KR20200066577A - A Method for mars production of gametophyte of brown seaweed - Google Patents

A Method for mars production of gametophyte of brown seaweed Download PDF

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KR20200066577A
KR20200066577A KR1020190157783A KR20190157783A KR20200066577A KR 20200066577 A KR20200066577 A KR 20200066577A KR 1020190157783 A KR1020190157783 A KR 1020190157783A KR 20190157783 A KR20190157783 A KR 20190157783A KR 20200066577 A KR20200066577 A KR 20200066577A
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seaweed
alginate
spores
culture
concentration
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KR102364453B1 (en
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신현웅
정상목
윤정현
박태희
한승희
이창연
전원빈
박지현
황동수
정성준
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순천향대학교 산학협력단
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G33/00—Cultivation of seaweed or algae
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • A01C1/06—Coating or dressing seed
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/88—

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Ecology (AREA)
  • Forests & Forestry (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Soil Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Botany (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Cultivation Of Seaweed (AREA)

Abstract

The present invention relates to a technique, which cultivates extracted seaweed spores to preserve and proliferate the same to a gametophyte through temperature and illumination control, grows adults by converting the proliferated gametophyte to sporophyte, and forms a seaweed colony after coating the proliferated gametophyte with alginate, applying the same to an offshore structure, and cultivating the same.

Description

미역 배우체의 대량 생산 방법{A Method for mars production of gametophyte of brown seaweed}A method for mars production of gametophyte of brown seaweed}

본 발명은 미역 배우체를 실험실 내에서 온도 및 조도 조절을 통해 인위적인 휴면 상태를 만들어 암,수 배우체의 수정을 방지하고 각각을 증식시켜 미역 배우체를 대량생산하는 방법에 관한 것이다. The present invention relates to a method for mass production of seaweed spores by preventing the fertilization of male and female spouses by multiplying each of them by creating an artificial dormant state by controlling temperature and illuminance in the laboratory.

또한 상기의 미역 배우체를 피막화하여 미역 배우체의 착생을 개선시켜 해양 구조물에 도포 및 착생시킨 후 해역 온도에서 배우체가 다시 활성 상태로 전환하여 수정과 아포체 생성 및 미역 군락을 조성하는 기술에 관한 것이다.In addition, it relates to a technique for improving the engraftment of seaweed spores by encapsulating the above seaweed spores, applying them to marine structures, and then converting the spores back to an active state at sea temperature to create fertilization and spore formation and seaweed colonies. .

해중림, 즉 바다숲은 해양 수중으로 산소를 공급하고 해양 환경을 정화하며, 각종 어패류의 산란장과 성육장을 제공하여 해양 생태계의 일차 생산자로서 그 기능이 매우 중요하다. The marine forest, that is, the sea forest, provides oxygen to the underwater water, purifies the marine environment, and provides a spawning and breeding ground for various fish and shellfish, so its function as a primary producer of the marine ecosystem is very important.

그러나 최근 들어, 국내 연안 어장에서 무성했던 미역, 다시마, 감태, 모자반, 잘피 등의 다년생 해조류 군락이 고수온, 저염분, 해양오염, 조식성 동물의 과다 섭생 등으로 생태계가 파괴되어 갯녹음 현상(즉, 백화현상)이 발생되고 있다. 이러한 백화현상은 현재 동해안 일대에 급속하게 확산되어 연안 양식어장이 황폐화되고 있으며 미역, 다시마 등의 해중림도 급격히 파괴되어 전복, 성게 등의 먹이 사슬 구조 변화와 각종 어류의 산란 및 서식환경을 포함한 해양생태계에 상당한 영향을 미치고 있는 실정이다. 백화현상은 여러 언론기관을 통해서 보도된 바와 같이, 동해안 전역에서 발생하고 있고, 연안 10m 내외의 수심까지 확산되고 있으며, 지금까지 총 어장의 넓은 면적에서 발생하여 어민들에게 큰 피해를 입히고 있는 있다. 그러나, 한번 백화현상이 발생된 해역에서는 수산자원의 고갈은 물론 부영양화도 촉진되어, 이를 인위적으로 회복시키는 노력이 없이는 자연적인 치유가 불가능한 것으로 알려져 있다. In recent years, however, the perennial seaweed communities such as seaweed, kelp, persimmon, capsicum, and jalpii, which flourished at domestic coastal fishing grounds, have been destroyed by ecosystems due to high temperature, low salinity, marine pollution, and excessive diet of breakfast animals (ie Whitening phenomenon). This whitening phenomenon is rapidly spreading around the East Coast, causing coastal fishery to be desolated, and marine forests, such as seaweed and kelp, are also rapidly destroyed, changing the structure of the food chain such as abalone and sea urchins, and the marine ecosystem, including various fish spawning and habitat environments. It is a situation that has a significant impact on the. As reported by various media organizations, the whitening phenomenon is occurring all over the East Coast, spreading to the depths of about 10m off the coast, and so far, has occurred in a large area of total fishing grounds, causing great damage to fishermen. However, it is known that natural healing is not possible without efforts to artificially restore the depletion of fishery resources as well as the promotion of eutrophication as well as the depletion of fisheries resources in the sea area where whitening occurred once.

인위적으로 포자를 이식하여 바다숲을 조성할 수 있으나, 포자를 이식하기 위해 미역 암수 배우체 분리시 분리과정이 번거로우며, 불완전한 분리로 인해 수정되는 문제가 있다.Although artificial spores can be transplanted to form a sea forest, the separation process is cumbersome when separating seaweed male and female spores for transplanting spores, and there is a problem that is corrected due to incomplete separation.

