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Juan Francisco Jiménez-bremont - One of the best experts on this subject based on the ideXlab platform.
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PEST sequences from a cactus dehydrin regulate its proteolytic degradation
PeerJ Inc., 2019Co-Authors: Adriana L. Salazar-retana, Israel Maruri-lópez, Itzell E. Hernández-sánchez, Alicia Becerra-flora, María De La Luz Guerrero-gonzález, Juan Francisco Jiménez-bremontAbstract:Dehydrins (DHNs) are intrinsically disordered proteins expressed under cellular dehydration-related stresses. In this study, we identified potential proteolytic PEST sequences located at the central and C-terminal regions from the Opuntia streptacantha OpsDHN1 protein. In order to evaluate these PEST sequences as proteolytic tags, we generated a translational fusion with the GUS reporter protein and OpsDHN1 coding sequence. We found a GUS degradation effect in tobacco agro-infiltrated leaves and Arabidopsis transgenic lines that expressed the fusion GUS::OpsDHN1 full-length. Also, two additional translational fusions between OpsDHN1 protein fragments that include the central (GUS::PEST-1) or the C-terminal (GUS::PEST-2) PEST sequences were able to decrease the GUS activity, with PEST-2 showing the greatest reduction in GUS activity. GUS signal was abated when the OpsDHN1 fragment that includes both PEST sequences (GUS::PEST-1-2) were fused to GUS. Treatment with the MG132 proteasome inhibitor attenuated the PEST-mediated GUS degradation. Point mutations of phosphorylatable residues in PEST sequences reestablished GUS signal, hence these sequences are important during protein degradation. Finally, in silico analysis identified potential PEST sequences in other plant DHNs. This is the first study reporting presence of PEST motifs in dehydrins
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Continuous high and low temperature induced a decrease of photosynthetic activity and changes in the diurnal fluctuations of organic acids in Opuntia streptacantha
PloS one, 2017Co-Authors: Zaida Zarely Ojeda-pérez, Juan Francisco Jiménez-bremont, Pablo Delgado-sánchezAbstract:Opuntia plants grow naturally in areas where temperatures are extreme and highly variable in the day during the entire year. These plants survive through different adaptations to respond to adverse environmental conditions. Despite this capability, it is unknown how CAM photosynthetic activity and growth in Opuntia plantlets is affected by constant heat or cold. Therefore, the main objective of this research was to evaluate the short-term effect of high (40°C) and low (4°C) continuous temperatures on the photosynthetic efficiency, the organic acid content (malic acid) and the relative growth rate (RGR) in seven-month-old Opuntia streptacantha plantlets during 5, 10, and 15 days. Chlorophyll fluorescence analysis allowed us to determine that high temperatures negatively impact the photosynthetic efficiency of O. streptacantha plantlets, which exhibited the lowest values of maximum quantum efficiency of the photosystem II (Fv/Fm = 52%, Fv/F0 = 85%), operational quantum yield of PS (ΦPSII = 65%) and relative electron transport rate (rETR = 65%), as well as highest values of basal fluorescence (F0 = 226%) during 15 days of treatment. Similarly, low temperatures decreased Fv/Fm (16%), Fv/F0 (50%), ΦPSII and rETR (16%). High temperatures also decreased nocturnal acidification in approximately 34-50%, whereas low temperatures increased it by 30-36%. Additionally, both continuous temperatures affected drastically diurnal consumption of malic acid, which was related to a significant RGR inhibition, where the specific photosynthetic structure area component was the most affected. Our results allowed determining that, despite the high tolerance to extreme temperatures described for Opuntia plants, young individuals of O. streptacantha suffered photosynthetic impairment that led to the inhibition of their growth. Thus, the main findings reported in this study can help to predict the potential impact of climatic change on the establishment and survival of succulent species of arid and semiarid regions of Mexico.
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Effects of catalase on chloroplast arrangement in Opuntia streptacantha chlorenchyma cells under salt stress.
