JP4230309B2 - How to grow Hanabiratake - Google Patents

Description

  TECHNICAL FIELD The present invention relates to a method for cultivating a bamboo shoot (Sparasiss crispa), and more particularly to a method for cultivating a bamboo shoot using a liquid inoculum.

  In the current cultivation of edible mushrooms, in the process, by performing empirically obtained generation operations such as low-temperature stimulation, fungus scraping, water injection, cover soil, etc. Yes. By performing this operation, efficient production becomes possible.

  By the way, all the mushrooms currently cultivated on a commercial scale are mushrooms with high lignin resolution called white rot fungi. On the other hand, mushrooms with low lignin resolution called brown rot fungi were not cultivated on a commercial scale. As one of the causes, it is conceivable that it is difficult to promote the generation of the child entity by the generation operation and to control the formation time of the child entity.

  Hanabiratake, which is classified as a brown rot fungus, is an extremely edible mushroom that grows in larch, is chewy, has a chewy white color, and is characterized by its shape like leaf peony. Although this bamboo shoot has not been able to form fruit bodies in an artificial medium for a long time, a new cultivation method that can be cultivated in a relatively short period of time has recently been established (for example, see Patent Document 1), and supply on a commercial scale Became possible. The use in the field of pharmaceuticals and foods has been proposed for an extract containing β-glucan as a main component obtained by extraction from this flower (for example, see Patent Documents 2 and 3).

  However, conventional Hanabira bamboo cultivation has a problem that the fruiting body formation rate at the time of cultivation is extremely low and the generation time varies. This problem has been difficult to solve even if the generation operation usually used for white rot fungi is performed. As a means for solving this problem, it has recently been reported that the fruit body formation rate of the alga is greatly improved by using a method of inoculating a different mushroom inoculum for the fruit body generation operation (see Patent Document 4). ). However, when this method is used, there is a problem that the cost increases due to an increase in the number of processes.

  On the other hand, a method using a liquid inoculum in cultivation of mushrooms is known. For example, in a method in which a continuously liquid-cultured fungus is used as a seed (see Patent Document 5), the hypha is dispersed and the viability of the mycelium is increased by continuous culture, and the spread rate of the mycelium in the solid medium is increased. It has been shown that it is possible. In addition, a method for artificial cultivation of mushrooms has been proposed in which a completely sterilized medium can be used by mixing a liquid-cultured mushroom cell body and a nutrient-containing culture solution into a sterilized solid medium to form a fungus bed ( (See Patent Document 6). Furthermore, it is disclosed that the culture period can be shortened by radiating and inoculating the liquid inoculum in the inoculation device in the inoculation hole opened in the medium and radiating and cultivating the liquid inoculum to substantially the entire inner surface of the inoculation hole and the medium surface of the bottle mouth (patent) Reference 7).

However, in the conventional cultivation method using liquid inoculum, there is no description that the operation of generating fruit bodies can be omitted, and no example of application to the artificial cultivation of hanabiratake has been reported.
JP-A-11-56908 JP 2000-217543 A JP 2002-125460 A JP 2002-369621 A JP-A-5-192036 JP-A-6-62662 JP 2003-158921 A

  It is an object of the present invention to provide a cultivation method more suitable for commercial cultivation without increasing the cost by increasing the fruiting body formation rate without performing a generation operation for Hanabiratake which has been recognized as useful. To do.

  As a result of intensive studies to solve the above problems, the present inventors have determined that the inoculum is not a mycelium cultured in a commonly used sawdust medium, but a liquid-cultured mycelium, which is uniformly and densely formed on a medium substrate. It was found that by increasing the mycelial density by inoculating the cell body, it is possible to increase the fruiting body formation rate without performing the generation operation, and the present invention has been completed.

That is, the present invention relates to a method for cultivating agaric bamboo, which is characterized in that a mycelium obtained by liquid culture is inoculated into a solid medium as an inoculum and grown without performing fruiting body generation operation. In addition, in the method for cultivating Hanabiratake, the method for cultivating Hanabiratake is characterized by cultivating mycelium obtained by liquid culture and then inoculating a solid medium as an inoculum. It is.

  According to the cultivation method of the present invention, it is possible to increase the fruiting body formation rate without performing the generation operation, thereby shortening the cultivation days of the bamboo flakes and greatly increasing the cultivation efficiency such as simplifying the cultivation process. In addition, a higher yield can be obtained.

  The method for cultivating Hanabira teke of the present invention can be divided into two steps: a step of preparing an inoculum by liquid culture, and a step of inoculating and cultivating the obtained inoculum on a solid medium.

  First, the inoculum preparation process by liquid culture is demonstrated.

  This process consists of the following operations. 1) Inoculate the stock strain on a plate medium. The medium material may be a material used for normal bacterial culture such as PDA medium. 2) This is cultured at around 25 ° C. and grown to a sufficient size, and then the medium is cut into small pieces having a diameter of 5 to 6 mm. The culture period at this time usually takes 2 weeks or more. 3) Inoculate the sterilized liquid medium with the cut medium. At this time, a plurality of small pieces may be used. 4) Perform liquid culture and use it as an inoculum when mycelia grow on the entire liquid medium. 5) For the inoculum, a mycelium cultured in liquid culture may be used after being transferred to a liquid medium.

