• Title/Summary/Keyword: Seeds localization

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Seeds as Repositories of Recombinant Proteins in Molecular Farming

  • Moloney, Maurice M.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.283-297
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    • 2000
  • Seeds are an ideal repository for recombinant proteins in molecular farming applications. However, in order to use plant seeds efficiently for the production of such proteins, it is necessary to understand a number of fundamental biological properties of seeds. This includes a full understanding of promoters which function in a seed-specific manner, the subcellular targeting of the desired polypeptide and the final form in which a protein is stored. Once a biologically active protein has been deposited in a seed, it is also critical that the protein can be extracted and purified efficiently. In this review, these issues are examined critically to provide a number of approaches which may be adopted for production of recombinant proteins in plants. Particular attention is paid to the relationship between subcellular localization and protein extraction and purification. The robustness and flexibility of seed-based production is illustrated by examples close to or already in commercial production.

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A Study on Image Reconstruction for Seed Localization for Permanent Prostate Brachytherapy (전립선암 근접치료 시 방사성선원 위치확인을 위한 영상 재구성에 관한 연구)

  • Hong, Ju-Young;Rah, Jeong-Eun;Suh, Tae-Suk
    • Radiation Oncology Journal
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    • v.25 no.2
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    • pp.125-133
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    • 2007
  • [ $\underline{Purpose}$ ]: This study was to design and fabricate a phantom for prostate cancer brachytherapy to validate a developed program applying a 3-film technique, and to compare it with the conventional 2-film technique for determining the location of an implanted seed. $\underline{Materials\;and\;Methods}$: The images were obtained from overlapped seeds by randomly placing a maximum of 63 seeds in the anterior-posterior (AP) position and at $-30^{\circ} to $30^{\circ} at $15^{\circ} intervals. Images obtained by use of the phantom were applied to the image processing procedure, and were then processed into the development program for seed localization. In this study, cases were set where one seed overlapped, where two seeds overlapped and where none of the three views resolved all seeds. The distance between the centers of each seed to the reference seed was calculated in a prescribed region. This distance determined the location of each seed in a given band. The location of the overlapped seeds was compared with that of the 2-film technique. $\underline{Results}$: With this program, the detection rate was 92.2% (at ${\pm}15^{\circ}), 94.1% (at ${\pm}30^{\circ}) and 70.6% (compared to the use of the 2-film technique). The overlaps were caused by one or more than two seeds that overlapped; the developed program can identify the location of each seed perfectly. However, for the third case the program was not able to resolve the overlap of the seeds. $\underline{Conclusion}$: This program can be used to improve treatment outcome for the brachytherapy of prostate cancer by reducing the number of errors in the process of reconstructing the locations of perfectly overlapped seeds.

Cone-beam Reconstruction using Limited EPID Projections for Seeds Localization (Seed의 위치 확인을 위한 제한된 EPID 영상을 이용한 콘빔 재구성)

  • Chang, Ji-Na;Jung, Won-Kyun;Park, Sung-Ho;Cheong, Kwang-Ho;Suh, Tae-Suk
    • Progress in Medical Physics
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    • v.19 no.3
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    • pp.186-190
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    • 2008
  • In this study, we describe the preliminary application for the delineation of a metal object using cone-beam reconstruction (CBR) based on limited electronic portal imaging device (EPID) projections. A typical Feldkamp, Davis and Kress (FDK) reconstruction algorithm accompanying the edge preserving smoothing filter was used as only a few projections are acquired for reconstruction. In a correlation study of the projection numbers, we found that the size of the seeds and their location depicted by these CBR images were almost identical. Limited views were used for CBR, and our method is inexpensive and competitive for use in clinical applications.

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Localization and Function of Cellulase in Endosperm Cells of Panax ginseng Seeds during Maturation and After-ripening (인삼 종자의 성숙과 후숙 과정에서 배유세포내 섬유소 가수분해효소의 분포 및 기능)

  • 유성철
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.327-335
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    • 1993
  • The active sites, intracellular transport, function of cellulase in association with the disintegration of the storage materials of the endosperm cells during seed maturation and after-ripening of Panax ginseng C.A. Meyer seeds were studied by electron microscopy. Cytochemical activities of the cellulase occurred in protein bodies and vesicles of endosperm cells in seed with red seed coat. In after-ripening seed, the activities were strongly found in the cell wall of endosperm near the umbiliform layer and on neighbouring vesicles, so it is assumed that these cells begin to be decomposed. Cellulase activities were initiated before the decomposition of storage materials. But, no activity was observed in the umbiliform layer, so it is suggested that cellulase lose its activity after the completion of lysis process.

