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[PMID]:29253877
[Au] Autor:Geldreich A; Haas G; Kubina J; Bouton C; Tanguy M; Erhardt M; Keller M; Ryabova L; Dimitrova M
[Ad] Endereço:Institut de Biologie Moléculaire des Plantes, CNRS UPR2357, Université de Strasbourg, Strasbourg, France.
[Ti] Título:Formation of large viroplasms and virulence of Cauliflower mosaic virus in turnip plants depend on the N-terminal EKI sequence of viral protein TAV.
[So] Source:PLoS One;12(12):e0189062, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cauliflower mosaic virus (CaMV) TAV protein (TransActivator/Viroplasmin) plays a pivotal role during the infection cycle since it activates translation reinitiation of viral polycistronic RNAs and suppresses RNA silencing. It is also the major component of cytoplasmic electron-dense inclusion bodies (EDIBs) called viroplasms that are particularly evident in cells infected by the virulent CaMV Cabb B-JI isolate. These EDIBs are considered as virion factories, vehicles for CaMV intracellular movement and reservoirs for CaMV transmission by aphids. In this study, focused on different TAV mutants in vivo, we demonstrate that three physically separated domains collectively participate to the formation of large EDIBs: the N-terminal EKI motif, a sequence of the MAV domain involved in translation reinitiation and a C-terminal region encompassing the zinc finger. Surprisingly, EKI mutant TAVm3, corresponding to a substitution of the EKI motif at amino acids 11-13 by three alanines (AAA), which completely abolished the formation of large viroplasms, was not lethal for CaMV but highly reduced its virulence without affecting the rate of systemic infection. Expression of TAVm3 in a viral context led to formation of small irregularly shaped inclusion bodies, mild symptoms and low levels of viral DNA and particles accumulation, despite the production of significant amounts of mature capsid proteins. Unexpectedly, for CaMV-TAVm3 the formation of viral P2-containing electron-light inclusion body (ELIB), which is essential for CaMV aphid transmission, was also altered, thus suggesting an indirect role of the EKI tripeptide in CaMV plant-to-plant propagation. This important functional contribution of the EKI motif in CaMV biology can explain the strict conservation of this motif in the TAV sequences of all CaMV isolates.
[Mh] Termos MeSH primário: Brassica napus/virologia
Caulimovirus/metabolismo
Caulimovirus/patogenicidade
Transativadores/química
Transativadores/metabolismo
[Mh] Termos MeSH secundário: Motivos de Aminoácidos
Sequência de Aminoácidos
Caulimovirus/ultraestrutura
Corpos de Inclusão Viral/metabolismo
Corpos de Inclusão Viral/ultraestrutura
Proteínas Mutantes/metabolismo
Fenótipo
Domínios Proteicos
Protoplastos/metabolismo
Transcrição Reversa/genética
Relação Estrutura-Atividade
Virulência
Replicação Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Mutant Proteins); 0 (Trans-Activators); 0 (gene VI protein, Cauliflower mosaic virus)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180108
[Lr] Data última revisão:
180108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171219
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189062


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[PMID]:28941403
[Au] Autor:Hyodo K; Nagai H; Okuno T
[Ad] Endereço:Institute of Plant Science and Resources, Okayama University, Kurashiki, Okayama 710-0046, Japan. Electronic address: khyodo@okayama-u.ac.jp.
[Ti] Título:Dual function of a cis-acting RNA element that acts as a replication enhancer and a translation repressor in a plant positive-stranded RNA virus.
[So] Source:Virology;512:74-82, 2017 Dec.
