KR101698316B1 - Method of producing sexual propagation of tubers using Ex vitro germination of Gastrodia elata seeds - Google Patents

Abstract

The present invention relates to a method for culturing seeds, comprising the step of seeding a seed using germinating Mycena spp. Germinating the root diameter from the germination of the seed of the gum using the inoculated leaves; To a method for producing seedling germination in an out-of-situ seedling for the purpose of producing a seed germ from the seed of the germ of seeds.

Description

[0001] The present invention relates to a method for producing ginseng seeds using ginseng seeds,

More particularly, the present invention relates to a method for producing germinating seeds, which comprises the steps of preparing a cotyledon litter leaf culture medium for promoting germination of the seeds of ginseng, The present invention relates to a method for producing an ovine reproduction breeder through an outgassing method of a chewing gum seed including an air-conditioning culture step of a chewing gum seed.

Chimma is a perennial higher plant belonging to the Orchidaceae and degraded in photosynthesis capacity. It has been used as a preventive and remedy for tofu disease because it is effective in paralysis, hypertension, dizziness and neurological diseases.

In 1911, the Chamaese mushroom ( Armillaria mellea) and symbiotic must first were revealed (Kusano, 1911, J Agric Coll Tokyo, 4, 1 ~ 65), since 1980, Pegasus Seeds were reported studies that have germinated by her wrinkled mushrooms (Mycena spp.) ( Xu and Guo, 1989, Acta Mycologica Sinica, 8, 221-226).

In 1995, it was possible to cultivate large quantities by the development of Chunma microbial strain No. 1 in Korea. However, due to the continuing silent propagation by Jama (hemp), the degradation phenomena of Chunma, misuse of microbial strains and lack of culture technology, And vulnerability to economic losses of farmers.

In 2009, the development of in-flight germination technique of ginseng seeds made it possible to produce a disease-free ginseng breeding gill (Korean Patent No. 10-2011-0044038) and established an artificial moisture system for stable ginseng seed harvesting (Park and Lee, 2012, Plant Biotechnol Rep., 7, 185-191). In addition, the seed germination system has been constructed using germinating microorganisms.

In addition, development of a subculture culture medium for the development of the pit diameter, analysis of useful substances, and production of in-flight microcapsules have been carried out (Korean Patent Publication No. 10-2012-0097812). In 2010, we solved the royalties problem for the Chinese lily mildew caused by the selection of Mycena sp. Strain KFRI 1212 (KACC93108P) as an in-flight cultivar.

However, it is a reality that it is very difficult for farmers to directly produce Jama by using special facilities and equipment technology.

SUMMARY OF THE INVENTION The present invention has been made in order to solve the problems of the prior art as described above, and it is an object of the present invention to provide a method and apparatus for germinating seeds of gilt- And a method for producing the same.

In order to accomplish the above object, the present invention provides a method for culturing germinated seed germs, (2) preparing a decidua rice germ culture medium for culturing an early germ culture; (3) inoculating the germ bacteria cultured in the step (1) into the decidua microflora medium prepared in the step (2); (4) preparing crushed litter medium and cut litter medium; (5) Inoculating the sprouted germs inoculated in the step (3) into the ground green leaves and cut ground leaves prepared in the step (4); And (6) co-culturing the chewing gum seeds in the pulverized litter leaf medium and the chopped litter medium inoculated in the step (5).

The present invention also provides (1) preparing a decidua rice micropowder culture for early germ culture; (2) a step of culturing the seed germs of the cheongma seed; (3) inoculating the germ bacteria cultured in the step (2) into the decidua microflora prepared in the step (1); (4) preparing a crushed litter medium and a cut litter medium; (5) Inoculating the sprouted germ inoculated in the step (3) into the ground green leaf litter and the cut green leaf medium prepared in the step (4). And (6) co-culturing the chimney seeds in the pulverized litter medium and the chopped litter medium inoculated in the step (5) above, using the outgassing method of the chimney seeds.

