Project description:Transcriptome changes were investigated for Euphorbia esula (leafy spurge) seeds with a focus on the effect of constant and diurnal fluctuating temperature on dormancy and germination. Leafy spurge seeds do not germinate when incubated for 21 days at 20°C constant temperatures, but nearly 30% germinate after 21 days under fluctuating temperatures 20:30°C (16:8 h). Incubation at 20°C for 21 followed by 20:30°C resulted in approximately 63% germination in about 10 days. A cDNA microarray representing approximately 22,000 unique sequences was used to profile transcriptome changes.
Project description:Bud dormancy in perennials in boreal and temperate ecosystems is crucial for survival in harsh winter. Dormancy is released by prolonged exposure to low temperatures and is followed by reactive growth in the spring. Lysine acetylation (Kac) is one of the major post-translational modifications (PTMs) involved in plant response to environment signals. However, little information is available on the effects of Kac modification on bud dormancy release. Here, we report the dynamics of lysine acetylome in hybrid poplar (Populus tremula x alba) dormant buds. A total number of 7,594 acetyl sites from 3,281 acetyl proteins were identified, representing the largest to date dataset of lysine acetylome in plants. Of them, 229 proteins were differentially acetylated during bud dormancy release and were involved mainly in the primary metabolism. Site-directed mutagenesis enzymatic assays showed that acetylation strongly modified the activities of two key enzymes of primary metabolism, pyruvate dehydrogenase (PDH) and isocitrate dehydrogenase (IDH). We thus propose that Kac of enzymes could be an important strategy for reconfiguration of metabolic processes during bud dormancy release. In all, our results reveal the importance of Kac in bud dormancy release and give a new perspective to understand the molecular mechanisms of tree’s seasonal growth.
Project description:Transcriptome changes were investigated for Euphorbia esula (leafy spurge) seeds with a focus on the effect of constant and diurnal fluctuating temperature on dormancy and germination. Leafy spurge seeds do not germinate when incubated for 21 days at 20°C constant temperatures, but nearly 30% germinate after 21 days under fluctuating temperatures 20:30°C (16:8 h). Incubation at 20°C for 21 followed by 20:30°C resulted in approximately 63% germination in about 10 days. A cDNA microarray representing approximately 22,000 unique sequences was used to profile transcriptome changes. Labeled cDNA was prepared from total RNA using the Alexa Fluor cDNA labeling kit (Invitrogen, Carlsbad, CA) according to manufacture's protocols. Labeled cDNAs were hybridized to a custom made 23 K element microarray that contained 19,808 unigenes from the leafy spurge EST database and an additional 4,129 unigenes from a cassava EST database. A rolling circle dye swap hybridization scheme was used to compare gene expression between samples. There were three biological and two technical replications for each treatment. Microarray hybridization was visualized using a GenPix 4000B scanner (Axon Instruments/Molecular Devices Corp., Sunnyvale, CA) and spot intensities and background was quantified using GenPix Pro software. Hybridization intensities were log2 transformed, and arrays were centered and normalized against each other.