[Is] ISSN:1096-0341
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The genome of red clover necrotic mosaic virus is divided into two positive-stranded RNA molecules of RNA1 and RNA2, which have no 5' cap structure and no 3' poly(A) tail. Previously, we showed that any mutations in the cis-acting RNA replication elements of RNA2 abolished its cap-independent translational activity, suggesting a strong link between RNA replication and translation. Here, we investigated the functions of the 5' untranslated region (UTR) of RNA2 and revealed that the basal stem-structure (5'BS) predicted in the 5' UTR is essential for robust RNA replication. Interestingly, RNA2 mutants with substitution or deletion in the right side of the 5'BS showed strong translational activity, despite their impaired replication competency. Furthermore, nucleotide sequences other than the 5'BS of the 5' UTR were essential to facilitate the replication-associated translation. Overall, these cis-acting RNA elements seem to coordinately regulate the balance between RNA replication and replication-associated translation.
[Mh] Termos MeSH primário: Regulação Viral da Expressão Gênica/fisiologia
Tombusviridae/genética
Tombusviridae/fisiologia
Replicação Viral/fisiologia
[Mh] Termos MeSH secundário: Biossíntese de Proteínas
Protoplastos
RNA Viral/genética
Tabaco
Regiões não Traduzidas/genética
Regiões não Traduzidas/fisiologia
Proteínas Virais
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Untranslated Regions); 0 (Viral Proteins)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171109
[Lr] Data última revisão:
171109
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170924
[St] Status:MEDLINE


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[PMID]:28829800
[Au] Autor:Xu X; Xu X; Zhou Y; Zeng S; Kong W
[Ad] Endereço:School of Horticulture and Plant Protection, Yangzhou University, Yangzhou, China.
[Ti] Título:Identification of protoplast-isolation responsive microRNAs in Citrus reticulata Blanco by high-throughput sequencing.
[So] Source:PLoS One;12(8):e0183524, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Protoplast isolation is a stress-inducing process, during which a variety of physiological and molecular alterations take place. Such stress response affects the expression of totipotency of cultured protoplasts. MicroRNAs (miRNAs) play important roles in plant growth, development and stress responses. However, the underlying mechanism of miRNAs involved in the protoplast totipotency remains unclear. In this study, high-throughput sequencing technology was used to sequence two populations of small RNA from calli and callus-derived protoplasts in Citrus reticulata Blanco. A total of 67 known miRNAs from 35 families and 277 novel miRNAs were identified. Among these miRNAs, 18 known miRNAs and 64 novel miRNAs were identified by differentially expressed miRNAs (DEMs) analysis. The expression patterns of the eight DEMs were verified by qRT-PCR. Target prediction showed most targets of the miRNAs were transcription factors. The expression levels of half targets showed a negative correlation to those of the miRNAs. Furthermore, the physiological analysis showed high levels of antioxidant activities in isolated protoplasts. In short, our results indicated that miRNAs may play important roles in protoplast-isolation response.
[Mh] Termos MeSH primário: Citrus/genética
Sequenciamento de Nucleotídeos em Larga Escala/métodos
MicroRNAs/genética
Protoplastos
RNA de Plantas/genética
[Mh] Termos MeSH secundário: Citrus/metabolismo
Peróxido de Hidrogênio/metabolismo
Malondialdeído/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
0 (MicroRNAs); 0 (RNA, Plant); 4Y8F71G49Q (Malondialdehyde); BBX060AN9V (Hydrogen Peroxide)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171013
[Lr] Data última revisão:
171013
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170823
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183524


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[PMID]:28637336
[Au] Autor:Hayashi N; Sasaki S; Takahashi H; Yamashita Y; Naito S; Onouchi H
[Ad] Endereço:Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
[Ti] Título:Identification of Arabidopsis thaliana upstream open reading frames encoding peptide sequences that cause ribosomal arrest.
