Superoxide Dismutase와 Ascorbate Peroxidase가 엽록체내로 동시에 과대발현된 형질전환 담배의 제초제들에 대한 항산화 반응

김진석,이병회,권석윤,김윤희,김소희,조광연,Kim Jin-Seog,Lee Byung-Hoi,Kwon Suk-Yoon,Kim Yun-Hee,Kim So-Hee,Cho Kwang-Yun 한국식물생명공학회 2005 식물생명공학회지 Vol.32 No.2

Superoxide dismutase와 ascorbate peroxidase가 동시에 과대발현된 담배(CA)를 가지고 여러 제초제들에 대한 항산화 반응을 조사하였다. 온실조건 실험에서, CA 담배는 PSI 저해제로 알려진 paraquat처리에 대해서 내성이 인정되었고 그 정도는 40% 내외였다. PS II 저해제 (bromoxynil, diuron, bromacil), 엽록소 생합성 저해제 (oxyfluorfen), 카로티노이드 생합성 저해제 (fluridone)와 EPSP synthase 저해제 (glyphosate) 처리에서는 CA와 wild type간의 반응차이가 관찰되지 않았다. Paraquat와 diuron을 이용한 약광 조건의 실험에서도 온실조건의 실험결과와 유사한 정도로 paraquat처리에 대해서만 내성을 나타내었다. 온실조건에서의 엽위별 반응의 경우, 6 - 9 엽기 식물체에 paraquat를 처리하였을 때, 약제처리 당시 위로부터 3 - 4번째 전개되고 있었던 잎이 상대적으로 내성 정도가 높게 나타났다. 한편 paraquat 처리 시에 여러 농도의 ascorbic acid를 혼합할 경우, CA와 wild type 모두에서 비슷한 정도로 paraquat 활성을 경감시켰다. 결론적으로, CuZnSOD/APX의 과대발현은 photosystem I 에서 발생되는 산화스트레스에 대해서만 주로 작용하며, 다른 제초제들에 의해 발생되는 산화적 스트레스에 대해서는 소거 능력이 부족한 것으로 판단되었다. Antioxidative responses of transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts was investigated with several herbicides. In greenhouse test, tolerance of SOD/APX-overexpressed tobacco (CA) to photosystem (PS) I inhibitor paraquat was increased by about 40%. However, any response differences between CA and wild type (WT) tobacco was not observed in a treatment with PS II inhibitors (bromoxynil, diuron and bromacil), chlorophyll biosynthesis inhibitor(oxyfluorfen), carotenoid biosynthesis inhibitor (fluridone) and 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase inhibitor (glyphosate). This tendency was also similar in the growth chamber test of low light intensity, using paraquat and diuron. That is, increased antioxidant activity of CA was shown only in paraquat treatment. When paraquat was foliar-treated to 6 to 9-leaf stage plant, the third to fourth placed leaf from shoot tip showed relatively higher antioxidant activity. Ascorbate supplemented to paraquat solution alleviated the phytotoxicity with a similar range in both CA and WT. In conclusion, CA specifically responded to oxidative stress induced by paraquat among tested herbicides in a whole plant assay.

백화옥수수 잎절편을 이용한 안토시아닌 생합성 조절제 탐색용 신규 검정법 확립

김진석,이병회,이정애,오광훈,조광연,Kim, Jin-Seog,Lee, Byung-Hoi,Lee, Jung-Ae,Oh, Kwang-Hoon,Cho, Kwang-Yun 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.2

