Effects of several establishment modes of Miscanthus × giganteus and Miscanthus sinensis on yields and yield trends

Miscanthus is a C4 perennial grass originating from East Asia, the yields of which progressively increase in the first years of growth. Several species for bioenergy have been studied since the mid‐1980s in Europe, in particular (Miscanthus × giganteus [M. × giganteus]), due to its high yields. M. × giganteus is mainly cultivated in France and established from rhizomes. Our study aimed to assess, in field conditions, alternative establishment methods combined with an alternative species, Miscanthus sinensis (M. sinensis). We set up a multi‐environment experimental network. On each trial, we tested two treatments with M. × giganteus, established from rhizomes (G_r‐sd) and from plantlets obtained from rhizomes (G_p‐sd), and two treatments with M. sinensis seedlings transplanted in single (S_p‐sd) and double density (S_p‐dd). ANOVA was performed to compare establishment and regrowth rates across treatments, as well as yields across treatments and site‐years. A logistic model was used to describe yield trends and to compare the maximum yield reached and the rate of yield increase of both species. Results showed that miscanthus establishment from plantlets resulted in higher establishment (between 87% and 92%) and regrowth (between 91% and 94%) rates compared to establishment from rhizomes. Treatments with M. × giganteus obtained higher average yields across site‐years than those with M. sinensis, but more variable yields across site‐years. We showed a strong species effect on yields, yield components (shoot weight, shoot density and shoot number per plant) and light interception (through leaf area index). Lastly, to use M. sinensis established from transplanted plantlets as an alternative to M. × giganteus, research would be required on the breeding of M. sinensis sterile seeds to avoid risks of invasiveness.     

Biomass yield in a genetically diverse Miscanthus sinensis germplasm panel evaluated at five locations revealed individuals with exceptional potential

To breed improved biomass cultivars of Miscanthus ×giganteus, it will be necessary to select the highest‐yielding and best‐adapted genotypes of its parental species, Miscanthus sinensis and Miscanthus sacchariflorus. We phenotyped a diverse clonally propagated panel of 569 M. sinensis and nine natural diploid M. ×giganteus at one subtropical (Zhuji, China) and five temperate locations (Sapporo, Japan; Leamington, Ontario, Canada; Fort Collins, CO; Urbana, IL; and Chuncheon, Korea) for dry biomass yield and 14 yield‐component traits, in trials grown for 3 years. Notably, dry biomass yield of four Miscanthus accessions exceeded 80 Mg/ha in Zhuji, China, approaching the highest observed for any land plant. Additionally, six M. sinensis in Sapporo, Japan and one in Leamington, Canada also yielded more than the triploid M. ×giganteus ‘1993‐1780’ control, with values exceeding 20 Mg/ha. Diploid M. ×giganteus was the best‐yielding group at the northern sites. Genotype‐by‐environment interactions were modest among the five northern trial sites but large between Zhuji, and the northern sites. M. sinensis accessions typically yielded best at trial sites with latitudes similar to collection sites, although broad adaptation was observed for accessions from southern Japan. Genotypic heritabilities for third year yields ranged from 0.71 to 0.88 within locations. Compressed circumference was the best predictor of yield. These results establish a baseline of data for initiating selection to improve biomass yield of M. sinensis and M. ×giganteus in a diverse set of relevant geographies.     

Genome Size of Three <Emphasis Type="Italic">Miscanthus</Emphasis> Species

Environmental and economic factors have stimulated research in the area of bioenergy crops. While many plants have been identified as potential energy crops, one species in particular, Miscanthus x giganteus, appears to have the most promise. As researchers attempt to exploit and improve M. x giganteus, genome information is critical. In this study, the genome size of M. x giganteus and its two progenitor species were examined by flow cytometry and stomatal cell analyses. M. x giganteus was found to have genome size of 7.0 pg while Miscanthus sinensis and Miscanthus sacchariflorus were observed to have genome sizes of 5.5 and 4.5 pg respectively with stomatal size correlating with genome size. Upon computing the two tetraploid × diploid hybrids theoretical genome sizes, the data presented in this paper supports the hypothesis of the union of a 2x M. sacchariflorus and a 1x M. sinensis gamete for the formation of the allotriploid, M. x giganteus. Such genomic information provides basic knowledge that is important in M. x giganteus plant improvement.     

