JPH02117332A - Novel poria cocos and culture thereof - Google Patents

Abstract

PURPOSE:To provide a novel Poria cocos forming sclerotia in a culture under the artificial environment using no soil, rapid in their growth rate and capable or producing sclerotia of uniform quality irrespective of weather conditions. CONSTITUTION:A novel Poria cocos, a T-1 strain one (FERM No.102255) is inoculated into a wood classified as Pinaceae, and the resultant wood is put to culture under the artificial conditions at 25-34 deg.C and pH2.5-3.5. For the resultant Poria cocos, the sclerotia, which differ from those put to culture in the field, have no outer layer; therefore, when to be prepared as a raw material for crude drug, said Poria cocos is not be required for removing the outer layer. Also this Poria cocos is larger in shape compared to conventional ones and is white in the color of the sclerotia.

Description

[Detailed Description of the Invention] [Prior Art] Poria cocos is a fungus belonging to the family Arunococcosaceae, and its sclerotia are used as the crude drug "Hagirei", including Keishi Hagiregan, Gorebei, and Hagirein. It is included in many Chinese herbal medicine formulations and has been used since ancient times to treat various diseases associated with chest aeration, long illness, fright, palpitations, pain in the lower part of the body, cold fever, frustration, 0 scorch, dry tongue, and inconvenience when urinating. Ta.

Recently, in addition to diuretic effect, anti-gastric ulcer effect, blood thinning effect,
It has been recognized to have immunostimulatory effects.

This bukuriyou is a member of the Imperial family (Pos.
It is defined in the 11th edition of the Japanese Pharmacopoeia that it is a sclerotium from which most of the outer layer has been removed.

Its distribution is in East Asia such as China, Korea, and Japan, North America such as the United States, Canada, and Australia.

The parasitic range of the North American species is wide, and its properties are different from those from East Asia, and there is currently no proof that the species produced in these countries are the same species.

[Problem to be solved by the invention]

Bukuriyou is formed by attaching to roots 10 to 30 cm underground near the stump of a pine tree that has been felled for 3 to 5 years, and is searched for and dug out using a method called ``Furei Tsuki''. This is the common harvesting method. In China, some methods are being used to artificially form sclerotia by burying sclerotia and pine wood together in the soil. This Chinese cultivation method uses the sclerotium itself, which is used as an herbal medicine, as an inoculum source, so cultivation efficiency is low, and since it is placed in a natural environment, it is greatly affected by the weather conditions of the year, making production unstable. However, there are drawbacks such as the need for a large cultivation area.

Therefore, the inventors conducted a wide selection from the natural world, found a new plant that does not have the above-mentioned drawbacks, and completed the present invention.

[Means to solve the problem]

The novel Bukuriyo of the present invention forms sclerotia when cultivated in an artificial environment without using soil, and the sclerotia are white in color, do not form an outer layer, and grow larger in shape than conventional ones. It is a fast-paced bukuriyo.

A representative example of this is the T-1 strain.

The present invention involves inoculating a novel pine tree with wood of the genus Pinus,
A new method for cultivating Bukuriyo by burying the wood in the soil and cultivating it; and a new method for culturing Bukuriyo by inoculating wood from the genus Pinus, and culturing the wood at 25-34°C, p) (2.5-3.5 This includes a new method for cultivating Bukuriyo.

The details of this new method for selecting Bukuriyo from the natural world are as follows.

The present inventors investigated the natural habitat based on various documents and information and collected sclerotia from all over the country.

Wash away the mud adhering to the surface of the sclerotia, and when the surface is almost dry, remove a portion of the bacterial strain using a cork polarizer with a diameter of 8. After sterilizing the tissue directly below the outer layer with 80% ethyl alcohol, place it on a PDA (potato This was placed on a agar) medium.

As a result, 48 bacterial strains shown in Table 1 were isolated.

Although a considerable difference was observed in the growth rate of hyphae among these strains, the optimum growth temperature was 32'C, the optimum growth temperature was around p) 13, and the physiological properties were almost the same. Next, all of these strains were cultivated according to the above-mentioned Chinese method, and bacterial strain formation was observed in two strains, of which only one strain, Pc-24, conformed to the Japanese Pharmacopoeia.

(Margins below this page) Furthermore, strains having the ability to form sclerotia were cultivated over the years, large and superior sclerotia were selected, and pure isolation was repeated to obtain the novel sclerotium of the present invention.

The novel Bukuriyo of the present invention is characterized in that the color of the sclerotia is white and does not form an outer layer when cultivated in an artificial environment without using soil, and the growth rate is fast, and the sclerotia are formed without adding soil. It has the following. The Bukuriyo T-1 strain is a representative example.

This novel Bukuriyo T-1 strain has been deposited with the National Institute of Microbial Technology, Agency of Industrial Science and Technology as Fiber Science and Technology Research Institute No. 10255.

The properties of the sclerotia of this novel Pc-24 from a mycological point of view are as follows.