Scientific reports, 2017Co-Authors: Diana Marcela Arias-moreno, Juan Francisco Jiménez-bremont, Israel Maruri-lópez, Pablo Delgado-sánchezAbstract:In arid and semiarid regions, low precipitation rates lead to soil salinity problems, which may limit plant establishment, growth, and survival. Herein, we investigated the NaCl stress effect on chlorophyll fluorescence, photosynthetic-pigments, movement and chloroplasts ultrastructure in chlorenchyma cells of Opuntia streptacantha cladodes. Cladodes segments were exposed to salt stress at 0, 100, 200, and 300 mM NaCl for 8, 16, and 24 h. The results showed that salt stress reduced chlorophyll content, F v /F m , ΦPSII, and qP values. Under the highest salt stress treatments, the chloroplasts were densely clumped toward the cell center and thylakoid membranes were notably affected. We analyzed the effect of exogenous catalase in salt-stressed cladode segments during 8, 16, and 24 h. The catalase application to salt-stressed cladodes counteracted the NaCl adverse effects, increasing the chlorophyll fluorescence parameters, photosynthetic-pigments, and avoided chloroplast clustering. Our results indicate that salt stress triggered the chloroplast clumping and affected the photosynthesis in O. streptacantha chlorenchyma cells. The exogenous catalase reverted the H2O2 accumulation and clustering of chloroplast, which led to an improvement of the photosynthetic efficiency. These data suggest that H2O2 detoxification by catalase is important to protect the chloroplast, thus conserving the photosynthetic activity in O. streptacantha under stress.
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Effect of extreme temperature treatments on RGR and its components of Opuntia streptacantha plantlets of seven-month-old.
2017Co-Authors: Zaida Zarely Ojeda-pérez, Juan Francisco Jiménez-bremont, Pablo Delgado-sánchezAbstract:Effect of extreme temperature treatments on RGR and its components of Opuntia streptacantha plantlets of seven-month-old.
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A dehydrin-dehydrin interaction: the case of SK3 from Opuntia streptacantha
Frontiers in plant science, 2014Co-Authors: Itzell E. Hernández-sánchez, Steffen P. Graether, David M. Martynowicz, Aída Araceli Rodríguez-hernández, Maria B. Pérez-morales, Juan Francisco Jiménez-bremontAbstract:Dehydrins belongs to a large group of highly hydrophilic proteins known as Late Embryogenesis Abundant (LEA) proteins. It is well know that dehydrins are intrinsically disordered plant proteins that accumulate during the late stages of embryogenesis and in response to abiotic stresses; however, the molecular mechanisms by which their functions are carried out are still unclear. We have previously reported that transgenic Arabidopsis plants overexpressing an Opuntia streptacantha SK3 dehydrin (OpsDHN1) show enhanced tolerance to freezing stress. Herein, we show using a split-ubiquitin yeast two-hybrid system that OpsDHN1 dimerizes. We found that the deletion of regions containing K-segments and the histidine-rich region in the OpsDHN1 protein affects dimer formation. Not surprisingly, in silico protein sequence analysis suggests that OpsDHN1 is an intrinsically disordered protein, an observation that was confirmed by circular dichroism and gel filtration of the recombinantly expressed protein. The addition of zinc triggered the association of recombinantly expressed OpsDHN1 protein, likely through its histidine-rich motif. These data brings new insights about the molecular mechanism of the OpsDHN1 SK3-dehydrin
Pablo Delgado-sánchez - One of the best experts on this subject based on the ideXlab platform.
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Continuous high and low temperature induced a decrease of photosynthetic activity and changes in the diurnal fluctuations of organic acids in Opuntia streptacantha
PloS one, 2017Co-Authors: Zaida Zarely Ojeda-pérez, Juan Francisco Jiménez-bremont, Pablo Delgado-sánchezAbstract:Opuntia plants grow naturally in areas where temperatures are extreme and highly variable in the day during the entire year. These plants survive through different adaptations to respond to adverse environmental conditions. Despite this capability, it is unknown how CAM photosynthetic activity and growth in Opuntia plantlets is affected by constant heat or cold. Therefore, the main objective of this research was to evaluate the short-term effect of high (40°C) and low (4°C) continuous temperatures on the photosynthetic efficiency, the organic acid content (malic acid) and the relative growth rate (RGR) in seven-month-old Opuntia streptacantha plantlets during 5, 10, and 15 days. Chlorophyll fluorescence analysis allowed us to determine that high temperatures negatively impact the photosynthetic efficiency of O. streptacantha plantlets, which exhibited the lowest values of maximum quantum efficiency of the photosystem II (Fv/Fm = 52%, Fv/F0 = 85%), operational quantum yield of PS (ΦPSII = 65%) and relative electron transport rate (rETR = 65%), as well as highest values of basal fluorescence (F0 = 226%) during 15 days of treatment. Similarly, low temperatures decreased Fv/Fm (16%), Fv/F0 (50%), ΦPSII and rETR (16%). High temperatures also decreased nocturnal acidification in approximately 34-50%, whereas low temperatures increased it by 30-36%. Additionally, both continuous temperatures affected drastically diurnal consumption of malic acid, which was related to a significant RGR inhibition, where the specific photosynthetic structure area component was the most affected. Our results allowed determining that, despite the high tolerance to extreme temperatures described for Opuntia plants, young individuals of O. streptacantha suffered photosynthetic impairment that led to the inhibition of their growth. Thus, the main findings reported in this study can help to predict the potential impact of climatic change on the establishment and survival of succulent species of arid and semiarid regions of Mexico.