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Distribution of Lipid and Lipase in Lipid-and Starch-Rich Seeds (지질 및 전분성 종자에서 지질 및 지질가수분해효소의 분포)

  • 김우갑
    • Journal of Plant Biology
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    • v.35 no.3
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    • pp.219-227
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    • 1992
  • Formation, cellular distribution and structural changes of storage lipid, and active site and cellular localization of lipase in endosperms and cotyledons of lipid-rich seeds such as Helianthus annuus, Ricinus communis and Pinus koraiensis, and in those of starch-rich seeds such as Pisum sativum and Zea mays were investigated in relation to the seed development by cytochemical methods. In endosperms and storage cotyledons of lipid- and starch-rich seeds after seed-gathering, there were widely distributed storage material which was composed of spherical protein bodies, spherosomes, and starch granules. But cellular organelles were hardly observed in the cytoplasm. Staining pattern of vesicles released from SER, and of low electron dense membraneous granules, which were perhaps at an early stage of spherosomes, were the same as in the spherosome. Electrondense granules released from RER were observed in the vicinity of plasma membrane. As a result of lipid staining, the spherosomes were more electron dense and were uniform as compared with the protein matrix within the protein body and cytoplasmic proteinaceous granules. The major component of the spherosome was determinated to be lipid. Spherosomes and vesicles containing SER-released materials showed the same as in the electron density. Lipase activity was especially strong in the inner region and on the surface of decomposed spherosomes and near the plasma membrane.mbrane.

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Pseudomonas oleovorans Strain KBPF-004 Culture Supernatants Reduced Seed Transmission of Cucumber green mottle mosaic virus and Pepper mild mottle virus, and Remodeled Aggregation of 126 kDa and Subcellular Localization of Movement Protein of Pepper mild mottle virus

  • Kim, Nam-Gyu;Seo, Eun-Young;Han, Sang-Hyuk;Gong, Jun-Su;Park, Cheol-Nam;Park, Ho-Seop;Domier, Leslie L;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.393-401
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    • 2017
  • Efforts to control viral diseases in crop production include several types of physical or chemical treatments; antiviral extracts of a number of plants have also been examined to inhibit plant viral infection. However, treatments utilizing naturally selected microorganisms with activity against plant viruses are poorly documented. Here we report isolation of a soil inhabiting bacterium, Pseudomonas oleovorans strain KBPF-004 (developmental code KNF2016) which showed antiviral activity against mechanical transmission of tobamoviruses. Antiviral activity was also evaluated in seed transmission of two tobamoviruses, Pepper mild mottle virus (PMMoV) and Cucumber green mottle mosaic virus (CGMMV), by treatment of seed collected from infected pepper and watermelon, respectively. Pepper and watermelon seeds were treated with culture supernatant of P. oleovorans strain KBPF-004 or control strain ATCC 8062 before planting. Seeds germinated after treatment with water or ATCC 8062 yielded about 60% CGMMV or PMMoV positive plants, whereas < 20% of KBPF-004-treated seeds were virus-infected, a significantly reduced seed transmission rate. Furthermore, supernatant of P. oleovorans strain KBPF-004 remodeled aggregation of PMMoV 126 kDa protein and subcellular localization of movement protein in Nicotiana benthamiana, diminishing aggregation of the 126 kDa protein and essentially abolishing association of the movement protein with the microtubule network. In leaves agroinfiltrated with constructs expressing the coat protein (CP) of either PMMoV or CGMMV, less full-size CP was detected in the presence of supernatant of P. oleovorans strain KBPF-004. These changes may contribute to the antiviral effects of P. oleovorans strain KBPF-004.

