JP2011152150A - Method for artificially cultivating mushroom - Google Patents
Method for artificially cultivating mushroom Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/10—Mycorrhiza; Mycorrhizal associations
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
Abstract
Description
本発明は、きのこの人工栽培方法に関し、更に詳しくは収量及び品質の優れたものを得ることができる、きのこの人工栽培用培養基及び該培養基を用いたきのこの人工栽培方法、並びに良好な子実体形成能を有するきのこ菌株及び該菌株を用いるきのこの人工栽培方法に関する。 TECHNICAL FIELD The present invention relates to a method for artificial cultivation of mushrooms, and more specifically, a culture medium for artificial cultivation of mushrooms, a method for artificial cultivation of mushrooms using the culture medium, and a good fruiting body capable of obtaining a product with excellent yield and quality. The present invention relates to a mushroom strain having a forming ability and an artificial cultivation method of a mushroom using the strain.
近年、シイタケ、エノキタケ、ヒラタケ、ナメコ、マイタケ、リオフィラム ウルマリウム、ハタケシメジ等の食用きのこは、人工栽培方法が発達してきている。人工栽培方法は大きく分類して原木人工栽培方法と菌床人工栽培方法があり、前者はコナラ、ブナ、クヌギ等の原木をホダ木として使用する栽培方法であり、後者はオガクズ、コーンコブ等を培地間隙の保持と保水のための培地支持体用基材とし、これに米糠、小麦フスマ、コーンブラン等の栄養源としての基材を混合して調製した培地をビン、袋、箱などの容器に充填した固形培養基を用いる栽培方法である。
菌床人工栽培方法において、これらの基材を使用した培地では、安価な材料を選択すると十分な収量が得られないことが多い。例えば、最も安価な基材として、オガクズと米糠のみを使用し、リオフィラム ウルマリウムを８５０ｍｌビンで人工栽培した場合の収量は７０ｇ前後であり、安価ではあるが、満足できるものではなかった。
In recent years, edible mushrooms such as shiitake mushrooms, enokitake mushrooms, oyster mushrooms, sea cucumbers, maitake mushrooms, lyophilum urumarium, and bamboo shimeji mushrooms have developed artificial cultivation methods. Artificial cultivation methods can be broadly classified into raw wood artificial cultivation methods and fungus bed artificial cultivation methods, the former is a cultivation method that uses raw wood such as konara, beech and kunugi as hoda trees, and the latter is a medium that uses sawdust, corn cob, etc. Use as a base material for a medium support for maintaining gaps and retaining water, and mix the medium as a nutrient source such as rice bran, wheat bran, and corn bran into a container such as a bottle, bag, or box. It is a cultivation method using a filled solid culture medium.
In a method for artificial cultivation of fungus beds, in a medium using these base materials, a sufficient yield is often not obtained when an inexpensive material is selected. For example, as the cheapest base material, only sawdust and rice bran were used, and when liophyllum urumarium was artificially grown in an 850 ml bottle, the yield was around 70 g, which was inexpensive but was not satisfactory.
本発明の目的は、きのこの人工栽培において従来の方法よりも安価な材料を使用して、多くの収量を得ることのできるきのこの培養基及び該培養基を使用した人工栽培方法を提供し、特に該培養基で良好な子実体形成能を有する菌株を提供することにある。 An object of the present invention is to provide a mushroom culture medium capable of obtaining a large yield by using a material cheaper than the conventional method in artificial cultivation of mushrooms, and an artificial cultivation method using the culture medium. It is an object of the present invention to provide a strain having a good fruiting ability in a culture medium.
本発明を概説すれば、本発明は以下の各発明を包含する。
（１）１種類以上のセリ科植物由来の処理物からなる基材を含有することを特徴とするきのこの人工栽培用培養基。
（２）上記（１）項に記載のきのこの人工栽培用培養基を使用することを特徴とするきのこの人工栽培方法。
（３）１種類以上のセリ科植物由来の処理物からなる基材を含有する培養基で人工栽培したときに、良好な子実体形成能を有することを特徴とするリオフィラム ウルマリウム菌株。
（４）Ｌｕ１−１７２（ＦＥＲＭ ＢＰ−８３５４）、Ｌｕ１−１７３（ＦＥＲＭ ＢＰ−８３５５）、Ｌｕ１−１７４（ＦＥＲＭ ＢＰ−８３５６）、Ｌｕ１−１８１（ＦＥＲＭ ＢＰ−８３５７）、又はこれらの変異株から選択される菌株であることを特徴とするリオフィラム ウルマリウム新菌株。
（５）上記（３）又は（４）項に記載の菌株を用いることを特徴とするきのこの人工栽培方法。
（６）人工栽培方法が、セリ科植物由来の処理物からなる基材を含有する培養基を用いる方法であることを特徴とする（５）項に記載のきのこの人工栽培方法。
（７）上記（１）項に記載のきのこの人工栽培用培養基を使用してきのこを栽培する工程を包含することを特徴とするきのこの製造方法。
（８）きのこが（３）又は（４）項に記載されるリオフィラム ウルマリウム菌株である、（７）項に記載の製造方法。
本明細書において、基材とはきのこの人工栽培用培養基を構成する要素の中で水以外の構成要素のことをいい、例えば培地支持体や栄養源のことをいう。
If the present invention is outlined, the present invention includes the following inventions.
(1) A culture medium for artificial cultivation of mushrooms, comprising a base material made of a processed product derived from one or more kinds of cereals.
(2) A method for artificial cultivation of a mushroom characterized by using the culture medium for artificial cultivation of a mushroom described in (1) above.
(3) A liophyllum ulmarium strain characterized by having a good fruiting body formation ability when artificially cultivated in a culture medium containing a base material made of a processed product derived from one or more kinds of cereals.
(4) Selected from Lu1-172 (FERM BP-8354), Lu1-173 (FERM BP-8355), Lu1-174 (FERM BP-8356), Lu1-181 (FERM BP-8357), or mutants thereof A new strain of lyophilum urmarium, characterized in that
(5) A method for artificial cultivation of mushrooms characterized by using the strain described in (3) or (4) above.
(6) The artificial cultivation method for mushrooms according to item (5), wherein the artificial cultivation method is a method using a culture medium containing a base material made of a processed product derived from a ciraceae plant.
(7) A method for producing mushrooms, comprising a step of cultivating mushrooms using the culture medium for artificial cultivation of mushrooms according to item (1).
(8) The production method according to (7), wherein the mushroom is the liophyllum ulmarium strain described in (3) or (4).
In the present specification, the base material refers to a constituent element other than water among the constituent elements of the culture medium for artificial cultivation of mushrooms, for example, a medium support or a nutrient source.
本発明者らは、前記の目的を達成するため、種々の材料を基材として使用した結果、安価で多くの収量を得ることのできる基材源として、セリ科植物を見出した。更に各種条件で鋭意検討した結果、セリ科植物由来の植物を基材として単独で使用するより、２種類以上の基材から構成されるきのこの人工栽培用培養基において、１種類以上のセリ科植物由来の処理物からなる基材と、当該セリ科植物由来の処理物からなる基材以外の１種類以上の基材とを使用することによってより多くの収量が得られることを見出した。また、当該培養基を用いたきのこの人工栽培では栽培日数が短く、機能性成分に富むきのこを得ることができる。
更に、本発明者らは交配育種を行い、得られた交配株の中からセリ科植物由来の処理物からなる基材を含有する培養基で人工栽培したときに、特に良好な子実体形成能を有するリオフィラム ウルマリウム新菌株を見出し、本発明を完成させた。
As a result of using various materials as a base material in order to achieve the above-mentioned object, the present inventors have found a celery family plant as a base material source capable of obtaining a large yield at a low cost. Furthermore, as a result of diligent examinations under various conditions, one or more kinds of celery family plants are used in the culture medium for artificial cultivation of mushrooms composed of two or more kinds of base materials, rather than using a plant derived from the family of cereals alone as a base material. It has been found that a higher yield can be obtained by using a base material composed of a processed product derived from the origin and one or more types of base materials other than the base material composed of the processed product derived from the family Aceraceae. In addition, mushroom artificial cultivation using the culture medium has short cultivation days, and mushrooms rich in functional components can be obtained.
Furthermore, the present inventors have carried out cross breeding, and when artificially cultivated with a culture medium containing a base material made of a processed product derived from a family of Apiaceae from the obtained cross strains, particularly good fruiting body forming ability is obtained. The present invention was completed by finding a new strain of liophyllum ulmarium.
本発明により、きのこの人工栽培において、本発明のきのこの人工栽培用培養基を使用することによって、より多くの収量のきのこを得ることが可能になった。
また、セリ科植物由来の処理物からなる基材を含有する培養基で良好な子実体形成能を有するリオフィラム ウルマリウム新菌株が提供され、高品質のリオフィラム ウルマリウムを得ることが可能になった。
According to the present invention, in the artificial cultivation of mushrooms, it becomes possible to obtain a higher yield of mushrooms by using the culture medium for artificial cultivation of the mushrooms of the present invention.
In addition, a new lyophilum ulmarium strain having a good fruiting body forming ability with a culture medium containing a base material made of a processed product derived from a ciraceae plant has been provided, and it has become possible to obtain high-quality lyophilum ulmarium.
以下、本発明を具体的に説明する。
本発明のきのこの人工栽培用培養基は２種類以上の基材から構成され、１種類以上のセリ科植物由来の処理物からなる基材と、当該セリ科植物由来の処理物からなる基材以外の１種類以上の基材とから構成される。本発明で使用されるセリ科植物とは被子植物類セリ目セリ科に属する植物であり、セリ、ミツバ、アシタバ、シシウド、ニンジン、セロリ等が例示される。これらの植物としては、その植物体全体を用いることができ、またその一部として、特に限定はないが、葉、茎、根又は根茎を使用することもできる。更に、本発明で用いることができるセリ科植物は新鮮物でも乾燥物でもよい。
本発明において、セリ科植物由来の処理物としては、細断、成形、加工、圧搾、搾汁等により得られる上記セリ科植物の処理物が例示され、これらの処理により分別された一部分を使用してもよい。また、これらの処理により得られるそれぞれの処理物を混合して使用しても良い。その処理物としては入手可能なあらゆる形状のものが使用できる。この中でも、成形時に生じる成形屑、加工時に生じる加工屑、搾汁時に生じる搾汁粕等は、通常は廃棄されるものであるため通常安価で入手でき、培養基のコストを低く抑えられることから、これら成形屑、加工屑、搾汁粕等を使用することが好ましい。
また、本発明に使用されるセリ科植物由来の処理物としては、特に限定はないが、栽培されるきのこの生育を妨げないものが特に好適に使用される。
Hereinafter, the present invention will be specifically described.