대한민국 공개특허 제10-2010-0059042호Republic of Korea Patent Publication No. 10-2010-0059042

본 발명의 목적은 미역(Undaria pinnatifida) 배우체를 25 내지 27℃의 온도에서 1000 내지 2200 lx의 조도 조건으로 배양하여 휴면상태로 유지시키는 단계를 포함하는, 미역 배우체의 대량 생산방법을 제공하는 것이다. An object of the present invention is to provide a method for mass production of seaweed spores, comprising the step of culturing seaweed (Undaria pinnatifida ) spores at a temperature of 25 to 27°C under an illumination condition of 1000 to 2200 lx to maintain a dormant state.

본 발명의 다른 목적은 상기 생산방법으로 생산한 휴면상태의 미역 배우체를 105 내지 107 cells/㎖의 농도로 포함하는 배양액을 제조하는 단계, 상기 배양액 100 중량부에 대하여 알지네이트를 0.2 내지 2 중량부로 혼합하여 혼합물을 제조하는 단계 및 상기 혼합물에 다가 금속염을 첨가하여 경화시켜 피막화된 미역 배우체를 제조하는 단계를 포함하는, 미역 배우체의 착생 개선 방법을 제공하는 것이다. Another object of the present invention is a step of preparing a culture medium containing a concentration of 10 5 to 10 7 cells/ml of a seaweed spouse in a dormant state produced by the production method, 0.2 to 2 weight of alginate per 100 parts by weight of the culture medium It is to provide a method for improving the engraftment of seaweed spores, comprising the steps of mixing the parts to prepare a mixture, and adding a polyvalent metal salt to the mixture to cure to prepare a coated seaweed spore.

본 발명은 기존의 미역 암,수 배우체 분리 시 문제점인 분리과정의 번거로움과 불완전 분리로 인한 수정 및 아포체 생성을 방지함으로써 더욱 용이한 배우체 분리배양을 위하여 안출된 것이다.The present invention has been devised for easier separation and culture of spores by preventing fertilization and sporulation due to incomplete separation and hassle of separation process, which is a problem when separating seaweed cancer and male spores.

본 발명의 발명자들은 미역 배우체를 실험실 내에서 온도 및 조도 조절을 통해 인위적인 휴면 상태를 만들어 암,수 배우체의 수정을 방지하고 각각을 증식시키는 미역 배우체의 배양 조건을 확인하였다. 또한 상기 휴면 상태로 증식된 미역 배우체를 알지네이트(Alginate)로 피막화하여 미역 배우체의 착생 성장 개체수를 개선시키는 피막화 조건을 확인하였다. The inventors of the present invention confirmed the cultivation conditions of seaweed spouses to prevent the fertilization of male and female spouses and proliferate each of them by creating an artificial dormant state by adjusting temperature and illuminance in the laboratory. In addition, it was confirmed that the encapsulation conditions for improving the number of seaweed spouses growing by growing the seaweed spores grown in the dormant state with Alginate.

상기 피막화된 미역 배우체를 해양 구조물에 도포 및 착생시킨 후 해역 온도에서 배우체가 다시 활성 상태로 전환하여 수정과 아포체 생성 및 미역 군락을 조성할 수 있다.After the coated seaweed spores are applied to the marine structure and grown, the spores can be converted back to an active state at sea temperature to create fertilization, spore formation, and seaweed colonies.

상기와 같은 목적을 달성하기 위해 본 발명은 하나의 양태로 미역(Undaria pinnatifida) 배우체를 25 내지 27℃의 온도에서 1000 내지 2200 lx의 조도 조건으로 배양하여 휴면상태로 유지시키는 단계를 포함하는, 미역 배우체의 대량생산 방법을 제공한다. In order to achieve the above object, the present invention includes the step of culturing a seaweed ( Undaria pinnatifida ) spouse in an embodiment at a temperature of 25 to 27°C under an illumination condition of 1000 to 2200 lx to maintain a dormant state. It provides a method for mass production of spouses.

본 발명의 미역 배우체의 대량생산 방법은 미역(Undaria pinnatifida) 배우체를 25 내지 27℃의 온도에서 1000 내지 2200 lx의 조도 조건으로 배양하여 휴면상태로 유지시키는 단계를 포함한다.The method for mass production of seaweed spores of the present invention includes the step of culturing seaweed ( Undaria pinnatifida ) spores at a temperature of 25 to 27°C under an illumination condition of 1000 to 2200 lx to maintain a dormant state.

본 발명의 실시예에서 미역 배우체를 1000 lx의 조도에서 명암 주기 12:12로 20 내지 30℃의 온도조건에서 배양한 후, 아포체 형성률과 배우체 성장율을 분석하였다.In the embodiment of the present invention, after the seaweed spores were cultured at a temperature of 20 to 30°C with a contrast period of 12:12 at an illuminance of 1000 lx, the spore formation rate and spore growth rate were analyzed.

분석결과, 온도가 가장 낮을수록 배우체 성장률이 우수함을 확인하였다. 구체적으로 20℃에서 배우체 성장율이 90.47%로 가장 높았으며, 30℃에서 배우체 성장율이 11.03%로 현저하게 낮아짐을 확인하였다. 또한, 온도가 낮을수록 배우체의 수정을 통한 아포체 형성율이 가장 우수함을 확인하였고, 26℃ 이상에서는 배우체가 수정되어 아포체를 형성하는 단계가 진행되지 않음을 확인하였다. 즉, 26℃ 에서 배양시 미역 배우체의 수정을 방지하여 아포체 형성이 낮으며, 동시에 배우체 성장율이 높아 미역 배우체를 대량생산하는데 적합함을 확인하였다. As a result of the analysis, it was confirmed that the lower the temperature, the better the growth rate of the sperm. Specifically, it was confirmed that the sperm growth rate was the highest at 20°C at 90.47%, and the sperm growth rate at 30°C was remarkably lowered to 11.03%. In addition, it was confirmed that the lower the temperature, the better the rate of spore formation through fertilization of the spore, and above 26°C, it was confirmed that the step of forming the spore by modifying the spore does not proceed. That is, when cultured at 26°C, it was confirmed that it is suitable for mass production of seaweed spores by preventing fertilization of the seaweed spores, and thus having low spore formation and high growth rate of spores.

