초록 ▼

Ⅴ. 연구개발결과
● 팔레놉시스/과수류 대량생산 체계확립 및 위탁생산 시설 구축
- 사과왜성대목의 대량생산 체계확립 및 규격 포트묘 94,146주 보급체계구축
- 팔레놉시스 대량생산 및 규격 포트묘 667,103주 보급 체계구축
- 블루베리 대량생산 및 규격 플러그묘 325,712주 보급 체계구축
- 민간 위탁 생산시설 구축(난류, 과수류, 숙근류 순화실 1개소, 조직배양실 1개소구축)
● 팔레놉시스 릴레이 (relation ship)배양 체계확립 및 국내육성 품종 대량생산
- 부정아 방식으로

Ⅴ. 연구개발결과
● 팔레놉시스/과수류 대량생산 체계확립 및 위탁생산 시설 구축
- 사과왜성대목의 대량생산 체계확립 및 규격 포트묘 94,146주 보급체계구축
- 팔레놉시스 대량생산 및 규격 포트묘 667,103주 보급 체계구축
- 블루베리 대량생산 및 규격 플러그묘 325,712주 보급 체계구축
- 민간 위탁 생산시설 구축(난류, 과수류, 숙근류 순화실 1개소, 조직배양실 1개소구축)
● 팔레놉시스 릴레이 (relation ship)배양 체계확립 및 국내육성 품종 대량생산
- 부정아 방식으로 증식된 팔레놉시스 →발근, 정식 릴레이 배양 309,170주 생산체계 확립
- 릴레이 배양과정 중 발근효율 증대→발근배지 선발
- 국내육성 품종 대량생산 및 920,500주 보급체계 구축
● 조직배양에 의한 화훼류 및 관엽류의 대량생산
- 협동 3기관으로부터 관엽 무병주 및 초대배양체 분양→릴레이 배양에 의한 대량생산
- 관엽류, 고무나무, 고구마, 칼라디움 등 222,000주 생산보급
- 관엽, 고구마, 화훼류 대량생산 및 배지구명
● 조직배양실의 생력화 및 순화실 환경제어 프로그램 개발
- 조직배양묘 블루베리, 고구마, 감자 순화환경 구명
. 블루베리 순화 후 육묘에 적합한 배양액의 EC 1.2 dS·cm^-1에서 초장 4.3 cm 와 엽수 21.5개로 가장 높았고, pH 4.0으로 조절하고 NH₄SO₄ 1mM 를 첨가하였을 때 대조구(EC 1.0, pH 4.0) 에 비해 초기 육묘용 생육에 효과적이었다.
. 고구마 LED 광파장(white: 400∼450nm, 500∼780 nm, red: 700∼780nm, blue:
400∼480nm, R:B=3:7, future green(FG): 400∼800nm)에 따라 재배하였을 때 생체중 및 엽수가 많고 초장이 짧은 R:B가 3:7로 혼합된 LED가 적합하였다.
. 가공용 신품종 ‘새봉’의 조직배양묘를 EC 수준 (0.6, 1.0, 1.4, 1.8 dS·m^-1)을 달리 순화하였을 때, 순화 6일째 생존율은 EC 0.6∼1.0 처리에서 100% 였으며 EC 1.4∼1.8 처리에서는 0% 로 순화를 위한 적정 농도는 EC 0.6∼1.0 dS·m^-1 였다.