The culture medium for artificial cultivation of the mushroom of the present invention is composed of two or more kinds of base materials, other than the base material made of one or more kinds of processed products derived from the aceae plant and the base material made of the processed products derived from the celery family. And one or more kinds of base materials. The Aceraceae plant used in the present invention is a plant belonging to the Angiospermaceae Aceraceae, and examples include Acer, Mitsuba, Ashitaba, Shishiudo, Carrot, Celery and the like. As these plants, the whole plant can be used, and as a part thereof, there is no particular limitation, but leaves, stems, roots or rhizomes can also be used. Furthermore, the Apiaceae plants that can be used in the present invention may be fresh or dried.
In the present invention, as a processed product derived from the celery family plant, the processed product of the above celery family plant obtained by shredding, molding, processing, pressing, squeezing, etc. is exemplified, and a part separated by these treatments is used. May be. Moreover, you may use it, mixing each processed material obtained by these processes. The processed product can be any shape available. Among these, the molding waste generated at the time of molding, the processing waste generated at the time of processing, the juice lees generated at the time of squeezing, etc. are usually discarded, so they can usually be obtained at a low price, and the cost of the culture medium can be kept low. It is preferable to use these molding wastes, processing wastes, squeezed rice cakes and the like.
Moreover, there is no limitation in particular as a processed product derived from the Apiaceae plant used for this invention, The thing which does not prevent the growth of the mushroom cultivated is used especially suitably.
本発明において、セリ科植物としては、特に好適にはアシタバが使用できる。
アシタバはセリ科の大型多年生草本であり、その根、茎、葉に多種多様のカルコン類やクマリン類等の機能性成分を含むことが知られている。アシタバのもつ生理作用としては、高血圧予防作用、抗潰瘍作用、胃酸分泌抑制作用、抗癌作用、癌予防効果等が知られており、更に上記のカルコン類は神経栄養因子産生増強作用を示すことが知られており、アルツハイマー症や痴呆症等の神経性疾患の治療又は予防への効果が期待されている（国際公開第０１／７６６１４号パンフレット）。本発明においては、このような機能性食品素材であるアシタバをきのこの人工栽培用培養基材として使用することにより、これらの機能性成分を含有するきのこを得ることができる。
In the present invention, Ashitaba can be particularly preferably used as the Apiaceae plant.
Ashitaba is a large perennial herb belonging to the Apiaceae family, and it is known that its roots, stems and leaves contain various functional components such as chalcones and coumarins. Known physiological effects of Ashitaba include antihypertensive action, anti-ulcer action, gastric acid secretion inhibitory action, anticancer action, cancer prevention effect, etc. Further, the above chalcones show neurotrophic factor production enhancing action And is expected to have an effect on the treatment or prevention of neurological diseases such as Alzheimer's disease and dementia (WO01 / 76614). In the present invention, mushrooms containing these functional ingredients can be obtained by using Ashitaba, which is such a functional food material, as a culture substrate for artificial cultivation of mushrooms.
本発明において、セリ科植物由来の処理物からなる基材以外の基材としては、上記のセリ科植物全体若しくはその一部から由来の処理物以外のものであれば特に限定はなく、通常のきのこの人工栽培に用いられる基材であれば好適に用いることができる。
この中でも吸水性の高い素材が特に好適であり、この例として、針葉樹オガクズ、広葉樹オガクズ、チップダスト、コーンコブ、綿実殻、バガス、アルファルファ、ビートパルプ、モミガラ、チモシー、オカラ、マメカワ、シリカゲル、多糖類ゲル、高吸水ポリマー、きのこ原木栽培後の廃ホダ木、きのこ菌床人工栽培後の廃菌床等を挙げることができる。また、吸水性の高い基材であれば上記以外の材料も好適である。
更に、本発明の培養基は、セリ科植物由来の処理物からなる基材以外の基材として、吸水性の良否にかかわらず、栄養源、ｐＨ調整剤、菌糸生長促進剤、増収剤等を添加してもよい。
これらはきのこの人工栽培に用いることができるものであれば特に限定はなく、栄養源の例としては米糠、小麦フスマ、コーンブラン、ホミニーフィード、バンサイ、オカラ、マメカワ等を挙げることができる。また、ｐＨ調整剤、菌糸成長促進剤、及び増収剤の一例として、それぞれ、炭酸カルシウム、クエン酸、及びメタケイ酸アルミン酸マグネシウムを挙げることができる。
本発明においては、これらのセリ科植物由来の処理物からなる基材以外の基材は１種のみを使用してもよいし、２種以上を混合して使用してもよい。また、高吸水性の基材とその他の基材を混合して使用してもよい。
In the present invention, the base material other than the base material composed of the processed product derived from the family Aceraceae is not particularly limited as long as it is other than the processed product derived from the whole Asteraceae plant or a part thereof. Any base material used for artificial cultivation of mushrooms can be suitably used.
Of these, materials with high water absorption are particularly suitable, and examples include coniferous sawdust, hardwood sawdust, chip dust, corn cob, cottonseed husk, bagasse, alfalfa, beet pulp, momigal, timothy, okara, mamekawa, silica gel, many Examples thereof include sugar gels, superabsorbent polymers, waste hodder trees after cultivation of mushroom raw wood, and waste fungus beds after artificial cultivation of mushroom fungus beds. In addition, materials other than those described above are also suitable as long as the substrate has high water absorption.
Furthermore, the culture medium of the present invention includes a nutrient source, a pH adjuster, a mycelial growth promoter, a yield enhancer, etc., as a substrate other than a substrate made of a processed product derived from a ciraceae plant, regardless of whether water absorption is good or bad. May be.
There are no particular limitations on these mushrooms as long as they can be used for artificial cultivation, and examples of nutrient sources include rice bran, wheat bran, corn bran, hominy feed, bansai, okara, mameka and the like. Moreover, calcium carbonate, a citric acid, and magnesium aluminate metasilicate can be mentioned as an example of a pH adjuster, a mycelium growth promoter, and a yield enhancer, respectively.
In the present invention, the base material other than the base material made of a processed product derived from these celery family plants may be used alone or in combination of two or more. Moreover, you may use it, mixing a highly water-absorbing base material and another base material.
本発明において、セリ科植物由来の処理物からなる基材と、セリ科植物由来の処理物からなる基材以外の基材との使用比率は、特に限定はないが、乾燥重量比でセリ科植物由来の処理物からなる基材１に対して、セリ科植物由来の処理物からなる基材以外の基材は望ましくは０．０５〜４０、更に望ましくは０．２〜２０がよい。 In the present invention, there is no particular limitation on the use ratio of the base material made of the processed product derived from the family Aceraceae and the base material other than the base material made of the processed product derived from the Asteraceae plant. The base material 1 made of a plant-derived processed product is preferably 0.05 to 40, more preferably 0.2 to 20 for a substrate other than the base material made of a processed product derived from a celery family plant.
本発明を実施するためのきのこの人工栽培法としては、袋、ビン、箱等を容器として用いる人工栽培方法が挙げられ、栽培の形態としては温度、湿度等を人工的に制御した空調施設栽培（周年栽培）、屋外の林地、畑地等に設置又は埋設する自然栽培等を挙げることができるが、本発明はこれらの人工栽培方法や栽培の形態に何ら制約されるものではない。一例としてビンを容器として用いる空調施設栽培を例として説明すると、培地調製、ビン詰め、殺菌、接種、培養、発生の各工程からなる。
培地調製とは１種類以上のセリ科植物由来の処理物からなる基材と、セリ科植物由来の処理物からなる基材以外の基材を混合してかくはんし、水分調整して培養基を製造する工程であり、培養基の水分含量は５０〜８５％、好ましくは６０〜７５％がよい。
ビン詰めとは培養基をビンに詰める工程であり、容量が２００〜１２００ｍｌ、好ましくは５００〜１０００ｍｌの耐熱性広ロビンに培養基を入れ、中央に穴を空け、打栓する工程をいう。ビンに入れる培養基の量はビンの容量、形状によって異なるが、例えばポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕を使用した場合、乾燥重量で１００〜３００ｇ、好ましくは１５０〜２５０ｇがよい。
殺菌とは培養基中の微生物を死滅させる行程であれば良く、通常常圧殺菌では９８℃で４〜１２時間、高圧殺菌では１０１〜１２５℃で３０〜９０分間行われる。
接種とは殺菌後放冷した培養基に固体種菌又は液体種菌５〜５０ｍｌを植え付ける工程である。
培養とはきのこの菌糸を生育、まん延、更に必要に応じて熟成後子実体発生基を得る工程であり、通常接種済みの培養基を１８〜２８℃、湿度４０〜８０％において行う。この工程はきのこの種類又は菌株により適宜選択すればよいが、通常２０〜１２０日間行われる。
発生とは子実体発生基を子実体形成に適当な条件に置いて子実体を発生させる工程であり、通常子実体形成を促進するために、菌掻き、加水、加湿、光照射等を行う。これらは常法に従って行えばよい。
以上、人工栽培方法についてビン栽培を例として詳細に述べたが、本発明は上記方法にのみ制約を受けるものではない。
Examples of the artificial cultivation method for mushrooms for carrying out the present invention include an artificial cultivation method using bags, bottles, boxes and the like as containers, and the cultivation mode is air conditioning facility cultivation in which temperature, humidity and the like are artificially controlled. (Anniversary cultivation), natural cultivation that is installed or buried in outdoor forest land, upland field, and the like can be mentioned, but the present invention is not limited to these artificial cultivation methods and cultivation forms. As an example, air-conditioning facility cultivation using a bottle as a container will be described as an example, which includes the steps of medium preparation, bottle filling, sterilization, inoculation, culture, and generation.
Medium preparation refers to the production of a culture medium by mixing a substrate made of a processed product derived from one or more aceae plants and a substrate other than a substrate made of a processed product derived from a celery family, and adjusting the water content. The water content of the culture medium is 50 to 85%, preferably 60 to 75%.
The bottling is a step of filling a culture medium into a bottle, which is a step of placing the culture medium in a heat-resistant wide robin having a capacity of 200 to 1200 ml, preferably 500 to 1000 ml, making a hole in the center, and plugging. The amount of culture medium put into the bottle varies depending on the capacity and shape of the bottle. For example, when a polypropylene 850 ml bottle (trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.) is used, the dry weight is 100 to 300 g. Preferably 150-250g is good.
Sterilization may be a process of killing microorganisms in the culture medium, and is usually performed at 98 ° C. for 4 to 12 hours for normal pressure sterilization and at 101 to 125 ° C. for 30 to 90 minutes for high pressure sterilization.
Inoculation is a process of planting 5 to 50 ml of solid inoculum or liquid inoculum on a culture medium that has been allowed to cool after sterilization.
Cultivation is a process of growing, spreading, and, if necessary, obtaining a fruit body generating group after maturation. Usually, the inoculated culture medium is carried out at 18 to 28 ° C. and humidity of 40 to 80%. This step may be appropriately selected depending on the type or strain of the mushroom, but is usually performed for 20 to 120 days.
