JP7465463B2 - How to grow Poria columbine - Google Patents

Description

The present invention relates to a method for cultivating Poria cocos.

Wolfiporia cocos Ryvarden et Gilbertson, Wolfiporia extensa or Poria cocos is a mushroom of the Polyporaceae family. The sclerotium of Wolfiporia cocos is known as the herbal medicine "Bukuryo" which has diuretic and sedative effects. 99.8% of the Bukuryo used in Japan is produced in China. In recent years, pesticides such as insecticides and herbicides have been found in imported Bukuryo, raising safety concerns. In addition, the price of Bukuryo has been rising sharply in China due to the rapid population growth and growing health consciousness.

The method of cultivating Poria columbine widely used in China is non-sterile log cultivation, in which felled logs are inoculated with seed fungus outdoors and then cultivated. Non-sterile log cultivation is an extensive method that does not include a sterilization process. This means that problems such as the effects of weather and the occurrence of pests can easily occur, resulting in a low yield of Poria columbine.

On the other hand, sterilized log cultivation is a method in which felled logs are sterilized, inoculated with a seed fungus, and then cultivated indoors in a sterile environment. For example, Patent Document 1 discloses an artificial cultivation method for Poria cocos using red pine sawdust as a substrate. Patent Document 2 discloses an artificial cultivation method for Poria cocos, in which sliced pine logs are placed in a bag, and seed fungus is inoculated and cultivated under sterilized conditions to obtain sclerotia within the bag.

International Publication No. 2014/156759 JP 2000-092986 A

Patent Document 1 and Patent Document 2 propose using pine or cedar as the substrate and logs. Pine and cedar contain antibacterial substances. The antibacterial substances may inhibit the growth of mycelium and suppress the growth of sclerotia. As a result, the yield of Poria cocos will decrease.

The present invention has been made in view of the above-mentioned circumstances, and an object of the present invention is to provide a method for cultivating Poria cocos, which can increase the yield of Poria cocos.

The cultivation method of Poria cocos according to the first aspect of the present invention comprises the steps of:
A seed inoculation process in which a seed fungus of Japanese cedar is inoculated onto a cut cedar log that has been cut for more than four months;
A seed fungus cultivation step of cultivating the seed fungus of the Japanese cedar and forming hyphae on the Japanese cedar log;
including.

In this case, the cedar logs are
More than nine months have passed since felling.
This may also be the case.

In addition, in the cultivation method of Poria cocos according to the first aspect of the present invention,
The seed fungus inoculation step and the seed fungus culture step are further performed on the cedar log after harvesting the porgy formed in the seed fungus culture step.
This may also be the case.

In addition, in the cultivation method of Poria cocos according to the first aspect of the present invention,
a sclerotium inoculation step of inoculating the cedar log on which the mycelium has been formed with sclerotia of Pinus arbutifolia;
A sclerotium culturing step of culturing the sclerotium;
Further comprising:
This may also be the case.

In addition, in the cultivation method of Poria cocos according to the first aspect of the present invention,
The cedar log after harvesting the porphyra formed in the sclerotium culture step is further subjected to the seed fungus inoculation step, the seed fungus culture step, the sclerotium inoculation step and the sclerotium culture step.
This may also be the case.

In addition, in the cultivation method of Poria cocos according to the first aspect of the present invention,
The sclerotium inoculation step and the sclerotium culture step are further performed on the cedar log after harvesting the sclerotium formed in the sclerotium culture step.
This may also be the case.

In addition, in the cultivation method of Poria cocos according to the first aspect of the present invention,
After harvesting the sclerotia formed in the sclerotial culture step, the cedar log is sterilized, and then the sclerotial inoculation step and the sclerotial culture step are further carried out.
This may also be the case.

In addition, in the seed fungus inoculation step, the cedar log is sterilized and then inoculated with the seed fungus of Matsuhori.
This may also be the case.

In addition, in the cultivation method of Poria cocos according to the first aspect of the present invention,
sterilizing the cedar logs while the first bag for holding the cedar logs is placed in a second bag;
This may also be the case.

It can increase the yield of bupleurum.