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Effects of catalase on chloroplast arrangement in Opuntia streptacantha chlorenchyma cells under salt stress.
Scientific reports, 2017Co-Authors: Diana Marcela Arias-moreno, Juan Francisco Jiménez-bremont, Israel Maruri-lópez, Pablo Delgado-sánchezAbstract:In arid and semiarid regions, low precipitation rates lead to soil salinity problems, which may limit plant establishment, growth, and survival. Herein, we investigated the NaCl stress effect on chlorophyll fluorescence, photosynthetic-pigments, movement and chloroplasts ultrastructure in chlorenchyma cells of Opuntia streptacantha cladodes. Cladodes segments were exposed to salt stress at 0, 100, 200, and 300 mM NaCl for 8, 16, and 24 h. The results showed that salt stress reduced chlorophyll content, F v /F m , ΦPSII, and qP values. Under the highest salt stress treatments, the chloroplasts were densely clumped toward the cell center and thylakoid membranes were notably affected. We analyzed the effect of exogenous catalase in salt-stressed cladode segments during 8, 16, and 24 h. The catalase application to salt-stressed cladodes counteracted the NaCl adverse effects, increasing the chlorophyll fluorescence parameters, photosynthetic-pigments, and avoided chloroplast clustering. Our results indicate that salt stress triggered the chloroplast clumping and affected the photosynthesis in O. streptacantha chlorenchyma cells. The exogenous catalase reverted the H2O2 accumulation and clustering of chloroplast, which led to an improvement of the photosynthetic efficiency. These data suggest that H2O2 detoxification by catalase is important to protect the chloroplast, thus conserving the photosynthetic activity in O. streptacantha under stress.
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Effect of extreme temperature treatments on RGR and its components of Opuntia streptacantha plantlets of seven-month-old.
2017Co-Authors: Zaida Zarely Ojeda-pérez, Juan Francisco Jiménez-bremont, Pablo Delgado-sánchezAbstract:Effect of extreme temperature treatments on RGR and its components of Opuntia streptacantha plantlets of seven-month-old.
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Ecophysiological and Anatomical Mechanisms behind the Nurse Effect: Which Are More Important? A Multivariate Approach for Cactus Seedlings
PloS one, 2013Co-Authors: Pablo Delgado-sánchez, Juan Francisco Jiménez-bremont, Leonardo Chapa-vargas, Laura Yáñez-espinosa, Joel FloresAbstract:Background Cacti establish mostly occurs under the canopy of nurse plants which provide a less stressful micro-environment, although mechanisms underlying this process are unknown. The impact of the combination of light and watering treatments on Opuntia streptacantha (Cactaceae) seedlings was examined. Methods/Principal Findings Ecophysiological [titratable acidity, osmotic potential (‘solute potential’, Ψs), relative growth rate (RGR) and their components (NAR, SLA, and LWR)], anatomical (chloroplast density, chloroplast frequency, and cell area), and environmental [photosynthetic photon flux density (PPFD) and air temperature] sets of variables were analyzed, assessing relationships between them and measuring the intensity of the relationships. Three harvests were carried out at days 15, 30, and 45. Ψs and acidity content were the most important responses for seedling establishment. The main anatomical and environmental variables were chloroplast density and water availability, respectively. Opuntia streptacantha seedlings establish better in the shade-watering treatment, due to higher Ψs and acidity, unaffected chloroplasts, and lower PPFD. In addition, the chloroplasts of cells under high-light and non-watering treatment were clumped closer to the center of the cytosol than those under shade-drought, to avoid photoinhibition and/or to better distribute or utilize the penetrating light in the green plant tissue. Conclusions Opuntia seedlings grow better under the shade, although they can tolerate drought in open spaces by increasing and moving chloroplasts and avoiding drastic decreases in their Ψs. This tolerance could have important implications for predicting the impact of climate change on natural desert regeneration, as well as for planning reforestation-afforestation practices, and rural land uses.