This study was carried out to establish a novel bioassay system for screening of compounds affecting aromatic amino acid or anthocyanin biosynthesis through investigating a degree of sucrose-induced anthocyanin formation such as size of plant material, buffer conditions, light intensity and irradiated duration, incubation temp., etc were determined and standard procedure (suitable experimental condition) was set up as follows. The second leaf blade of white corn seeding induced by fluridone treatment were segmented into a size of 5${\times}$5 min. The segments were floated on the solution of 1% sucrose in 1.0mM MES buffer (pH6.0∼6.5) and incubated at 26$^{\circ}C$ for 2days under the continuous light condition(70∼100$\mu$mol m$^{-2}$ s$^{-1}$ ). Anthocyanin in the purpled tissues was extracted with methanol containing 1% HCl and the optical density of the clear supematants was determined at 528mm. Influences of some chemicals were tested using this system. Glyphosate, 5-enolpyruvylshikimate 3-phosphate synthase inhibitor, showed most sensitive response with I$_{50}$ value at 3.3$\mu$M. Dicyclohexylcarbodiimide(DCCD) and parachloromercuribenzenesulfonic acid(PCMBS) had a relatively strong ingibition with I50 value at 7.1$\mu$M and 10.2$\mu$M, respectively. These results show that sucrose-induced anthocyanin formation in white com leaf segment provide a very simple and rapid system for searching new compounds affecting aromatic amino acid or anteocyanin biosynthesis by screening at less than 10$\mu$M.

구멍갈파래(Ulva australis) 해조류 사멸에 미치는 여러 물리화학적 요인들의 영향

김진석,곽화숙,김보관,Kim, Jin-Seog,Kwak, Hwa Sook,Kim, Bo Gwan 한국잡초학회 한국잔디학회 2017 Weed & Turfgrass Science Vol.6 No.3

전세계적으로 해양녹조 대발생(green tide)은 해양생태계에 큰 위협이 되고 있으며 주로 갈파래과의 Ulva 속 식물이 대부분을 차지하고 있다. 본 연구에서는 구멍갈파래(Ulva australis, ULAUS) 사멸에 미치는 여러 물리화학적 요인들의 영향 정도를 조사하여 향후 갈파래 대발생 경감기술 확립을 위한 기초자료를 얻고자 제반 실험을 수행하였다. 순수에 8시간 침지는 아무런 영향을 받지 않았으나 염도 1.0-1.5% 수준에서 7일간의 저염화 처리는 ULAUS 포자 유출을 약 70% 경감시키는 효과가 있었다. 건조처리에 의해 초기 생체중의 40-50% 이상이 감소되면 심각한 생육 장해가 일어났다. 온도처리의 경우, ULAUS는 $30^{\circ}C$ 이상부터 생육에 심각한 영향을 받았으며 $35^{\circ}C$처리에서는 배양개시 1일만에 95% 이상 고사되었고, $40^{\circ}C$, $45^{\circ}C$, $50^{\circ}C$ 처리에서 엽절편이 90-95% 고사되는데 소요되는 처리시간은 각각 10분, 30초, 1초 이었다. pH 6 이하의 조건은 ULAUS 생육을 현저히 억제하였으며 pH 4에서는 신속히 사멸되었다. 한편 ULAUS 생육을 저해하는 10여종의 화학물질을 선발하였고 이들 중 구멍갈파래를 보다 신속히 사멸시킬 수 있는 화합물은 과산화수소를 비롯한 산화제들이었다. 이들의 결과를 종합해 볼 때, 갈파래 대발생제어에 저염화, 건조, 온도 및 화합물 처리 등이 실용화 기술개발에 동원될 수 있을 것 같았고, 이중에서 온도와 화합물 요인을 적절히 활용할 경우 보다 유용한 갈파래 발생제어 효과를 거둘 수 있을 것으로 전망되었다. Green tides, which was mainly caused by Ulva spp., have been increasing in severity and frequency globally, and have negatively affected on marine ecosystems. This study was conducted to investigate effects of various physical and chemical factors on the death of Ulva australis (ULAUS) and to consider a practical measures useful for alleviating Ulva bloom. Soaking of ULAUS thalli in pure water for 8 hr didn't induce a death, but incubation in 1.0-1.5% salinity for 7 d inhibited sporulation by about 70%. Desiccation gave rise to a serious damage when more than 40-50% of initial fresh weight was lost. ULAUS growth was sensitive to temperature and seriously inhibited from more than $30^{\circ}C$. At $35^{\circ}C$, $40^{\circ}C$, $45^{\circ}C$ and $50^{\circ}C$, treatment time required for 90-95% death of ULAUS thalli was 1 d, 10 min, 30 sec, and 1 sec, repectively. ULAUS growth was seriously inhibited at lower than pH 6.0 and completely dead at pH 4.0. Several compounds for ULAUS control was selected and the chemcals causing a rapid death were oxidants such as hydrogen peroxide and sodium percarbonate. Taken together, our results suggest that low salinities, dryness, pH, high temp. and compounds could be selected for Ulva bloom control, and high temperature and compounds seems to be useful for a development of practical control methods.