FIG. 1 is a diagram showing a flowchart of a cultivation method for Poria cocos in accordance with the first embodiment. FIG. 11 is a flowchart showing a cultivation method for Poria cocos in accordance with a second embodiment.

The following describes an embodiment of the present invention. Note that the present invention is not limited to the following embodiment.

(Embodiment 1)
The cultivation method of Poria corymbos in this embodiment includes a seed fungus inoculation step (step S1) and a seed fungus cultivation step (step S2), as shown in Fig. 1. Any log usable for cultivation of Poria corymbos can be used as the log. Examples of the log include conifers, pine trees, Japanese cedar, Japanese cypress, red pine, larch, Abies sachalinensis, black pine, Japanese pine (Babishou), Japanese stone pine, Unnan pine, and Chinese cinnamon. Preferably, the log is Japanese cedar, which is abundant in thinned wood in Japan.

The logs are ones that have been felled for 4 months or more. The period since felling is not particularly limited as long as it is 4 months or more, but may be, for example, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months. Preferably, logs that have been felled for 9 months or more, particularly preferably 12 months or more, are used. The felled logs may be left, for example, outdoors and exposed to wind and rain, or may be stored indoors until use. Before the seed inoculation step, the logs may be soaked in water for a predetermined time, for example, 2 days. Taking into consideration the operability in the seed inoculation step, the logs may be cut to any size immediately after felling or immediately before the seed inoculation step.

In the seed inoculation process, the logs are inoculated with the seed of A. arbutifolia. The seed of A. arbutifolia may be collected from the wild or obtained from a preservation institution such as the American Type Culture Collection (ATCC) or the National Institute of Agricultural Sciences (NIAS) GeneBank. The seed of A. arbutifolia may be inoculated into the logs by a known method.

In the seed culture step, the seed fungus of Matsuhodo is cultured and mycelium is formed on the logs. The culture temperature is, for example, 20 to 32°C, more preferably 22 to 30°C, and particularly preferably 23°C. The humidity during culture is preferably 60% or higher. The seed fungus of Matsuhodo may be cultured under light, but is preferably cultured in darkness. The seed inoculation step may be performed by placing the logs in any container. When the logs are kept in a container, it is preferable to seal the container to prevent the intrusion of germs and pests.

By cultivating the seed fungus of Matsuhodo for a certain period of time, for example 20 days or more, the mycelium will grow abundantly on the surface of the log. When the mycelium adhere to each other, loses moisture inside, and the tissue becomes hard and dense, a sclerotium, or bukyō, is formed. The bukyō formed during the seed fungus cultivation process can be harvested, peeled, and dried.

As shown in Example 1 below, if the time that has passed since felling of the logs is short, the growth of mycelium is suppressed, and the longer the time that has passed since felling, the greater the yield of Poria cocos. In the Poria cocos cultivation method according to this embodiment, logs that have been felled for more than four months are used, so the yield of Poria cocos can be increased.

In the above cultivation method for Bukryo, the logs may be cedar. Pine is often used to cultivate Bukryo, but it is expected that it will be difficult to obtain a stable supply of pine for Bukryo cultivation within Japan. Cedar is abundantly available as thinned wood, so costs can be kept down compared to pine. Furthermore, making effective use of thinned wood can contribute to forestry.

In the method for cultivating Poria columbine according to this embodiment, a seed fungus inoculation step and a seed fungus cultivation step may be further performed on the logs after harvesting the Poria columbine formed in the seed fungus cultivation step. This allows logs that have been felled for more than four months to be effectively reused. Reusing logs is advantageous from the perspective of protecting forest resources.

(Embodiment 2)
Next, a description will be given of a second embodiment. In the cultivation method of Poria corymborne according to the present embodiment, in the seed fungus inoculation step, the logs are sterilized and then inoculated with seed fungus of Poria corymborne.

Before sterilizing the logs, they may be soaked in water for a specified time, for example, two days. Taking into account ease of handling during sterilization, the logs may be cut to any size immediately after felling or immediately before sterilization. Preferably, the logs are sterilized in any container. Depending on the capacity of the container, the size of the logs is appropriately adjusted, and one log may be split and placed in a container. Also, one or more logs may be placed in one container.

