Kang S.,Yonsei University |
Shim H.S.,Yonsei University |
Lee J.S.,Bruker |
Kim D.S.,Genomine Inc. |
And 5 more authors.
Journal of Proteome Research
The specific molecular profiles of ovarian cancer interface zones (IZ), the region between tumors and normal tissues, were evaluated using a new method involving matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS). We analyzed three ovarian serous carcinomas using MALDI-IMS. Principal component analysis (PCA) was used to evaluate the quality of tissue spatial features based on MALDI-IMS, and for analysis of large data sets of MALDI-IMS. Twodimensional gel electrophoresis and fluorescence microscopy were used to verify interface-specific proteins. Unique profiles were identified for the tumors, the normal zone, and the IZ. Through MALDI analysis, two interfacespecific proteins, plastin 2 and peroxiredoxin 1 (PRDX 1), were identified as differentially regulated between zones. Fluorescence microscopy revealed high expression levels of plastin 2 and PRDX 1 along the IZ of ovarian tumors. This comparative proteomics study using tissue MALDIIMS suggested that the IZ is different from the adjacent tumor and normal zones, and that plastin 2 and PRDX 1 may be interface markers specific to ovarian tumors. © 2010 American Chemical Society. Source
Kim H.J.,Dong - A University |
Kim M.-J.,Nongwoo Bio Co. |
Pak J.H.,Dong - A University |
Im H.H.,Dong - A University |
And 7 more authors.
Plant Biotechnology Reports
Soybean [Glycine max (L.) Merr.] is an important crop for vegetable oil production, and is a major protein source worldwide. Because of its importance as a crop, genetic transformation has been used extensively to improve its valuable traits. Soybean mosaic virus (SMV) is one of the most well-known viral diseases affecting soybean. Transgenic soybean plants with improved resistance to SMV were produced by introducing HC-Pro coding sequences within RNA interference (RNAi) inducing hairpin construct via Agrobacterium-mediated transformation. During an experiment to confirm the response of transgenic plants (T2) to SMV infection, no T2 plants from lines #2 (31/31), #5 (35/35) or #6 (37/37) exhibited any SMV symptoms, indicating strong viral resistance (R), whereas NT (non-transgenic wild type) plants and those from lines #1, #3 and #4 exhibited mild mosaic (mM) or mosaic (M) symptoms. The northern blot analysis showed that three resistant lines (#2, #5 and #6) did not show the detection of viral RNA accumulation while NT, EV (transformed with empty vector carrying only Bar) and lines #1, #3 and #4 plants were detected. T3 seeds from SMV-inoculated T2 plants were harvested and checked for changes in seed morphology due to viral infection. T3 seeds of lines #2, #5 and #6 were clear and seed coat mottling was not present, which is indicative of SMV resistance. RT-PCR and quantitative real-time PCR showed that T3 seeds from the SMV-resistant lines #2, #5 and #6 did not exhibit any detection of viral RNA accumulation (HC-Pro, CP and CI), while the viral RNA accumulation was detected in SMV-susceptible lines #1, #3 and #4 plants. During the greenhouse test for viral resistance and yield components, T3 plants from the SMV-inoculated transgenic lines #2, #5 and #6 showed viral resistance (R) and exhibited a more favorable average plant height, number of nodes per plant, number of branches per plant, number of pods per plant and total seed weight with statistical significance during strong artificial SMV infection than did other plant lines. In particular, the SMV-resistant line #2 exhibited superior average plant height, pod number and total seed weight with highly significance. According to our results, RNAi induced by the hairpin construct of the SMV HC-Pro sequence effectively confers much stronger viral resistance than did the methods used during previous trials, and has the potential to increase yields significantly. Because of its efficiency, the induction of RNAi-mediated resistance will likely be used more frequently as part of the genetic engineering of plants for crop improvement. © 2016 Korean Society for Plant Biotechnology and Springer Japan Source
Hwang I.-T.,Korea Research Institute of Chemical Technology |
Choi J.-S.,Korea Research Institute of Chemical Technology |
Song H.-Y.,Korea Research Institute of Chemical Technology |
Cho S.-J.,Korea Research Institute of Chemical Technology |
And 3 more authors.
