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Kazuhiro Kogame - One of the best experts on this subject based on the ideXlab platform.
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parthenogenesis is rare in the reproduction of a sexual field population of the isogamous brown alga Scytosiphon Scytosiphonaceae ectocarpales
Journal of Phycology, 2019Co-Authors: Masakazu Hoshino, Kazuhiro KogameAbstract:Parthenogenetic development of unfused gametes is commonly observed in laboratory cultures among various brown algal taxa. There is, however, little information on the contribution of parthenogenesis to the reproduction of field populations. In this study, we investigated whether parthenogenesis is present in a sexual population of the isogamous brown alga Scytosiphon with a 1:1 sex ratio. In culture, both female and male gametes showed higher mortality and slower development compared to zygotes. More than 90% of surviving partheno-germlings formed parthenosporophytes irrespective of the culture conditions tested. Therefore, if parthenogenesis occurs in the field, most unfused gametes are expected to form parthenosporophytes. Contrary to this expectation, parthenosporophytes were rare in the field population. We collected 126 sporophytic thalli and isolated and cultured a unilocular sporangium from each of them. We confirmed that cultures of 120 unilocular sporangia produced both female and male gametophytes by the observation of zygotes or amplification of PCR-based sex markers indicating that these sporangia originated from zygotic sporophytes. Only females were detected in cultures from two sporangia and only males from four sporangia suggesting that these sporangia originated from parthenosporophytes. In the Scytosiphon population, although parthenogenesis is observable in culture, our results demonstrate that the contribution of parthenogenesis to reproduction is small (≤4.8%) compared to sexual reproduction. Unfused gametes may not survive to form mature parthenosporophytes in significant numbers in the field partly due to their higher mortality and slower development compared from zygotes.
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Concordance between DNA-based species boundaries and reproductive isolating barriers in the Scytosiphon lomentaria species complex (Ectocarpales, Phaeophyceae)
Phycologia, 2018Co-Authors: Masakazu Hoshino, Shozo Ishikawa, Kazuhiro KogameAbstract:Abstract: DNA-based species delimitation methods often predict many putative cryptic species. Interpretation of these putative species (i.e. do they represent distinct species, or is this an erroneous estimate based on DNA variability?) is challenging because estimates of DNA-based delimitation are often difficult to verify using nongenetic data such as morphology or geographical distribution. In the present study, estimates of DNA-based delimitation methods were verified based on the biological species concept in Japanese populations of Scytosiphon lomentaria. Three DNA-based species delimitation methods (Generalised Mixed Yule Coalescent, Poisson Tree Processes and Automatic Barcode Gap Discovery) were conducted using mitochondrial (cox1) and nuclear (second intron of centrin gene) DNA sequence datasets. In the S. lomentaria species complex, five putative cryptic species (Ia–Va) were well-supported by DNA-based species delimitation, and these putative species were often found in the same locality. To ve...
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Genetic diversity and mitochondrial introgression in Scytosiphon lomentaria (Ectocarpales, Phaeophyceae) in the north-eastern Atlantic Ocean
Phycologia, 2015Co-Authors: Kazuhiro Kogame, Fabio Rindi, Akira F. Peters, Michael D. GuiryAbstract:Abstract: The common brown alga Scytosiphon lomentaria (Scytosiphonaceae) has generally been regarded as a species complex. In the Pacific, molecular data have shown that it includes more than one species; the diversity in other oceans, however, has not been examined. In the present study, the genetic diversity of S. lomentaria was investigated in 57 samples from the north-east Atlantic (mainly from Ireland) and two from the Mediterranean, which were compared with seven samples from Japan. Phylogenetic analyses based on the mitochondrial cytochrome oxidase subunit 3 gene (cox3) and nuclear ribosomal internal transcribed spacers (ITS1 and 2) revealed four separate monophyletic groups. Clades A1 and A2 consisted of the samples from the north-east Atlantic; the Mediterranean samples also belonged to A1. The affiliation of four samples was incongruent between cox3 and ITS trees, suggesting directional mitochondrial introgression from A2 to A1. Clades P1 and P2 contained nine samples from the north-east Atlant...
