담배와 벼의 발아와 생장에 대한 Proton 빔조사의 영향

배창휴(Chang-Hyu Bae),류재일(Jae-Il Ryu),사란투야 젠다람(Gendaram Sarantuya),채종서(Jong-Seo Chai),김재홍(Jae-Hong Kim),양태건(Tae-Gun Yang),이민용(Min-Yong Lee),양덕춘(Deok-Chun Yang) 한국자원식물학회 2005 한국자원식물학회지 Vol.18 No.3

Effects of proton beam irradiation on seed germination and growth pattern of tobacco (Nicotiana tabacum L. cv. BY-4; N. plumbaginifolia) and rice (Oryasativa L.) plants were estimated to develop the efficient conditions of irradiation. Seed germination rate was decreased by increasing the proton beam the current and the beam irradiation time in both tobacco and rice seeds. The beam irradiation conditions showing 50% germination were over 60 sec at 10 nA, approximately 5 sec at 100 nA and at 500 nA beam current in tobacco seeds. And the conditions of 50% germination were 60 sec at 10 nA, and 100 nA and 30 sec at 500nA in rice (cv. Dongjin 1) seeds. The growth of irradiated plants was decreased, but significant difference in morphological changes was not observed by the proton beam treatment. The proton beam is able to use as a mutagen, but some of the factors including beam size and beam detector-system must be established for efficient usage of the beam.

재배지 및 야생지에서 국내 차나무(Camellia sinesis L. O. Kuntze)의 유전적 다양성

배창휴(Chang-Hyu Bae),김길자(Kil Ja Kim),문승옥(Seung Ok Mun),류재혁(JaiHyunk Ryu) 한국차학회 2015 한국차학회지 Vol.21 No.2

In this study, ISSR markers were applied to assess the genetic diversity and structure analysis of tea (Camellia sinesis L. O. Kuntze) among the 3 cultivated populations, a wild population and single variety population (cv. Yabukita) as a control in Korea. Out of 129 loci detected overall, 120 were identified as being polymorphic with a rate of 93.02% in the 50 individuals from the populations. The polymorphic rate of the within population was over (more than) 80% in the four populations, but 16% in the single variety population. The mean of Nei's gene diversity (h) and Shannon‘s information index (S.I.) of the cultivated populations were lower than that of the wild populations. The high values for the total gene diversity, effective and Nei’s genetic diversity indicated substantial variations among the cultivated population. In addition, the insights into the relative gene diversity among and within population of Jeonggeum-ri would be useful in breeding and for the development of strategies for tea genetic population. Based on the cluster analysis of UPGMA, the genetic relationship among the all populations was coincided with the pattern of cultural environment.

Neutron 빔조사 담배 및 벼식물체의 특성

배창휴(Chang-Hyu Bae),채종서(Jong-Seo Chai),김재홍(Jae-Hong Kim),양태건(Tae-Gun Yang),류재일(Jae-Il Ryu),이효연(Hyo-Yeon Lee),양덕춘(Deok-Chun Yang) 한국자원식물학회 2005 한국자원식물학회지 Vol.18 No.3

재배종담배(Nicotiana tabacum L. cv.), 야생종담배 (N. plumbaginifolia)와 벼종자(Orya sativa L, cv.)에 neutron빔을 각각 조사(irradiation)하여 발아, 식물체의 생장과 DNA의 변이에 미치는 영향을 검토하였다. 담배종자와 벼종자에 90, 180, 270, 360, 450, 540 Gy까지 조사로 발아율은 크게 감소하지 않았고, 생장에 있어 유의적인 차이는 없었다. 그러나 담배식물체에서 총 200개체 중 71개체(약 36%)가 잎에서 형태 이상을 나타내었다. 또한 줄기색 변이체, 엽색변이체, 화형변이체를 유도하였다. 이 결과는 neutron빔이 유용한 돌연변이원으로서 가능성이 있음을 시사해 준다. Neutron빔을 조사한 후 생장한 담배식물체의 잎에 대하여 총 34개의 primer를 이용하여 RAPD 분석한 결과 20개의 primer에서 총 104개의 DNA 단편이 증폭되었고, 중성자 빔조사처리구에서만 출현하는 DNA 단편은 나타나지 않았다. Effects of neutron beam irradiation on seed germination, growth and RAPD pattern of tobacco (Nicotiana tabacum L. cv.; N. plumbaginifolia) and rice (Orya sativa L. cv.) plants were estimated. Seed germination rate was not significantly changed by the neutron beam treatment in both tobacco and rice seeds. And there was no significant differance in growth of the plants by the neutron beam treatment. Interestingly, however, some of morphological changes, including leaf shape (about 36%), stem color and leaf color were observed in neutron beam treated tobacco plants. In addition, abnormal flower in petal was observed in the neutron beam treated plant. This results indicate that neutron beam is able to use as an effective mutagen in plant mutations. Scorable products from 20 primers were obtained by RAPD analysis in the leaves of the beam irradiated tobacco plants and most of the plants showed the similar band patterns.

