초록 ▼

Ⅴ. 연구개발 결과
○ 씨감자 생산기술 및 생리적 활력 조절기술 개발
- 무병씨감자를 년중 생산할 수 있는 Pilot 시스템 개발
․온도와 광 등 식물생육환경을 인공적으로 조절하는 Pilot 시스템 개발
․영양공급, 환경조절 기술은 다양한 생태 환경 조건에서도 연중생산 가능한 최상위단계의 씨감자를 생산기술 개발
- 수출용 이동형 씨감자 식물공장 개발
․수출 대상국의 다양한 기후와 환경에 모두 적용이 가능한 인공환경조절 이동형 씨감자 식물공장 개발
․초미세분무방식(FOG 시스템) 새로운 수경재배

Ⅴ. 연구개발 결과
○ 씨감자 생산기술 및 생리적 활력 조절기술 개발
- 무병씨감자를 년중 생산할 수 있는 Pilot 시스템 개발
․온도와 광 등 식물생육환경을 인공적으로 조절하는 Pilot 시스템 개발
․영양공급, 환경조절 기술은 다양한 생태 환경 조건에서도 연중생산 가능한 최상위단계의 씨감자를 생산기술 개발
- 수출용 이동형 씨감자 식물공장 개발
․수출 대상국의 다양한 기후와 환경에 모두 적용이 가능한 인공환경조절 이동형 씨감자 식물공장 개발
․초미세분무방식(FOG 시스템) 새로운 수경재배 시스템 개발
- 수경재배 미니씨감자를 이용한 생력화 파종방법 구명
․미니씨감자 종구크기별 씨앗테이프 이용한 파종 노력비 절감 및 2립 파종시 수량성 증대
- 수경재배산 미니씨감자의 수량성 증대 효과 구명
․괴경형성처리를 정식 40일경에 pH4.0에서 2일 처리해주고 ‘대지’ 경삽묘는 봄재배는 경삽 25∼30일묘, 가을재배는 경삽 20∼30일묘가 적합
- 베트남 ‘솔라라’ 품종 묘소질 구명 및 저장기간 구명
․봄재배는 경삽 30∼35일묘, 가을재배는 경삽 35∼40일묘가 수량성 양호하였고 4℃에서 7개월 저장 가능
- 국외 품종 현지 실증시험으로 씨감자 우수성 입증 및 종서크기 구명
․베트남 ‘솔라라’, 카자흐스탄 ‘산테’ 품종 현지 실증재배로 480㎏ 생산 및 5∼10g에서 2∼3립 파종하는 것이 유리
- 수출 대상국 기후환경 분석
․중국, 몽골, 베트남, 우즈벡의 온도, 강수량, 일조시간 등 수출 대상국 환경 분석
․서령조절 및 저장기술 적용방법 확립
․수출 대상국 저장고 현황 및 저장 상황 조사
- 수출 대상국 작부체계 설정 및 씨감자 서령조절 기술 확립
․ 맹아억제제와 광처리를 통한 서령조절 효과 탐색

○ 수출용 씨감자 대량 생산 파일롯 시스템 개발
- 인공 환경 조절 기술 개발 : test room 설계 및 제작, 인공 환경 제어 장치 개발 및 프로그램 개발
- 조직배양용기 개발 : 배양용기 문제점 개선 및 금형 개발, 배양용기 성능 test
- 소규모 pilot plant system 개발
· 컨테이너형 조직배양실 시설 설계 및 제작
· 씨감자 생산 방법에 따른 pilot plant 설계 및 제작
· 국내 시제품 test 및 Target Market 현지 test
- 대규모 씨감자 생산 pilot plant system 개발
· Target Market 현지 조사 및 plant 설계

○ 병해충 저항성 평가 및 관리기술 개발
- 수출대상국의 문제병해충 선정 : 5개 주요 수출대상국에 대한 문제병해충 선정 완료
- 감자 주요 병해충 저항성 평가 시스템 개발
· 감자 역병, 풋마름병 및 더뎅이병에 대한 새로운 저항성 평가법 개발
· 평가방법을 이용한 유전자원 및 육성계통의 저항성 평가를 실시
· 주요 해충 4종에 대한 저항성 평가법 개발 완료
- 수출대상국의 해충에 대한 약제 저항성 평가 및 실태조사
· 베트남, 몽골, 카자흐스탄에 대한 실태조사 완료
· 살충제 저항성 진단 키트 및 콜로라도감자잎벌레 저항성 평가법 개발
- 감자 바이러스 진다키트 대량생산을 위한 항체 생산기술 개발
· 재조합 항체 선발 및 기내 생산 시스템 구축
· 바이러스 진단 키트 생산을 위한 벡터 개발
· ELISA 및 Immunostrip 시제품 생산