따라서 본 발명의 미역 배우체 대량 생산방법은 미역(Undaria pinnatifida) 배우체를 25 내지 27℃의 온도에서 배양되는 것이며, 구체적으로 26℃에서 배양되는 것 일수 있다. Therefore, the method for mass production of seaweed spores of the present invention is that the seaweed ( Undaria pinnatifida ) spores are cultured at a temperature of 25 to 27° C., and may be specifically cultured at 26° C.

상기 생산방법에 의해 생산된 미역 미역 배우체는 25 내지 27℃의 온도에서 배양하여 암, 수 배우체가 수정이 진행되지 않아 아포체를 형성되지 않으며, 미역 배우체의 휴면상태를 유지한다.Seaweed seaweed spores produced by the above-described production method are cultured at a temperature of 25 to 27° C., so that male and female spouses do not undergo fertilization, so that spores are not formed, and the seaweed spores maintain a dormant state.

본 발명에서 아포체는 암 배우체와 수 배우체의 생식의 결과로 포자를 형성하는 식물체를 의미한다. In the present invention, the apoce refers to a plant that forms spores as a result of reproduction of the cancerous and male partners.

미역 배우체의 배양온도가 25℃ 미만에서는 암 배우체와 수 배우체가 서로 수정하여 아포체를 형성하는 문제가 있으며, 27℃ 초과에서는 배우체의 성장율이 낮아 배양의 효율성이 떨어지는 문제가 있다 When the culture temperature of the seaweed sperm is less than 25°C, there is a problem that the female sperm and the male sperm fertilize each other to form a spore, and when it exceeds 27°C, the growth rate of the sperm is low, so the efficiency of culture decreases.

본 발명의 다른 실시예에서 미역 배우체를 조도조건을 1200 내지 3200 lx로 달리하여 26℃, 명암 주기 12:12로 배양하고 배우체 성장율을 비교한 결과, 1200에서 2000 lx로 조도가 증가함에 따라 80.45% 에서 84.64%로 배우체 성장율이 증가하였으며, 2000 lx를 초과하는 경우 배우체 성장율이 감소하는 것을 확인하였다. In another embodiment of the present invention, the seaweed spores were cultured at 26° C., a contrast cycle of 12:12 with different illumination conditions of 1200 to 3200 lx, and the growth rate of the spores was compared to 80.45% as the illuminance increased from 1200 to 2000 lx. At 84.64%, the growth rate of the sperm increased, and when it exceeded 2000 lx, the sperm growth rate decreased.

본 발명의 미역 배우체 대량 생산방법은 미역(Undaria pinnatifida) 배우체를 1000 내지 2200 lx 조도 조건에서 배양하며, 구체적으로 2000 lx의 조도 조건으로 배양하는 것일 수 있다. The method for mass production of seaweed spores of the present invention is to culture seaweed ( Undaria pinnatifida ) spores under conditions of 1000 to 2200 lx roughness, and may be specifically cultivated under roughness conditions of 2000 lx.

상기 생산방법에 의해 생산된 미역 미역 배우체는 1000 내지 2200 lx 조도 조건에서 배양하여 배우체 성장율이 우수하여, 미역 배우체를 효율적으로 대량생산이 가능하다. Seaweed seaweed spawn produced by the above production method is cultivated under conditions of 1000 to 2200 lx roughness, so that the growth rate of spores is excellent, and it is possible to efficiently mass-produce seaweed spores.

본 발명의 실시예에서 미역 배우체를 광주기는 12:12(명기: 암기)로 배양하였다. 따라서 상기 미역 배우체는 광주기 조건 12:12(명기: 암기)로 배양되는 것일 수 있다. In the embodiment of the present invention, the seaweed spouse was cultured at 12:12 (memory: memorization). Therefore, the seaweed spouse may be cultured under Gwangju period condition 12:12 (memory period: memorization).

본 발명의 다른 양태로 상기 생산방법으로 생산한 휴면상태의 미역 배우체를 105 내지 107 cells/㎖의 농도로 포함하는 배양액을 제조하는 단계, 상기 배양액 100 중량부에 대하여 알지네이트를 0.6 내지 1.8 중량부로 혼합하여 혼합물을 제조하는 단계 및 상기 혼합물에 다가 금속염을 첨가하여 경화시켜 피막화된 미역 배우체를 제조하는 단계를 포함하는, 미역 배우체의 착생 개선 방법을 제공한다. In another aspect of the present invention, a step of preparing a culture medium comprising a dormant seaweed spore produced in the production method at a concentration of 10 5 to 10 7 cells/ml, wherein alginate is 0.6 to 1.8 weight per 100 parts by weight of the culture medium. It provides a method for improving the engraftment of seaweed spores, comprising the steps of mixing the parts to prepare a mixture, and adding a polyvalent metal salt to the mixture to cure to prepare a coated seaweed spore.

본 발명의 미역 배우체의 착생 개선 방법은 상기 생산방법으로 생산한 휴면상태의 미역 배우체를 105 내지 107 cells/㎖의 농도로 포함하는 배양액을 제조하는 단계, 상기 배양액 100 중량부에 대하여 알지네이트를 0.6 내지 1.8 중량부로 혼합하여 혼합물을 제조하는 단계 및 상기 혼합물에 다가 금속염을 첨가하여 경화시켜 피막화된 미역 배우체를 제조하는 단계를 포함한다.The method for improving the growth of seaweed spores of the present invention comprises the steps of preparing a culture medium containing a dormant seaweed spore produced in the production method at a concentration of 105 to 107 cells/ml, and alginate 0.6 to 100 parts by weight of the culture medium. It comprises mixing 1.8 parts by weight to prepare a mixture, and adding a polyvalent metal salt to the mixture to cure to prepare a coated seaweed spouse.

