 表 5 ──────────────────────────────────── リオフィラム ウルマリウム 培 地 収量(g/ビン) 官能検査 ──────────────────────────────────── Lu 1−13 A 105 − B 168 − C 143 − D 157 − K−0429 A 112 − B 184 − C 157 − D 164 − K−0207 A 111 − B 188 − C 161 − D 178 − K−0202 A 98 − B 187 − C 154 − D 162 − K−0259 A 108 − B 192 − C 162 − D 165 − ────────────────────────────────────[Table 5] Table 5 ──────────────────────────────────── Riofilum Ulmarium Cultivated Yield (g / bottle) Sensory test ──────────────────────────────────── Lu 1-13 A 105 − B 168- C143- D157- K-0429 A 112- B184- C157- D164- K-0207 A 111- B 188- C 161- D178- K-022 A 98- B187- C154- D 162- K-0259 A 108- B 192- C 162 − D165- ────────────────────────────────────
【0033】[0033]
【表6】 表 6 ──────────────────────────────────── リオフィラム ウルマリウム 培 地 収量(g/ビン) 官能検査 ──────────────────────────────────── K−1004 A 99 − B 188 − C 158 − D 161 − K−1257 A 109 − B 186 − C 160 − D 157 − K−6804 A 120 − B 195 − C 163 − D 171 − K−6806 A 116 − B 191 − C 162 − D 165 − SA A 114 + B 189 +++ C 155 ++ D 168 +++ ────────────────────────────────────[Table 6] Table 6 ──────────────────────────────────── Riofilum Ulmarium Cultivated Yield (g / bottle) Sensory test ──────────────────────────────────── K-1004 A 99- B 188- C158- D 161- K-1257 A 109- B186- C 160- D157- K-6804 A 120- B195- C163- D 171- K-6806 A 116- B 191- C 162 − D165- SA A 114 + B 189 +++ C 155 ++ D 168 +++ ────────────────────────────────────
【0034】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−020
2、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。As shown in Tables 5 and 6, Riofilum ulmarium Lu 1-13, Riofilum ulmarium K-0429, Riofilum ulmarium K
-0207, Riofilum ulmarium K-020
2, Riophyllum ulmarium K-0259, Riofilum ulmarium K-1004, Riofilum ulmarium K-1257, Riofilum ulmarium K-6804, Riofilum ulmarium
Each of the K-6806 strains does not show bitterness in the conventional artificially cultivated strains even when a medium in which fruiting bodies at the proper harvesting time exhibit bitterness is used, and thus the fruiting body yield is increased but the bitterness is also increased. Even with a high bittering medium that was difficult to use, fruiting bodies without bitterness can be stably obtained in high yield.
【0035】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。The mycelium produced by inoculating the above strain into a medium does not show bitterness, and is suitable for use in food. It should be noted that lyophilium ulmarium mycelium has a large amount of dietary fiber and is known to have an anti-cancer effect, and foods using the mycelium are particularly useful for maintaining health.
【0036】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−025
9、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−042
9、 Lyophyllumulmarium K−0207、 Lyophyllum
ulmarium K−0202、 Lyophyllumulmarium K−
0259、 Lyophyllum ulmarium K−1004、 Lyo
phyllumulmarium K−1257、 Lyophyllum ulmariu
m K−6804、 Lyophyllumulmarium 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−1257
1)、微工研菌寄第12570号(FERM P−12
570)、微工研菌寄第12569号(FERMP−1
2569)、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(FE
RM P−12982)、微工研菌寄第12983号
(FERMP−12983)、微工研菌寄第12984
号(FERM P−12984)、微工研菌寄第129
85号(FERM P−12985)として寄託されて
いる。New strains of the strains bred in the present invention, Riofilum ulmarium Lu 1-13, Riofilum
Ulmarium K-0429, Riofilum Ulmarium K-0207, Riofilum Ulmarium K-
0202, Riofilum ulmarium K-025
9. Lyophyllum ulmarium K-1004, Lyophyllum ulmarium K-1257, Riofilum ulmarium K-6804, and Lyophyllum ulmarium K-6806 are Lyophyllum ulmarium, respectively.
Lu 1-13, Lyophyllum ulmarium K-042
9, Lyophyllumulmarium K-0207, Lyophyllum
ulmarium K-0202, Lyophyllum ulmarium K-
0259, Lyophyllum ulmarium K-1004, Lyo
phyllumulmarium K-1257, Lyophyllum ulmariu
m K-6804, Lyophyllumulmarium K-6806
Is displayed in the Institute of Microbial Technology, National Institute of Advanced Industrial Science and Technology, respectively.