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LEA Gene Introns: is the Intron of Dehydrin Genes a Characteristic of the Serine-Segment?
Plant Molecular Biology Reporter, 2013Co-Authors: Juan Francisco Jiménez-bremont, Israel Maruri-lópez, Ana Erika Ochoa-alfaro, Pablo Delgado-sánchez, Jaime Bravo, Margarita Rodríguez-kesslerAbstract:Dehydrins, which belong to group 2 LEA proteins, are a family of intrinsically unstructured plant proteins that accumulate during the late stages of embryogenesis and in response to abiotic stresses. We have previously reported that the OpsDHN1 gene, encoding an SK_3-type acidic dehydrin protein from Opuntia streptacantha , contains an intron inserted within the sequence encoding the S -motif. Herein, we present an in silico analysis of intron sequences in dehydrin genes from mono- and dicotyledonous plants that reveals a preference for insertion within the nucleotide sequence encoding the S -motif. Sequence comparison of ten Dhn genes from Arabidopsis thaliana and the orthologous genes in Arabidopsis lyrata revealed that introns maintain considerable sequence identity and conserve the insertion pattern. Furthermore, syntenic regions were identified among eight orthologous genes of A. thaliana and A. lyrata , showing that correlated gene arrangements are conserved between these Arabidopsis species. Our study shows that most SKn-type dehydrins contain one intron that is conserved in phase and location; this intron is linked to the nucleotide sequence that encodes the S -motif.
Steffen P. Graether - One of the best experts on this subject based on the ideXlab platform.
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in vivo evidence for homo and heterodimeric interactions of arabidopsis thaliana dehydrins atcor47 aterd10 and atrab18
Scientific Reports, 2017Co-Authors: Itzell E Hernandezsanchez, Steffen P. Graether, Israel Marurilopez, Juan Francisco JimenezbremontAbstract:Dehydrins (DHNs) are intrinsically disordered proteins that play central roles in plant abiotic stress responses; however, how they work remains unclear. Herein, we report the in planta subcellular localization of Arabidopsis thaliana DHNs AtCOR47, AtERD10, and AtRAB18 through GFP translational fusions. To explore the dimerization ability of the Arabidopsis acidic DHNs AtCOR47 and AtERD10, we conducted an in planta DHN binding assay using the Bimolecular Fluorescence Complementation (BiFC) technique. Our analyses revealed homodimeric interactions for AtCOR47 and AtERD10; interestingly, heterodimeric associations also occurred with these DHNs, and these interactions were observed in the cytosol of tobacco cells. Furthermore, we evaluated whether Arabidopsis basic DHNs, such as AtRAB18, could also interact with itself and/or with AtCOR47 and AtERD10 in the BiFC system. Our data revealed homodimeric RAB18 complexes in the nucleus and cytosol, while heterodimeric associations between AtRAB18 and acidic DHNs occurred only in the cytosol. Finally, we demonstrated the presence of heterodimeric complexes among Arabidopsis AtCOR47, AtERD10, and AtRAB18 DHNs with their acidic ortholog the OpsDHN1 from Opuntia streptacantha; these heterodimeric interactions showed different subcellular distributions. Our results guide DHN research toward a new scenario where DHN/DHN oligomerization could be explored as a part of their molecular mechanism.