Characterization of Miscanthus cell wall polymers

Efficient utilization of lignocellulosic Miscanthus biomass for the production of biochemicals, such as ethanol, is challenging due to its recalcitrance, which is influenced by the individual plant cell wall polymers and their interactions. Lignocellulosic biomass composition differs depending on several factors, such as plant age, harvest date, organ type, and genotype. Here, four selected Miscanthus genotypes (Miscanthus sinensis, Miscanthus sacchariflorus, Miscanthus × giganteus, Miscanthus sinensis × Miscanthus sacchariflorus hybrid) were grown and harvested, separated into stems and leaves, and characterized for their non‐starch polysaccharide composition and structures, lignin contents and structures, and hydroxycinnamate profiles (monomers and ferulic acid dehydrodimers). Polysaccharides of all genotypes are mainly composed of cellulose and low‐substituted arabinoxylans. Ratios of hemicelluloses to cellulose were comparable, with the exception of Miscanthus sinensis that showed a higher hemicellulose/cellulose ratio. Lignin contents of Miscanthus stems were higher than those of Miscanthus leaves. Considering the same organs, the four genotypes did not differ in their Klason lignin contents, but Miscanthus × giganteus showed the highest acetylbromide soluble lignin content. Lignin polymers isolated from stems varied in their S/G ratios and linkage type distributions across genotypes. p‐Coumaric acid was the most abundant ester‐bound hydroxycinnamte monomer in all samples. Ferulic acid dehydrodimers were analyzed as cell wall cross‐links, with 8‐5‐coupled diferulic acid being the main dimer, followed by 8‐O‐4‐, and 5‐5‐diferulic acid. Contents of p‐coumaric acid, ferulic acid, and ferulic acid dimers varied depending on genotype and organ type. The largest amount of cell wall cross‐links was analyzed for Miscanthus sinensis.     

The impacts of four potential bioenergy crops on soil carbon dynamics as shown by biomarker analyses and DRIFT spectroscopy

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.     

Growth and fecundity of fertile Miscanthus × giganteus (“PowerCane”) compared to feral and ornamental Miscanthus sinensis in a common garden experiment: Implications for invasion

Perennial grasses are promising candidates for bioenergy crops, but species that can escape cultivation and establish self‐sustaining naturalized populations (feral) may have the potential to become invasive. Fertile Miscanthus × giganteus, known as “PowerCane,” is a new potential biofuel crop. Its parent species are ornamental, non‐native Miscanthus species that establish feral populations and are sometimes invasive in the USA. As a first step toward assessing the potential for “PowerCane” to become invasive, we documented its growth and fecundity relative to one of its parent species (Miscanthus sinensis) in competition with native and invasive grasses in common garden experiments located in Columbus, Ohio and Ames, Iowa, within the targeted range of biofuel cultivation. We conducted a 2‐year experiment to compare growth and reproduction among three Miscanthus biotypes—”PowerCane,” ornamental M. sinensis, and feral M. sinensis—at two locations. Single Miscanthus plants were subjected to competition with a native grass (Panicum virgatum), a weedy grass (Bromus inermis), or no competition. Response variables were aboveground biomass, number of shoots, basal area, and seed set. In Iowa, all Miscanthus plants died after the first winter, which was unusually cold, so no further results are reported from the Iowa site. In Ohio, we found significant differences among biotypes in growth and fecundity, as well as significant effects of competition. Interactions between these treatments were not significant. “PowerCane” performed as well or better than ornamental or feral M. sinensis in vegetative traits, but had much lower seed production, perhaps due to pollen limitation. In general, ornamental M. sinensis performed somewhat better than feral M. sinensis. Our findings suggest that feral populations of “PowerCane” could become established adjacent to biofuel production areas. Fertile Miscanthus × giganteus should be studied further to assess its potential to spread via seed production in large, sexually compatible populations.     

