Project description:We were awarded a BBSRC grant about a year ago to undertake some affymetrix gene chip profiling of light and CO2 systemic signalling in Arabidopsis. The design of the proposed experiment is given below and the appropriate funding has been provided by the BBSRC. The aim of the project is to identify the temporal profile of those genes that respond to light and CO2 systemic signals in developing leaves. Moreover, as thes two signals have opposing effects on leaf development to ascertain whether they involve similar or parallel signalling pathways. The experiment is to examine the effect of exposing mature leaves to high CO2 or low light or both on the gene expression profile of developing leaves. We already have data for maize that changes in gene expression profile occur within 4h and that there are a variety of temporal responses that differ between individual gene transcripts. We have also demonstrated that Arabidopsis leaf development is altered by these systemmic signals and that lesions in the jasmonate and ethylene signalling pathways block these responses. Our experimental design is shown below: We have 4 treatments and 7 timepoints. (0, 2, 4, 12, 24, 48, 96 h) We would sample from 5 individual plants that would be pooled for each RNA preparation. This would require 28 chips and this would include extra replication of the 0 time-point control (deemed by many as nessary). Experimental details: All plants were germinated for 7 days under the following conditions: Humax multi-purpose compost, ambient carbon dioxide (370 ppm) and ambient light (250 µmol/m/s), constant temperature of 20°C and a 10 h photoperiod (8 am until 6 pm). After a week the the seedlings were potted up into 104-cell plug trays for a further 2 weeks and then potted up into 10 cm pots and the bottom part of the signalling cuvette system attached (see Lake et al., Nature 10th May 2001 Vol. 411, pp 154). Twenty four, 4 week old plants, then had the top part of the signalling system attached, trapping leaf insertions 5-13. Humidified, ambient air was passed through them at 500 mls/min via an oil-free air compressor. The three target leaves (19-21) were then marked with non-toxic, acrylic paint. After a 24 h period (the plants were sealed into the cuvettes from 10 am until 10am) of adjustment, the experiment was started by harvesting the target leaves from 4 plants and immediately freezing the tissue in liquid nitrogen to give the 0 h sample before RNA extraction. The remaining 20 plants were divided into 4 groups of five and given one of the following treatments: Ambient carbon dioxide/ambient light (Control) (A) Elevated carbon dioxide (750 ppm)/ambient light (E) Ambient carbon dioxide/low light (50 µmol/m/s) (AS) Elevated carbon dioxide/low light (ES) - For the Elevated CO2, elevated CO2 was pumped in using a CT room next door set to same temperature but with a CO2 cylinder inside and the same pump as used in the ambient room to supply the elevated CO2 laden, humidified air into the signalling room using rubber tubing. - Shade treatment consisted of neutral density filter (Cat. 210 0.6ND, Lee Filters) that had a hole cut in the middle to allow the middle developing leaves to grow through. A timecourse of 2, 4, 12, 24, 48 and 96 h were carried out each using a batch of 24 plants. This whole process was repeated with another batch of 24 plants at the same developmental stage to give a 2, 4, 12, 24, 48 and 96 hour sample from each of the four treatments. The whole timecourse was then repeated 4 times. For the mature leaves: We had 8 chips left over so we devised this little experiment to assess the gene changes that were occurring in the enclosed, treated, mature leaves that were signalling the environment to the young developing leaves. Experimenter name = Simon Coupe Experimenter phone = 0114 222 4115 Experimenter fax = 0114 222 0002 Experimenter institute = University of Sheffield Experimenter address = Animal and Plant Sciences Experimenter address = University of Sheffield Experimenter address = Western Bank Experimenter address = Sheffield Experimenter zip/postal_code = S10 2TN Experimenter country = UK Keywords: development_or_differentiation_design; growth_condition_design
Project description:We were awarded a BBSRC grant about a year ago to undertake some affymetrix gene chip profiling of light and CO2 systemic signalling in Arabidopsis. The design of the proposed experiment is given below and the appropriate funding has been provided by the BBSRC. The aim of the project is to identify the temporal profile of those genes that respond to light and CO2 systemic signals in developing leaves. Moreover, as thes two signals have opposing effects on leaf development to ascertain whether they involve similar or parallel signalling pathways. The experiment is to examine the effect of exposing mature leaves to high CO2 or low light or both on the gene expression profile of developing leaves. We already have data for maize that changes in gene