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A dehydrin-dehydrin interaction: the case of SK3 from Opuntia streptacantha
Frontiers in plant science, 2014Co-Authors: Itzell E. Hernández-sánchez, Steffen P. Graether, David M. Martynowicz, Aída Araceli Rodríguez-hernández, Maria B. Pérez-morales, Juan Francisco Jiménez-bremontAbstract:Dehydrins belongs to a large group of highly hydrophilic proteins known as Late Embryogenesis Abundant (LEA) proteins. It is well know that dehydrins are intrinsically disordered plant proteins that accumulate during the late stages of embryogenesis and in response to abiotic stresses; however, the molecular mechanisms by which their functions are carried out are still unclear. We have previously reported that transgenic Arabidopsis plants overexpressing an Opuntia streptacantha SK3 dehydrin (OpsDHN1) show enhanced tolerance to freezing stress. Herein, we show using a split-ubiquitin yeast two-hybrid system that OpsDHN1 dimerizes. We found that the deletion of regions containing K-segments and the histidine-rich region in the OpsDHN1 protein affects dimer formation. Not surprisingly, in silico protein sequence analysis suggests that OpsDHN1 is an intrinsically disordered protein, an observation that was confirmed by circular dichroism and gel filtration of the recombinantly expressed protein. The addition of zinc triggered the association of recombinantly expressed OpsDHN1 protein, likely through its histidine-rich motif. These data brings new insights about the molecular mechanism of the OpsDHN1 SK3-dehydrin
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a dehydrin dehydrin interaction the case of sk3 from Opuntia streptacantha
Frontiers in Plant Science, 2014Co-Authors: Itzell E Hernandezsanchez, Steffen P. Graether, David M. Martynowicz, Aida Araceli Rodriguezhernandez, Maria B Perezmorales, Juan Francisco JimenezbremontAbstract:Dehydrins belongs to a large group of highly hydrophilic proteins known as Late Embryogenesis Abundant (LEA) proteins. It is well know that dehydrins are intrinsically disordered plant proteins that accumulate during the late stages of embryogenesis and in response to abiotic stresses; however, the molecular mechanisms by which their functions are carried out are still unclear. We have previously reported that transgenic Arabidopsis plants overexpressing an Opuntia streptacantha SK3 dehydrin (OpsDHN1) show enhanced tolerance to freezing stress. Herein, we show using a split-ubiquitin yeast two-hybrid system that OpsDHN1 dimerizes. We found that the deletion of regions containing K-segments and the histidine-rich region in the OpsDHN1 protein affects dimer formation. Not surprisingly, in silico protein sequence analysis suggests that OpsDHN1 is an intrinsically disordered protein, an observation that was confirmed by circular dichroism and gel filtration of the recombinantly expressed protein. The addition of zinc triggered the association of recombinantly expressed OpsDHN1 protein, likely through its histidine-rich motif. These data brings new insights about the molecular mechanism of the OpsDHN1 SK3-dehydrin
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División de Biología Molecular,
2014Co-Authors: Opuntia Streptacantha, Juan Francisco Jiménez-bremont, Steffen P. Graether, David M. Martynowicz, Itzell E. Hernández-sánchez, Aída Araceli Rodríguez-hernández, Maria B. Pérez-morales, Yueyun Hong, Huazhong Agricultural, Nabil I. ElsheeryAbstract:Dehydrins belongs to a large group of highly hydrophilic proteins known as Late Embryogenesis Abundant (LEA) proteins. It is well known that dehydrins are intrinsically disordered plant proteins that accumulate during the late stages of embryogenesis and in response to abiotic stresses; however, the molecular mechanisms by which their functions are carried out are still unclear. We have previously reported that transgenic Arabidopsis plants overexpressing an Opuntia streptacantha SK3 dehydrin (OpsDHN1) show enhanced tolerance to freezing stress. Herein, we show using a split-ubiquitin yeast two-hybrid system that OpsDHN1 dimerizes. We found that the deletion of regions containing K-segments and the histidine-rich region in the OpsDHN1 protein affects dimer formation. Not surprisingly, in silico protein sequence analysis suggests that OpsDHN1 is an intrinsically disordered protein, an observation that was confirmed by circular dichroism and gel filtration of the recombinantly expressed protein. The addition of zinc triggered the association of recombinantly expressed OpsDHN1 protein, likely through its histidine-rich motif. These data brings new insights about the molecular mechanism of the OpsDHN
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The Importance of Size and Disorder in the Cryoprotective Effects of Dehydrins
Plant physiology, 2013Co-Authors: Stephanie Hughes, Verena Schart, Janet Malcolmson, Kaley A. Hogarth, David M. Martynowicz, Erik Tralman-baker, Shruti N. Patel, Steffen P. GraetherAbstract:Dehydrins protect plant proteins and membranes from damage during drought and cold. Vitis riparia K2 is a 48-residue protein that can protect lactate dehydrogenase from freeze-thaw damage by preventing the aggregation and denaturation of the enzyme. To further elucidate its mechanism, we used a series of V. riparia K2 concatemers (K4, K6, K8, and K10) and natural dehydrins (V. riparia YSK2, 60 kilodalton peach dehydrin [PCA60], barley dehydrin5 [Dhn5], Thellungiella salsuginea dehydrin2 [TsDHN-2], and Opuntia streptacantha dehydrin1 [OpsDHN-1]) to test the effect of the number of K-segments and dehydrin size on their ability to protect lactate dehydrogenase from freeze-thaw damage. The results show that the larger the hydrodynamic radius of the dehydrin, the more effective the cryoprotection. A similar trend is observed with polyethylene glycol, which would suggest that the protection is simply a nonspecific volume exclusion effect that can be manifested by any protein. However, structured proteins of a similar range of sizes did not show the same pattern and level of cryoprotection. Our results suggest that with respect to enzyme protection, dehydrins function primarily as molecular shields and that their intrinsic disorder is required for them to be an effective cryoprotectant. Lastly, we show that the cryoprotection by a dehydrin is not due to any antifreeze protein-like activity, as has been reported previously.