본 발명에서 상기 피막화된 미역 배우체를 착생이 개선되어 밧줄 또는 그물 발에 부착시켜 미역 양식에 이용할 수 있다. In the present invention, the filmed seaweed spouse is improved in adhesion and can be used in seaweed culture by attaching it to a rope or a net foot.

본 발명에서 상기 미역 배우체는 25 내지 27℃의 온도에서 1000 내지 2200 lx의 조도 조건으로 배양되어 암, 수 배우체가 수정이 진행되지 않고 휴면상태를 유지하고 있는 것을 특징으로 한다. In the present invention, the seaweed spouse is cultured at a temperature of 25 to 27°C under an illuminance condition of 1000 to 2200 lx, and the male and female spouses maintain a dormant state without fertilization.

상기 미역 배우체는 피막화를 통해 해양 구조물, 밧줄, 그물발 등에 도포시 착생이 개선되며. 해역온도(25℃ 미만)에서 활성 상태로 전환되어 수정과 아포체 형성을 할 수 있다. 따라서 미역 군락 형성에 유용하며, 미역 양식에 활용할 수 있다.The seaweed spouse is improved in sea life when applied to marine structures, ropes, nets, etc. through encapsulation. It can be converted into an active state at a seawater temperature (below 25°C) to form crystals and spores. Therefore, it is useful for forming seaweed communities and can be used for seaweed farming.

본 발명에서 상기 다가 금속염은 염화칼슘, 염화마그네슘, 염화철(2가), 염화철(3가), 황산칼슘, 황산마그네슘, 황산철(2가), 황산철(3가)로 이루어진 군에서 선택되는 1종 이상일 수 있다. In the present invention, the polyvalent metal salt is selected from the group consisting of calcium chloride, magnesium chloride, iron chloride (divalent), iron chloride (trivalent), calcium sulfate, magnesium sulfate, iron sulfate (divalent), iron sulfate (trivalent) 1 It may be more than a species.

본 발명의 실시예에서 경화제로 염화칼슘을 3 g/100㎖를 첨가하였다.In the examples of the present invention, 3 g/100 ml of calcium chloride was added as a curing agent.

따라서 상기 다가 금속염은 구체적으로 염화칼슘일 수 있다. Therefore, the polyvalent metal salt may be specifically calcium chloride.

또한, 상기 피막화된 미역 배우체를 제조하는 단계는 상기 배양액 100 중량부에 대하여 다가 금속염을 3 중량부를 첨가하는 것일 수 있다.In addition, the step of preparing the coated seaweed spouse may be to add 3 parts by weight of a polyvalent metal salt with respect to 100 parts by weight of the culture medium.

본 발명의 실시예에서 미역 배우체를 1.17×105 내지 1.17×107 cells/㎖의 농도로 포함하는 배양액에 알지네이트 수용액 0.2 내지 2g/100㎖ 첨가하고 3g/100㎖의 염화칼슘 경화제를 첨가하여 피막화된 미역 배우체를 제조하고, 1개월 후의 미역 배우체의 착생 성장 개체 수를 분석하였다.In an embodiment of the present invention, 0.2-2 g/100 ml of alginate aqueous solution is added to a culture solution containing seaweed spores at a concentration of 1.17 x 10 5 to 1.17 x 10 7 cells/ml, and 3 g/100 ml of calcium chloride curing agent is added to form a film. Prepared seaweed spouses were prepared, and the number of growth growth individuals of seaweed spouses after 1 month was analyzed.

분석결과, 미역 배우체를 1.17 × 105 내지 1.17 × 107 cells/㎖의 농도로 포함하는 배양액을 이용한 경우 모두 알지네이트의 농도가 0.8g/100㎖에서 착생 성장수가 1.62 ± 0.32 내지 2.47 ± 0.44 cells/㎠으로 가장 우수함을 확인하였다. 이중 1.17 × 106 cells 농도의 미역 배우체를 이용한 경우 착생 성장수가 2.47 ± 0.44 cells/㎠로 가장 높게 나타났다. As a result of the analysis, in the case of using a culture medium containing a seaweed spouse at a concentration of 1.17 × 10 5 to 1.17 × 10 7 cells/ml, the growth number of alginate was 1.62 ± 0.32 to 2.47 ± 0.44 cells/ at an alginate concentration of 0.8 g/100 ml. It was confirmed that the most excellent in cm 2. Among them, when seaweed spouses with a concentration of 1.17 × 10 6 cells were used, the growth number of seedlings was highest at 2.47 ± 0.44 cells/㎠.

반면, 알지네이트 수용액의 농도 0.4g/100㎖ 이하에서는 착생 성장수가 0.11 ± 0.04 cells/㎠로 측정되어 낮아 알지네이트 수용액을 0.4g/100㎖ 이하로 이용할 경우, 착생 개선 효과가 낮은 것을 확인하였다. 또한, 알지네이트의 농도가 1/100㎖ 에서 2/100㎖로 증가할수록 착생 성장수가 감소하는 경향을 나타냈으며, 동일한 미역 배우체 배양액 농도에서 비교하였을 때, 알지네이트 수용액의 농도가 2/100㎖일 때 효과가 가장 우수한 0.8g/100㎖의 비교하여 착생 성장수가 절반 수준(1.19 내지 0.55 cells/㎠)으로 감소하는 것을 확인하였다. On the other hand, when the concentration of the alginate aqueous solution was 0.4 g/100 ml or less, the number of growth of growth was measured to be 0.11 ± 0.04 cells/cm 2, and it was confirmed that when the alginate aqueous solution was used at 0.4 g/100 ml or less, the effect of improving the growth was low. In addition, as the concentration of alginate increased from 1/100 ml to 2/100 ml, the number of growth growths tended to decrease, and when compared at the same concentration of seaweed sperm culture, the effect of alginate aqueous solution concentration was 2/100 ml Compared with the most excellent 0.8g / 100ml it was confirmed that the number of growth growth is reduced to half the level (1.19 to 0.55 cells / ㎠).

