KR20000073245A - Artificial raising method of the entomogenous fungi by means of the mutant silkworm, Nd, as a host - Google Patents

Abstract

PURPOSE: An artificial cultivation method of Cordyceps using Nd mutant silkworm instead of silkworm is provided to increase the output with reduced labor and improve the fruit body forming rate. CONSTITUTION: The method comprises the following steps: (i) diluting a spore of Paecilomyces japonica to 107-108 spores/ml; (ii) inoculating the diluted spore to a host silkworm in the larval stage which cast off the skin 4times by spraying; (iii) leaving the host at 26-30°C under 95% of relative humidity for 12hr and breeding with mulberry leaves; and then (iv) if infections is confirmed, forming a fruit body by protecting the host at 21-25°C under 95% of relative humidity. The first hybrid of Nd mutant silkworm is used for the host.

Description

Artificial raising method of the entomogenous fungi by means of the mutant silkworm, Nd, as a host}

The present invention is a method for artificially cultivating Cordyceps sinensis using gajam, instead of gajam ( By using a mutant silkworm as a host insect, the conventional manual process of cutting the cocoon by selecting a cocoon can be omitted, thereby reducing labor and improving the formation rate of fruiting bodies.

Cordyceps sinensis (冬蟲夏草) is a name given to mean insects in winter and mushrooms in summer, and it is an insect parasitic mushroom that occurs when insects enter the insect body. This mushroom has long been known as a elixir of immortality. In recent years, modern scientific analytical techniques have confirmed pharmacological effects such as anticancer, immune enhancing and anti-fatigue effects in Cordyceps. .

However, wild cordyceps was extremely low in density, so it was difficult to collect it in the field and secure the quantity. Therefore, attempts were made to artificially cultivate it and supply it in large quantities. As a result, it is a gossip insect as Paecilomyces japonica; Patent Publication No. 98-23907 (98. 7. 6)), J300 Paecilomyces sp. J300; Patent Publication No. 98-23908 (98. 7. 6)), pupa rod-shaped Snow Cordyceps (Paecilomyces farinosa; Patent Publication No. 98-25197 (July 6, 1998)) and pupa Cordyceps militaris (Patent No. 98-25198 (July 6, 1998)) have been developed and patented. have.

The above method is inoculated spores of Cordyceps fungus into larvae or chrysalis of Gaza, breed them according to the commercial law, put them on the ((upper work) to make cocoons, and then cut the cocoons after a certain period of time to take out the pupa. The pupa (cured pupa) infected with Cordyceps fungus was selected and then protected under moderate temperature and humidity conditions to artificially cultivate Cordyceps fungus.

However, the work of raising silkworms on the top of the silkworms (manning work), constructing the cocoons, and harvesting them from the rice paddies (harvesting) was an essential process in the conventional sheep farming farms that produce cocoons. When cultivating Cordyceps sinensis, cocoons are not only an obstacle to confirming the infection of Cordyceps fungi, but also an unnecessary entity that interferes with the growth of fruiting bodies in the chrysalis. However, since silkworms repair cocoons are the same basic physiological actions as human excretion, they are not only difficult to artificially prevent the silkworm's earth and sand, but also to mobilize diarrhea. Silkworm will not grow normally and die. This is because the cocoon has an ecological meaning of protecting itself from the external during the pupa period when it is impossible to move, and at the same time, it contains the physiological meaning of excretion of silk material, a surplus protein accumulated in the silk gland, in the form of cocoon. However, since there has not yet been developed a simple way to prevent silkworms from producing cocoons, the cocoons harvested through foot and shoulder work during artificial cultivation of cordyceps are forced to double the unnecessary work of removing a lot of labor. It is being done.

If silkworms don't make cocoons, you can skip the process of stepping and veterinary work, as well as the growth of fruiting bodies in the chrysalis. By the way, Na Yong (which has the gene Nd Mutant silkworms do not have cocoons from birth. This is because silkworms do not accumulate silkworms in the body, so it is not accompanied by physiological desire to excrete excess protein. In view of this, the present inventors Using a hybrid hybrid of Nd mutant silkworms and Nd mutant silkworms, which are one of the mutant silkworms, as a cultivation host of Cordyceps sinensis, a method was found to reduce labor and increase the fruiting body formation rate of Cordyceps sinensis cultivation and completed the present invention Was done.

Accordingly, an object of the present invention is to provide a method that can reduce the labor force, and also increase the rate of formation of fruiting bodies by omitting the foot work, sowing work, and cutting work in artificial cultivation of Cordyceps sinensis.

Figure 1 shows the dilution of spores of Paecilomyces japonica sprayed on 5-year-old silkworm larvae and Cordyceps of Nd mutant silkworms and white jade according to time protected in high temperature and high humidity (28.5 ℃, 95% relative humidity) environment after inoculation. This is a comparison of infection rates.

Fig. 2 shows the fruiting bodies of Nd mutant silkworms and white jade according to the spore dilution concentration of Paecilomyces japonica sprayed on 5-year-old silkworm larvae and protected in high temperature and high humidity (28.5 ℃, 95% relative humidity) environment after inoculation. This is a comparison of rates.