1. Color 2. Shape 3. Size 4. Optimum growth temperature. White Color Hemispherical to elliptical hemispherical length 10-20c+s 32°C Pc-24 Brown hemispherical to irregular spherical length 5-10cm 32°C 5. Growth Optimum pH around 36, growth rate 45.2 kg/month around 4.9 kg/n (7/month, no need for soil for growth) The major characteristics of the novel Bukuriyo T-1 strain of the present invention are While sclerotia grown in the field have a brown outer layer, the sclerotia of the present invention do not have this outer layer.This outer layer needs to be removed when preparing it as a crude drug raw material, but the powder method There is no need to remove sclerotia formed by.

In addition, since the sclerotia of Bukuriyo are formed in the soil in the natural environment, soil was thought to be essential for the formation of sclerotia of Bukuriyo, but the T-1 strain of the present invention has the test results shown in Table 2. As shown in the figure, it is clear that soil is not necessarily required.

The T-1 strain of the present invention is at 25-34°C, pH 2,5-3
．． It can be cultivated under 5 artificial environments. Since it can be cultivated in this artificial environment, it is not affected by weather conditions unlike conventional cultivation in a natural environment, and has the advantage of being able to obtain products of uniform quality. As is clear from Table 2 below, this T-1 strain is 8.4 to 9.6 times more effective in an artificial environment than Pc-24, and 1.
The growth rate is 7 times faster, and the growth rate is 1.6 to 1.7 times faster in the untreated area where soil is not used in the T-1 line under an artificial environment than in the case where soil is used. In other words, this T-1 line can be said to have an extremely excellent growth rate.

The above description has been made of cases in which soil is used and not in an artificial environment, but the novel pine wood can also be inoculated into pine wood and the wood is buried in the soil to form sclerotia.

The cultivation temperature range is 25-34°C and the pH range is 2.5-3.5. The humidity should be about 25%, and it grows well in soil that contains such moisture (it will form a clump if you hold it for half a minute, but it will collapse if you drop it on the ground). Bukuriyo does not require direct sunlight for cultivation, and on the contrary, if exposed to direct sunlight, it will easily age or die. However, sunlight is closely related to a 1 degree rise in soil temperature, and mycelium grows during It is used for temperature and humidity through the soil. As nutrients, it requires carbohydrates, nitrogen, inorganic salts, and growth factors. However, Bukuriyo can secrete enzymes to break down cellulose and other components in horsetail pine and red pine trees and absorb necessary nutrients.

In addition to the above-mentioned pines, black pines and other trees such as lacquer, oak, cedar, and mulberry can also be used for cultivating Bukuriyou. Sandy soil with good air circulation and good drainage is suitable as the soil used for cultivating Bukuriyo, as Bukuriyo constantly absorbs oxygen from the air and releases carbon dioxide during the growth process.

〔Effect of the invention〕

The present invention provides a novel Bukuriyo T-1 strain that does not form an outer layer on sclerotia, can grow in an artificial environment, and has a fast growth rate. Since this new Bukuriyo can be cultivated artificially, sclerotia of uniform quality can be obtained regardless of weather conditions, making it an extremely excellent raw material for crude drugs.

Next, the present invention will be described in detail using examples.

Example 1 Pine wood whose moisture content was adjusted with water (about 4 cm in diameter, 20 cm in length)
Make a drill hole in the center of the tube (cm), place it in a heat-resistant container, and sterilize it in an autoclave. After this pine wood is left to cool, mycelium of the T-1 strain cultured in a sawdust medium is embedded in the drill hole, and then a styrofoam plug is provided to prevent drying. The container containing the inoculated pine wood is placed in an incubator at 30±1°C, which is close to the optimum temperature for growth of the pine tree, and the mycelium is allowed to spread throughout the company. Further, it is cultured for 1 to 3 months under the same conditions to obtain sclerotia. The yield after 3 months of cultivation is 1 rrr of pine wood.
It was 135.5 kg per person. The results are shown in Table 2 below.

(Margins below this page) System Condition Treatment Period Pc-24 under artificial environment Akadama soil 3 months Kanuma soil
3 months humus 3 months no treatment 3 months natural environment 12 months 24 months T-1 artificial environment Akadama soil 3 months Kanuma soil 3 months humus 3 months no treatment 3 months natural environment 12 months 24 months Harvest amount (Kg/Po) ) 10.2 8.7 9.4 14.6 37.2 85.8 83.4 78.9 135.5 14.7 61.6 Example 2 In place of water when adjusting the moisture content in Example 1 Adjust the pH to 2.5-3.0 using phosphate buffer, and then lower the temperature to 25
Culture was carried out in the same manner as in Example 1 except that the temperature was adjusted to a range of -34°C. The results are shown in Table 3 below.

Claims (4)

[Claims] (1) Sclerotia are formed through cultivation in an artificial environment that does not use soil, and the sclerotia are white in color, do not form an outer layer, and are larger in shape and faster growing than conventional ones. Bukryo. (2) The novel Bukryo according to claim 1, wherein the Bukryo is T-1 strain. (3) A novel method for cultivating Prunus japonicum according to claim 1, which comprises inoculating wood of the genus Pinus with the novel Prunus japonicum according to claim 1, and cultivating the wood by burying the wood in the soil. (4) A novel method for cultivating Prunus japonicum according to claim 1, which comprises inoculating wood of the genus Pinus and culturing the wood at 25 to 34°C and pH 2.5 to 3.5.