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disappearance of male mitochondrial dna after the four cell stage in sporophytes of the isogamous brown alga Scytosiphon lomentaria Scytosiphonaceae phaeophyceae 1
Journal of Phycology, 2010Co-Authors: Kei Kimura, Kazuhiro Kogame, Chikako Nagasato, Taizo MotomuraAbstract:Mitochondrial DNA (mtDNA) of the isogamous brown alga Scytosiphon lomentaria (Lyngb.) Link is inherited maternally. We used molecular biological and morphological analyses to investigate the fate of male mitochondria. Ultrastructural observations showed that the number of 25 mitochondria in a zygote coincided with the number of mitochondria derived from male and female gametes. This number remained almost constant during the first cell division. Strain-specific PCR in single germlings suggested that mtDNA derived from the female gamete remained in the germling during development, while the male mtDNA gradually and selectively disappeared after the four-cell stage. One week after fertilization, male mtDNA had disappeared in sporophytic cells. Using bisulfite DNA modification and methylation mapping assays, we found that the degree of methylation on three analyzed sites of mtDNA was not different between male and female gametes, suggesting that maternal inheritance of mtDNA is not defined by its methylation. This study indicates that the mechanism of selective elimination of male mtDNA is present in each cell of a four-celled sporophyte and that it does not depend on different degrees of DNA methylation between male and female mtDNA.
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Inheritance of mitochondrial and chloroplast genomes in the isogamous brown alga Scytosiphon lomentaria (Phaeophyceae)
Phycological Research, 2006Co-Authors: Yusuke Kato, Kazuhiro Kogame, Chikako Nagasato, Taizo MotomuraAbstract:SUMMARY Patterns of inheritance of chloroplasts and mitochondria were examined by fluorescence microscopy and haplotype genome markers in the isogamous brown alga Scytosiphon lomentaria (Lyngbye) Link. Germination of the zygote in this species was unilateral, the growing thallus developed entirely from the germ tube, and the original zygote cell did not develop except for the formation of a hair. Inheritance of chloroplasts was biparental, and partitioning of the two parental chloroplasts into the first sporophytic cells was accidental: either the maternal or the paternal chloroplast was migrated from the zygote into the germ tube cell, whereas the other chloroplast remained in the original cell. In contrast, the mitochondrial genome in all cells of the sporophyte came only from the female gamete (maternal inheritance). These inheritance patterns are similar to those of the isogamous brown alga Ectocarpus siliculosus (Dillwyn) Lyngbye. Maternal inheritance of mitochondria might be universal in brown algae.
Taizo Motomura - One of the best experts on this subject based on the ideXlab platform.
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disappearance of male mitochondrial dna after the four cell stage in sporophytes of the isogamous brown alga Scytosiphon lomentaria Scytosiphonaceae phaeophyceae 1
Journal of Phycology, 2010Co-Authors: Kei Kimura, Kazuhiro Kogame, Chikako Nagasato, Taizo MotomuraAbstract:Mitochondrial DNA (mtDNA) of the isogamous brown alga Scytosiphon lomentaria (Lyngb.) Link is inherited maternally. We used molecular biological and morphological analyses to investigate the fate of male mitochondria. Ultrastructural observations showed that the number of 25 mitochondria in a zygote coincided with the number of mitochondria derived from male and female gametes. This number remained almost constant during the first cell division. Strain-specific PCR in single germlings suggested that mtDNA derived from the female gamete remained in the germling during development, while the male mtDNA gradually and selectively disappeared after the four-cell stage. One week after fertilization, male mtDNA had disappeared in sporophytic cells. Using bisulfite DNA modification and methylation mapping assays, we found that the degree of methylation on three analyzed sites of mtDNA was not different between male and female gametes, suggesting that maternal inheritance of mtDNA is not defined by its methylation. This study indicates that the mechanism of selective elimination of male mtDNA is present in each cell of a four-celled sporophyte and that it does not depend on different degrees of DNA methylation between male and female mtDNA.
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Inheritance of mitochondrial and chloroplast genomes in the isogamous brown alga Scytosiphon lomentaria (Phaeophyceae)
Phycological Research, 2006Co-Authors: Yusuke Kato, Kazuhiro Kogame, Chikako Nagasato, Taizo MotomuraAbstract:SUMMARY Patterns of inheritance of chloroplasts and mitochondria were examined by fluorescence microscopy and haplotype genome markers in the isogamous brown alga Scytosiphon lomentaria (Lyngbye) Link. Germination of the zygote in this species was unilateral, the growing thallus developed entirely from the germ tube, and the original zygote cell did not develop except for the formation of a hair. Inheritance of chloroplasts was biparental, and partitioning of the two parental chloroplasts into the first sporophytic cells was accidental: either the maternal or the paternal chloroplast was migrated from the zygote into the germ tube cell, whereas the other chloroplast remained in the original cell. In contrast, the mitochondrial genome in all cells of the sporophyte came only from the female gamete (maternal inheritance). These inheritance patterns are similar to those of the isogamous brown alga Ectocarpus siliculosus (Dillwyn) Lyngbye. Maternal inheritance of mitochondria might be universal in brown algae.