  Hanabiratake used in the present invention is not particularly limited to any strain, and any strain can be used. As a method for obtaining the garlic mushroom, one that is grown in the wild may be collected, or a conserved strain of the Japan Mushroom Research Institute may be ordered. Furthermore, if a small piece of garlic mushroom commercially available for raw consumption is cut as aseptically as possible, then inoculated and cultured in a PDA agar medium, and then transferred to a slant, a garlic mushroom for storage can be obtained.

  It is not necessary to use special media components for liquid culture used in the present invention. Specifically, an inorganic or organic nitrogen source can be used as the nitrogen source of the medium component, but an organic nitrogen source is preferably used from the viewpoint of growth rate. As the carbon source, a commonly used carbon source such as a monosaccharide such as glucose or a polysaccharide such as starch can be used. Moreover, adding growth factors such as trace elements and vitamins as needed is no different from the usual case. The pH is 2.5 to 8.0, preferably 3.0 to 7.0, and most preferably 3.5 to 5.5. It is preferable to add an insoluble component to the medium because it can grow uniformly. As the insoluble component, soy flour or the like is used, and these may be added in an amount of about 0.05% to several percent with respect to the volume of the liquid medium.

  As the culture conditions for liquid culture, the culture temperature is 15 ° C. to 30 ° C., preferably 18 ° C. to 28 ° C., and most preferably 20 ° C. to 25 ° C. Any method such as shaking culture may be used. The culture time may be taken as a standard for ending the culture until the mycelium concentration is 2 mg / ml or more, and it varies depending on the strain, but it is several days to several weeks, and usually 2 to 3 weeks.

  Next, the process of inoculating and cultivating the liquid inoculum in a solid medium will be described.

  This process consists of the following operations. 1) Prepare a solid medium in the cultivation bottle. It is desirable to have an inoculation hole in the solid medium. 2) Inoculate the solid medium with the liquid inoculum obtained in the previous step. 3) Culture the inoculated medium under normal conditions. For those in which fruiting body formation was observed, the lids of the cultivation bottles were sequentially removed and transferred to the cultivation room set under normal conditions to grow the fruiting bodies.

The solid medium is preferably a sawdust medium. Larch sawdust is used as a medium base material, and flour is mixed with 1% to 30%. The flour is preferably 3% to 20%, more preferably 5% to 10%. As the medium substrate, corn cob, a mixture of hardwood and conifer can be used. About 520 g of this is normally filled into an 850 cc polypropylene cultivation bottle. At this time, it is preferable to make an inoculation hole in the center of the medium because the amount of inoculation can be increased, so that the mycelia can easily rotate around the entire medium and the ventilation is improved. Opening the inoculation hole results in advantages such as shortening the time to harvest and increasing the yield. The size of the inoculation hole is determined according to the instrument used for inoculation, but it is sufficient that the cross-sectional area is approximately 7 mm ² or more. Moreover, it is better to open a plurality of inoculation holes. The solid medium is sterilized by a method such as high pressure sterilization.

  In the present invention, it is preferable to aseptically pulverize before inoculating the liquid cultured inoculum. Since the mycelium cultured in liquid is usually in the form of slurry or diatom, the mycelium can be uniformly inoculated into the inoculation hole and the upper surface of the medium by inoculating after pulverization. A high-density fruiting body can be obtained. The pulverization can be performed using a homogenizer, and it is sufficient if the slurry-like or algal-like mycelium disintegrates into a uniform dispersed state.

  Inoculation of the inoculum may be performed aseptically using an injector, or may be performed using a commercially available liquid inoculator. At this time, it is preferable to uniformly inoculate the mycelium on the inoculation hole and the upper surface of the medium. The optimal inoculation amount per cultivation bottle is 5 to 30 mL in terms of liquid volume, 7 to 25 mL is even better, and 10 to 20 mL is most preferable. A higher mycelium concentration is better. It should be 0.5 mg / ml or more, preferably 1 mg / ml or more, more preferably 2 mg / ml or more.

  As culture conditions for the medium inoculated with the inoculum, the temperature is most preferably 15 ° C. to 30 ° C., preferably 18 ° C. to 28 ° C., 20 to 25 ° C., and the humidity is 50% to 80%, preferably 55% to 75%. 60% to 70% is most preferable. By culturing by the method as described above, fruit body formation of approximately 90% or more can be seen without any special operation. For those in which fruiting body formation was observed, the bottles of the cultivation bottle were sequentially removed, and the normal conditions, that is, the temperature of 10 ° C. to 30 ° C., preferably 15 ° C. to 28 ° C., most preferably 17 to 25 ° C., humidity 80 % Or more, preferably 85% or more, and most preferably 90% or more. By such cultivation, fruit bodies can be stably harvested 130 g or more (per cultivation bottle).

