Project description:The most effective method of micro-propagation for clonal replicates for switchgrass seed production is from the axillary buds of the lower nodes. We hypothesized that buds and nodes from low and high tillering lines from an inbred population will aid in identifying genes important for the different tillering habits of these plants. Since tiller number is directly correlated with biomass yields, identifying genes associated with high tillering trait will provide valuable biomarkers for marker-assisted selection and for QTL mapping of tillering trait in switchgrass. First-generation switchgrass inbred lines derived from selfing a parental genotype M-^QNL94 LYE 16x13M-^R, which was selected from the Oklahoma State University northern lowland breeding population in 2007 was used in this study. The NL94/298 inbred line produced fewer tillers than NL94/145. Stems of the high tillering line had 2-3 phytomers, while the low tillering line had 1-2 phytomers. Eight other inbred lines from this population that showed similar phenotypes as NL94/145 or NL94/298 and similar genotypes based on 380 genomic SSR marker profiles were selected for this study. Individual stems from these nine lines each for high and low tillering habit were harvested from field growing plants in the Agronomy field plots, Stillwater. Vegetative buds from first phytomer and node regions from second phytomer were cut out using a sharp scalpel under a dissection scope and immediately frozen in liquid nitrogen for transcriptomics analysis.
Project description:Small RNA libraries were constructed, sequenced, and analyzed with our Sequence Homology Pipeline for miRNA discovery (Jeong et al. 2013) to identify 59 unique conserved miRNA sequences from 199 precursors in switchgrass.
Project description:Sustainable production of switchgrass (Panicum virgatum) as a bioenergy crop hinges in part on efficient use of soil macronutrients, especially nitrogen (N). This study investigated the physiological, metabolic and transcriptomic responses of switchgrass to N limitation. Moderate N limitation marked a tipping point for large changes in plant growth, root-to-shoot ratio, root system architecture and total nitrogen content. Integration of transcriptomic and metabolic data revealed that N limitation reduced switchgrass photosynthetic capacity and carbon(C)-fixation activities. Switchgrass balanced C-fixation with N-assimilation, transport and recycling of N compounds by rerouting C-flux from glycolysis, the oxidative branch of the pentose phosphate pathway (OPPP) and from the tricarboxylic acid (TCA) cycle in an organ specific manner. The energy and reduction power so generated, and C-skeletons appear to be directed towards N uptake, biosynthesis of energy storage compounds with high C/N ratio such as sucrose, non-N-containing lipids, and various branches of secondary metabolism.
Project description:Purpose: Increasing biomass yield and quality of feedstock have been a recent interest in switchgrass research. Despite the economic importance of switchgrass, increasing temperature and water deficit are limiting factors to the cultivation of bioenergy crops in the semi-arid areas. The effect of individual drought or heat stress has been studied separately in switchgrass. However, there is relatively limited or no report on the molecular basis of combined abiotic stress tolerance in switchgrass particularly the combination of drought and heat stress. We used RNA-Seq approaches to elucidate the transcriptome changes of switchgrass in response to drought and high temperatures simultaneously. Method: We conducted solely drought treatment in switchgrass plant Alamo AP13 by withholding water after 45 days of growing. For the combination of drought and heat effect, heat treatment (35 °C/25 °C day/night) was imposed after 72 h of the initiation of drought. Samples were collected at 0 h, 72 h, 96 h, 120 h, 144 h, and 168 h after treatment imposition, total RNA was extracted, and RNA-Seq conducted. Results:Out of total 32,190 genes, we identified 3,912, as DT responsive genes, 2,339 and 4,635 as , heat (HT) and drought and heat (DTHT) responsive genes, respectively. There were 209, 106, and 220 transcription factors (TFs) differentially expressed under DT, HT and DTHT respectively Conclusion: Through RNA-Seq analysis, we have identified unique and overlapping genes in response to DT and combined DTHT stress in switchgrass. The combination of DT and HT stress may affect the photosynthetic machinery and phenylpropanoid pathway of switchgrass which negatively impacts lignin synthesis and biomass production of switchgrass. The biological function of genes identified particularly in response to DTHT stress could further be confirmed by techniques such as single point mutation or RNAi.
Project description:The combined heat and drought stress influence the plant growth and development. Switchgrass is an economically important crop due to the availability of high biomass with little water and nutrient requirements. Earlier reports suggested that switchgrass growth and yield highly influenced by heat and drought. The mechanism behind heat and drought stress is not fully understood in switchgrass. This study has undertaken to analyze the epigenetic modification using ChIP-Seq analysis with the activation histone mark H3K4me3. Conclusion: Our study provides the first epigenomic analysis of heat and drought response in switchgrass. This comprehensive resource will provide other epigenomic regulated information in non-model plant species.
Project description:In this study we were interested in identifying the gene networks that are responsive to chronic heat treatment in a switchgrass cultivar that is widely grown in the south-western USA and where 38C day temperatures have been reported for more than 100 days in the recent years. Switchgrass Alamo plants were subjected to chronic heat stress for 50 days (38 C/30C; day/night) or maintained under optimal conditions (28C/20C). Leaves were collected at the end of the heat regime for transcriptome analysis.
Project description:Switchgrass (Panicum virgatum L.) has been subject to breeding to improve its yield and composition for bioenergy, but improving its tolerance to environmental variability is just beginning. Different ploidy populations act as somewhat distinct gene pools which can only be bridged through whole genome reduction or duplication events. In order to document potential wide-ranging effects of whole genome duplication, we examined the effects of water stress on growth, physiology, and gene expression in individual tetraploid clones of the switchgrass cultivar ‘Liberty’ as well as neo-octoploid lines derived from it. The neo-octoploids behaved similarly to Liberty under water-stress and recovery conditions. Growth rates, photosynthesis, gas exchange, node numbers, and height were reduced in plants under water stress while proline levels were increased. A total of 6134 differentially expressed genes (8% of the annotated genes with detectable expression in crown tissue) were detected under water deficit stress, while 3310 differentially expressed genes were detected in crown tissue after 1 week of recovery from water deficit stress relative to well-watered treatments. Only a small number of genes were identified as being differentially expressed between 4x Liberty and its 8x derivatives.
Project description:Switchgrass is currently one of the plant species being investigated as a potential biofuels crop. Genetic manipulation can be used to optimize and improve switchgrass to have more properties better suited for biofuels production. We have conducted a proteome with a focus on the endomembrane system of switchgrass so candidate proteins will be available that can be used for genetic manipulation. This proteomic resource will supplement the developing switchgrass genome sequencing project.