Measured and modelled effect of land‐use change from temperate grassland to Miscanthus on soil carbon stocks after 12 years

Soil organic carbon (SOC) is an important carbon pool susceptible to land‐use change (LUC). There are concerns that converting grasslands into the C₄ bioenergy crop Miscanthus (to meet demands for renewable energy) could negatively impact SOC, resulting in reductions of greenhouse gas mitigation benefits gained from using Miscanthus as a fuel. This work addresses these concerns by sampling soils (0–30 cm) from a site 12 years (T₁₂) after conversion from marginal agricultural grassland into Miscanthus x giganteus and four other novel Miscanthus hybrids. Soil samples were analysed for changes in below‐ground biomass, SOC and Miscanthus contribution to SOC (using a ¹³C natural abundance approach). Findings are compared to ECOSSE soil carbon model results (run for a LUC from grassland to Miscanthus scenario and continued grassland counterfactual), and wider implications are considered in the context of life cycle assessments based on the heating value of the dry matter (DM) feedstock. The mean T₁₂ SOC stock at the site was 8 (±1 standard error) Mg C/ha lower than baseline time zero stocks (T₀), with assessment of the five individual hybrids showing that while all had lower SOC stock than at T₀ the difference was only significant for a single hybrid. Over the longer term, new Miscanthus C₄ carbon replaces pre‐existing C₃ carbon, though not at a high enough rate to completely offset losses by the end of year 12. At the end of simulated crop lifetime (15 years), the difference in SOC stocks between the two scenarios was 4 Mg C/ha (5 g CO₂‐eq/MJ). Including modelled LUC‐induced SOC loss, along with carbon costs relating to soil nitrous oxide emissions, doubled the greenhouse gas intensity of Miscanthus to give a total global warming potential of 10 g CO₂‐eq/MJ (180 kg CO₂‐eq/Mg DM).     

Plant and soil effects on bacterial communities associated with Miscanthus × giganteus rhizosphere and rhizomes

Bacterial assemblages, especially diazotroph assemblages residing in the rhizomes and the rhizosphere soil of Miscanthus × giganteus, contribute to plant growth and nitrogen use efficiency. However, the composition of these microbial communities has not been adequately explored nor have the potential ecological drivers for these communities been sufficiently studied. This knowledge is needed for understanding and potentially improving M. × giganteus – microbe interactions, and further enhancing sustainability of M. × giganteus production. In this study, cultivated M. × giganteus from four sites in Illinois, Kentucky, Nebraska, and New Jersey were collected to examine the relative influences of soil conditions and plant compartments on assembly of the M. × giganteus‐associated microbiome. Automated ribosomal intergenic spacer (ARISA) and terminal restriction fragment length polymorphism (T‐RFLP) targeting the nifH gene were applied to examine the total bacterial communities and diazotroph assemblages that reside in the rhizomes and the rhizosphere. Distinct microbial assemblages were detected in the endophytic and rhizosphere compartments. Site soil conditions had strong correlation with both total bacterial and diazotroph assemblages, but in different ways. Nitrogen treatments showed no significant effect on the composition of diazotroph assemblages in most sites. Endophytic compartments of different M. × giganteus plants tended to harbor similar microbial communities across all sites, whereas the rhizosphere soil of different plant tended to harbor diverse microbial assemblages that were distinct among sites. These observations offer insight into better understanding of the associative interactions between M. × giganteus and diazotrophs, and how this relationship is influenced by agronomic and edaphic factors.     