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Identification and characterization of a fluorescent flagellar protein from the brown alga Scytosiphon lomentaria (Scytosiphonales, Phaeophyceae): A flavoprotein homologous to Old Yellow Enzyme
European Journal of Phycology, 2005Co-Authors: Satoshi Fujita, Taizo Motomura, Hiroshi Kawai, Shinya Yoshikawa, Mineo Iseki, Yumiko Makino, Masakatsu Watanabe, Akio MurakamiAbstract:The posterior flagellum of the zoospore of the brown alga Scytosiphon lomentaria exhibits bright green autofluorescence. To identify the fluorescent flagellar substance(s), we isolated flagella from zoospores and partially purified a flavoprotein by anion-exchange and gel-filtration chromatography. Spectrofluorometric and chromatographic analyses showed that the flavoprotein had an apparent molecular mass of 41 kDa and a non-covalently bound flavin mononucleotide as a chromophore. Based on partial amino acid sequences of the protein, a cDNA of the 41-kDa flavoprotein was cloned and sequenced. The deduced amino acid sequence of the cDNA was homologous to that of the Old Yellow Enzyme family distributed in proteobacteria, yeasts and vascular plants.
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Characterization of centrin genes from Ochromonas danica (Chrysophyceae) and Scytosiphon lomentaria (Phaeophyceae)
Phycological Research, 2004Co-Authors: Chikako Nagasato, Chihiro Uemori, Atsushi Kato, Taizo MotomuraAbstract:SUMMARY Centrin, the EF-hand Ca2+-binding protein is localized at the basal apparatus of flagella and in centrioles in many eukaryotic cells. In the present study, centrin genes of the heterokont algae have been clarified for the first time. We isolated and analyzed cDNA and genomic DNA of centrin genes from the crysophycean alga Ochromonas danica Prings (UTEX LB1298) and the brown alga Scytosiphon lomentaria (Lyngbye) Link. The centrin gene of Ochromonas contained an open reading frame of 163 amino acids. The deduced protein, named Odcen, exhibited 85%, 78% and 59% homology to Chlamydomonas, human and Arabidopsis centrin, respectively. The centrin genes of Scytosiphon contained an open reading frame of 164 amino acids. The deduced protein, named Slcen, exhibited 84%, 77% and 59% homology to Chlamydomonas, human and Arabidopsis centrin, respectively. Both Odcen and Slcen possessed N-terminal extensions before the conserved amino acid among various centrins, four EF-hand domains and an aromatic amino acid at the C-terminus. Southern blot hybridization suggested that the centrin gene occurs as a single copy gene in both Ochromonas and Scytosiphon genomes. Comparison of the sequence of the cDNA and the genomic DNA revealed that the Odcen gene was split into three fragments by introns and Slcen gene consisted of five fragments. The junctions of all introns of both genes conformed to the GT–AG rule. The introns of Slcen gene were considerably long and, as a result, the Slcen gene was approximately seven times longer than Odcen gene.
