DArT-based characterisation of genetic diversity in a Miscanthus collection from Poland.

MAIN CONCLUSION: Analysis of 180 accessions of Miscanthus using a DArT platform revealed high diversity. The phylogenetic analysis revealed that M. × giganteus accessions fall into two genetically distinct groups. Miscanthus is a genus of perennial rhizomatous grasses that has emerged in last 20 years as a feedstock for bioenergy and biofuel production. Currently, the most widely used accession for bioenergy purposes is Miscanthus × giganteus, a sterile triploid hybrid between Miscanthus sinensis and Miscanthus sacchariflorus. However, previous reports have shown that genetic diversity of Miscanthus × giganteus is limited. Here, we report development of Diversity Arrays Technology platform for the analysis of genetic structure of a Miscanthus collection of 180 accessions. A total of 906 markers were obtained of which around 25.5% exhibited polymorphism information content value in the range of 0.40 and 0.50 and are considered particularly informative. Newly developed marker system will serve as an additional resource to assist crop improvement, germplasm preservation and genetic studies. Three types of analysis indicated that 180 accessions from the collection were well differentiated and presented high diversity. Interestingly, the analysis revealed that there are two separate groups of plants, significantly differing in genetic diversity, that are commercially available as M. × giganteus. We suggest that one of these groups is most likely mutants or somaclonal variants of original M. × giganteus. The other group is recent hybrids of Miscanthus of higher genetic diversity. This study indicates that the diversity of commercially available M. × giganteus is higher than commonly assumed. Development of the new marker system can significantly assist breeding of new commercial cultivars of Miscanthus for bioenergy use.

Improving field establishment and yield in seed propagated Miscanthus through manipulating plug size, sowing date and seedling age.

Biomass crops provide significant potential to substitute for fossil fuels and mitigate against climate change. It is widely acknowledged that significant scale up of biomass crops is required to help reach net zero targets. Miscanthus is a leading biomass crop embodying many characteristics that make it a highly sustainable source of biomass but planted area remains low. Miscanthus is commonly propagated via rhizome, but efficient alternatives may increase uptake and help diversify the cultivated crop. Using seed-propagate plug plants of Miscanthus has several potential benefits such as improving propagation rates and scale up of plantations. Plugs also provide an opportunity to vary the time and conditions under protected growth, to achieve optimal plantlets before planting. We varied combinations of glasshouse growth period and field planting dates under UK temperate conditions, which demonstrated the special importance of planting date on yield, stem number and establishment rates of Miscanthus. We also propagated Miscanthus in four different commercial plug designs that contained different volumes of substrate, the resulting seedlings were planted at three different dates into field trials. In the glasshouse, plug design had significant effects on above and belowground biomass accumulation and at a later time point belowground growth was restricted in some plug designs. After subsequent growth in the field, plug design and planting date had a significant effect on yield. The effects of plug design on yield were no longer significant after a second growth season but planting date continued to have a significant effect. After the second growth year, it was found that planting date had a significant effect on surviving plants, with the mid-season planting producing higher survival rates over all plug types.Establishment was positively correlated with DM biomass produced in the first growth season. Sowing date had a significant effect on establishment but the impacts of plug design were more nuanced and were significant at later planting dates. We discuss the potential to use the flexibility afforded by seed propagation of plug plants to deliver significant impacts in achieving high yield and establishment of biomass crops during the critical first two years of growth.

Perennial biomass cropping and use: Shaping the policy ecosystem in European countries.

