발간등록번호 11-1543000-001499-01 배발생세포공정배양및토양정밀양구를통한 튤립개화구대량생산기술개발 Mass production of tulip bulbs through the embryogenic cell technologies and refined soil cultivation 단국대학교 목포대학교 농림축산식품부
주의 1. 이보고서는농림축산식품부에서시행한농생명산업기술개발사업의연구보고서입니다. 2. 이보고서내용을발표하는때에는반드시농림축산식품부에서시행한농생명산업기술개 발사업의연구결과임을밝혀야합니다. 3. 국가과학기술기밀유지에필요한내용은대외적으로발표또는공개하여서는아니됩니다.
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보고서요약서 과제고유번호 111072-05- 2-SB010 해당단계 연구기간 11.09.23 ~ 16.09.22 단계구분 (5)/ (5) 중사업명 연구사업명 세부사업명 연구과제명 대과제명 세부과제명 배발생세포공정배양및토양정밀양구를통한튤립개화구대량생산기술개발 해당단계 참여 연구원수 총 : 7 명 내부 : 명 외부 : 명 해당단계 연구개발비 정부 :100,000 천원 민간 :25,000 천원 계 :125,000 천원 연구책임자 총연구기간 총 : 명 정부 :500,000 천원 참여 내부 : 명 총연구개발비 민간 :125,000 천원 연구원수 외부 : 명 계 :625,000 천원 연구기관명및 소속부서명 단국대학교천안캠퍼스산학협력단 목포대학교산학협력단 참여기업명 한국화훼구근산업영농조합법인 위탁연구 연구기관명 : 연구책임자 : 요약 ( 연구개발성과를중심으로개조식으로작성하되, 500 자이내 로작성합니다 ) 보고서면수 - 3 -
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Purpose& Contents Results Expected Contribution D-02 In vitro bulblet production methods were developed using somatic embryogenic cell culture technologies for major tulip cultivars. Embryogenic cell lines were induced and proliferated in small bioreactors, and then regenerated and cultured to form bulblets. To optimize tulip bulb production practices in Chonnam province, soil characteristics of the region were investigated. And cultivation formats for tulip bulb production were established by investigating the most effective irrigation, fertilization and cultivation protocols. Tulip bulbs multiply 2.5 times a year, the lowest rate among cultivation crops. Vegetative methods are not available and tissue culture method is hardly practiced due to its recalcitrant propagation properties. We developed in vitro techniques of embryogenic cell induction and multiplication for many cultivars of tulip. And the methods were optimized for large scale cultures using bioreactors and bulblets were produced following regeneration on solid medium. Tulip cultivations in soil were also optimized for bulb production in Chonnam province. Development of embryogenic cell culture technology for tulip bulblet production. - Embryogenic cells were induced in 15 commercial cultivars of tulip. - The cell lines were proliferated in 5-L bioreactors. - Browning problems during the liquid cultures were overcome by optimizing the cultures. - Embryogenic cells were transferred to solid media and bulblets were formed through embryogenesis. - The bulblet production procedures were optimized for mass production of major tulip cultivars. Establishment of tulip soil cultivation protocols for flower bulb production - Physicochemical characteristics of soils in Chonnam province were characterized. - Bulb planting depth, space and period were optimized. - Fertilization practices for bulb enlargement - Irrigation practices for each step of tulip cultivation Cell culture technologies would produce virus-free bulblets of tulips which were cultivated in greenhouse and soil to produce flower bulbs either in domestic or foreign area such as Da lat, Vietnam. Tulip cultivation protocols were established in Chonnam province and high quality bulbs can be produced for farmers at lower prices. The knowhow is being transferred to a corporate for industrialization. This will create new opportunities for the domestic and global utilization of tulip bulbs. Keywords Tulip embryogenesis cell culture bulblet flower bulb - 6 -
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D-05 Cultivars Plant Growth Regulators Total no of explant Brown -ing Death Callus Total number of callus Callus -ing rate (%) Dynasty 2,4-D BA 865 - - 145 18 - Golden apeldon Golden parade Hollandia Ile de france Louvre Mark Voort Callus weight (mg) 2,4-D BA 282 - - 69 24 - Picloram BA 60 - - 40 66 - Dicamba BA 24 - - 24 100-2.4-D Picloram Dicamba BA 34 18 5 33 97 260 kenetin 43 19 15 21 49 410 2ip 41 7 3 39 95 670 BA 53 3 3 50 94 640 kenetin 48 25 21 24 50 640 2ip 52 10 13 44 85 570 BA 39 8 6 26 67 380 kenetin 30 8 8 19 63 780 2ip 43 14 14 21 49 800 2,4-D BA 189 - - 89 47 - Picroram BA 101 101-91 90 - Dicamba BA 9 0-1 11-2.4-D Picloram Dicamba BA 90 14 17 23 26 274 kenetin 90 24 29 8 9 154 2ip 90 12 7 40 44 76 BA 90 8 13 29 32 328 kenetin 90 5 2 27 30 439 2ip 87 6 7 13 15 222 BA 102 14 20 24 23 315 kenetin 88 15 11 15 17 477 2ip 82 22 36 4 5 193 2,4-D BA 45 - - 13 29 - Picroram BA 8 - - 6 75 - Picroram BA 93 - - 22 24-2,4-D BA 97 - - 67 70 - - 26 -
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D-05 튤립구근 RT-PCR 진단 LSV * CMV TBV TRV ArMV LMoV 전자현미경 1 음성음성음성음성음성음성바이러스입자없음 2 음성음성음성음성음성음성바이러스입자없음 - 39 -
D-05 * LSV: Lily symptomless virus( 백합무병징바이러스 ) CMV: Cucumber mosaic virus( 오이모자이크바이러스 ) TBV: Tulip breaking virus( 튤립브레이킹바이러스 ) TRV: Tobacco rattle virus( 담배얼룩바이러스 ) ArMV: Arabis mosaic virus( 아라비스모자이크바이러스 )-검역관리바이러스 LMoV: Lily mottle virus( 백합무병징바이러스 ) - 40 -
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D-05 시료품종 : 튤립구근 Ile de France 와 Come back 품종각 3점 의뢰내용 : 국립원예특작과학원원예특작환경과에튤립구근의바이러스진단을의뢰 2) 진단결과 및 RT-PCR을이용한유전자진단결과의뢰한튤립 2품종각각 3점에서바이러스가검출되지않았음 < 튤립바이러스진단결과 > 튤립구근 RT-PCR 진단 LSV * CMV TBV TRV ArMV LMoV 전자현미경 1 음성음성음성음성음성음성바이러스입자없음 2 음성음성음성음성음성음성바이러스입자없음 * LSV: Lily symptomless virus( 백합무병징바이러스 ) CMV: Cucumber mosaic virus( 오이모자이크바이러스 ) TBV: Tulip breaking virus( 튤립브레이킹바이러스 ) TRV: Tobacco rattle virus( 담배얼룩바이러스 ) ArMV: Arabis mosaic virus( 아라비스모자이크바이러스 )-검역관리바이러스 LMoV: Lily mottle virus( 백합무병징바이러스 ) - 116 -
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D-08 코드번호 D-09 코드번호 D-10 구입기관연구시설 / 연구장비명 규격 ( 모델명 ) 수량 구입연월일 구입가격 ( 천원 ) 구입처 ( 전화번호 ) 비고 ( 설치장소 ) NTIS 장비 등록번호 코드번호 D-11-121 -
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