Sterilization conditions are set according to the type and amount of logs. Sterilization of logs is performed, for example, with high-pressure steam. Sterilization temperatures are 80 to 150°C, 85 to 140°C, 90 to 130°C, or 95 to 120°C. Sterilization may be performed at multiple temperatures, for example, by maintaining a first temperature of 90 to 110°C or 95 to 105°C for a predetermined period of time, followed by sterilization at a second temperature of 100 to 140°C or 110 to 130°C, which is higher than the first temperature. Sterilization times are, for example, 0.5 to 4 hours, 1 to 3 hours, or 1.5 to 2 hours. After sterilization, the logs are preferably cooled, for example, to room temperature. When sterilizing logs at high temperatures, a heat-resistant bag is preferable as a container for holding the logs.

Since pine and cedar trees immediately after felling contain a lot of sap (resin), when sterilized at high temperatures, the heat-resistant bag holding the logs may melt due to the heated sap. If the heat-resistant bag melts, there is the inconvenience of a decrease in the yield of Poria cocos due to the effects of pest infestation and the proliferation of bacteria. For this reason, when sterilizing at high temperatures, it is preferable to sterilize the logs while the heat-resistant bag holding the logs (first bag) is placed inside another heat-resistant bag (second bag).

After four months or more have passed since felling, the resin in the logs has flowed out and decreased, which helps prevent damage to the heat-resistant bags during sterilization. Furthermore, by placing the heat-resistant bag containing the logs in another heat-resistant bag and sterilizing the logs, damage to the outer heat-resistant bag can be avoided. This reduces the need to repeat sterilization, making it possible to cultivate Poria cocos in a sustainable and efficient manner. Furthermore, costs can be reduced by reusing the outer heat-resistant bag as the inner heat-resistant bag.

In the seed inoculation step of this embodiment, sterilized logs are inoculated with pineapple seed. In this case, it is preferable to inoculate the logs with pineapple seed in a sterile facility such as a clean bench. During seed cultivation, it is preferable to seal the heat-resistant bag to prevent the intrusion of germs and pests. If the heat-resistant bag containing the logs is placed in another heat-resistant bag to sterilize the logs, it is preferable to inoculate the logs with pineapple seed in the seed inoculation step while maintaining that state.

When the seed fungus inoculation step and seed fungus cultivation step are further performed on the logs after harvesting the budding stalks formed in the seed fungus cultivation step, the logs may be sterilized in the seed fungus inoculation step and then inoculated with the budding stalks. By sterilizing the logs, it is possible to remove unwanted bacteria from the logs to be reused and increase the yield of budding stalks.

(Embodiment 3)
As shown in Fig. 2, the cultivation method of Poria columbine according to this embodiment includes a sclerotium inoculation step (step S3) and a sclerotium culture step (step S4) in addition to the seed fungus inoculation step (step S1) and seed fungus culture step (step S2) described in the above-mentioned embodiment 1 or embodiment 2. The seed fungus inoculation step and seed fungus culture step in the cultivation method of Poria columbine according to this embodiment are the same as those in the above-mentioned embodiment 1 or embodiment 2. The sclerotium inoculation step and the sclerotium culture step will be mainly described below.

In the sclerotium inoculation process, sclerotia of Matsuhodo (also called B. sclerotium) are inoculated onto the logs on which mycelium has formed. In inoculation of sclerotia, the sclerotia may be placed on the logs on which mycelium has formed in the seed fungus cultivation process. The amount of sclerotia is not particularly limited, but is, for example, 5 to 40 g or 10 to 30 g. The sclerotia may be immature or mature. The sclerotia may have the skin attached, or the skin may be removed. One or more sclerotia may be inoculated onto one log.

In the sclerotium culturing step, the sclerotia are cultured. The temperature during culturing the sclerotia is 20 to 35°C, 21 to 30°C, or 22 to 25°C. The humidity during culturing the sclerotia is preferably 60% or higher. The sclerotia of Pinus arbutifolia may be cultured under light, but is preferably cultured under darkness. During culturing of the sclerotia, it is preferable to seal the container that holds the logs to prevent the intrusion of germs and pests. The sclerotia are enlarged by culturing under such conditions for, for example, 60 to 120 days, preferably 80 to 100 days. The sclerotia may be cultured for, for example, 100 days or more, or for more than six months.