따라서 본 발명의 배양액을 제조하는 단계는 미역 배우체를 1.17 × 105 내지 1.17 × 107 cells/㎖의 농도로 포함하도록 제조하는 것일 수 있으며, 구체적으로 1.17 × 106 cells/㎖의 농도로 포함하도록 제조하는 것일 수 있다. Therefore, the step of preparing the culture medium of the present invention may be to include the seaweed spouse at a concentration of 1.17 × 10 5 to 1.17 × 10 7 cells/ml, specifically to include the concentration of 1.17 × 10 6 cells/ml. It may be manufactured.

또한 본 발명에서 혼합물을 제조하는 단계는 상기 배양액 100 중량부에 대하여 알지네이트를 0.6 내지 1.8 중량부로 혼합하여 혼합물을 제조하는 것일 수 있으며, 구체적으로 100 중량부를 기준으로 0.7 내지 0.9 중량부로 혼합하는 것일 수 있고, 더욱 구체적으로 0.8 중량부로 혼합하는 것일 수 있다. In addition, the step of preparing a mixture in the present invention may be to prepare a mixture by mixing alginate in an amount of 0.6 to 1.8 parts by weight based on 100 parts by weight of the culture medium, and specifically, mixing in an amount of 0.7 to 0.9 parts by weight based on 100 parts by weight. And more specifically, may be mixed in 0.8 parts by weight.

본 발명은 미역 배우체의 대량 생산 방법에 관한 것이다. 본 발명의 방법은 미역 육종에 유용하게 사용될 수 있는 배우체 대량 증식을 통해, 미역 군락을 조성할 수 있어, 바다숲 조성에 유용하다. 또한, 미역이 대량 생산을 통해 고부가 가치를 창출하는 화장품, 의약품 및 건강식품 원료 공급에 활용될 수 있다.The present invention relates to a method for mass production of seaweed spouses. The method of the present invention can be used for seaweed breeding, it is possible to create a seaweed colony through mass growth of spouses, which is useful for sea forest formation. In addition, seaweed can be used to supply raw materials for cosmetics, medicines, and health foods that create high added value through mass production.

이하, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본 발명의 실시예에 대하여 첨부한 도면을 참고로 하여 상세히 설명한다. 그러나 본 발명은 여러 가지 상이한 형태로 구현될 수 있으며 여기에서 설명하는 실시예에 한정되지 않는다. Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art to which the present invention pertains can easily practice. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein.

<배양 온도 조건에 따른 휴면보존 및 증식 분석><Domestic preservation and growth analysis according to culture temperature conditions>

미역(Undaria pinnatifida) 배우체의 휴면 보존 또는 증식의 최적 배양 온도를 확인하고자, 실시예1-1 내지 1-6에서 미역 배우체의 배양 온도를 달리하여 미역 배우체의 성장율과 아포생성율을 분석하였다. In order to determine the optimal culture temperature of dormant preservation or proliferation of seaweed ( Undaria pinnatifida ) spores, growth rates and spore production rates of seaweed spores were analyzed by varying the culture temperature of seaweed spores in Examples 1-1 to 1-6.

실시예 1-1Example 1-1

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 20℃, 조도 1000 ㏓, 광주기를 명기: 암기(Light:Dark, L:D)= 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 20° C., an illuminance of 1000 ㏓, and a photoperiod: Light: Dark (L:D) = 12:12. .

실시예 1-2Example 1-2

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 22℃, 조도 1000 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 22° C., an illuminance of 1000 ㏓, and L:D = 12:12 to observe the growth rate of sperm after 7 days of culture.

실시예 1-3Example 1-3

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 24℃, 조도 1000 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 24° C., an illuminance of 1000 ㏓, and L:D = 12:12 to observe the sperm growth rate after 7 days of culture.

실시예 1-4Example 1-4

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 1000 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 26° C., an illuminance of 1000 ㏓, and L:D = 12:12 to observe the growth rate of the spores after 7 days of culture.

실시예 1-5Example 1-5

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 28℃, 조도 1000 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 28° C., an illuminance of 1000 ㏓, and L:D = 12:12 to observe the growth rate of the spores after 7 days of culture.

실시예 1-6Example 1-6

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 30℃, 조도 1000 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 30° C., an illuminance of 1000 ㏓, and L:D = 12:12, and the sperm growth rate was observed after 7 days of culture.

표 1은 미역 배우체 배양온도에 따른 성장률 비교한 결과이다. Table 1 is a comparison of the growth rate according to the culture temperature of seaweed sperm.

구분division 실시예
1-1Example
1-1 실시예
1-2Example
1-2 실시예
1-3Example
1-3 실시예
1-4Example
1-4 실시예
1-5Example
1-5 실시예
1-6Example
1-6 온도(℃)Temperature (℃) 2020 2222 2424 2626 2828 3030 배우체 성장률(%)Spousal growth rate (%) 90.4790.47 84.6184.61 81.2081.20 79.9879.98 45.2545.25 11.0311.03 아포체 형성률Sporulation rate ++++++++ ++++++ ++++ ++ ++ ++

(++++:100-75%, +++:74-50%, ++:49-1%, +:0%) (++++:100-75%, +++:74-50%, ++:49-1%, +:0%)

상기 표 1에서 온도가 낮을수록 배우체 성장률이 높아지는 경향을 보였으며, 26℃이상의 배양 조건에서는 배우체가 수정되어 아포체를 형성하는 단계가 진행되지 않았다.In Table 1, the lower the temperature, the higher the growth rate of the embryonic body, and in the culture condition of 26°C or higher, the step of forming the spores by fertilization of the embryonic body did not proceed.

