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Data from: Crop-associated virus infection in a native perennial grass: reduction in plant fitness and dynamic patterns of virus detection

To understand the eco-evolutionary significance of plant viruses in nature, we must (i) quantify the effects of infection on plant fitness and (ii) recognize that native plants are increasingly exposed to crop-associated viruses. Studies of perennials are particularly needed: most of our knowledge of plant-virus interactions is from annuals, yet long-lived species dominate landscapes. Here we used aster models for life-history analysis and longitudinal measures of plant virus status to evaluate multi-year consequences of crop virus infection in a native perennial. We used Barley yellow dwarf virus acquired from wheat to inoculate seedlings of Panicum virgatum L. (switchgrass), a North American prairie grass. We grew inoculated and mock-inoculated individuals of two ecotypes for 3 years in the field. We measured plant size, infection status and fitness components. Aster modelling provided integrated multi-year measures of fitness. Crop virus inoculation reduced multi-year native plant fitness by 30% over 2 years despite generally asymptomatic infection and evidence of resistance. This reduction was greater than predicted from individual fitness components or most size measures. Ecotypes differed in response, with the lowland ecotype experiencing higher apparent recovery from infection. Virus treatment in the upland ecotype delayed flowering phenology and reduced seed filling. Synthesis. Our use of field experimentation, surveys of plant infection status and aster modelling demonstrates a rigorous and broadly applicable approach for quantifying the effects of viruses and other microbes on multi-year plant fitness. We found that a crop virus had negative multi-year effects on native plant fitness even after infection was no longer detected. Viruses may have substantial effects on native vegetation with domestication of landscapes and agricultural expansion.

59 BASIC BIOLOGICAL SCIENCES↗

Empirical Evidence for the Potential Climate Benefits of Decarbonizing Light Vehicle Transport in the U.S. with Bioenergy from Purpose-Grown Biomass with and without BECCS

Climate mitigation scenarios limiting global temperature increases to 1.5 °C rely on decarbonizing vehicle transport with bioenergy production plus carbon capture and storage (BECCS), but climate impacts for producing different bioenergy feedstocks have not been directly compared experimentally or for ethanol vs electric light-duty vehicles. A field experiment at two Midwest U.S. sites on contrasting soils revealed that feedstock yields of seven potential bioenergy cropping systems varied substantially within sites but little between. Bioenergy produced per hectare reflected yields: miscanthus > poplar > switchgrass > native grasses ≈ maize stover (residue) > restored prairie ≈ early successional. Greenhouse gas emission intensities for ethanol vehicles ranged from 20 to -179 g CO 2 e MJ –1 : maize stover $\gg$ miscanthus ≈ switchgrass ≈ native grasses ≈ poplar > early successional ≥ restored prairie; direct climate benefits ranged from ~80% (stover) to 290% (restored prairie) reductions in CO 2 e compared to petroleum and were similar for electric vehicles. With carbon capture and storage (CCS), reductions in emission intensities ranged from 204% (stover) to 416% (restored prairie) for ethanol vehicles and from 329 to 558% for electric vehicles, declining 27 and 15%, respectively, once soil carbon equilibrates within several decades of establishment. Extrapolation based on expected U.S. transportation energy use suggests that, once CCS potential is maximized with CO 2 pipeline infrastructure, negative emissions from bioenergy with CCS for light-duty electric vehicles could capture >900 Tg CO 2 e year –1 in the U.S. In the future, as other renewable electricity sources become more important, electricity production from biomass would offset less fossil fuel electricity, and the advantage of electric over ethanol vehicles would decrease proportionately.

