ABSTRACT: Transcriptional profiling of Arabidopsis dcp5-1 comparing control WT with 30min dehydration treatment. Goal was to determine the effects of DCP5 on global transcript abundance during dehydration response. Overall design: Two-condition experiment, time0 and 30min dehydration. Biological replicates: 4 WT replicates, 4 dcp5-1 replicates.
INSTRUMENT(S): Agilent-021169 Arabidopsis 4 Oligo Microarray (V4) (Probe Name version)
Project description:Transcriptional profiling of Arabidopsis dcp5-1 comparing control WT with 30min dehydration treatment. Goal was to determine the effects of DCP5 on global transcript abundance during dehydration response. Two-condition experiment, time0 and 30min dehydration. Biological replicates: 4 WT replicates, 4 dcp5-1 replicates.
Project description:To understand the role of KAR-signaling components in water stress response, we have carried out comparative expression analysis of the KAR-receptor kai2-2 mutant and WT plants under dehydration and well-watered (control) conditions. Aligent’s whole Arabidopsis Gene Expression Microarray (G2519F-021169, V4, 4x44K) was used. Overall design: Two-week-old WT and kai2-2 mutant plants were transferred from GM plates to soil and grown for 10 additional day. The aerial parts of 24-d-old plants were detached and exposed to dehydration on KimTowel papers for 0 (well-watered, control), 2 and 4 h. All rosette leaves of independent 24-d-old plants were collected. Total RNA was prepared and used for the microarray hybridization. Three independent biological replicates were used for each plant sample.
Project description:To understand the role of SL-signaling components in water stress response, we have carried out comparative expression analysis of the SL-response max2-3 mutant and WT plants under dehydration and well-watered (control) conditions. Aligent’s whole Arabidopsis Gene Expression Microarray (G2519F-021169, V4, 4x44K) was used. Two-week-old WT and max2-3 mutant plants were transferred from GM plates to soil and grown for 10 additional day. The aerial parts of 24-d-old plants were detached and exposed to dehydration on KimTowel papers for 0 (well-watered, control), 2 and 4 h. All rosette leaves of independent 24-d-old plants were collected. Total RNA was prepared and used for the microarray hybridization. Three independent biological replicates were used for each plant sample.
Project description:Background: Grapevine is a major food crop that is affected by global climate change. Consistent with field studies, dehydration assays of grapevine leaves can reveal valuable information of the plant’s response at physiological, transcript, and protein levels. There are well-known differences in grapevine rootstocks responses to dehydration. We used time-series transcriptomic approaches combined with network analyses to elucidate and identify important physiological processes and network hubs that respond to dehydration in three different grapevine species differing in their drought tolerance. Results: Transcriptomic analyses of the leaves of Cabernet Sauvignon, Riparia Gloire, and Ramsey were evaluated at different times during a 24-h controlled dehydration. ANOVA revealed that 11,000 transcripts changed significantly with respect to the genotype x treatment interaction term and 6,000 transcripts changed significantly according to the genotype x treatment x time interaction term indicating massive differential changes in gene expression over time. Standard analyses determined substantial effects on the transcript abundance of genes involved in the metabolism and signaling of two known plant stress hormones, ABA and ethylene. ABA and ethylene signaling maps were constructed and revealed specific changes in transcript abundance that were associated with the known drought tolerance of the genotypes including genes such as VviABI5, VviABF2, VviACS2, and VviWRKY22. Weighted-gene coexpression network analysis (WGCNA) confirmed these results. In particular, WGCNA identified 30 different modules, some of which had highly enriched gene ontology categories for photosynthesis, phenylpropanoid metabolism, ABA and ethylene signaling. The ABA signaling transcription factors, VviABI5 and VviABF2, were highly connected hubs in two modules, one having overrepresentation in gaseous transport and the other in ethylene signaling. VviABI5 was distinctly correlated with an early response and high expression for the drought tolerant Ramsey and with little response from the drought sensitive Riparia Gloire. These ABA signaling transcription factors were highly connected to VviSNRK1 and other gene hubs associated with sugar, ethylene and ABA signaling. The ABA and ethylene signaling hubs were highly connected, supporting the hypothesis that there is substantial cross-talk between the two hormone pathways. This study identifies solid gene candidates for future investigations of drought tolerance in grapevine. Overall design: Fully mature Cabernet Sauvignon, Riparia Gloire, and Ramsey leaves were dehydrated (Hopper et al., 2014) to assess rapid transcriptomic changes. Briefly, the leaf was excised from the plant and placed into a dehydration box for various time points over a 24-h period. Leaf dehydration occurred in the air above a solution of 333 mM NaCl in a sealed container in a growth chamber. Leaves were removed from the box and immediately frozen in liquid nitrogen. Control samples were taken from the same plant at the corresponding time to account for any circadian effect on transcript abundance. RNA was extracted from three experimental replicates for treatment and control samples at each time point.
Project description:Gene transcript abundances were analyzed with samples taken from hydrated (HD), moderate dehydration (MDH), desiccated (SDH), partially recovered (PRE) and fully recovered (FRE) gametophores of P. patens by using RNA-Seq. Totally, 14686 transcripts were identified as differentially expressed transcripts.The results provided insight for exploring the mechanisms of desiccation tolerance and their evolution. Overall design: Examination of mRNA transcript abundances in hydrated (HD), moderate dehydration (MDH), desiccated (SDH), partially recovered (PRE) and fully recovered (FRE) gametophores, for each treatment, three biological replicates were included.
