초록 ▼

Ⅳ. 연구개발결과
1. 명품마늘재배용 친환경 미생물 제제 개발
가. 동절기 충청북도 농업기술원 실험 포장지역의 토양을 채집하여 분리한 균주 중 저온적응성을 확인하여 총 66 종의 균주를 1차 선발하고 형태, 생리학적 특성을 조사.
나. 1차 선발 내한성 미생물의 16S rRNA 염기서열 분석을 통하여 총 40개의 균주를 2차 선발한 결과, 22개의 균주는 Pseudomonas 속과 염기서열이 유사하였으며, 7개의 균주는 Flavobacterium 속, 6개의 균주는 Arthrobacter 속, 3개의 균주는 Mass

Ⅳ. 연구개발결과
1. 명품마늘재배용 친환경 미생물 제제 개발
가. 동절기 충청북도 농업기술원 실험 포장지역의 토양을 채집하여 분리한 균주 중 저온적응성을 확인하여 총 66 종의 균주를 1차 선발하고 형태, 생리학적 특성을 조사.
나. 1차 선발 내한성 미생물의 16S rRNA 염기서열 분석을 통하여 총 40개의 균주를 2차 선발한 결과, 22개의 균주는 Pseudomonas 속과 염기서열이 유사하였으며, 7개의 균주는 Flavobacterium 속, 6개의 균주는 Arthrobacter 속, 3개의 균주는 Massilia 속, Flavimonas 속 1개와 Pedobacter 속 1개의 균주가 유사한 염기서열을 보이는 것을 확인.
다. 2차 선발 내한성 미생물의 생장온도 범위, 상대 생장계수 (RGRC), 지방산 메틸에스터 분석 (FAME)을 실시한 결과, 총 40개의 균주 중에서 10 개의 균주 (Pseudomonas 8, Flavobacterium 2)가 내한성 미생물 고유의 특징 (최대 생장온도 35℃, 상대 생장계수 0.5이상, 3개 이상의 지방산 함유)을 모두 충족하는 것을 확인.
라. 2차 선발 내한성 미생물의 식물생장촉진 특성을 분석한 결과, 총 40개의 균주 중 24개 균주에서 IAA 생성능을 보였으며, 인산가용화능은 19개 균주, ACCD 생성능은 9개 균주, Siderophore 생성능은 13개 균주에서 확인.
마. 저온 (15℃)에서 내한성 미생물의 접종에 따른 작물 종자의 발아율을 분석한 결과, 총 40개의 균주중에서 30개의 균주가 종자의 발아율을 증가시켰으며, 그 중에서 12개의 균주 (OR309, OS114, OS115, OS124, OB134, OB138, OB342, OB145, OS253, OB155, OS261, OS263)를 접종하였을 때 종자의 발아율이 유의성 있는 증가를 보이는 것을 확인.
바. 저온 (15℃)에서 내한성 미생물 (OB115, OS253, OB155, OS261)의 접종에 따른 유묘의 생장은 확인한 결과, 균주를 접종하지 않은 처리구에 비해 균주를 접종하였을 때 유묘의 생장이 증가하였다. 뿌리 길이와 지상부 길이는 OS155 균주를 접종하였을 때 유의성 있는 증가를 보였으며, 균주를 접종하지 않은 처리구에 비해 각각 10.3%, 16.3% 증가하였음.
사. 저온 (15℃)에서 내한성 미생물 (OB115, OS253, OB155, OS261)의 접종에 따른 식물조직 내 프롤린 축적량을 확인한 결과, 균주를 접종한 처리구에서 균주를 접종하지 않은 처리구에 비해 프롤린 축적량이 증가하였으며, OS261 균주의 경우 프롤린 축적량이 91.2±11.2 nmol g-1 FW으로 유의성 있는 증가를 확인.
아. 응집력 향상을 위해 선택한 HCN-AMS 배지에서 배양한 모든 실험 균주가 높은 성장률 및 응집 형성에 의한 높은 건중량을 보임.