Glyphosate에 대한 옥수수 반응의 개선된 검정방법

김진석,이병회,김소희,민석기,최정섭,Kim, Jin-Seog,Lee, Byung-Hoi,Kim, So-Hee,Min, Suk-Ki,Choi, Jung-Sup 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.1

본 연구에서는 옥수수를 대상으로 glyphosate에 대한 여러 가지 생리적 반응을 검토한 후, glyphosate 저항성 평가에 활용될 수 있는 보다 개선된 방법 두 가지를 확립하였다. 한 가지 방법은 옥수수 제3엽 상단에 약제를 국소 처리한 다음, 처리 후 3일째에 약제처리 되지 않았던 제4엽의 신장 정도를 조사하는 것이다 (전식물체-엽생장 검정). 이 경우 glyphosate $50-1,600{\mu}g/mL$ 범위에서 농도가 증가됨에 따라 엽 생장이 억제되었으며, $1,600{\mu}g/mL$ 농도에서의 생장 억제율은 무처리 대비 55.5%였다. 다른 한 가지 방법은 옥수수 제3본엽의 엽절편 ($4{\times}4mm$) 4개씩을 $200{\mu}L$의 시험용액이 담긴 48 well plate에 치상한 후 $25^{\circ}C$ 연속 명조건에 24시간동안 배양하여 shikimate 축적량을 조사하는 것이다 (엽절편-shikimate 축적 검정). 이 경우 시험용액에 0.33% sucrose를 가하면 무첨가에 비해 약3-4배 정도의 shikimate 축적 증가가 관찰되었고 glyphosate $2-8{\mu}g/mL$ 농도범위에서 직선적 증가반응을 나타내어 기존방법 (Shaner et al. 2005)보다 개선된 특징을 보였다. 본 방법들은 glyphosate 저항성 옥수수를 창출할 때 또는 저항성 유전자의 타 식물로의 이동여부와 잡초화된 저항성 옥수수 존재여부를 감별하는데 활용될 수 있을 것이다. 이 때 glyphosate에 대한 저항성 원인이 작용점 EPSPS와 관련이 있는 경우에는 "엽절편-shikimate 축적 검정"이 가장 바람직하고, 저항성 원인이 체내이행 감소 때문일 경우에는 "전식물체-엽생장 검정" 수행이 필요하다. Several methods for determining the response of corn to glyphosate were investigated to provide a fast and reliable method for identifying glyphosate-resistant corn in vivo. Two bioassays were developed. One assay is named 'whole plant / leaf growth assay', in which the herbicide glyphosate is applied on the upper part of 3rd leaf and the growth of herbicide-untreated 4th leaf is measured at 3 day after treatment. in this assay, the leaf growth of conventional corn was inhibited in a dose dependent from 50 to $1600{\mu}g/mL$ of glyphosate and growth inhibition at $1600{\mu}g/mL$ was 55% of untreated control. The assay has the potential to be used especially in the case that the primary cause of glyphosate resistance is related with a reduction of the herbicide translocation. Another assay is named 'leaf segment / shikimate accumulation assay', in which the four excised leaf segments ($4{\times}4mm$) are placed in each well of a 48-well microtiter plate containing $200{\mu}L$ test solution and the amount of shikimate is determined after incubation for 24 h in continuous light at $25^{\circ}C$. In this assay, 0.33% sucrose added to basic test solution enhanced a shikimate accumulation by 3 to 4 times and the shikimate accumulation was linearly occurred from 2 to $8{\mu}g/mL$ of glyphosate, showing an improved response to the method described by Shaner et al. (2005). The leaf segment / shikimate accumulation assay is simple and robust and has the potential to be used as a high throughput assay in the case that the primary cause of glyphosate resistance is related with EPSPS, target site of the herbicide. Taken together, these two assays would be highly useful to initially select the lines obtained after transformation, to investigate the migration of glyphosate-resistant gene into other weeds and to detect a weedy glyphosate-resistant corn in field.

