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Genetic improvement through breeding is one of the key approaches to increasing biomass supply. This paper documents the breeding progress to date for four perennial biomass crops (PBCs) that have high output–input energy ratios: namely Panicum virgatum (switchgrass), species of the genera Miscanthus (miscanthus), Salix (willow) and Populus (poplar). For each crop, we report on the size of germplasm collections, the efforts to date to phenotype and genotype, the diversity available for breeding and on the scale of breeding work as indicated by number of attempted crosses. We also report on the development of faster and more precise breeding using molecular breeding techniques. Poplar is the model tree for genetic studies and is furthest ahead in terms of biological knowledge and genetic resources. Linkage maps, transgenesis and genome editing methods are now being used in commercially focused poplar breeding. These are in development in switchgrass, miscanthus and willow generating large genetic and phenotypic data sets requiring concomitant efforts in informatics to create summaries that can be accessed and used by practical breeders. Cultivars of switchgrass and miscanthus can be seed‐based synthetic populations, semihybrids or clones. Willow and poplar cultivars are commercially deployed as clones. At local and regional level, the most advanced cultivars in each crop are at technology readiness levels which could be scaled to planting rates of thousands of hectares per year in about 5 years with existing commercial developers. Investment in further development of better cultivars is subject to current market failure and the long breeding cycles. We conclude that sustained public investment in breeding plays a key role in delivering future mass‐scale deployment of PBCs. Plant breeding links the research effort with commercial mass upscaling. The authors’ assessment of development status of the four species is shown (poplar having two: one for short rotation coppice (SRC) poplar and one for the more traditional short rotation forestry (SRF)). Mass scale deployment needs developments outside the breeding arenas to drive breeding activities more rapidly and extensively.

Hydraulic and chemical signals operate in tandem to regulate systemic plant responses to drought. Transport of abscisic acid (ABA) through the xylem and phloem from the root to shoot has been suggested to serve as the main signal of water deficit. There is evidence that ABA and its ABA-glycosyl-ester (ABA-GE) are also formed in leaves and stems through the chloroplastic 2-C-methylerythritol-5-phosphate (MEP) pathway. This study aimed to evaluate how hormonal and hydraulic signals contribute to optimize stomatal (gs), mesophyll (gm) and leaf hydraulic (Kleaf) conductance under well-watered and water-stressed conditions in Populus nigra (black poplar) plants. In addition, we assessed possible relationships between ABA and soluble carbohydrates within the leaf and stem. Plants were subjected to three water treatments: well-watered (WW), moderate stress (WS1) and severe stress (WS2). This experimental set-up enabled a time-course analysis of the response to water deficit at the physiological [leaf gas exchange, plant water relations, (Kleaf)], biochemical (ABA and its metabolite/catabolite quantification in xylem sap, leaves, wood, bark and roots) and molecular (gene expression of ABA biosynthesis) levels. Our results showed strong coordination between gs, gm and Kleaf under water stress, which reduced transpiration and increased intrinsic water use efficiency (WUEint). Analysis of gene expression of 9-cis-epoxycarotenoid dioxygenase (NCED) and ABA content in different tissues showed a general up-regulation of the biosynthesis of this hormone and its finely-tuned catabolism in response to water stress. Significant linear relationships were found between soluble carbohydrates and ABA contents in both leaves and stems, suggesting a putative function for this hormone in carbohydrate mobilization under severe water stress. This study demonstrates the tight regulation of the photosynthetic machinery by levels of ABA in different plants organs on a daily basis in both well-watered and water stress conditions to optimize WUEint and coordinate whole plant acclimation responses to drought.

The advent of molecular breeding is advocated to improve the productivity and sustainability of second-generation bioenergy crops. Advanced molecular breeding in bioenergy crops relies on the ability to massively sample the genetic diversity. Genotyping-by-sequencing has become a widely adopted method for cost-effective genotyping. It basically requires no initial investment for design as compared with array-based platforms which have been shown to offer very robust assays. The latter, however, has the drawback of being limited to analyse only the genetic diversity accounted during selection of a set of polymorphisms and design of the assay. In contrast, genotyping-by-sequencing with random sampling of genomic loci via restriction enzymes or random priming has been shown to be fast and convenient but lacks the ability to target specific regions of the genome and to maintain high reproducibility across laboratories. Here we present a first adoption of single-primer enrichment technology (SPET) which provides a highly efficient and scalable system to obtain targeted sequence-based large genotyping data sets, bridging the gaps between array-based systems and traditional sequencing-based protocols. To fully explore SPET performance, we conducted a benchmark study in ten Zea mays lines and a large-scale study of a natural black poplar population of 540 individuals with the aim of discovering polymorphisms associated with biomass-related traits. Our results showed the ability of this technology to provide dense genotype information on a customized panel of selected polymorphisms, while yielding hundreds of thousands of untargeted variable sites. This provided an ideal resource for association analysis of natural populations harbouring unexplored allelic diversities and structure such as in black poplar. The improvement of sequencing throughput and the development of efficient library preparation methods has made it feasible to carry out targeted genotyping-by-sequencing experiments cost-competitively with either random complexity reduction systems or traditional array-based platforms, while maintaining the key advantages of both technologies.