자. 실험 균주의 EPS 분석 결과, 모든 균주들이 형광을 띄는 양성의 반응을 보였으며, HCN-AMS 배지에서 배양한 균주에서 가장 높은 EPS 생산량을 확인.
차. 실험 균주의 PHB 분석 결과, HCN-AMS 배지에서 배양한 균주에서 가장 높은 PHB 축적량을 확인.
2. 명품마늘 생산을 위한 친환경재배기술 개발
가. 마늘 친환경 재배에서 주아를 이용한 종구생산체계가 우량종구를 생산할 수 있다. 이때 주아를 이용한 종구생산 시 가장 중요한 해충이 파좀나방이며, 이 해충을 방제하기 위해 페로몬트랩을 이용한 성충조사와 방제약제 선발 시험 수행.
나. 단양의 4개 지역에서 파좀나방을 조사한 결과 5월 상순부터 발생되며, 5월 하순~6월 상순에 최고 밀도를 보이고, 그 후 감소됨. 페로몬 트랩은 7일 간격으로 깨끗한 트랩으로 교체하여 성충을 조사
다. 파좀나방에 효과가 있는 생물농약과 유기농자재를 5월 31일, 6월 7일에 약제살포
(1) 생물농약 및 친환경제재는 파좀나방 알과 번데기에는 효과가 떨어지기 때문에 파좀나방 유충 시기가 형성되는 5월 하순부터 방제 시작
(2) 마늘수확이 6월 17～20일 사이이고, 주아 수확은 3～5일정도 전에 하는 것이 일반적이기 때문에, 마늘에서 파좀나방 마지막 약제처리는 6월 10일 이전임
라. 친환경 약제처리 후 파좀나방 방제효과 조사는 성충 조사가 어려워, 유충과 번데기의 밀도로 6월 27일과, 7월 5일 조사함
마. 방제약제는 비티아이자와이와 비티쿠르스타키가 우수한 약효를 보이나, 이 약제는 생물농약으로 등록되어 있고, 유기농자재로 등록되어 있지 않아 무농약 재배에서만 사용이 가능함. 따라서 유기농재배로 석회유황 합제, 난황류3%, 목초액(300배)을 사용하고 무농약재배로 비티아이자와이(상표명:젠타리, 미성살충탄), 비티쿠르스타키(상표명: 비티, 바이오비트, 그물망, 비결, 바이충, 스콜피온케이) 사용함
3. 명품마늘 생산지의 최적 토양관리 체계 확립
가. 용적밀도가 낮아질수록 토양 열 전달율은 낮아져 보온성이 높아지고 마늘의 종구 크기는 증가하였음.
나. 토양 용적밀도를 1.0 g cm-3 과 0.8 g cm-3 로 조절할 경우, 동절기 보온성은 2~15% 정도 상승하여 마늘 종구의 발아와 생육을 촉진하였으나, 토양 수분 보유능이 감소하여 구비대기 한발피해에 민감할 가능성이 있음.
다. 토양 용적밀도 조절용으로 코코피트, 우드칩과 같은 고 C/N 유기자재를 사용할 경우, 질소기아와 같은 영양부족 상태는 유발되지 않았으나, 퇴비를 사용할 경우, 염류집적, 영양불균형, 가스장애 등에 의해 생육저해가 발생하였음.
라. 동절기 보온성 효과를 위해 토양 표면에 유기물을 멀칭 처리 (코코피트, 우드칩)하였을 경우, 기대보다 효과가 미미하였고 유기물을 토양에 직접 혼입하여 용적밀도를 조절하였을 경우, 동절기 보온성에 긍정적으로 기여하였음.

