This study was designed to analyze the effect of the mitochondrial respiratory pathways of Candida albicans （C. albicans） on the biofilm formation. The 2, 3-bis （2-methoxy- 4-nitro-5-sulfophenyl）-2H-tetrazolium-5-...This study was designed to analyze the effect of the mitochondrial respiratory pathways of Candida albicans （C. albicans） on the biofilm formation. The 2, 3-bis （2-methoxy- 4-nitro-5-sulfophenyl）-2H-tetrazolium-5-carboxanilide （XTT） reduction assay was used to measure the metabolic activities of biofilms formed by the C. albicans which were cultured in the presence of respiratory pathways inhibitors. The biofilms formed by the wide type （WT）, GOA1-deleted （GOA31）, GOAl-reconstituted （GOA32）, AOXla-deleted （AOX1） and AOX1b- deleted （AOX2） C. albicans strains were examined by the XTT reduction assay and fluorescence microscopy. The expression of adhesion-related genes BCR1, ALS1, ALS3, ECEI and HWP1 in the biofilms formed by the above five C. albicans strains was detected by real time polymerase chain reaction. It was found that the metabolic activity of biofilms formed by C. albicans was decreased in the presence of alternative oxidase inhibitor whereas it was increased in the presence of classical mitochondrial respiratory pathway complex Ⅲ or complex IV inhibitor. AOX1 strain produced scarce biofilms interspersed with few hyphal filaments. Moreover, no significant changes in the expression of BCR1 and ALS3 were observed in the AOX 1 strain, but the expression of ALS1 and ECE1 was down-regulated, and that of HWP1 was up-regulated. These results indicate that both AOX1 and AOX2 can promote the biofilm formation. However, AOX1a primarily plays a regulatory role in biofilm formation in the absence of inducers where the promoting effect is mainly achieved by promoting mycelial formation.展开更多
Previous work illustrated that glucose oxidase (GOx) could be deposited on conducting substrates using asymmetrical alternating current electrophoretic deposition (AC-EPD) to form thick enzyme layers suitable for the ...Previous work illustrated that glucose oxidase (GOx) could be deposited on conducting substrates using asymmetrical alternating current electrophoretic deposition (AC-EPD) to form thick enzyme layers suitable for the manufacturing of highly active biosensors. Here, we modeled the amperometric response of GOx layers to glucose as a function of the thickness of the enzyme layer. The model is based on reaction-diffusion equations with irreversible first-order catalytic reactions. The numerical results displayed qualitative and reasonable quantitative agreement with the experimental data obtained for oxidation currents due to glucose, which increase with the enzyme thickness.展开更多
With the exception of rice, little is known about the existence of respiratory burst oxidase homolog （rboh） gene in cereals. The present study reports the cloning and analysis of a novel rboh gene, termed ZmrbohB, f...With the exception of rice, little is known about the existence of respiratory burst oxidase homolog （rboh） gene in cereals. The present study reports the cloning and analysis of a novel rboh gene, termed ZmrbohB, from maize （Zea mays L.）. The full-length cDNA of ZmrbohB encodes a 942 amino acid protein containing all of the respiratory burst oxidase homolog catalytically critical motifs. Alternative splicing of ZmrbohB has generated two transcript isoforms, ZmrbohB-α and -β. Spliced transcript ZmrbohB-β retains an unspliced intron 11 that carries a premature termination codon and probably leads to nonsense-mediated mRNA decay. Expression analysis showed that two splice isoforms were differentially expressed in various tissues and at different developmental stages, and the major product was ZmrbohB-e. The transcripts of ZmrbohB-α accumulated markedly when the maize seedlings were subjected to various abiotic stimuli, such as wounding, cold （4℃）, heat （40℃）, UV and salinity stress. In addition, several abiotic stimuli also affected the alternative splicing pattern of ZmrbohB except wounding. These results provide new insight into roles in the expression regulation of plant rboh genes and suggest that ZrnrbohB gene may play a role in response to environmental stresses.展开更多