1), Microtechnology Research Institute, No. 12570 (FERM P-12
570), Microtechnology Research Institute, No. 12569 (FERMP-1
2569), Microtech Lab, No. 12980 (FERM P
-12980), and Microtechnology Research Institute, No. 12981 (FERM
P-12981), Microtechnology Research Institute, No. 12982 (FE)
RM P-12982), Microindustry Research Institute No. 12983 (FERMP-12983), Microindustry Research Institute No. 12984.
No. (FERM P-12984), Microcosm Research Institute 129
Deposited as No. 85 (FERM P-12985).
【0037】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 F
ine 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( Mer
ck )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UV210
nm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。The bitterness component in the fruiting body of riophyllum ulmarium can also be quantified as follows. Riofilum ulmarium Edible part in fruiting body (kasa part,
The handle) is freeze-dried, and this is crushed to obtain a fruit-body freeze-dried powder. 0.5 g (dry weight) of this lyophilized powder was extracted with 35 ml of ethyl acetate by shaking at 25 ° C. for 24 hours, filtered, concentrated under reduced pressure, and dissolved in 1 ml of methanol while heating to obtain an extracted sample. FPLC from the above sample 200 μl
[Pharmacia F
ine Chemicals)] and collect the bitter component fraction as follows. Column is Lober RP-8 [Merck (Mer
CK), φ1.0 × 24 cm], solvent 85% methanol (Kanto Chemical; for liquid chromatography), 1.0 ml / min, room temperature, detection wavelength 210 nm, elution time 10-45
Collect fractions of minutes. Min toga fraction was again concentrated under reduced pressure, methanol: ethanol: water = 3: 4: was dissolved in 4 solvent 1.0 ml, the 50μl by HPLC [manufactured by Shimadzu Corporation LC-6A System] The bitterness component fraction is collected.
The column is μ Bondapak C 18 [Waters (Waters
) Manufactured by using φ0.39 × 30cm], methanol: ethanol: water = 3: 4: 4 in a solvent (Kanto Kagaku; liquid chromatography for methanol, Nacalai Tesque; special grade ethanol), 0.7 ml / min, Temperature 40 ℃, detection wavelength UV210
Under the condition of nm, the dry weight of the fraction having an elution time of 32.5 to 44.0 minutes is measured. The component eluting in this fraction (hereinafter abbreviated as Fraction A) exhibits a strong bitterness in the sensory test, and the bitterness of Riofilum ulmarium fruiting body is
caused by.
【0038】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。The cotton seed shells used in the medium of the present invention are
The present invention has been found by the present inventors to be a substrate that can be obtained at low cost and that significantly increases the yield of mushrooms and is useful for artificial cultivation of mushrooms. When this cottonseed shell is used in a medium, it may be mixed with another medium base material or used alone. 15% of cottonseed husks based on dry weight of medium
When used at -60%, the effect of increasing the yield is the best, but the amount used is not limited to this. Mushrooms that can be cultivated in good yield using this cottonseed shell as a medium include, for example, liophyllum ulmarium of the present invention, shiitake mushroom,
There are oyster mushrooms, enoki mushrooms, maitake mushrooms, and the like. For example, strains of these mushrooms may be used for ordinary artificial cultivation.
【0039】[0039]
【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。 EXAMPLES Examples of artificial cultivation of a new strain of ryophilum ulmarium according to the present invention are shown below, but the present invention is not limited to the scope of the following examples. In addition, below
Examples 1 to 3 described above are listed as reference examples of the present invention.
It is an example.
【0040】実施例1
液体PGY培地(寒天を含まないPGY寒天培地)10
0ml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、2
5℃、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。Example 1 Liquid PGY medium (PGY agar containing no agar) 10
To 0 ml, Riofilum ulmarium Lu 1-13
(FERM P-12571) and inoculated at 25 ° C for 10
Liquid culture was obtained by culturing for a day. On the other hand, 130 g of corn cob meal, 60 g of mamekawa and 30 g of bran were mixed well and the water content was adjusted to 62% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was subjected to high-pressure steam sterilization at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with 20 ml of the liquid inoculum. Dark place, 2
When the culture medium was cultured for 35 days under the conditions of 5 ° C. and a humidity of 50 to 60%, hyphae were spread on the entire bottle. Further, the culture was continued for 40 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply sufficient water, the excess tap water was discarded.