54 ENVIRONMENTAL SCIENCES↗

Climate cooling benefits of cellulosic bioenergy crops from elevated albedo

Abstract Changes in land surface albedo can alter ecosystem energy balance and potentially influence climate. We examined the albedo of six bioenergy cropping systems in southwest Michigan USA: monocultures of energy sorghum ( Sorghum bicolor ), switchgrass ( Panicum virgatum L.), and giant miscanthus ( Miscanthus × giganteus) , and polycultures of native grasses, early successional vegetation, and restored prairie. Direct field measurements of surface albedo ( α s ) from May 2018 through December 2020 at half‐hourly intervals in each system quantified the magnitudes and seasonal differences in albedo (∆ α ) and albedo‐induced radiative forcing (RF ∆ α ). We used a nearby forest as a historical native cover type to estimate reference albedo and RF ∆ α change upon original land use conversion, and a continuous no‐till maize ( Zea mays L .) system as a contemporary reference to estimate change upon conversion from annual row crops. Annually, α s differed significantly ( p < 0.05) among crops in the order: early successional (0.288 ± 0.012SE) >> miscanthus (0.271 ± 0.009) ≈ energy sorghum (0.270 ± 0.010) ≥ switchgrass (0.265 ± 0.009) ≈ restored prairie (0.264 ± 0.012) > native grasses (0.259 ± 0.010) > maize (0.247 ± 0.010). Reference forest had the lowest annual α s (0.134 ± 0.003). Albedo differences among crops during the growing season were also statistically significant, with growing season α s in perennial crops and energy sorghum on average ~20% higher (0.206 ± 0.003) than in no‐till maize (0.184 ± 0.002). Average non‐growing season (NGS) α s (0.370 ± 0.020) was much higher than growing season α s (0.203 ± 0.003) but these NGS differences were not significant. Overall, the original conversion of reference forest and maize landscapes to perennials provided a cooling effect on the local climate (RF αMAIZE : −3.83 ± 1.00 W m −2 ; RF αFOREST : −16.75 ± 3.01 W m −2 ). Significant differences among cropping systems suggest an additional management intervention for maximizing the positive climate benefit of bioenergy crops, with cellulosic crops on average ~9.1% more reflective than no‐till maize, which itself was about twice as reflective as the reference forest.

Lei, Cheyenne↗

Using Prescribed Fire to Manage the Fernald On-Site Disposal Facility Vegetated Cap - 17401

The Fernald Preserve is a former uranium processing plant that underwent extensive remediation pursuant to the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA). It is situated on a 425 ha (1,050 acre) tract of land, approximately 29 km (18 miles) northwest of Cincinnati, Ohio. Remedial activities and subsequent ecological restoration have converted the site from an industrial production facility to an undeveloped park, encompassing a series of wetlands, prairies, and forested communities. The remediation of buildings and soil contamination was completed in 2006, but aquifer remediation is ongoing. Since 2007, the site has been managed by the US Department of Energy (DOE) Office of Legacy Management. Portions of the site are open to the public, with approximately 11 km (7 miles) of walking trails that are open year-round and a visitors center. Site remediation included the construction of an On-Site Disposal Facility (OSDF) for the long-term disposal of contaminated soil and building materials. The OSDF consists of eight disposal cells that encompass approximately 30 hectares (75 acres) on the eastern portion of the Fernald site. Cells are lined and capped, and each has a separate leak detection and leachate collection system. The integrated cap is vegetated with native grasses and forbs. Management of the Fernald Preserve includes the monitoring and maintenance of ecologically restored communities and the vegetated OSDF cap, as required by the site's institutional controls plan. Prescribed burning is recognized as the preferred method for prairie management. Prescribed fire has been used at the Fernald site since 2009 for established prairies in restored areas, but not on the OSDF cap. Stakeholders had reservations regarding the safety of using prescribed fire on the OSDF cap. Because of that, vegetation on the cap was managed as a grassland community. It was mowed, raked, and baled to remove thatch, and promote the continued establishment of native grasses and forbs. Since agricultural use is restricted at the site, the resulting hay bales were kept on property and reused as organic matter and mulch. Over time, more hay bales were being generated in a season than were reused. Therefore in 2015, DOE decided to revisit stakeholder acceptance of prescribed fire on the OSDF cap. Ongoing relationships with local stakeholders allowed for extensive discussions regarding the current management and proposed use of prescribed fire on the OSDF cap. The public was provided information about the planning and implementation of prescribed burns at the site and invited to observe the execution of a successful site burn. This approach led to stakeholder acceptance of DOE's proposal, and vegetation on the caps of cells 4, 5, and 6 was burned in late March 2016. The burn was successful and resulted in several benefits from a land stewardship and cost perspective. This paper details the stakeholder engagement approach and the resulting benefits from the use of prescribed fire on the OSDF cap. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Flight Considerations and Hyperspectral Image Classifications for Dryland Vegetation Management from a Fixed-wing UAS

