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Marginal land suitability for switchgrass, Miscanthus and hybrid poplar in the Upper Mississippi River Basin (UMRB)
Not provided.
Novel transcription factors PvBMY1 and PvBMY3 increase biomass yield in greenhouse-grown switchgrass (Panicum virgatum L.)
Not Available
Impacts on soil nitrogen availability of converting managed pine plantation into switchgrass monoculture for bioenergy
Not Available
Carbohydrate and Lignin are Simultaneously Solubilized from Unpretreated Switchgrass by Microbial Action at High Temperature
Abstract not provided
Characterization of organosolv switchgrass lignin by using high performance liquid chromatography/high resolution tandem mass spectrometry using hydroxide-doped negative-ion mode electrospray ionization
Method for identification of compounds in organosolv lignin.
Evaluation of process severity on the chemical composition of organosolv switchgrass lignins by using mass spectrometry
Organosolv treatment is an environmentally friendly fractionation method that affords a lignin stream with high purity.
Conversion of ammonia-pretreated switchgrass to biofuel precursors by bacterial-fungal consortia under solid-state and submerged-state cultivation
Not provided.
Enhanced Characteristics of Genetically Modified Switchgrass (Panicum virgatum L.) for High Biofuel Production
Abstract not provided
A Genome-Wide Survey of Switchgrass Genome Structure and Organization
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Identifying Differences in Abiotic Stress Gene Networks between Lowland and Upland Ecotypes of Switchgrass (DE-SC0008338)
Switchgrass (Panicum virgatum) is a warm-season C4 grass that is a target lignocellulosic biofuel species for use in the United States due to its local adaption capabilities and high biomass accumulation. Two ecotypes of switchgrass have been described. Members of the lowland ecotype are taller, have narrower leaf blades and generate more biomass compared to individuals from the upland ecotype. Additionally, lowland plants are generally found in the southern United States while upland switchgrass is more typically present in the northern United States. These differences are important as it is envisioned that switchgrass for biofuel production will typically be grown on marginal lands in the northern United States to supplement and diversify farmers' traditional crop incomes. While lowland switchgrass is more productive, it has poor winter survivability in northern latitudes where upland switchgrass is expected to be grown for biofuel use. Abiotic stresses likely to be encountered by switchgrass include drought and salinity. Despite initially being described as preferring wetter environments, members of the lowland ecotype have been characterized as being more drought tolerant than plants of the upland ecotype. Nonetheless, direct trials have indicated that variation for drought tolerance exists in both ecotypes, but prior to this project, only a relatively small number of switchgrass lines had been tested for drought responses. Similarly, switchgrass cultivars have not been widely tested for salt tolerance, but a few studies have shown that even mild salt stress can inhibit growth. The effects of drought and salt stress on plant growth are complex. Both drought and salinity affect the osmotic potential of plant cells and negatively affect plant growth due to reduced water potential and reduced photosynthesis that results from lower stomatal conductance of CO 2 . Plants respond to drought and salt stress by activating genes that directly attempt to reduce the stress (e.g., transmembrane pumps that partition Na + ) and mitigate the effects of the stress (e.g., synthesis of osmoprotectant metabolites and stress-related signaling compounds). Prior to the start of this project, no gene expression analysis had been performed on switchgrass under conditions of drought or salt stress, and therefore, relevant gene networks responding to drought and salt stress were unknown in switchgrass. In this project, we performed drought, salt and alkali-salt screens on 49 switchgrass cultivars (Liu et al 2014; Liu et al 2015; Hu et al 2015; Kim et al 2016). These experiments demonstrated that a wide range of variation exists within switchgrass for drought, salt and alkali-salt tolerance and that, while the lowland ecotype of switchgrass is often considered more tolerant of abiotic stresses, there are some upland switchgrass lines that are also very tolerant of drought, salt and alkali-salt stress. We also conducted drought and salt time course experiments with Alamo and Dacotah. We have identified modules of coexpressed genes that differentiate Alamo and Dacotah drought responses. We are continuing to analyze these results and plan to submit manuscripts describing this work in early 2017. In an effort to show how drought- and salt-related gene modules could be dissected, we generated transgenic switchgrass overexpressing either PvGTγ-1 or ZmDREB2. Increased expression of PvGTγ-1 does confer increased salt tolerance, and we were able to identify genes that are induced and suppressed by PvGTγ-1. Overexpression of ZmDREB2 increases drought tolerance in switchgrass. Analysis of the PvGTγ-1 and ZmDREB2 overexpression work is ongoing, and we plan to prepare manuscripts about these experiments for submission in early 2017.