Alejandro Blanco-labra - One of the best experts on this subject based on the ideXlab platform.
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Characterization of a highly stable trypsin-like proteinase inhibitor from the seeds of Opuntia streptacantha (O. streptacantha Lemaire).
Phytochemistry, 2009Co-Authors: Jorge Ariel Torres-castillo, C Mondragón Jacobo, Alejandro Blanco-labraAbstract:A trypsin inhibitor from Opuntia streptacantha Lemaire (Prickly pear) seeds was purified and characterized. Of several proteases tested, this inhibitor showed specificity to trypsin-like enzymes. The major inhibitor present in these seeds showed distinctive characteristics, most notably a low molecular weight of 4.19 kDa, as determined by MALDI TOF, and an unusually high thermal stability, retaining most of the activity after heating the sample 1h to 120 degrees C with a pressure of 1 kg/cm(2). Its complete amino acid sequence was obtained through mass spectrometry, this establishing presence a blocked N-terminal region. When comparing its sequence in the MEROPS database for peptidases and peptidase inhibitors, it showed 34.48% identity with a serine-proteinase inhibitor from the I15 family.
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Characterization of a highly stable trypsin-like proteinase inhibitor from the seeds of Opuntia streptacantha (O. streptacantha Lemaire).
Phytochemistry, 2009Co-Authors: Jorge Ariel Torres-castillo, C. Mondragón Jacobo, Alejandro Blanco-labraAbstract:Abstract A trypsin inhibitor from Opuntia streptacantha Lemaire (Prickly pear) seeds was purified and characterized. Of several proteases tested, this inhibitor showed specificity to trypsin-like enzymes. The major inhibitor present in these seeds showed distinctive characteristics, most notably a low molecular weight of 4.19 kDa, as determined by MALDI TOF, and an unusually high thermal stability, retaining most of the activity after heating the sample 1 h to 120 °C with a pressure of 1 kg/cm 2 . Its complete amino acid sequence was obtained through mass spectrometry, this establishing presence a blocked N-terminal region. When comparing its sequence in the MEROPS database for peptidases and peptidase inhibitors, it showed 34.48% identity with a serine-proteinase inhibitor from the I15 family.
Anne Negre-salvayre - One of the best experts on this subject based on the ideXlab platform.
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Dietary cladode powder from wild type and domesticated Opuntia species reduces atherogenesis in apoE knock-out mice
Journal of Physiology and Biochemistry, 2016Co-Authors: Sandra Garoby-salom, Caroline Camare, Michel Rossignol, Robert Salvayre, Françoise Guéraud, Ana-paulina Barba De La Rosa, María Del Socorro Santos Díaz, Anne Negre-salvayreAbstract:Dietary intake of Opuntia species may prevent the development of cardiovascular diseases. The present study was designed to characterize the biological antioxidant and anti-inflammatory properties of Opuntia species and to investigate whether Opuntia cladodes prevent the development of atherosclerosis in vivo, in apoE^−KO mice. The effects of the two Opuntia species, the wild Opuntia streptacantha and the domesticated Opuntia ficus-indica , were tested on the generation of intra- and extracellular reactive oxygen species (ROS) production and kinetics of the LDL oxidation by murine CRL2181 endothelial cells and on the subsequent inflammatory signaling leading to the adhesion of monocytes on the activated endothelium and the formation of foam cells. Opuntia species blocked the extracellular ROS (superoxide anion) generation and LDL oxidation by CRL2181, as well as the intracellular ROS rise and signaling evoked by the oxidized LDL, including the nuclear translocation of the transcription factor NFκB, the expression of ICAM-1 and VCAM-1 adhesion molecules, and the adhesion of monocytes to CRL2181. In vivo, Opuntia significantly reduced the formation of atherosclerotic lesions and the accumulation of 4-hydroxynonenal adducts in the vascular wall of apoE-KO mice, indicating that Opuntia cladodes prevent lipid oxidation in the vascular wall. In conclusion, wild and domesticated Opuntia species exhibit antioxidant, anti-inflammatory, and antiatherogenic properties which emphasize their nutritional benefit for preventing cardiovascular diseases.