- 순화실 환경제어 프로그램 개발
. 조직배양 생산이력 추적관리 프로그램 개발
. 육묘 생산이력 관리 프로그램 개발
. 육묘 환경모니터링 프로그램 개발
- 순화용 온실에서 원예작물의 Microponic system 개발 및 실증
. 조직배양묘 순화용 온실환결 계측
. Microponic system 설치운영
. 팔레놉시스, 블루베리, 사과왜성대목 대량순화 실증및검증
● 조직배양식물의 바이러스 무병모주 양성
- 바이러스 무병모주 양성 및 분양(국화, 고구마, 나리, 칼라)
- 초대배양체 기내 확립 및 분양(감자, 사과, 배, 양앵두 대목)
- 바이러스 검증 총 137종의 화훼, 과수작물 바이러스 검증
● 우량 무병묘 생산관리 기술개발
- 블루베리 조직배양묘 기내배양시 오염율 경감방법 개발
. 배지 내 항균제 PPM^TM 1㎖/L 처리로 식물체 생육 저해(약해) 없이 배양 오염율 52∼59% 감소
- 블루베리 기내 배양묘 변이 발생 및 유전적 안정성 확인
. 블루베리 우량 배양묘 안전 생산을 위한 계대배양 적정 횟수는 10회
. 배양묘 생산효율 및 경제성 고려 시 계대배양 15회 이내 권장
- 생산단계별 배양묘 바이러스 검정 및 포장 실증을 통한 국산 무병묘 품질검증
. 기내배양묘/기외순화묘/포장정식묘 단계별 바이러스 검정
. 삽목묘 대비 배양묘 주축지 및 신초수 증가로 주당 과실생산량 20% 증가
● 세부과제별 성과요약
■ 민간 위탁 대량생산시설 구축 및 국내 육성품종 대량생산
○ 세부1
팔레놉시스/과수류 대량생산 체계확립 및 위탁생산 시설 구축
- 팔레놉시스 대량생산 및 규격 포트묘 671,313주 보급
- 블루베리 대량생산 및 규격 플러그묘 325,712주 보급
- 사과왜성대목의 대량생산 체계확립 및 규격 포트묘 94,146주 보급
- 민간 위탁 생산시설 구축(난류, 과수류, 숙근류 순화체계 확립)
○ 협동1
팔레놉시스 릴레이 (relation ship)배양 체계확립 및 국내육성 품종 대량생산
- 부정아 방식으로 증식된 팔레놉시스 →발근, 정식 릴레이 배양 309,170주 생산체계 확립
- 릴레이 배양과정 중 발근효율 증대→발근배지 선발
- 국내육성 품종 대량생산 및 920,500주 보급
■ 건전 우량묘 생산체계 확립을 위한 시스템 개발
○ 협동2
조직배양실의 생력화 및 순화실 환경제어 프로그램 개발
- 조직배양묘 블루베리, 고구마, 감자 순화환경 구명
- 순화실 환경제어 프로그램 개발
. 조직배양 생산이력 추적관리 프로그램 개발
. 육묘 생산이력 관리 프로그램 개발
. 육묘 환경모니터링 프로그램 개발
- 순화용 온실에서 원예작물의 Microponic system 개발 및 실증
. 조직배양묘 순화용 온실환결 계측
. Microponic system 설치운영
. 팔레놉시스, 블루베리, 사과왜성대목 대량순화 실증 및 검증
○ 협동3
조직배양식물의 바이러스 무병모주 양성
- 바이러스 무병모주 양성 및 분양(국화, 고구마, 나리, 칼라)
- 초대배양체 기내 확립 및 분양(감자, 사과, 배, 양앵두 대목)
- 바이러스 검증 총 137종의 화훼, 과수작물 바이러스 검증
- 화훼, 고구마 대량번식 1,571,000주 보급
위탁 3-1) 조직배양에 의한 관엽류의 대량생산
- 협동 3기관으로부터 관엽 무병주 및 초대배양체 분양→릴레이 배양에 의한 대량생산
- 관엽류, 고무나무, 고구마, 칼라디움 등 222,000주 생산보급
- 관엽, 고구마, 화훼류 대량생산 및 배지구명
○ 협동4
우량 무병묘 생산관리 기술개발
- 블루베리 조직배양묘 기내배양시 오염율 경감방법 개발
- 블루베리 기내 배양묘 변이 발생 및 유전적 안정성 확인
- 생산단계별 배양묘 바이러스 검정 및 포장 실증을 통한 국산 무병묘 품질 검증

Abstract ▼

We carried out establishment of contract manufacturing through plant tissue culture of superior seedlings. Including phalaenopsis, blueberry and apple draft rootstock M9 and M26, plantlets were produced and supplied by plant tissue culture. The results are follows.