즉, 26℃ 에서 배양시 미역 배우체의 수정을 방지하여 아포체 형성이 낮으면서, 동시에 배우체 성장율이 높아 미역 배우체를 대량생산하는데 적합함을 확인하였다. That is, it was confirmed that it is suitable for mass production of seaweed spores by preventing the fertilization of seaweed spores when incubating at 26° C., thereby preventing spore formation, while simultaneously increasing the growth rate of spores.

<조도 조건에 따른 휴면보존 및 증식 분석><Domestic preservation and proliferation analysis according to the illumination conditions>

다음으로 조도 조건에 따른 미역 배우체 보존 및 증식을 확인하고자, 실시예 1-7 내지 1-12와 같이 조도를 달리하고 미역 배우체의 성장률을 확인하였다. Next, in order to confirm the preservation and proliferation of seaweed spores according to the conditions of illumination, different illuminances were performed as in Examples 1-7 to 1-12 to confirm the growth rate of seaweed spouses.

실시예 1-7Example 1-7

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 1200 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 26° C., an illuminance of 1200 ㏓, and L:D = 12:12, and the sperm growth rate was observed after 7 days of culture.

실시예 1-8Example 1-8

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 1600 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 26° C., an illuminance of 1600 ㏓, and L:D = 12:12 to observe the growth rate of the spores after 7 days of culture.

실시예 1-9Example 1-9

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 2000 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set at a temperature of 26° C., an illuminance of 2000 ㏓, and L:D = 12:12, and the growth rate of sperm was observed after 7 days of culture.

실시예 1-10Example 1-10

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 2400 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set at a temperature of 26° C., an illuminance of 2400 ㏓, and L:D = 12:12, and the growth rate of sperm was observed after 7 days of culture.

실시예 1-11Example 1-11

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 2800 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 26° C., an illuminance of 2800 ㏓, and L:D = 12:12 to observe the growth rate of sperm after 7 days of culture.

실시예 1-12Example 1-12

미역 배우체를 멸균해수 50㎖에 PES I 배지가 첨가된 배양액이 담긴 배양 디쉬에 넣었다. 배양 챔버 안에서 이를 배양하였으며, 배양 챔버의 조건은 온도 26℃, 조도 3200 ㏓, L:D = 12:12로 설정하여 배양 7일 후 배우체 성장률을 관찰하였다.The seaweed sperm was placed in a culture dish containing a culture medium in which PES I medium was added to 50 ml of sterile seawater. This was cultured in a culture chamber, and the conditions of the culture chamber were set to a temperature of 26° C., an illuminance of 3200 ㏓, and L:D = 12:12 to observe the growth rate of the spores after 7 days of culture.

표 2는 조도에 따른 미역 배우체의 성장률 분석한 결과이다. Table 2 shows the results of analyzing the growth rate of seaweed spouses according to the roughness.

구분division 실시예
1-7Example
1-7 실시예
1-8Example
1-8 실시예
1-9Example
1-9 실시예
1-10Example
1-10 실시예
1-11Example
1-11 실시예
1-12Example
1-12 조도(㏓)Roughness 12001200 16001600 20002000 24002400 28002800 32003200 배우체 성장률(%)Spousal growth rate (%) 80.4580.45 82.7882.78 84.6484.64 77.3577.35 62.7662.76 48.1848.18

상기 표 2에서 조도 2000 ㏓의 조건에서 가장 높은 배우체의 성장이 관찰되었고 2000 ㏓를 초과하는 조도 조건에서는 성장률이 꾸준히 감소하는 경향을 보였다.In Table 2, the growth of the highest spouse was observed under the condition of 2000 조 and the growth rate tended to decrease steadily under the condition of 2000 ㏓ or more.

<알지네이트 또는 미역 배우체의 농도에 따른 미역 배우체의 착생 성장 개체 비교><Comparison of growth growth of seaweed spouses according to the concentration of alginate or seaweed spouses>

휴면 배우체를 피막화하기 위한 알지네이트(Alginate)의 농도 및 해양 구조물에 도포하여 해역에서 착생 및 성장하기 위한 최적의 미역 배우체 농도를 확인하고자, 실시예 2-1 내지 2-18과 같이 알지네이트(Alginate) 염의 농도 또는 미역 배우체의 농도를 달리하고 1개월 후 미역 배우체의 착생 성장 개체수를 비교하였다.In order to determine the concentration of alginate (Alginate) for coating the dormant sperm and the optimal seaweed sperm concentration for growth and growth in the sea area by coating the marine structure, as in Examples 2-1 to 2-18, Alginate Different salt concentrations or seaweed spouse concentrations were varied, and the growth growth numbers of seaweed spouses were compared one month later.

실시예 2-1Example 2-1

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 107 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 1g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a pestle bowl and pestle, and put into sterile seawater so that the spore concentration was 1.17 × 10 7 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 1 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterilized seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spouses growing by the seaweed was confirmed.

실시예 2-2Example 2-2

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 107 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 2g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a pestle bowl and pestle, and put into sterile seawater so that the spore concentration was 1.17 × 10 7 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 2 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-3Example 2-3

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 107 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 4g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the spore concentration was 1.17 × 10 7 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 4 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spouses growing by the seaweed was confirmed.

