The Experts below are selected from a list of 16329 Experts worldwide ranked by ideXlab platform
Manoj Nath - One of the best experts on this subject based on the ideXlab platform.
-
salt stress triggers augmented levels of na ca2 and ros and alter stress responsive gene expression in roots of cbl9 and cipk23 Knockout Mutants of arabidopsis thaliana
Environmental and Experimental Botany, 2019Co-Authors: Manoj Nath, Deepesh Bhatt, Ajay N. Jain, Saurabh C. Saxena, Shabnam K. Saifi, Sandep Yadav, Manisha Negi, Ram Prasad, Narendra TutejaAbstract:Abstract Salinity stress adversely affects plant growth and development. Calcineurin B-like proteins (CBLs) are a family of plant calcium sensors that relay signals by interacting with a family of CBL-interacting protein kinases (CIPKs). The current work entails a reverse genetic approach for deciphering the roles of CBL9 and CIPK23 in the roots of Arabidopsis thaliana (Arabidopsis) during salinity stress. The Knockout Mutants cbl9 and cipk23 showed salt sensitivity and significantly higher levels of sodium ion (Na+), cytosolic calcium [Ca2+]cyt, and reactive oxygen species (ROS) and an increased death of the cells in root tips compared to the wild-type (WT) under salinity stress (100 mM NaCl). Further, we intended to decipher the effect of salinity stress on the relative expression levels of few key genes that play pivotal role in salinity stress responses and ROS-scavenging activities in the roots of the WT and the mutant seedlings. Compared with the WT, the relative expression levels of the genes in cbl9 and/or cipk23 were significantly augmented (CAX1, 2 and 4, ACA1, CNGC14 and 19, 5PTase2 and DHAR2), attenuated (SOS1, APX1, ALX8, CAT2 and 3, DHAR1, SQD1 and 2) and remained unaffected (NHX1–6 and 8 and GST11). These findings were further corroborated with the increased level of stress markers namely Hydrogen peroxide (H2O2) and Malondialdehyde (MDA) and reduction in chlorophyll, biomass and ascorbate peroxidase (APX) activity in Mutants under salt stress condition. The results thus highlighted pivotal roles of CBL9 and CIPK23 on salinity stress responses and ROS-scavenging activities in the roots during salinity stress and a schematic model is presented.
-
Salt stress triggers augmented levels of Na+, Ca2+ and ROS and alter stress-responsive gene expression in roots of CBL9 and CIPK23 Knockout Mutants of Arabidopsis thaliana
Environmental and Experimental Botany, 2019Co-Authors: Manoj Nath, Deepesh Bhatt, Ajay N. Jain, Saurabh C. Saxena, Shabnam K. Saifi, Sandep Yadav, Manisha Negi, Ram Prasad, Narendra TutejaAbstract:Abstract Salinity stress adversely affects plant growth and development. Calcineurin B-like proteins (CBLs) are a family of plant calcium sensors that relay signals by interacting with a family of CBL-interacting protein kinases (CIPKs). The current work entails a reverse genetic approach for deciphering the roles of CBL9 and CIPK23 in the roots of Arabidopsis thaliana (Arabidopsis) during salinity stress. The Knockout Mutants cbl9 and cipk23 showed salt sensitivity and significantly higher levels of sodium ion (Na+), cytosolic calcium [Ca2+]cyt, and reactive oxygen species (ROS) and an increased death of the cells in root tips compared to the wild-type (WT) under salinity stress (100 mM NaCl). Further, we intended to decipher the effect of salinity stress on the relative expression levels of few key genes that play pivotal role in salinity stress responses and ROS-scavenging activities in the roots of the WT and the mutant seedlings. Compared with the WT, the relative expression levels of the genes in cbl9 and/or cipk23 were significantly augmented (CAX1, 2 and 4, ACA1, CNGC14 and 19, 5PTase2 and DHAR2), attenuated (SOS1, APX1, ALX8, CAT2 and 3, DHAR1, SQD1 and 2) and remained unaffected (NHX1–6 and 8 and GST11). These findings were further corroborated with the increased level of stress markers namely Hydrogen peroxide (H2O2) and Malondialdehyde (MDA) and reduction in chlorophyll, biomass and ascorbate peroxidase (APX) activity in Mutants under salt stress condition. The results thus highlighted pivotal roles of CBL9 and CIPK23 on salinity stress responses and ROS-scavenging activities in the roots during salinity stress and a schematic model is presented.