1. Phalaenopsis/fruit crops est

We carried out establishment of contract manufacturing through plant tissue culture of superior seedlings. Including phalaenopsis, blueberry and apple draft rootstock M9 and M26, plantlets were produced and supplied by plant tissue culture. The results are follows.
1. Phalaenopsis/fruit crops established the mass production system and contract manufacturing facilities building
- Built mass production system and establish automatic seedling production system, and we produced 94,146 clone seedlings of apple dwarf rootstocks
- Phalaenopsis mass-produced 667,103 clone seedlings used automatic seedling production system and tissue culture propagation method.
- Construction of blueberries mass production and supply system standard plug seedlings 325,712 clone seedlings
- Contracting production facilities construction(One acclimatization system and Tissue culture room )
2. Establishment of Phalaenopsis relationship culture system and mass production of domestic varieties
- Establishment in rooting system of adventitious propagation method and produced 309,170 clone seedlings.
- Select rooting medium during the culture process of relationship culture system
- 920,500 clone seedlings produced of domestic varieties.
3. Automation of tissue culture system and environmental control program development
○ Appropriate acclimatization environment for tissue culture seedling of blueberries
- The objective of this study were to establish a in vitro culture with hydroponic by determining optimum environments followed by optimum growth of blueberry nursery plant.
- In vitro blueberry nursery plants were transplanted of a hydroponic 106days. For FL, LED, White LED photosynthetic photon flux(PPF) levels of 80 and 150 μmol·m^-2·s^-1, electrical conductivity(EC) level of 0.8 dS·m^-1, pH levels of 5.0, substrates kinds of peatmoss, perlite, coconut coir, rockwool, polyphenol resin, PMLV(Peat:Perlite:Vermiculite=6:3:1), and PML(Peat:Perlite=5:5) and hydroponic kinds of aeroponics, DFT and NFT were provided to investigate the effects of the treatments on growth of nursery plant of the blueberry cuttings.
- Plant height decreased with increasing LED PPF. blueberry transplants grown under LED PPF of 150 μmol·m^-2·s^-1 exhibited noticeably compact shoot growth with increased leaves. Plantlets with shoot weight, plant height, root length, leaf length, leaf width and leaf number will be more suitable for transplanting to ex vitro condition.
- Optimum growth rate of the nursery plant blueberry was controlled to EC 0.8 dS·m^-1 with 5.0 for pH. Optimum light source, photosynthetic photon flux and substrate for the growth of the blueberry nursery plant in the NFT hydroponic were determined to LED, 150 μmol·m^-2·s^-1 for photosynthetic photon flux(PPF) and peatmoss for substrate. Appropriate acclimatization environment for raising seedlings of blueberries
- Plant height and number of leaves was the highest After blueberry acclimatization in EC 1.2dS·m^-1. adjusted to pH 4.0 and was effective for early seedling growth compared to the control (EC 1.0, pH 4.0) was added to the 1 mM NH4SO4.
○ Appropriate acclimatization environment for tissue culture seedling of sweet potato
- Sweet potato acclimatization rate was 100% after 4 weeks when the relative humidity keep more than RH 85%. there fore rootless acclimatization is effective. Acclimatization rate was highest when retaining 95% 1 day, 85% 2 days, 3 days to 75% and rooting was vigorous between 3 to 5 days. It requires maintenance of minimum 1 to 2 days in RH 95% to facilitate rooting.
- LED light wavelength (white: 400~450 nm, 500~780 nm, Red: 700~780nm, Blue:400~480nm, R: B = 3: 7, Future Green (FG): 400 to 800 nm) when grown in accordance with the live weight and number of leaves a lot shorter plant height R: B 3:7 mixed LED was suitable
○ Appropriate acclimatization environment for tissue culture seedling of potato
- It was intended to closely examine an effect that a change in the concentration of culture medium had on the potato(Solanum tuberosum L.) plantlet growth in the microponic system so as to mass-produce the virus-free plant of new variety ‘Saebong’ for potato processing.