9 ° C, humidity 90-95%, illuminance 20lux 9
Continue culturing for a day to obtain fruit body primordia, and further illuminance to 200
The seeds were picked up after being cultivated for 15 days and cultured for 13 days.
The yield of the fruiting body thus obtained was 168 g, and no bitterness was felt in the sensory test.
【0041】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。The sensory test of fruiting bodies was conducted as follows. The base of the handle of the fruiting body obtained by the above bottle cultivation was removed, and the fruit body was divided into pieces one by one, lightly washed with water, and well drained. On the other hand, add 5 ml of salad oil to a frying pan, spread it over the whole, heat the oil sufficiently with medium heat, add 100 g (wet weight) of fruiting body, cook for 2 minutes or more with stirring with chopsticks, and cook in the frying pan. Stir until it is dry, then turn off the heat. The fruit body fried in oil was cooled to room temperature, and quinine sulfate was used as a standard substance, and the bitterness was judged by 10 panelists.
【0042】実施例2
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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、2
5℃、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。Example 2 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium Lu 1-13. Dark place, 2
When the culture medium was cultured for 30 days under the conditions of 5 ° C. and a humidity of 50 to 60%, hyphae spread to the entire bottle. Further, the culture was continued for 35 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply sufficient water, the excess tap water was discarded.
9 ° C, humidity 90-95%, illuminance 20lux 9
Continue culturing for a day to obtain fruit body primordia, and further illuminance to 200
The seeds were picked up after being cultivated for 15 days and cultured for 13 days.
The yield of the fruiting body thus obtained was 143 g, and no bitterness was felt in the sensory test of the fruiting body.
【0043】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1
3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。Example 3 100 g of softwood sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C for 60 minutes, cooled to room temperature, and then lyophilum ulmarium Lu 1-
Inoculated with 13 solid inoculums. Dark place, 25 ℃, humidity 50 ~
When the culture medium was cultivated for 30 days under the condition of 60%, the mycelium spread over the entire bottle. Further, the culture was continued under the same conditions for 37 days to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, Culturing was continued for 9 days under the condition of 20 lux to obtain fruit body primordia.
Cultivation was continued for 3 days to obtain fruiting bodies. The yield of the fruiting body obtained was 157 g, and no bitterness was felt in the sensory test of the fruiting body.
【0044】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3
0g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。Example 4 100 ml of liquid PGY medium was inoculated with lyophilum ulmarium K-0429 (FERM P-12570) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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%, hyphae spread to the entire bottle. Furthermore, culture was continued for 43 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water is discarded, and the conditions are 15 ° C., humidity 90 to 95%, and illuminance 20 Lux for 9 days. Continue culturing to obtain fruit body primordia,
The illuminance was further raised to 200 lux and the culture was continued for 13 days to obtain fruiting bodies. The yield of the fruiting body obtained was 184 g,
No bitterness was felt in the sensory test of fruiting bodies.
【0045】実施例5
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 5 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
, 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 Riofilum ulmarium K-0429. Dark place, 25
When the culture medium was cultured for 28 days under the condition of the temperature of 50 ° C. and the humidity of 50 to 60%, the mycelia were spread over the entire bottle. Further, the culture was continued under the same conditions for 37 days to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 9 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain fruit body primordia, and the illuminance was further raised to 200 lux for 13 days of culturing to obtain fruit bodies. The yield of the fruiting body obtained was 157 g, and no bitterness was felt in the sensory test of the fruiting body.
【0046】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−042
9の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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1
3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。Example 6 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-042.
9 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 28 days under the condition of 0%, the mycelium spread over the entire bottle. Further, the culture was continued under the same conditions for 37 days to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, Culturing was continued for 9 days under the condition of 20 lux to obtain fruit body primordia.
Cultivation was continued for 3 days to obtain fruiting bodies. The yield of the fruiting body thus obtained was 164 g, and no bitterness was felt in the sensory test of the fruiting body.
【0047】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3
0g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。Example 7 100 ml of a 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,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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 the dark at 25 ° C. and a humidity of 50 to 60%, hyphae were spread over the entire bottle. Further, the culture was continued for 35 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water is discarded, and the conditions are 15 ° C, humidity 90 to 95%, and illuminance 20 lux for 10 days. The culturing was continued to obtain a fruiting body primordium, the illuminance was further raised to 200 lux, and the cultivation was continued for 14 days to obtain a fruiting body. The yield of fruiting body obtained is 188 g.