The generation is a step of generating a fruiting body by placing the fruiting body generating group under conditions suitable for the fruiting body formation. Usually, in order to promote the fruiting body formation, bacterial scraping, water addition, humidification, light irradiation and the like are performed. These may be carried out according to ordinary methods.
As mentioned above, although the bottle cultivation was described in detail about the artificial cultivation method as an example, this invention does not receive a restriction | limiting only by the said method.
本発明におけるきのこの種類及び菌株については特に限定されるものではないが、きのこの種類としてはシイタケ、エノキタケ、ヒラタケ、ナメコ、マイタケ、リオフィラム ウルマリウム、ハタケシメジ等を挙げることができる。きのこの菌株としては市販の菌株でも、野生の子実体よりの組織分離株でも、選抜、交配、細胞融合、遺伝子組換え等の方法により育種した株でもよい。例えば、リオフィラム ウルマリウム菌株であるリオフィラム ウルマリウム Ｋ−０２５９、後述する新菌株であるリオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株を使用することができる。なお、リオフィラム ウルマリウム Ｋ−０２５９はＬｙｏｐｈｙｌｌｕｍ ｕｌｍａｒｉｕｍ Ｋ−０２５９と表示され、平成４年６月２日（寄託日）より独立行政法人産業技術総合研究所特許生物寄託センターにＦＥＲＭ Ｐ−１２９８１として寄託されている。また、リオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株は、それぞれＬｙｏｐｈｙｌｌｕｍ ｕｌｍａｒｉｕｍ Ｌｕ１−１７２、Ｌｙｏｐｈｙｌｌｕｍ ｕｌｍａｒｉｕｍ Ｌｕ１−１７３、Ｌｙｏｐｈｙｌｌｕｍ ｕｌｍａｒｉｕｍ Ｌｕ１−１７４、及びＬｙｏｐｈｙｌｌｕｍ ｕｌｍａｒｉｕｍ Ｌｕ１−１８１と表示され、ブダペスト条約の下、日本国茨城県つくば市東１丁目１番地１ 中央第６（郵便番号３０５−８５６６）独立行政法人産業技術総合研究所特許生物寄託センターに平成１４年（２００２年）４月２５日（原寄託日）にそれぞれ受託番号ＦＥＲＭ ＢＰ−８３５４、ＦＥＲＭ ＢＰ−８３５５、ＦＥＲＭ ＢＰ−８３５６、ＦＥＲＭ ＢＰ−８３５７として寄託されている。 There are no particular limitations on the type and strain of the mushroom in the present invention, but examples of the type of mushroom include shiitake, enokitake, oyster mushroom, nameko, maitake, riophyllum urumarium, and bamboo shimeji. The mushroom strain may be a commercially available strain, a tissue isolate from a wild fruiting body, or a strain bred by a method such as selection, mating, cell fusion, or gene recombination. For example, Riophyllum ulmarium K-0259, which is a lyophilum ulmarium strain, and lyophilum ulmarium Lu1-172 strain, Lu1-173 strain, Lu1-174 strain, and Lu1-181 strain, which are new strains described later, can be used. Riophyllum ulmarium K-0259 is displayed as Lyophyllum ulmarium K-0259, and has been deposited as FERM P-12981 at the National Institute of Advanced Industrial Science and Technology Patent Organism Depositary on June 2, 1992 (Deposit Date). Yes. In addition, the lyophilum ulmarium Lu1-172 strain, the Lu1-173 strain, the Lu1-174 strain, and the Lu1-181 strain are respectively Lyophyllum ulmarium Lu1-172, Under the Budapest Treaty, under the Budapest Treaty, 1st 1-1 Higashi 1-chome, Tsukuba, Ibaraki, Japan, Central 6 (postal code 305-866), the National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center in 2002 ) As of April 25 (original deposit date), accession numbers FERM BP-8354, FERM BP-8355, FERM BP-8356, FERM BP-8357 It has been committed.
一般にきのこにおいては同じ種に属する菌株であっても、菌株間で特性が大きく異なることが知られている。本発明者らはリオフィラム ウルマリウムにおいても公知菌株より、１種類以上のセリ科植物由来の処理物からなる基材を含有する培養基で良好な子実体形成能を有する菌株が存在するはずであるとの考えに立ち、鋭意育種検討を続けた。その結果、従来より用いられている人工栽培方法により良好な子実体形成能を有し、特に１種類以上のセリ科植物由来の処理物からなる基材を含有する培養基で人工栽培したときに、良好な子実体形成能を有するリオフィラム ウルマリウム新菌株を見出した。 In general, it is known that the characteristics of mushrooms are greatly different among strains even if they belong to the same species. In the lyophilum urumarium, the inventors of the present invention should also have a strain having a good fruiting body forming ability in a culture medium containing a base material made of a processed product derived from one or more species of the family Aceraceae. Based on his thoughts, he continued to study diligently. As a result, it has a good fruiting body forming ability by the conventionally used artificial cultivation method, and particularly when artificially cultivated in a culture medium containing a base material made of a processed product derived from one or more celery family plants, A new strain of liophyllum ulmarium with good fruiting ability was found.
以下、本発明におけるリオフィラム ウルマリウム新菌株について詳しく説明する。新菌株は交配育種により得た。交配育種は常法により以下のごとく行った。
本発明者らが保有するリオフィラム ウルマリウム菌株を針葉樹オガクズと米糠を混合した培養基を用いて子実体を発生させ、得られた子実体から胞子を回収し、発芽させて一核菌糸を計５０株単離し、総当たり交配により約１０００株の交配株を得た。
この約１０００株についてアシタバ青汁の搾汁粕を含有する培養基を用いて人工栽培試験を行い、良好な子実体形成能を有する株４株を選抜し、リオフィラム ウルマリウム Ｌu１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株と命名した。
Hereinafter, the new lyophilum ulmarium strain in the present invention will be described in detail. The new strain was obtained by cross breeding. Cross breeding was performed by the conventional method as follows.
The phyllophyllum ulmarium strain possessed by the present inventors is used to generate fruiting bodies using a culture medium in which coniferous sawdust and rice bran are mixed, and spores are recovered from the resulting fruiting bodies and germinated to obtain a total of 50 mononuclear mycelia. Separated, and about 1000 hybrids were obtained by brute force mating.
About 1000 strains were subjected to an artificial cultivation test using a culture medium containing Ashitaba Aojiru squeezed rice cake, 4 strains having good fruiting body forming ability were selected, and Riophylamumumarium Lu1-172 strain, Lu1-173 were selected. Strains, Lu1-174 strain, and Lu1-181 strain.
次にリオフィラム ウルマリウム Ｌｕ１−１７２株の諸性質を示す。
（１）麦芽エキス寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は４１ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体に密な菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は３７ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体を気菌糸が覆い、コロニー中央部付近がやや黄色、他は白色となる。
（３）ツアペック・ドックス寒天培地（２５℃）での生育状態
１０日目で小程度の生育、コロニー径は２５ｍｍ。菌糸は樹状に伸長し、極めて希薄、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育。菌糸は樹状で白色、希薄である。
（４）サブロー寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４２ｍｍ。白色で綿状の密な菌糸、気菌糸やや多い。２０日目でシャーレ全体に菌糸が生育し、気菌糸極めて多く、菌糸は綿状で白色である。
（５）オートミール寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４７ｍｍ。菌糸はよく分枝して伸び、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育、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℃）での生育状態
１０日目で小程度の生育、コロニー径は２３ｍｍ。菌糸は白色で直線状に伸び、放射繊維状のコロニーを形成する。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４２ｍｍ。菌糸は白色密で気菌糸を多量に生じる。マット状に生育。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色であるが、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℃）での生育状態
０．５％没食子酸添加バレイショ・ブドウ糖寒天培地を使用。１０日目小程度の生育、コロニー径は１９ｍｍ。菌糸は白色で短くマット状、気菌糸は少ない。培地は褐変、褐変半径は３２ｍｍ。２０日目では中程度の化育、コロニー径は３８ｍｍ、培地全体が褐変。種菌の新旧により著しく生育速度に差（約２倍）が生じる。
（９）最適生育温度
ＰＧＹ寒天培地（グルコース２％、ポリペプトン０．２％、酵母エキス０．２、ＫＨ2ＰＯ4０．５％、ＭｇＳＯ4・７Ｈ2Ｏ０．５％、寒天２％）に直径６ｍｍの円盤状種菌を接種し、各温度でそれぞれ培養して、１２日後に各コロニー径を測定したところ、最適生育温度は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（ＰＧＹ寒天培地から寒天を除いたもの）６０ｍｌずつを１００ｍｌ容の三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に直径６ｍｍの円盤状種菌を接種し、２５℃で１５日間静置培養後、菌体の乾燥重量を測定したところ、最適生育ｐＨは７〜８であった。また、本菌株の生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
Next, various properties of the lyophilum ulmarium Lu1-172 strain are shown.
(1) Growth state on malt extract agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 41 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, dense hyphae are formed on the entire surface. Mycelium is white.
(2) Growth state on potato / glucose agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 37 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, the entire surface is covered with aerial hyphae, the central part of the colony is slightly yellow, and the others are white.
(3) Growth condition on the Tuapec-Docx agar medium (25 ° C.) Small growth on the 10th day, colony diameter is 25 mm. Mycelium grows in a dendritic form and is extremely dilute and has few aerial hyphae. On the 20th day, hyphae grew on the entire petri dish. The mycelium is dendritic, white and dilute.
(4) Growth state on Sabouraud agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 42 mm. White, cotton-like dense hyphae, aerial hyphae are slightly more common. On the 20th day, hyphae grow on the entire petri dish, and there are extremely aerial hyphae.
(5) Growth state on oatmeal agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 47 mm. The hyphae are well branched and stretched, and there are few aerial hyphae. On the 20th day, hyphae grow on the entire petri dish, and a large amount of cotton-like air hyphae is produced. Mycelium is white.
(6) Growth state on synthetic mucor agar medium (25 ° C.) Small growth on day 10, colony diameter is 23 mm. The mycelium is white and extends linearly to form a radial fibrous colony. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. Mycelium is white.
(7) Growth state on YpSs agar medium (25 ° C.) Vigorous growth on day 10, colony diameter is 42 mm. The mycelium is white dense and produces a large amount of aerial hyphae. Grows on a mat. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. The mycelium is white, but the medium turns yellow.
(8) Growth state in phenol oxidase assay medium (25 ° C.) A potato / glucose agar medium supplemented with 0.5% gallic acid is used. Small growth on day 10, colony diameter is 19 mm. The mycelium is white, short and mat-like, and there are few aerial hyphae. The medium is browned and the browning radius is 32 mm. On the 20th day, medium growth, colony diameter was 38 mm, and the whole medium was browned. There is a marked difference in growth rate (about twice) depending on the age of the inoculum.