An increase in ultraviolet （UV） B radiation on the earth＇s surface is a feature of current global climate changes. It has been reported that alternative oxidase （AOX） may have a protective role against oxidative ...An increase in ultraviolet （UV） B radiation on the earth＇s surface is a feature of current global climate changes. It has been reported that alternative oxidase （AOX） may have a protective role against oxidative stress induced by environmental stresses, such as UV-B. To better understand the characteristic tolerance of plants to UV-B radiation, the effects of enhanced UV-B radiation on the activity and expression of AOX in red kidney bean （Phaseolus vulgaris） leaves were investigated in the present study. The results show that the total respiration rate and AOX activity in red kidney bean leaves increased significantly during treatment with enhanced UV-B. However, cytochrome oxidase （COX） activity did not change significantly. The H2O2 content was also markedly increased and reached a maximum of 4.45 mmol·L^-1·g^-1 DW （dry weight） at 24 h of UV-B treatment, before dropping rapidly. Both alternative pathway content and alternative pathway activity were increased in the presence of exogenous H2O2. Immunoblotting analysis with anti-AOX monoclonal antibody revealed that expression of the AOX protein increased in red kidney bean leaves under enhanced UV-B radiation, reaching a peak at 72 h. In addition, AOX expression in red kidney bean leaves was induced by exogenous H2O2. These data indicate that the increase in AOX activity in red kidney bean leaves under enhanced UV-B radiation was mainly due to H2O2-induced AOX expression.展开更多
Although mitochondrial alternative oxidase(AOX)has been proposed to play essential roles in high light stress tolerance,the effects of AOX on chlorophyll synthesis are unclear.Previous studies indicated that during gr...Although mitochondrial alternative oxidase(AOX)has been proposed to play essential roles in high light stress tolerance,the effects of AOX on chlorophyll synthesis are unclear.Previous studies indicated that during greening,chlorophyll accumulation was largely delayed in plants whose mitochondrial cyanide-resistant respiration was inhibited by knocking out nuclear encoded AOX gene.Here we show that this delay of chlorophyll accumulation was more significant under high light condition.Inhibition of cyanide-resistant respiration was also accompanied by the increase of plastid NADPH/NADP^+ratio,especially under high light treatment which subsequently blocked the import of multiple plastidial proteins,such as some components of the photosynthetic electron transport chain,the Calvin-Benson cycle enzymes and malate/oxaloacetate shuttle components.Over expression of AOXla rescued the aoxla mutant phenotype,including the chlorophyll accumulation during greening and plastidial protein import.It thus suggests that light intensity affects chlorophyll synthesis during greening process by a metabolic signal,the AOX-derived plastidial NADPH/NADP^+ratio change.And our results thus revealed a molecular mechanism of chloroplast-mitochondria interactions.展开更多
The vital function of mitochondrial alternative oxidase(AOX) pathway in optimizing photosynthesis during plant de-etiolation has been well recognized. However, whether and how AOX impacts the chloroplast biogenesis in...The vital function of mitochondrial alternative oxidase(AOX) pathway in optimizing photosynthesis during plant de-etiolation has been well recognized. However, whether and how AOX impacts the chloroplast biogenesis in algal cells remains unclear. In the present study, the role of AOX in regulating the reassembly of chloroplast in algal cells was investigated by treating Auxenochlorella protothecoides with salicylhydroxamic acid(SHAM), the specific inhibitor to AOX, in the heterotrophy to autotrophy transition process. Several lines of evidences including delayed chlorophyll accumulation, lagged reorganization of chloroplast structure, altered PSI/PSII stoichiometry, and declined photosynthetic activities in SHAM treated cells indicated that the impairment in AOX activity dramatically hindered the development of functioning chloroplast in algal cells. Besides, the cellular ROS levels and activities of antioxidant enzymes were increased by SHAM treatment, and the perturbation on the balance of NAD+/NADH and NADP+/NADPH ratios was also observed in A. protothecoides lacking AOX activity, indicating that AOX was essential in promoting ROS scavenging and keeping the redox homeostasis for algal chloroplast development during greening. Overall, our study revealed the essentiality of mitochondrial AOX pathway in sustaining algal photosynthetic performance and provided novel insights into the physiological roles of AOX on the biogenesis of photosynthetic organelle in algae.展开更多
基金This study was supported by the National Natural Science Foundation of China （No. 81371785）.