Soil carbon stock impacts following reversion of Miscanthus × giganteus and short rotation coppice willow commercial plantations into arable cropping

There are posited links between the establishment of perennial bioenergy, such as short rotation coppice (SRC) willow and Miscanthus × giganteus, on low carbon soils and enhanced soil C sequestration. Sequestration provides additional climate mitigation, however, few studies have explored impacts on soil C stocks of bioenergy crop removal; thus, the permanence of any sequestered C is unclear. This uncertainty has led some authors to question the handling of soil C stocks with carbon accounting, for example, through life cycle assessments. Here, we provide additional data for this debate, reporting on the soil C impacts of the reversion (removal and return) to arable cropping of commercial SRC willow and Miscanthus across four sites in the UK, two for each bioenergy crop, with eight reversions nested within these sites. Using a paired‐site approach, soil C stocks (0–1 m) were compared between 3 and 7 years after bioenergy crop removal. Impacts on soil C stocks varied, ranging from an increase of 70.16 ± 10.81 Mg C/ha 7 years after reversion of SRC willow to a decrease of 33.38 ± 5.33 Mg C/ha 3 years after reversion of Miscanthus compared to paired arable land. The implications for carbon accounting will depend on the method used to allocate this stock change between current and past land use. However, with published life cycle assessment values for the lifetime C reduction provided by these crops ranging from 29.50 to 138.55 Mg C/ha, the magnitude of these changes in stock are significant. We discuss the potential underlying mechanisms driving variability in soil C stock change, including the age of bioenergy crop at removal, removal methods, and differences in the recalcitrant of the crop residues, and highlight the need to design management methods to limit negative outcomes.     

Biomass production and energy balance of Miscanthus over a period of 11 years: A case study in a large‐scale farm in Poland

Giant miscanthus (Miscanthus × giganteus Greef and Deuter) and Amur silver grass (Miscanthus sacchariflorus Maxim./Hack) are rhizomatous grasses with a C4 photosynthetic pathway that are widely cultivated as energy crops. For those species to be successfully used in bioenergy generation, their yields have to be maintained at a high level in the long term. The biomass yield (fresh and dry matter [DM] yield) and energy efficiency (energy inputs, energy output, energy gain, and energy efficiency ratio) of giant miscanthus and Amur silver grass were compared in a field experiment conducted in 2007–2017 in North‐Eastern Poland. Both species were characterized by high above‐ground biomass yields, and the productive performance of M. × giganteus was higher in comparison with M. sacchariflorus (15.5 vs. 9.3 Mg DM ha^?1 year^?1 averaged for 1–11 years of growth). In the first year of the experiment, the energy inputs associated with the production of M. × giganteus and M. sacchariflorus were determined at 70.5 and 71.5 GJ/ha, respectively, and rhizomes accounted for around 78%–79% of total energy inputs. In the remaining years of cultivation, the total energy inputs associated with the production of both perennial rhizomatous grasses reached 13.6–15.7 (M. × giganteus) and 16.9–17.5 GJ ha^?1 year^?1 (M. sacchariflorus). Beginning from the second year of cultivation, mineral fertilizers were the predominant energy inputs in the production of M. × giganteus (78%–86%) and M. sacchariflorus (80%–82%). In years 2–11, the energy gain of M. × giganteus reached 50 (year 2) and 264–350 GJ ha^?1 year^?1 (years 3–11), and its energy efficiency ratio was determined at 4.7 (year 2) and 18.6–23.3 (years 3–11). The energy gain and the energy efficiency ratio of M. sacchariflorus biomass in the corresponding periods were determined at 87–234 GJ ha^?1 year^?1 and 6.1–14.3, respectively. Both grasses are significant and environmentally compatible sources of bioenergy, and they can be regarded as potential energy crops for Central‐Eastern Europe.     