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Destruction of maternal centrioles during fertilization of the brown alga, Scytosiphon lomentaria (Scytosiphonales, Phaeophyceae)
Cell motility and the cytoskeleton, 2004Co-Authors: Chikako Nagasato, Taizo MotomuraAbstract:In brown algal fertilization, a pair of centrioles is derived from the male gamete, irrespective of the sexual reproduction pattern, i.e., isogamy, anisogamy, or oogamy. In this study, the manner in which the maternal centriole structure is destroyed in early zygotes of the isogamous brown alga Scytosiphon lomentaria was examined by electron microscopy. At fertilization, the zygote had two pairs of centrioles (flagellar basal bodies) derived from motile male and female gametes, and there was no morphological difference between the two pairs. The flagellar basal plate and the axonemal microtubules were still connected with the distal end of centrioles. Ultrastructural observations showed that the integrity of maternal-derived centrioles began to degenerate even in the 1-h-old zygote. At that time, the cylinder of triplet microtubules of the maternal centrioles became shorter from the distal end, and a section passing through the centrioles indicated that a part of the nine triplets of microtubules changed into doublet or singlet microtubules by degeneration of B and/or C tubules. In 2-h-old zygote, there was no trace of maternal centrioles ultrastructurally, and only the paternal centrioles remained. Further, reduction of centrin accompanying destruction of the maternal centrioles was examined in immunofluorescence microscopy. Centrin localized at the paternal and the maternal centrioles had the same fluorescence intensity in the early zygotes. At 4-6 h after fertilization, two spots indicating centrin localization showed different fluorescence intensity. Later, the weaker spot disappeared completely. These results showed that there is a difference in time between the destruction of the centriolar cylinders and the reduction of centrin molecules around them.
Chikako Nagasato - One of the best experts on this subject based on the ideXlab platform.
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disappearance of male mitochondrial dna after the four cell stage in sporophytes of the isogamous brown alga Scytosiphon lomentaria Scytosiphonaceae phaeophyceae 1
Journal of Phycology, 2010Co-Authors: Kei Kimura, Kazuhiro Kogame, Chikako Nagasato, Taizo MotomuraAbstract:Mitochondrial DNA (mtDNA) of the isogamous brown alga Scytosiphon lomentaria (Lyngb.) Link is inherited maternally. We used molecular biological and morphological analyses to investigate the fate of male mitochondria. Ultrastructural observations showed that the number of 25 mitochondria in a zygote coincided with the number of mitochondria derived from male and female gametes. This number remained almost constant during the first cell division. Strain-specific PCR in single germlings suggested that mtDNA derived from the female gamete remained in the germling during development, while the male mtDNA gradually and selectively disappeared after the four-cell stage. One week after fertilization, male mtDNA had disappeared in sporophytic cells. Using bisulfite DNA modification and methylation mapping assays, we found that the degree of methylation on three analyzed sites of mtDNA was not different between male and female gametes, suggesting that maternal inheritance of mtDNA is not defined by its methylation. This study indicates that the mechanism of selective elimination of male mtDNA is present in each cell of a four-celled sporophyte and that it does not depend on different degrees of DNA methylation between male and female mtDNA.
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Inheritance of mitochondrial and chloroplast genomes in the isogamous brown alga Scytosiphon lomentaria (Phaeophyceae)
Phycological Research, 2006Co-Authors: Yusuke Kato, Kazuhiro Kogame, Chikako Nagasato, Taizo MotomuraAbstract:SUMMARY Patterns of inheritance of chloroplasts and mitochondria were examined by fluorescence microscopy and haplotype genome markers in the isogamous brown alga Scytosiphon lomentaria (Lyngbye) Link. Germination of the zygote in this species was unilateral, the growing thallus developed entirely from the germ tube, and the original zygote cell did not develop except for the formation of a hair. Inheritance of chloroplasts was biparental, and partitioning of the two parental chloroplasts into the first sporophytic cells was accidental: either the maternal or the paternal chloroplast was migrated from the zygote into the germ tube cell, whereas the other chloroplast remained in the original cell. In contrast, the mitochondrial genome in all cells of the sporophyte came only from the female gamete (maternal inheritance). These inheritance patterns are similar to those of the isogamous brown alga Ectocarpus siliculosus (Dillwyn) Lyngbye. Maternal inheritance of mitochondria might be universal in brown algae.