- The adjusted concentration of potato culture medium was 0.2, 0.6, 1.0, 1.4,1.8, and 14.0 dS·m^-1. And potato seedling was cut into pieces of 1.5 cm in length, which included 2 growth points and leaves. And each was explanted in glass vial of 50 mL. And experiments were carried out twice for 18 days or 21days. Culture medium of 2ml was put in the container respectively. And 1 mL was added after 10 days. And in terms of cultivation environment, the experiment was carried out at the day length of 16 hours at the temperature of 23±1℃ under the white LED light of 40 μmol·m^-2·s^-1.
- The results showed that the survival rate of plantlet was 90% at 0.2 dS·m^-1, 100% at 0.6 dS·m^-1, 100% at 1.0 dS·m^-1. 0% at 1.4 dS·m^-1, 0% at 1.8 dS·m^-1. and 0% at 14.0 dS·m^-1 after 7 days. With regard to the explanted potato seedling, in case of the treatment where the electrical conductivity of culture medium was adjusted to 1.0 dS·m^-1, root developed 2 days after transplantation. And the plantlet vigorously grew into strong plant that had 7 leaves, length of 5cm, and fresh weight of 0.5 g after 18 days. In case of the treatment where the concentration of culture medium was adjusted to 0.6 dS·m^-1, the root plantlets developed 4 days after transplantation. And those grew into plant that had 7 leaves and fresh weight of 0.2 g after 21 days. Therefore, we found that it is effective to control potato culture medium by adjusting its electrical conductivity to 0.6~1.0 dS·m^-1 for the mass production of virus-free potato seedling in the microponic system.
○ Microphonic system development and demonstration of horticultural crops in the greenhouse for acclimatization
- Hydroponic differentiation and seedling cultivation takes place at the same time creating an environment control system was automated and available in two energy saving the nursery system available to lead agency. In addition, this system has been configured to also apply a variety of crops, as well as blueberries.
○ Development of environment control program for acclimatization
- Seedling production traceability programs and tissue culture production traceability management program was developed for tissue culture room energy savings. Environment control program has been developed by the Light, temperature, humidity measurement compensation. After registration program plans to transfer technology to the tissue cultrue enterprise.
4. The virus free stocks were established in floricultural crops, sweet potato, potato and fruit rootstocks.
○ The establishment of virus free stocks in floricultural crops, sweet potato and potato.
- The establishment of virus free stocks and in vitro mother plants : The twenty virus free stocks were established in chrysanthemums including "Dream Land", multiplicated on MS mediun with 2g/L activated charcoal, and supplied. The establishment of in vitro plantlets in Anthurium, Alocasia, and 2 Ficus species.
- The establishment of virus free stocks in sweet potato and potato : In six sweet potato cultivars, virus free stocks were establishmented, and multiplicated and supplied on MS medium with 0.1 mg/L NAA. The virus free stocks in three cultivar potato were establishmented and multiplicated on MS mediun without plant growth regulators.
○ The establishment of virus free stocks in floricultural crops, fruit three and strawberry.
- The virus free rootstocks in fruit tree and supply : The virus free rootstocks were establishmented in apple in M.9., M.26., pear rootstock "Baeyeun 3" and cherry "Super 6", and multiplicated.
- In the two straw berries including "Dae Wang", virus free mother plantlets were establishmented.
- Adequate media were founded in foliage crops and multiplicated especially in Aglaonema and Ficus.
- In vitro culture of crops by plant tissue culture : The twenty thousand plants in 15 sweet potato cultivars was produced and supplied. The twenty eight virus free cultivars were established and micropropagated in Lillium, calla and chrysanthemum.
○ The establishment of virus free stocks in floricultural crops, fruit tree and sweet potato
- The establishment of virus free stocks in chrysanthemum cultivar "Baekma" et al.
- The virus free mother plants were establishmented in six sweet potato cultivars, and 300 thousand plants were supplied.