Abstract ▼

○ Title : Development of eco-friendly micro organism for high quality garlic production
Soils that belong to the temperate regions are subjected to seasonal drop in temperature for a considerable portion of a year, during which, any biological activity is low or even negligible due to intermitten

○ Title : Development of eco-friendly micro organism for high quality garlic production
Soils that belong to the temperate regions are subjected to seasonal drop in temperature for a considerable portion of a year, during which, any biological activity is low or even negligible due to intermittent dropping of soil temperatures to suboptimal ranges. This has led to short cropping periods for agriculture in the temperate regions. Cold stress causes a major metabolic response in plants, affecting their growing periods and consequently resulting in loss of agricultural production. In the present was aimed at chilling stress alleviation in plants through use of cold-adapted plant growth promoting bacteria. Soil samples were collected from fields of Chungcheongbuk-do Agricultural Research and Extension Services, Ochang-eup, South Korea during late winter season. Soils were non saline in nature (EC 0.04∼0.20 dS m^-1) and neutral in pH values (7.0∼7.5). Analyzing other soil parameters showed that organic matter content in the samples ranged from 0.8∼1.5%, total nitrogen was 0.1%, available phosphate was estimated to be at a range of 638.6∼863.3 mg kg^-1, Na 0.2 cmolc kg^-1, K 4.3∼8.3 cmolc kg^-1, Ca 0.4∼1.6 cmolc kg^-1, Mg 0.2∼0.3 cmolc kg^-1 and Fe was found to be at a range of 7.7∼8.5 cmolc kg^-1. On enrichment and selective incubation at 5℃ a total of 66 isolates were obtained on incubation of the plates at temperature of 5℃. Successive sub-culturing and growth of the isolates led to reduction of the isolate number to 63 isolates. Twenty four strains among the isolates showed production of auxin IAA in the media, 19 bacteria were found to solubilize insoluble phosphate, siderophore production was observed in 13 isolates, 9 strains exhibited ACC deaminase enzyme activity and salicylic acid was found to be produced in-vitro by 15 isolates. Based on these plant growth promotional characteristics, 40 strains were selected and genomic DNA was isolated and 16S rRNA gene amplified to deduce their phylogeny. Among the 40 isolates, genus Pseudomonas accounted for 22 strains, genus F lavobacterium was represented by 7 isolates, 6 isolates belonged to genus Arthrobacter followed by 3 strains of M assilia, 1 strain of F lavimonas and 1 strain representing genus Pedobacter.
Psychrotrophic nature of the isolates were studied in terms of the temperature range of growth (5℃ to 40℃), relative growth rate co-efficient (RGRC) values and fatty acid methyl ester analysis (FAME) to identify characteristic fatty acids that help bacteria to adapt to lower temperature conditions. Out of the forty isolates studied, 10 strains (Pseudomonas 8, F lavobacterium 2) exhibited high adaptive characteristics of psychrotolerance by having maximum growth temperatures below 35℃, RGRC values ≤ 0.5 and exhibiting at least 3 of the characteristic fatty acids for membrane adaptation. Twenty four isolates belonging to P seudomonas (14), Arthrobacter (2), F lavobacterium (4), F lavimonas (1), M assilia (3) satisfied at least 2 of the three experimental studies to analyze psychrotrophic nature. Six isolates including strains belong to Arthrobacter (4), F lavobacterium (1), and P edobacter (1) exhibited one of the studied properties for psychrotolerance. Plant studies showed that stains Arthrobacter oryzae OS115, P seudomonas frederiksbergensis OS253, P. vancouverensis OB155, and P . frederiksbergensis OS261 improved germination under a chilling stress of 15℃. When the strains were inoculated for in seedlings, they were found to promote plant growth under chilling temperature conditions by improving root and shoot lengths, dry weight and proline accumulation. Strains inoculated with P. frederiksbergensis OS253, significantly improved shoot length and plants inoculated with P. vancouverensis OB155, and P. frederiksbergensis OS261 showed high proline accumulation among the studied strains. These cold-adapted plant growth promoting bacteria can by successfully employed to alleviate chilling stress occurring in plants under sustainable agricultural practices.