(9) Optimal growth temperature PGY agar medium (glucose 2%, polypeptone 0.2%, yeast extract 0.2, KH 2 PO 4 0.5%, MgSO 4 .7H 2 O 0.5%, agar 2%) When inoculated with a discoid inoculum having a diameter of 6 mm, cultured at each temperature, and measured each colony diameter after 12 days, the optimum growth temperature was around 25 ° C. Moreover, it hardly grew at 5 degreeC and 35 degreeC.
(10) Optimum growth pH
Dispense 60 ml of PGY liquid medium (PGY agar medium without agar) into 100 ml Erlenmeyer flasks, adjust each pH with acid or alkali, and inoculate a 6 mm diameter discoid inoculum. 25 When the dry weight of the microbial cells was measured after standing culture at 15 ° C. for 15 days, the optimum growth pH was 7-8. Moreover, the pH range in which this strain can grow was pH 3.5-10.
次にリオフィラム ウルマリウム Ｌｕ１−１７３株の諸性質を示す。
（１）麦芽エキス寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は４５ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体に密な菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は４３ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体を気菌糸が覆い、コロニー中央部付近がやや黄色、他は白色となる。
（３）ツアペック・ドックス寒天培地（２５℃）での生育状態
１０日目で小程度の生育、コロニー径は２３ｍｍ。菌糸は樹状に伸長し、極めて希薄・気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育。菌糸は樹状で白色、希薄である。
（４）サブロー寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４３ｍｍ。白色で綿状の密な菌糸、気菌糸やや多い。２０日目でシャーレ全体に菌糸が生育し、気菌糸極めて多く、菌糸は綿状で白色である。
（５）オートミール寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は５２ｍｍ。菌糸はよく分枝して伸び、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育、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℃）での生育状態
１０日目で小程度の生育、コロニー径は２１ｍｍ。菌糸は白色で直線状に伸び、放射繊維状のコロニーを形成する。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４０ｍｍ。菌糸は白色、密で気菌糸を多量に生じる。マット状に生育。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色であるが、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℃）での生育状態
０．５％没食子酸添加バレイショ・ブドウ糖寒天培地を使用。１０日目小程度の生育、コロニー径は２２ｍｍ。菌糸は白色で短くマット状、気菌糸は少ない。培地は褐変、褐変半径は３０ｍｍ。２０日目では中程度の生育、コロニー径は３６ｍｍ、培地全体が褐変。種菌の新旧により著しく生育速度に差（約２倍）が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径６ｍｍの円盤状種菌を接種し、各温度でそれぞれ培養して、１２日後に各コロニー径を測定したところ、最適生育温度は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６０ｍｌずつを１００ｍｌ容の三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に直径６ｍｍの円盤状種菌を接種し、２５℃で１５日間静置培養後、菌体の乾燥重量を測定したところ、最適生育ｐＨは７〜８であった。また、本菌株の生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
Next, various properties of the lyophilum ulmarium Lu1-173 strain are shown.
(1) Growth state on malt extract agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 45 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, dense hyphae are formed on the entire surface. Mycelium is white.
(2) Growth state on potato / glucose agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 43 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, the entire surface is covered with aerial hyphae, the central part of the colony is slightly yellow, and the others are white.
(3) Growth condition on Tappek Docs agar medium (25 ° C.) Small growth on day 10, colony diameter is 23 mm. The mycelium grows in a dendritic form and is extremely dilute and few aerial hyphae. On the 20th day, hyphae grew on the entire petri dish. The mycelium is dendritic, white and dilute.
(4) Growth state on Sabouraud agar medium (25 ° C.) Vigorous growth on 10th day, colony diameter is 43 mm. White, cotton-like dense hyphae, aerial hyphae are slightly more common. On the 20th day, hyphae grow on the entire petri dish, and there are extremely aerial hyphae, and the hyphae are cottony and white.
(5) Growth state on oatmeal agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 52 mm. The hyphae are well branched and stretched, and there are few aerial hyphae. On the 20th day, hyphae grow on the entire petri dish, and a large amount of cotton-like air hyphae is produced. Mycelium is white.
(6) Growth state on synthetic mucor agar medium (25 ° C.) Small growth on day 10, colony diameter is 21 mm. The mycelium is white and extends linearly to form a radial fibrous colony. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. Mycelium is white.
(7) Growth state on YpSs agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 40 mm. The mycelium is white, dense and produces a large amount of aerial hyphae. Grows on a mat. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. The mycelium is white, but the medium turns yellow.
(8) Growth state in phenol oxidase assay medium (25 ° C.) A potato / glucose agar medium supplemented with 0.5% gallic acid is used. Small growth on day 10, colony diameter is 22 mm. The mycelium is white, short and mat-like, and there are few aerial hyphae. The medium is browned and the browning radius is 30 mm. On the 20th day, medium growth, the colony diameter was 36 mm, and the whole medium was browned. There is a marked difference in growth rate (about twice) depending on the age of the inoculum.
(9) Optimal growth temperature When a PGY agar medium was inoculated with a discoid inoculum having a diameter of 6 mm, cultured at each temperature, and each colony diameter was measured after 12 days, the optimal growth temperature was around 25 ° C. Moreover, it hardly grew at 5 degreeC and 35 degreeC.
(10) Optimal growth pH
Dispense 60 ml each of PGY liquid medium into a 100 ml Erlenmeyer flask, adjust to each pH with acid or alkali, inoculate with 6 mm diameter discoid inoculum, incubate statically at 25 ° C. for 15 days, and then cells When the dry weight was measured, the optimum growth pH was 7-8. Moreover, the pH range in which this strain can grow was pH 3.5-10.
次にリオフィラム ウルマリウム Ｌｕ１−１７４株の諸性質を示す。
（１）麦芽エキス寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は４３ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体に密な菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は４１ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体を気菌糸が覆い、コロニー中央部付近がやや黄色、他は白色となる。
（３）ツアペック・ドックス寒天培地（２５℃）での生育状態
１０日目で小程度の生育、コロニー径は２５ｍｍ。菌糸は樹状に伸長し、極めて希薄、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育。菌糸は樹状で白色、希薄である。
（４）サブロー寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４５ｍｍ。白色で綿状の密な菌糸、気菌糸やや多い。２０日目でシャーレ全体に菌糸が生育し、気菌糸極めて多く、菌糸は綿状で白色である。
（５）オートミール寒天培地（２５℃）での生育状態
1０日目で旺盛な生育、コロニー径は５０ｍｍ。菌糸はよく分枝して伸び、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育、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℃）での生育状態
１０日目で小程度の生育、コロニー径は１９ｍｍ。菌糸は白色で直線状に伸び、放射繊維状のコロニーを形成する。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４２ｍｍ。菌糸は白色、密で気菌糸を多量に生じて、マット状に生育。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色であるが、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℃）での生育状態
０．５％没食子酸添加バレイショ・ブドウ糖寒天培地を使用。１０日目小程度の生育、コロニー径は２０ｍｍ。菌糸は白色で短くマット状、気菌糸は少ない。培地は褐変・褐変半径は３７ｍｍ。２０日目では中程度の生育、コロニー径は３８ｍｍ、培地全体が褐変。種菌の新旧により著しく生育速度に差（約２倍）が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径６ｍｍの円盤状種菌を接種し、各温度でそれぞれ培養して、１２日後に各コロニー径を測定したところ、最適生育温度は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６０ｍｌずつを１００ｍｌ容の三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に直径６ｍｍの円盤状種菌を接種し、２５℃で１５日間静置培養後、菌体の乾燥重量を測定したところ、最適生育ｐＨは７〜８であった。また、本菌株の生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
Next, various properties of the lyophilum ulmarium Lu1-174 strain are shown.
(1) Growth state on malt extract agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 43 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, dense hyphae are formed on the entire surface. Mycelium is white.
(2) Growth state on potato / glucose agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 41 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, the entire surface is covered with aerial hyphae, the central part of the colony is slightly yellow, and the others are white.
(3) Growth condition on the Tuapec-Docx agar medium (25 ° C.) Small growth on the 10th day, colony diameter is 25 mm. Mycelium grows in a dendritic form and is extremely dilute and has few aerial hyphae. On the 20th day, hyphae grew on the entire petri dish. The mycelium is dendritic, white and dilute.
(4) Growth state on Sabouraud agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 45 mm. White, cotton-like dense hyphae, aerial hyphae are slightly more common. On the 20th day, hyphae grow on the entire petri dish, and there are extremely aerial hyphae, and the hyphae are cottony and white.
(5) Growth condition on oatmeal agar medium (25 ° C)
Vigorous growth on day 10, colony diameter is 50 mm. The hyphae are well branched and stretched, and there are few aerial hyphae. On the 20th day, hyphae grow on the entire petri dish, and a large amount of cotton-like air hyphae is produced. Mycelium is white.
(6) Growth condition on synthetic mucor agar medium (25 ° C.) Small growth on day 10, colony diameter is 19 mm. The mycelium is white and extends linearly to form a radial fibrous colony. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. Mycelium is white.
(7) Growth state on YpSs agar medium (25 ° C.) Vigorous growth on day 10, colony diameter is 42 mm. The mycelium is white, dense and produces a large amount of aerial hyphae and grows in a mat shape. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. The mycelium is white, but the medium turns yellow.
(8) Growth state in phenol oxidase assay medium (25 ° C.) A potato / glucose agar medium supplemented with 0.5% gallic acid is used. Small growth on day 10, colony diameter is 20 mm. The mycelium is white, short and mat-like, and there are few aerial hyphae. The medium has a browning / browning radius of 37 mm. On the 20th day, medium growth, the colony diameter was 38 mm, and the whole medium was browned. There is a marked difference in growth rate (about twice) depending on the age of the inoculum.
(9) Optimal growth temperature When a PGY agar medium was inoculated with a discoid inoculum having a diameter of 6 mm, cultured at each temperature, and each colony diameter was measured after 12 days, the optimal growth temperature was around 25 ° C. Moreover, it hardly grew at 5 degreeC and 35 degreeC.
(10) Optimum growth pH
Dispense 60 ml each of PGY liquid medium into a 100 ml Erlenmeyer flask, adjust to each pH with acid or alkali, inoculate with 6 mm diameter discoid inoculum, incubate statically at 25 ° C. for 15 days, and then cells When the dry weight was measured, the optimum growth pH was 7-8. Moreover, the pH range in which this strain can grow was pH 3.5-10.