(출처 : 요약문 p.3)

Abstract ▼

The purpose of this project was to develop a new hydroponic system for seed tubers and nutrient management techniques for controlling haulm growth, and to determine if the effects differ among cultivars and growing seasons. Considering the difficulties for producing seed potatoes (Solanum tuberosum)

The purpose of this project was to develop a new hydroponic system for seed tubers and nutrient management techniques for controlling haulm growth, and to determine if the effects differ among cultivars and growing seasons. Considering the difficulties for producing seed potatoes (Solanum tuberosum) in target countries, techniques for use of rooted sprouts from small tubers and production of basic seed under environment-controlled conditions were included in the project.

Hydroponic culture of potatoes is a technology for production of seed tubers in nutrient solutions without soil. Till now, some pieces of information have been published on potato growth in hydroponic systems; water culture, modified water culture, NFT, aeroponics and substrate cultivation system using perlite and clay balls. Most of the hydroponics have limitations for growing potatoes in areas or countries where water sources are contaminated and not enough for potato production. In the present project, a water-saving hydroponics with fine mist (so called FOG) was developed and compared with the conventional aeroponics, for the production of potato seed tubers. Shoot fresh weight was significantly decreased when potato plants were grown in the FOG system compared to aeroponics. This decrease of shoot growth in the FOG system was accompanied by a decrease of seed tuber yields in the first two years (2013-2014) of the study. However, with following development of optimum misting conditions of the FOG system, there were no significant differences in days to tuberization and tuber yields between the two hydroponic systems. It could be concluded that the two systems produced a similar number of tubers, whereas FOG system could save water consumptions by decreasing the growth of fresh shoots than aeroponics.

In potato hydroponics, tuberization is often delayed because of increased growth and an extended vegetative cycle. Delayed tuberization is primarily caused by external conditions, including abiotic factors such as growing season, growing system, temperature, nutrients, pH, and water stress, as well as biotic factors like growth stage, hormones, and genotypes. The purpose of this study was to evaluate the growth and yield response of aeroponically grown potato plants to change in the nitrate (NO₃^-)/potassium (K) ratio in nutrient solution, and to determine the intervals of misting nutrient solution. Seasonal experiments during spring and autumn were conducted using both Korean and target countries’ potatoes, cvs. Solara and Sante. Characteristics of nutrient uptake as affected by NO3/K ratio in nutrient solution were analyzed. During autumn, total chlorophyll contents and photosynthetic rate in leaves decreased significantly with reduced NO3/K ratio. Shoot growth and days to tuberization were lower and shorter with the reduced NO3/K ratio in both spring and autumn seasons. In spite of the effect on shoot growth and tuberization, tuber growth was not affected by NO3/K ratio. Tuber numbers tended to increase with increased ratios of NO3/K during the autumn season, but no difference was noted during the spring season. The intervals between nutrient solution misting decreased the growth of shoot and tubers, when potato plants were exposed the misting every 30 minutes, although the significance were different depending on cultivars and seasons.

This research was conducted to inform excellence of Korean seed potatoes produced by hydroponics mode to Southeast Asia and Northern areas and to examine the possibility of export. Hydroponic produced small potatoes (3∼5g) used seed tape to sow 2 tubers by machine and as a result, 75% of labor charge was reduced. At demonstrative cultivation in Vietnam, sowing 2∼3 tubers around 3∼10g increased the cultivated amount and proved to be useful in utilizing seed potatoes. In spray hydroponic culture, treating root with pH 4.0 for two days around 40 days after planting helped increase the amount of seed potato tubers. For seedling used in planting 'Daeji' cultivar, 25∼30th day stem cutting seedling for spring cultivation, and 20∼30th day stem cutting seedling was favorable respectively.

Also for Vietnamese 'Solala' cultivar seedling used for planting, 30∼35th day stem cutting seedling for spring cultivation, and 35∼40th day stem cutting seedling was favorable for its amount. Also in storage examination, it did not have any problem in its storage and germination in 4℃ for 7 months. Foreign seed potatoes produced by Korean hydroponic culture showed excellent results in Vietnam and Kazakhstan when demonstratively cultivated locally. Vietnamese 'Solala' and Kazakhstani 'Sante' cultivars resulted in favorable amount when sowing 2∼3 tubers with 5∼10g of tuber. Therefore this showed necessity of exportation.