Exopolysaccharide (EPS) production, PHB accumulation and stress endurance efficiency of M ethylobacterium strains CBMB20, CBMB27, CBMB35 and CBMB110 were studied upon. Growth of the strains were compared on ammonium mineral salt (AMS) medium and high carbon: nitrogen AMS (HCN-AMS) medium. Highest difference in growth on use of the two media was observed in strain CBMB110 with a 4.2% increase in HCN-AMS compared to growth in AMS medium. This trend was also followed upon by CBMB35 CBMB20, and CBMB27 at 1.9, 0.9 and 0.4% growth increase on using HCN-AMS medium. Floc yield increased by 80.0, 52.1, 93.0, and 65.2% in CBMB20, CBMB27, CBMB35, and CBMB110 respectively when HCN-AMS medium was used. Highest floc yield was 8.5 mg mL^-1 by strain CBMB35. On qualitative detection of EPS through Calcofluor white, all the M ethylobacterium strains found to be positive for EPS production. Quantification of EPS showed that M ethylobacterium strains CBMB20, CBMB27, CBMB35 and CBMB110 recorded EPS content of 0.34, 0.23, 0.25, and 0.24 mg mL^-1 in AMS medium and 0.35, 0.41, 0.35, and 0.24 mg mL^-1 in HCN-AMS medium respectively. Highest EPS production was 0.41 mg mL^-1 by strain CBMB27. Sudan Black staining to identify PHB as refractile cytoplasmic inclusions (RCI) in the M ethylobacterium strains showed that all the tested strains accumulated PHB granules when grown in HCN-AMS media. M ethylobacterium strains CBMB20, CBMB27, CBMB35 and CBMB110 recorded PHB content of 0.56, 1.15, 0.83, and 0.78 mg mL^-1 in AMS medium and 0.56, 1.20, 1.13, and 0.87 mg mL^-1 in HCN-AMS medium respectively. Highest PHB production was 1.20 mg mL^-1 by strain CBMB27. Studying the osmotic stress tolerance in the isolates, highest decrease in population was observed at 250 mM NaCl, when CBMB20, CBMB27, CBMB35 and CBMB110 populations decreased by 5.3, 3.2, 1.6, and 2.6% respectively in HCN-AMS medium and 6.0, 3.5, 7.3, and 7.9% in AMS medium. In case of acid stress, growth was arrested at pH 4.0 regardless of the media used. Acid stress at pH 5 caused 3.6, 5.9, 4.2, and 7.7% decrease in population of CBMB20, CBMB27, CBMB35 and CBMB110 in AMS medium and 0.9, 2.4, 1.3, and 4.1% in HCN-AMS medium respectively. These properties of EPS production and PHB accumulation help the bacteria in surviving under stress conditions and thereby survive and stimulate plant growth under abiotic stress conditions.
○ Title : Development of eco-friendly cultivation systems for high quality garlic production
Monitoring for Acrolepiopsis sapporensis Matsumura using sex pheromone (Z11-16Ac 0.73mg; Z11-16Al 0.18mg; Z11-16OH 0.09mg) trap from early-May to early July in 2013 were observed the occurrence of A. sapporensis adult from early May. Optimal occurrence of A. sapporensis was peaked from late May to early June. Control was the effective when spraying twice during late May to early June, the season occurring the lavae. Because biotic pesticides and organic agricultural materials were decrease the activity to the eggs and pupal stage, control of A. sapporensis was good effect at lavae stage. Among pesticides, B.t.subsp.aizawai, B.t.var.kurstaki were shown control value over 90% that excellent to control of this pest.
○ Title : Development of Optimal Soil management systems for high quality garlic production
The research of garlic grown is focus on the quality and quantity. but eco-friendly cultivation technology necessity is increased because of consumer demanding safe agricultural products. Eco-friendly agriculture is balanced both agriculture and environment in the form of continuous agricultural. also it is minimize to the undesirable effects. However, the most of present researches of garlic growth is not enough to ensure eco-friendly cultivation technology development. Therefore, the establishment of eco-friendly cultivation system is required.
The study was conducted at Danyang garlic Laboratory field for investigate effects of soil bulk density on winter season heat insulation for field growing of garlic. Soil temperature and water contents were analyzed with data logger and switching system. Soil temperature was measured at depths of 2, 5 cm with T-type thermocouple, Soil water content was measured at depths of 0-10 cm with EC-5 moisture sensor. Real-time monitoring was done throughout the growing season. Garlic growing was measured plant height, leaf width, leaf number, leaf carbide repeatedly at 10 during the first & second half of the year. Soil bulk density was investigated with Core can (diameter : 5 cm, height : 5cm).