A global meta‐analysis of soil organic carbon response to corn stover removal

Corn (Zea mays L.) stover is a global resource used for livestock, fuel, and bioenergy feedstock, but excessive stover removal can decrease soil organic C (SOC) stocks and deteriorate soil health. Many site‐specific stover removal experiments report accrual rates and SOC stock effects, but a quantitative, global synthesis is needed to provide a scientific base for long‐term energy policy decisions. We used 409 data points from 74 stover harvest experiments conducted around the world for a meta‐analysis and meta‐regression to quantify removal rate, tillage, soil texture, and soil sampling depth effects on SOC. Changes were quantified by: (a) comparing final SOC stock differences after at least 3 years with and without stover removal and (b) calculating SOC accrual rates for both treatments. Stover removal generally reduced final SOC stocks by 8% in the upper 0–15 or 0–30 cm, compared to stover retained, irrespective of soil properties and tillage practices. A more sensitive meta‐regression analysis showed that retention increased SOC stocks within the 30–150 cm depth by another 5%. Compared to baseline values, stover retention increased average SOC stocks temporally at a rate of 0.41 Mg C ha^?1 year^?1 (statistically significant at p < 0.01 when averaged across all soil layers). Although SOC sequestration rates were lower with stover removal, with moderate (<50%) removal they can be positive, thus emphasizing the importance of site‐specific management. Our results also showed that tillage effects on SOC stocks were inconsistent due to the high variability in practices used among the experimental sites. Finally, we conclude that research and technological efforts should continue to be given high priority because of the importance in providing science‐based policy recommendations for long‐term global carbon management.     

Genome‐wide association and genomic prediction for biomass yield in a genetically diverse Miscanthus sinensis germplasm panel phenotyped at five locations in Asia and North America

To improve the efficiency of breeding of Miscanthus for biomass yield, there is a need to develop genomics‐assisted selection for this long‐lived perennial crop by relating genotype to phenotype and breeding value across a broad range of environments. We present the first genome‐wide association (GWA) and genomic prediction study of Miscanthus that utilizes multilocation phenotypic data. A panel of 568 Miscanthus sinensis accessions was genotyped with 46,177 single nucleotide polymorphisms (SNPs) and evaluated at one subtropical and five temperate locations over 3 years for biomass yield and 14 yield‐component traits. GWA and genomic prediction were performed separately for different years of data in order to assess reproducibility. The analyses were also performed for individual field trial locations, as well as combined phenotypic data across groups of locations. GWA analyses identified 27 significant SNPs for yield, and a total of 504 associations across 298 unique SNPs across all traits, sites, and years. For yield, the greatest number of significant SNPs was identified by combining phenotypic data across all six locations. For some of the other yield‐component traits, greater numbers of significant SNPs were obtained from single site data, although the number of significant SNPs varied greatly from site to site. Candidate genes were identified. Accounting for population structure, genomic prediction accuracies for biomass yield ranged from 0.31 to 0.35 across five northern sites and from 0.13 to 0.18 for the subtropical location, depending on the estimation method. Genomic prediction accuracies of all traits were similar for single‐location and multilocation data, suggesting that genomic selection will be useful for breeding broadly adapted M. sinensis as well as M. sinensis optimized for specific climates. All of our data, including DNA sequences flanking each SNP, are publicly available. By facilitating genomic selection in M. sinensis and Miscanthus × giganteus, our results will accelerate the breeding of these species for biomass in diverse environments.     