Perennial bioenergy crops accumulate carbon (C) in soils through minimally disturbing management practices and large root inputs, but the mechanisms of microbial control over C dynamics under bioenergy crops have not been clarified. Root‐derived C inputs affect both soil microbial contribution to and degradation of soil organic matter resulting in differing soil organic carbon (SOC) concentrations, storage, and stabilities under different vegetation regimes. Here, we measured biomarker amino sugars and neutral sugars and used diffuse reflectance mid‐infrared Fourier transform spectroscopy (DRIFTS) to explore microbial C contributions, degradation ability, and SOC stability, respectively, under four potential bioenergy crops, M.×giganteus (Miscanthus × giganteus), switchgrass (Panicum virgatum L.), a mixed prairie, and a maize (Zea mays L.)–maize–soybean (Glycine max(L.) Merr.) (MMS) rotation over six growing seasons. Our results showed that SOC concentration (g/kg) increased by 10.6% in mixed prairie over the duration of this experiment and SOC storage (Mg/ha) increased by 17.0% and 15.6% in switchgrass and mixed prairie, respectively. Conversion of row crops to perennial grasses maintained SOC stability and increased bacterial residue contribution to SOC in M.×giganteus and switchgrass by 20.0% and 15.0%, respectively, after 6 years. Degradation of microbe‐derived labile SOC was increased in M.×giganteus, and degradation of both labile and stable SOC increased in MMS rotation. These results demonstrate that microbial communities under perennial grasses maintained SOC quality, while SOC quantity increased under switchgrass and mixed prairie. Annual MMS rotation displayed decreases in aspects of SOC quality without changes in SOC quantity. These findings have implications for understanding microbial control over soil C quantity and quality under land‐use shift from annual to perennial bioenergy cropping systems. Microbial substrate preference is speculated to be driven by labile carbon inputs and available nitrogen. The results from this study demonstrate optimal conditions for increasing soil organic carbon quantity and quality beyond a cessation of tillage include a diverse aboveground ecosystem, high belowground labile carbon inputs, and available nitrogen.

Miscanthus, a high-yielding, warm-season C4 grass, shows promise as a potential bioenergy crop in temperate regions. However, drought may restrain productivity of most genotypes. In this study, total 29 Miscanthus genotypes of East-Asian origin were screened for drought tolerance with two methods, a dry-down treatment in two locations and a system where soil moisture content (SMC) was maintained at fixed levels using an automatic irrigation system in one location. One genotype, Miscanthus sinensis PMS-285, showed relatively high drought-tolerance capacity under moderate drought stress. Miscanthus sinensis PMS-285, aligned with the M. sinensis ‘Yangtze-Qinling’ genetic cluster, had relatively high principal component analysis ranking values in both two locations experiments, Hokkaido University and Brigham Young University. Genotypes derived from the ‘Yangtze-Qinling’ genetic cluster showed relatively greater photosynthetic performance than other genetic clusters, suggesting germplasm from this group could be a potential source of drought-tolerant plant material. Diploid genotypes showed stronger drought tolerance than tetraploid genotypes, suggesting ploidy could be an influential factor for this trait. Of the two methods, the dry-down treatment appears more suitable for selecting drought-tolerant genotypes given that it reflects water-stress conditions in the field. However, the fixed-SMC experiment may be good for understanding the physiological responses of plants to relatively constant water-stress levels.

Nitrogen is an essential element of life, and nitrogen availability often limits crop yields. Since the Green Revolution, massive amounts of synthetic nitrogen fertilizers have been produced from atmospheric nitrogen and natural gas, threatening the sustainability of global food production and degrading the environment. There is a need for alternative means of bringing nitrogen to crops, and taking greater advantage of biological nitrogen fixation seems a logical option. Legumes are used in most cropping systems around the world because of the nitrogen-fixing symbiosis with rhizobia. However, the world's three major cereal crops—rice, wheat, and maize—do not associate with rhizobia. In this review, we will survey how genetic approaches in rhizobia and their legume hosts allowed tremendous progress in understanding the molecular mechanisms controlling root nodule symbioses, and how this knowledge paves the way for engineering such associations in non-legume crops. We will also discuss challenges in bringing these systems into the field and how they can be surmounted by interdisciplinary collaborations between synthetic biologists, microbiologists, plant biologists, breeders, agronomists, and policymakers.