Project description:Small RNA transcript abundances were analyzed with samples taken from hydrated (HD), moderate dehydration (MDH), desiccated (SDH), partially recovered (PRE) and fully recovered (FRE) gametophores of P. patens by using next generation sequencing. Totally, 147 microRNAs were identified as differentially expressed miRNAs.The results provided insight for exploring the mechanisms of desiccation tolerance and their evolution. Overall design: Examination of miRNA transcript abundances in hydrated (HD), moderate dehydration (MDH), desiccated (SDH), partially recovered (PRE) and fully recovered (FRE) gametophores, for each treatment, two biological replicates were included.
Project description:We report global gene expression profilies of Brassinosteroid related Arabidopsis mutants in response to dehydration and fixed-carbon starvation stresses by RNA-seq Overall design: Arabidopsis plants of listed genotypes were grown for 4 weeks under long day (16 hour light) conditions before being subjected to control, 4 hour dehydration, or 5 day fixed carbon starvation treatments.
Project description:New shoot growth from underground adventitious buds of leafy spurge is critical for survival of this invasive perennial weed after episodes of severe abiotic stress. Because global climate change is expected to increase abiotic stress, such as dehydration, objectives of this study include examining the impact that dehydration stress has on molecular mechanisms associated with vegetative reproduction. Greenhouse plants were exposed to mild- (3-day), intermediate- (7-day), severe- (14-day) and extended- (21-day) dehydration treatments, prior to decapitation of aerial tissue and rehydration of soil to induce new vegetative shoot growth. Compared to well-watered control plants, mild-dehydration accelerated new vegetative shoot growth but intermediate- and severe-dehydration treatments both delayed and reduced shoot growth, and 21-day dehydration treatment inhibited initiation of new vegetative shoots and was considered a lethal treatment. Overall, transcriptome profiles revealed that 2109 genes were differentially-expressed (P<0.05) in crown buds in response to the various dehydration treatments. Sub-network enrichment analyses identified central hubs of over-represented genes involved in processes such as hormone responses and signaling (e.g., ABA, auxin, ethylene, GA, and JA), response to abiotic stress (DREB1A/2A) and light (PIF3), phosphorylation (CLV1, MPK3/4/6, SOS2), gene silencing (miRNA156/172a), circadian regulation (CRY2, LHY, PHYA/B), and flowering (AGL8/20, AP2, FLC). Further, results from this and previous studies highlight HY5, MAF3, MYB-like/RVE1 and RD22 as molecular markers for endodormancy in crown buds of leafy spurge. Early response to dehydration also highlighted involvement of upstream ethylene and jasmonate signaling, whereas longer-term dehydration impacted ABA signaling. The identification of conserved ABRE- and MYC-consensus, cis-acting elements in the promoter of a leafy spurge gene similar to Arabidopsis MYB-like/RVE1 (AT5G17300) implicates a potential role for ABA signaling in its dehydration-induced expression. Response of these molecular mechanisms to dehydration-stress provides insights on the ability of invasive perennial weeds to adapt and survive under harsh environments, which provide new insights for addressing future management practices. Changes in transcript abundance for underground adventitious buds of leafy spurge which were exposed various levels of dehydration stress (Day-3, -7, -14, -16, -21) are analysed relative to controls (Day-0).
Project description:Arabidopsis plants that have experienced stress from water withdrawal show an improved ability to tolerate subsequent exposures as a ‘memory’ from the previous stress. This physiological stress memory is associated with ‘transcriptional memory’ illustrated by a subset of dehydrations stress responding genes that produce significantly different transcript amounts during repeated dehydration stresses relative to their response in the first. Here we report the genome-wide representation of dehydration stress transcriptional memory genes in A. thaliana. We identify four novel transcription patterns in response to repeated dehydration stress treatments. The nature of the proteins encoded by genes from each type of memory-response pattern is analyzed and the consequences of the genes’ memory behavior are considered in the context of possible biological relevance. The memory behavior of genes co-regulated by the dehydration/ABA and other abiotic stress and hormone responding pathways suggested that the crosstalk at the transcriptional level between them was affected as well. The intensity and the nature of specific biochemical, membrane, chloroplast, and stress response-related interactions during multiple exposures to dehydration stress are different from the responses to a single dehydration stress. The results reveal additional, hitherto unknown, levels of complexity of the plants’ transcriptional behavior when adjusting and adapting to recurring water deficits. For each condition (water, S1, and S3) the transcriptome was sequenced for two replicates. The watered condition is considered the control.
Project description:To understand the Abscisic Acid (ABA) signaling in response to dehydration stress, we performed analysis of gene expression using Arabidopsis wild-type plants and the nced3-2 mutant under dehydration stress. The nced3-2 mutant is an Arabidopsis T-DNA tagged knock-out mutant of the NCED3 gene, which has an essential role in dehydration-inducible ABA biosynthesis. Arabidopsis plants were grown in in soil (verdenite 40 mmΦ, Verde Co., Ltd., Kanagawa, Japan) in a cell strainer (Falcon, 40 μm; Corning Inc., NY, USA). Plants were grown at 22°C for 3 weeks under illumination (40–60 μmol m-2s-1; 16 h light/8 h darkness). Three-week-old plants were exposed to dehydration stress by being denied water for 6, 24, 48, or 72 h.