‘Collar propagation’ as an alternative propagation method for rhizomatous miscanthus

The demand for perennial nonfood crops, such as miscanthus, is increasing steadily, as fossil resources are replaced by biomass. However, as the establishment of miscanthus is very expensive, its cultivation area in Europe is still small. The most common propagation method for miscanthus is via rhizomes, the harvesting of which is very labour‐intensive. Seed propagation is promising, but not suitable for sterile genotypes. In this study, a new vegetative propagation method, ‘collar propagation’, was tested in field and controlled environment studies. Collars are built at the junction between rhizome and stem. They can be harvested in a less destructive way than rhizomes by pulling out the stems from winter‐dormant miscanthus plants. One genotype of each of the species M. sacchariflorus, M. × giganteus, M. sinensis in combination with three fragment types (collars, rhizomes, collars + rhizomes) were tested for establishment success and plant performance. The performance (e.g. dry matter yield) of collar‐propagated plants was either better than or not significantly different from rhizome‐propagated plants. Pregrown plantlets transplanted into the field showed no significant differences in establishment success between the fragments within a genotype. When directly planted into the field however, the fragment ‘rhizome+collar’ had a significantly better establishment success than the other two. The winter survival rate of the fragment ‘rhizome+collar’ was 70% for M. sacchariflorus and 75% for M. × giganteus. Emergence success from collar‐derived plants was not affected by harvest date (harvested monthly from November to February). This study showed that miscanthus propagation via collars is feasible and a promising alternative to rhizome propagation, as the multiplication rate of collars is comparable to that of rhizome propagation. Collar propagation is the more suitable method for the tested genotypes of the species M. sachariflorus and M. × giganteus, but not for M. sinensis genotypes, which may be better propagated by seeds.     

Winter hardiness of Miscanthus (II): Genetic mapping for overwintering ability and adaptation traits in three interconnected Miscanthus populations

Miscanthus × giganteus (M×g) is the primary species of Miscanthus for bioenergy feedstock production. The current leading biomass cultivar, M×g ‘1993‐1780’, is insufficiently adapted in temperate regions with cold winters such as USDA hardiness zone 5 (average annual minimum temperature of ?28.9 to ?23.3°C) or lower. Three interconnected Miscanthus F₁ populations that shared a common parent were planted in a replicated field trial at Urbana, IL (hardiness zone 5b; average annual minimum temperature of ?26.1 to ?23.3°C) in spring 2011. The winter of 2013–2014 was especially cold in Urbana, with a minimum soil temperature at 10 cm of ?6.2°C and a minimum air temperature of ?25.3°C, giving us an opportunity to evaluate hardiness on established year‐3 plants. The parent in common to all three populations, M. sinensis ssp. condensatus ‘Cosmopolitan’, is native to maritime southern Japan, and in Urbana, it is winter‐damaged most years. In contrast, the three other parents, M. sacchariflorus ‘Robustus’ (MapA), M. sinensis ‘Silberturm’ (MapB), and M. sinensis ‘November Sunset’ (MapC), are typically winter hardy in Urbana. Nearly all MapA progeny plants survived and grew vigorously in spring 2014, whereas in MapB and MapC, many progeny plants did not survive the winter, and most of the survivors were severely damaged, with poor vigor. Negative correlations between overwintering ability and spring regrowth date and autumn dormancy date suggested that the genotypes most likely to survive winters were those that emerged early in spring and/or went dormant early in autumn. Using joint‐population analysis, we identified 53 quantitative trait loci (QTLs) for nine adaptation traits, including nine QTLs for overwintering ability and 11 for spring hardiness scores. Many biologically intuitive candidate genes were observed within or near the QTLs detected in this study, suggesting their validity and potential for further study.     

Invasiveness potential of Miscanthus sinensis: implications for bioenergy production in the United States

Miscanthus sinensis (Anderss.) is a perennial grass species that has been grown widely as an ornamental since the late 1800s and is now being considered for bioenergy production in the United States. With its ability to be grown from seed and tolerate cold climates, this species offers practical advantages over current cultivars of the higher‐yielding hybrid species, M.×giganteus. Yet a large‐scale release of M. sinensis for bioenergy production in colder northern regions could result in new invasions into natural areas. We show, with reference to historical records and data collected in six wild US populations of M. sinensis in 2009, that ornamental varieties of this species have a long history of localized escape in the Eastern United States, primarily within the Appalachian region. To prevent further escape and gene flow, we recommend the development of sterile or functionally sterile varieties of M. sinensis or the restriction of its usage as a donor of genetic material to new sterile cultivars of M. ×giganteus. Other appropriate precautions for new biomass varieties include experimental demonstration of low invasiveness in the target region ahead of commercial production, along with postintroduction stewardship programs.     