Unmanned Aerial Systems (UAS)-based hyperspectral remote sensing capabilities developed by the Idaho National Lab and Boise Center Aerospace Lab were tested via demonstration flights that explored the influence of altitude on geometric error, image mosaicking, and dryland vegetation classification. The motivation for this study was to better understand the challenges associated with UAS-based hyperspectral data for distinguishing native grasses such as Sandberg bluegrass ( Poa secunda ) from invasives such as burr buttercup ( Ranunculus testiculatus) in a shrubland environment. The test flights successfully acquired usable flightline data capable of supporting classifiable composite images. Unsupervised classification results support vegetation management objectives that rely on mapping shrub cover and distribution patterns. However, supervised classifications performed poorly despite spectral separability in the image-derived endmember pixels. In many cases, the supervised classifications accentuated noise or features in the mosaic that were artifacts of color balancing and feathering in areas of flightline overlap. Future UAS flight missions that optimize flight planning; minimize illumination differences between flightlines; and leverage ground reference data and time series analysis should be able to effectively distinguish native grasses such as Sandberg bluegrass from burr buttercup.

58 GEOSCIENCES↗

Data from: Early accumulation of active fraction soil carbon in newly established cellulosic biofuel systems

We examined relative changes in soil C pools shortly after the establishment of six perennial and two annual bioenergy cropping systems that differed in diversity (monoculture vs. polyculture). Perennial systems included two monocultures (switchgrass, Panicum virgatum; and miscanthus, Miscanthus × giganteus) and four polycultures including hybrid poplar (Populus sp.) + herbaceous understory; mixed native grasses, successional vegetation, and restored prairie. Two annual systems included no-till continuous corn (Zea mays) and rotational corn (corn-soybean (Glycine max)-canola (Brassica napus)). Each crop was planted in a full factorial design at both a moderate fertility Alfisol and a high fertility Mollisol site. Relative differences in active, slow, and passive C pools in surface soils, where C changes are most likely to be detected early, were evaluated with 322-day laboratory incubations followed by acid hydrolysis to infer different pools from exponential decay curves. Five years post-establishment, active C pools under perennial polycultures at the Alfisol site were up to twice those under annual and perennial monocultures, and followed the order hybrid poplars (696 ± 216 μg C g− 1 soil, n = 5 replicate blocks) ≈ native grasses (656 ± 155) ≈ restored prairie (638 ± 44) > early successional (500 ± 54) ≫ continuous corn (237 ± 68) ≈ rotational corn (180 ± n.a.). Active C pools in perennial monocultures were similar to those in continuous corn: switchgrass (274 ± 29) ≈ miscanthus (299 ± 9). In contrast, differences in active C pools among crops at the more fertile Mollisol site were not detectable except for greater pools in the restored prairie and rotational corn systems. At both sites, slow and passive C pools differed little among systems except that slow pools were greater in the poplar system. That diversity rather than perenniality itself led to greater active C pools suggests that polycultures might be used to accelerate soil C accumulation in bioenergy and other perennial cropping systems.