It was investigated that the climatic environment (temperature, precipitation, sunshine hours) of Vietnam, Mongolia, China and Uzbekistan, which are the countries to be exported for potato cultivars of Korea during 2012 ∼ 2014. Through these data, it was established that cultivation environment of potato and settlement of storage period for potato cultivation. Cutivars for this experiment was cv. Atlantic. Sprout inhibitor(CIPC) was applyied for long-term preservation (preservation for 6 months, storage temperature 25℃.), it was treated on the basis of days (0, 10, 20, 30 days) and a kind of light wavelength to induce dormancy was applyied. It was examined the effect of yellow light treatment on physiological vitality. At 6 month after applying of CIPC and yellow light, the degree of physiological vitality was analyzed by investigating the sprouting rate and the sprout length. The effect of sprout inhibitor and Yellow light on productivity of seed tuber was carried out through pot cultivation. Finally, in order to verify whether seed potato through pot cultivation can be applied in actual open field cultivation, it was carried out the stability and productivity the seed potato cultivated in the open field.

1. Selection of the problem pests by monitoring the occurrence of pests in export target countries.
We have selected the most harmful pests for potatoes in Vietnam, Mongolia, China, Uzbekistan and Kazakhstan, which are major export destinations. China (Heilongjiang Province) suffered a great deal of damage from late blight. Mongolia and Central Asia (Uzbekistan and Kazakhstan) have the greatest damage from early blight. Especially in Central Asia, the damage of Colorado potato beetle was serious. In Viet Nam, the damage of the disease such as late blight and bacterial wilt was serious and various insect pests were generated. In order to export to Vietnam, it is necessary to develop cultivars resistant to late blight and bacterial wilt. In Central Asia, countermeasures against Colorado potato beetle are needed.

2. Development of potato pest resistance evaluation system and support for resistance evaluation
In the case of late blight, we have developed a novel resistance assay using leaves and tubers. We have developed an evaluation method based on the average reaching days that can be objective and absolute in the existing subjective and state evaluation. It is a technique to evaluate the resistance by examining the point where 100% late blight is reached by making leaf and tuber the same size. The resistance evaluation standard was also divided into 5 steps and redefined so that everyone can understand it easily. A system for evaluating field resistance in a glass house was constructed to ensure accurate resistance evaluation. We have used the developed technology to determine resistance to genetic resources and breeding lines.

Bacterial wilt disease is caused by Ralstonia solanacearum and it can cause major economic losses in particular with commercial potato production worldwide with a high impact on mass cultivated areas. To date, however, there are no solutions to prevent or cope against this disease. Alternatively, resistant cultivars of potato, which hybrid with wild potato sources seem thus to be more convenient and efficient way to control bacterial wilt disease. However, there are limited methods available to screen strong resistant cultivars because of variations of ability of the pathogenicity by each pathogen strain and their favorable conditions to induce symptoms. Thus, breeders are frequently faced up with problems that they do not know what cultivar is most resistance from lines they produced.

Therefore, in this study, we evaluated standard for disease index, which is scored at 5 degrees based on symptomatology of inoculated seedlings from sprout and tissue culture using 5 popular cultivars such as Daeji, Daeseo, Seohong, Sumi, and Haryeong. Bacterium is inoculated by pour- and clip-inoculation and optimal recording day is 9 and 12 days post inoculation (dpi) for sprout and tissue culture seedlings, respectively. Daeji and Daeseo showed more resistance and then Seohong and Sumi were moderate, and Haryeong appeared less resistance on the basis of standard scheme from both seedlings. These results suggest that our disease index may play a critical role in vitro screening for resistant lines. Additionally, we did blind test using 54 breeding lines by clip-inoculation method. Resistant scores were almost similar levels to those of previous studies, indicating that disease index is sufficient to screen massively. Further, this disease index is easy to use for researchers who are not expertise on this field. The overall screening results would help breeders to develop easily bacterial wilt disease-resistant potato cultivars.