No bitterness was felt in the sensory test of fruiting bodies.
【0048】実施例8
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 8 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
, 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 Riofilum ulmarium K-0207. Dark place, 25
When the culture medium was cultivated for 34 days at a temperature of 50 ° C. and a humidity of 50 to 60%, hyphae spread over the entire bottle. Further, the culture was continued for 31 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 9 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain a fruiting body primordium. Further, the illuminance was increased to 200 lux and culturing was continued for 14 days to obtain fruiting bodies. The yield of the fruiting body thus obtained was 161 g, and no bitterness was felt in the sensory test of the fruiting body.
【0049】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−020
7の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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。Example 9 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-020.
7 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultured for 33 days under the condition of 0%, the mycelium spread over the entire bottle. Further, the culture was continued for 32 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, The culturing was continued for 11 days under the condition of 20 lux to obtain fruit body primordia, and the illuminance was further raised to 200 lux for 13 days to continue cultivation to obtain fruit bodies. The yield of the fruiting body thus obtained was 178 g, and no bitterness was felt in the sensory test of the fruiting body.
【0050】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3
0g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。Example 10 100 ml of a liquid PGY medium was inoculated with riophyllum ulmarium K-0202 (FERM P-12980) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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 a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia were spread over the entire bottle. Further, the culture was continued for 35 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to supply water sufficiently, the excess tap water was discarded, and the conditions were 15 ° C., humidity 90 to 95%, and illuminance 20 Lux for 8 days. Continue culturing to obtain fruit body primordia,
The illuminance was further raised to 200 lux and the culture was continued for 13 days to obtain fruiting bodies. The yield of the fruiting body obtained was 187 g,
No bitterness was felt in the sensory test of fruiting bodies.
【0051】実施例11
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 11 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium K-0202. Dark place, 25
When the culture medium was cultivated for 40 days at a temperature of 50 ° C. and a humidity of 50% to 60%, hyphae were spread over the entire bottle. Further, the culture was continued for 40 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued under the conditions of humidity of 90 to 95% and illumination of 20 lux for 8 days to obtain fruit body primordium, and the illuminance was further raised to 200 lux to continue culture for 12 days to obtain fruit bodies. The yield of the fruiting body thus obtained was 154 g, and no bitterness was felt in the sensory test of the fruiting body.
【0052】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−020
2の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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1
3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。Example 12 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-020.
Two solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 36 days under the condition of 0%, the mycelium spread over the entire bottle. Furthermore, the culture was continued for 44 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, The illuminance was 20 lux, and the culture was continued for 8 days to obtain fruit body primordia.
Cultivation was continued for 3 days to obtain fruiting bodies. The yield of the fruiting body obtained was 162 g, and no bitterness was felt in the sensory test of the fruiting body.
【0053】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3
0g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。Example 13 100 ml of the liquid PGY medium was inoculated with lyophilum ulmarium K-0259 (FERM P-12981) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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 a dark place at 25 ° C. and a humidity of 50 to 60%, mycelia were spread over the entire bottle. Further, the culture was continued for 35 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to supply water sufficiently, the excess tap water was discarded, and the conditions were 15 ° C., humidity 90 to 95%, and illuminance 20 Lux for 8 days. Continue culturing to obtain fruit body primordia,
The illuminance was further raised to 200 lux and the culture was continued for 12 days to obtain fruiting bodies. The yield of the fruiting body obtained was 192 g,
No bitterness was felt in the sensory test of fruiting bodies.
【0054】実施例14
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 14 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium K-0259. Dark place, 25
When the culture medium was cultivated for 35 days at 50 ° C. and a humidity of 50 to 60%, mycelia spread to the entire bottle. Further, the culture was continued for 45 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 8 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain fruit body primordium. Further, the illuminance was increased to 200 lux and culturing was continued for 13 days to obtain fruit body. The yield of the fruiting body obtained was 162 g, and no bitterness was felt in the sensory test of the fruiting body.
【0055】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−025
9の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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1
3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。Example 15 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-025.
9 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 37 days under the condition of 0%, the mycelium spread over the entire bottle. Furthermore, culture was continued for 43 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, The illuminance was 20 lux, and the culture was continued for 8 days to obtain fruit body primordia.
Cultivation was continued for 3 days to obtain fruiting bodies. The yield of the fruiting body thus obtained was 165 g, and no bitterness was felt in the sensory test of the fruiting body.