09 BIOMASS FUELS↗

Population structure of Miscanthus sacchariflorus reveals two major polyploidization events, tetraploid-mediated unidirectional introgression from diploid M. sinensis, and diversity centred around the Yellow Sea

Abstract Background and Aims Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sacchariflorus has a broad geographic range, is used to produce paper in China and is one of the parents (along with Miscanthus sinensis) of the important biomass species Miscanthus × giganteus. The largest study of M. sacchariflorus population genetics to date is reported here. Methods Collections included 764 individuals across East Asia. Samples were genotyped with 34 605 single nucleotide polymorphisms (SNPs) derived from restriction site-associated DNA sequencing (RAD-seq) and ten plastid microsatellites, and were subjected to ploidy analysis by flow cytometry. Key Results Six major genetic groups within M. sacchariflorus were identified using SNP data: three diploid groups, comprising Yangtze (M. sacchariflorus ssp. lutarioriparius), N China and Korea/NE China/Russia; and three tetraploid groups, comprising N China/Korea/Russia, S Japan and N Japan. Miscanthus sacchariflorus ssp. lutarioriparius was derived from the N China group, with a substantial bottleneck. Japanese and mainland tetraploids originated from independent polyploidization events. Hybrids between diploid M. sacchariflorus and M. sinensis were identified in Korea, but without introgression into either parent species. In contrast, tetraploid M. sacchariflorus in southern Japan and Korea exhibited substantial hybridization and introgression with local diploid M. sinensis. Conclusions Genetic data indicated that the land now under the Yellow Sea was a centre of diversity for M. sacchariflorus during the last glacial maximum, followed by a series of migrations as the climate became warmer and wetter. Overall, M. sacchariflorus has greater genetic diversity than M. sinensis, suggesting that breeding and selection within M. sacchariflorus will be important for the development of improved M. × giganteus. Ornamental M. sacchariflorus genotypes in Europe and North America represent a very narrow portion of the species’ genetic diversity, and thus do not well represent the species as a whole.

Clark, Lindsay V.↗

The La Mesa Fire: Studies and Observations from 1975 through 2012

In 1996, Swetnam and Baisan stated the following: “The 1977 La Mesa burn was a wake-up call to perhaps the most pressing forest health problem in Southwestern forests—historically anomalous, catastrophic wildfire in the ponderosa pine—created by many decades of fire exclusion.” However, the wake-up call was only momentary and was forgotten until the Dome Fire (1996), Oso Complex Fire (1998), and Cerro Grande Fire (2000) burned lands within the community of Los Alamos, Los Alamos National Laboratory (LANL), Bandelier National Monument (BNM), and Santa Fe National Forest (SFNF). After the Dome Fire, several initiatives were under way to understand the wildland-urban interface and the conditions that have brought the forests of the Pajarito Plateau dangerously close to multiple, large, and intense wildfires. After the Dome Fire, the Interagency Wildfire Management Team was formed, bringing together adjoining agencies to deal with the ever-increasing threat of conflagration that could result in loss of property and human life. This study is about the history before and after the La Mesa Fire—at the time, the first large wildfire on the Pajarito Plateau in over 80 years. It represents 37 years of research and observations that began when management of fire on the Pajarito Plateau was first being explored by BNM. It documents changes in vegetation composition since June 1977 and shows the progression of change in understanding fire’s essential place in the ecosystems of the Pajarito Plateau. The study began in 1975 and 1976, when Loren Potter and I examined previously burned areas to determine a fire frequency for the east Jemez Mountains. We had examined areas of 10 acres or more that had burned at various times, e.g., 1939, 1945, 1950, 1955, 1960, and 1976. A year and a half into the study, the La Mesa Fire ignited on Mesa del Rito, ultimately burning about 15,000 acres of BNM, SFNF, and LANL, including our established plots. After the fire, we set up 20- by 50-meter permanent plots along four ornithological transects established by ornithologist Roland Wauer. Wauer’s transects were mostly in ponderosa pine, with one in mixed conifer and one in the piñon-juniper woodland. Plots within the ponderosa pine were visited in 1977, 1978, 1985, 1992, 1998, 1999, 2010, 2011 and 2012. We mapped the standing trees, photographed each plot, and collected data on the understory components. Transects in the mixed conifer were visited in 1977, 1992, 1999, 2010 and 2012. Plots in the piñon-juniper were visited in 1977, 1999, and 2010 for taking photographs and data years after the fire. Additionally, plots were established in LANL Technical Area 49 as a control to post-fire seeding. We visited these plots in 1977, 1992, and 1998. Ironically, this study ends with another large fire burning over the plots we have followed all these years. In June 2011, the Las Conchas Fire became the largest fire in New Mexico’s history to that time and burned most of the area of the La Mesa Fire and the plots studied over the years. In this report, we summarize information about the vegetation changes within the boundaries of the La Mesa Fire. Thirty-seven years of observations and studies have provided long-term data related to regeneration of trees, water relations of grasses and pines, the influence of seeding with non-native grasses, pyrodendrochronology, and plant succession as related to fire.