수출형 KT-1 훈련기 고/저온 환경시험절차 적합화 연구

김진석,Kim, Jin Seog 한국시스템엔지니어링학회 2020 시스템엔지니어링학술지 Vol.16 No.2

The environmental tests are one of the key branches of the reliability tests. The purpose of the high/low environmental test of KT-1 Export Version Trainer is not only based on meeting the compliance of the reliability, customer satisfaction, but also for the expansion of its export. Amongst many existing environmental tests, the high and low temperature tests are considered as the two most important tests. The tests are considered and tailored to the high and low temperature methods of MIL-STD-810H. This paper proposes the tailoring processes and the results of High/Low temperature conditions, test procedures, and test profiles for the aircraft. Additionally, this paper is described by considering the view of MIL-STD-810H.

XKT-1T 항공기 환경 요구사항 적합화 연구

김진석,Kim, Jin Seog 한국시스템엔지니어링학회 2021 시스템엔지니어링학술지 Vol.17 No.1

The purpose of the study on the tailoring of environmental requirements for KT-1 Export Version(XKT-1T) aircraft is not only based on meeting the compliance of the environmental reliability, customer satisfaction, but also for the expansion of its export. This paper proposes the environmental requirements tailoring processes and High/Low temperature conditions, test procedures, and the results of test profiles for the aircraft. Amongst many existing environmental requirements, the temperature requirements are considered as the most basic and the most important environmental requirements. The High/Low temperature tests are considered and tailored to the high and low temperature methods of MIL-STD-810.

경엽처리 제초제의 작용기작별 엽록소 형광발산 특성

김진석(Jin-Seog Kim),이병회(Byung-Hoi Lee),오광훈(Kwang-Hoon Oh),이정애(Jung-Ae Lee) 韓國雜草學會 2004 Weed&Turfgrass Science Vol.24 No.2

Mevalocidin+Diuron과 Mevalocidin+Tiafenacil 혼합물의 경엽처리시 제초활성 발현특성

김진석(Jin-Seog Kim),김재덕(Jae Deok Kim),최정섭(Jung Sup Choi),김은애(Eun Ae Kim),고영관(Young Kwan Ko),이일영(Ill Young Lee) 한국잡초학회 2022 한국잡초학회 별책(학술대회 초록집) Vol.42 No.1

목초액으로부터 제초활성 성분의 분리 및 동정

김진석(Jin-Seog Kim),김진철(Jin-Cheol Kim),최정섭(Jung Sup Choi),김태준(Tae-Joon Kim),김성문(Songmun Kim),조광연(Kwang Yun Cho) 韓國雜草學會 2001 Weed&Turfgrass Science Vol.21 No.4

Glyphosate 처리된 여러 식물의 shikimate 축적반응 특징

김진석(Jin-Seog Kim),최정섭(Jung Sup Choi),서보람(Bo Ram Seo),황현진(Hyun Jin Hwang) 한국잡초학회 2008 한국잡초학회 별책(학술대회 초록집) Vol.28 No.2