[So] Source:Nucleic Acids Res;45(15):8844-8858, 2017 Sep 06.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Specific sequences of certain nascent peptides cause programmed ribosomal arrest during mRNA translation to control gene expression. In eukaryotes, most known regulatory arrest peptides are encoded by upstream open reading frames (uORFs) present in the 5'-untranslated region of mRNAs. However, to date, a limited number of eukaryotic uORFs encoding arrest peptides have been reported. Here, we searched for arrest peptide-encoding uORFs among Arabidopsis thaliana uORFs with evolutionarily conserved peptide sequences. Analysis of in vitro translation products of 22 conserved uORFs identified three novel uORFs causing ribosomal arrest in a peptide sequence-dependent manner. Stop codon-scanning mutagenesis, in which the effect of changing the uORF stop codon position on the ribosomal arrest was examined, and toeprint analysis revealed that two of the three uORFs cause ribosomal arrest during translation elongation, whereas the other one causes ribosomal arrest during translation termination. Transient expression assays showed that the newly identified arrest-causing uORFs exerted a strong sequence-dependent repressive effect on the expression of the downstream reporter gene in A. thaliana protoplasts. These results suggest that the peptide sequences of the three uORFs identified in this study cause ribosomal arrest in the uORFs, thereby repressing the expression of proteins encoded by the main ORFs.
[Mh] Termos MeSH primário: Proteínas de Arabidopsis/genética
Arabidopsis/genética
Regulação da Expressão Gênica de Plantas
Fases de Leitura Aberta
Elongação Traducional da Cadeia Peptídica
Terminação Traducional da Cadeia Peptídica
[Mh] Termos MeSH secundário: Regiões 5' não Traduzidas
Sequência de Aminoácidos
Arabidopsis/metabolismo
Proteínas de Arabidopsis/metabolismo
Códon de Terminação
Sequência Conservada
Genes Reporter
Luciferases/genética
Luciferases/metabolismo
Protoplastos/metabolismo
Ribossomos/genética
Ribossomos/metabolismo
Alinhamento de Sequência
Homologia de Sequência de Aminoácidos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5' Untranslated Regions); 0 (Arabidopsis Proteins); 0 (Codon, Terminator); EC 1.13.12.- (Luciferases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171012
[Lr] Data última revisão:
171012
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170623
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx528


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[PMID]:28409545
[Au] Autor:Wacker MA; Teghanemt A; Weiss JP; Barker JH
[Ad] Endereço:1 Department of Biology, Central Michigan University, Mt. Pleasant, MI, USA.
[Ti] Título:High-affinity caspase-4 binding to LPS presented as high molecular mass aggregates or in outer membrane vesicles.
[So] Source:Innate Immun;23(4):336-344, 2017 May.
[Is] ISSN:1753-4267
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Caspases of the non-canonical inflammasome (caspases -4, -5, and -11) directly bind endotoxin (LOS/LPS) and can be activated in the absence of any co-factors. Models of LPS-induced caspase activation have postulated that 1:1 binding of endotoxin monomers to caspase trigger caspase oligomerization and activation, analogous to that established for endotoxin-induced activation of MD-2/TLR4. However, using metabolically radiolabeled LOS and LPS, we now show high affinity and selective binding of caspase-4 to high molecular mass aggregates of purified endotoxin and to endotoxin-rich outer membrane vesicles without formation of 1:1 endotoxin:caspase complexes. Thus, our findings demonstrate markedly different endotoxin recognition properties of caspase-4 from that of MD-2/TLR4 and strongly suggest that activation of caspase-4 (and presumably caspase-5 and caspase-11) are mediated by interactions with activating endotoxin-rich membrane interfaces rather than by endotoxin monomers.
[Mh] Termos MeSH primário: Caspases Iniciadoras/metabolismo
Vesículas Citoplasmáticas/metabolismo
Lipopolissacarídeos/metabolismo
Membranas Mitocondriais/metabolismo
Neisseria meningitidis/imunologia
Protoplastos/metabolismo
Staphylococcus aureus/imunologia
[Mh] Termos MeSH secundário: Caspases Iniciadoras/genética
Parede Celular/metabolismo
Seres Humanos
Ligação Proteica
Multimerização Proteica
Proteínas Recombinantes/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Lipopolysaccharides); 0 (Recombinant Proteins); EC 3.4.22.- (CASP4 protein, human); EC 3.4.22.- (Caspases, Initiator)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170804
[Lr] Data última revisão:
170804
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170415
[St] Status:MEDLINE
[do] DOI:10.1177/1753425917695446


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[PMID]:28323890
[Au] Autor:Huo A; Chen Z; Wang P; Yang L; Wang G; Wang D; Liao S; Cheng T; Chen J; Shi J
[Ad] Endereço:Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, China.