Figure 3 shows the inoculation of 10 ⁸ spores / ml of Paecilomyces japonica inoculated with 1 Nd mutant silkworm hybrids when protected for 12 hours or 24 hours in a high temperature and high humidity (28.5 ℃, 95% relative humidity) environment This figure compares the infection rate of Cordyceps fungus in Baekjajam.

Figure 4 shows the inoculation of 10 ⁸ spores / ml of Paecilomyces japonica inoculated with 1 Nd mutant silkworm 1 hybrid when protected for 12 hours or 24 hours in a high temperature and high humidity (28.5 ℃, 95% relative humidity) environment This figure compares the fruiting body formation rate of Baekjajam.

Figure 5 is inoculated with 10 ⁸ spores / ㎖ dilution of Paecilomyces japonica, one hybrid Nd mutant silkworms were protected for 12 hours in a high temperature and high humidity (28.5 ℃, 95% relative humidity) environment, the general hybrid (white jade) Zam) compares the labor force and Cordyceps yields between the two plants when they are protected for 24 hours.

In order to achieve the above object, the method adopted in the present invention is characterized in that Nd mutant silkworms are used as host insects in artificial cultivation of Cordyceps sinensis using silkworms as host insects.

In addition, the method of the present invention is characterized by using one hybrid of a Nd mutant silkworm and a Japanese or Chinese species as a host insect.

Other objects and functions of the present invention will become apparent to those skilled in the art from the following detailed description of the invention.

Hereinafter, the present invention will be described in detail.

There are five strains known to have different characteristics in terms of genotyping, solubility, weight, and hatching ratios. Comparative investigation.

Unavailability, weight and hatching rate of silkworm silkworm Gongsinue ¹ Dissolution Rate ² (%) Soluble rate ³ (%) Weight (g) Spawning water Incubation rate (%) Nd 95.1 99.3 1.4 376 91.2 Nd ^H 93.6 25.5 1.2 407 83.3 p ³ Nd 92.2 26.1 1.0 351 74.5 Nd-s 72.1 50.4 1.2 367 62.8 Nd-t 82.5 3.8 1.2 - - White jade 93.2 0.0 1.7 434 92.7 Announcement: 150 heads per system 2. Dissolution Rate: Head count / Head count 3. Unavailability rate: Long head / long head

From Table 1, the solubilization rate, which is an indicator of the health of silkworms, was the highest in Nd (95.1%) and the solubilization rate was 99.3%. Nd was the heaviest in Nayong system, but did not reach Baekjam. On the other hand, the number of spawning waters was generally lower than that of Baek-jam, but hatching rate was similar to that of Baek-jam.

Therefore, in the present invention, Nd mutant silkworms were selected among the mutant strains of cordyceps as the cultivation host of Cordyceps sinensis.

The Nd mutant silkworm of the present invention was obtained from Dr. Akio Murakami, National Institute of Genetics, Japan.

In the method of artificial cultivation of Cordyceps sinensis of the present invention, the spores of Cordyceps fungi diluted to a concentration of about 10 ⁷ to 10 ⁸ spores / ml are spray-inoculated on 5th larvae of Nd mutant silkworms and subjected to 26-30 ° C. and 95% relative humidity conditions. After 12 hours or 24 hours under the protection, it is bred in mulberry according to the conventional method, when the infection is confirmed, characterized in that the fruiting body is formed by protecting at 21-25 ℃, 95% relative humidity conditions.

On the other hand, cordyceps fungi used in the method of the present invention are not particularly limited, and those skilled in the art can appropriately select and use.

[Test Example 1]

Pesylomyces japonica spores diluted at concentrations of 10 ⁷ , 5 × 10 ⁷ and 10 ⁸ spores / ml were spray-inoculated onto Nd mutant silkworms and white jade, respectively, at 5 larva stages at 28.5 ° C. and 95% relative humidity. After 12 hours and 24 hours of protection, the broth was bred according to the conventional method, and the insecticidal fungus infection rate was examined and the results are shown in FIG. 1. Infection chrysalis were protected under 25 ° C. and 95% relative humidity to form fruiting bodies, and then the rate of fruiting body formation was examined. The results are shown in FIG. 2.

From FIG. 1 and FIG. 2, when the cordyceps was grown based on Nd mutant silkworms, when the silkworms inoculated with 10 ⁸ spores / ml of Cordyceps sinensis were protected for 12 hours, the infection rate was similar to that of Baekjam. It can be seen that the fruiting body formation rate is superior to that of white jade.

On the other hand, in the production of Cordyceps sinensis (300 heads), when the Nd mutant silkworm is the cultivating host, the dry weight was 90 g, which is smaller than that of 150 g based on the white jade. This is because, as can be seen in the solution, the size of Nd (1.4 g) is smaller than that of white jade (1.7 g).

Accordingly, the present inventors hybridized Nd with Japanese 123 or Chinese 124 to improve the production of cordyceps by increasing the size of the larvae while maintaining the characteristics of nascent in producing cordyceps using Nd mutant trait. . As a result, as can be seen in Table 2, the first hybrid of Nd has a solubilization rate of 72 to 89%, and the cocoons of the remaining 11 to 28% of silkworms are thin and the tear layer is easily torn by hand. In addition, the body weight is also greatly enhanced, heavier than white jade, and the number of spawning and hatching is similar to the white jade.

