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수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정
신평균(Pyung-Gyun Shin),장안철(Ancheol Chang),홍성창(Sung-Chang Hong),이기상(Ki-Sang Lee) 한국생명과학회 2007 생명과학회지 Vol.17 No.12
BAK1(Brassinolide insensitive associated receptor kinase 1)는 브라시노스테로이드 생합성 대사관련 신호전달 매체이다. BR 생합성 및 신호전달 돌연변이체는 매우 특징적인 난쟁이 표현형을 보인다. 과채류전용 양액배지인 Sonneveld 양액을 이용하여 양분결핍에 의해 왜성을 나타내는 토마토를 선발하였다. 선발된 토마토에 대해 이차원 전기영동법으로 단백질체를 분석한 결과, 발현차를 나타내는 28개의 단백질 spot이 분리되었다. 분리된 단백질 spot 중 현저하게 발현이 억제된 단백질 spot 6개를 선발하여 단백질 서열을 결정하였다. 실험결과, pI 4.5, 분자량 24 kDa를 나타내는 단백질은 브라시노스테로이드 생합성에 관여하며 왜성 표현형을 나타내는 신호전달 단백질, BAK1으로 동정되었다. BCK1, cystein proteinase, sulfutase, peroxidase, zinc finger factor로 동정된 나머지 단백질들은 브레시노스테로이드 생합성관련 신호전달기작에 관여하는 단백질로 추정되었다. BAK1을 검정하기 위해 단백질 서열이 결정된 부위로부터 프라이머를 디자인하여 RT-PCR를 수행한 결과, 증폭된 500 bp의 산물이 정상과 발현차를 보여주었는데 이 결과는 양분조절에 의해서도 BAK1의 발현이 조절될 수 있음을 시사한다. Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.
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Proteomics를 이용한 고랭지 배추의 고온장해 해석
신평균(Pyung-Gyun Shin),홍성창(Sung-Chang Hong),장안철(Ancheol Chang),김상효(Sang-Hyo Kim),이기상(Ki-Sang Lee) 한국생명과학회 2007 생명과학회지 Vol.17 No.12
무더운 날씨가 지속됨으로서 고랭지배추의 생장 및 결구가 지연되고 있는 강원도 정선군 질운산(새빗재)의 600 m와 900m의 배추를 사용하여 무기성분 및 단백질 발현패턴을 분석하였다. 식물체 무기성분에서는 생장에 관련된 질소 및 인산의 부족현상과 결구에 관련된 칼슘이 부족하였다. 단백체 분석은 2차원 전기영동에 의해 전체 126개의 단백질이 분리되었고 그 중 48개의 단백질이 고도에 따라 변화하는 양상을 보여주었다. 이 중에서 30개의 단백질 서열이 결정되었는데, 해발 900 m에서 단백질 발현이 증가한 14개 중에서 oxygen- evolving proteins, rubisco activase and ATPase 등이, 해발 600 m에서는 glutathione S-transferase (1, 28 kD cold induced- and 24 kD auxin-binding proteins) and salt-stress induced protein 등 16개의 단백질 발현이 증가하였다. 이러한 단백질은 식물체 손상에 대한 보호기작을 가진 스트레스관련 단백질로 가뭄, 온도상승, 밤낮의 온도차 등의 반복으로 복합적이며 동시 다발적으로 나타나는 고온장해 현상으로 사료된다. High temperature stresses have caused growth inhibition and delayed heading in highland cultivation Chinese cabbage during summer in Korea. We have studied high temperature stress responses in the terms of changes of inorganic components and proteins by proteomic analyses. Insufficiencies of nitrogen and phosphorus have affected growth rate and calcium deficiency has caused blunted heading. Proteins extracted from Brassica seedling grown at the altitude of 600m and 900m in the Mount Jilun were extracted and analysed by 2-dimentional polyacrylamide gel electrophoresis. Profiles of protein expression was then analyzed by 2-dimentional gel analyses. Protein spots showing different expression level were picked using the spot handling workstation and subjected to MALDI-TOF MS. Total 48 protein spots were analyzed by MALDI-TOF MS and 30 proteins spots out of 48 were identified by peptide mass fingerprinting analyses. Fourteen proteins were up-regulated in extracts from the altitude of 900m and they were identified as oxygen-evolving proteins, rubisco activase and ATPase etc. Sixteen proteins were up-regulated in extracts from the altitude of 600m and they were identified as glutathione S-transferase(1, 28 kD cold induced- and 24 kD auxin-binding proteins) and salt-stress induced protein etc. These stress-induced proteins were related to the mediated protective mechanism against oxidative damage during various stresses. The results indicated that physiological phenomenon in response to high temperature stresses might be resulted by complex and multiple array of responses with drought, heat, oxidative, salt, and cold by high temperature.
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손수인 ( Soo-in Sohn ),오영주 ( Young-ju Oh ),김병용 ( Byung-yong Kim ),장안철 ( Ancheol Chang ),이범규 ( Bumkyu Lee ),윤도원 ( Doh-won Yun ),이강섭 ( Gang-seob Lee ),오성덕 ( Sung-dug Oh ),조현석 ( Hyun-suk Cho ) 한국환경농학회 2016 한국환경농학회 학술대회집 Vol.2016 No.-
The studies about the effect of planted GM crops on soil microorganisms have been continued. Although many studies have been carried out over the past decades, they provided contradictory information about the effect on soil microorganisms even for the same GM crop due to the diversity of soil environment, which means the effect of GM crops on soil microorganisms should be considered in many aspects. In this study, we investigated the effect of the drought tolerant rice, MSRB2-Bar-8 which expresses CaMSRB2 gene on soil microorganisms based on the culture-dependent and culture-independent methods. For this, soil chemical analysis, population density of soil microorganisms, denaturing gradient gel electrophoresis (DGGE) and pyrosequencing were analyzed with rhizosphere soils of GM and the non-GM counterpart of Ilmi rice. There was no significant difference in soil chemicals between GM and non-GM rice. The microbial community densities of the GM soils were found to be within the range of those of the non-GM rice. The DGGE patterns of the total soil microorganisms for GM and non-GM rice were also similar. In the pyrosequencing analyses, Proteobacteria and Cloroflexi were dominant at seedling stage while Cloroflexi showed dominance over Proteobacteria at maturity stage in both soils. In UPGMA dendrogram, two soils were grouped by time period showing difference in soil microorganisms by time period; however, such difference was not the difference in soil microorganisms between GM and non-GM. In conclusion, the results from this study imply that the effect of MSRB2-Bar-8 cultivation on soil microorganisms is not significant.
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