Chromosome doubling of the bioenergy crop,Miscanthus×giganteus

The perennial grass, Miscanthus×giganteus is a sterile triploid, which due to its growth rate and biomass accumulation has significant economic potential as a new bioenergy crop. The sterility associated with the triploid genome of this accession requires labor‐intensive vegetative, instead of seed propagation for potential commercial production. Chromosome doubling was used to produce hexaploid plants in an effort to restore fertility to M×giganteus. Tissue culture derived calli from immature inflorescences were treated with the antimitotic agents, colchicine and oryzalin in liquid and solid media. Calli survival rate decreased with increasing concentrations and durations of colchicine or oryzalin treatments and ranged from 0% to 100%. Nuclear DNA content, as determined by flow cytometry, indicated that the frequency of chromosome‐doubled calli varied between compounds and concentrations with the greatest proportion of callus doubling observed using 2‐day treatments of 15 μm oryzalin (78%) or 939 μm colchicine (67%). Liquid media treatments were more effective than solid gels for chromosome doubling. Although oryzalin was effective at chromosome doubling, it inhibited callus growth and plant regeneration frequency. Seven hexaploid plants with doubled DNA content were generated, which displayed increased stomata size (30.0±0.2 μm) compared with regenerated triploid M. ×giganteus plants (24.3±1.0 μm). Following clonal replication these plants will be evaluated for growth rate, biomass accumulation, and pollen viability. Successful chromosome doubling and plant regeneration of M.×giganteus suggests that ploidy manipulation of this plant and its parental species (Miscanthus sinensis and Miscanthus sacchariflorus) could be a means to access genetic variability for the improvement of Miscanthus as a biofuel/bioenergy crop.     

Effects of temperature,illumination and node position on stem propagation of Miscanthus × giganteus

The sterile triploid Miscanthus × giganteus is capable of yielding more biomass per unit land area than most other temperate crops. Although the yield potential of M. × giganteus is high, sterility requires all propagation of the plant to be done vegetatively. The traditional rhizome propagation system achieves relatively low multiplication rates, i.e. the number of new plants generated from a single‐parent plant, and requires tillage that leaves soil vulnerable to CO₂ and erosion losses. A stem‐based propagation system is used in related crops like sugarcane, and may prove a viable alternative, but the environmental conditions required for shoot initiation from stems of M. × giganteus are unknown. A study was conducted to investigate the effect of temperature, illumination and node position on emergence of M. × giganteus shoots. Stems of M. × giganteus were cut into segments with a single node each, placed in controlled environments under varied soil temperature or light regimes and the number of emerged shoots were evaluated daily for 21 days. At temperatures of 20 and 25 °C, rhizomes produced significantly more shoots than did stem segments (P = 0.0105 and 0.0594, respectively), but the difference was not significant at 30 °C, where 63% of stems produced shoots compared to 80% of rhizomes (P = 0.2037). There was a strong positive effect (P = 0.0086) of soil temperature on emergence in the range of temperatures studied here (15–30 °C). Node positions higher on the stem were less likely to emerge (P < 0.0001) with a significant interaction between illumination and node position. Planting the lowest five nodes from stems of M. × giganteus in 30 °C soil in the light resulted in 75% emergence, which represents a potential multiplication rate 10–12 times greater than that of the current rhizome‐based system.     