次にリオフィラム ウルマリウム Ｌｕ１−１８１株の諸性質を示す。
（１）麦芽エキス寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、コロニー径は４５ｍｍ。白色で密な菌糸、気菌糸を多量に生じる。１０日目でシャーレ全体に菌糸が生育。２０日目には表面全体に密な菌糸を生じる。菌糸は白色。
（２）バレイショ・ブドウ糖寒天培地（２５℃）での生育状態
７日目で旺盛な生育で、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日目でシャーレ全体に菌糸が生育。２０日目には表面全体を気菌糸が覆い、コロニー中央部付近がやや黄色、他は白色となる。
（３）ツアペック・ドックス寒天培地（２５℃）での生育状態
１０日目で小程度の生育、コロニー径は２７ｍｍ。菌糸は樹状に伸長し、極めて希薄、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育。菌糸は樹状で白色、希薄である。
（４）サブロー寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４６ｍｍ。白色で綿状の密な菌糸、気菌糸やや多い。２０日目でシャーレ全体に菌糸が生育し、気菌糸極めて多く、菌糸は綿状で白色である。
（５）オートミール寒天培地（２５℃）での生育状態
１０日目で旺盛な生育、コロニー径は４８ｍｍ。菌糸はよく分枝して伸び、気菌糸は少ない。２０日目でシャーレ全体に菌糸が生育、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℃）での生育状態
１０日目で小程度の生育、コロニー径は２０ｍｍ。菌糸は白色で直線状に伸び、放射繊維状のコロニーを形成する。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色。
（７）ＹｐＳｓ寒天培地（２５℃）での生育状態
1０日目で旺盛な生育、コロニー径は４４ｍｍ。菌糸は白色、密で気菌糸を多量に生じる。マット状に生育。２０日目で菌糸はシャーレ全体に生育し、気菌糸を多量に生じる。菌糸は白色であるが、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℃）での生育状態
０．５％没食子酸添加バレイショ・ブドウ糖寒天培地を使用。１０日目小程度の生育、コロニー径は１７ｍｍ。菌糸は白色で短くマット状、気菌糸は少ない。培地は褐変、褐変半径は３２ｍｍ。２０日目では中程度の生育、コロニー径は３６ｍｍ、培地全体が褐変。種菌の新旧により著しく生育速度に差（約２倍）が生じる。
（９）最適生育温度
ＰＧＹ寒天培地に直径６ｍｍの円盤状種菌を接種し、各温度でそれぞれ培養して、１２日後に各コロニー径を測定したところ、最適生育温度は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６０ｍｌずつを１００ｍｌ容の三角フラスコに分注して殺菌し、酸又はアルカリで各ｐＨに調整後に直径６ｍｍの円盤状種菌を接種し、２５℃で１５日間静置培養後、菌体の乾燥重量を測定したところ、最適生育ｐＨは７〜８であった。また、本菌株の生育可能なｐＨ範囲は、ｐＨ３．５〜１０であった。
Next, various properties of the lyophilum ulmarium Lu1-181 strain are shown.
(1) Growth state on malt extract agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 45 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, dense hyphae are formed on the entire surface. Mycelium is white.
(2) Growth state on potato / glucose agar medium (25 ° C.) Vigorous growth on day 7, colony diameter is 40 mm. A large amount of white and dense hyphae and aerial hyphae are produced. On the 10th day, hyphae grew on the entire petri dish. On the 20th day, the entire surface is covered with aerial hyphae, the central part of the colony is slightly yellow, and the others are white.
(3) Growth condition on Zappec Doc agar medium (25 ° C.) Small growth on the 10th day, colony diameter is 27 mm. Mycelium grows in a dendritic form and is extremely dilute and has few aerial hyphae. On the 20th day, hyphae grew on the entire petri dish. The mycelium is dendritic, white and dilute.
(4) Growth state on Sabouraud agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 46 mm. White, cotton-like dense hyphae, aerial hyphae are slightly more common. On the 20th day, hyphae grow on the entire petri dish, and there are extremely aerial hyphae.
(5) Growth state on oatmeal agar medium (25 ° C.) Vigorous growth on the 10th day, colony diameter is 48 mm. The hyphae are well branched and stretched, and there are few aerial hyphae. On the 20th day, hyphae grow on the entire petri dish, and a large amount of cotton-like air hyphae is produced. Mycelium is white.
(6) Growth state on synthetic mucor agar medium (25 ° C.) Small growth on day 10, colony diameter is 20 mm. The mycelium is white and extends linearly to form a radial fibrous colony. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. Mycelium is white.
(7) Growth state on YpSs agar medium (25 ° C)
Vigorous growth on day 10, colony diameter is 44 mm. The mycelium is white, dense and produces a large amount of aerial hyphae. Grows on a mat. On the 20th day, the mycelium grows throughout the petri dish and produces a large amount of aerial hyphae. The mycelium is white, but the medium turns yellow.
(8) Growth state in phenol oxidase assay medium (25 ° C.) A potato / glucose agar medium supplemented with 0.5% gallic acid is used. Small growth on day 10, colony diameter is 17 mm. The mycelium is white, short and mat-like, and there are few aerial hyphae. The medium is browned and the browning radius is 32 mm. On the 20th day, medium growth, the colony diameter was 36 mm, and the whole medium was brown. There is a marked difference in growth rate (about twice) depending on the age of the inoculum.
(9) Optimal growth temperature When a PGY agar medium was inoculated with a discoid inoculum having a diameter of 6 mm, cultured at each temperature, and each colony diameter was measured after 12 days, the optimal growth temperature was around 25 ° C. Moreover, it hardly grew at 5 degreeC and 35 degreeC.
(10) Optimum growth pH
Dispense 60 ml each of PGY liquid medium into a 100 ml Erlenmeyer flask, adjust to each pH with acid or alkali, inoculate with 6 mm diameter discoid inoculum, incubate statically at 25 ° C. for 15 days, and then cells When the dry weight was measured, the optimum growth pH was 7-8. Moreover, the pH range in which this strain can grow was pH 3.5-10.
次に、リオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株と他のリオフィラム ウルマリウム菌株との異同判定として、両菌糸が持つ性因子が異なれば、その菌糸は互いに異なる菌糸であるという菌類分類学的事実に基づき、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。
供試した菌株としてリオフィラム ウルマリウム ＩＦＯ９６３７株、ＩＦＯ３０５２５株、ＩＦＯ３０７７５株、リオフィラム ウルマリウム Ｍ−８１７１株（ＦＥＲＭ ＢＰ−１４１５）、及びリオフィラム ウルマリウム Ｋ−０２５９株（ＦＥＲＭＰ−１２９８１）を用いた。ここでリオフィラム ウルマリウム Ｍ−８１７１株、及びリオフィラム ウルマリウム Ｋ−０２５９株はかつて本発明者らによって育種されたリオフィラム ウルマリウム菌株である。
これらの各菌株の二核菌糸を ＰＧＹ寒天培地で増殖させ、３ｍｍ×３ｍｍ×３ｍｍのブロックとして切り出し、それぞれをＰＧＹ寒天平板培地の中央部に、リオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株の２核菌糸ブロック（３ｍｍ×３ｍｍ×３ｍｍ）と対峙して約２cｍ間隔で植菌し、２５℃、１４日間培養後、両コロニー境界部に帯線が生じるか否かを判定した。帯線を生じた場合を＋、生じなかった場合を−として、この結果を表１に示す。
Next, as a difference determination between the lyophilum ulmarium Lu1-172 strain, the Lu1-173 strain, the Lu1-174 strain, and the Lu1-181 strain and other lyophilum ulmarium strains, Based on the fungal taxonomic fact that the mycelia are different from each other, the difference in sex factors was examined by counterculture on an agar medium.
As strains tested, Riophylam ulmarium IFO9637 strain, IFO30525 strain, IFO30775 strain, Riofilum ulmarium M-8171 strain (FERM BP-1415), and Riofilum ulmarium K-0259 strain (FERMP-12981) were used. Here, lyophilum ulmarium M-8171 strain and lyophilum ulmarium K-0259 strain are lyophilum ulmarium strains that were once bred by the present inventors.
The dinuclear mycelium of each of these strains was grown on a PGY agar medium, cut out as a 3 mm × 3 mm × 3 mm block, and each of them was placed at the center of the PGY agar plate medium with the lyophilum ulmarium Lu1-172 strain, the Lu1-173 strain, the Lu1 -174 and Lu1-181 strains of binuclear hyphae blocks (3 mm x 3 mm x 3 mm) are inoculated at about 2 cm intervals, and after culturing at 25 ° C for 14 days, a band appears at the boundary between both colonies It was determined whether or not. The results are shown in Table 1 where + is the case where the band is generated and − is the case where the band is not generated.
表１に示すように、リオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−1８１株はそれぞれ他の菌株との対峙培養ですべて帯線を生じた。このことからリオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株は新しい菌株であることは明白である。
また、上記に示した本発明の新菌株であるリオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株は、菌床人工栽培において短い培養日数、例えば４０日〜７０日程度の培養日数でも良好な子実体形成能を有するという特徴をも有しており、特に好適にはセリ科植物由来の処理物からなる基材を含有する培養基で培養日数が短く、人工栽培に適した菌株である。
As shown in Table 1, Riofilum ulmarium Lu1-172 strain, Lu1-173 strain, Lu1-174 strain, and Lu1-181 strain all showed band lines in counter-culture with other strains. From this, it is clear that Riophyllum ulmarium Lu1-172 strain, Lu1-173 strain, Lu1-174 strain, and Lu1-181 strain are new strains.
In addition, the new strains of the present invention shown above, Riophyrum ulmarium Lu1-172 strain, Lu1-173 strain, Lu1-174 strain, and Lu1-181 strain, have a short culture days, for example, 40 days to It also has a feature that it has a good fruiting body forming ability even in about 70 days of culture. Particularly preferably, the culture medium containing a base material made of a processed product derived from a celery family has a short culture day, and is artificial. It is a strain suitable for cultivation.
以上説明したように本発明の新菌株として、例えばリオフィラム ウルマリウム Ｌｕ１−１７２株、Ｌｕ１−１７３株、Ｌｕ１−１７４株、及びＬｕ１−１８１株が挙げられるが、前記菌株と同様にセリ科植物由来の処理物からなる基材を含有する培養基で良好な子実体形成能を有する菌株はすべて本発明に属するものである。 As described above, examples of the new strain of the present invention include the lyophilum urumalium Lu1-172 strain, the Lu1-173 strain, the Lu1-174 strain, and the Lu1-181 strain. All strains having a good fruiting body forming ability in a culture medium containing a substrate made of a treated product belong to the present invention.
以下に本発明を実施例により更に具体的に説明するが、本発明は、以下の実施例の範囲のみに限定されるものではない。 The present invention will be described more specifically with reference to the following examples. However, the present invention is not limited to the scope of the following examples.