【0056】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3
0g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。Example 16 100 ml of liquid PGY medium was inoculated with riophyllum ulmarium K-1004 (FERM P-12982) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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 the dark at 25 ° C. and a humidity of 50 to 60%, the mycelium spread to the entire bottle. Furthermore, the culture was continued for 50 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group, adding 20 ml of tap water and supplying water sufficiently, the excess tap water was discarded, and the conditions were 15 ° C., humidity 90 to 95%, and illuminance 20 lux for 11 days. The culturing was continued to obtain a fruiting body primordium, the illuminance was further raised to 200 lux, and the cultivation was continued for 15 days to obtain a fruiting body. The yield of fruiting body obtained is 188 g.
No bitterness was felt in the sensory test of fruiting bodies.
【0057】実施例17
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 17 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium K-1004. Dark place, 25
When the culture medium was cultivated for 32 days at a temperature of 50 ° C. and a humidity of 50 to 60%, hyphae were spread over the entire bottle. Further, the culture was continued for 48 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 10 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain fruit body primordium. Further, the illuminance was increased to 200 lux and culturing was continued for 15 days to obtain fruit body. The yield of the fruiting body thus obtained was 158 g, and no bitterness was felt in the sensory test of the fruiting body.
【0058】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−100
4の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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。Example 18 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then lyophilum ulmarium K-100.
4 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 28 days under the condition of 0%, the mycelium spread over the entire bottle. Further, the culture was continued for 52 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, And the illuminance was 20 lux, the culture was continued for 11 days to obtain a fruit body primordium, and the illuminance was further raised to 200 lux to continue the culture for 16 days to obtain a fruit body. The yield of the fruiting body thus obtained was 161 g, and no bitterness was felt in the sensory test of the fruiting body.
【0059】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3
0g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。Example 19 100 ml of liquid PGY medium was inoculated with riophyllum ulmarium K-1257 (FERM P-12983) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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 the dark at 25 ° C. and a humidity of 50 to 60%, the mycelium spread to the entire bottle. Furthermore, the culture was continued for 50 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group, adding 20 ml of tap water and supplying water sufficiently, the excess tap water was discarded, and the conditions were 15 ° C., humidity 90 to 95%, and illuminance 20 lux for 11 days. The culturing was continued to obtain a fruiting body primordium, the illuminance was further raised to 200 lux, and the cultivation was continued for 15 days to obtain a fruiting body. The yield of the fruiting body obtained is 186 g.
No bitterness was felt in the sensory test of fruiting bodies.
【0060】実施例20
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 20 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, the culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and inoculated with a solid inoculum of Riophyllum ulmarium K-1257. Dark place, 25
When the culture medium was cultivated for 32 days at a temperature of 50 ° C. and a humidity of 50 to 60%, hyphae were spread over the entire bottle. Further, the culture was continued for 48 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued under the conditions of humidity of 90 to 95% and illumination of 20 lux for 9 days to obtain a fruiting body primordium, and the illuminance was further raised to 200 lux to continue culturing for 16 days to obtain a fruiting body. The yield of the fruiting body thus obtained was 160 g, and no bitterness was felt in the sensory test of the fruiting body.
【0061】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−125
7の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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。Example 21 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-125.
7 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 28 days under the condition of 0%, the mycelium spread over the entire bottle. Further, the culture was continued for 52 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, And the illuminance was 20 lux, the culture was continued for 10 days to obtain a fruit body primordium, and the illuminance was further increased to 200 lux to continue the culture for 14 days to obtain a fruit body. The yield of the fruiting body obtained was 157 g, and no bitterness was felt in the sensory test of the fruiting body.
【0062】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3
0g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。Example 22 100 ml of liquid PGY medium was inoculated with riophyllum ulmarium K-6804 (FERM P-12984) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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 the dark at 25 ° C. and a humidity of 50 to 60%, the mycelium spread to the entire bottle. Furthermore, the culture was continued for 30 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water is discarded, and the conditions are 15 ° C, humidity 90 to 95%, and illuminance 20 lux for 10 days. The culturing was continued to obtain a fruiting body primordium, the illuminance was further raised to 200 lux, and the cultivation was continued for 16 days to obtain a fruiting body. The yield of the fruiting body obtained is 195 g.
No bitterness was felt in the sensory test of fruiting bodies.