Demand for sustainably produced biomass is expected to increase with the need to provide renewable commodities, improve resource security and reduce greenhouse gas emissions in line with COP26 commitments. Studies have demonstrated additional environmental benefits of using perennial biomass crops (PBCs), when produced appropriately, as a feedstock for the growing bioeconomy, including utilisation for bioenergy (with or without carbon capture and storage). PBCs can potentially contribute to Common Agricultural Policy (CAP) (2023-27) objectives provided they are carefully integrated into farming systems and landscapes. Despite significant research and development (R&D) investment over decades in herbaceous and coppiced woody PBCs, deployment has largely stagnated due to social, economic and policy uncertainties. This paper identifies the challenges in creating policies that are acceptable to all actors. Development will need to be informed by measurement, reporting and verification (MRV) of greenhouse gas emissions reductions and other environmental, economic and social metrics. It discusses interlinked issues that must be considered in the expansion of PBC production: (i) available land; (ii) yield potential; (iii) integration into farming systems; (iv) R&D requirements; (v) utilisation options; and (vi) market systems and the socio-economic environment. It makes policy recommendations that would enable greater PBC deployment: (1) incentivise farmers and land managers through specific policy measures, including carbon pricing, to allocate their less productive and less profitable land for uses which deliver demonstrable greenhouse gas reductions; (2) enable greenhouse gas mitigation markets to develop and offer secure contracts for commercial developers of verifiable low-carbon bioenergy and bioproducts; (3) support innovation in biomass utilisation value chains; and (4) continue long-term, strategic R&D and education for positive environmental, economic and social sustainability impacts.

New Miscanthus hybrids cultivated at a Polish metal-contaminated site demonstrate high stomatal regulation and reduced shoot Pb and Cd concentrations.

The increased bioeconomy targets for the biomass share of renewable energy production across Europe should be met using land unsuitable for food production. Miscanthus breeding programs targeted the production of plants with a diverse range of traits allowing a wider utilization of land resources for biofuel production without competing with arable crops. These traits include increasing tolerances to drought, chilling, and to metal(loid)s excess. Two novel Miscanthus hybrids, GNT41 and GNT34, were compared against Miscanthus x giganteus (Mxg) on metal-contaminated arable land in Poland. This study aimed at evaluating their yield, biomass quality and quantifying seasonal differences in photosynthetic and transpiration parameters. A secondary objective was to identify key physiological mechanisms underlying differences in metal accumulation between the investigated plants. The new hybrids produced a similar yield to Mxg (13-15 t ha-1 yr-1), had shorter shoots, higher Leaf Area Index and stem number. Based on gas exchange measurements, GNT34 exhibited isohydric (water-conserving) behavior. The stomatal response to light of the new hybrids was at least twice as fast as that of Mxg, a trait that is often associated with increased seasonal water use efficiency. This contributed to the almost 40% reduction in shoot Pb and Cd concentrations for the new hybrids as compared to Mxg. This suggested that promoting stomatal regulation in conjunction with improved water conservation may be a target for improving plants for wider use on metals contaminated land.

Breeding progress and preparedness for mass-scale deployment of perennial lignocellulosic biomass crops switchgrass, miscanthus, willow and poplar.

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.

Assessing seed priming, sowing date, and mulch film to improve the germination and survival of direct-sown Miscanthus sinensis in the United Kingdom.

Direct sowing of Miscanthus seed could lower crop establishment costs, and increase the rate of grower uptake and biomass supply for the emerging bio-economy. A replicated field trial was conducted at two contrasting UK sites: Aberystwyth (ABR) in mid-Wales and Blankney (BLK) in Lincolnshire. These sites encompass the west-east meteorological gradient in the United Kingdom where the growing season at ABR is cooler and wetter while BLK is warmer and drier. Primed and unprimed Miscanthus sinensis seeds were sown directly onto the soil surface with and without a clear biodegradable mulch film, at nine dates interspersed from May to October. Average daily mean soil surface temperatures measured over the first 2 months after sowing under the mulch film were higher than control plots (2.7°C ABR and 4.2°C BLK). At both sites, the film covering also affected soil volumetric moisture relative to uncovered control plots (-3% ABR and 8% BLK), demonstrating the negative impact of mulch film when sowing on dry soil. Over nine sowings, seed germination at ABR under film varied between -28% and +18% of germination under control conditions. Seedlings from the first three sowings at both sites under film had sufficient physiological maturity to survive the first winter period. At BLK, mulch film significantly increased tiller count and height in both the first and second years after sowing. At ABR, where temperatures were lower, film covering significantly increased tiller height but not count. Water priming had no significant effect on seed viability or germination in the field tests. Base temperatures for germination of primed and unprimed seeds on a thermal gradient plate were 7.0°C and 5.7°C, respectively, with a ± 1.7°C confidence interval. Based on our results for M. sinensis in the United Kingdom, we recommend the sowing of unprimed seed in May under film and only when the soil is moist.

