Dataset Information

Time-resolved metabolomics and proteomics of Brassica napus guard cells under low CO2 concentration

ABSTRACT: In order to discover new nodes and edges for low CO2-induced stomata closure, we applied hyphenated mass spectrometry (MS)-based metabolomics and proteomics approaches to analyze short-term low CO2 responses in B. napus (canola) guard cells. A total of 411 metabolites were quantified. We observed decreased trends of primary metabolites biosynthesis (e.g., most common amino acids, nucleotides, and sugars), and changes in the levels of osmoregulators such as sucrose, malate, and mannitol showed increased trends at both early and late time points. Unlike under elevated CO2 levels, JA biosynthesis was not altered. Instead, phytohormones that induces stomatal opening, including cytokinins, auxins, GA4, as well as melatonin increased at early time points. This study also highlights the utility of single cell-type metabolomics in discovering and testing new nodes and edges in cellular signaling and metabolic networks.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Brassica Napus (rape)

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Ning Zhu

LAB HEAD: Ning Zhu

PROVIDER: PXD005787 | Pride | 2018-10-26

REPOSITORIES: Pride

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Publications

Metabolomics and Proteomics of Brassica napus Guard Cells in Response to Low CO2.

Geng Sisi S Yu Bing B Zhu Ning N Dufresne Craig C Chen Sixue S

Frontiers in molecular biosciences 20170725

Stomatal guard cell response to various stimuli is an important process that balances plant carbon dioxide (CO2) uptake and water transpiration. Elevated CO2 induces stomatal closure, while low CO2 promotes stomatal opening. The signaling process of elevated CO2 induced stomatal closure has been extensively studied in recent years. However, the mechanism of low CO2 induced stomatal opening is not fully understood. Here we report metabolomic ...[more]

PMID: 28791296

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Project description:• Reference to published study making use of this data: • Cseke LJ, Tsai C-J, Rogers A, Nelsen MP, White HL, Karnosky DF, Podila GK. (2009) Transcriptomic comparison in the leaves of two aspen genotypes having similar carbon assimilation rates but different allocation patterns under elevated CO2. New Phytologist. submitted. • This study compared the leaf transcription profiles, physiological characteristics, and primary metabolites of two Populus tremuloides genotypes (clones 216 and 271) known to differ in their responses to long-term elevated [CO2] (e[CO2]) at the Aspen FACE site near Rhinelander, WI. • Physiological responses of these clones are similar in photosynthesis, stomatal conductance, and leaf area index under e[CO2] yet very different in growth enhancement (0-10% in clone 216; 40-50% in clone 271). While few genes responded to long-term exposure to e[CO2], the transcriptional activity of leaf e[CO2]-responsive genes was distinctly different between the clones, differentially impacting multiple pathways during both early and late growing seasons. • Analysis of transcript abundance and carbon/nitrogen biochemistry suggests that the CO2-responsive clone (271) partitions C into pathways associated with active defense/response to stress, carbohydrate/starch biosynthesis and subsequent growth. The CO2-unresponsive clone (216) partitions C into pathways associated with passive defense (e.g. lignin, phenylpropanoid) and cell wall thickening. • This study indicates that there is significant variation in expression patterns between different tree genotypes in response to long-term exposure to e[CO2]. Consequently, future efforts to improve productivity or other advantageous traits for carbon sequestration should include an examination of genetic variability in CO2 responsiveness. Keywords: Tree genotype comparison under elevated [CO2] 24 two-channel arrays, directly comparing RNA from trees grown in the ambient (control) [CO2] to RNA derived from the same genotype grown under e[CO2]. For each of 4 experimental conditions (clone 216 early season; clone 271 early season; clone 216 late season; clone 271 late season), three independent biological replicates derived from trees grown in three independent replicate FACE rings were used. In addition, each clone and time point included dye swap reciprocal two-color experiments for each biological replicate. Thus, six data points per cDNA are included (three biological replicates with two technical replicates each).