Streptomyces spp., including scabies, turgidiscabies, and acidiscabies are major causal agents of common scab disease on potato tubers. To date, however, several strains that shown different characters compared to those strains, such as colony color and chain morphology, have been reported as minor pathogens to cause common scab disease worldwide. This means that Streptomyces spp. have continuously shown phenomena to gain virulence factors as form of pathogens. In the present study, we tested several methods to determine of which cultivars are resistant against common scab pathogens such as using potato disk assay with strains and extracts of those strain. We also tested feasibility of the inoculation of crude extracts on sprout of tissue culture of 5 popular cultivars such as Daeji, Daeseo, Seohong, Sumi, and Haryeong. However, there is no correlation among tested methods and resistance of 5 potato cultivars, indicating these methods were not appropriate to evaluate resistant cultivar against common scab disease.

Thus, we developed alternatively using synthetic thaxtomin D that useful in case of quantification of inoculum to develop disease scores. In terms of synthetic thaxtomin D, we have enough amount to test 3,000 sprouts at all and confirmed minimum concentration for this test. Nowadays, we are conducting test of which cultivars are resistant to common scab disease.

3. Evaluation of drug resistance of major insect pests of export target countries and investigation of drug use status
The survey of major pests at the potato cultivation area was conducted in Vietnam, Kazakhstan and Mongolia. The thrips and aphids were major pest in Vietnam and Colorado potato beetles was dangerous pest in Kazakhstan. Interestingly, a few pest was found in Mongolia due to the high population density of natural enemies.

In the susceptibility comparison of phosphine at different stages, it was more sensitive in the order of larvae, adults, eggs, and pupae. CT values to control all stages were resulting in 40 mg ·h/ℓ and 60 mg·h/ℓ at 20℃ and 5℃, respectively. Higher concentration of oxygen revealed the higher toxcity. The ethyl formate showed higher toxicity at adult than larvae and the appropriate CT value to control all stages was 300 mg ·h/ℓ at 20℃ and 5℃, respectively. There was no cooperative effect by oxygen.

In order to control the Colorado potato beetle in Kazakhstan, we evaluated the resistance level by egg toxicity methods. Organophosphate and chlorantraniprole revealed the high toxicity. In addition, the resistance frequency was determined using the previously identified resistance marker on AChE and VSSC, respectively. The resistance allele frequency associated with organophosphorus was high in all regions, while the pyrethroid resistance level was revealed high divergence by regions. It is necessary to decide the appropriate pesticide after assessment resistance level for the proactive management of Colorado potato beetle.

Colorado potato beetles are a very dangerous pest in Kazakhstan. Especially in the mid 1990 's, it appeared to have invaded from abroad. By the population genetic analysis using AChE2 variation, it is thought that the population of Kazakhstan came from Poland rather than Russia.

4. Development of antibody production technology for mass production of potato virus diagnosis kit
In order to develop in vitro antibody production technology, an expression and purification system for virus antibody protein production was developed. The scFv library responsive to PVY was selected and a plasmid vector for recombinant antibody expression was constructed. When protein A was used for purification, a non - specific reaction occurred when the recombinant antibody was at a high concentration, and was changed to a His tag. The purification system using His tag was constructed and protein expression of the recombinant antibody was confirmed. For the production of recombinant antibodies and the production of virus diagnostic kits, recombinant antibody plasmids labeled with enzyme proteins were developed. Since a secondary antibody with a chromogenic enzyme protein is required in the diagnostic kit such as ELISA, three recombinant phagemid vectors (HRP, GFP, ALP) capable of producing the recombinant phagemid vector have been developed. The immunostrips were prepared using the recombinant antibodies produced and the effects were tested. We have produced a prototype of ELISA and immunostrip for virus detection

(출처 : SUMMARY p.8)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 요 약 문 ... 3
• SUMMARY ... 8
• CONTENTS ... 13
• 목차 ... 14
• 제 1 장 과제의 개요 및 성과목표 ... 15
• 제1절 과제의 개요 ... 15
• 제2절 성과목표 ... 16
• 제 2 장 국내외 기술개발 현황 ... 17
• 제 3 장 연구개발수행 내용 및 결과 ... 19
• 제1절 연차별 연구개발 수행내용 ... 19
• 제2절 연구결과 ... 25
• <제1세부프로젝트> 씨감자 생산기술 및 생리적 활력 조절기술 개발 ... 25
• <제2세부프로젝트> 씨감자 대량생산 파일롯 시스템 개발 ... 100
• <제3세부프로젝트> 병해충 저항성 평가 및 관리기술 개발 ... 139
• 제 4 장. 목표달성도 및 관련분야 기여도 ... 216
• 제 5 장. 연구개발 성과 및 성과활용 계획 ... 219
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 222
• 제 7 장 참고문헌 ... 224
• 끝페이지 ... 232