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Characterization of centrin genes from Ochromonas danica (Chrysophyceae) and Scytosiphon lomentaria (Phaeophyceae)
Phycological Research, 2004Co-Authors: Chikako Nagasato, Chihiro Uemori, Atsushi Kato, Taizo MotomuraAbstract:SUMMARY Centrin, the EF-hand Ca2+-binding protein is localized at the basal apparatus of flagella and in centrioles in many eukaryotic cells. In the present study, centrin genes of the heterokont algae have been clarified for the first time. We isolated and analyzed cDNA and genomic DNA of centrin genes from the crysophycean alga Ochromonas danica Prings (UTEX LB1298) and the brown alga Scytosiphon lomentaria (Lyngbye) Link. The centrin gene of Ochromonas contained an open reading frame of 163 amino acids. The deduced protein, named Odcen, exhibited 85%, 78% and 59% homology to Chlamydomonas, human and Arabidopsis centrin, respectively. The centrin genes of Scytosiphon contained an open reading frame of 164 amino acids. The deduced protein, named Slcen, exhibited 84%, 77% and 59% homology to Chlamydomonas, human and Arabidopsis centrin, respectively. Both Odcen and Slcen possessed N-terminal extensions before the conserved amino acid among various centrins, four EF-hand domains and an aromatic amino acid at the C-terminus. Southern blot hybridization suggested that the centrin gene occurs as a single copy gene in both Ochromonas and Scytosiphon genomes. Comparison of the sequence of the cDNA and the genomic DNA revealed that the Odcen gene was split into three fragments by introns and Slcen gene consisted of five fragments. The junctions of all introns of both genes conformed to the GT–AG rule. The introns of Slcen gene were considerably long and, as a result, the Slcen gene was approximately seven times longer than Odcen gene.
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Destruction of maternal centrioles during fertilization of the brown alga, Scytosiphon lomentaria (Scytosiphonales, Phaeophyceae)
Cell motility and the cytoskeleton, 2004Co-Authors: Chikako Nagasato, Taizo MotomuraAbstract:In brown algal fertilization, a pair of centrioles is derived from the male gamete, irrespective of the sexual reproduction pattern, i.e., isogamy, anisogamy, or oogamy. In this study, the manner in which the maternal centriole structure is destroyed in early zygotes of the isogamous brown alga Scytosiphon lomentaria was examined by electron microscopy. At fertilization, the zygote had two pairs of centrioles (flagellar basal bodies) derived from motile male and female gametes, and there was no morphological difference between the two pairs. The flagellar basal plate and the axonemal microtubules were still connected with the distal end of centrioles. Ultrastructural observations showed that the integrity of maternal-derived centrioles began to degenerate even in the 1-h-old zygote. At that time, the cylinder of triplet microtubules of the maternal centrioles became shorter from the distal end, and a section passing through the centrioles indicated that a part of the nine triplets of microtubules changed into doublet or singlet microtubules by degeneration of B and/or C tubules. In 2-h-old zygote, there was no trace of maternal centrioles ultrastructurally, and only the paternal centrioles remained. Further, reduction of centrin accompanying destruction of the maternal centrioles was examined in immunofluorescence microscopy. Centrin localized at the paternal and the maternal centrioles had the same fluorescence intensity in the early zygotes. At 4-6 h after fertilization, two spots indicating centrin localization showed different fluorescence intensity. Later, the weaker spot disappeared completely. These results showed that there is a difference in time between the destruction of the centriolar cylinders and the reduction of centrin molecules around them.
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NEW PYRENOID FORMATION IN THE BROWN ALGA, Scytosiphon LOMENTARIA (ScytosiphonALES, PHAEOPHYCEAE)1
Journal of Phycology, 2002Co-Authors: Chikako Nagasato, Taizo MotomuraAbstract:The Scytosiphon lomentaria (Lyngbye) Link cell characteristically has only one chloroplast with a prominent protruding pyrenoid. We observed the appearance of a new pyrenoid in each chloroplast during first mitosis in zygotes of S. lomentaria, using the freeze substitution technique. At first, a pyrenoid matrix appeared within the outermost stroma, in which thylakoid triplets and ribosomes were absent. At this time, the surface of this part remained smooth. The old pyrenoid was covered with a pyrenoid cap on the cytoplasmic side, whereas there was no pyrenoid cap on the new pyrenoid before protrusion. Irregularly shaped membranous sacs containing fine granular materials associated with the cytoplasmic side of the new pyrenoid. The sacs fused with each other and changed conformation and finally transformed into the pyrenoid cap. The new pyrenoid gradually protruded toward the cytoplasm, and the new pyrenoid cap became curved along the surface of pyrenoid. Cytokinesis occurred, and each chloroplast had two prominent protruding pyrenoids in two-celled zygotes. We examined immunolocalization of β-1,3-glucans within the pyrenoid cap with a monoclonal antibody, using EM. Gold particles indicating localization of β-1,3-glucans were detected in vacuoles but never in the pyrenoid cap. This observation suggests that the pyrenoid cap in brown algae contains no photosynthetic products such as polysaccharide.
Juan A. Correa - One of the best experts on this subject based on the ideXlab platform.
