Project description:Polyethilenimine (PEI) functionalized single walled carbon nanotubes (SWNTs) and carbon-based nanomaterials enable delivery of DNA and RNA in plants. Given the broad-scale use of PEI-functionalized nanomaterials in plants, we sought to investigate the reaction of plant tissues to treatment with PEI-SWNTs and pristine SWNTs. To this end, we infiltrated Arabidopsis thaliana leaves with pristine single walled carbon nanotubes used in RNA silencing applications (SWNTs) and polyethyleneimine-functionalized SWNTs used for plasmid DNA delivery (PEI-SWNTs). We used Arabidopsis as it is a well characterized model plant, for which genomic and detailed gene function information is readily available. To minimize the effects caused by the introduction of exogenous nucleic acids, in SWNT preparations we used single stranded RNA targeting Green Fluorescent Protein (GFP) with no target sequence in the Arabidopsis genome, and a plasmid that expresses GFP in PEI-SWNT preparations. For our experiments herein, we used ~25-50 fold higher concentrations of SWNTs and PEI-SWNTs compared to standard concentrations used in biomolecule delivery assays. Water-infiltrated plant leaves served as a negative control to distinguish between the SWNT-specific response and the response to the infiltration process itself. We performed RNA sequencing (RNA-seq) with RNA extracted from leaves two days after infiltration to identify changes in the leaf transcriptomic profile in response to the three treatments, compared to non-infiltrated leaves.
Project description:Basal leaf removal has been widely practiced to increase cluster sunlight exposure, control berry rot and eventually improve grape berry quality. Early leaf removal performed in cool region appeared to benefit the accumulation of norisoprenoids and monoterpenes. However, effects of this canopy management and leaf moving on volatile profiles and transcriptome of grape berry in warm region characterized with strong sunshine and arid climate were paid less attention. To cope with multiple possible effects caused by global warming, researches in warm region are urgently needed to provide a guide for adjusting the sunlight exposure treatment to adapt the climate change.In Manas county of Xinjiang province, sunlight exposure treatments performed in this study resulted in decreased β-carotene and lutein, which mainly responsible for the reduced norisoprenoids in ripening grapes. Substrate competition due to the up-regulation of VvTPS-a and VvNCED3 may contribute to the decreased concentration of monoterpenes in exposed berries. The notable increased C6 alcohols in the grape berries treated by leaf removal applied at veraison (LR-V), was mainly attribute to the enhanced substrate availability of linoleic acid and linolenic acid together with the higher expression of VvLOXO and VvADH1. Free C6 aldehyde was reduced by half leaf removal applied at veraison (HLR-V), leaf moving at veraison (LM-V) and leaf removal applied at berry pepper-corn size (LR-PS). Moreover, transcriptomic analysis indicated that both LR-V and LM-V treated grape berry implemented multiple stress-mitigation strategies to acclimate the improved sunlight exposure. Interestingly, photosynthesis-related genes in grape berry were primarily up-regulated by LR-V as a consequence of carbohydrate source removal rather than improved sunlight exposure. Weighted gene co-expression network analysis (WGCNA) suggested that the genes encoding malate synthase and 3-oxoacyl-(acyl carrier protein) reductase show a significant correlation with the accumulation of C6 alcohol.
Project description:gene expression profiling in different zones along the gradient of the growing maize leaf balde aover a time course of dirunal cycle and carbon starvation by extension of the night Plants assimilate carbon in their photosynthetic tissues in the light. However, carbon is required during the night, and in non-photosynthetic organs. It is therefore essential that plants manage their carbon resources spatially and temporally and coordinate growth with carbon availability. In growing maize (Zea mays) leaf blades a defined developmental gradient facilitates analyses in the cell division, elongation and mature zones. We investigated the responses of the metabolome and transcriptome and polysome loading, as a qualitative proxy for protein synthesis, at dusk, dawn and 6, 14 and 24 hours into an extended night, and tracked whole leaf elongation over this time course. Starch and sugars are depleted by dawn in the mature zone, but only after an extension of the night in the elongation and division zones. Sucrose recovers partially between 14 and 24 h into the extended night in the growth zones but not the mature zone. The global metabolome and transcriptome track these zone-specific changes in sucrose. Leaf elongation and polysome loading in the growth zones also remain high at dawn, decrease between 6 and 14 h into the extended night and then partially recover indicating that growth processes are determined by local carbon status. The level of sucrose-signaling metabolite trehalose-6-phosphate, and the trehalose-6-phosphate:sucrose ratio are much higher in growth than mature zones at dusk and dawn but fall in the extended night. Candidate genes were identified by searching for transcripts that show characteristic temporal response patterns or contrasting responses to carbon starvation in growth and mature zones.