Pesticide Biochemistry and Physiology
The validation of potential herbicide target, 7-keto-8-aminopelargonic acid synthase (KAPAS) in the early step of biotin biosynthesis pathway, was performed in vitro and in vivo with lead chemical triphenyltin acetate (TPTA). KAPAS activity was completely inhibited by TPTA with an IC50 of 19.85 μM. 40-day-old Arabidopsis thaliana plants were killed with foliar treatment of 125 g ha-1 TPTA under the greenhouse conditions. The germination of A. thaliana seeds was also completely inhibited with 62.5 μM TPTA, but it was rescued to 85-92% with the supplement of biotin biosynthesis intermediates such as 0.5 mM of biotin, dethiobiotin, and 7,8-diaminopelargonic acid, but not by 7-keto-8-aminopelargonic acid (KAPA). However, additional supplement of 0.5 mM S-adenosyl-l-methionine (SAM) with 0.5 mM KAPA rescued up to 91% of the germination previously inhibited by the 50 μM TPTA. Also, biotin supplements alleviated the growth inhibition of 40-day-old A. thaliana plant. Foliar application of TPTA induced 8-fold higher substrate (l-alanine) accumulation in the treated A. thaliana plants. RNA expression for KAPAS transcripts were much fainter in the lane representing leaf tissue treated with TPTA. With these results, we report that SAM is an essential donor of amino groups for synthesis of the biotin precursor KAPA to 7,8-diaminopelargonic acid (DAPA) synthesis in plants, that KAPAS is a potential herbicidal target site in the biotin biosynthesis pathway, and that TPTA might be one of the potential KAPAS inhibitors. © 2009 Elsevier Inc. All rights reserved. Source
Lee D.H.,Genomine Inc. |
Lee I.C.,Daegu Gyeongbuk Institute of Science and Technology |
Kim K.J.,Genomine Inc. |
Kim D.S.,Genomine Inc. |
And 6 more authors.
Journal of Plant Biology
Gibberellin (GA), a plant hormone, is involved in many aspects of plant growth and development both in vegetative and reproductive phases. GA2-oxidase plays a key role in the GA catabolic pathway to reduce bioactive GAs. We produced transgenic Arabidopsis plants expressing GA2-oxidase 4 (AtGA2ox4) under the control of a senescenceassociated promoter (SEN1). As we hypothesized, transgenic plants (SEN1::AtGA2ox4) exhibited a dominant semi-dwarf phenotype with a decrease of bioactive GAs (e.g., GA4 and GA1) up to two-fold compared to control plants. Application of bioactive GA3 resulted in increased shoot length, indicating that the GA signaling pathway functions normally in the SEN1::AtGA2ox4 plants. Expressions of other members of GA2-oxidase family, such as AtGA2ox1, AtGA2ox3, AtGA2ox6, and AtGA2ox8, were decreased slightly in the flower and silique tissues while GA biosynthetic genes (e.g., AtGA20ox1, AtGA20ox2 and AtGA3ox1) were not significantly changed in the SEN::AtGA2ox4 plants. Using proteome profiling (2-D PAGE followed by MALDI-TOF/MS), we identified 29 protein spots that were increased in the SEN1::AtGA2ox4 plants, but were decreased to wild-type levels by GA3 treatment. The majority were found to be involved in photosynthesis and carbon/energy metabolism. Unlike the previous constitutive over-expression of GA2-oxidases, which frequently led to floral deformity and/or loss of fertility, the SEN1::AtGA2ox4 plants retained normal floral morphology and seed production. Accordingly, the expressions of FT and CO genes remained unchanged in the SEN1::AtGA2ox4 plants. Taken together, our results suggest that the dominant dwarf trait carried by SEN1::AtGA2ox4 plants can be used as an efficient dwarfing tool in plant biotechnological applications. © 2014 Korean Society of Plant Biologists and Springer-Verlag Berlin Heidelberg. Source
Choi H.,Korea Basic Science Institute |
Choi H.,Pohang University of Science and Technology |
Jeong S.,Korea Basic Science Institute |
Jeong S.,Pohang University of Science and Technology |
And 10 more authors.
Phytochromes are red (R)/far-red (FR) photoreceptors that are central to the regulation of plant growth and development. Although it is well known that photoactivated phytochromes are translocated into the nucleus where they interact with a variety of nuclear proteins and ultimately regulate genome-wide transcription, the mechanisms by which these photoreceptors function are not completely understood. In an effort to enhance our understanding of phytochrome-mediated light signaling networks, we attempted to identify novel proteins interacting with phytochrome B (phyB). Using affinity purification in Arabidopsis phyB overexpressor, coupled with mass spectrometry analysis, 16 proteins that interact with phyB in vivo were identified. Interactions between phyB and six putative phyB-interacting proteins were confirmed by bimolecular fluorescence complementation (BiFC) analysis. Involvement of these proteins in phyB-mediated signaling pathways was also revealed by physiological analysis of the mutants defective in each phyB-interacting protein. We further characterized the athb23 mutant impaired in the homeobox protein 23 (ATHB23) gene. The athb23 mutant displayed altered hypocotyl growth under R light, as well as defects in phyB-dependent seed germination and phyB-mediated cotyledon expansion. Taken together, these results suggest that the ATHB23 transcription factor is a novel component of the phyB-mediated R light signaling pathway. © 2013 Scandinavian Plant Physiology Society. Source