expression profile occur within 4h and that there are a variety of temporal responses that differ between individual gene transcripts. We have also demonstrated that Arabidopsis leaf development is altered by these systemmic signals and that lesions in the jasmonate and ethylene signalling pathways block these responses. Our experimental design is shown below: We have 4 treatments and 7 timepoints. (0, 2, 4, 12, 24, 48, 96 h) We would sample from 5 individual plants that would be pooled for each RNA preparation. This would require 28 chips and this would include extra replication of the 0 time-point control (deemed by many as nessary). Experimental details: All plants were germinated for 7 days under the following conditions: Humax multi-purpose compost, ambient carbon dioxide (370 ppm) and ambient light (250 µmol/m/s), constant temperature of 20°C and a 10 h photoperiod (8 am until 6 pm). After a week the the seedlings were potted up into 104-cell plug trays for a further 2 weeks and then potted up into 10 cm pots and the bottom part of the signalling cuvette system attached (see Lake et al., Nature 10th May 2001 Vol. 411, pp 154). Twenty four, 4 week old plants, then had the top part of the signalling system attached, trapping leaf insertions 5-13. Humidified, ambient air was passed through them at 500 mls/min via an oil-free air compressor. The three target leaves (19-21) were then marked with non-toxic, acrylic paint. After a 24 h period (the plants were sealed into the cuvettes from 10 am until 10am) of adjustment, the experiment was started by harvesting the target leaves from 4 plants and immediately freezing the tissue in liquid nitrogen to give the 0 h sample before RNA extraction. The remaining 20 plants were divided into 4 groups of five and given one of the following treatments: Ambient carbon dioxide/ambient light (Control) (A) Elevated carbon dioxide (750 ppm)/ambient light (E) Ambient carbon dioxide/low light (50 µmol/m/s) (AS) Elevated carbon dioxide/low light (ES) - For the Elevated CO2, elevated CO2 was pumped in using a CT room next door set to same temperature but with a CO2 cylinder inside and the same pump as used in the ambient room to supply the elevated CO2 laden, humidified air into the signalling room using rubber tubing. - Shade treatment consisted of neutral density filter (Cat. 210 0.6ND, Lee Filters) that had a hole cut in the middle to allow the middle developing leaves to grow through. A timecourse of 2, 4, 12, 24, 48 and 96 h were carried out each using a batch of 24 plants. This whole process was repeated with another batch of 24 plants at the same developmental stage to give a 2, 4, 12, 24, 48 and 96 hour sample from each of the four treatments. The whole timecourse was then repeated 4 times. For the mature leaves: We had 8 chips left over so we devised this little experiment to assess the gene changes that were occurring in the enclosed, treated, mature leaves that were signalling the environment to the young developing leaves. Experimenter name = Simon Coupe Experimenter phone = 0114 222 4115 Experimenter fax = 0114 222 0002 Experimenter institute = University of Sheffield Experimenter address = Animal and Plant Sciences Experimenter address = University of Sheffield Experimenter address = Western Bank Experimenter address = Sheffield Experimenter zip/postal_code = S10 2TN Experimenter country = UK Keywords: development_or_differentiation_design; growth_condition_design
Project description:To study whether and how soil nitrogen conditions affect the ecological effects of long-term elevated CO2 on microbial community and soil ecoprocess, here we investigated soil microbial community in a grassland ecosystem subjected to ambient CO2 (aCO2, 368 ppm), elevated CO2 (eCO2, 560 ppm), ambient nitrogen deposition (aN) or elevated nitrogen deposition (eN) treatments for a decade. Under the aN condition, a majority of microbial function genes, as measured by GeoChip 4.0, were increased in relative abundance or remained unchanged by eCO2. Under the eN condition, most of functional genes associated with carbon, nitrogen and sulfur cycling, energy processes, organic remediation and stress responses were decreased or remained unchanged by eCO2, while genes associated with antibiotics and metal resistance were increased. The eCO2 effects on fungi and archaea were largely similar under both nitrogen conditions, but differed substantially for bacteria. Coupling of microbial carbon or nitrogen cycling genes, represented by positive percentage and density of gene interaction in association networks, was higher under the aN condition. In accordance, changes of soil CO2 flux, net N mineralization, ammonification and nitrification was higher under the aN condition. Collectively, these results demonstrated that eCO2 effects are contingent on nitrogen conditions, underscoring the difficulty toward predictive modeling of soil ecosystem and ecoprocesses under future climate scenarios and necessitating more detailed studies.