54 ENVIRONMENTAL SCIENCES↗

AmeriFlux US-MCP Moro Cojo Slough, PG&E Treatment Wetland

This is the AmeriFlux version of the carbon flux data for the site US-MCP Moro Cojo Slough, PG&E Treatment Wetland. Site Description - Freshwater coastal wetland, restored in 2016, native grasses and sedges among freshwater ponds, surrounded 400m in each direction by agricultural plots, native plants remove excess agrcultural nitrogen from the soil

Paytan, Adina [University of California, Santa Cru↗

Interactions between Bacillus anthracis and Plants May Promote Anthrax Transmission

Environmental reservoirs are essential in the maintenance and transmission of anthrax but are poorly characterized. The anthrax agent, Bacillus anthracis was long considered an obligate pathogen that is dormant and passively transmitted in the environment. However, a growing number of laboratory studies indicate that, like some of its close relatives, B. anthracis has some activity outside of its vertebrate hosts. Here we show in the field that B. anthracis has significant interactions with a grass that could promote anthrax spore transmission to grazing hosts. Using a local, virulent strain of B. anthracis, we performed a field experiment in an enclosure within a grassland savanna. We found that B. anthracis increased the rate of establishment of a native grass (Enneapogon desvauxii) by 50% and that grass seeds exposed to blood reached heights that were 45% taller than controls. Further we detected significant effects of E. desvauxii, B. anthracis, and their interaction on soil bacterial taxa richness and community composition. We did not find any evidence for multiplication or increased longevity of B. anthracis in bulk soil associated with grass compared to controls. Instead interactions between B. anthracis and plants may result in increased host grazing and subsequently increased transmission to hosts.

54 ENVIRONMENTAL SCIENCES↗

Nitrate Leaching from Continuous Corn, Perennial Grasses, and Poplar in the US Midwest

Leaching from annual corn ( Zea mays L.) crops is a primary source of nitrate (NO 3 - ) pollution of ground and surface waters. Here, we compare NO 3 - losses from no-till corn with losses from various alternative perennial cropping systems (switchgrass [ Panicum virgatum L.], miscanthus [ Miscanthus ×giganteus J.M. Greef & Deuter ex Hodkinson & Renvoiz], a native grass mixture, and restored prairie), as well as hybrid poplar ( Populus nigra L. × P. maximowiczii A. Henry ‘NM6’), all grown on a well-drained soil in Michigan. Soil water was sampled from below the root zone using suction cup samplers during nonfrozen periods (March–November) between 2009 and 2016. Leaching was estimated from NO 3 - concentrations in soil water and modeled drainage (percolation) rates. Drainage rates were not significantly different among crops, constituting ~30% of total annual precipitation. Aboveground net primary production (Mg ha -1 yr -1 ) averaged across the 7 yr was highest in poplar (30.8 ± 1.9 [SE]) followed by miscanthus (23.9 ± 2.4) and corn (20.4 ± 0.9). Volume-weighted mean NO 3 - concentrations (mg N L -1 ) and NO 3 - leaching (kg ha -1 yr -1 ) averaged across the 7 yr were 9.2 and 34.1, 2.3 and 5.9, and 3.0 and 7.2, respectively, for corn, perennial grasses and poplar. Approximately 10 to 32% of applied N was lost as NO 3 - from these crops, with the highest percent losses from poplar (32%) followed by corn (20%). Perennial cropping systems leached considerably more NO 3 - in first few years after planting, but over 7 yr they lost much less NO 3 - than corn. Perennial crops may therefore help ameliorate NO 3 - pollution in agricultural landscapes even if they receive modest N fertilization.

