Restoration Handbook for Sagebrush Steppe Ecosystems with Emphasis on Greater Sage-Grouse Habitat—Part 3. Site Level Restoration Decisions

Sagebrush steppe ecosystems in the United States currently (2016) occur on only about one-half of their historical land area because of changes in land use, urban growth, and degradation of land, including invasions of non-native plants. The existence of many animal species depends on the existence of sagebrush steppe habitat. The greater sage-grouse (Centrocercus urophasianus) depends on large landscapes of intact habitat of sagebrush and perennial grasses for their existence. In addition, other sagebrush-obligate animals have similar requirements and restoration of landscapes for greater sage-grouse also will benefit these animals. Once sagebrush lands are degraded, they may require restoration actions to make those lands viable habitat for supporting sagebrush-obligate animals, livestock, and wild horses, and to provide ecosystem services for humans now and for future generations. When a decision is made on where restoration treatments should be applied, there are a number of site-specific decisions managers face before selecting the appropriate type of restoration. This site-level decision tool for restoration of sagebrush steppe ecosystems is organized in nine steps. ●Step 1 describes the process of defining site-level restoration objectives. ●Step 2 describes the ecological site characteristics of the restoration site. This covers soil chemistry and texture, soil moisture and temperature regimes, and the vegetation communities the site is capable of supporting. ●Step 3 compares the current vegetation to the plant communities associated with the site State and Transition models. ●Step 4 takes the manager through the process of current land uses and past disturbances that may influence restoration success. ●Step 5 is a brief discussion of how weather before and after treatments may impact restoration success. ●Step 6 addresses restoration treatment types and their potential positive and negative impacts on the ecosystem and on habitats, especially for greater sage-grouse. We discuss when passive restoration options may be sufficient and when active restoration may be necessary to achieve restoration objectives. ●Step 7 addresses decisions regarding post-restoration livestock grazing management. ●Step 8 addresses monitoring of the restoration; we discuss important aspects associated with implementation monitoring as well as effectiveness monitoring. ●Step 9 takes the information learned from monitoring to determine how restoration actions in the future might be adapted to improve restoration success

Restoration Handbook for Sagebrush Steppe Ecosystems with Emphasis on Greater Sage-Grouse Habitat—Part 1. Concepts for Understanding and Applying Restoration

Sagebrush steppe ecosystems in the United States currently occur on only about one-half of their historical land area because of changes in land use, urban growth, and degradation of land, including invasions of non-native plants. The existence of many animal species depends on the existence of sagebrush steppe habitat. The greater sage-grouse (Centrocercus urophasianus) is a landscape-dependent bird that requires intact habitat and combinations of sagebrush and perennial grasses to exist. In addition, other sagebrush-obligate animals also have similar requirements and restoration of landscapes for greater sage-grouse also will benefit these animals. Once sagebrush lands are degraded, they may require restoration actions to make those lands viable habitat for supporting sagebrush-obligate animals. This restoration handbook is the first in a three-part series on restoration of sagebrush ecosystems. In Part 1, we discuss concepts surrounding landscape and restoration ecology of sagebrush ecosystems and greater sage-grouse that habitat managers and restoration practitioners need to know to make informed decisions regarding where and how to restore specific areas. We will describe the plant dynamics of sagebrush steppe ecosystems and their responses to major disturbances, fire, and defoliation. We will introduce the concepts of ecosystem resilience to disturbances and resistance to invasions of annual grasses within sagebrush steppe. An introduction to soils and ecological site information will provide insights into the specific plants that can be restored in a location. Soil temperature and moisture regimes are described as a tool for determining resilience and resistance and the potential for various restoration actions. Greater sage-grouse are considered landscape birds that require large areas of intact sagebrush steppe; therefore, we describe concepts of landscape ecology that aid our decisions regarding habitat restoration. We provide a brief overview of restoration techniques for sage-grouse habitat restoration. We conclude with a description of the critical nature of monitoring for adaptive management of sagebrush steppe restoration at landscape- and project-specific levels

Restoration Handbook for Sagebrush Steppe Ecosystems with Emphasis on Greater Sage-Grouse Habitat—Part 2. Landscape Level Restoration Decisions