実施例１
アシタバ搾汁粕（アシタバ青汁の絞り粕）及びコーンコブ〔タカラ物産（株）製〕を乾燥重量でそれぞれ６５ｇずつ、よく混合して水分含有率６８％となるように調製し、これをポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕に詰めてビン口中央より下方に向かい直径１ｃｍの穴をあけた後、キャップで打栓した。これを１２０℃、６０分間高圧殺菌して培養基を調製した。該培養基にきのことしてリオフィラム ウルマリウム Ｋ−０２５９、ＩＦＯ９６３７、及びＩＦＯ３０５２５株の固体種菌を接種し、２５℃の条件下で３０日培養して培養菌糸を得た後、更に５０日培養して子実体発生基を得た。該子実体発生基の上部菌糸層１ｃｍを中央部を残して除去し（菌掻き）、水道水２０ｍlを加えて充分に吸水させた後、余剰の水を除いて１５℃、湿度９０％以上の条件下でＫ−０２５９株とＩＦＯ３０５２５株は１０日間、ＩＦＯ９６３７株は１３日間培養して子実体原基を形成させ、光を照射して、更にＫ−０２５９株とＩＦＯ３０５２５株は１２日間、ＩＦＯ９６３７株は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Ｋ−０２５９株が７５ｇ、ＩＦＯ３０５２５株が１０５ｇ、ＩＦＯ９６３７株が７６ｇであり、それぞれ高品質なものであった。
Example 1
Ashitaba squeezed rice cake (Ashitaba green juice squeezed rice cake) and corn cob (manufactured by Takara Bussan Co., Ltd.) were prepared by mixing 65 g each in dry weight so that the water content was 68%. After filling a 850 ml bottle [trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.] and making a hole with a diameter of 1 cm downward from the center of the bottle mouth, it was stoppered with a cap. This was pasteurized at 120 ° C. for 60 minutes to prepare a culture medium. Inoculated with solid inoculum of Riophylam ulmarium K-0259, IFO9637, and IFO30525 strains to the culture medium, and cultured for 30 days at 25 ° C. to obtain cultured mycelia, followed by further cultivation for 50 days. The group was obtained. After removing 1 cm of the upper mycelium layer of the fruiting body leaving the central part (fungal scraping), adding 20 ml of tap water to sufficiently absorb water, and removing excess water, the temperature is 15 ° C. and the humidity is 90% or more. Under the conditions, the K-0259 strain and the IFO 30525 strain were cultured for 10 days, the IFO 9637 strain was cultured for 13 days to form fruiting body primordia, irradiated with light, and further the K-0259 strain and the IFO 30525 strain for 12 days, the IFO 9637 strain. Continued to culture for 14 days to obtain fruiting bodies. The yield of the fruiting bodies obtained was 75 g for the K-0259 strain, 105 g for the IFO 30525 strain, and 76 g for the IFO 9637 strain, each of high quality.
比較例１
コーンコブを使用せず、アシタバ搾汁粕のみを1３０ｇ用いる点と子実体原基形成にＫ−０２５９株とＩＦＯ３０５２５株は１１日間、ＩＦＯ９６３７株は１４日間、子実体を得るまでに更にＫ−０２５９株とＩＦＯ３０５２５株は１３日間、ＩＦＯ９６３７株は１４日間培養を続けた点を除いては実施例１と同様に行って子実体を得た。得られた子実体の収量はＫ−０２５９株が５５ｇ、ＩＦＯ３０５２５株が４５ｇ、ＩＦＯ９６３７株が３４ｇであり、アシタバ搾汁粕単独より、アシタバ搾汁粕とコーンコブを用いた培養基の方が優れていた。また、栽培日数もアシタバ搾汁粕とコーンコブを用いた方が短かった。
Comparative Example 1
Without using corn cob, use only 130g of Ashitaba squeezed rice cake, and K-0259 strain and IFO 30525 strain for 11 days for fruit body primordium formation, 14 days for IFO 9637 strain, and K-0259 strain until fruit body is obtained In the same manner as in Example 1, except that IFO30525 strain was cultured for 13 days and IFO9637 strain was cultured for 14 days, fruiting bodies were obtained. The yield of the fruiting body obtained was 55 g for K-0259 strain, 45 g for IFO 30525 strain and 34 g for IFO 9637 strain, and the culture medium using Ashitaba squeezed rice cake and corn cob was superior to Ashitaba squeezed rice cake alone. . In addition, the cultivation days were shorter when using Ashitaba squeezed rice cake and corn cob.
実施例２
アシタバ搾汁粕、針葉樹オガクズ〔（有）唐澤商運製〕、及び米糠を乾燥重量で、それぞれ４６ｇ、４６ｇ、及び７４ｇをよく混合して水分含有率６８％となるように調製し、これを用いる以外は実施例１と同様にして子実体を得た。
得られた子実体の収量はＫ−０２５９株が８７ｇ、ＩＦＯ３０５２５株が１４９ｇ、ＩＦＯ９６３７株が９７ｇであり、それぞれ高品質なものであった。
Example 2
Ashitaba squeezed rice cake, coniferous sawdust (produced by Karasawa Shoun Co., Ltd.), and rice bran were prepared by mixing 46 g, 46 g, and 74 g in dry weight to a moisture content of 68%. A fruiting body was obtained in the same manner as in Example 1 except that was used.
The yields of the fruiting bodies obtained were 87 g for the K-0259 strain, 149 g for the IFO 30525 strain, and 97 g for the IFO 9637 strain, each of high quality.
比較例２
アシタバ搾汁粕を使用せず、針葉樹オガクズ〔（有）唐澤商運製〕９２ｇとコメヌカ７４ｇのみを用いる点と子実体原基形成にＫ−０２５９株とＩＦＯ３０５２５株は１１日間、ＩＦＯ９６３７株は１４日間、子実体を得るまでに更にＫ−０２５９株とＩＦＯ３０５２５株は１３日間、ＩＦＯ９６３７株は１４日間培養を続けた点を除いては実施例２と同様に行って子実体を得た。得られた子実体の収量はＫ−０２５９株が７１ｇ、ＩＦＯ３０５２５株が７２ｇ、ＩＦＯ９６３７株が７３ｇであり、アシタバ搾汁粕を用いた培養基の方が優れていた。
Comparative Example 2
Without using Ashitaba squeezed rice cake, K-0259 strain and IFO 30525 strain were used for 11 days, IFO 9637 strain was used for the formation of fruit body primordium and conifer tree sawdust (produced by Karasawa Trading Co., Ltd.) 92 g and rice bran 74 g For 14 days, fruit bodies were obtained in the same manner as in Example 2 except that K-0259 strain and IFO 30525 strain were further cultured for 13 days, and IFO 9637 strain was cultured for 14 days before the fruit bodies were obtained. The yield of the fruiting bodies obtained was 71 g for the K-0259 strain, 72 g for the IFO 30525 strain, and 73 g for the IFO 9637 strain, and the culture medium using Ashitaba squeezed koji was superior.
実施例３
アシタバ搾汁粕及びコーンコブ〔タカラ物産（株）製〕を乾燥重量でそれぞれ６５ｇずつ、並びにメタケイ酸アルミン酸マグネシウム〔商品名：ノイシリンＦＨ１、富士化学工業（株）製〕を新鮮重量で２．５ｇをよく混合して水分含有率６８％となるように調製し、きのことしてリオフィラム ウルマリウム Ｋ−０２５９株のみを用いる以外は実施例１と同様にして子実体を得た。得られた子実体の収量は１２９ｇであり、高品質なものであった。
Example 3
Ashitaba squeezed rice cake and corn cob (manufactured by Takara Bussan Co., Ltd.) each with a dry weight of 65 g, and magnesium metasilicate aluminate (trade name: Neusilin FH1, manufactured by Fuji Chemical Industry Co., Ltd.) with a fresh weight of 2.5 g Were mixed well to prepare a water content of 68%, and fruit bodies were obtained in the same manner as in Example 1 except that mushrooms were used only with Ryofilum ulmarium K-0259 strain. The yield of the fruiting body obtained was 129 g, which was high quality.
比較例３
アシタバ搾汁粕の代わりに針葉樹オガクズ〔（有）唐澤商運製〕を用いる点を除いては実施例３と同様に行って子実体を得た。得られた子実体の収量は９５ｇであり、アシタバ搾汁粕を用いた培養基の方が優れていた。
Comparative Example 3
A fruiting body was obtained in the same manner as in Example 3 except that coniferous sawdust [manufactured by Karasawa Trading Co., Ltd.] was used instead of Ashitaba squeezed rice cake. The yield of the fruiting body obtained was 95 g, and the culture medium using Ashitaba squeezed koji was superior.
実施例４
アシタバ搾汁粕及びコーンコブ〔タカラ物産（株）製〕を乾燥重量でそれぞれ６５ｇずつ、並びにメタケイ酸アルミン酸マグネシウム〔商品名：ノイシリンＦＨ１、富士化学工業（株）製〕を新鮮重量で２．５ｇをよく混合して水分含有率６８％となるように調製し、これをポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕に詰めてビン口中央より下方に向かい直径１ｃｍの穴をあけた後、キャップで打栓した。これを１２０℃、６０分間高圧殺菌して培養基を調製した。該培養基に本発明のリオフィラム ウルマリウム新菌株としてリオフィラム ウルマリウム Ｌｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８1株の固体種菌を接種し、２５℃の条件下で３０日培養して培養菌糸を得た後、更に４０日培養して子実体発生基を得た。該子実体発生基の上部菌糸層１ｃｍを中央部を残して除去し（菌掻き）、水道水２０ｍｌを加えて充分に吸水させた後、余剰の水を除いて１５℃、湿度９０％以上の条件下で１０日間培養して子実体原基を形成させ、光を照射して更に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Ｌｕ１−１７２株が１４０ｇ、Ｌｕ１−１７３株が１３８ｇ、Ｌｕ１−１７４株が１３３ｇ、Ｌｕ１−１８１株が１３２ｇであり、それぞれ高品質なものであった。
Example 4
Ashitaba squeezed rice cake and corn cob (manufactured by Takara Bussan Co., Ltd.) each with a dry weight of 65 g, and magnesium metasilicate aluminate (trade name: Neusilin FH1, manufactured by Fuji Chemical Industry Co., Ltd.) with a fresh weight of 2.5 g Was mixed well to prepare a water content of 68%, packed in a polypropylene 850 ml bottle [trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.] and directed downward from the center of the bottle mouth. After making a hole with a diameter of 1 cm, it was stoppered with a cap. This was pasteurized at 120 ° C. for 60 minutes to prepare a culture medium. The culture medium is inoculated with a solid inoculum of Riophylamumumarium Lu1-172, Lu1-173, Lu1-174, and Lu1-181 strain as a new strain of Lyophilum ulmarium of the present invention, cultured for 30 days at 25 ° C., and cultured hyphae Then, the fruit body generating group was obtained by further culturing for 40 days. After removing 1 cm of the upper mycelium layer of the fruiting body leaving the central part (fungal scraping) and adding 20 ml of tap water to sufficiently absorb water, the excess water is removed and the temperature is 15 ° C. and the humidity is 90% or more. The fruit body primordium was formed by culturing for 10 days under the conditions, and the fruit body was obtained by irradiating light and further culturing for 13 days. The yields of the fruiting bodies obtained were 140 g for the Lu1-172 strain, 138 g for the Lu1-173 strain, 133 g for the Lu1-174 strain, and 132 g for the Lu1-181 strain, which were high quality.