Project description:There are seedling samples (high CO2 exposure of 0h, 2h, 6h, 12h, 1d, 3d, 7d, 14d) with duplicates in two chambers. Arabidopsis WT (Col-0) seeds were plated on Murashige and Skoog plates and placed at 4°C in darkness for at least 2 d to synchronize germination. Plants were grown at 22C under long-day conditions (16-h light and 8-h dark) in two atmospheric CO2 environments: ambient (CO2: 390 μmol molâ??1) or elevated (CO2: 780 μmol molâ??1). Plants were grown in ambient atmospheric CO2 concentration and then exposed to elevated CO2 for 0 h, 2 h, 6 h, 12 h, 1 d, 3 d, 7 d and 14 d. 14-d-old seedlings were sampled at the same time. Treatments for elevated CO2 were carried out two times using two different chambers. Duplicate samples were collected from each chamber experiment. Totally, four biologically replicates were prepared in each condition.
Project description:This work aims to study the effect of the elevated CO2 concentration on the tomato plant response to the toxicity provoked by ammonium nutrition. Tomato plants (Solanum lycopersicum L. cv. Agora Hybrid F1, Vilmorin®) were grown for 4 week with 15 mM of nitrogen, supplied as nitrate or ammonium, at ambient or elevated CO2 conditions (400 ppm or 800 ppm). Transcription profiling by array was carried out in roots for the four growth conditions assayed and gene expression comparisons were done between N sources and CO2 conditions: i) genes differentially expressed in response to the atmospheric CO2 concentration (800 ppm vs 400 ppm CO2) under nitrate or ammonium nutrition; ii) genes differentially expressed in response to the N source (ammonium vs nitrate) under ambient or elevated condition. 3 biological replicates for each growth condition were analysed.CO2).
Project description:The transcript responses of both growing, trifoliate 6 and fully expanded, trifoliate 4 soybean leaves to elevated CO2 was investigated. We also compared the transcriptome of fully expanded vs. developing leaves in both ambient and elevated CO2. Keywords = soybean Keywords = elevated carbon dioxide Keywords = global change Keywords = leaf growth Keywords = plant Keywords: soybean leaf comparisons
Project description:Elevated CO2 (eCO2) has an influence on developing leaf growth of rice (Oryza sativa cv. Nipponbare), specifically lower growth stage than P4 (plastochron number), resulting in decrease in leaf size compared with that in ambient CO2 (aCO2). Since several micro RNAs are associated with the regulation of plant leaf development, in order to clarify which micro RNAs are involved in the decrease of leaf blade size at eCO2, we carried out high-throughput small RNA sequencing analysis and compared the amount of identified miRNAs in developing rice leaf blade grown between aCO2 and eCO2 condition.
Project description:We were awarded a BBSRC grant about a year ago to undertake some affymetrix gene chip profiling of light and CO2 systemic signalling in Arabidopsis. The design of the proposed experiment is given below and the appropriate funding has been provided by the BBSRC. The aim of the project is to identify the temporal profile of those genes that respond to light and CO2 systemic signals in developing leaves. Moreover, as thes two signals have opposing effects on leaf development to ascertain whether they involve similar or parallel signalling pathways. The experiment is to examine the effect of exposing mature leaves to high CO2 or low light or both on the gene expression profile of developing leaves. We already have data for maize that changes in gene expression profile occur within 4h and that there are a variety of temporal responses that differ between individual gene transcripts. We have also demonstrated that Arabidopsis leaf development is altered by these systemmic signals and that lesions in the jasmonate and ethylene signalling pathways block these responses. Our experimental design is shown below: We have 4 treatments and 7 timepoints. (0, 2, 4, 12, 24, 48, 96 h) We would sample from 5 individual plants that would be pooled for each RNA preparation. This would require 28 chips and this would include extra replication of the 0 time-point control (deemed by many as nessary). Experimental details: All plants were germinated for 7 days under the following conditions: Humax multi-purpose compost, ambient carbon dioxide (370 ppm) and ambient light (250 µmol/m/s), constant temperature of 20°C and a 10 h photoperiod (8 am until 6 pm). After a week the the seedlings were potted up into 104-cell plug trays for a further 2 weeks and then potted up into 10 cm pots and the bottom part of the signalling cuvette system attached (see Lake et al., Nature 10th May 2001 Vol. 411, pp 154). Twenty four, 4 week old plants, then had the top part of the signalling