【0063】実施例23
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 23 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium K-6804. Dark place, 25
When the culture medium was cultured for 28 days under the condition of the temperature of 50 ° C. and the humidity of 50 to 60%, the mycelia were spread over the entire bottle. Further, the culture was continued for 32 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 10 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain fruit body primordium. Further, the illuminance was increased to 200 lux and culturing was continued for 15 days to obtain fruit body. The yield of the fruiting body thus obtained was 163 g, and no bitterness was felt in the sensory test of the fruiting body.
【0064】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−680
4の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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1
6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。Example 24 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-680.
4 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 28 days under the condition of 0%, the mycelium spread over the entire bottle. Further, the culture was continued for 32 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, Culturing was continued for 9 days under the condition of 20 lux to obtain fruit body primordia.
Cultivation was continued for 6 days to obtain fruiting bodies. The yield of the fruiting body thus obtained was 171 g, and no bitterness was felt in the sensory test of the fruiting body.
【0065】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3
0g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。Example 25 100 ml of a liquid PGY medium was inoculated with lyophilum ulmarium K-6806 (FERM P-12985) and cultured at 25 ° C. for 10 days to obtain a liquid inoculum. on the other hand,
Corncob meal 130g, Mamekawa 60g, Bran 3
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, the culture medium was sterilized by high pressure steam 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%, hyphae spread to the entire bottle. Further, the culture was continued for 38 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water is discarded, and the conditions are 15 ° C., humidity 90 to 95%, and illuminance 20 Lux for 9 days. Continue culturing to obtain fruit body primordia,
The illuminance was further raised to 200 lux and the culture was continued for 15 days to obtain fruiting bodies. The yield of the fruiting body obtained was 191 g,
No bitterness was felt in the sensory test of fruiting bodies.
【0066】実施例26
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Example 26 50 g of softwood sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium K-6806. Dark place, 25
When the culture medium was cultured for 30 days at a temperature of 50 ° C. and a humidity of 50% to 60%, the mycelium spread to the entire bottle. Further, the culture was continued for 40 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 9 days under the conditions of humidity of 90 to 95% and illumination of 20 lux to obtain fruit body primordium, and the illuminance was further raised to 200 lux to continue culturing for 15 days to obtain fruiting bodies. The yield of the fruiting body obtained was 162 g, and no bitterness was felt in the sensory test of the fruiting body.
【0067】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−680
6の固体種菌を接種した。暗所、25℃、湿度50〜6
0%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1
4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。Example 27 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C. for 60 minutes, cooled to room temperature, and then Lyophilum ulmarium K-680.
6 solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultured for 31 days under the condition of 0%, the mycelium spread to the entire bottle. Further, the culture was continued for 39 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, Culturing was continued for 9 days under the condition of 20 lux to obtain fruit body primordia.
Cultivation was continued for 4 days to obtain fruiting bodies. The yield of the fruiting body thus obtained was 165 g, and no bitterness was felt in the sensory test of the fruiting body.
【0068】対照例1
コーンコブミール130g、マメカワ60g、フスマ3
0g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。Control Example 1 130 g of corn cob meal, 60 g of mamekawa, 3 of fusuma
0 g was mixed well and the water content was adjusted to 62% with tap water. It was pressed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the mouth of the bottle.
After puncturing, 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 Riofilum ulmarium Lu 1-2. Dark place, 25
When the culture medium was cultivated for 35 days at 50 ° C. and a humidity of 50 to 60%, mycelia spread to the entire bottle. Furthermore, the culture was continued for 50 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 9 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain fruit body primordia, and the illuminance was further raised to 200 lux for 13 days of culturing to obtain fruit bodies. The yield of the fruiting body thus obtained was 178 g, and bitterness was felt in the sensory test of the fruiting body.
【0069】対照例2
針葉樹オガクズ50g、綿実殻粉砕物50g、米糠10
0g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。Control Example 2 50 g of coniferous sawdust, 50 g of crushed cottonseed shell, 10 rice bran
0 g was mixed well and the water content was adjusted to 63% with tap water. It was packed into a polypropylene 850 ml wide mouth bottle, and the diameter was 1 cm downward from the center of the bottle mouth.
After puncturing, 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 Riofilum ulmarium Lu 1-2. Dark place, 25
When the culture medium was cultured for 30 days at a temperature of 50 ° C. and a humidity of 50% to 60%, the mycelium spread to the entire bottle. Further, cultivation was continued for 55 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating base and adding 20 ml of tap water to supply water sufficiently, the surplus tap water was discarded and the temperature was raised to 15 ° C.