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Dietary cladode powder from wild type and domesticated Opuntia species reduces atherogenesis in apoE knock-out mice
Journal of Physiology and Biochemistry, 2016Co-Authors: Sandra Garoby-salom, Caroline Camare, Michel Rossignol, Robert Salvayre, Françoise Guéraud, Ana-paulina Barba De La Rosa, María Del Socorro Santos Díaz, Anne Negre-salvayreAbstract:Dietary intake of Opuntia species may prevent the development of cardiovascular diseases. The present study was designed to characterize the biological antioxidant and anti-inflammatory properties of Opuntia species and to investigate whether Opuntia cladodes prevent the development of atherosclerosis in vivo, in apoE(-)KO mice. The effects of the two Opuntia species, the wild Opuntia streptacantha and the domesticated Opuntia ficus-indica, were tested on the generation of intra- and extracellular reactive oxygen species (ROS) production and kinetics of the LDL oxidation by murine CRL2181 endothelial cells and on the subsequent inflammatory signaling leading to the adhesion of monocytes on the activated endothelium and the formation of foam cells. Opuntia species blocked the extracellular ROS (superoxide anion) generation and LDL oxidation by CRL2181, as well as the intracellular ROS rise and signaling evoked by the oxidized LDL, including the nuclear translocation of the transcription factor NF kappa B, the expression of ICAM-1 and VCAM-1 adhesion molecules, and the adhesion of monocytes to CRL2181. In vivo, Opuntia significantly reduced the formation of atherosclerotic lesions and the accumulation of 4-hydroxynonenal adducts in the vascular wall of apoE-KO mice, indicating that Opuntia cladodes prevent lipid oxidation in the vascular wall. In conclusion, wild and domesticated Opuntia species exhibit antioxidant, anti-inflammatory, and antiatherogenic properties which emphasize their nutritional benefit for preventing cardiovascular diseases.
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Establishment of callus and cell suspensions of wild and domesticated Opuntia species: study on their potential as a source of metabolite production
Plant Cell Tissue and Organ Culture, 2016Co-Authors: Marissa Robles-martinez, Michel Rossignol, Françoise Guéraud, Anne Negre-salvayre, Ana Paulina Barba-de La Rosa, Maria Del Socorro Santos-diazAbstract:Opuntia streptacantha and Opuntia megacantha are considered wild species, while Opuntia ficus-indica is deemed a domesticated species. Opuntia species have relevant nutritional and medicinal properties, related to their high levels of phenolics compounds. The objective of this study was to establish callus and cell suspensions of O. streptacantha (cv. Tuna loca), O. megacantha (cv. Rubi reina), and O. ficus-indica (cv. Rojo vigor), and to compare their potential as a source of metabolite production in vitro. Opuntia seeds were physically and chemically scarified obtaining 70-100 % germination. Fast-growing callus and cell suspensions were generated on Murashige and Skoog media supplemented with different concentrations of 2,4-D (0.5, 1.0 mg L-1), and BAP (1.5, 2.0 mg L-1) applied alone or in combination. In general, media containing both, 2,4-D and BAP were more efficient in inducing callus formation than 2,4-D or BAP alone. The content of phenolic acids and flavonoids, and antioxidant activity were all similar in callus and suspensions; however, growth rate was higher in cell suspensions in all species. Production of phenolic acids was correlated to the antioxidant activity, with these parameters being higher in O. streptacantha, the wild species. The concentration of metabolites in callus and cellular suspensions was about three times higher compared with cladodes, indicating that the use of in vitro cultures of Opuntia species is a viable alternative for the production of antioxidants under controlled conditions.