In addition, the present invention provides a jima produced by the above method.

The production method of the present invention enables mass production of disease-free reproductive gonads by developing a special technique, specific expensive equipment, and a method of germinating seeds of gum that have been germinated only in limited space.

Fig. 1 shows the use of a 100 x 40 x 50 mm Petri dish to spread vermiculite or sand on the bottom so that the underside can not be seen easily, water is supplied, water is supplied, and seeds of chimaese are sown in the inoculated crushed leaves and cut leaves.
FIG. 2 shows the result of the round aperture in the ground and the cut-off leaves as a result of cultivating the seeds of chewing gum at 24 ° C. for two months.
Fig. 3 shows the results of observing the germination ratio in the ground leaves and cut leaves using M. osmundicola and Mycena sp. Strain KFRI 1212 (KACC93108P).

The present invention relates to a method for culturing germinated seed germs; (2) preparing a decidua rice germ culture medium for culturing an early germ culture; (3) inoculating the germ bacteria cultured in the step (1) into the decidua microflora medium prepared in the step (2); (4) preparing crushed litter medium and cut litter medium; (5) Inoculating the sprouted germs inoculated in the step (3) into the ground green leaves and cut ground leaves prepared in the step (4); And (6) co-culturing the chewing gum seeds in the pulverized litter leaf medium and the chopped litter medium inoculated in the step (5).

The present invention also provides (1) preparing a decidua rice micropowder culture for early germ culture; (2) a step of culturing the seed germs of the cheongma seed; (3) inoculating the germ bacteria cultured in the step (2) into the decidua microflora prepared in the step (1); (4) preparing a crushed litter medium and a cut litter medium; (5) Inoculating the sprouted germ inoculated in the step (3) into the ground green leaf litter and the cut green leaf medium prepared in the step (4). And (6) co-culturing the chimney seeds in the pulverized litter medium and the chopped litter medium inoculated in the step (5) above, using the outgassing method of the chimney seeds.

In the production method of the planetary breeding herb according to the present invention, the mycelium of Mycena spp. Is cut out in an aseptic condition, and then 100 ml of a residual nutrient medium is added thereto. Lt; / RTI >

In the method for producing oily breeding jasmine of the present invention, the decidua bovine medium is prepared by adding oak-leaf litter and 3% (v / v) 20% (w / v) , The leaves cut on the corn steep liquor containing the oak-leaf litter were uniformly squeezed, and then sterilized at 121 ° C for 90 minutes.

In the method for producing the ovine breeding jasmine of the present invention, in the step of inoculating the decidua microflora culture medium, the mycelia of Mycena spp. Are aseptically cut into a sterilized deciduous microflora culture medium, , And the cells were inoculated with 5 ml of the mushroom strain, which was cultured at 27 ± 1 ° C for 3 weeks with shaking at 100 rpm, and cultured in a culture room at 27 ± 1 ° C.

In the method for producing oily breeding jima of the present invention, the crushed litter leaf medium and the cut off litter medium are prepared by adding 2% (w / w) of leaves to the cut leaves cut into 4-5 cm size of crushed and oak leaves, v) rice bran solution and sterilized at 121 캜 for 270 minutes.

The step of inoculating the crushed litter medium and the cut litter medium in the method of producing meteoric breeding herb according to the present invention comprises adding the sterilized crushed litter medium and the cut litter medium to the litter microbial culture medium at a temperature of 27 ± 1 ° C for 60 to 90 Lt; / RTI > days.

In the method for producing oily breeding jima in the present invention, the step of culturing the air conditioning may include culturing on a wet vermiculite or wet sand uniformly laid on the bottom of a Petri dish, a ground leaf litter medium, Can be sown.