[Ti] Título:Establishment of transient gene expression systems in protoplasts from Liriodendron hybrid mesophyll cells.
[So] Source:PLoS One;12(3):e0172475, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Liriodendron is a genus of the magnolia family comprised of two flowering tree species that produce hardwoods of great ecological and economic value. However, only a limited amount of genetic research has been performed on the Liriodendron genus partly because transient or stable transgenic trees have been difficult to produce. In general, transient expression systems are indispensable for rapid, high-throughput screening and systematic characterization of gene functions at a low cost; therefore, development of such a system for Liriodendron would provide a necessary step forward for research on Magnoliaceae and other woody trees. Herein, we describe an efficient and rapid protocol for preparing protoplasts from the leaf mesophyll tissue of a Liriodendron hybrid and an optimized system for polyethylene glycol-mediated transient transfection of the protoplasts. Because the leaves of the Liriodendron hybrid are waxy, we formulated an enzyme mix containing 1.5% (w/v) Cellulase R-10, 0.5% (w/v) Macerozyme R-10, and 0.1% (w/v) Pectolyase Y-23 to efficiently isolate protoplasts from the Liriodendron hybrid leaf mesophyll tissue in 3 h. We optimized Liriodendron protoplast transfection efficiency by including 20 µg plasmid DNA per 104 protoplasts, a transformation time of 20 min, and inclusion of 20% (w/v) polyethylene glycol 4000. After integrating the Liriodendron WOX1 gene into pJIT166-GFP to produce a WOX1-GFP fusion product and transfecting it into isolated protoplasts, LhWOX1-GFP was found to localize to the nucleus according to its green fluorescence.
[Mh] Termos MeSH primário: Expressão Gênica
Liriodendron
Células do Mesofilo
Protoplastos
Transfecção
[Mh] Termos MeSH secundário: Técnicas de Cultura de Células
Vetores Genéticos
Proteínas Luminescentes/genética
Proteínas Luminescentes/metabolismo
Células do Mesofilo/metabolismo
Folhas de Planta
Proteínas de Plantas/genética
Proteínas de Plantas/metabolismo
Protoplastos/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Luminescent Proteins); 0 (Plant Proteins)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170823
[Lr] Data última revisão:
170823
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170322
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0172475


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[PMID]:28315633
[Au] Autor:Wiesmann V; Bergler M; Palmisano R; Prinzen M; Franz D; Wittenberg T
[Ad] Endereço:Fraunhofer Institute for Integrated Circuits IIS, Am Wolfsmantel 33, Erlangen, 91058, Germany. veit.wiesmann@iis.fraunhofer.de.
[Ti] Título:Using simulated fluorescence cell micrographs for the evaluation of cell image segmentation algorithms.
[So] Source:BMC Bioinformatics;18(1):176, 2017 Mar 18.
[Is] ISSN:1471-2105
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Manual assessment and evaluation of fluorescent micrograph cell experiments is time-consuming and tedious. Automated segmentation pipelines can ensure efficient and reproducible evaluation and analysis with constant high quality for all images of an experiment. Such cell segmentation approaches are usually validated and rated in comparison to manually annotated micrographs. Nevertheless, manual annotations are prone to errors and display inter- and intra-observer variability which influence the validation results of automated cell segmentation pipelines. RESULTS: We present a new approach to simulate fluorescent cell micrographs that provides an objective ground truth for the validation of cell segmentation methods. The cell simulation was evaluated twofold: (1) An expert observer study shows that the proposed approach generates realistic fluorescent cell micrograph simulations. (2) An automated segmentation pipeline on the simulated fluorescent cell micrographs reproduces segmentation performances of that pipeline on real fluorescent cell micrographs. CONCLUSION: The proposed simulation approach produces realistic fluorescent cell micrographs with corresponding ground truth. The simulated data is suited to evaluate image segmentation pipelines more efficiently and reproducibly than it is possible on manually annotated real micrographs.