Particle Inflow Gun을 이용한 벼 캘러스 내의 효율적 유전자 도입

송인자,배창휴,최대옥,이효연,Song, In-Ja,Bae, Chang-Hyu,Choi, Dae-Ock,Ryo Akashi,Lee, Hyo-Yeon 한국식물생명공학회 2002 식물생명공학회지 Vol.29 No.4

본 연구실에서 개발된 particle inflow gun (PIG)은 조작이 간편하고, 사용비용도 저렴하며, 식물 세포 내의 유전자 도입효율이 높은 특징을 갖고 있다. PIG 장비를 이용하여 벼 캘러스 내로의 유전자 도입 조건을 검토하기 위해서 사용된 vector는 pIG121Hm으로서 T-DNA 내부에 intron GUS ($\beta$-glucuronidase)와 hygromycin 및 kanamycin 저항성 유전자를 포함하고 있다. 또한 벼 캘러스 내에 물리적으로 DNA를 도입할 때에 DNA 도입 효율과 관계가 높은 요인들을 GUS의 발현빈도를 통하여 조사하였다. 그 결과 gold particle에 DNA를 부착하는 과정에 사용되는 spermidine과 calcium chloride의 경우 무첨가구에 비해 16 mM의 spermidine과 1.5 M의 calcium chloride 첨가구에서 GUS 발현율이 각각 2배, 3배 증가하였다. 그리고 1회 분사되는 gold particles양이 2 mg의 경우 가장 높은 GUS 발현율을 보여주었으며, 또한 PIG장비의 분사거리와 헬륨의 압력은 벼의 배양세포의 경우 12cm의 분사거리에서 3.5 bar (50 psi)의 헬륨압력으로 분사하였을 때 GUS 발현율이 가장 높았다. 이상의 결과에서 PIG 장비를 이용한 유전자 도입은 본 연구에서 검토한 최적의 조건을 이용하였을 경우 기존에 많이 사용되고 있는 Biolistic Gun (Bio-Rad 사)과 거의 비슷한 유전자 도입효율을 보여 주었다. 특히 PIG 장비의 경우 조작이 매우 간편하고, 분사에 사용되는 일회용 부품이 필요하지 않기 때문에 대량의 반복실험을 필요로 하는 연구에서 손쉽게 사용되리라 기대된다. We have developed a homemade particle inflow gun (PIG) system which has simple operation method, low price and high gene introduction efficiency into rice callus. Rice callus were inflowed with gold particles containing DNA of a plasmid, pIG121Hm, harboring intron GUS ($\beta$-glucuronidase) gene, NPTII gene and HPT gene. For optimal GUS transient expression, the effects of parameters on DNA delivery efficiency of the PIG system was investigated by scoring transient GUS expression. The highest number of blue spots was observed at 16 mM of spemidine and 1.5 M of calcium chloride, respectively. And the amount of gold particles required for the best GUD expression was 2 mg. Optimum GUS transient expression was observed at target distance of 12 cm and helium pressure of 3.5 bar (50 psi). Gene introduction efficiency of the PIG system was observed almost similar to that of the Biolistic Gun (Bio-Rad Company). Since PIG system is simple to operate and one doesn't need disposable accessaries, the PIG system can be easily applied to various replication experiments.