Association Mapping of Cell Wall Synthesis Regulatory Genes and Cell Wall Quality in Switchgrass
Inefficient conversion of biomass to biofuels is one of the main barriers for biofuel production from such materials. Approximately half of polysaccharides in biomass remain unused by typical biochemical conversion methods. Conversion efficiency is influenced by the composition and structure of cell walls of biomass. Grasses such as wheat, maize, and rice, as well as dedicated perennial bioenergy crops, like switchgrass, make up ~55% of biomass that can be produced in the United States. Grass cell walls have a different composition and patterning compared with dicotyledonous plants, including the well-studied model plant, Arabidopsis. This project identified genetic determinants of cell wall composition in grasses using both naturally occurring genetic variation of switchgrass and gene network reconstruction and functional assays in rice. In addition, the project linked functional data in rice and other species to switchgrass improvement efforts through curation of the most abundant class of regulators in the switchgrass genome. Characterizing natural diversity of switchgrass for variation in cell wall composition and properties, also known as quality, provides an unbiased avenue for identifying biologically viable diversity in switchgrass cell walls. To characterizing natural diversity, this project generated cell wall composition and enzymatic deconstruction data for ~450 genotypes of the Switchgrass Southern Association Collection (SSAC), a diverse collection composed of 36 switchgrass accessions from the southern U.S. distribution of switchgrass. Comparing these data with other measures of cell wall quality for the same samples demonstrated the complementary nature of the diverse characterization platforms now being used for biomass characterization. Association of the composition data with ~3.2K single nucleotide variant markers identified six significant single nucleotide variant markers co-associated with digestibility and another compositional trait. These markers might be used to select switchgrass genotypes with improved composition in breeding programs for biofuel and forage production. Because the SSAC continues to be characterized by collaborators in the bioenergy community, the data generated will be used to identify additional markers in higher resolution genotyping data to approach identifying the genes and alleles that cause natural variation in switchgrass cell wall quality. For example, these markers can be surveyed in the 2100-member Oklahoma Southern and Northern Lowland switchgrass collections that this project also characterized. An orthogonal approach to biodiversity studies, using comparative functional genomics permits systematic querying of how much regulatory information is likely to be transferable from dicots to grasses and use of accumulated functional genomics resources for better-characterized grass species, such as rice, itself a biomass source in global agriculture and in certain regions. The project generated and tested a number of specific hypotheses regarding cell wall transcription factors and enzymes of grasses. To aid identification of cell wall regulators, the project assembled a novel, highdepth and -quality gene association network using a general linearized model scoring system to combine rice gene network data. Using known or putative orthologs of Arabidopsis cell wall biosynthesis genes and regulators, the project pulled from this network a cell wall sub-network that includes 96 transcription factors. Reverse genetics of a co-ortholog of the Arabidopsis MYB61 transcription factor in rice revealed that this regulatory node has evolved the ability to regulate grass-specific cell wall synthesis enzymes. A transcription factor with such activity has not been previously characterized to our knowledge, representing a major conclusion of this work. Changes in gene expression in a protoplast-based assay demonstrated positive or negative roles in cell wall regulation for eleven other transcription factors from the rice gene network. Eight of fifteen (53%) of these have not previously been examined for this function. Some of these may represent novel grass-diverged cell wall regulators, while others are likely to have this function across angiosperms. A parallel effort of this project to expand knowledge of enzymes that have evolved to function in grass cell wall synthesis, revealed that a grass-diverged enzyme in rice, OsAT 5 , ferulates monolignols that are naturally incorporated into grass cell walls. This finding opens potential natural selection avenues for improving