Sagebrush steppe ecosystems in the United States currently (2015) occur on only about one-half of their historical land area because of changes in land use, urban growth, and degradation of land, including invasions of non-native plants. The existence of many animal species depends on the existence of sagebrush steppe habitat. The greater sage-grouse (Centrocercus urophasianus) is a landscape-dependent bird that requires intact habitat and combinations of sagebrush and perennial grasses to exist. In addition, other sagebrush-obligate animals also have similar requirements and restoration of landscapes for greater sage-grouse also will benefit these animals. Once sagebrush lands are degraded, they may require restoration actions to make those lands viable habitat for supporting sagebrush-obligate animals. Land managers do not have resources to restore all locations because of the extent of the restoration need and because some land uses are not likely to change, therefore, restoration decisions made at the landscape to regional scale may improve the effectiveness of restoration to achieve landscape and local restoration objectives. We present a landscape restoration decision tool intended to assist decision makers in determining landscape objectives, to identify and prioritize landscape areas where sites for priority restoration projects might be located, and to aid in ultimately selecting restoration sites guided by criteria used to define the landscape objectives. The landscape restoration decision tool is structured in five sections that should be addressed sequentially. Each section has a primary question or statement followed by related questions and statements to assist the user in addressing the primary question or statement. This handbook will guide decision makers through the important process steps of identifying appropriate questions, gathering appropriate data, developing landscape objectives, and prioritizing landscape patches where potential sites for restoration projects may be located. Once potential sites are selected, land managers can move to the site-specific decision tool to guide restoration decisions at the site level

A Synopsis of Short-Term Response to Alternative Restoration Treatments in Sagebrush-Steppe: The SageSTEP Project

AbstractThe Sagebrush Steppe Treatment Evaluation Project (SageSTEP) is an integrated long-term study that evaluates ecological effects of alternative treatments designed to reduce woody fuels and to stimulate the herbaceous understory of sagebrush steppe communities of the Intermountain West. This synopsis summarizes results through 3 yr posttreatment. Woody vegetation reduction by prescribed fire, mechanical treatments, or herbicides initiated a cascade of effects, beginning with increased availability of nitrogen and soil water, followed by increased growth of herbaceous vegetation. Response of butterflies and magnitudes of runoff and erosion closely followed herbaceous vegetation recovery. Effects on shrubs, biological soil crust, tree cover, surface woody fuel loads, and sagebrush-obligate bird communities will take longer to be fully expressed. In the short term, cool wet sites were more resilient than warm dry sites, and resistance was mostly dependent on pretreatment herbaceous cover. At least 10 yr of posttreatment time will likely be necessary to determine outcomes for most sites. Mechanical treatments did not serve as surrogates for prescribed fire in how each influenced the fuel bed, the soil, erosion, and sage-obligate bird communities. Woody vegetation reduction by any means resulted in increased availability of soil water, higher herbaceous cover, and greater butterfly numbers. We identified several trade-offs (desirable outcomes for some variables, undesirable for others), involving most components of the study system. Trade-offs are inevitable when managing complex natural systems, and they underline the importance of asking questions about the whole system when developing management objectives. Substantial spatial and temporal heterogeneity in sagebrush steppe ecosystems emphasizes the point that there will rarely be a “recipe” for choosing management actions on any specific area. Use of a consistent evaluation process linked to monitoring may be the best chance managers have for arresting woodland expansion and cheatgrass invasion that may accelerate in a future warming climate

A Synopsis of Short-Term Response to Alternative Restoration Treatments in Sagebrush-Steppe: The SageSTEP Project