Example 1
Hanabiratake stock strain (slant preserved from unitika commercial agaricus mushroom fragment) was inoculated on PDA plate medium and cultured at 25 ° C. for 3 weeks. The medium was cut into small pieces having a diameter of 5 to 6 mm, and inoculated into a liquid medium (Kobo extract 0.5%, glucose 2%, pH 4.0) previously sterilized. After static culture at 25 ° C., the cells were transferred to a liquid medium after 2 weeks. At the time of inoculation, the mycelium was aseptically crushed at 10,000 rpm for 1 minute using a homogenizer together with the medium and inoculated into the liquid medium. The inoculum is 10 mL (100 mg dry cell weight) per 100 mL of medium. This was further cultured for 2 weeks, and mycelia grown on the entire liquid medium were used as seeds. The concentration of the inoculum used was 3.5 mg / mL. At the time of inoculation, this inoculum was aseptically pulverized at 10,000 rpm for 1 minute using a homogenizer and inoculated into sawdust medium.

  As a medium substrate for the sawdust medium, larch sawdust was used, 7% of flour was mixed therein, and water was added to adjust the water content to 61%. This was filled into 850 cc polypropylene cultivation bottles, and an inoculation hole with a diameter of 20 mm was made in the center of the cultivation bottle using a commercially available punching machine, and then pasteurized at 110 ° C. for 3 hours. Thereafter, the inoculum was inoculated after the medium temperature dropped to 30 ° C. or lower. The inoculation amount is 6 mL (dry cell weight 21 mg) in the inoculation hole, and 4 mL (dry cell weight 14 mg) on the top of the medium. When this was cultured at 23 ° C. and 65% humidity, almost 100% of fruiting bodies were formed after 42 days. About what the fruit body formation was seen, the lid | cover of the cultivation bottle was removed sequentially, it moved to the cultivation room of temperature 19 degreeC and 90% of humidity, and the fruit body was grown. When cultivated with 160 bottles, the average yield per bottle was about 150 g.

Example 2
Seven strains NS100, NS422, NS423, NS424, NS425, TMC504, and TMC572, which are conserved strains of Hanabiratake obtained from Japan Mushroom Research Institute, were inoculated and cultivated in the same manner as in Example 1, and all fruit bodies The formation of was seen. Table 1 shows the results for each strain.

比較例１
種菌としてオガクズ培地に生育させた菌糸体を用いた以外は実施例１と全く同様の操作により接種を行い培養した。

Comparative Example 1
The seeds were inoculated and cultured in exactly the same manner as in Example 1 except that mycelia grown on sawdust medium were used.

About 56% of fruiting bodies were observed after 56 days. However, there was considerable variation in the timing of formation. About what the fruit body formation was seen, the lid | cover of the cultivation bottle was removed sequentially, it moved to the cultivation room of temperature 19 degreeC and 90% of humidity, and the fruit body was grown. The final fruiting body rate was about 20%, and the average yield per 160 was about 90 g.
Comparative Example 2
The same culture as in Comparative Example 1 was performed, and when the formation of fruiting bodies was observed at about 15%, the surface of the medium was scraped off for all the fruiting bodies that were not seen, and the different types of mushrooms were found. Of inoculum. As the heterologous mushrooms, 2 g of inoculum prepared by cultivating spinach mushrooms in the same sawdust medium was used.

  In about 2 weeks after the inoculation, the formation of the fruit body of Hanabiratake was observed, and the formation rate was about 90%. About what the fruit body formation was seen, the lid | cover of the cultivation bottle was removed sequentially, it moved to the cultivation room of temperature 19 degreeC and 90% of humidity, and the fruit body was grown. The average yield per bottle per 160 cultivation bottles was about 130 g.

  Table 2 summarizes the contents in which significant differences were found between Example 1 and Comparative Examples 1 and 2. It can be seen that the period until the fruit body formation is shortened by inoculating the liquid. It can also be seen that the fruiting body formation rate has increased to almost 100%, the formation time is well aligned, and the average yield has also increased.

このように、液体種菌を用いることによって、子実体発生操作が不要となり、子実体形成率が上昇し、その形成までの時期が短縮できるなどハナビラタケの栽培効率を大幅に上げることができ、さらには収量も上げることができる。これらより商業規模での栽培において大きくコストダウンを図ることが可能となる。

In this way, by using the liquid inoculum, the fruiting body generation operation becomes unnecessary, the fruiting body formation rate increases, the time to formation can be shortened, etc. Yield can also be increased. From these, it becomes possible to greatly reduce the cost in cultivation on a commercial scale.

Claims (2)

A method for cultivating agaric bamboo, which comprises inoculating a mycelium obtained by liquid culture into a solid medium as an inoculum and allowing it to grow without performing fruiting body generation operation . 2. A method for cultivating garlic mushrooms according to claim 1, wherein the mycelium obtained by liquid culture is pulverized and then inoculated into a solid medium as an inoculum.