Agrobacterium tumefaciens를 이용한 벼의 형질전환 효율의 검토 및 내한성 관련 GPAT (glycerol-3-phosphate acyltransferase) 유전자의 형질전환

서미숙,배창휴,최대옥,임성렬,서석철,송필순,이효연,Seo, Mi-Suk,Bae, Chang-Hyu,Choi, Dae-Ock,Rhim, Seong-Lyul,Seo, Suk-Chul,Song, Pill-Soon,Lee, Hyo-Yeon 한국식물생명공학회 2002 식물생명공학회지 Vol.29 No.2

Agrobacterium을 이용한 벼의 효율적인 형질전환을 위하여 몇 가지 형질전환 조건을 조사하였다. 그리고 조사된 최적의 형질전환 방법을 이용하여 내한성에 관련된 GPAT 유전자를 식물에 도입하였다. 형질전환 조건은 GUS 발현을 통하여 조사되었다. 본 실험에서는 벼 (Oryza sativa L. cv. Dongjin)의 성숙 종자 유래 캘러스를 3일간 전배양한 후 Agrobacterium을 접종하였다. 접종이 끝난 캘러스는 50mg/L CaCl$_2$, 30mg/L acetosyringone, 2 mg/L 2,4-D, 120 mg/L betaine이 첨가된 공동배양 배지 위에서 암조건으로 10일간 공동배양하여 높은 GUS 발현을 관찰할 수 있었다. 이와 같은 방법으로 GPAT 유전자를 식물에 도입한 결과 54%의 높은 형질전환율을 나타내었다. 형질전환 식물체를 southern 분석한 결과 wild type 식물체에서는 GPAT 유전자가 검출되지 않았으나, 형질전환 식물체에서는 GPAT 유전자가 검출되었다. 또한 GPAT 유전자로 형질전환된 5 계통의 T1 세대에서 hygromycin에 대한 저항성과 감수성의 유전비율이 3 : 1로 분리되었다. 따라서 본 실험의 결과에서 검토된 고빈도의 형질전환 시스템은 단자엽식물의 형질전환에 있어서 모델계로 이용될 수 있으리라 기대된다. This study has been focused on improving transformation efficiency of rice using Agrobacterium tumefaciens. We have demonstrated the effect of this system when the GPAT gene related to the cold-resistance was transferred by Agrobacterium tumefaciens in rice. Transformation conditions were modified using intron $\beta$-glucuronidase (GUS) expression as a reporter gene in the rice. In this study, mature seed-derived calli of rice (Oruza sativa L. cv. Dongjin) were pre-cultured for 3 days and then infected with Agrobacterium. When this infected calli were cultured in the dark for 10 days on co-cu]lure medium containing 50 mg/L of CaCl$_2$, 30 mg/L of acetosyringone, 2 mg/L of 2,4-D, 120 mg/L of betaine, high GUS expression was observed. In the present transformation system, the efficiency of transformation of GPAT gene was about 54%. Stable integration of GPAT gene into chromosomal DNA was proven by southern blot analysis of genomic DNA isolated from T$_{0}$ progenies. The progenies (T1 generation) derived from primary transformant of 5 lines were segregated with a 3 (resistant) : 1 (sensitive ratio) in medium containing hygromycin. This high frequency transformation system can be used as a useful tool in transformation of another monocotyledon.n.

차나무의 맹아와 신초생장에 미치는 온도의 영향

이선하(Seon-ha Lee),배창휴(Chang-Hyu Bae) 한국차학회 2005 한국차학회지 Vol.11 No.1

There are light intensity, temperature, CO2, humidity, and fertilization in the environmental conditions of greenhouse cultivation of tea. The influence of temperature on bud break and shooting of tea was studied in this study. The bud breaking of tea was increased as the temperature went up. The buds of all cuttings were broken in 15 days when the cuttings had been placed in a growth chamber at 30℃. It took 27 days for the cuttings kept at 20℃. Under the natural condition, the buds of tea plants broke at April 10th. When the cuttings were moved from the field into a growth chamber at 25℃ starting from December 25th at 10 days' interval, the time required for bud breaking could be decreased by 5 days for every 20 days kept at the growth chamber. The shooting responses of the 5 varieties tested to temperature were similar to each other. The optimum temperature for the shooting of the tea was found to be 25℃. The photosynthesis rate increased significantly as the temperature went up to 25℃, then decreased at temperatures higher than 30℃.