The Sagebrush Steppe Treatment Evaluation Project (SageSTEP) is an integrated long-term study that evaluates ecological effects of alternative treatments designed to reduce woody fuels and to stimulate the herbaceous understory of sagebrush steppe communities of the Intermountain West. This synopsis summarizes results through 3 yr posttreatment. Woody vegetation reduction by prescribed fire, mechanical treatments, or herbicides initiated a cascade of effects, beginning with increased availability of nitrogen and soil water, followed by increased growth of herbaceous vegetation. Response of butterflies and magnitudes of runoff and erosion closely followed herbaceous vegetation recovery. Effects on shrubs, biological soil crust, tree cover, surface woody fuel loads, and sagebrush-obligate bird communities will take longer to be fully expressed. In the short term, cool wet sites were more resilient than warm dry sites, and resistance was mostly dependent on pretreatment herbaceous cover. At least 10 yr of posttreatment time will likely be necessary to determine outcomes for most sites. Mechanical treatments did not serve as surrogates for prescribed fire in how each influenced the fuel bed, the soil, erosion, and sage-obligate bird communities. Woody vegetation reduction by any means resulted in increased availability of soil water, higher herbaceous cover, and greater butterfly numbers. We identified several trade-offs (desirable outcomes for some variables, undesirable for others), involving most components of the study system. Trade-offs are inevitable when managing complex natural systems, and they underline the importance of asking questions about the whole system when developing management objectives. Substantial spatial and temporal heterogeneity in sagebrush steppe ecosystems emphasizes the point that there will rarely be a “recipe” for choosing management actions on any specific area. Use of a consistent evaluation process linked to monitoring may be the best chance managers have for arresting woodland expansion and cheatgrass invasion that may accelerate in a future warming climate

A REGIONAL EXPERIMENT TO EVALUATE EFFECTS OF FIRE AND FIRE SURROGATE TREATMENTS IN THE SAGEBRUSH BIOME

SageSTEP is a comprehensive regional experiment that provides critical information to managers faced with a sagebrush steppe ecosystem that is increasingly at risk from wildfire, invasive plants, and climate change. The experiment provides managers with information that can be used to restore ecological communities across the 100+ million acres of the sagebrush biome. It is designed to match the temporal and spatial scales at which managers operate, is intended to reduce management risk and uncertainty of catastrophic wildfire to the greatest degree possible, and provides managers with information that allows them to better understand tradeoffs inherent in the choice of management alternatives. The project has several features that make it ideal for testing hypotheses from state‐andtransition theory, and for discovering information that can be directly applied in a management context ‐‐ it is long‐term, experimental, multisite, multivariate, and treatments are applied across condition gradients, allowing for potential identification of biotic thresholds. The project is designed to distinguish communities that have conditions that will allow them to recover on their own following fuel or restoration treatments, versus communities that have crossed biotic thresholds, and will therefore require more expensive active restoration. SageSTEP is designed as a long‐term study, such that measurements are planned for at least 10 years after treatment implementation, or through the 2018 field season. This final report therefore describes the short‐term effects of treatments, 2‐4 years after treatment implementation., or through the 2010 field season. The Joint Fire Science Program generously funded SageSTEP for its first six years, and this funding was crucial for building an infrastructure that has now set the stage for an unprecedented long‐term study that will provide badly needed information on sagebrush steppe restoration and fuel treatment effectiveness. The infrastructure we’ve built consists of the following eight features: 1. A network of 18 sites distributed across the Great Basin, Snake River Basin, and Columbia Basin, 11 sites in a replicated woodland experiment, and 7 sites in a replicated sage‐cheat experiment (Figure 1). Each site is equivalent to a statistical block consisting of an unmanipulated control, and a set of fire and fire surrogate treatments. 2. A network of weather and soil moisture stations distributed along with the sites, that provides information on inter‐annual and geographic variation in moisture and temperature, and that is being used to interpret patterns of ecological response. 3. A small by efficient staff, consisting of scientists and technicians, responsible for continued monitoring of ecological variables through time, and maintenance of the projects’ infrastructure. 4. A funding stream from several agency sources, with current resources adequate to run the project for at least three more years, and with agreements in place to fund the project through fiscal year 2015. 5. A web of partnerships among managers, scientists, students, stakeholders, and policymakers that has worked together to design the study, implement the treatments, and learn about how sagebrush steppe system respond to alternative restoration treatments. 6. A highly effective and influential outreach program, anchored by a popular website, designed to interpret and deliver scientific information collected by SageSTEP scientists, and to distribute other relevant information originating from outside the project. 7. An on‐line database, called the SageSTEP Data Store, that offers fully proofed and validated data to analysts working within SageSTEP, and which will eventually provide the same information to other interested users. 8. The Great Basin NEON Site, NSF’s atmospheric sampling station that will soon be built at the SageSTEP Onaqui site. This link with NSF provides SageSTEP with leverage for established additional vegetation and soil monitoring facilities at Onaqui. Over the past three years, since post‐treatment data collection commenced, SageSTEP has produced a considerable amount of information, most of it now published in a total of 32 scientific papers. Key outreach products include: ● Active web site (sagestep.org), anchoring a comprehensive outreach program ● User\u27s Guides for Western Juniper & Pinyon‐Juniper woodlands ● Two Fuel Guides, one each for pre‐treatment and post‐treatment conditions ● 15 quarterly newsletters ● Six manager workshops ● 11 tours or field trips ● Three national conference symposia, consisting of 24 papers (2 symposia planned) ● 57 contributed papers at conferences ● Seven Master’s Theses and two Ph.D. Dissertations ● 15 papers published in proceedings or reports ● Ten papers published in peer‐reviewed journals (17 papers currently in review