[Mh] Termos MeSH primário: Algoritmos
Microscopia de Fluorescência
[Mh] Termos MeSH secundário: Animais
Linfócitos B/citologia
Linfócitos B/metabolismo
Forma Celular
Processamento de Imagem Assistida por Computador
Macrófagos/citologia
Macrófagos/metabolismo
Camundongos
Camundongos Endogâmicos C57BL
Protoplastos/citologia
Protoplastos/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170828
[Lr] Data última revisão:
170828
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170320
[St] Status:MEDLINE
[do] DOI:10.1186/s12859-017-1591-2


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[PMID]:28292294
[Au] Autor:Wu JZ; Liu Q; Geng XS; Li KM; Luo LJ; Liu JP
[Ad] Endereço:Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources, College of Agriculture, Hainan University, Haikou, Hainan Province, 570228, China.
[Ti] Título:Highly efficient mesophyll protoplast isolation and PEG-mediated transient gene expression for rapid and large-scale gene characterization in cassava (Manihot esculenta Crantz).
[So] Source:BMC Biotechnol;17(1):29, 2017 Mar 14.
[Is] ISSN:1472-6750
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Cassava (Manihot esculenta Crantz) is a major crop extensively cultivated in the tropics as both an important source of calories and a promising source for biofuel production. Although stable gene expression have been used for transgenic breeding and gene function study, a quick, easy and large-scale transformation platform has been in urgent need for gene functional characterization, especially after the cassava full genome was sequenced. METHODS: Fully expanded leaves from in vitro plantlets of Manihot esculenta were used to optimize the concentrations of cellulase R-10 and macerozyme R-10 for obtaining protoplasts with the highest yield and viability. Then, the optimum conditions (PEG4000 concentration and transfection time) were determined for cassava protoplast transient gene expression. In addition, the reliability of the established protocol was confirmed for subcellular protein localization. RESULTS: In this work we optimized the main influencing factors and developed an efficient mesophyll protoplast isolation and PEG-mediated transient gene expression in cassava. The suitable enzyme digestion system was established with the combination of 1.6% cellulase R-10 and 0.8% macerozyme R-10 for 16 h of digestion in the dark at 25 °C, resulting in the high yield (4.4 × 10 protoplasts/g FW) and vitality (92.6%) of mesophyll protoplasts. The maximum transfection efficiency (70.8%) was obtained with the incubation of the protoplasts/vector DNA mixture with 25% PEG4000 for 10 min. We validated the applicability of the system for studying the subcellular localization of MeSTP7 (an H /monosaccharide cotransporter) with our transient expression protocol and a heterologous Arabidopsis transient gene expression system. CONCLUSION: We optimized the main influencing factors and developed an efficient mesophyll protoplast isolation and transient gene expression in cassava, which will facilitate large-scale characterization of genes and pathways in cassava.
[Mh] Termos MeSH primário: Perfilação da Expressão Gênica/métodos
Melhoramento Genético/métodos
Manihot/genética
Proteínas de Plantas/genética
Engenharia de Proteínas/métodos
Protoplastos/citologia
[Mh] Termos MeSH secundário: Ensaios de Triagem em Larga Escala
Células do Mesofilo/citologia
Polietilenoglicóis
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Plant Proteins); 30IQX730WE (Polyethylene Glycols)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170613
[Lr] Data última revisão:
170613
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170316
[St] Status:MEDLINE
[do] DOI:10.1186/s12896-017-0349-2


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[PMID]:28284041
[Au] Autor:Stracke R; Turgut-Kara N; Weisshaar B
[Ad] Endereço:.
[Ti] Título:The AtMYB12 activation domain maps to a short C-terminal region of the transcription factor.
[So] Source:Z Naturforsch C;72(7-8):251-257, 2017 Jul 14.