무인헬기용 Macro granule 제초제의 벼 작형별 살초효과 및 주변작물에 미치는 영향

윤철수(Cheol-Su Yoon),배창휴(Chang-Hyu Bae),이성춘(Sheong-Chun Lee),김경현(Kyung-Hyun Kim),이계환(Kye-Hwan Lee),조태경(Tae-Kyoung Cho),황인천(In-Cheon Hwang) 韓國雜草學會 2011 Weed&Turfgrass Science Vol.31 No.4

무인헬기를 이용한 제초제처리는 제초제의 특성이 항공방제에 적합해야하는 등 선결 요인이 무척이나 많다. 논에 발생하는 잡초를 방제할 목적으로 환경친화적·생력형 macro granule(GG) 제초제를 개발하였다. 무인헬기를 이용한 벼 작형별 잡초방제효과 및 벼에 대한 생육특성을 조사하고, 주변작물에 미치는 영향을 평가하였던 결과를 요약하면 다음과 같다. GG의 무인헬기살포효과는 담수직파시험에서 93.6∼96.6%의 방제효과를 보였는데, 손제초구와 큰 차이를 나타내지 않았다. 수량은 손제초구와 약제처리구가 각각 4,700kg ha-1, 4,450∼4,670kg ha-1로 비슷하였다. 기계이앙시험에서는 시험약제 모두 92.7∼97.3%의 방제효과를 보였다. 무인헬기살포와 손제초구의 수량은 각각 5,270∼5,520kg ha-1, 5,730kg ha-1로 큰 차이는 없었다. 또한 GG살포가 주변작물에 미치는 영향을 보면 액상제형과 GG가 직접살포되었을 때 배추, 상추, 오이, 고추, 콩, 들깨에 생육억제, 황변, 낙엽 등의 증상이 나타났으며, 일부 개체는 고사하였는데 이것은 무인 헬기를 이용한 약제 살포의 문제점을 시사한 것으로 무인헬기 살포시 주변작물에 약액이 비산되지 않도록 주의가 필요 할 것으로 사료된다. 향후 사용 약제의 선택, 시기, 주변 작물의 종류 등을 감안 한 주도면밀한 연구가 반드시 필요하다고 생각된다. 본 연구의 결과 무인헬기를 이용한 제초제 살포는 우리나라의 농업이 처해진 여건상 새로운 대안이 될 수 있다고 보여지며 이에 대한 체계적인 연구가 더욱 심도 있게 수행되어야 한다고 생각된다. This experiment was carried out to confirm characteristics of macro granule (GG) for herbicidal efficacy by using remote-controlled aerial application (RCAA) to control annual and perennial weeds in rice paddy field, and phytotoxicity to rice and non target plants of formulation types. Herbicidal efficacy of GG applied by using RCAA was 93.6∼96.6% in flooded direct sowing cultivation. There was no difference in phytotoxicity between the hand and the power applicator methods in terms of height, number of tiller and yield component. Herbicidal efficacy of GG applied by using RCAA was 92.7∼97.3% in machine transplanting rice field. There was no difference in phytotoxicity between the hand and RCAA methods in terms of height, number of tiller and yield component. When suspension concentrate (SC) and GG were applied directly on Chinese cabbage, lettuce, cucumber, pepper, soybean and wild sesame, several symptoms of injury such as, the inhibition of growth, yellowish, leaf fall and withering was observed.

환경요소가 차나무 광합성에 미치는 영향

이선하(Seonha Lee),배창휴(Chang Hyu Bae),임준택(June Taeg Lim) 한국차학회 2003 한국차학회지 Vol.9 No.2

The effects of irradiance, temperature, humidity, and CO2 concentration on the rate of photosynthesis of one year tea plants were studied using Gas Exchange Measuring Station. The rate of photosynthesis was estimated by the CO₂ concentration in a sealed chamber containing a tea plant. The rate of photosynthesis of tea plants being cultivated in a greenhouse was also measured under full irradiating and 40% shading conditions using a portable photosynthesis tester. The light compensation point was 20-40 PPF(photosynthetic Photon Flux) and the light saturation point was higher than 900 PPF. The rate of photosynthesis of tea plants increased as temperature went up to 25˚C, and decreased at temperatures above 30˚C. The increase in decreased rate of photosynthesis. The carbon dioxide compensation point was 200ppm and the CO₂ saturation point was 1300ppm. Meiryoku showed the highest rate of photosynthesis among the 6 tea cultivars cultivated in a greenhouse. The rate of photosynthesis under 40% shading condition was 20% lower than the rate under full irradiating condition.