biomass composition for downstream processing by weak base pretreatment. Thus, this project has significantly expanded knowledge of cell wall synthesis and regulation in rice, information that can be used in reverse genetics and synthetic biology approaches to re-engineer cell walls for improved production of biofuel and high-value products. To lay the foundation for translating these results directly for switchgrass improvement, the project employed a comparative phylogenetic analysis of the major group of cell wall transcription factors that have been found to function in cell wall regulation, the R 2 R 3 MYBs. This analysis concluded that known cell wall regulators are largely conserved across switchgrass, rice, maize, poplar, and Arabidopsis. This interpretation is also largely consistent with the gene network analysis described above, though both approaches provide evidence that some co-orthologs of Arabidopsis regulators have diminished or increased in importance based on gene expression patterns. Also, several clades containing dicot cell wall regulators have expanded, consistent with the evolution of new cell wall regulators. This latter result is supported by functional analysis of the R 2 R 3 MYB protein SWAM 1 in a collaboration between this project and the DOE-funded group of Dr. S. Hazen at the University of Massachusettes. The curation of the switchgrass genome through this project provides specific targets for future engineering of switchgrass cell wall regulation and may also facilitate identification of regulators that underlie the molecular markers that are genetically linked to differences in cell wall quality. With the goal of spurring further research and technological developments in lignocellulosic biofuel production, this work has been communicated to the bioenergy and cell wall communities though various presentations and publications. To date, three manuscripts have been published, two others are near to publication, three others are in an advanced state, and two to four more are likely to be written based on analyses still in progress. In addition, project participants have presented thirteen posters and talks at regional, national, and international meetings about aspects of this project. In sum, the work supported by this funding has made and communicated significant progress in identifying the genes that grasses use for cell wall synthesis and regulation, information that will be used by project participants and others to improve the efficiency of conversion of lignocellulosic biomass to biofuels.
Switchgrass cultivar EG1101
A switchgrass cultivar designated EG1101 is disclosed. Also disclosed are seeds of switchgrass cultivar EG1101, plants of switchgrass EG1101, plant parts of switchgrass cultivar EG1101 and methods for producing a switchgrass plant produced by crossing switchgrass cultivar EG1101 with itself or with another switchgrass variety. Methods are also described for producing a switchgrass plant containing in its genetic material one or more transgenes and to the transgenic switchgrass plants and plant parts produced by those methods. Switchgrass cultivars or breeding cultivars and plant parts derived from switchgrass variety EG1101, methods for producing other switchgrass cultivars, lines or plant parts derived from switchgrass cultivar EG1101 and the switchgrass plants, varieties, and their parts derived from use of those methods are described herein. Hybrid switchgrass seeds, plants and plant parts produced by crossing the cultivar EG1101 with another switchgrass cultivar are also described.
Switchgrass cultivar EG1102
A switchgrass cultivar designated EG1102 is disclosed. The invention relates to the seeds of switchgrass cultivar EG1102, to the plants of switchgrass EG1102, to plant parts of switchgrass cultivar EG1102 and to methods for producing a switchgrass plant produced by crossing switchgrass cultivar EG1102 with itself or with another switchgrass variety. The invention also relates to methods for producing a switchgrass plant containing in its genetic material one or more transgenes and to the transgenic switchgrass plants and plant parts produced by those methods. This invention also relates to switchgrass cultivars or breeding cultivars and plant parts derived from switchgrass variety EG1102, to methods for producing other switchgrass cultivars, lines or plant parts derived from switchgrass cultivar EG1102 and to the switchgrass plants, varieties, and their parts derived from use of those methods. The invention further relates to hybrid switchgrass seeds, plants and plant parts produced by crossing the cultivar EG1102 with another switchgrass cultivar.