Cultivation was continued for 9 days under conditions of humidity of 90 to 95% and illuminance of 20 lux to obtain a fruiting body primordium. Further, the illuminance was increased to 200 lux and culturing was continued for 14 days to obtain fruiting bodies. The yield of the fruiting body obtained was 140 g, and bitterness was felt in the sensory test of the fruiting body.
【0070】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50〜6
0%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。Comparative Example 3 100 g of coniferous sawdust and 70 g of Mamekawa were mixed well and the water content was adjusted to 63% with tap water.
After compressing it into a polypropylene 850 ml wide-mouthed bottle, punch a hole with a diameter of 1 cm downward from the center of the bottle mouth,
The culture medium was sterilized by high-pressure steam at 120 ° C for 60 minutes, cooled to room temperature, and then lyophilum ulmarium Lu 1-
Two solid inoculums were inoculated. Dark place, 25 ° C, humidity 50-6
When the culture medium was cultivated for 30 days under the condition of 0%, the mycelium spread over the entire bottle. Further, cultivation was continued for 55 days under the same conditions to obtain a fruiting body-generating group. After removing 1 cm of the upper hypha layer of the fruiting body-generating group and adding 20 ml of tap water to sufficiently supply water, surplus tap water was discarded, and the temperature was 15 ° C. and the humidity was 90 to 95.
%, And the illuminance was 20 lux, the culture was continued for 10 days to obtain fruit body primordia, and the illuminance was further increased to 200 lux to continue culture for 13 days to obtain fruit bodies. The yield of the fruiting body thus obtained was 153 g, and bitterness was felt in the sensory test of the fruiting body.
【0071】[0071]
【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。As described above, according to the present invention, it is possible to obtain Riofilum ulmarium fruiting bodies with reduced bitterness of fruiting bodies even when cultivated using a high bittering medium which could not be used conventionally. It is possible to provide lyophilum ulmarium fruiting bodies and mycelium of high quality with high yield industrially.
───────────────────────────────────────────────────── フロントページの続き 微生物の受託番号 FERM P−12985 前置審査 (72)発明者 木野 真奈美 滋賀県大津市瀬田3丁目4番1号 寳酒 造株式会社中央研究所内 (72)発明者 松井 侑 滋賀県大津市瀬田3丁目4番1号 寳酒 造株式会社中央研究所内 (72)発明者 森田 日出男 滋賀県大津市瀬田3丁目4番1号 寳酒 造株式会社中央研究所内 (58)調査した分野(Int.Cl.7,DB名) C12N 1/14 A01G 1/04 A01H 15/00 ─────────────────────────────────────────────────── ─── Continuation of front page microbial consignment number FERM P-12985 Preliminary examination (72) Inventor Manami Kino 3-4-1 Seta 3-chome Seta, Otsu City, Shiga Prefecture In-house Research Laboratories (72) Inventor Matsui YU, Central 3-chome, Seta 3-chome, Otsu City, Shiga Prefecture Inventor, Hideo Morita (72) Inventor Hideo Morita 3-chome 4-chome Seta, Otsu City, Shiga Prefecture (58) Survey Areas (Int.Cl. 7 , DB name) C12N 1/14 A01G 1/04 A01H 15/00
Claims (2)
の苦味が、少なくとも一種の高苦味化培地について、
0.000008M 硫酸キニーネよりも苦くなく、か
つ下記の菌株、又はこれらの変異株から選択されるリオ
フィラム ウルマリウム新菌株を培地に接種し、菌糸体
を生成させることを特徴とするリオフィラム ウルマリ
ウム新菌株の培養方法。 リオフィラム ウルマリウム K−0429(FERM
P−12570)、 リオフィラム ウルマリウム K−0207(FERM
P−12569)、 リオフィラム ウルマリウム K−0202(FERM
P−12980)、 リオフィラム ウルマリウム K−0259(FERM
P−12981)、 リオフィラム ウルマリウム K−1004(FERM
P−12982)、 リオフィラム ウルマリウム K−1257(FERM
P−12983)、 リオフィラム ウルマリウム K−6804(FERM
P−12984)、 リオフィラム ウルマリウム K−6806(FERM
P−12985)1. The bitterness of fruiting bodies obtained by culturing in a high bittering medium is at least one high bittering medium,