Establishment, Growth, and Yield Potential of the Perennial Grass Miscanthus × Giganteus on Degraded Coal Mine Soils.

Miscanthus × giganteus is a giant C4 grass native to Asia. Unlike most C4 species, it is relatively cold tolerant due to adaptations across a wide range of altitudes. These grasses are characterized by high productivity and low input requirements, making them excellent candidates for bioenergy feedstock production. The aim of this study was to investigate the potential for growing Miscanthus on extremely marginal soils, degraded by open lignite (brown coal) mining. Field experiments were established within three blocks situated on waste heaps originating from the lignite mine. Analyses were conducted over the first 3 years following Miscanthus cultivation, focusing on the effect of organic and mineral fertilization on crop growth, development and yield in this extreme environment. The following levels of fertilization were implemented between the blocks: the control plot with no fertilization (D0), a plot with sewage sludge (D1), a plot with an identical amount of sewage sludge plus one dose of mineral fertilizer (D2) and a plot with an identical amount of sewage sludge plus a double dose of mineral fertilizer (D3). Crop development and characteristics (plant height, tillering, and biomass yield [dry matter]) were measured throughout the study period and analyzed using Analysis of Variance (ANOVA). Significant differences were apparent between plant development and 3rd year biomass production over the course of the study (0.964 kg plant-1 for DO compared to 1.503 kg plant-1 for D1). Soil analyses conducted over the course of the experiment showed that organic carbon levels within the soil increased significantly following the cultivation of Miscanthus, and overall, pH decreased. With the exception of iron, macronutrient concentrations remained stable throughout. The promising yields and positive effects of Miscanthus on the degraded soil suggests that long term plantations on land otherwise unsuitable for agriculture may prove to be of great environmental and economic significance.

Relationships between soil parameters and physiological status of Miscanthus x giganteus cultivated on soil contaminated with trace elements under NPK fertilisation vs. microbial inoculation.

Crop growth and development can be influenced by a range of parameters, soil health, cultivation and nutrient status all play a major role. Nutrient status of plants can be enhanced both through chemical fertiliser additions (e.g. N, P, K supplementation) or microbial fixation and mobilisation of naturally occurring nutrients. With current EU priorities discouraging the production of biomass on high quality soils there is a need to investigate the potential of more marginal soils to produce these feedstocks and the impacts of soil amendments on crop yields within them. This study investigated the potential for Miscanthus x giganteus to be grown in trace element (TE)-contaminated soils, ideally offering a mechanism to (phyto)manage these contaminated lands. Comprehensive surveys are needed to understand plant-soil interactions under these conditions. Here we studied the impacts of two fertiliser treatments on soil physico-chemical properties under Miscanthus x giganteus cultivated on Pb, Cd and Zn contaminated arable land. Results covered a range of parameters, including soil rhizosphere activity, arbuscular mycorrhization (AM), as well as plant physiological parameters associated with photosynthesis, TE leaf concentrations and growth performance. Fertilization increased growth and gas exchange capacity, enhanced rhizosphere microbial activity and increased Zn, Mg and N leaf concentration. Fertilization reduced root colonisation by AMF and caused higher chlorophyll concentration in plant leaves. Microbial inoculation seems to be a promising alternative for chemical fertilizers, especially due to an insignificant influence on the mobility of toxic trace elements (particularly Cd and Zn).

Economic and Environmental Assessment of Seed and Rhizome Propagated Miscanthus in the UK.