In the method for producing oily breeding jima in the present invention, the step of co-culturing is a step of culturing the germinating rice seeds and the gum seeds on a petri dish on which the seedlings are seeded by immersing in water, Next, the lid of the petri dish is closed; And culturing the lid-closed petri dish at 24 +/- 1 DEG C for 4-8 weeks.

In addition, the present invention provides a jamae produced by the method of producing the female reproductive gonad using the outgassing method of the gonad seed.

The Jamae of the present invention is characterized in that it is a disease-free reproductive blastomere because it is produced using the germination method of gingko seeds.

Hereinafter, the present invention will be described in detail with reference to examples. However, these are for the purpose of illustrating the present invention in more detail, and the scope of the present invention is not limited thereto.

< Example  1> for germinating seed germination Sprouting bacteria  Production of medium

For use as a liquid culture material, Mycena spp. Cultured on a Potato Dextrose Agar (PDA, Difco; 39 g / l) medium was aseptically chopped into a 250 ml Erlenmeyer flask, 1 g / l of glucose, 0.1 g / l of glutamic acid, 2 g / l of KH2PO4, 0.5 g / l of MgSO4, 0.2 g / l of disodium dihydrate EDTA, 0.1 g / l of MnSO4, l) was added, and the cells were cultured with shaking at 27 占 1 占 폚 for about 3 weeks at 100 rpm.

The cultured mycelia were homogenized, and then 5 ml of the cells were inoculated into 15 ml of deciduous microbial broth (broth: water = 1: 4) and cultured at 27 ± 1 ° C for 3 weeks.

2% (w / v) rice bran and distilled water were added to the leaves cut into 4 ~ 5 cm size of the leaves of oak leaves and the leaves were poured into a culture container containing the inoculated sprouted bacteria, And cultured at 27 ± 1 ° C for 12 weeks.

< Example  2> Leaves Germ bacteria  Used Outdoors  germination

Using a 100x40x50 mm petri dish, the vermiculite or sand was laid on the floor only to the underside and water was kept to maintain the humidity.

The sprouted germs and the germinated seeds grown on the ground leaves and cut leaves were mixed and placed on top of them to a depth of about 2 cm.

Fig. 1 shows the seeding of chimpanzee seeds in the inoculated crushed leaves and chopped litter.

In order to maintain the water content, the leaves were soaked in water for one day and then sterilized for 90 minutes. The seeds were germinated at 24 ± 1 ° C for one month.

Fig. 2 shows the results of a circle diameter generated in the crushed litter and the cut litter medium.

< Example  3> Germination  Comparison of germination rate of ginseng seeds by germination medium

Mycelia ( M. osmundicola ) and new wrinkle mushroom ( Mycena sp . When the germinating seeds were germinated by using KFRI 1212 (KACC93108P) strain, the rate of inoculation of the leaves was faster than that of the fresh myrrh mushroom strain in M. osmundicola , 3, germination rate and M. osmundicola were 1.7 times higher.

In the litter - leaf culture medium, only the Liliaceae spp. Germinated was germinated.

Although the present invention has been described in connection with the specific embodiments of the present invention, it is to be understood that the present invention is not limited thereto. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

In addition, the materials of each of the constituent elements described herein can be easily selected and substituted for various materials known to those of ordinary skill in the art. In addition, a person skilled in the art will be able to add components to omit some of the components described herein without degradation of performance or to improve performance.

In addition, a typical technician may change the order of the method steps described herein depending on the process environment or equipment. Accordingly, the scope of the present invention should be determined not by the embodiments described, but by the claims and their equivalents.

According to the method for producing oily breeding jima using the out-germination method of the present invention, there is an advantage that seed germination can be promoted from outside of the ovary to root diameter.

Therefore, it is possible to generalize the production method of meteoric breeding jasmine by overcoming the degradation phenomenon of Chunma by the existing asex breeding cultivation method and through the specific culture facility and technology.