Hyperstable Synthetic Mini-Proteins as Effective Ligand Scaffolds

Small, single-domain protein scaffolds are compelling sources of molecular binding ligands with the potential for efficient physiological transport, modularity, and manufacturing. Yet, mini-proteins require a balance between biophysical robustness and diversity to enable new functions. We tested the developability and evolvability of millions of variants of 43 designed libraries of synthetic 40-amino acid βαββ proteins with diversified sheet, loop, or helix paratopes. We discovered a scaffold library that yielded hundreds of binders to seven targets while exhibiting high stability and soluble expression. Binder discovery yielded 6–122 nM affinities without affinity maturation and Tms averaging ≥78 °C. Broader βαββ libraries exhibited varied developability and evolvability. Sheet paratopes were the most consistently developable, and framework 1 was the most evolvable. Paratope evolvability was dependent on target, though several libraries were evolvable across many targets while exhibiting high stability and soluble expression. Select βαββ proteins are strong starting points for engineering performant binders

A REGIONAL EXPERIMENT TO EVALUATE EFFECTS OF FIRE AND FIRE SURROGATE TREATMENTS IN THE SAGEBRUSH BIOME

SageSTEP is a comprehensive regional experiment that provides critical information to managers faced with a sagebrush steppe ecosystem that is increasingly at risk from wildfire, invasive plants, and climate change. The experiment provides managers with information that can be used to restore ecological communities across the 100+ million acres of the sagebrush biome. It is designed to match the temporal and spatial scales at which managers operate, is intended to reduce management risk and uncertainty of catastrophic wildfire to the greatest degree possible, and provides managers with information that allows them to better understand tradeoffs inherent in the choice of management alternatives. The project has several features that make it ideal for testing hypotheses from state‐andtransition theory, and for discovering information that can be directly applied in a management context ‐‐ it is long‐term, experimental, multisite, multivariate, and treatments are applied across condition gradients, allowing for potential identification of biotic thresholds. The project is designed to distinguish communities that have conditions that will allow them to recover on their own following fuel or restoration treatments, versus communities that have crossed biotic thresholds, and will therefore require more expensive active restoration. SageSTEP is designed as a long‐term study, such that measurements are planned for at least 10 years after treatment implementation, or through the 2018 field season. This final report therefore describes the short‐term effects of treatments, 2‐4 years after treatment implementation., or through the 2010 field season. The Joint Fire Science Program generously funded SageSTEP for its first six years, and this funding was crucial for building an infrastructure that has now set the stage for an unprecedented long‐term study that will provide badly needed information on sagebrush steppe restoration and fuel treatment effectiveness. The infrastructure we’ve built consists of the following eight features: 1. A network of 18 sites distributed across the Great Basin, Snake River Basin, and Columbia Basin, 11 sites in a replicated woodland experiment, and 7 sites in a replicated sage‐cheat experiment (Figure 1). Each site is equivalent to a statistical block consisting of an unmanipulated control, and a set of fire and fire surrogate treatments. 2. A network of weather and soil moisture stations distributed along with the sites, that provides information on inter‐annual and geographic variation in moisture and temperature, and that is being used to interpret patterns of ecological response. 3. A small by efficient staff, consisting of scientists and technicians, responsible for continued monitoring of ecological variables through time, and maintenance of the projects’ infrastructure. 4. A funding stream from several agency sources, with current resources adequate to run the project for at least three more years, and with agreements in place to fund the project through fiscal year 2015. 5. A web of partnerships among managers, scientists, students, stakeholders, and policymakers that has worked together to design the study, implement the treatments, and learn about how sagebrush steppe system respond to alternative restoration treatments. 6. A highly effective and influential outreach program, anchored by a popular website, designed to interpret and deliver scientific information collected by SageSTEP scientists, and to distribute other relevant information originating from outside the project. 7. An on‐line database, called the SageSTEP Data Store, that offers fully proofed and validated data to analysts working within SageSTEP, and which will eventually provide the same information to other interested users. 8. The Great Basin NEON Site, NSF’s atmospheric sampling station that will soon be built at the SageSTEP Onaqui site. This link with NSF provides SageSTEP with leverage for established additional vegetation and soil monitoring facilities at Onaqui. Over the past three years, since post‐treatment data collection commenced, SageSTEP has produced a considerable amount of information, most of it now published in a total of 32 scientific papers. Key outreach products include: ● Active web site (sagestep.org), anchoring a comprehensive outreach program ● User\u27s Guides for Western Juniper & Pinyon‐Juniper woodlands ● Two Fuel Guides, one each for pre‐treatment and post‐treatment conditions ● 15 quarterly newsletters ● Six manager workshops ● 11 tours or field trips ● Three national conference symposia, consisting of 24 papers (2 symposia planned) ● 57 contributed papers at conferences ● Seven Master’s Theses and two Ph.D. Dissertations ● 15 papers published in proceedings or reports ● Ten papers published in peer‐reviewed journals (17 papers currently in review