比較例４
リオフィラム ウルマリウム Ｋ−０２５９株を用いる点と、２５℃の条件下で３０日培養して培養菌糸を得た後更に５０日培養し子実体発生基を得た点を除いては実施例４と同様にして子実体を得た。得られた子実体の収量は１２９ｇであり、品質は良好であった。アシタバ搾汁粕を用いた培養基ではＬｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１株の方がリオフィラム ウルマリウム Ｋ−０２５９株より収量が優れ、短い培養日数で栽培が可能であった。
Comparative Example 4
Same as Example 4 except that liophyllum ulmarium K-0259 strain was used, and that cultured mycelia were obtained by culturing at 25 ° C. for 30 days and then cultured for 50 days to obtain fruiting body generating groups. I got a fruiting body. The yield of the fruit body obtained was 129 g, and the quality was good. In the culture medium using Ashitaba squeezed rice cake, the yields of Lu1-172, Lu1-173, Lu1-174, and Lu1-181 were superior to those of Riophyllum ulmarium K-0259 and could be cultivated in a short number of days. .
比較例５
リオフィラム ウルマリウム Ｋ−０２５９株、ＩＦＯ９６３７株、及びＩＦＯ３０５２５株を用いる点と、２５℃の条件下で３０日培養して培養菌糸を得た後更に４０日培養し子実体発生基を得た点を除いては実施例４と同様にして、Ｋ−０２５９株以外は子実体をえた。Ｋ−０２５９株は子実体を得られなかった。得られた子実体の収量はＩＦＯ３０５２５株が７８ｇ、ＩＦＯ９６３７株が７１ｇであり、品質は良好であった。アシタバ搾汁粕を用いた培養基で、同じ培養日数とした場合は、Ｌｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１株の方がリオフィラム ウルマリウム Ｋ−０２５９株、ＩＦＯ９６３７株、及びＩＦＯ３０５２５株より収量が優れていた。
Comparative Example 5
Except for the point of using lyophilum ulmarium K-0259 strain, IFO9637 strain, and IFO30525 strain and culturing for 30 days at 25 ° C. to obtain cultured mycelia and further culturing for 40 days. In the same manner as in Example 4, fruit bodies were obtained except for the strain K-0259. The fruit body of K-0259 strain was not obtained. The yield of the fruiting body obtained was 78 g for IFO 30525 strain and 71 g for IFO 9637 strain, and the quality was good. In the culture medium using Ashitaba squeezed lees, when the same culture days are used, the strains Lu1-172, Lu1-173, Lu1-174, and Lu1-181 are more lyophilum ulmarium K-0259, IFO9637, and IFO30525. The yield was better than the strain.
実施例５
針葉樹オガクズ〔（有）唐澤商運製〕５２ｇ、広葉樹オガクズ〔（有）トモエ物産製〕６９ｇ、及び米糠８６ｇを乾燥重量で計量し、よく混合して水分含有率６５％となるように調製し、これをポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕に詰めてビン口中央より下方に向かい直径１ｃｍの穴をあけた後、キャップで打栓した。これを１２０℃、６０分間高圧殺菌して培養基を調製した。該培養基にリオフィラム ウルマリウム菌株としてリオフィラム ウルマリウム Ｌｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１の固体種菌を接種し、２５℃の条件下で３０日培養して培養菌糸を得た後、更に４０日培養して子実体発生基を得た。該子実体発生基の上部菌糸層１ｃｍを中央部を残して除去し（菌掻き）、水道水２０ｍｌを加えて充分に吸水させた後、余剰の水を除いて１５℃、湿度９０％以上の条件下で１０日間培養して子実体原基を形成させ、光を照射して更に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Ｌｕ１−１７２株が１１５ｇ、Ｌｕ１−１７３株が１２６ｇ、Ｌｕ１−１７４株が１１５ｇ、Ｌｕ１−１８１株が１１５ｇであり、それぞれ高品質なものであった。
Example 5
Weigh 52 g of coniferous sawdust (made by Karasawa Trading Co., Ltd.), 69 g of hardwood sawdust (made by Tomoe Bussan), and 86 g of rice bran by dry weight, and mix well to prepare a water content of 65%. This was packed in a polypropylene 850 ml bottle (trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.), a hole having a diameter of 1 cm was formed downward from the center of the bottle mouth, and then plugged with a cap. This was pasteurized at 120 ° C. for 60 minutes to prepare a culture medium. After inoculating the culture medium with solid inoculum of lyophilum urmarium Lu1-172, Lu1-173, Lu1-174, and Lu1-181 as lyophilum ulmarium strain and culturing at 25 ° C. for 30 days to obtain cultured mycelia, Furthermore, the fruit body generating group was obtained by culturing for 40 days. After removing 1 cm of the upper mycelium layer of the fruiting body leaving the central part (fungal scraping) and adding 20 ml of tap water to sufficiently absorb water, the excess water is removed and the temperature is 15 ° C. and the humidity is 90% or more. The fruit body primordium was formed by culturing for 10 days under the conditions, and the fruit body was obtained by irradiating light and further culturing for 14 days. The yields of the fruiting bodies obtained were 115 g for the Lu1-172 strain, 126 g for the Lu1-173 strain, 115 g for the Lu1-174 strain, and 115 g for the Lu1-181 strain, which were high quality.
比較例６
リオフィラム ウルマリウム Ｋ−０２５９株を用いる点と、２５℃の条件下で３０日培養して培養菌糸を得た後更に５０日培養し子実体発生基を得た点を除いては実施例５と同様にして子実体を得た。得られた子実体の収量は１０８ｇであり、品質は良好であった。針葉樹オガクズ、広葉樹オガクズ及び米糠を用いた培養基ではＬｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１株の方がリオフィラム ウルマリウム Ｋ−０２５９株より収量が優れ、短い培養日数で栽培が可能であった。
Comparative Example 6
Same as Example 5 except that liophyllum ulmarium K-0259 strain was used, and that cultured mycelia were obtained by culturing at 25 ° C. for 30 days and then cultured for 50 days to obtain fruiting body generating groups. I got a fruiting body. The yield of the fruit body obtained was 108 g, and the quality was good. In the culture medium using coniferous sawdust, broad-leaved sawdust and rice bran, the Lu1-172, Lu1-173, Lu1-174, and Lu1-181 strains are superior in yield to the liofilum ulmarium K-0259 strain and can be cultivated in a shorter number of days. Met.
実施例６
ニンジン搾汁粕（ニンジン根茎の絞り粕）５２ｇ、針葉樹オガクズ〔（有）唐澤商運製〕５２ｇ、及び米糠８４ｇを乾燥重量で計量し、よく混合して水分含有率６５％となるように調製し、これをポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕に詰めてビン口中央より下方に向かい直径１ｃｍの穴をあけた後、キャップで打栓した。これを１２０℃、６０分間高圧殺菌して培養基を調製した。該培養基にきのことしてリオフィラム ウルマリウム Ｋ−０２５９株の固体種菌を接種し、２５℃の条件下で３０日培養して培養菌糸を得た後、更に５０日培養して子実体発生基を得た。該子実体発生基の上部菌糸層１ｃｍを中央部を残して除去し（菌掻き）、水道水２０ｍlを加えて充分に吸水させた後、余剰の水を除いて１５℃、湿度９０％以上の条件下で１３日間培養して子実体原基を形成させ、光を照射して更に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９２ｇであり、高品質なものであった。
Example 6
Weigh 52 g of carrot juice (carrot rhizome squeezed rice cake), 52 g of coniferous sawdust (produced by Karasawa Shoun Co., Ltd.), and 84 g of rice bran by dry weight, and mix well so that the water content is 65%. This was prepared, packed in a polypropylene 850 ml bottle (trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.), made a hole with a diameter of 1 cm from the center of the bottle mouth, and then plugged with a cap. . This was pasteurized at 120 ° C. for 60 minutes to prepare a culture medium. The culture medium was inoculated with a solid inoculum of Riophyllum ulmarium K-0259 strain, cultured for 30 days at 25 ° C. to obtain cultured mycelia, and further cultured for 50 days to obtain a fruiting body generating group. After removing 1 cm of the upper mycelium layer of the fruiting body leaving the central part (fungal scraping), adding 20 ml of tap water to sufficiently absorb water, and removing excess water, the temperature is 15 ° C. and the humidity is 90% or more. The fruit body primordium was formed by culturing for 13 days under the conditions, and the fruit body was obtained by irradiating light and further culturing for 15 days. The yield of the fruiting body obtained was 92 g, which was high quality.
比較例７
針葉樹オガクズとコメヌカを使用せず、ニンジン搾汁滓のみを1９２ｇ用いる点を除いては実施例６と同様の条件で栽培を行ったが、子実体は得られなかった。ニンジン搾汁粕単独より、ニンジン搾汁滓、針葉樹オガクズ、及び米糠を用いた培養基の方が優れていた。
Comparative Example 7
Cultivation was carried out under the same conditions as in Example 6 except that coniferous sawdust and rice bran were not used, and only 192 g of carrot juice was used, but no fruiting body was obtained. The culture medium using carrot juice koji, coniferous sawdust, and rice koji was superior to carrot juice koji alone.
実施例７
ニンジン搾汁粕（ニンジン根茎の絞り粕）３０ｇ、針葉樹オガクズ〔（有）唐澤商運製〕７０ｇ、及び米糠８１ｇを乾燥重量で計量し、よく混合して水分含有率６５％となるように調製し、これをポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕に詰めてビン口中央より下方に向かい直径１ｃｍの穴をあけた後、キャップで打栓した。これを１２０℃、６０分間高圧殺菌して培養基を調製した。該培養基にきのことしてリオフィラム ウルマリウム Ｌｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１株の固体種菌を接種し、２５℃の条件下で３０日培養して培養菌糸を得た後、更に４０日培養して子実体発生基を得た。該子実体発生基の上部菌糸層１ｃｍを中央部を残して除去し（菌掻き）、水道水２０ｍlを加えて充分に吸水させた後、余剰の水を除いて１５℃、湿度９０％以上の条件下でＬｕ１−１７２株は１２日間、Ｌｕ１−１７３株は１１日間、Ｌｕ１−１７４とＬｕ１−１８１株は１０日間培養して子実体原基を形成させ、光を照射して、更にＬｕ１−１７２株は１４日間、Ｌｕ１−１７３株は１３日間、Ｌｕ１−１７４株は１１日間、Ｌｕ１−１８１株は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Ｌｕ１−１７２株が１８１ｇ、Ｌｕ１−１７３株が１８９ｇ、Ｌｕ１−１７４株が１７３ｇ、Ｌｕ１−１８１株が１５９ｇであり、それぞれ高品質なものであった。
Example 7
Weigh 30 g of carrot juice (carrot rhizome squeezed potato), 70 g of coniferous sawdust (manufactured by Karasawa Shoun Co., Ltd.) and 81 g of rice bran by dry weight and mix well so that the water content is 65%. This was prepared, packed in a polypropylene 850 ml bottle (trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.), made a hole with a diameter of 1 cm from the center of the bottle mouth, and then plugged with a cap. . This was pasteurized at 120 ° C. for 60 minutes to prepare a culture medium. The culture medium was inoculated with solid inoculums of lyophilum ulmarium Lu1-172, Lu1-173, Lu1-174, and Lu1-181 strains, and cultured at 25 ° C. for 30 days to obtain cultured mycelia. The fruit body generating group was obtained by culturing for 40 days. After removing 1 cm of the upper mycelium layer of the fruiting body leaving the central part (fungal scraping), adding 20 ml of tap water to sufficiently absorb water, and removing excess water, the temperature is 15 ° C. and the humidity is 90% or more. Under the conditions, the Lu1-172 strain was cultured for 12 days, the Lu1-173 strain was cultured for 11 days, and the Lu1-174 and Lu1-181 strains were cultured for 10 days to form fruiting body primordium, irradiated with light, and further Lu1- The 172 strain was cultured for 14 days, the Lu1-173 strain for 13 days, the Lu1-174 strain for 11 days, and the Lu1-181 strain for 12 days to obtain fruiting bodies. The yields of the fruiting bodies were 181 g for the Lu1-172 strain, 189 g for the Lu1-173 strain, 173 g for the Lu1-174 strain, and 159 g for the Lu1-181 strain, which were high quality.