文摘This study was designed to analyze the effect of the mitochondrial respiratory pathways of Candida albicans （C. albicans） on the biofilm formation. The 2, 3-bis （2-methoxy- 4-nitro-5-sulfophenyl）-2H-tetrazolium-5-carboxanilide （XTT） reduction assay was used to measure the metabolic activities of biofilms formed by the C. albicans which were cultured in the presence of respiratory pathways inhibitors. The biofilms formed by the wide type （WT）, GOA1-deleted （GOA31）, GOAl-reconstituted （GOA32）, AOXla-deleted （AOX1） and AOX1b- deleted （AOX2） C. albicans strains were examined by the XTT reduction assay and fluorescence microscopy. The expression of adhesion-related genes BCR1, ALS1, ALS3, ECEI and HWP1 in the biofilms formed by the above five C. albicans strains was detected by real time polymerase chain reaction. It was found that the metabolic activity of biofilms formed by C. albicans was decreased in the presence of alternative oxidase inhibitor whereas it was increased in the presence of classical mitochondrial respiratory pathway complex Ⅲ or complex IV inhibitor. AOX1 strain produced scarce biofilms interspersed with few hyphal filaments. Moreover, no significant changes in the expression of BCR1 and ALS3 were observed in the AOX 1 strain, but the expression of ALS1 and ECE1 was down-regulated, and that of HWP1 was up-regulated. These results indicate that both AOX1 and AOX2 can promote the biofilm formation. However, AOX1a primarily plays a regulatory role in biofilm formation in the absence of inducers where the promoting effect is mainly achieved by promoting mycelial formation.
文摘Previous work illustrated that glucose oxidase (GOx) could be deposited on conducting substrates using asymmetrical alternating current electrophoretic deposition (AC-EPD) to form thick enzyme layers suitable for the manufacturing of highly active biosensors. Here, we modeled the amperometric response of GOx layers to glucose as a function of the thickness of the enzyme layer. The model is based on reaction-diffusion equations with irreversible first-order catalytic reactions. The numerical results displayed qualitative and reasonable quantitative agreement with the experimental data obtained for oxidation currents due to glucose, which increase with the enzyme thickness.
基金Supported by the State Key Basic Research and Development Plan of China (2003CB114302)the National Natural Science Foundation of China (30571122).
文摘With the exception of rice, little is known about the existence of respiratory burst oxidase homolog （rboh） gene in cereals. The present study reports the cloning and analysis of a novel rboh gene, termed ZmrbohB, from maize （Zea mays L.）. The full-length cDNA of ZmrbohB encodes a 942 amino acid protein containing all of the respiratory burst oxidase homolog catalytically critical motifs. Alternative splicing of ZmrbohB has generated two transcript isoforms, ZmrbohB-α and -β. Spliced transcript ZmrbohB-β retains an unspliced intron 11 that carries a premature termination codon and probably leads to nonsense-mediated mRNA decay. Expression analysis showed that two splice isoforms were differentially expressed in various tissues and at different developmental stages, and the major product was ZmrbohB-e. The transcripts of ZmrbohB-α accumulated markedly when the maize seedlings were subjected to various abiotic stimuli, such as wounding, cold （4℃）, heat （40℃）, UV and salinity stress. In addition, several abiotic stimuli also affected the alternative splicing pattern of ZmrbohB except wounding. These results provide new insight into roles in the expression regulation of plant rboh genes and suggest that ZrnrbohB gene may play a role in response to environmental stresses.
基金the Doctoral Program of Higher Education of China(20050730017)Foundation of Science and Technology of Gansu Provlnce(3ZS051-A25-018).