54 ENVIRONMENTAL SCIENCES↗

Nitrogen Fertilization Effects on Productivity and Nitrogen Loss in Three Grass-Based Perennial Bioenergy Cropping Systems

Nitrogen (N) fertilization can greatly improve plant productivity but needs to be carefully managed to avoid harmful environmental impacts. Nutrient management guidelines aimed at reducing harmful forms of N loss such as nitrous oxide (N 2 O) emissions and nitrate (NO 3 - ) leaching have been tailored for many cropping systems. The developing bioenergy industry is likely to make use of novel cropping systems, such as polycultures of perennial species, for which we have limited nutrient management experience. We studied how a switchgrass (Panicum virgatum) monoculture, a 5-species native grass mixture and an 18- species restored prairie responded to annual fertilizer applications of 56 kg N ha -1 in a fieldscale agronomic trial in south-central Wisconsin over a 2-year period.We observed greater fertilizer-induced N 2 O emissions and sub-rooting zone NO 3 - concentrations in the switchgrass monoculture than in either polyculture. Fertilization increased aboveground net primary productivity in the polycultures, but not in the switchgrass monoculture. Switchgrass was generally more productive, while the two polycultures did not differ from each other in productivity or N loss. In conclusion, our results highlight differences between polycultures and a switchgrass monoculture in responding to N fertilization.

54 ENVIRONMENTAL SCIENCES↗

Biomass Production a Stronger Driver of Cellulosic Ethanol Yield than Biomass Quality

Core Ideas Fermentable sugars were greatest in corn stover > perennial grasses > polycultures. Corn stover had the highest ethanol content. Miscanthus had the highest ethanol yield potential on a per hectare basis. Ethanol yield potential per hectare of switchgrass ≥ corn stover. Biomass yield was the strongest driver of per hectare ethanol yield. Many crops have been proposed as feedstocks for the emerging cellulosic ethanol industry, but information is lacking about the relative importance of feedstock production and quality. We compared yield and sugar content for seven bioenergy cropping systems in south‐central Wisconsin (ARL) and southwestern Michigan (KBS) during three growing seasons (2012 through 2014). The cropping systems were (i) continuous corn stover ( Zea mays L.), (ii) switchgrass ( Panicum virgatum L.), (iii) giant miscanthus ( Miscanthus × giganteus Greef & Deuter ex Hodkinson & Renvoize), (iv) hybrid poplar ( Populus nigra × P. maximowiczii A. Henry ‘NM6’), (v) native grass mix, (vi) early successional community, and (vii) restored prairie. A high‐throughput pretreatment and fermentation assay showed corn stover with the highest sugar content (213 g glucose kg −1 [Glc] and 115 g xylose kg −1 [Xyl]) followed by the two monoculture perennial grass treatments (154 [Glc] and 88 [Xyl]) and then the herbaceous polycultures (135 [Glc] and 77 [Xyl]). Biomass production and sugar content were combined to calculate ethanol yields. Miscanthus had the highest per hectare ethanol yields (1957 l ha −1 yr −1 ARL, 2485 l ha −1 yr −1 KBS) followed by switchgrass (1091 l ha −1 yr −1 ARL, 1017 l ha −1 yr −1 KBS) and corn stover (1121 l ha −1 yr −1 ARL, 878 l ha −1 yr −1 KBS). Perennial grass cropping systems (i.e., switchgrass and miscanthus) had higher per hectare ethanol yields at both sites relative to diverse systems that included dicots. Despite feedstock differences in fermentable sugars, biomass production was the strongest driver of per hectare ethanol yield.