As shown in Example 2 below, the yield of Poria cocos increases the longer the time since felling. The cultivation method according to this embodiment can further increase the yield compared to the above-mentioned embodiment 1.

In addition, in the cultivation method of Poria columbine according to this embodiment, the log after the Poria columbine formed in the sclerotium cultivation step may be used for further steps of inoculation of the seed fungus, cultivation of the seed fungus, cultivation of the sclerotium, and cultivation of the sclerotium. If there are few mycelia on the surface of the log after the Poria columbine has been harvested, the log may be reused for the inoculation of the seed fungus step to promote the formation of mycelia on the log, which makes it easier to increase the yield of Poria columbine. In this case, if germs have developed on the surface of the log, the log may be sterilized in the inoculation of the seed fungus step and then inoculated with the seed fungus of Pinus arbutus. When the log is reused for the inoculation of the seed fungus step, it is preferable to use the log after the Poria columbine has been harvested without soaking it in water.

The sclerotium inoculation step and sclerotium cultivation step may be further carried out on the logs after the porgy leaves formed in the sclerotium cultivation step are harvested. In particular, if mycelium is widespread on the surface of the logs after the porgy leaves are harvested and no unwanted bacteria have developed, the logs may be used as is for the sclerotium inoculation step. On the other hand, if unwanted bacteria have developed on the surface of the logs, the logs to be reused may be sterilized and then inoculated with sclerotia of Matsuhodo porgy.

In another embodiment, a substrate for cultivating Bukryo is provided. The substrate for cultivating Bukryo comprises a log that has been cut down for at least four months. The size of the log is not particularly limited and is adjusted as appropriate. In another embodiment, a Bukryo cultivation kit is provided that comprises the substrate for cultivating Bukryo and a container. The container is not particularly limited, but is preferably a container made of a heat-resistant material. The container is, for example, a heat-resistant bag. In the Bukryo cultivation kit, the substrate for cultivating Bukryo and the container may be sterilized. Preferably, the substrate for cultivating Bukryo is held in the container and maintained in a sterilized state. In this way, the user can immediately perform the seed inoculation process on the substrate for cultivating Bukryo, thereby efficiently cultivating Bukryo.

The present invention will be explained in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1
The felled cedar logs were exposed to the elements for 0-2 months, 2-4 months, 4-6 months, 6-8 months, or 10-12 months after felling. The cedar logs were cut to a length that could fit into a heat-resistant bag with a filter (50 μm thick, 40φ filter hole diameter, 325 mm folded diameter x 62 mm gusset x 450 mm length; ST50-TMS 40φ, manufactured by F-Tech Co., Ltd.). The heat-resistant bag containing the cut cedar logs was sterilized with high-pressure steam. For sterilization, the temperature was raised to 100°C over 20 minutes, then to 118°C over 20 minutes, and maintained for 1 hour and 30 minutes. After sterilization, the heat-resistant bag was left to stand overnight in a room where the temperature could be controlled to cool the heat-resistant bag. If there was a risk of holes being made in the heat-resistant bag due to the sap becoming hot during sterilization, the heat-resistant bag containing the cedar logs was further placed in a heat-resistant bag for sterilization.

Under aseptic conditions, approximately 30 ml (20 g) of Matsuhodo seed fungus (TMIC-36031 strain) cultured on sawdust was inoculated onto the cedar logs in each heat-resistant bag. The bags were sealed with filters and cultured at 23°C in the dark. Five months after inoculation, sclerotia were harvested.

Example 2
After sterilizing the cedar logs in the same manner as in Example 1, the seed fungus of Matsuhodo was inoculated onto the cedar logs and cultured. After about one month, 20 g (fresh weight) of sclerotia of Matsuhodo were placed on the cedar logs. After four months, the sclerotia were harvested.