Narendra Tuteja - One of the best experts on this subject based on the ideXlab platform.
-
salt stress triggers augmented levels of na ca2 and ros and alter stress responsive gene expression in roots of cbl9 and cipk23 Knockout Mutants of arabidopsis thaliana
Environmental and Experimental Botany, 2019Co-Authors: Manoj Nath, Deepesh Bhatt, Ajay N. Jain, Saurabh C. Saxena, Shabnam K. Saifi, Sandep Yadav, Manisha Negi, Ram Prasad, Narendra TutejaAbstract:Abstract Salinity stress adversely affects plant growth and development. Calcineurin B-like proteins (CBLs) are a family of plant calcium sensors that relay signals by interacting with a family of CBL-interacting protein kinases (CIPKs). The current work entails a reverse genetic approach for deciphering the roles of CBL9 and CIPK23 in the roots of Arabidopsis thaliana (Arabidopsis) during salinity stress. The Knockout Mutants cbl9 and cipk23 showed salt sensitivity and significantly higher levels of sodium ion (Na+), cytosolic calcium [Ca2+]cyt, and reactive oxygen species (ROS) and an increased death of the cells in root tips compared to the wild-type (WT) under salinity stress (100 mM NaCl). Further, we intended to decipher the effect of salinity stress on the relative expression levels of few key genes that play pivotal role in salinity stress responses and ROS-scavenging activities in the roots of the WT and the mutant seedlings. Compared with the WT, the relative expression levels of the genes in cbl9 and/or cipk23 were significantly augmented (CAX1, 2 and 4, ACA1, CNGC14 and 19, 5PTase2 and DHAR2), attenuated (SOS1, APX1, ALX8, CAT2 and 3, DHAR1, SQD1 and 2) and remained unaffected (NHX1–6 and 8 and GST11). These findings were further corroborated with the increased level of stress markers namely Hydrogen peroxide (H2O2) and Malondialdehyde (MDA) and reduction in chlorophyll, biomass and ascorbate peroxidase (APX) activity in Mutants under salt stress condition. The results thus highlighted pivotal roles of CBL9 and CIPK23 on salinity stress responses and ROS-scavenging activities in the roots during salinity stress and a schematic model is presented.
-
Salt stress triggers augmented levels of Na+, Ca2+ and ROS and alter stress-responsive gene expression in roots of CBL9 and CIPK23 Knockout Mutants of Arabidopsis thaliana
Environmental and Experimental Botany, 2019Co-Authors: Manoj Nath, Deepesh Bhatt, Ajay N. Jain, Saurabh C. Saxena, Shabnam K. Saifi, Sandep Yadav, Manisha Negi, Ram Prasad, Narendra TutejaAbstract:Abstract Salinity stress adversely affects plant growth and development. Calcineurin B-like proteins (CBLs) are a family of plant calcium sensors that relay signals by interacting with a family of CBL-interacting protein kinases (CIPKs). The current work entails a reverse genetic approach for deciphering the roles of CBL9 and CIPK23 in the roots of Arabidopsis thaliana (Arabidopsis) during salinity stress. The Knockout Mutants cbl9 and cipk23 showed salt sensitivity and significantly higher levels of sodium ion (Na+), cytosolic calcium [Ca2+]cyt, and reactive oxygen species (ROS) and an increased death of the cells in root tips compared to the wild-type (WT) under salinity stress (100 mM NaCl). Further, we intended to decipher the effect of salinity stress on the relative expression levels of few key genes that play pivotal role in salinity stress responses and ROS-scavenging activities in the roots of the WT and the mutant seedlings. Compared with the WT, the relative expression levels of the genes in cbl9 and/or cipk23 were significantly augmented (CAX1, 2 and 4, ACA1, CNGC14 and 19, 5PTase2 and DHAR2), attenuated (SOS1, APX1, ALX8, CAT2 and 3, DHAR1, SQD1 and 2) and remained unaffected (NHX1–6 and 8 and GST11). These findings were further corroborated with the increased level of stress markers namely Hydrogen peroxide (H2O2) and Malondialdehyde (MDA) and reduction in chlorophyll, biomass and ascorbate peroxidase (APX) activity in Mutants under salt stress condition. The results thus highlighted pivotal roles of CBL9 and CIPK23 on salinity stress responses and ROS-scavenging activities in the roots during salinity stress and a schematic model is presented.