Coordinated Action of Two Double-Stranded RNA Binding Motifs and an RGG Motif Enables Nuclear Factor 90 To Flexibly Target Different RNA Substrates

The mechanisms of how RNA binding proteins (RBP) bind to and distinguish different RNA molecules are yet uncertain. Here, we performed a comprehensive analysis of the RNA binding properties of multidomain RBP nuclear factor 90 (NF90) by investigating specifically the functional activities of two double-stranded RNA binding motifs (dsRBM) and an RGG motif in the protein’s unstructured C-terminus. By comparison of the RNA binding affinities of several NF90 variants and their modes of binding to a set of defined RNA molecules, the activities of the motifs turned out to be very different. While dsRBM1 contributes little to RNA binding, dsRBM2 is essential for effective binding of double-stranded RNA. The protein’s immediate C-terminus, including the RGG motif, is indispensable for interactions of the protein with single-stranded RNA, and the RGG motif decisively contributes to NF90’s overall RNA binding properties. Conformational studies, which compared wild-type NF90 with a variant that contains a pseudophosphorylated residue in the RGG motif, suggest that the NF90 C-terminus is involved in conformational changes in the protein after RNA binding, with the RGG motif acting as a central regulatory element. In summary, our data propose a concerted action of all RNA binding motifs within the frame of the full-length protein, which may be controlled by regulation of the activity of the RGG motif, e.g., by phosphorylation. This multidomain interplay enables the RBP NF90 to discriminate RNA features by dynamic and adaptable interactions

Initiation of RNA Synthesis by the Hepatitis C Virus RNA-Dependent RNA Polymerase Is Affected by the Structure of the RNA Template

The hepatitis C virus (HCV) RNA-dependent RNA polymerase NS5B is a central enzyme of the intracellular replication of the viral (+)­RNA genome. Here, we studied the individual steps of NS5B-catalyzed RNA synthesis by a combination of biophysical methods, including real-time 1D ¹H NMR spectroscopy. NS5B was found to bind to a nonstructured and a structured RNA template in different modes. Following NTP binding and conversion to the catalysis-competent ternary complex, the polymerase revealed an improved affinity for the template. By monitoring the folding/unfolding of 3′(−)­SL by ¹H NMR, the base pair at the stem’s edge was identified as the most stable component of the structure. ¹H NMR real-time analysis of NS5B-catalyzed RNA synthesis on 3′(−)­SL showed that a pronounced lag phase preceded the processive polymerization reaction. The presence of the double-stranded stem with the edge base pair acting as the main energy barrier impaired RNA synthesis catalyzed by NS5B. Our observations suggest a crucial role of RNA-modulating factors in the HCV replication process