실시예 2-4Example 2-4

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 107 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 5g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a pestle bowl and pestle, and put into sterile seawater so that the spore concentration was 1.17 × 10 7 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 5 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-5Example 2-5

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 107 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 7.5g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a pestle bowl and pestle, and put into sterile seawater so that the spore concentration was 1.17 × 10 7 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 500 ml of the pulverized liquid and mixed with 7.5 g of alginate. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-6Example 2-6

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 107 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 10g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a pestle bowl and pestle, and put into sterile seawater so that the spore concentration was 1.17 × 10 7 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized solution was dissolved by mixing 10 g of alginate in 500 mL of the pulverized solution. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-7Example 2-7

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 106 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 1g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 6 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 1 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-8Example 2-8

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 106 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 2g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 6 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 2 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-9Example 2-9

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 106 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 Alginate 수용액의 제조는 분쇄액 500㎖에 알긴산염 4g을 녹여 혼합하였다. 혼합한 Alginate 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 6 cells/ml. To prepare the aqueous solution of alginate for coating the pulverized solution, 4 g of alginate was dissolved in 500 ml of the pulverized solution and mixed. After applying the mixed aqueous Alginate solution to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spouses growing by the seaweed was confirmed.

실시예 2-10Example 2-10

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 106 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 5g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 6 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 5 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-11Example 2-11

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 106 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 7.5g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 6 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 500 ml of the pulverized liquid and mixed with 7.5 g of alginate. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-12Example 2-12

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 106 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 10g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater to obtain a spore concentration of 1.17 × 106 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized solution was dissolved by mixing 10 g of alginate in 500 mL of the pulverized solution. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spouses growing by the seaweed was confirmed.

실시예 2-13Example 2-13

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 105 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 1g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 5 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 1 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-14Example 2-14

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 105 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 2g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 5 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 2 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-15Example 2-15

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 105 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 4g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 5 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in a mixture of 4 g of alginate in 500 ml of the pulverized liquid. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-16Example 2-16

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 105 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 5g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 5 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved in 5 g of alginate in 500 ml of the pulverized liquid and mixed. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-17Example 2-17

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 105 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 7.5g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 5 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized liquid was dissolved and mixed 7.5 g of alginate in 500 ml of the pulverized liquid. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

실시예 2-18Example 2-18

휴면 상태로 보존된 미역 배우체 덩어리를 배양액에서 분리하여 막자 사발과 막자를 이용해 분쇄하고 멸균 해수에 넣어 배우체 농도가 1.17 × 105 cells/㎖가 되도록 하였다. 분쇄액의 피막화를 위한 알지네이트(Alginate) 수용액의 제조는 분쇄액 500㎖에 알긴산염 10g을 녹여 혼합하였다. 혼합한 알지네이트(Alginate) 수용액을 해양 구조물에 도포 후 멸균해수 100㎖에 염화칼슘 3g을 녹여 제조한 경화제로 경화 및 침지하여 1 개월 후 미역 배우체의 착생 성장 개체 수를 확인하였다.The clump of seaweed spores preserved in the dormant state was separated from the culture medium, crushed using a mortar and pestle, and put into sterile seawater so that the concentration of the spores was 1.17 × 10 5 cells/ml. Preparation of an alginate (Alginate) aqueous solution for the coating of the pulverized solution was dissolved by mixing 10 g of alginate in 500 mL of the pulverized solution. After the mixed aqueous alginate solution was applied to a marine structure, 3 g of calcium chloride was dissolved in 100 ml of sterile seawater to cure and immerse with a curing agent prepared, and after 1 month, the number of seaweed spores was confirmed.

표 3은 알지네이트(Alginate) 수용액의 농도 또는 미역 배우체 수에 따른 착생 성장수의 비교한 결과이다. Table 3 is a comparison result of the number of growth growth according to the concentration of the alginate (Alginate) aqueous solution or the number of seaweed spouses.

구분division Alginate 수용액
(g/100㎖)Alginate aqueous solution
(g/100ml) 염화칼슘 경화제
(g/100㎖)Calcium chloride curing agent
(g/100ml) 배우체 수
(cells/㎖)Number of spouses
(cells/ml) 착생 성장수
(cells/㎠)Growth number
(cells/㎠) 실시예 2-1Example 2-1 0.20.2 33 1.17 × 107 1.17 × 10 7 0.05 ± 0.020.05 ± 0.02 실시예 2-2Example 2-2 0.40.4 33 1.17 × 107 1.17 × 10 7 0.11 ± 0.040.11 ± 0.04 실시예 2-3Example 2-3 0.80.8 33 1.17 × 101.17 × 10 77 1.82 ± 0.321.82 ± 0.32 실시예 2-4Example 2-4 1One 33 1.17 × 107 1.17 × 10 7 1.36 ± 0.331.36 ± 0.33 실시예 2-5Example 2-5 1.51.5 33 1.17 × 107 1.17 × 10 7 1.05 ± 0.091.05 ± 0.09 실시예 2-6Example 2-6 22 33 1.17 × 107 1.17 × 10 7 0.74 ± 0.100.74 ± 0.10 실시예 2-7Example 2-7 0.20.2 33 1.17 × 106 1.17 × 10 6 0.03 ± 0.010.03 ± 0.01 실시예 2-8Example 2-8 0.40.4 33 1.17 × 106 1.17 × 10 6 0.05 ± 0.010.05 ± 0.01 실시예 2-9Example 2-9 0.80.8 33 1.17 × 101.17 × 10 66 2.47 ± 0.442.47 ± 0.44 실시예 2-10Example 2-10 1One 33 1.17 × 106 1.17 × 10 6 1.90 ± 0.311.90 ± 0.31 실시예 2-11Example 2-11 1.51.5 33 1.17 × 106 1.17 × 10 6 1.64 ± 0.401.64 ± 0.40 실시예 2-12Example 2-12 22 33 1.17 × 106 1.17 × 10 6 1.19 ± 0.251.19 ± 0.25 실시예 2-13Example 2-13 0.20.2 33 1.17 × 105 1.17 × 10 5 -- 실시예 2-14Example 2-14 0.40.4 33 1.17 × 105 1.17 × 10 5 0.02 ± 0.010.02 ± 0.01 실시예 2-15Example 2-15 0.80.8 33 1.17 × 101.17 × 10 55 1.62 ± 0.391.62 ± 0.39 실시예 2-16Example 2-16 1One 33 1.17 × 105 1.17 × 10 5 1.33 ± 0.231.33 ± 0.23 실시예 2-17Example 2-17 1.51.5 33 1.17 × 105 1.17 × 10 5 0.82 ± 0.070.82 ± 0.07 실시예 2-18Example 2-18 22 33 1.17 × 105 1.17 × 10 5 0.55 ± 0.110.55 ± 0.11