Thomas Efferth - One of the best experts on this subject based on the ideXlab platform.
-
ursolic acid ameliorates stress and reactive oxygen species in c elegans Knockout Mutants by the dopamine dop1 and dop3 receptors
Phytomedicine, 2021Co-Authors: Janine Nas, Thomas EfferthAbstract:Abstract Background: Depression and stress-related disorders are leading causes of death worldwide. Standard treatments elevating serotonin or noradrenaline levels are not sufficiently effective and cause adverse side effects. A connection between dopamine pathways and stress-related disorders has been suggested. Compounds derived from herbal medicine could be a promising alternative. We examined the neuroprotective effects of ursolic acid (UA) by focusing on dopamine signalling. Methods: Trolox equivalent capacity assay was used to determine the antioxidant activities of UA in vitro. C. elegans N2 wildtype and dopamine receptor-Knockout Mutants (dop1-deficient RB665 and dop3-deficient LX703 strains) were used as in vivo models. H2DCFDA and acute juglone assays were applied to determine the antioxidant activity in dependency of dopamine pathways in vivo. Stress was assessed by heat and acute osmotic stress assays. The influence of UA on overall survival was analyzed by a life span assay. The dop1 and dop3 mRNA expression was determined by real time RT-PCR. We also examined the binding affinity of UA towards C. elegans Dop1 and Dop3 receptors as well as human dopamine receptors D1 and D3 by molecular docking. Results: Antioxidant activity assays showed that UA exerts strong antioxidant activity. UA increased resistance towards oxidative, osmotic and heat stress. Additionally, UA increased life span of nematodes. Moreover, dop1 and dop3 gene expression was significantly enhanced upon UA treatment. Docking analysis revealed stronger binding affinity of UA to C. elegans and human dopamine receptors than the natural ligand, dopamine. Binding to Dop1 was stronger than to Dop3. Conclusion: UA reduced stress-dependent ROS generation and acted through Dop1 and to a lesser extent through Dop3 to reduce stress and prolong life span in C. elegans. These results indicate that UA could be a promising lead compound for the development of new antidepressant medications.
-
Ursolic acid ameliorates stress and reactive oxygen species in C. elegans Knockout Mutants by the dopamine Dop1 and Dop3 receptors.
Phytomedicine : international journal of phytotherapy and phytopharmacology, 2020Co-Authors: Janine Naß, Thomas EfferthAbstract:Depression and stress-related disorders are leading causes of death worldwide. Standard treatments elevating serotonin or noradrenaline levels are not sufficiently effective and cause adverse side effects. A connection between dopamine pathways and stress-related disorders has been suggested. Compounds derived from herbal medicine could be a promising alternative. We examined the neuroprotective effects of ursolic acid (UA) by focusing on dopamine signalling. Trolox equivalent capacity assay was used to determine the antioxidant activities of UA in vitro. C. elegans N2 wildtype and dopamine receptor-Knockout Mutants (dop1-deficient RB665 and dop3-deficient LX703 strains) were used as in vivo models. H2DCFDA and acute juglone assays were applied to determine the antioxidant activity in dependency of dopamine pathways in vivo. Stress was assessed by heat and acute osmotic stress assays. The influence of UA on overall survival was analyzed by a life span assay. The dop1 and dop3 mRNA expression was determined by real time RT-PCR. We also examined the binding affinity of UA towards C. elegans Dop1 and Dop3 receptors as well as human dopamine receptors D1 and D3 by molecular docking. Antioxidant activity assays showed that UA exerts strong antioxidant activity. UA increased resistance towards oxidative, osmotic and heat stress. Additionally, UA increased life span of nematodes. Moreover, dop1 and dop3 gene expression was significantly enhanced upon UA treatment. Docking analysis revealed stronger binding affinity of UA to C. elegans and human dopamine receptors than the natural ligand, dopamine. Binding to Dop1 was stronger than to Dop3. UA reduced stress-dependent ROS generation and acted through Dop1 and to a lesser extent through Dop3 to reduce stress and prolong life span in C. elegans. These results indicate that UA could be a promising lead compound for the development of new antidepressant medications. Copyright © 2020. Published by Elsevier GmbH.
John B Dame - One of the best experts on this subject based on the ideXlab platform.
