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We used video microscopy techniques as a tool for live examination of the dynamic aspects of plant/fungus interactions. Early, dynamic responses of epidermal midrib cells of leaves from a potato cultivar (Solanum tuberosum L. cv. Datura) carrying resistance gene R1 to Phytophthora infestans (race 1: compatible interaction, race 4: incompatible interaction) were monitored. Similar responses were observed in both types of interaction, ranging from no visible reaction of invaded plant cells to hypersensitive cell death. The overall defense response of each individual cell exhibited a highly dynamic behavior that appeared to be tightly coordinated with the growth of the fungus. Initial localized reactions, including major rearrangements within the cytoplasm, occurred directly at the fungal penetration site, where rapid apposition of autofluorescent material and callose took place. If fungal invasion stopped at this stage, the host cell restored its normal cytoplasmic activity and survived. Hypersensitive cell death occurred only when fungal growth had proceeded to the formation of a clearly identifiable haustorium. In such cases, cytoplasm and nucleus conglomerated around the intracellular fungal structure, followed by a sudden collapse of the whole conglomerate and an instantaneous collapse of the fungal haustorium. Only small quantitative differences between the compatible and incompatible interactions of the two fungal races were observed for these early responses of epidermal cells. In the incompatible interaction, a slightly larger number of epidermal cells responded to fungal attack. More pronounced quantitative differences between compatible and incompatible interactions occurred upon fungal invasion of the mesophyll. These differences in the number of responding cells were not reflected at the level of gene expression: the spatial and temporal activation patterns of two defense-related genes, encoding phenylalanine ammonia-lyase and pathogenesis-related protein 1, were similar in both types of interaction.Dedicated to Professor Peter Sitte, Freiburg, Germany, on the occasion of his 65th birthday 相似文献
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Michalis Barkoulas Jeroen S. van Zon Josselin Milloz Alexander van Oudenaarden Marie-Anne Félix 《Developmental cell》2013,24(1):64-75
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To avoid the specific problems concerning intrinsic membrane proteins in proteome analysis, an alternative strategy is described that is complementary to previous investigations using 2-D polyacrylamide gel electrophoresis (PAGE) techniques. The strategy involves (a) obtaining purified preparations of the membranes from Chlorobium tepidum by washing with 2 M NaBr, which removed membrane-associated soluble proteins and membrane-associated organelles; (b) separation of membrane protein complexes using 1-D Blue-native polyacrylamide gel electrophoresis (BN-PAGE) after solubilization with n-dodecyl-beta-d-maltoside (DDM); (c) combination of the BN with Tricine-SDS-PAGE; (d) high-throughput mass spectrometric analysis after gel band excision, in-gel digestion, and MALDI target spotting; and (e) protein identification from mixtures of tryptic peptides by peptide mass fingerprinting. Using this approach, we identified 143 different proteins, 70 of which have not been previously reported using 2-D PAGE techniques. Membrane proteins with up to 14 transmembrane helices were found, and this procedure proved to be efficient with proteins within a wide pI range (4.4-11.6). About 54% of the identified membrane proteins belong to various functional categories like energy metabolism, transport, signal transduction, and protein translocation, while for the others, a function is not yet known, indicating the potential of the method for the elucidation of the membrane proteomes in general. 相似文献
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Application of discriminant analysis and quantitative cytologic examination to gastric lesions 总被引:2,自引:0,他引:2
Karakitsos P Megalopoulou TM Pouliakis A Tzivras M Archimandritis A Kyroudes A 《Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology》2004,26(6):314-322
OBJECTIVE: To investigate of the potential value of morphometry and discriminant analysis for the classification of benign and malignant gastric cells and lesions. STUDY DESIGN: The data set consisted of 13,300 cells from 120 cases composed of 30 cases of cancer, 26 cases of gastritis and 64 cases of ulcer according to the final histologic diagnosis. The cytologic diagnosis was divided into 5 categories (gastritis, ulcer, inflammatory dysplasia, cancer and true dysplasia). Classification was attempted at 2 levels: the cell level to classify individual cells and the case level to classify individual cases. For the cellular classification the measured cells from 50% of available cases were selected as a training set to construct a model. The cells from the remaining cases were used as a test set to validate the model. Similarly for case classification, the same 50% of cases that were used for cell classification were used as a training set and the remaining cases as a test set. Images of routinely processed gastric smears stained by the Papanicolaou technique were analyzed by a customized image analysis system. RESULTS: Application of discriminant analysis on the test set gave correct classification of 98.4% of benign cells and 67.1% of malignant cells. On case classification, 100% accuracy was achieved for benign and malignant cases, both for the training and test sets. CONCLUSION: The application of discriminant analysis described in this paper could produce significant classification results at the cellular and individual case level. 相似文献
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Tsirogianni I Aivaliotis M Tsiotis G 《Zeitschrift für Naturforschung. C, Journal of biosciences》2004,59(1-2):132-134
The protein and lipid composition of a vitellin isolated from eggs of Sparus aurata were characterized by SDS PAGE, N-terminal sequence analysis and lipid analysis by thin layer chromatography and gas chromatography. The lipoprotein complex contains proteins with apparent molecular weights of 69, 59, 23, 21 and 12 kDa and were characterized as vitellinogenin fragments by N-terminal sequencing. Lipid extraction and analysis indicate an association of cholesterol and phospholipids with the protein subunits. The phospholipids contain fatty acids with 14, 16 and 18 carbon atoms as determined by GC/MS. 相似文献
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Georgios Hadjigeorgiou Efthimios Dardiotis Georgios Tsivgoulis Triantafyllos Doskas Damianos Petrou Nikolaos Makris Nikolaos Vlaikidis Thomas Thomaidis Athanasios Kyritsis Nikolaos Fakas Xoulietta Treska Clementine Karageorgiou Stefania Sotirli Christos Giannoulis Dimitra Papadimitriou Ioannis Mylonas Evaggelos Kouremenos George S. Vlachos Dimitrios Georgiopoulos Despoina Mademtzoglou Michalis Vikelis Elias Zintzaras 《PloS one》2015,10(10)