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Seaweeds early development: detrimental effects of desiccation and attenuation by algal extracts
Planta, 2011Co-Authors: Loretto Contreras-porcia, Daniela Thomas, Sebastián Callejas, Aníbal Contreras, Ana Lafuente, María Rosa Flores-molina, Camille Sordet, Georg Pohnert, Juan A. CorreaAbstract:The effects of desiccation on the early development stages of Mazzaella laminarioides, Scytosiphon lomentaria and Lessonia nigrescens, algal species with different patterns of distribution across the intertidal zone, were examined in the laboratory. In addition, the protective effect against desiccation was evaluated using algal extracts, including those from Porphyra columbina, a macroalga tolerant to desiccation that lives in the uppermost part of the intertidal zone. Our results showed that M. laminarioides displayed the highest resistance to daily desiccation, followed by S. lomentaria, whereas L. nigrescens was the most susceptible. Spores from L. nigrescens exposed to desiccation, although being able to germinate, ceased further post-germination development. In addition, our results showed that all species exposed to extracts from desiccated P. columbina successfully completed their development and strongly suggest the occurrence of compounds with protective properties that help in attenuating the stress caused by desiccation. Finally, our results indicate that the magnitude of the effects generated by desiccation on the early algal development is related to the position of the species in the intertidal zone, and that the protective effects of P. columbina extracts reveal an exceptional metabolism of this species under desiccation stress.
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proteomic analysis and identification of copper stress regulated proteins in the marine alga Scytosiphon gracilis phaeophyceae
Aquatic Toxicology, 2010Co-Authors: Loretto Contreras, Alejandra Moenne, Philippe Potin, Fanny Gaillard, Juan A. CorreaAbstract:Abstract A proteomic analysis combining peptide de novo sequencing and BLAST analysis was used to identify novel proteins involved in copper tolerance in the marine alga Scytosiphon gracilis (Phaeophyceae). Algal material was cultivated in seawater without copper (control) or supplemented with 100 μg L−1 for 4 days, and protein extracts were separated by two-dimensional gel electrophoresis (2-DE). From the proteins obtained in the copper treatment, 25 over-expressed, 5 under-expressed and 5 proteins with no changes as compared with the control, were selected for sequencing. Tryptic-peptides obtained from 35 spots were analyzed by capillary liquid chromatography and tandem mass spectroscopy (capLC/MS/MS), and protein identity was determined by BLASTP. We identified 19 over-expressed proteins, including a chloroplast peroxiredoxin, a cytosolic phosphomannomutase, a cytosolic glyceraldehyde-3-phosphate dehydrogenase, 3 ABC transporters, a chaperonine, a subunit of the proteasome and a tRNA synthase, among others. The possible involvement of these over-expressed proteins in buffering oxidative stress and avoiding metal uptake in S. gracilis exposed to copper excess is discussed taking into consideration the information available for other plant models.
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Two-dimensional gel electrophoresis analysis of brown algal protein extracts
Journal of Phycology, 2008Co-Authors: Andrés Ritter, Juan A. Correa, Alejandra Moenne, Geraldine Dennett, Freddy Boehmwald, Nathalie Guitton, Charles Pineau, Philippe Potin, Loretto ContrerasAbstract:High-quality protein extracts are required for proteomic studies, a field that is poorly developed for marine macroalgae. A reliable phenol extraction protocol using Scytosiphon gracilis Kogame and Ectocarpus siliculosus (Dillwyn) Lyngb. (Phaeophyceae) as algal models resulted in high-quality protein extracts. The performance of the new protocol was tested against four methods available for vascular plants and a seaweed. The protocol, which includes an initial step to remove salts from the algal tissues, allowed the use of highly resolving two-dimensional gel electrophoresis (2-DE) protein analyses, providing the opportunity to unravel potentially novel physiological processes unique to this group of marine organisms.