-
Effects on growth, hemoglobin metabolism and paralogous gene expression resulting from disruption of genes encoding the digestive vacuole plasmepsins of Plasmodium falciparum.
International journal for parasitology, 2006Co-Authors: J Alfredo Bonilla, Pedro A Moura, Tonya D Bonilla, Charles A Yowell, David A Fidock, John B DameAbstract:Four of the plasmepsins of Plasmodium falciparum are localised in the digestive vacuole (DV) of the asexual blood stage parasite (PfPM1, PfPM2, PfPM4 and PfHAP), and each of these aspartic proteinases has been successfully targeted by gene disruption. This study describes further characterisation of the single-plasmepsin Knockout Mutants, and the creation and characterisation of double-plasmepsin Knockout Mutants lacking complete copies of pfpm2 and pfpm1 or pfhap and pfpm2. Double-plasmepsin Knockout Mutants were created by transfecting pre-existing Knockout Mutants with a second plasmid Knockout construct. PCR and Southern blot analysis demonstrate the integration of a large concatamer of each plasmid construct into the targeted gene. All Mutants have been characterised to assess the involvement of the DV plasmepsins in sustaining growth during the asexual blood stage. Analyses reaffirmed that Knockout Mutants Deltapfpm1 and Deltapfpm4 had lower replication rates in the asexual erythrocytic stage than the parental line (Dd2), but double-plasmepsin Knockout Mutants lacking intact copies of either pfpm2 and pfpm1, or pfpm2 and pfhap, had normal growth rates compared with Dd2. The amount of crystalline hemozoin produced per parasite during the asexual cycle was measured in each single-plasmepsin Knockout to estimate the effect of each DV plasmepsin on hemoglobin digestion. Only Deltapfpm4 had a statistically significant reduction in hemozoin accumulation, indicating that hemoglobin digestion was impaired in this mutant. In the single-plasmepsin Knockouts, no statistically significant differences were found in the steady state levels of mRNA from the remaining intact DV plasmepsin genes. Disruption of a DV plasmepsin gene does not affect the accumulation of mRNA encoding the remaining paralogous plasmepsins, and Western blot analysis confirmed that the accumulation of the paralogous plasmepsins in each Knockout mutant was similar among all clones examined.
-
Effects on growth, hemoglobin metabolism and paralogous gene expression resulting from disruption of genes encoding the digestive vacuole plasmepsins of Plasmodium falciparum
International Journal for Parasitology, 2006Co-Authors: J Alfredo Bonilla, Pedro A Moura, Tonya D Bonilla, Charles A Yowell, David A Fidock, John B DameAbstract:Abstract Four of the plasmepsins of Plasmodium falciparum are localised in the digestive vacuole (DV) of the asexual blood stage parasite (PfPM1, PfPM2, PfPM4 and PfHAP), and each of these aspartic proteinases has been successfully targeted by gene disruption. This study describes further characterisation of the single-plasmepsin Knockout Mutants, and the creation and characterisation of double-plasmepsin Knockout Mutants lacking complete copies of pfpm 2 and pfpm 1 or pfhap and pfpm 2. Double-plasmepsin Knockout Mutants were created by transfecting pre-existing Knockout Mutants with a second plasmid Knockout construct. PCR and Southern blot analysis demonstrate the integration of a large concatamer of each plasmid construct into the targeted gene. All Mutants have been characterised to assess the involvement of the DV plasmepsins in sustaining growth during the asexual blood stage. Analyses reaffirmed that Knockout Mutants Δ pfpm 1 and Δ pfpm 4 had lower replication rates in the asexual erythrocytic stage than the parental line (Dd2), but double-plasmepsin Knockout Mutants lacking intact copies of either pfpm 2 and pfpm 1, or pfpm2 and pfhap , had normal growth rates compared with Dd2. The amount of crystalline hemozoin produced per parasite during the asexual cycle was measured in each single-plasmepsin Knockout to estimate the effect of each DV plasmepsin on hemoglobin digestion. Only Δ pfpm 4 had a statistically significant reduction in hemozoin accumulation, indicating that hemoglobin digestion was impaired in this mutant. In the single-plasmepsin Knockouts, no statistically significant differences were found in the steady state levels of mRNA from the remaining intact DV plasmepsin genes. Disruption of a DV plasmepsin gene does not affect the accumulation of mRNA encoding the remaining paralogous plasmepsins, and Western blot analysis confirmed that the accumulation of the paralogous plasmepsins in each Knockout mutant was similar among all clones examined.