The need for renewable energy sources will lead to a considerable expansion in the planting of dedicated fast‐growing biomass crops across Europe. These are commonly cultivated as short‐rotation coppice (SRC), and currently poplar (Populus spp.) is the most widely planted. In this study, we report the greenhouse gas (GHG) fluxes of carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) measured using eddy covariance technique in an SRC plantation for bioenergy production. Measurements were made during the period 2010–2013, that is, during the first two rotations of the SRC. The overall GHG balance of the 4 years of the study was an emission of 1.90 (±1.37) Mg CO2eq ha−1; this indicated that soil trace gas emissions offset the CO2 uptake by the plantation. CH4 and N2O contributed almost equally to offset the CO2 uptake of −5.28 (±0.67) Mg CO2eq ha−1 with an overall emission of 3.56 (±0.35) Mg CO2eq ha−1 of N2O and of 3.53 (±0.85) Mg CO2eq ha−1 of CH4. N2O emissions mostly occurred during one single peak a few months after the site was converted to SRC; this peak comprised 44% of the total N2O loss during the two rotations. Accurately capturing emission events proved to be critical for deriving correct estimates of the GHG balance. The nitrogen (N) content of the soil and the water table depth were the two drivers that best explained the variability in N2O and CH4, respectively. This study underlines the importance of the ‘non‐CO2 GHGs’ on the overall balance. Further long‐term investigations of soil trace gas emissions should monitor the N content and the mineralization rate of the soil, as well as the microbial community, as drivers of the trace gas emissions.

Miscanthus is a perennial wild grass that is of global importance for paper production, roofing, horticultural plantings, and an emerging highly productive temperate biomass crop. We report a chromosome-scale assembly of the paleotetraploid M. sinensis genome, providing a resource for Miscanthus that links its chromosomes to the related diploid Sorghum and complex polyploid sugarcanes. The asymmetric distribution of transposons across the two homoeologous subgenomes proves Miscanthus paleo-allotetraploidy and identifies several balanced reciprocal homoeologous exchanges. Analysis of M. sinensis and M. sacchariflorus populations demonstrates extensive interspecific admixture and hybridization, and documents the origin of the highly productive triploid bioenergy crop M. × giganteus . Transcriptional profiling of leaves, stem, and rhizomes over growing seasons provides insight into rhizome development and nutrient recycling, processes critical for sustainable biomass accumulation in a perennial temperate grass. The Miscanthus genome expands the power of comparative genomics to understand traits of importance to Andropogoneae grasses. The perennial grass Miscanthus is a promising biomass crop. Here, via genomics and transcriptomics, the authors reveal its allotetraploid origin, characterize gene expression associated with rhizome development and nutrient recycling, and describe the hybrid origin of the triploid M. x giganteus .

Coproduction of high-value bioproducts at biorefineries is a key factor in making biofuels more cost-competitive. One strategy for generating coproducts is to directly engineer bioenergy crops to accumulate bioproducts in planta that can be fractionated and recovered at biorefineries. Here, we develop quantitative insights into the relationship between bioproduct market value and target accumulation rates by investigating a set of industrially relevant compounds already extracted from plant sources with a wide range of market prices and applications, including <$10/kg (limonene, latex, and polyhydroxybutyrate [PHB]), $10 to $100/kg (cannabidiol), and >$100/kg (artemisinin). These compounds are used to identify a range of mass fraction thresholds required to achieve net economic benefits for biorefineries and the additional amounts needed to reach a target $2.50/gal biofuel selling price, using cellulosic ethanol production as a test case. Bioproduct market prices and recovery costs determine the accumulation threshold; we find that moderate- to high-value compounds (i.e., cannabidiol and artemisinin) offer net economic benefits at accumulation rates of just 0.01% dry weight (dwt) to 0.02 dwt%. Lower-value compounds, including limonene, latex, and PHB, require at least an order-of-magnitude greater accumulation to overcome additional extraction and recovery costs (0.3 to 1.2 dwt%). We also find that a diversified approach is critical. For example, global artemisinin demand could be met with fewer than 10 biorefineries, while global demand for latex is equivalent to nearly 180 facilities. Our results provide a roadmap for future plant metabolic engineering efforts aimed at increasing the value derived from bioenergy crops.

The growing bioeconomy will require a greater supply of biomass in the future for both bioenergy and bio-based products. Today, many bioenergy cropping systems (BCS) are suboptimal due to either social-ecological threats or technical limitations. In addition, the competition for land between bioenergy-crop cultivation, food-crop cultivation, and biodiversity conservation is expected to increase as a result of both continuous world population growth and expected severe climate change effects. This study investigates how BCS can become more social-ecologically sustainable in future. It brings together expert opinions from the fields of agronomy, economics, meteorology, and geography. Potential solutions to the following five main requirements for a more holistically sustainable supply of biomass are summarized: (i) bioenergy-crop cultivation should provide a beneficial social-ecological contribution, such as an increase in both biodiversity and landscape aesthetics, (ii) bioenergy crops should be cultivated on marginal agricultural land so as not to compete with food-crop production, (iii) BCS need to be resilient in the face of projected severe climate change effects, (iv) BCS should foster rural development and support the vast number of small-scale family farmers, managing about 80% of agricultural land and natural resources globally, and (v) bioenergy-crop cultivation must be planned and implemented systematically, using holistic approaches. Further research activities and policy incentives should not only consider the economic potential of bioenergy-crop cultivation, but also aspects of biodiversity, soil fertility, and climate change adaptation specific to site conditions and the given social context. This will help to adapt existing agricultural systems in a changing world and foster the development of a more social-ecologically sustainable bioeconomy.