In the 1^st year was examined. When soil bulk density increased, garlic size tended to be smaller. Because heavy soil bulk density is a factor in resistance to garlic growth of volume and root. Therefore, low soil bulk density can increase to garlic size cleary. Also, if soil bulk density is decrease, soil heat thermal efficiency is lower. so winter season heat insulation is increased. It means that heavy soil bulk density is lower buffer capacity. Therefore, It is regarded that low soil bulk density is decreased effects of external temperature. After, in this study, diverse organic matters were mixed soil for decreasing soil bulk density. Soil bulk density was adjusted 1.0 g cm^-3, 0.8 g cm^-3.
In the 2nd year was examined. When soil bulk density is adjusted 1.0 , 0.8 g cm-3, winter season heat insulation increased 2 ~ 5 % on 1.0 g cm^-3, 3 ~ 15 % on 0.8 g cm^-3. also, it was promote garlic size. However, Soil water retention capacity was decreased. Therefore, low soil bulk density was sensitive to drought damage. Also, Using high C/N Coco peat and Wood chip for decreasing soil bulk density is not cause undernourish likely nitrogen starvation. but, Using compost is occurred growth inhibiting likely Salination, nutritional unbalance, gas injury. Therefore, organic matter needed to be control.
In the 3rd year examined effect organic matter mulching and soil bulk density adjustment using organic matter mixing on winter season heat insulation and growth of garlic. For winter season soil heat insulation, experimental field has organic matter (cocopeat, wood chip) mulched. effect of soil heat insulation between vinyl and organic matter mulching was insignificant difference but, organic matter mulching treatment tends to increase soil heat insulation than vinyl mulching treatment. effect of soil heat insulation was insignificant between cocopeat and woodchip mulching amendment. Therefore To increase the volume of soil pore had positive effected on soil heat insulation than characteristics of organic matters. In growing season, there was no nitrogen starvation and nutrient deficiency and there are insignificant soil chemical properites by organic matter amendment. Therefore, To keep soil heat in winter season, adjusting soil bulkdensity using organic matter is considered to be necessary.

목차 Contents

• 표 지 ... 1
• 제 출 문 ... 2
• 요 약 문 ... 3
• S U M M A R Y ... 9
• 목 차 ... 14
• 제 1 장 서 론 ... 15
• 제1절. 연구 개발 대상 기술의 경제적·산업적 중요성 및 연구개발의 필요성 ... 15
• 제2절. 연구개발의 목표 ... 16
• 제 2 장 국내외 기술개발 현황 ... 17
• 제1절. 연구 개발 대상 기술의 국내 연구 현황 ... 17
• 제2절. 연구 개발 대상 기술의 국외 연구 현황 ... 17
• 제3절. 국내외 연구현황 비교 및 필요 연구 분야 ... 18
• 제 3 장 연구개발수행 내용 및 결과 ... 19
• 제1절 명품마늘 재배용 친환경 미생물 제제 개발 ... 19
• 제2절. 명품마늘 생산을 위한 친환경 재배기술 개발 ... 48
• 제 3 장 연구개발수행 내용 및 결과 ... 52
• 제1절. 명품마늘 생산지의 최적 토양 관리 체계 확립 ... 52
• 제 4 장 연구개발목표 달성도 및 대외기여도 ... 77
• 제1절. 목표대비 달성도 ... 77
• 제2절. 정량적 성과 ... 78
• 제 5 장 연구개발결과의 활용계획 ... 80
• 제1절. 연구개발결과의 활용방안 ... 80
• 제2절. 기대성과 ... 80
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 82
• 제1절. 독일유기농업재단 ... 82
• 제2절. 스위스 유기농업연구소 ... 82
• 제 7 장 기타 중요 변동사항 ... 83
• 제1절. 세부 연구 책임자 변경 ... 83
• 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 84
• 제 9 장 참고문헌 ... 85
• 끝페이지 ... 92