system attached, trapping leaf insertions 5-13. Humidified, ambient air was passed through them at 500 mls/min via an oil-free air compressor. The three target leaves (19-21) were then marked with non-toxic, acrylic paint. After a 24 h period (the plants were sealed into the cuvettes from 10 am until 10am) of adjustment, the experiment was started by harvesting the target leaves from 4 plants and immediately freezing the tissue in liquid nitrogen to give the 0 h sample before RNA extraction. The remaining 20 plants were divided into 4 groups of five and given one of the following treatments: Ambient carbon dioxide/ambient light (Control) (A) Elevated carbon dioxide (750 ppm)/ambient light (E) Ambient carbon dioxide/low light (50 µmol/m/s) (AS) Elevated carbon dioxide/low light (ES) - For the Elevated CO2, elevated CO2 was pumped in using a CT room next door set to same temperature but with a CO2 cylinder inside and the same pump as used in the ambient room to supply the elevated CO2 laden, humidified air into the signalling room using rubber tubing. - Shade treatment consisted of neutral density filter (Cat. 210 0.6ND, Lee Filters) that had a hole cut in the middle to allow the middle developing leaves to grow through. A timecourse of 2, 4, 12, 24, 48 and 96 h were carried out each using a batch of 24 plants. This whole process was repeated with another batch of 24 plants at the same developmental stage to give a 2, 4, 12, 24, 48 and 96 hour sample from each of the four treatments. The whole timecourse was then repeated 4 times. For the mature leaves: We had 8 chips left over so we devised this little experiment to assess the gene changes that were occurring in the enclosed, treated, mature leaves that were signalling the environment to the young developing leaves. Experimenter name = Simon Coupe Experimenter phone = 0114 222 4115 Experimenter fax = 0114 222 0002 Experimenter institute = University of Sheffield Experimenter address = Animal and Plant Sciences Experimenter address = University of Sheffield Experimenter address = Western Bank Experimenter address = Sheffield Experimenter zip/postal_code = S10 2TN Experimenter country = UK Keywords: development_or_differentiation_design; growth_condition_design
Project description:To study whether and how soil nitrogen conditions affect the ecological effects of long-term elevated CO2 on microbial community and soil ecoprocess, here we investigated soil microbial community in a grassland ecosystem subjected to ambient CO2 (aCO2, 368 ppm), elevated CO2 (eCO2, 560 ppm), ambient nitrogen deposition (aN) or elevated nitrogen deposition (eN) treatments for a decade. Under the aN condition, a majority of microbial function genes, as measured by GeoChip 4.0, were increased in relative abundance or remained unchanged by eCO2. Under the eN condition, most of functional genes associated with carbon, nitrogen and sulfur cycling, energy processes, organic remediation and stress responses were decreased or remained unchanged by eCO2, while genes associated with antibiotics and metal resistance were increased. The eCO2 effects on fungi and archaea were largely similar under both nitrogen conditions, but differed substantially for bacteria. Coupling of microbial carbon or nitrogen cycling genes, represented by positive percentage and density of gene interaction in association networks, was higher under the aN condition. In accordance, changes of soil CO2 flux, net N mineralization, ammonification and nitrification was higher under the aN condition. Collectively, these results demonstrated that eCO2 effects are contingent on nitrogen conditions, underscoring the difficulty toward predictive modeling of soil ecosystem and ecoprocesses under future climate scenarios and necessitating more detailed studies. Fourty eight samples were collected for four different carbon and nitrogen treatment levels (aCaN,eCaN,aCeN and eCeN) ; Twelve replicates in every elevation
Project description:Elevated CO2 (eCO2) condition has influence on developing leaf growth of rice (Oryza sativa cv. Nipponbare), specifically lower than P4 (plastochron number) stage, resulting in decrease in leaf size compared with that grown at ambient CO2 (aCO2). In this case, decrease in leaf size seems to be one of acclimation process at CO2 replete environment despite of the fact that number of tiller increase during CO2 replete periods; however, it is not yet elucidated which endogenous signal play a precise role in depression of developing leaf growth in those process. In this context, to elucidate precise signal interaction between mature and developing leaf of rice at eCO2 environment, we profiled gene expressions of developing rice leaf (P4) using oligo DNA microarray 4X44K RAP-DB (Agilent Technologies, Santa clara, CA).