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Molecular and morphologically distinct Scytosiphon species (Scytosiphonales, Phaeophyceae) display similar antioxidant capacities
Journal of Phycology, 2007Co-Authors: Loretto Contreras, Alejandra Moenne, Geraldine Dennett, R. Eduardo Palma, Juan A. CorreaAbstract:The taxonomic status of an alga with complanate thalli, occurring in central Chile and belonging to the genus Scytosiphon, was elucidated. Morphological and molecular features demonstrated that, in addition to the known and widespread constricted S. lomentaria (Lyngb.) Link that occurs along the Chilean coast, there is a Scytosiphon with complanate thalli that occurs only in central Chile—S. gracilis Kogame. Morphological analyses of this previously unreported complanate Scytosiphon showed thalli without constrictions, coherent plurilocular sporangia without ascocysts, and phaeophycean hairs arising from cortical cells. Furthermore, sequences of the internal transcribed spacer (ITS1 and ITS2) regions of the complanate Scytosiphon were 99.8% and 100% identical to those detected in S. gracilis from Korea. ITS-based comparative analyses showed that complanate Scytosiphon grouped in a different clade than S. lomentaria and S. tenellus Kogame from various parts of the world, including Chilean species. Moreover, molecular analyses suggest the occurrence of two distinct ITS types of S. lomentaria in northern Chile, corresponding to the Korean and Greek types. On the other hand, biochemical analyses of copper-induced antioxidant responses in S. gracilis and S. lomentaria showed an identical increase in antioxidant enzyme activities. These results suggest that copper tolerance might be a constitutive trait in these species of Scytosiphon.
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antioxidant responses in Scytosiphon lomentaria phaeophyceae inhabiting copper enriched coastal environments
Journal of Phycology, 2005Co-Authors: Loretto Contreras, Alejandra Moenne, Juan A. CorreaAbstract:Scytosiphon lomentaria (Lingb.) Link. (Phaeophyceae) is one of the two dominant seaweeds in a coastal area of northern Chile affected by copper mine wastes, where the concentration of copper in water and algal tissues remains higher than in non-impacted sites. Copper-loaded plants develop oxidative stress, as demonstrated by the increased levels of reactive oxygen species and lipoperoxides. This stress was associated with 1) an enhanced activity of the antioxidant enzymes catalase, glutathione peroxidase, ascorbate peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase and 2) an inhibition of the glutathione reductase activity. Furthermore, stressed plants showed a decrease in glutathione and phenolic compounds levels and an increase in total ascorbate. Reciprocal transplants revealed that plants rapidly adjusted their antioxidant system in response to the conditions of the receiving site. In individuals transplanted from the copper-enriched environment to the control site, normal levels of lipoperoxides and antioxidant compounds were restored in 48 h and antioxidant enzymes recovered their basal activities in 96 h. Individuals transplanted from the control site to the copper-enriched area adjusted their antioxidant compounds and antioxidant enzymes within 48 h and 96 h, respectively, and reached the functional status of the local plants. We conclude that S. lomentaria inhabiting the copper-enriched area buffered oxidative stress by a simultaneous involvement of antioxidant enzymes and water-soluble antioxidant compounds. These antioxidant responses were rapid and reversible, suggesting that copper resistance in S. lomentaria is a constitutive trait and that copper enrichment of the area did not result in a locally adapted copper-tolerant ecotype.
Loretto Contreras - One of the best experts on this subject based on the ideXlab platform.
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proteomic analysis and identification of copper stress regulated proteins in the marine alga Scytosiphon gracilis phaeophyceae
Aquatic Toxicology, 2010Co-Authors: Loretto Contreras, Alejandra Moenne, Philippe Potin, Fanny Gaillard, Juan A. CorreaAbstract:Abstract A proteomic analysis combining peptide de novo sequencing and BLAST analysis was used to identify novel proteins involved in copper tolerance in the marine alga Scytosiphon gracilis (Phaeophyceae). Algal material was cultivated in seawater without copper (control) or supplemented with 100 μg L−1 for 4 days, and protein extracts were separated by two-dimensional gel electrophoresis (2-DE). From the proteins obtained in the copper treatment, 25 over-expressed, 5 under-expressed and 5 proteins with no changes as compared with the control, were selected for sequencing. Tryptic-peptides obtained from 35 spots were analyzed by capillary liquid chromatography and tandem mass spectroscopy (capLC/MS/MS), and protein identity was determined by BLASTP. We identified 19 over-expressed proteins, including a chloroplast peroxiredoxin, a cytosolic phosphomannomutase, a cytosolic glyceraldehyde-3-phosphate dehydrogenase, 3 ABC transporters, a chaperonine, a subunit of the proteasome and a tRNA synthase, among others. The possible involvement of these over-expressed proteins in buffering oxidative stress and avoiding metal uptake in S. gracilis exposed to copper excess is discussed taking into consideration the information available for other plant models.