09 BIOMASS FUELS↗

Expanding the breeder’s toolbox for perennial grasses

With the increase in world energy demand and the environmental impacts associated with continued fossil fuel use, there is a great need for the development of alternative and sustainable fuel sources. Switchgrass ( Panicum virgatum L. ), a perennial grass native to North America, is poised to become one of the first dedicated biomass crops deployed on a large scale in the United States in large part due to its inherent high yield, adaptability and perennial nature. Current ‘cultivars’ of switchgrass are essentially undomesticated wild selections from natural populations. Thus, there is considerable potential for improvement of this emerging energy crop. Traditional breeding approaches in switchgrass have been hampered by its self-incompatibility and sensitivity to inbreeding, therefore there is a significant need to develop new tools for improved switchgrass breeding. The proposed research uses a systems biology approach to accelerate switchgrass breeding, to develop its biotechnological improvement through the generation of stress-tolerant varieties, and to establish ways to minimize the potential impact of transgenes on ecosystems. In this proposal, we will develop new breeding tools for perennial grasses and apply these tools toward improving switchgrass. Specifically, we will employ two complementary approaches to overcome the difficulty of breeding a large, polyploid, self-incompatible plant. The first approach will accelerate conventional breeding by developing a method to rapidly create doubled haploid (homozygous) lines for breeding. This approach is based on altering CENH3, a centromere-specific histone, leading to the production of haploid offspring when the centromere mutant is crossed to wild type plants. Subsequent genome doubling results in fully homozygous individuals and overcomes one of the major limitations in breeding self-incompatibile crops. These homozygous lines would allow switchgrass breeders to apply a variety of breeding approaches that have been previously impossible. The second approach is to develop tools and reagents to accelerate the biotechnological improvement of switchgrass including: modules for improved stress tolerance, a test bed to evaluate transgenes in a perennial model grass and a robust gene containment system. In order to maintain a favorable energy balance and avoid competition with food crops, dedicated biomass crops must be grown with minimal inputs (fertilizer and water) on marginal lands that are unsuited to the traditional high input agriculture necessary to produce food. Thus, abiotic stress tolerance and efficient water and nutrient use are of paramount importance. Considerable basic research has been directed at these areas and this knowledge has been used to develop transgenic approaches to greatly improve water and nutrient use efficiency. By applying these approaches to switchgrass we will rapidly increase drought tolerance and nutrient use efficiency. This approach can rapidly introduce these traits in different cultivars allowing breeders to add these traits to their elite germplasm. Since biotechnologically improved switchgrass must ultimately be grown in open fields, they must be designed to minimize the potential impact of the transgenes on non-target ecosystems. The best way to do this is to minimize gene flow from the transgenic plants. Thus, we will develop technology to introduce sterility into the transgenic plants. We will then combine this technology with the stress resistance modules to create switchgrass lines with enhanced resistance to abiotic stresses.