Development of a Low Input and sustainable Switchgrass Feedstock Production System Utilizing Beneficial Bacterial Endophytes
Switchgrass represents a promising feedstock crop for US energy sustainability. However, its broad utilization for bioenergy requires improvements of biomass yields and stress tolerance. In this DOE funded project, we have been working on harnessing beneficial bacterial endophytes to enhance switchgrass performance and to develop a low input feedstock production system for marginal lands that do not compete with the production of food crops. We have demonstrated that one of most promising plant growth-promoting bacterial endophytes, Burkholderia phytofirmans strain PsJN, is able to colonize roots and significantly promote growth of switchgrass cv. Alamo under in vitro, growth chamber, greenhouse, as well as field conditions. Furthermore, PsJN bacterization improved growth and development of switchgrass seedlings, significantly stimulated plant root and shoot growth, and tiller number in the field, and enhanced biomass accumulation on both poor (p<0.001) and rich (p<0.05) soils, with more effective stimulation of plant growth in low fertility soil. Plant physiology measurements showed that PsJN inoculated Alamo had consistently lower transpiration, lower stomatal conductance, and higher water use efficiency in greenhouse conditions. These physiological changes may significantly contribute to the recorded growth enhancement. PsJN inoculation rapidly results in an increase in photosynthetic rates which contributes to the advanced growth and development. Some evidence suggests that this initial growth advantage decreases with time when resources are not limited such as in greenhouse studies. Additionally, better drought resistance and drought hardening were observed in PsJN inoculated switchgrass. Using the DOE-funded switchgrass EST microarray, in a collaboration with the Genomics Core Facility at the Noble Foundation, we have determined gene expression profile changes in both responsive switchgrass cv. Alamo and non-responsive cv. Cave-in-Rock (CR) following PsJN bacterization. With the MapMan software to analyze microarray data, the number of up- and down-regulated probes was calculated. The number of up-regulated probes in Alamo was 26, 14, 14, and 12% at 0.5, 2, 4 and 8 days after inoculation (DAI) with PsJN, respectively while the corresponding number in CR was 24, 22, 21, and 19%, respectively. In both cultivars, the largest number of up-regulated probes occurred at 0.5 DAI. Noticeable differences throughout the timeframe between Alamo and CR were that the number was dramatically decreased to half (12%) in Alamo but remained high in CR (approximately 20%). The number of down regulated genes demonstrated different trends in Alamo and CR. Alamo had an increasing trend from 9% at 0.5 DAI to 11, 17, and 28% at 2, 4, and 8 DAI, respectively. However, CR had 13% at 0.5 and 2 DAI, and declined to 10% at 4 and 8 DAI. With the aid of MapMan and PageMan, we mapped the response of the ID probes to the observed major gene regulatory network and major biosynthetic pathway changes associated with the beneficial bacterial endophyte infection, colonization, and early growth promotion process. We found significant differences in gene expression patterns between responsive and non-responsive cultivars in many pathways, including redox state regulation, signaling, proteolysis, transcription factors, as well as hormone (SA and JA in particular)-associated pathways. Form microarray data, a total of 50 key genes have been verified using qPCR. Ten of these genes were chosen for further functional study via either overexpression and/or RNAi knockout technologies. These genes were calmodulin-related calcium sensor protein (CAM), glutathione S-transferase (GST), histidine-containing phosphotransfer protein (H-221), 3 different zinc finger proteins (ZF-371, ZF131 and ZF242), EF hand transcription factor (EF-622), peroxidase, cellulose synthase catalytic submit A2 (CESA2), and Aux/IAA family. A total of 8 overexpression and 5 RNAi transgenic plants have been regenerated, and their gene expression levels determined using qPCR. Consequently high, medium and low expression lines were propagated in vitro for gene function study. When adequate numbers of individual transgenic lines were obtained, they were challenged with PsJN to see if PsJN promotes or inhibits growth of transgenic plants. Our results demonstrated that EF-622 overexpression, ZF-371, GST, H-221 and CAM RNAi transgenic lines lost responses to PsJN, i.e. PsJN had no growth promotive effects on these transgenic plants. Further study needs to be done to characterize this loss of responsiveness to PsJN. During this funding period, we have done more work related to this funded project and established collaborations with other institutions and obtained some interesting results, building a foundation for further research projects. For example, we isolated a naturally-occurring bacterium from surface-sterilized switchgrass seeds, identified as a unique Panteoa agglomerans species, and named strain PaKM. PaKM has been proved to be an efficient growth promoter of switchgrass over a broad spectrum of genotypes and has potential in applications with low input and sustainable production systems on marginal lands. In collaboration with Dr. Shuijin Hu (North Carolina State University), we conducted experiments on how endophyte-inoculated switchgrass affects soil N and P availability and the number of AMF in roots. Our preliminary results showed that PsJN increased AMF infection of switchgrass roots, and enhanced soil N availability and soil N mineralization on a low nutrient field. Further study of this phenomenon on different soils, over longer time periods, is needed to assess its potential impact on the productivity and longevity of switchgrass stands.