(result)
Heat-resistant bags containing cedar logs that were 0-2 months, 2-4 months, or 4-6 months old after felling had holes in the bottom due to the high temperature of the sap during sterilization. For cedar logs that were 0-2 months old after felling, holes were developed in almost all of the heat-resistant bags, but the percentage of heat-resistant bags with holes tended to decrease as the time since felling increased. Even when heat-resistant bags containing cedar logs were placed in another heat-resistant bag and sterilized, holes were developed in the outer heat-resistant bag for cedar logs that were not long after felling, but almost no holes were developed in the heat-resistant bags for cedar logs that were 6 months or more old after felling.

The period of exposure to wind and rain after felling is important in reducing damage caused by holes in the heat-resistant bag due to leakage of sap during high-temperature sterilization. It has been revealed that if the cedar logs are at least four months old after felling, placing the cedar logs in a heat-resistant bag inside another heat-resistant bag can reduce damage caused by holes in the heat-resistant bag due to sterilization.

The yield of sclerotia for each period after felling of cedar logs is shown in Table 1. Note that the "number of heat-resistant bags" in Table 1 was counted as one even if a heat-resistant bag containing a cedar log was placed inside another heat-resistant bag.

In Example 1, when cedar logs that had been felled for 0 to 2 months were used, the growth of the inoculated Matsuhodo seed fungus was inhibited, mycelium did not spread on the cedar logs, and sclerotia formation was hardly observed. The yield of sclerotia increased as the time since felling increased.

In Example 2, the yield was higher overall than in Example 1, and the yield increased the longer the time since felling. An average yield of 267.9 g was obtained from 10-12 month-old cedar logs. This shows that the time that cedar logs have been in operation since felling is important for the spread of mycelium and the subsequent formation of sclerotia. Because cedar contains antibacterial substances, it is believed that if the seed fungus is inoculated immediately after felling, mycelium elongation is inhibited and sclerotia will not grow or growth will be significantly delayed.

The present invention allows for various embodiments and modifications without departing from the broad spirit and scope of the present invention. Furthermore, the above-described embodiments are intended to explain the present invention and do not limit the scope of the present invention. In other words, the scope of the present invention is indicated by the claims, not the embodiments. Furthermore, various modifications made within the scope of the claims and within the scope of the meaning of the invention equivalent thereto are considered to be within the scope of the present invention.

The present invention is suitable for the production of Poria cocos.

Claims (9)

A seed inoculation process in which a seed fungus of Japanese cedar is inoculated onto a cut cedar log that has been cut for more than four months;
A seed fungus cultivation step of cultivating the seed fungus of the Japanese cedar and forming hyphae on the Japanese cedar log;
Including, how to grow Poria columbine. The cedar logs are
More than nine months have passed since felling.
A method for cultivating Poria cocos according to claim 1. The seed fungus inoculation step and the seed fungus culture step are further performed on the cedar log after harvesting the porgy formed in the seed fungus culture step.
A method for cultivating Poria cocos according to claim 1 or 2. a sclerotium inoculation step of inoculating the cedar log on which the mycelium has been formed with sclerotia of Pinus arbutifolia;
A sclerotium culturing step of culturing the sclerotium;
The method for cultivating Poria cocos according to any one of claims 1 to 3, further comprising: The cedar log after harvesting the porphyra formed in the sclerotium culture step is further subjected to the seed fungus inoculation step, the seed fungus culture step, the sclerotium inoculation step and the sclerotium culture step.
The method for cultivating Poria cocos according to claim 4. The sclerotium inoculation step and the sclerotium culture step are further performed on the cedar log after harvesting the sclerotium formed in the sclerotium culture step.
The method for cultivating Poria cocos according to claim 4 or 5. After harvesting the sclerotia formed in the sclerotial culture step, the cedar log is sterilized, and then the sclerotial inoculation step and the sclerotial culture step are further carried out.
The method for cultivating Poria cocos according to claim 6. In the seed fungus inoculation step, the cedar log is sterilized and then inoculated with the seed fungus of the Japanese holly.
A method for cultivating Poria cocos according to any one of claims 1 to 7. sterilizing the cedar logs while the first bag for holding the cedar logs is placed in a second bag;
The method for cultivating Poria cocos according to claim 7 or 8.

Images