- The virus free rootstocks in fruit trees were multiplicated and supplied.
5. In foliage plants, sweet potatos and fruit trees, Many plantlets were produced and supplied by plant tissue culture. The results are follows.
○ In the first year, the seventy thousand plantlets in Agreonema and Syngonium were multiplicated, and supplied. Agreonema was multiplicated on MS mediun with 0.5 mg/L TDZ and 0.2 mg/L NAA, and Syngonium on MS mediun with 3.0 mg/L 2ip and 0.5 mg/L NAA. Ficus was cultured on MS with 1.0 mg/L BA and 0.1 mg/L IAA, and Ficus 'Bahalensis' on MS with 2.0 mg/L BA and 0.5 mg/L NAA.
○ In the second years, adequate media were investigated in foliage crops prefered in the nation. The 5,000 plantlets in Alocasia were produced by tissue culture, and supplied, 50,000 ones in Agreonema, and 20,000 plantlets in Ficus. Including cultivar "Dahomi" 40,000 plantlets of sweet potato were produced by tissue culture.
○ In the third years, 50,000 plantlets of sweet potato were produced, 12,000 plantlets in Caladium "White Ball", and 20,000 ones in Ficus. Apple rootstock M.9., and M.26. are producing by tissue culture.
6. Development of technology for the mass propagaion of virus-free nursery stock
○ Interest and great demand for blueberry (Vaccinium corymbosum) have increased, as V. corymbosum is now one of the most economically important crops in Korea. It is expected that blueberry production and the area planted for cultivation will increase consistently in the years ahead because of high profitability and the consumer's demand for healthy ingredients. Effective mass production of blueberry is urgently needed for commercial cultivation establishment, but a main limitation is lack of a propagation system that produces a disease-free plant material for commercial plantation. A large amount of research has focused entirely on developing tissue culture techniques for blueberry propagation.
○ However, controlling fungal and bacterial contamination of woody plant material is extremely difficult. Our study was conducted to investigate the effect of biocide addition during the in vitro culture of blueberry on plantlet growth and contamination occurrence. Four biocides, including Plant Preservative Mixture (PPM^TM), Vancomycin, Nystatin and Penicillin G, were used in varying concentrations during the in vitro propagation of blueberry. When nystatin was added into the medium at low concentrations, the overall growth of blueberry plantlets was retarded. Addition of vancomycin and penicillin G in high concentrations decreased contamination but induced plantlet mortality. On the other hand, when 1㎖/L PPM^TM was added, the growth characteristics of blueberry plantlets did not significantly differ from non-treatment (control), and the contamination occurrence rate was very low. From these results, we found that the addition of the appropriate biocide could provide an effective method to reduce contamination in the culture process, thereby raising in vitro production efficiency.
○ We surveyed the growth differences of in vitro blueberry plantlets propagated from apical meristem and leaf explant. The length of plantlets cultured from apical meristem increased by 18~31%, but the number of new shoot increased by 1.5~1.8 times in the plantlets cultured from leaf explant. The changes in blueberry shoot proliferation were investigated after repeated in vitro subculture. The shoot death rate increased by 2.8 times, and the morphological variation of leaf was also detected, furthermore, ex vitro acclimatization rate decreased by 12% after fifteenth subculture, in comparison with the results from fifth and tenth subculture. Therefore, we found that the vigorous and healthy blueberry plantlets could be produced within the properly repeated subculture.
○ We checked the possibilities of virus detection in the various propagation stages from in vitro culture vessel to field by ELISA. All samples showed the negative responses when the detection tests of 4 viruses (BlScV, BShV, BLMoV, BSsV) were conducted.
○ Field performance of highbush blueberries propagated by cuttings and tissue culture was also accomplished for 3 years. There were no significances in flowering and fruit characteristics, tree height as well as the harvesting time. But the number of main branch and new shoot increased significantly by 50% and the production yield of fruit also increased by 20~30%. Therefore, it seemed that the cultivation of blueberry plants propagated by tissue culture could induce the high yield if the proper pruning and fertilization were applied.