文摘An increase in ultraviolet （UV） B radiation on the earth＇s surface is a feature of current global climate changes. It has been reported that alternative oxidase （AOX） may have a protective role against oxidative stress induced by environmental stresses, such as UV-B. To better understand the characteristic tolerance of plants to UV-B radiation, the effects of enhanced UV-B radiation on the activity and expression of AOX in red kidney bean （Phaseolus vulgaris） leaves were investigated in the present study. The results show that the total respiration rate and AOX activity in red kidney bean leaves increased significantly during treatment with enhanced UV-B. However, cytochrome oxidase （COX） activity did not change significantly. The H2O2 content was also markedly increased and reached a maximum of 4.45 mmol·L^-1·g^-1 DW （dry weight） at 24 h of UV-B treatment, before dropping rapidly. Both alternative pathway content and alternative pathway activity were increased in the presence of exogenous H2O2. Immunoblotting analysis with anti-AOX monoclonal antibody revealed that expression of the AOX protein increased in red kidney bean leaves under enhanced UV-B radiation, reaching a peak at 72 h. In addition, AOX expression in red kidney bean leaves was induced by exogenous H2O2. These data indicate that the increase in AOX activity in red kidney bean leaves under enhanced UV-B radiation was mainly due to H2O2-induced AOX expression.
基金supported by the National Nature Science Foundation of China(31070210,91017004)the Doctoral Foundation of the Ministry of Education(20110181110059 and 20120181130008)Sichuan and Chengdu Nature Science Foundation(2010JQ0080,11DXYB097JH-027 and2012JY0078)
文摘Although mitochondrial alternative oxidase(AOX)has been proposed to play essential roles in high light stress tolerance,the effects of AOX on chlorophyll synthesis are unclear.Previous studies indicated that during greening,chlorophyll accumulation was largely delayed in plants whose mitochondrial cyanide-resistant respiration was inhibited by knocking out nuclear encoded AOX gene.Here we show that this delay of chlorophyll accumulation was more significant under high light condition.Inhibition of cyanide-resistant respiration was also accompanied by the increase of plastid NADPH/NADP^+ratio,especially under high light treatment which subsequently blocked the import of multiple plastidial proteins,such as some components of the photosynthetic electron transport chain,the Calvin-Benson cycle enzymes and malate/oxaloacetate shuttle components.Over expression of AOXla rescued the aoxla mutant phenotype,including the chlorophyll accumulation during greening and plastidial protein import.It thus suggests that light intensity affects chlorophyll synthesis during greening process by a metabolic signal,the AOX-derived plastidial NADPH/NADP^+ratio change.And our results thus revealed a molecular mechanism of chloroplast-mitochondria interactions.
基金the National Key Research and Development Project for Synthetic Biology (2018YFA0902500)the National Natural Science Foundation of China (41876188)+1 种基金Shenzhen Grant Plan for Science and Technology (Shenzhen Basic Research Projects: JCYJ20180507182405562)Grant Plan for Demonstration Project for Marine Economic Development in Shenzhen to Dr. Zhangli HU。
文摘The vital function of mitochondrial alternative oxidase(AOX) pathway in optimizing photosynthesis during plant de-etiolation has been well recognized. However, whether and how AOX impacts the chloroplast biogenesis in algal cells remains unclear. In the present study, the role of AOX in regulating the reassembly of chloroplast in algal cells was investigated by treating Auxenochlorella protothecoides with salicylhydroxamic acid(SHAM), the specific inhibitor to AOX, in the heterotrophy to autotrophy transition process. Several lines of evidences including delayed chlorophyll accumulation, lagged reorganization of chloroplast structure, altered PSI/PSII stoichiometry, and declined photosynthetic activities in SHAM treated cells indicated that the impairment in AOX activity dramatically hindered the development of functioning chloroplast in algal cells. Besides, the cellular ROS levels and activities of antioxidant enzymes were increased by SHAM treatment, and the perturbation on the balance of NAD+/NADH and NADP+/NADPH ratios was also observed in A. protothecoides lacking AOX activity, indicating that AOX was essential in promoting ROS scavenging and keeping the redox homeostasis for algal chloroplast development during greening. Overall, our study revealed the essentiality of mitochondrial AOX pathway in sustaining algal photosynthetic performance and provided novel insights into the physiological roles of AOX on the biogenesis of photosynthetic organelle in algae.