比較例８
リオフィラム ウルマリウムＫ−０２５９、ＩＦＯ９６３７、ＩＦＯ３０５２５株を使用する点と、菌掻き後に１５℃、湿度９０％以上の条件下でＫ−０２５９株とＩＦＯ９６３７株は１２日間、ＩＦＯ３０５２５株は１０日間培養して子実体原基を形成させ、光を照射して、更にＫ−０２５９株は１５日間、ＩＦＯ９６３７株は１４日間、ＩＦＯ３０５２５株は１２日間培養を続けて子実体を得る点を除いては実施例７と同様に行って、子実体を得た。得られた子実体の収量はＫ−０２５９株が１０７ｇ、ＩＦＯ９６３７株が１００ｇ、ＩＦＯ３０５２５株が１５３ｇで、品質は良好であった。ニンジン搾汁粕を用いた培養基ではＬｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１株の方がリオフィラム ウルマリウム Ｋ−０２５９、ＩＦＯ９６３７、ＩＦＯ３０５２５株より収量が優れていた。
Comparative Example 8
Liophyllum ulmarium K-0259, IFO9637, IFO30525 strains are used, and after sterilization, K-0259 strain and IFO9637 strain are cultured for 12 days and IFO30525 strain for 10 days under conditions of 15 ° C. and humidity of 90% or more. Exactly as in Example 7 except that the primordial primordium was formed and irradiated with light, and further cultured for 15 days for K-0259 strain, 14 days for IFO9637 strain, and 12 days for IFO30525 strain to obtain fruiting bodies. We went in the same way to get a fruiting body. The yield of fruit bodies obtained was 107 g for the K-0259 strain, 100 g for the IFO 9637 strain, and 153 g for the IFO 30525 strain, and the quality was good. In the culture medium using carrot juice extract, the yields of Lu1-172, Lu1-173, Lu1-174, and Lu1-181 were superior to those of Riofilum ulmarium K-0259, IFO9637, and IFO30525.
実施例８
ニンジン搾汁粕（ニンジン根茎の絞り粕）２７ｇ、針葉樹オガクズ〔（有）唐澤商運製〕２７ｇ、コーンコブ〔タカラ物産（株）製〕２７ｇ、米糠５４ｇ、マメカワ１８ｇ、フスマ１８ｇを乾燥重量で用いる点と、菌掻き後に１５℃、湿度９０％以上の条件下でＬｕ１−１７２株、Ｌｕ１−１７３株、及びＬｕ１−１７４株は１２日間、Ｌｕ１−１８１株は１０日間培養して子実体原基を形成させ、光を照射して、更にＬｕ１−１７２株とＬｕ１−１７４株は１３日間、Ｌｕ１−１７３株は１４日間、Ｌｕ１−１８１株は１２日間培養を続けて子実体を得る点を除いては実施例７と同様に行って、子実体を得た。得られた子実体の収量はＬｕ１−１７２株が１８２ｇ、Ｌｕ１−１７３株が１９４ｇ、Ｌｕ１−１７４株が１８３ｇ、Ｌｕ１−１８１株が１７９ｇであり、それぞれ高品質なものであった。
Example 8
Carrot squeezed rice cake (carrot rhizome squeezed rice cake) 27g, coniferous sawdust [manufactured by Karasawa Trading Co., Ltd.] 27g, corn cob [manufactured by Takara Bussan Co., Ltd.] 27g, rice bran 54g, mamekawa 18g, bran 18g in dry weight After the bacteria are scraped, the Lu1-172 strain, the Lu1-173 strain, and the Lu1-174 strain are cultured for 12 days and the Lu1-181 strain is cultured for 10 days under conditions of 15 ° C. and a humidity of 90% or more. After forming a group and irradiating with light, the Lu1-172 and Lu1-174 strains are cultured for 13 days, the Lu1-173 strain is cultured for 14 days, and the Lu1-181 strain is cultured for 12 days to obtain fruiting bodies. Except for this, the same procedure as in Example 7 was performed to obtain fruit bodies. The yields of the fruiting bodies obtained were 182 g for the Lu1-172 strain, 194 g for the Lu1-173 strain, 183 g for the Lu1-174 strain, and 179 g for the Lu1-181 strain, which were high quality.
比較例９
リオフィラム ウルマリウムＫ−０２５９、ＩＦＯ９６３７、ＩＦＯ３０５２５株を使用する点と、菌掻き後に１５℃、湿度９０％以上の条件下でＫ−０２５９株は１５日間、ＩＦＯ９６３７株は１２日間、ＩＦＯ３０５２５株は１１日間培養して子実体原基を形成させ、光を照射して、更にＫ−０２５９株は１５日間、ＩＦＯ９６３７株は１４日間、ＩＦＯ３０５２５株は１２日間培養を続けて子実体を得る点を除いては実施例８と同様に行って、子実体を得た。得られた子実体の収量はＫ−０２５９株が９５ｇ、ＩＦＯ９６３７株が１３６ｇ、ＩＦＯ３０５２５株が１７３ｇで、品質は良好であった。ニンジン搾汁粕を用いた培養基ではＬｕ１−１７２、Ｌｕ１−１７３、Ｌｕ１−１７４、及びＬｕ１−１８１株の方がリオフィラム ウルマリウム Ｋ−０２５９、ＩＦＯ９６３７、ＩＦＯ３０５２５株より収量が優れていた。
Comparative Example 9
Ryophyllum ulmarium K-0259, IFO9637, IFO30525 strain is used, and after sterilization, K-0259 strain is cultured for 15 days, IFO9637 strain is cultured for 12 days, and IFO30525 strain is cultured for 11 days under conditions of 15 ° C. and humidity of 90% or more. This was carried out except that fruit body primordia were formed and irradiated with light, and the K-0259 strain was cultured for 15 days, the IFO 9637 strain was cultured for 14 days, and the IFO 30525 strain was cultured for 12 days. In the same manner as in Example 8, fruit bodies were obtained. The yield of fruit bodies obtained was 95 g for the K-0259 strain, 136 g for the IFO 9637 strain, and 173 g for the IFO 30525 strain, and the quality was good. In the culture medium using carrot juice extract, the yields of Lu1-172, Lu1-173, Lu1-174, and Lu1-181 were superior to those of Riofilum ulmarium K-0259, IFO9637, and IFO30525.
実施例９
アシタバ搾汁粕、針葉樹オガクズ〔（有）唐澤商運製〕、コーンコブ〔タカラ物産（株）製〕、及び米糠を乾燥重量で表２の配合で計量し、よく混合して水分含有率６５％となるように調製し、これをポリプロピレン製８５０ｍｌビン〔商品名：ブロービンＳ−８５０、信越農材（株）製〕に詰めてビン口中央より下方に向かい直径１ｃｍの穴をあけた後、キャップで打栓した。これを１２０℃、６０分間高圧殺菌して培養基を調製した。該培養基にリオフィラム ウルマリウム菌株としてリオフィラム ウルマリウム Ｋ−０２５９株の固体種菌を接種し、２５℃の条件下で３０日培養して培養菌糸を得た後、更に５５日培養して子実体発生基を得た。該子実体発生基の上部菌糸層１ｃｍを中央部を残して除去し（菌掻き）、水道水２０ｍｌを加えて充分に吸水させた後、余剰の水を除いて１５℃、湿度９０％以上の条件下で１４日間培養して子実体原基を形成させ、光を照射して更に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表２の通りであり、それぞれ高品質なものであった。
Example 9
Ashitaba squeezed rice cake, coniferous sawdust (manufactured by Karasawa Trading Co., Ltd.), corn cob (manufactured by Takara Bussan Co., Ltd.), and rice bran were weighed according to the formulation shown in Table 2 in dry weight, mixed well, and a moisture content of 65 %, And this was packed in a polypropylene 850 ml bottle [trade name: Blowbin S-850, manufactured by Shin-Etsu Agricultural Materials Co., Ltd.] and a hole with a diameter of 1 cm was made downward from the center of the bottle mouth, It was stoppered with a cap. This was pasteurized at 120 ° C. for 60 minutes to prepare a culture medium. The culture medium was inoculated with a solid inoculum of Riofilum ulmarium K-0259 strain as the lyophilum ulmarium strain, cultured for 30 days at 25 ° C. to obtain a cultured mycelium, and further cultured for 55 days to obtain a fruiting body generating group. It was. After removing 1 cm of the upper mycelium layer of the fruiting body leaving the central part (fungal scraping) and adding 20 ml of tap water to sufficiently absorb water, the excess water is removed and the temperature is 15 ° C. and the humidity is 90% or more. The fruit body primordium was formed by culturing for 14 days under the conditions, and the fruit body was obtained by irradiating light and further culturing for 14 days. The yield of the fruiting bodies obtained was as shown in Table 2, and each was of high quality.
比較例１０
アシタバ搾汁滓１３０ｇのみを使用した点以外は、実施例９と同様に行ない、子実体を得た。得られた子実体の収量は４８ｇであり、アシタバ搾汁滓とセリ科植物由来の処理物からなる基材以外の混合比率が、１：０．２〜２０の範囲でアシタバ搾汁滓単独よりも良好であった。
Comparative Example 10
A fruit body was obtained in the same manner as in Example 9 except that only 130 g of Ashitaba squeezed rice cake was used. The yield of the obtained fruit body is 48g, and the mixing ratio other than the base material which consists of a processed product derived from an Ashitaba squeeze plant and an Aceraceae plant is 1: 0.2-20 from Ashitaba squeeze squeeze alone. Was also good.
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