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Two-dimensional gel electrophoresis analysis of brown algal protein extracts
Journal of Phycology, 2008Co-Authors: Andrés Ritter, Juan A. Correa, Alejandra Moenne, Geraldine Dennett, Freddy Boehmwald, Nathalie Guitton, Charles Pineau, Philippe Potin, Loretto ContrerasAbstract:High-quality protein extracts are required for proteomic studies, a field that is poorly developed for marine macroalgae. A reliable phenol extraction protocol using Scytosiphon gracilis Kogame and Ectocarpus siliculosus (Dillwyn) Lyngb. (Phaeophyceae) as algal models resulted in high-quality protein extracts. The performance of the new protocol was tested against four methods available for vascular plants and a seaweed. The protocol, which includes an initial step to remove salts from the algal tissues, allowed the use of highly resolving two-dimensional gel electrophoresis (2-DE) protein analyses, providing the opportunity to unravel potentially novel physiological processes unique to this group of marine organisms.
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Molecular and morphologically distinct Scytosiphon species (Scytosiphonales, Phaeophyceae) display similar antioxidant capacities
Journal of Phycology, 2007Co-Authors: Loretto Contreras, Alejandra Moenne, Geraldine Dennett, R. Eduardo Palma, Juan A. CorreaAbstract:The taxonomic status of an alga with complanate thalli, occurring in central Chile and belonging to the genus Scytosiphon, was elucidated. Morphological and molecular features demonstrated that, in addition to the known and widespread constricted S. lomentaria (Lyngb.) Link that occurs along the Chilean coast, there is a Scytosiphon with complanate thalli that occurs only in central Chile—S. gracilis Kogame. Morphological analyses of this previously unreported complanate Scytosiphon showed thalli without constrictions, coherent plurilocular sporangia without ascocysts, and phaeophycean hairs arising from cortical cells. Furthermore, sequences of the internal transcribed spacer (ITS1 and ITS2) regions of the complanate Scytosiphon were 99.8% and 100% identical to those detected in S. gracilis from Korea. ITS-based comparative analyses showed that complanate Scytosiphon grouped in a different clade than S. lomentaria and S. tenellus Kogame from various parts of the world, including Chilean species. Moreover, molecular analyses suggest the occurrence of two distinct ITS types of S. lomentaria in northern Chile, corresponding to the Korean and Greek types. On the other hand, biochemical analyses of copper-induced antioxidant responses in S. gracilis and S. lomentaria showed an identical increase in antioxidant enzyme activities. These results suggest that copper tolerance might be a constitutive trait in these species of Scytosiphon.
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antioxidant responses in Scytosiphon lomentaria phaeophyceae inhabiting copper enriched coastal environments
Journal of Phycology, 2005Co-Authors: Loretto Contreras, Alejandra Moenne, Juan A. CorreaAbstract:Scytosiphon lomentaria (Lingb.) Link. (Phaeophyceae) is one of the two dominant seaweeds in a coastal area of northern Chile affected by copper mine wastes, where the concentration of copper in water and algal tissues remains higher than in non-impacted sites. Copper-loaded plants develop oxidative stress, as demonstrated by the increased levels of reactive oxygen species and lipoperoxides. This stress was associated with 1) an enhanced activity of the antioxidant enzymes catalase, glutathione peroxidase, ascorbate peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase and 2) an inhibition of the glutathione reductase activity. Furthermore, stressed plants showed a decrease in glutathione and phenolic compounds levels and an increase in total ascorbate. Reciprocal transplants revealed that plants rapidly adjusted their antioxidant system in response to the conditions of the receiving site. In individuals transplanted from the copper-enriched environment to the control site, normal levels of lipoperoxides and antioxidant compounds were restored in 48 h and antioxidant enzymes recovered their basal activities in 96 h. Individuals transplanted from the control site to the copper-enriched area adjusted their antioxidant compounds and antioxidant enzymes within 48 h and 96 h, respectively, and reached the functional status of the local plants. We conclude that S. lomentaria inhabiting the copper-enriched area buffered oxidative stress by a simultaneous involvement of antioxidant enzymes and water-soluble antioxidant compounds. These antioxidant responses were rapid and reversible, suggesting that copper resistance in S. lomentaria is a constitutive trait and that copper enrichment of the area did not result in a locally adapted copper-tolerant ecotype.