09 BIOMASS FUELS↗

Expanding the Breeder’s Toolbox for Perennial Grasses

With the increase in world energy demand and the environmental impacts associated with continued fossil fuel use, there is a great need for the development of alternative and sustainable fuel sources. Switchgrass (Panicum virgatum L.), a perennial grass native to North America, is poised to become one of the first dedicated biomass crops deployed on a large scale in the United States in large part due to its inherent high yield, adaptability and perennial nature. Current ‘cultivars’ of switchgrass are essentially undomesticated wild selections from natural populations. Thus, there is considerable potential for improvement of this emerging energy crop. Traditional breeding approaches in switchgrass have been hampered by its self-incompatibility and sensitivity to inbreeding, therefore there is a significant need to develop new tools for improved switchgrass breeding. The proposed research uses a systems biology approach to accelerate switchgrass breeding, to develop its biotechnological improvement through the generation of stress-tolerant varieties, and to establish ways to minimize the potential impact of transgenes on ecosystems. In this proposal, we will develop new breeding tools for perennial grasses and apply these tools toward improving switchgrass. Specifically, we will employ two complementary approaches to overcome the difficulty of breeding a large, polyploid, self-incompatible plant. The first approach will accelerate conventional breeding by developing a method to rapidly create doubled haploid (homozygous) lines for breeding. This approach is based on altering CENH3, a centromere-specific histone, leading to the production of haploid offspring when the centromere mutant is crossed to wild type plants. Subsequent genome doubling results in fully homozygous individuals and overcomes one of the major limitations in breeding self-incompatibile crops. These homozygous lines would allow switchgrass breeders to apply a variety of breeding approaches that have been previously impossible. The second approach is to develop tools and reagents to accelerate the biotechnological improvement of switchgrass including: modules for improved stress tolerance, a test bed to evaluate transgenes in a perennial model grass and a robust gene containment system. In order to maintain a favorable energy balance and avoid competition with food crops, dedicated biomass crops must be grown with minimal inputs (fertilizer and water) on marginal lands that are unsuited to the traditional high input agriculture necessary to produce food. Thus, abiotic stress tolerance and efficient water and nutrient use are of paramount importance. Considerable basic research has been directed at these areas and this knowledge has been used to develop transgenic approaches to greatly improve water and nutrient use efficiency. By applying these approaches to switchgrass we will rapidly increase drought tolerance and nutrient use efficiency. This approach can rapidly introduce these traits in different cultivars allowing breeders to add these traits to their elite germplasm. Since biotechnologically improved switchgrass must ultimately be grown in open fields, they must be designed to minimize the potential impact of the transgenes on non-target ecosystems. The best way to do this is to minimize gene flow from the transgenic plants. Thus, we will develop technology to introduce sterility into the transgenic plants. We will then combine this technology with the stress resistance modules to create switchgrass lines with enhanced resistance to abiotic stresses.

09 BIOMASS FUELS↗

Inter-Annual Precipitation Variability Decreases Switchgrass Productivity from Arid to Mesic Environments

Cellulosic biofuels are an important source of renewable biomass within the alternative energy portfolio. Switchgrass (Panicum virgatum L.), a perennial C 4 grass native to North America, is widely studied as a biofuel feedstock for its consistently high yields and minimal input requirements. The influences of precipitation amount and temporal variability on the fertilizer response of switchgrass productivity are not fully understood. Moreover, global climate models predict changes in rainfall patterns towards lower and increasingly variable soil water availability in several productive areas worldwide, which may impact net primary production of biofuel crops. We conducted a meta-analysis of aboveground net primary production of switchgrass from 48 publications encompassing 82 different locations, 11 soil types, 52 switchgrass cultivars, fertilizer inputs between 0 to 896 kg N ha –1 year –1 , and 1 to 6 years of annual productivity measures repeated on the same stand. Productivity of the lowland ecotype doubled with N rates > 131 kg N ha –1 year –1 , but upland ecotype productivity increased only by 50%. Results showed an optimum N rate of 30 to 60 kg N ha –1 year –1 for both ecotypes, after which biomass gain per unit of N added decreased. Growing season precipitation (GSPPT) and inter-annual precipitation variability (inter-PPTvar) affected both ecotypes similarly. Long-term mean annual precipitation (MAP) differentially affected lowland and upland productivity, depending on the N level. Productivity responses to MAP and GSPPT were similar for both upland and lowland ecotypes at none or low N rates. When N increased beyond 60 kg N ha –1 year –1 , lowland cultivars had a greater growth response to MAP than uplands. Productivity increased with increasing GSPPT and MAP and had a positive linear response to MAP ranging from 600 to 1200 mm year –1 . One third of the variability in switchgrass production was accounted for by inter-PPTvar. After accounting for MAP, sites with higher inter-PPTvar had lower switchgrass productivity than sites with lower inter-PPTvar. Increased inter-annual variation in precipitation reduced production of both ecotypes. As a result, predicted changes in the amount and timing of precipitation thus likely will exert greater influence on production of upland than lowland ecotypes of switchgrass.

54 ENVIRONMENTAL SCIENCES↗