무인헬기용 Macrogranule(GG) 제초제의 안정성 및 살포방법 연구

윤철수(Cheol-Su Yoon),배창휴(Chang-Hyu Bae),이성춘(Sheong-Chun Lee),김경현(Kyung-Hyun Kim),이계환(Kye-Hwan Lee),조태경(Tae-Kyoung Cho),황인천(In-Cheon Hwang) 韓國雜草學會 2011 Weed&Turfgrass Science Vol.31 No.3

무인헬기 살포로 논에 발생하는 잡초를 방제할 목적으로 macrogranule(GG) 제초제를 개발하였다. 약제제조와 제조약제의 안정성, 살포방법에 따른 적용방법을 조사하였고, 제형별 살포소요시간 경제성을 분석한 결과를 요약하면 다음과 같다. 제조 GG의 입경(粒經)은 2.5~3mm 범위내에 85%이상을 차지하였고, 가비중은 0.2~0.4g mL-1로 수면에서 부유성을 갖고 확산되는 입제고형물이었다. GG의 저장안전성은 54±℃ 항온기온에서 2, 4, 6, 8주간 저장 후 halosulfuron-methyl을 HPLC로, mefenacet는 GC로 분석한 결과 유효성분 및 외형 변화가 없이 안정되어 있었다. GG의 수중붕괴성은 온도가 상승할수록 증가하였다. 또 담수 중 pH별 수중붕괴성은 담수 pH의 영향은 없었다. GG의 살포거리는 손살포, 도구살포 및 기계살포 등에서 각각 4~5m, 7m, 10~12m로 살포방법에 따라 달랐다. GG의 약효 및 작물안전성은 손살포, 도구살포, 기계살포 및 무인헬기살포에서 모두 우수하였다. 제형간 4,000㎡ 기준 손살포 소요시간은 GG, GR, SC, TB가 각각 38분 4초, 42분 20초, 38분 10초, 21분 4초였으나, 무인헬기 살포시 1분 32초를 나타냈다. 제형의 형태 및 특성상 무인헬기살포가 가능한 제형은 SC, GG였다. This experiment carried out to confirm characteristics of macrogranule (GG) for herbicidal efficacy by using remote-controlled aerial application (RCAA) to control annual and perennial weeds in rice paddy field, application methods, and application times of formulation types. Particle of 500 g GG having average diameter of 2.5∼3 mm was over 85 percent, the bulk density of the particle was 0.2∼0.4 g mL-1 and the particle was water floating granule. Active ingredients and external form of halosulfuronmethyl+mefenacet GG remained stable under condition of storage stability test. The disintegration time of the GG was faster as the water temperature was increased. But disintegration time was not affected by pH on the water conditions. By using hand, spoon and power applicator, drift distances of GG were 4∼5 m, 7 m and 10∼12 m, respectively. GG showed good herbicidal efficacy and plant safety in all of the application methods such as, using hand, spoon, power applicator and RCAA. Application times of GG, GR, SC and TB by hand in 4,000 m² were 38 min. 4 sec, 42 min. 20 sec, 38 min. 10 sec and 21 min. 4 sec, respectively, but application time of GG by using RCAA was 1 min 32 sec. According to appearance and characteristics of formulation types, suspension concentrate (SC) and GG were possible formulation types for RCAA.

ISSR 분석에 의한 구례지역 차나무 집단의 유전다양성과 구조

류재혁(JaiHyunk Ryu),문승옥(Seung Ok Mun),배창휴(Chang-Hyu Bae) 한국차학회 2011 한국차학회지 Vol.17 No.1

In this study, ISSR markers were applied to assess genetic diversity and structure analysis of four tea (Camellia sinensis) populations at Gurye region in Korea. Out of 141 loci detected overall, 135 were identified to be polymorphic with a rate of 95.7% in the 60 individuals. The mean of Nei’s gene diversity (h) and Shannon’s information index (S.I) were 0.230 and 0.350, respectively. AMOVA indicated that 20.4% of the variation was due to the between population and the remaining 79.6% to the within population. The values of total genetic diversity (H_t), average gene diversity within population (H_s), and proportion of total genetic diversity partitioned among populations (G_ST) by Nei’s (1978) method were 0.284, 0.229, and 0.194, respectively. Interpopulation gene flow (Nm) of 2.06 (Nm〉1.0) indicates that genetic exchange occurred among the populations. The average genetic distances among the populations were 0.113 Hwaeomsa, 0.089 Piagol, 0.083 Jirisanplaza and 0.074 Choneunsa, respectively. Based on the cluster analysis of UPGMA, genetic relationship among the four populations was not coincide with the pattern of geographic distribution.