Growth in planted areas of Miscanthus for biomass in Europe has stagnated since 2010 due to technical challenges, economic barriers and environmental concerns. These limitations need to be overcome before biomass production from Miscanthus can expand to several million hectares. In this paper, we consider the economic and environmental effects of introducing seed based hybrids as an alternative to clonal M. x giganteus (Mxg). The impact of seed based propagation and novel agronomy was compared with current Mxg cultivation and used in 10 commercially relevant, field scale experiments planted between 2012 and 2014 in the United Kingdom, Germany, and Ukraine. Economic and greenhouse gas (GHG) emissions costs were quantified for the following production chain: propagation, establishment, harvest, transportation, storage, and fuel preparation (excluding soil carbon changes). The production and utilization efficiency of seed and rhizome propagation were compared. Results show that new hybrid seed propagation significantly reduces establishment cost to below £900 ha-1. Calculated GHG emission costs for the seeds established via plugs, though relatively small, was higher than rhizomes because fossil fuels were assumed to heat glasshouses for raising seedling plugs (5.3 and 1.5 kg CO2 eq. C Mg [dry matter (DM)]-1), respectively. Plastic mulch film reduced establishment time, improving crop economics. The breakeven yield was calculated to be 6 Mg DM ha-1 y-1, which is about half average United Kingdom yield for Mxg; with newer seeded hybrids reaching 16 Mg DM ha-1 in second year United Kingdom trials. These combined improvements will significantly increase crop profitability. The trade-offs between costs of production for the preparation of different feedstock formats show that bales are the best option for direct firing with the lowest transport costs (£0.04 Mg-1 km-1) and easy on-farm storage. However, if pelleted fuel is required then chip harvesting is more economic. We show how current seed based propagation methods can increase the rate at which Miscanthus can be scaled up; ∼×100 those of current rhizome propagation. These rapid ramp rates for biomass production are required to deliver a scalable and economic Miscanthus biomass fuel whose GHG emissions are ∼1/20th those of natural gas per unit of heat.

Extending Miscanthus Cultivation with Novel Germplasm at Six Contrasting Sites.

Miscanthus is a genus of perennial rhizomatous grasses with C4 photosynthesis which is indigenous in a wide geographic range of Asian climates. The sterile clone, Miscanthus × giganteus (M. × giganteus), is a naturally occurring interspecific hybrid that has been used commercially in Europe for biomass production for over a decade. Although, M. × giganteus has many outstanding performance characteristics including high yields and low nutrient offtakes, commercial expansion is limited by cloning rates, slow establishment to a mature yield, frost, and drought resistance. In this paper, we evaluate the performance of 13 novel germplasm types alongside M. × giganteus and horticultural "Goliath" in trials in six sites (in Germany, Russia, The Netherlands, Turkey, UK, and Ukraine). Mean annual yields across all the sites and genotypes increased from 2.3 ± 0.2 t dry matter ha-1 following the first year of growth, to 7.3 ± 0.3, 9.5 ± 0.3, and 10.5 ± 0.2 t dry matter ha-1 following the second, third, and fourth years, respectively. The highest average annual yields across locations and four growth seasons were observed for M. × giganteus (9.9 ± 0.7 t dry matter ha-1) and interspecies hybrid OPM-6 (9.4 ± 0.6 t dry matter ha-1). The best of the new hybrid genotypes yielded similarly to M. × giganteus at most of the locations. Significant effects of the year of growth, location, species, genotype, and interplay between these factors have been observed demonstrating strong genotype × environment interactions. The highest yields were recorded in Ukraine. Time needed for the crop establishment varied depending on climate: in colder climates such as Russia the crop has not achieved its peak yield by the fourth year, whereas in the hot climate of Turkey and under irrigation the yields were already high in the first growing season. We have identified several alternatives to M. × giganteus which have provided stable yields across wide climatic ranges, mostly interspecies hybrids, and also Miscanthus genotypes providing high biomass yields at specific geographic locations. Seed-propagated interspecific and intraspecific hybrids, with high stable yields and cheaper reliable scalable establishment remain a key strategic objective for breeders.

Environmental Influences on the Growing Season Duration and Ripening of Diverse Miscanthus Germplasm Grown in Six Countries.