Project description:Heat stress is one of the primary abiotic stresses that limit crop production . Grape is a popular cultivated fruit with high economic value throughout the world, and whose growth and development is often influenced by high temperature. Alternative splicing (AS) is a widespread mechanism increasing transcriptome complexity and proteome diversity. We conducted high temperature treatments (35oC, 40oC and 45oC) on grapevines (Vitis vinifera), and assessed proteomic and transcriptomic (especially AS)changes in leaves. We found that nearly 70% of the genes were alternatively spliced under high temperature. Intron retention (IR), exon skipping (ES) and alternative donor/acceptor sites were markedly induced under different high temperatures. IR was the most abundant up- and down-regulated AS event; moreover, IR events at 40 and 45oC were far higher than those at 35oC. These results indicated AS, especially IR, is an important posttranscriptional regulatory during grape leaf responses to high temperature. Proteomic analysis showed that protein levels of the RNA binding proteins SR45, SR30, and SR34, and the nuclear ribonucleic protein U1A in grape leaves gradually rose as ambient temperature increased. The results also revealed why AS events occurred more frequently under high temperature in grape leaves. After integrating transcriptomic and proteomic data, we found that HSPs and some important transcript factors such as MBF1c and HSFA2 were mainly involved in heat tolerance in grape through up-regulating transcriptional and translational levels, and were especially modulated by AS. The results provide the first simultaneous evidence for grape leaf responses to high temperature at transcriptional, posttranscriptional and translational levels.
Project description:gene expression profiling in different zones along the gradient of the growing maize leaf balde aover a time course of dirunal cycle and carbon starvation by extension of the night Plants assimilate carbon in their photosynthetic tissues in the light. However, carbon is required during the night, and in non-photosynthetic organs. It is therefore essential that plants manage their carbon resources spatially and temporally and coordinate growth with carbon availability. In growing maize (Zea mays) leaf blades a defined developmental gradient facilitates analyses in the cell division, elongation and mature zones. We investigated the responses of the metabolome and transcriptome and polysome loading, as a qualitative proxy for protein synthesis, at dusk, dawn and 6, 14 and 24 hours into an extended night, and tracked whole leaf elongation over this time course. Starch and sugars are depleted by dawn in the mature zone, but only after an extension of the night in the elongation and division zones. Sucrose recovers partially between 14 and 24 h into the extended night in the growth zones but not the mature zone. The global metabolome and transcriptome track these zone-specific changes in sucrose. Leaf elongation and polysome loading in the growth zones also remain high at dawn, decrease between 6 and 14 h into the extended night and then partially recover indicating that growth processes are determined by local carbon status. The level of sucrose-signaling metabolite trehalose-6-phosphate, and the trehalose-6-phosphate:sucrose ratio are much higher in growth than mature zones at dusk and dawn but fall in the extended night. Candidate genes were identified by searching for transcripts that show characteristic temporal response patterns or contrasting responses to carbon starvation in growth and mature zones. 3 repliucates per time point and leaf region, each pooled form 5 indiviual plants
Project description:The aim of our study is to investigate the effects of carbon ion and photon irradiation on A549 tumor cells and analyse how these effects are altered by PML knockdown. Therefore we created PML knockdown A549 cells (shPML) and irradiated them with either 2Gy carbon ion or 6Gy Photon (bioequivalent doses). 4 days after irradiation microarray analysis was performed. All experiments were performed in 3 biological replicates and control groups were transduced with an empty vector.
Project description:In order to screen for molecules contributing to the effects of FGF1/CPP-C on radiation-induced intestinal damage, the total RNA expression profile of the irradiated jejunum was obtained using a DNA microarray analysis after Carbon-ion irradiation with the FGF1/CPP-C pretreatment.
Project description:One-carbon metabolism is an essential branch of cellular metabolism that intersects with epigenetic regulation. Here, we show formaldehyde, a one-carbon unit derived from both endogenous sources and environmental exposure, regulates one-carbon metabolism by inhibiting the biosynthesis of S-adenosylmethionine (SAM), the major methyl donor in cells. Formaldehyde reacts with privileged, hyperreactive cysteine sites in the proteome, including Cys120 in S-adenosylmethionine synthase isoform type-1 (MAT1A). Formaldehyde exposure inhibited MAT1A activity and decreased SAM production with MAT-isoform specificity. A genetic mouse model of chronic formaldehyde overload showed a decrease in SAM and in methylation on selected histones and genes. Epigenetic and transcriptional regulation of Mat1a and related genes function as compensatory mechanisms for formaldehyde-dependent SAM depletion, revealing a biochemical feedback cycle between formaldehyde and SAM one-carbon units.