Project description:Over recent decades the progression of natural autumnal senescence has been increasingly delayed across wide areas of the Northern Hemisphere and is thought to be caused by rising temperature. Recently this delay has been shown to be partly attributable to rising atmospheric carbon dioxide concentrations [CO2]. Here, for the first time, we present a possible mechanistic explanation for this phenomenon. Using a plantation of Populus x euramericana grown at elevated [CO2] (e[CO2]) using Free-Air CO2 Enrichment (FACE) technology, we investigated the molecular and biochemical basis underlying this response. Using a poplar cDNA microarray where late growing season and senescing leaves from ambient [CO2] (a[CO2]) and e[CO2] were compared, revealed unique increases and decreases in transcript abundance in the e[CO2] treatment. Growth at e[CO2]caused the greatest increase in the abundance of transcripts catalysing steps in the anthocyanin biosynthetic pathway and an increase in leaf anthocyanin content. Leaf sucrose and starch content were also increased during senescence in e[CO2] and associated with a higher abundance of transcripts in the sucrose and starch biosynthetic pathways. We propose that in e[CO2], autumnal photosynthesis and sugar accumulation results in changes in genes expression that include further investment in secondary carbon metabolism leading to anthocyanin production. This enables prolonged leaf longevity during natural autumnal senescence through increased availability of carbon and improved stress tolerance while in contrast this may also have a negative effect on the control of dormancy. Samples GSM398292..GSM398314 (late senescence samples only): Six replicate trees were labelled in the unfertilised sub-plots of plots 1-4 in the POP/EUROFACE site (http://www.unitus.it/euroface/) and the 10th leaf down from the closed apical bud of the dominant branch were labelled. Leaves were sampled to liquid nitrogen (18th October 2004) and stored at -80 oC. Extracted RNA was tested for quality and quantity and of that passing the test equal quantities of RNA from each replicate were taken and each sample from the control plots (plots 2 and 3) were randomly paired with a sample from and elevated CO2 plot (plots 1 and 4). From the resulting 9 dual channel hybridisations 6 had the control sample in the Cy3 channel and 3 had the elevated CO2 sample in the Cy3 channel. The 6 samples with Cydye orientation as Cy3=control (ambient CO2) and Cy5 = treated (elevated CO2) are as follows: GSM398296, GSM398301, GSM398303, GSM398304, GSM398305 and GSM398306. The 3 samples with dye swap i.e Cy3 = treated (elevated CO2) are as follows and Cy5 = control (ambient CO2) are as follows: GSM398292, GSM398313 and GSM398314. Samples GSM398426..GSM398474: Four replicate leaves per sub-plot were sampled and flash frozen into liquid nitrogen, on 31 August 2004, and prior to visible senescence but around the time of bud-set as given by Calfapietra et al., (2003) [1]. The tenth leaf down the dominant stem (which represented the first fully expanded mature leaf of maximum size) from a reference leaf of 30 mm in length (leaf one) were selected. On the 14th September twenty trees from within each experimental plot were taken. The tenth leaf down the dominant stem (which represented the first fully expanded mature leaf of maximum size) from a reference leaf of 30 mm in length (leaf one) were labelled on the main-stem above the petiole. All labelled leaves were a uniform height (approx 0.3 m) from the canopy top across all treatments and not shaded by the canopy. Four replicate leaves from within each sub-plot were sampled during each harvest (except on 18th October when six replicate leaves were sampled). Each sampling time was restricted to two hours within the same day and across sampling day’s uniform cloud free skies were chosen. September samplings took place between 15.00 and 17.00 when climatic variables were generally consistent; later in the season time of sampling was adjusted to an earlier two hour period after mid-day when climatic variables were generally consistent. A chlorotic canopy was evident in plots five and six, as described by Liberloo et al. (2007) therefore any subsequent analysis was for plots one to four only. Six replicate labelled leaves were sampled (as above) from each sub-plot on 18th October 2004

Project description:Genome-wide analysis of gene expression has documented differential gene expression between closely related species in plants or animals and nonadditive gene expression in hybrids and allopolyploids compared to the parents. In Arabidopsis, 15-43% of genes are expressed differently between the related species, A. thaliana and A. arenosa, and the majority of them are nonadditvely expressed (differently from mid-parent value) in allotetraploids relative to the parents. The gene expression variation is associated with transcriptional and posttranscriptional mechanisms. Although transcriptional regulation by epigenetic mechanisms plays a role in nonadditive gene expression, the effects of posttranscriptional regulation on nonadditive gene expression is largely unknown. Here we reported a comprehensive genome-wide analysis of mRNA decay in Arabidopsis allotetraploids using oligo gene microarrays. Among ~26,000 annotated genes on the arrays, over 1% of genes showed a rapid decay rate with an estimated half-life of less than 60 min, and these genes are known as genes with unstable transcripts (GUTs). Interestingly, 30% of GUTs matched the nonadditively expressed genes, and their expression levels were significantly negatively correlated. These nonadditively expressed GUTs are highly enriched with a 2-8-fold increase compared to the proportion of all genes in the Gene Ontology (GO) classifications in response to stress, abiotic and biotic stimulus, signal transduction, and transcription. The data are consistent with the enrichment of nonadditvely-expressed genes in those categories in allopolyploids of Arabidopsis, wheat and cotton. Interestingly, the GUTs include some key transcription factors including circadian clock regulators, suggesting a mechanism for rapid response to light signalling and clock regulation in allotetraploids. These data collectively suggest an important role of mRNA stability in nonadditive expression of homoeologous genes in allopolyploids. A total of 8 slides were used for comparison (0 min vs. 120 min), including two biological and four technical replications. Each biological replication consists of two technical replications (four slides) that were hybridized with reverse-labeled probes.

Dataset Information

Time-resolved metabolomics and proteomics of Brassica napus guard cells under low CO2 concentration

Publications

Metabolomics and Proteomics of <i>Brassica napus</i> Guard Cells in Response to Low CO<sub>2</sub>.

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