Solubilization, Solubility and Hatching Ratios of Nd Mutant Silkworm Hybrids Gongsinue ¹ Dissolution Rate (%) Unavailability (%) Weight (g) Spawning water Incubation rate (%) Nd X sleep 123 97.9 73.2 2.0 386 91.7 Sleep 123 × Nd 97.8 88.7 2.1 490 93.3 Nd X sleep 124 97.9 71.6 2.0 459 96.5 Sleep 124 × Nd 97.2 77.3 2.1 531 89.8 Nd 95.3 100.0 1.2 376 91.2 White jade 99.3 0.0 1.7 434 92.7 1. Announcement: 150 heads per system

Therefore, in the present invention, one hybrid hybrid of Nd mutant silkworm was selected as the cultivation host of Cordyceps sinensis.

The method of artificially cultivating Cordyceps sinensis using one hybrid of Nd mutant silkworms is the same as that of artificial cultivation based on Nd mutant silkworms.

[Test Example 2]

Spores of Pesylomyces japonica diluted to a concentration of 10 ⁸ spores / ml were collected from one of the Japanese breeding species (Nd × Jam 123, Jam123 × Nd) and the Chinese breeding species (Nd × Jam 124) of Nd mutant silkworms. , 124 × Nd) of 5 age larvae sprayed and protected for 12 hours and 24 hours under conditions of 28.5 ℃, 95% relative humidity, and then bred in mulberry according to the conventional method, and examined the infection rate of cordyceps fungi. 3 is shown. Infection chrysalis were protected at 21 ° C. and 95% relative humidity to form fruiting bodies, and then the rate of fruiting body formation was investigated. The results are shown in FIG. 4.

3 and 4, after inoculating the first hybrid species of Nd mutant silkworms and incubating for 12 hours, it can be seen that the infection rate was 92-96% and the fruiting body formation rate was 90-96%. . In other words, it can be seen that the inoculation of Bacillus subtilis into the first hybrid of the Nd mutant silkworms shows better infection rate and fruiting body formation rate than the inoculation of Bacillus subtilis into Bacillus.

[Test Example 3]

The production amount (g / 100 head) of Cordyceps sinensis produced in Test Example 3 was measured, and the results are shown in Table 3.

Cordyceps Sinensis Production of One Hybrid Nd Mutant Silkworm (Unit: g / 100 head) Inoculation condition (time) Nd X sleep 123 Sleep 123 × Nd Nd X sleep 124 Sleep 124 × Nd Nd White jade 12 189 211 193 239 121 108 24 20 - 141 - 56 135

As shown in Table 3, the production of Cordyceps sinensis was 189 ~ 239g for Nd hybrids which showed higher value at 12 hours compared to 135g of white jade, which was higher at 24 hours. It was confirmed that Cordyceps sinensis production was increased by 40 ~ 77% compared to the method of sleeping.

On the other hand, the total labor release time required for the production of one box of cocoon was 85.3 hours, and the labor input per work stage was 18.8 hours (22%) for mulberry field management, 39.1 hours (46%) for silkworm breeding, and 16.2 hours for foot and shoulder dogs ( 19%), and other 11.2 hours (13%) (Labor time by work stages per crop, Rural Development Administration, 1996), when Cordyceps is grown using one hybrid of Nd mutant silkworms and Nd mutant silkworms. The labor costs for the foot and sowing work and the cocoon cutting are eliminated, saving 34% of the total labor for cordyceps production.

Working time by Cordyceps cultivation process (Unit: hour / box) Work content White jade sleep Nayong 1 hybrid Foot, subtraction 8.5 0.0 Dog collection 6.1 0.0 Lookout 24.0 1.2 Breeding and Others 70.7 70.7 system 109.3 71.9 (34% reduction) Some common labor requirements, such as strain inoculation, infection identification, and mushroom harvesting, were not included2. Conventional work required for sleeping sheep: 85.3 hours

As described above, artificial cultivation of Cordyceps sinensis using the first hybrid of Nd mutant silkworms and Nd mutant silkworms can reduce about one third of the labor required to grow cordyceps into general hybrids. The fruiting body formation rate is improved to increase the production of Cordyceps sinensis by an average of 54% (Fig. 5).

Claims (3)

In artificial cultivation of Cordyceps sinensis using silkworm as host insect, it is used as host insect ( ) Cultivation method characterized in that using the mutant trait (Nd). 2. The method according to claim 1, wherein one host hybrid of a mutated trait (Nd) is used as a host insect. The insecticidal fungus spores diluted at a concentration of 10 ⁷ to 10 ⁸ spores / ml are spray-inoculated to 5 larvae of host insects and subjected to spraying at 26 to 30 ° C. and 95% relative humidity for 12 hours. After protecting for a while, breeding with mulberry according to the conventional method, when the infection is confirmed, characterized in that the fruiting body is formed by protecting at 21-25 ℃, 95% relative humidity conditions.