Culture of a new strain of Riofilum ulmarium, which is less bitter than 0.000008M quinine sulfate and inoculates a medium with a new strain of Riofilum ulmarium selected from the following strains or mutants thereof to produce mycelia: Method. Riofilum Ulmarium K-0429 (FERM
P-12570), Riofilum ulmarium K-0207 (FERM
P-12569), Riofilum ulmarium K-0202 (FERM
P-12980), Riofilum ulmarium K-0259 (FERM
P-12981), Riofilum ulmarium K-1004 (FERM
P-12982), Riofilum ulmarium K-1257 (FERM
P-12983), Riofilum ulmarium K-6804 (FERM
P-12984), Riofilum ulmarium K-6806 (FERM
P-12985)
の苦味が、少なくとも一種の高苦味化培地について、
0.000008M 硫酸キニーネよりも苦くなく、か
つ下記の菌株、又はこれらの変異株から選択されるリオ
フィラム ウルマリウム新菌株を培地に接種し、子実体
を生成させることを特徴とするリオフィラム ウルマリ
ウム新菌株の子実体の栽培方法。 リオフィラム ウルマリウム K−0429(FERM
P−12570)、 リオフィラム ウルマリウム K−0207(FERM
P−12569)、 リオフィラム ウルマリウム K−0202(FERM
P−12980)、 リオフィラム ウルマリウム K−0259(FERM
P−12981)、 リオフィラム ウルマリウム K−1004(FERM
P−12982)、 リオフィラム ウルマリウム K−1257(FERM
P−12983)、 リオフィラム ウルマリウム K−6804(FERM
P−12984)、 リオフィラム ウルマリウム K−6806(FERM
P−12985)2. The bitterness of fruiting bodies obtained by culturing in a high bittering medium is at least one high bittering medium,
A child of a new strain of Riofilum ulmarium, which is less bitter than 0.000008M quinine sulfate and inoculates a medium with a new strain of Riofilum ulmarium selected from the following strains or mutants thereof to produce fruiting bodies. Cultivation method of substance. Riofilum Ulmarium K-0429 (FERM
P-12570), Riofilum ulmarium K-0207 (FERM
P-12569), Riofilum ulmarium K-0202 (FERM
P-12980), Riofilum ulmarium K-0259 (FERM
P-12981), Riofilum ulmarium K-1004 (FERM
P-12982), Riofilum ulmarium K-1257 (FERM
P-12983), Riofilum ulmarium K-6804 (FERM
P-12984), Riofilum ulmarium K-6806 (FERM
P-12985)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP23163492A JP3436768B2 (en) | 1991-11-19 | 1992-08-07 | Culture and cultivation method of new strain |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3-329878 | 1991-11-19 | ||
JP32987891 | 1991-11-19 | ||
JP23163492A JP3436768B2 (en) | 1991-11-19 | 1992-08-07 | Culture and cultivation method of new strain |
Related Child Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP09747099A Division JP3229287B2 (en) | 1991-11-19 | 1999-04-05 | Mushroom artificial cultivation method |
JP2002012273A Division JP3571697B2 (en) | 1991-11-19 | 2002-01-22 | New strain, culture and cultivation method |
JP2002334490A Division JP3571710B2 (en) | 1991-11-19 | 2002-11-19 | Culture and cultivation method of new strain |
JP2003126647A Division JP3539953B2 (en) | 1991-11-19 | 2003-05-01 | New strain of lyophilum ulmarium |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH05268942A JPH05268942A (en) | 1993-10-19 |
JP3436768B2 true JP3436768B2 (en) | 2003-08-18 |
Family
ID=27806816
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP23163492A Expired - Lifetime JP3436768B2 (en) | 1991-11-19 | 1992-08-07 | Culture and cultivation method of new strain |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP3436768B2 (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101044345B1 (en) * | 2002-05-10 | 2011-06-29 | 다카라 바이오 가부시키가이샤 | Artificial cultivation of mushroom |
JP4842235B2 (en) * | 2002-05-10 | 2011-12-21 | タカラバイオ株式会社 | Mushroom artificial cultivation method |
KR20080105001A (en) | 2007-05-29 | 2008-12-03 | 다카라 바이오 가부시키가이샤 | Mushroom bed cultivation of mushroom |
KR20110109959A (en) * | 2010-03-31 | 2011-10-06 | 다카라 바이오 가부시키가이샤 | Hypsizigus marmoreus strain and method for manufacturing of fruiting body of hypsizigus marmoreus |
-
1992
- 1992-08-07 JP JP23163492A patent/JP3436768B2/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
JPH05268942A (en) | 1993-10-19 |
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