Hirotada Mori - One of the best experts on this subject based on the ideXlab platform.
-
Systematic phenome analysis of Escherichia coli multiple‐Knockout Mutants reveals hidden reactions in central carbon metabolism
Molecular systems biology, 2009Co-Authors: Kenji Nakahigashi, Yoshihiro Toya, Nobuyoshi Ishii, Tomoyoshi Soga, Miki Hasegawa, Hisami Watanabe, Yuki Takai, Masayuki Honma, Hirotada Mori, Masaru TomitaAbstract:Central carbon metabolism is a basic and exhaustively analyzed pathway. However, the intrinsic robustness of the pathway might still conceal uncharacterized reactions. To test this hypothesis, we constructed systematic multiple-Knockout Mutants involved in central carbon catabolism in Escherichia coli and tested their growth under 12 different nutrient conditions. Differences between in silico predictions and experimental growth indicated that unreported reactions existed within this extensively analyzed metabolic network. These putative reactions were then confirmed by metabolome analysis and in vitro enzymatic assays. Novel reactions regarding the breakdown of sedoheptulose-7-phosphate to erythrose-4-phosphate and dihydroxyacetone phosphate were observed in transaldolase-deficient Mutants, without any noticeable changes in gene expression. These reactions, triggered by an accumulation of sedoheptulose-7-phosphate, were catalyzed by the universally conserved glycolytic enzymes ATP-dependent phosphofructokinase and aldolase. The emergence of an alternative pathway not requiring any changes in gene expression, but rather relying on the accumulation of an intermediate metabolite may be a novel mechanism mediating the robustness of these metabolic networks.
-
systematic phenome analysis of escherichia coli multiple Knockout Mutants reveals hidden reactions in central carbon metabolism
Molecular Systems Biology, 2009Co-Authors: Kenji Nakahigashi, Yoshihiro Toya, Nobuyoshi Ishii, Tomoyoshi Soga, Miki Hasegawa, Hisami Watanabe, Yuki Takai, Masayuki Honma, Hirotada MoriAbstract:Central carbon metabolism is a basic and exhaustively analyzed pathway. However, the intrinsic robustness of the pathway might still conceal uncharacterized reactions. To test this hypothesis, we constructed systematic multiple-Knockout Mutants involved in central carbon catabolism in Escherichia coli and tested their growth under 12 different nutrient conditions. Differences between in silico predictions and experimental growth indicated that unreported reactions existed within this extensively analyzed metabolic network. These putative reactions were then confirmed by metabolome analysis and in vitro enzymatic assays. Novel reactions regarding the breakdown of sedoheptulose-7-phosphate to erythrose-4-phosphate and dihydroxyacetone phosphate were observed in transaldolase-deficient Mutants, without any noticeable changes in gene expression. These reactions, triggered by an accumulation of sedoheptulose-7-phosphate, were catalyzed by the universally conserved glycolytic enzymes ATP-dependent phosphofructokinase and aldolase. The emergence of an alternative pathway not requiring any changes in gene expression, but rather relying on the accumulation of an intermediate metabolite may be a novel mechanism mediating the robustness of these metabolic networks.
-
construction of escherichia coli k 12 in frame single gene Knockout Mutants the keio collection
Molecular Systems Biology, 2006Co-Authors: Miki Hasegawa, Yuki Takai, Masaru Tomita, Tomoya Baba, Yoshiko Okumura, Miki Baba, Kirill A Datsenko, Barry L Wanner, Hirotada MoriAbstract:We have systematically made a set of precisely defined, single‐gene deletions of all nonessential genes in Escherichia coli K‐12. Open‐reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one‐step method for inactivation of chromosomal genes and primers designed to create in‐frame deletions upon excision of the resistance cassette. Of 4288 genes targeted, Mutants were obtained for 3985. To alleviate problems encountered in high‐throughput studies, two independent Mutants were saved for every deleted gene. These Mutants—the ‘Keio collection’—provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome‐wide testing of mutational effects in a common strain background, E . coli K‐12 BW25113. We were unable to disrupt 303 genes, including 37 of unknown function, which are candidates for essential genes. Distribution is being handled via GenoBase (http://ecoli.aist‐nara.ac.jp/).