[Is] ISSN:0939-5075
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:The Arabidopsis thaliana R2R3-MYB transcription factor MYB12 is a light-inducible, flavonol-specific activator of flavonoid biosynthesis. The transactivation activity of the AtMYB12 protein was analyzed using a C-terminal deletion series in a transient A. thaliana protoplast assay with the goal of mapping the activation domain (AD). Although the deletion of the last 46 C-terminal amino acids did not affect the activation capacity, the deletion of the last 98 amino acids almost totally abolished transactivation of two different target promoters. A domain swap experiment using the yeast GAL4 DNA-binding domain revealed that the region from positions 282 to 328 of AtMYB12 was sufficient for transactivation. In contrast to the R2R3-MYB ADs known thus far, that of AtMYB12 is not located at the rearmost C-terminal end of the protein. The AtMYB12 AD is conserved in other experimentally proven R2R3-MYB flavonol regulators from different species.
[Mh] Termos MeSH primário: Proteínas de Arabidopsis/genética
Arabidopsis/genética
Regulação da Expressão Gênica de Plantas
Fatores de Transcrição/genética
Ativação Transcricional
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Arabidopsis/metabolismo
Proteínas de Arabidopsis/metabolismo
Sítios de Ligação/genética
Flavonoides/biossíntese
Glucuronidase/genética
Glucuronidase/metabolismo
Mutação
Plantas Geneticamente Modificadas
Regiões Promotoras Genéticas/genética
Protoplastos/metabolismo
Plântulas/genética
Plântulas/metabolismo
Deleção de Sequência
Homologia de Sequência de Aminoácidos
Fatores de Transcrição/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Arabidopsis Proteins); 0 (Flavonoids); 0 (MYB12 protein, Arabidopsis); 0 (Transcription Factors); EC 3.2.1.31 (Glucuronidase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170927
[Lr] Data última revisão:
170927
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170312
[St] Status:MEDLINE


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[PMID]:28245335
[Au] Autor:Johnson GR; Kangas JD; Dovzhenko A; Trojok R; Voigt K; Majarian TD; Palme K; Murphy RF
[Ad] Endereço:Computational Biology Department, Carnegie Mellon University, Pittsburgh.
[Ti] Título:A method for characterizing phenotypic changes in highly variable cell populations and its application to high content screening of Arabidopsis thaliana protoplasts.
[So] Source:Cytometry A;91(4):326-335, 2017 Apr.
[Is] ISSN:1552-4930
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Quantitative image analysis procedures are necessary for the automated discovery of effects of drug treatment in large collections of fluorescent micrographs. When compared to their mammalian counterparts, the effects of drug conditions on protein localization in plant species are poorly understood and underexplored. To investigate this relationship, we generated a large collection of images of single plant cells after various drug treatments. For this, protoplasts were isolated from six transgenic lines of A. thaliana expressing fluorescently tagged proteins. Eight drugs at three concentrations were applied to protoplast cultures followed by automated image acquisition. For image analysis, we developed a cell segmentation protocol for detecting drug effects using a Hough transform-based region of interest detector and a novel cross-channel texture feature descriptor. In order to determine treatment effects, we summarized differences between treated and untreated experiments with an L Cramér-von Mises statistic. The distribution of these statistics across all pairs of treated and untreated replicates was compared to the variation within control replicates to determine the statistical significance of observed effects. Using this pipeline, we report the dose dependent drug effects in the first high-content Arabidopsis thaliana drug screen of its kind. These results can function as a baseline for comparison to other protein organization modeling approaches in plant cells. © 2017 International Society for Advancement of Cytometry.
[Mh] Termos MeSH primário: Arabidopsis
Processamento de Imagem Assistida por Computador/métodos
Protoplastos
[Mh] Termos MeSH secundário: Arabidopsis/efeitos dos fármacos
Fenótipo
Plantas Geneticamente Modificadas
Protoplastos/efeitos dos fármacos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170426
[Lr] Data última revisão:
170426
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170301
[St] Status:MEDLINE
[do] DOI:10.1002/cyto.a.23067



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