Claims (9)

(1) culturing Mycena osmundicola as a germinating seed of gilt seed;
(2) preparing a decidua rice germ culture medium for culturing an early germ culture;
(3) inoculating the germ bacteria cultured in the step (1) into the decidua microflora medium prepared in the step (2);
(4) preparing crushed litter medium and cut litter medium;
(5) Inoculating the sprouted germs inoculated in the step (3) into the ground litter medium and the cut litter medium prepared in the step (4), wherein the step of inoculating the ground litter medium and the cut litter medium comprises: Adding crushed litter medium and cut litter medium to deciduous rice gonad medium and culturing at 27 +/- 1 DEG C for 60 to 90 days; And
(6) A method for producing oily breeding jima using the outgassing method of chick horse seeds, comprising co-culturing the chimney seeds in the pulverized litter leaf medium and the cut litter medium inoculated in the step (5). (1) preparing a decidua rice germ culture medium for culturing an early germ culture;
(2) a step of culturing Mycena osmundicola as a germinating seed germ,
(3) inoculating the germ bacteria cultured in the step (2) into the decidua microflora prepared in the step (1);
(4) preparing a crushed litter medium and a cut litter medium;
(5) The step of inoculating the sprouted germ inoculated in the step (3) into the pulverized litter leaf medium and the cut litter medium prepared in the step (4), wherein the step of inoculating the pulverized litter leaf medium and the cut litter medium , Adding sterilized ground lit leaf medium and cut lit leaf medium to deciduous rice gonad medium and culturing at 27 占 폚 for 60 to 90 days; And
(6) A method for producing an ovine breeding jasmine by using an outgassing method of chick horse seeds, comprising co-culturing the chick horse seeds in the pulverized litter leaf medium and the cut litter medium inoculated in the step (5). 3. The method according to claim 1 or 2,
The germinating microorganism is obtained by cutting off the mycelia of Mycena osmundicola in an aseptic condition, adding 100 ml of a minnow medium, and culturing the mixture at 27 ± 1 ° C for 3 weeks with shaking at 100 rpm. Production method of meteoric breeding jama using germination method. 3. The method according to claim 1 or 2,
The decidua rice gravel medium was prepared by adding oak-leaf litter to a culture vessel and adding 20% (w / v) rice bran solution containing 3% (v / v) of the volume of the culture vessel to the culture vessel, A method for producing oily breeding jima using the outgassing method of a chewing gum seed, characterized in that the leaves are homogenously levitated and then sterilized at 121 캜 for 90 minutes. 3. The method according to claim 1 or 2,
In the step of inoculating the decidua microflora medium, the mycelia of the Mycena osmundicola strain were aseptically cut into the sterilized deciduous microflora medium, 100 ml of the fineness medium was added, and the mixture was cultured at 100 占 폚 for 3 weeks at 27 占 폚 A method for producing oily breeding jima using the germination method of chick horse seeds, which comprises culturing in a culture room of 27 ± 1 ° C inoculated with 5 ml of a shake cultivated wilting mushroom strain. 3. The method according to claim 1 or 2,
2% (w / v) corn steep liquor was added to the cut leaves cut into 4 ~ 5 cm size of crushed leaves and oak leaves which were crushed by oven deciduous leaves and crushed green leaves, Wherein said germinating seeds are sterilized for a period of time equal to or longer than a predetermined time. delete 3. The method according to claim 1 or 2,
The step of culturing the air conditioning includes: seeding germinating bacteria and germinating seeds cultured in a ground leaf litter medium and a ground litter medium on wet vermiculite or wet sand uniformly spread on the bottom of a Petri dish;
Dipping the seed germs and the germinated seeds in a petri dish on which the seeds have been seeded, covering the cut leaves sterilized at 121 캜 for 90 minutes, and then closing the lid of the petri dish; And
Culturing the petri dish with the lid closed at 24 +/- 1 DEG C for 4 to 8 weeks, wherein the petri dish is cultured at 24 +/- 1 DEG C for 4 to 8 weeks. delete

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