The development of models to predict yield potential and quality of a Miscanthus crop must consider climatic limitations and the duration of growing season. As a biomass crop, yield and quality are impacted by the timing of plant developmental transitions such as flowering and senescence. Growth models are available for the commercially grown clone Miscanthus x giganteus (Mxg), but breeding programs have been working to expand the germplasm available, including development of interspecies hybrids. The aim of this study was to assess the performance of diverse germplasm beyond the range of environments considered suitable for a Miscanthus crop to be grown. To achieve this, six field sites were planted as part of the EU OPTIMISC project in 2012 in a longitudinal gradient from West to East: Wales-Aberystwyth, Netherlands-Wageningen, Stuttgart-Germany, Ukraine-Potash, Turkey-Adana, and Russia-Moscow. Each field trial contained three replicated plots of the same 15 Miscanthus germplasm types. Through the 2014 growing season, phenotypic traits were measured to determine the timing of developmental stages key to ripening; the tradeoff between growth (yield) and quality (biomass ash and moisture content). The hottest site (Adana) showed an accelerated growing season, with emergence, flowering and senescence occurring before the other sites. However, the highest yields were produced at Potash, where emergence was delayed by frost and the growing season was shortest. Flowering triggers varied with species and only in Mxg was strongly linked to accumulated thermal time. Our results show that a prolonged growing season is not essential to achieve high yields if climatic conditions are favorable and in regions where the growing season is bordered by frost, delaying harvest can improve quality of the harvested biomass.

Progress on Optimizing Miscanthus Biomass Production for the European Bioeconomy: Results of the EU FP7 Project OPTIMISC.

This paper describes the complete findings of the EU-funded research project OPTIMISC, which investigated methods to optimize the production and use of miscanthus biomass. Miscanthus bioenergy and bioproduct chains were investigated by trialing 15 diverse germplasm types in a range of climatic and soil environments across central Europe, Ukraine, Russia, and China. The abiotic stress tolerances of a wider panel of 100 germplasm types to drought, salinity, and low temperatures were measured in the laboratory and a field trial in Belgium. A small selection of germplasm types was evaluated for performance in grasslands on marginal sites in Germany and the UK. The growth traits underlying biomass yield and quality were measured to improve regional estimates of feedstock availability. Several potential high-value bioproducts were identified. The combined results provide recommendations to policymakers, growers and industry. The major technical advances in miscanthus production achieved by OPTIMISC include: (1) demonstration that novel hybrids can out-yield the standard commercially grown genotype Miscanthus x giganteus; (2) characterization of the interactions of physiological growth responses with environmental variation within and between sites; (3) quantification of biomass-quality-relevant traits; (4) abiotic stress tolerances of miscanthus genotypes; (5) selections suitable for production on marginal land; (6) field establishment methods for seeds using plugs; (7) evaluation of harvesting methods; and (8) quantification of energy used in densification (pellet) technologies with a range of hybrids with differences in stem wall properties. End-user needs were addressed by demonstrating the potential of optimizing miscanthus biomass composition for the production of ethanol and biogas as well as for combustion. The costs and life-cycle assessment of seven miscanthus-based value chains, including small- and large-scale heat and power, ethanol, biogas, and insulation material production, revealed GHG-emission- and fossil-energy-saving potentials of up to 30.6 t CO2eq C ha-1y-1 and 429 GJ ha-1y-1, respectively. Transport distance was identified as an important cost factor. Negative carbon mitigation costs of -78€ t-1 CO2eq C were recorded for local biomass use. The OPTIMISC results demonstrate the potential of miscanthus as a crop for marginal sites and provide information and technologies for the commercial implementation of miscanthus-based value chains.

Biological potential of microalgae in China for biorefinery-based production of biofuels and high value compounds.

Microalgae abundance and diversity in China shows promise for identifying suitable strains for developing algal biorefinery. Numerous strains of microalgae have already been assessed as feedstocks for bioethanol and biodiesel production, but commercial scale algal biofuel production is yet to be demonstrated, most likely due to huge energy costs associated with algae cultivation, harvesting and processing. Biorefining, integrated processes for the conversion of biomass into a variety of products, can improve the prospects of microalgal biofuels by combining them with the production of high value co-products. Numerous microalgal strains in China have been identified as producers of various high value by-products with wide application in the medicine, food, and cosmetics industries. This paper reviews microalgae resources in China and their potential in producing liquid biofuels (bioethanol and biodiesel) and high value products in an integrated biorefinery approach. Implementation of a 'high value product first' principle should make the integrated process of fuels and chemicals production economically feasible and will ensure that public and private interest in the development of microalgal biotechnology is maintained.