OsDOR1, a novel glycine rich protein that regulates rice seed dormancy

  • Kim, Suyeon;Huh, Sun Mi;Han, Hay Ju;Cho, Mi Hyun;Lee, Gang Sub;Kim, Beom Gi;Kwon, Taek Yun;Yoon, In Sun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.90-90
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    • 2017
  • Regulation of seed dormancy is important in many grains to prevent pre-harvest sprouting. To identify and understand the gene related to seed dormancy regulation, we have screened for viviparous phenotypes of rice mutant lines generated by insertion of Ds transposon in a Korean Japonica cultivar (Dongjin) background. One of the mutants, which represented viviparous phenotype, was selected for further seed dormancy regulation studies and designated dor1. The dor1 mutant has single Ds insertion in the second exon of OsDor1 gene encoding glycine-rich protein. The seeds of dor1 mutant showed a higher germination potential and reduced abscisic acid (ABA) sensitivity compared to wild type Dongjin. Over-expression of Dor1 complements the viviparous phenotype of dor1 mutant, indicating that Dor1 function in seed dormancy regulation. Subcellular localization assay of Dor1-GFP fusion protein revealed that the OsDor1 protein mainly localized to membrane and the localization of OsDOR1 was influenced by presence of a giberelin (GA) receptor OsGID1. Further bimolecular fluorescence complementation (BiFC) analysis indicated that OsDOR1 interact with OsGID1. The combined results suggested that OsDOR1 regulates seed dormancy by interacting with OsGID1 in GA response. Additionally, expression of OsDOR1 partially complemented the cold sensitivity of Escherichia coli BX04 mutant lacking four cold shock proteins, indicating that OsDOR1 possessed RNA chaperone activity.

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Localization of Lipoxygenase in Germinating Soybeans (발아중인 대두콩에서의 Lipoxygenase의 국재(局在))

  • Song, Young-Sun
    • Korean Journal of Food Science and Technology
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    • v.19 no.5
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    • pp.441-445
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    • 1987
  • The subcellular distribution of lipoxygenase in germinating soybean seeds (Glycine max[L.] AmSoy) was investigated by using differential centrifugation and sucrose density gradient fractionation. Most of lipoxygenase -1 and -2/3 activities was present in the supernatant fraction after differential centrifugation of homogenates prepared from three-day-old seedlings; only 1.5% of lipoxygenase activity remained in particulate fractions. The results of a sucrose density gradient fractionation (three-day-old) showed that the lipoxygenase activity coincided with acid phosphatase at the densities of 1.19, 1.23, $1.25g/cm^3$, even though most of lipoxygenase and acid phosphatase activities appeared in supernatant fractions. There was no indication that mitochondria contained any lipoxygenase activity, and it does not appear that glyoxysomes and ER contained any lipoxygenase activity either.

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EST Profiling for Seed-hair Characteristic and Development of EST-SSR and SNP Markers in Carrot (당근 종모 형질 관련 EST profiling과 이를 이용한 EST-SSR 및 SNP 마커 개발)

  • Oh, Gyu-Dong;Hwang, Eun-Mi;Shim, Eun-Jo;Jeon, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1025-1038
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    • 2010
  • Carrot ($Daucus$ $carota$ L. var. $sativa$) is one of the most widely used crops in the world. Moreover it is an important crop because of its high content of ${\beta}$-carotene, well-known as the precursor of vitamin A carotenoid. However, seed-hair which is generated in epidermal cell of seeds inhibits absorption and germination. For that reason, carrot seeds are commercialized after mechanical hair removal process. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-ATR615 OP 666-13line and hairy seed CT-ATR615 OP-CK1-9 line were constructed and expression patterns related to generation of seed-hair were analyzed by comparison of EST sequences. Differential EST sequence results between two lines were classified into FunCat functional categories based on the results of BlastX search. Higher expression quantities belonging to metabolic category were shown on short-hair seed line than hairy-seed one. Differential expression quantities between those two lines in the protein folding and stabilization, subcellular localization categories were supposed to contribute variously on the generation of seed-hair. We confirmed 50 and 59 SSR sites, and 2 SNP sites by analyzing EST sequences in two lines; thereafter, we designed SNP and SSR primer sets from these EST sequence information as a molecular marker. These markers are thought to be used in research of molecular markers for classification of carrot family and related to various traits, as well as seed-hair characteristic.