표 3에서 알지네이트 염의 농도를 0.2 내지 2g/100㎖로 달리하여 착생성장 수를 비교 분석한 결과, 미역 배우체를 1.17 × 105 내지 1.17 × 107 cells/㎖의 농도로 포함하는 배양액을 이용한 경우 모두 알지네이트의 농도가 0.8g/100㎖에서 착생 성장수가 1.62 ± 0.32 내지 2.47 ± 0.44 cells/㎠으로 가장 우수함을 확인하였다. 이중 1.17 × 106 cells 농도의 미역 배우체를 이용한 경우 착생 성장수가 2.47 ± 0.44 cells/㎠로 가장 높게 나타났다. In Table 3, when the concentration of alginate salt was changed to 0.2 to 2 g/100 ml, and the number of engraftment growth was compared and analyzed, all the cases of using a culture solution containing seaweed spouses at a concentration of 1.17 × 10 5 to 1.17 × 10 7 cells/ml were used. It was confirmed that the concentration of alginate was the best at 0.8g/100ml, with the number of growths growing from 1.62 ± 0.32 to 2.47 ± 0.44 cells/cm 2. Among them, when seaweed spouses with a concentration of 1.17 × 10 6 cells were used, the growth number of seedlings was highest at 2.47 ± 0.44 cells/㎠.

반면, 알지네이트 수용액의 농도 0.4g/100㎖ 이하에서는 착생 성장수가 0.11 ± 0.04 cells/㎠로 측정되어 낮아 알지네이트 수용액을 0.4g/100㎖ 이하로 이용할 경우, 착생 개선 효과가 낮은 것을 확인하였다. 또한, 알지네이트의 농도가 1/100㎖ 에서 2/100㎖로 증가할수록 착생 성장수가 감소하는 경향을 나타냈으며, 동일한 미역 배우체 배양액 농도에서 비교하였을 때, 알지네이트 수용액의 농도가 2/100㎖일 때 효과가 가장 우수한 0.8g/100㎖의 비교하여 착생 성장수가 절반 수준(1.19 내지 0.55 cells/㎠)으로 감소하는 것을 확인하였다. On the other hand, when the concentration of the alginate aqueous solution was 0.4 g/100 ml or less, the number of growth of growth was measured to be 0.11 ± 0.04 cells/cm 2, and it was confirmed that when the alginate aqueous solution was used at 0.4 g/100 ml or less, the effect of improving the growth was low. In addition, as the concentration of alginate increased from 1/100 ml to 2/100 ml, the number of growth growths tended to decrease, and when compared at the same concentration of seaweed sperm culture, the effect of alginate aqueous solution concentration was 2/100 ml Compared with the most excellent 0.8g / 100ml it was confirmed that the number of growth growth is reduced to half the level (1.19 to 0.55 cells / ㎠).

Claims (4)

미역(Undaria pinnatifida) 배우체를 25 내지 27℃의 온도조건에서 1000 내지 2200 lx의 조도 조건으로 배양하여 휴면상태로 유지시키는 단계를 포함하는, 미역 배우체의 대량생산 방법.A method for mass production of seaweed spores, comprising culturing seaweed ( Undaria pinnatifida ) spores at a temperature of 25 to 27°C under an illumination condition of 1000 to 2200 lx to maintain a dormant state. 제 1항에 있어서,
상기 미역 배우체는 광주기 조건 12:12(명기: 암기)로 배양되는 것을 특징으로 하는, 미역 배우체의 대량 생산방법.According to claim 1,
The seaweed spouse is characterized by being cultured under Gwangju period condition 12:12 (memory: memorization), a method for mass production of seaweed spouse. 제 1항의 생산방법으로 생산한 휴면상태의 미역 배우체를 105 내지 107 cells/㎖의 농도로 포함하는 배양액을 제조하는 단계;
상기 배양액 100 중량부에 대하여 알지네이트를 0.6 내지 1.8 중량부로 혼합하여 혼합물을 제조하는 단계; 및
상기 혼합물에 다가 금속염을 첨가하여 경화시켜 피막화된 미역 배우체를 제조하는 단계를 포함하는, 미역 배우체의 착생 개선 방법.Preparing a culture medium containing a dormant seaweed spouse produced in the production method of claim 1 in a concentration of 10 5 to 10 7 cells / ㎖;
Preparing a mixture by mixing alginate in an amount of 0.6 to 1.8 parts by weight based on 100 parts by weight of the culture solution; And
And adding a polyvalent metal salt to the mixture to cure to prepare a coated seaweed spore. 제 3항에 있어서, 상기 다가 금속염은 염화칼슘, 염화마그네슘, 염화철(2가), 염화철(3가), 황산칼슘, 황산마그네슘, 황산철(2가) 및 황산철(3가) 로 이루어진 군에서 선택되는 1종 이상인 것을 특징으로 하는 미역 배우체의 착생 개선 방법.According to claim 3, The polyvalent metal salt is in the group consisting of calcium chloride, magnesium chloride, iron chloride (divalent), iron chloride (trivalent), calcium sulfate, magnesium sulfate, iron sulfate (divalent) and iron sulfate (trivalent) Method for improving the growth of seaweed spouses, characterized in that at least one selected.
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