Accumulation of soil organic carbon after cropland conversion to short‐rotation willow and poplar

The demand for bioenergy has increased the interest in short‐rotation woody crops (SRWCs) in temperate zones. With increased litter input and ceased annual soil cultivation, SRWC plantations may become soil carbon sinks for climate change mitigation. A chronosequence of 26 paired plots was used to study the potential for increasing soil organic carbon (SOC) under SRWC willow and poplar after conversion from cropland (CR) on well‐drained soils. We estimated SOC stocks in SRWC stands and adjacent CR and related the difference to time since conversion, energy crop species, SOC stock of the adjacent CR (proxy for initial SOC of SRWC) and the fine soil percentage (<63 μm) (FS). Soil cores to 40 cm depth were sampled and separated by layers of fixed depths (0–5, 5–10, 10–15, 15–25 and 25–40 cm). Additionally, soils were sampled from soil pits by genetic horizons to 100 cm depth. Comparisons of SOC stocks by equivalent soil masses showed that mean SOC stocks in SRWC were 1.7 times higher than those of CR in the top 5 cm of the soil (P < 0.001). The differences between SRWC and CR remained significant for the plough layer (0–25 cm) by a factor of 1.2 (P = 0.003), while no changes were detectable for the 0–40 cm (P = 0.32), or for the entire 0–100 cm soil layer (P = 0.29). The SOC stock ratio, that is the ratio of SOC stock in SRWC relative to CR, did not change significantly with time since conversion, although there was a tendency to an increase over time for the top 40 cm (P = 0.09). The SOC stock ratio was negatively correlated to SOC in CR and FS percentage, but there was no significant difference between willow and poplar at any depth. Our results suggest that SOC stocks in the plough layer increase after conversion to SRWC.     

Radiation capture and conversion efficiencies of Miscanthus sacchariflorus,M. sinensis and their naturally occurring hybrid M. × giganteus

Miscanthus is a rhizomatous C4 grass of great interest as a biofuel crop because it has the potential to produce high yields over a wide geographical area with low agricultural inputs on marginal land less suitable for food production. At the moment, a clonal interspecific hybrid Miscanthus × giganteus is the most widely cultivated and studied in Europe and the United States, but breeding programmes are developing newer more productive varieties. Here, we quantified the physiological processes relating to whole season yield in a replicated plot trial in Wales, UK. Light capture and conversion efficiency were parameterized for four carefully selected genotypes (M. sinensis, M. sacchariflorus and Miscanthus × giganteus). Differences in the canopy architecture in mature stands as measured by the extinction coefficient (k) were small (0.55–0.65). Sensitivity analysis on a mathematical model of Miscanthus was performed to quantify the accumulative intercepted photosynthetically active radiation (iPAR) in the growing season using (i) k, (ii) variation in the thermal responses of leaf expansion rate, (iii) base temperature for degree days and (iv) date start of canopy expansion. A 10% increase in k or leaf area per degree day both had a minimal effect on iPAR (3%). Decreasing base temperature from 10 to 9 °C gave an 8% increase in iPAR. If the starting date for canopy expansion was the same as shoot emergence date, then the iPAR increases by 12.5%. In M. × giganteus, the whole season above ground and total (including below ground) radiation‐use efficiency (RUE) ranged from 45% to 37% higher than the noninterspecific hybrid genotypes. The greater yields in the interspecific hybrid M. × giganteus are explained by the higher RUE and not by differences in iPAR or partitioning effects. Studying the mechanisms underlying this complex trait could have wide benefits for both fuel and food production.     

Long‐term yield and performance of 15 different Miscanthus genotypes in southwest Germany

A field experiment with 15 Miscanthus genotypes including M. × giganteus, M. sacchariflorus, M. sinensis and M. sinensis hybrids was conducted for 14 years at the experimental Ihinger Hof station of the University of Hohenheim in southwest Germany to evaluate interannual yield performance stability over 14 years of harvests of the different genotypes. In this article, a simple formula is presented which could be used to forecast late winter yields using morphological traits in autumn. The data obtained indicated a shorter establishment period to reach a yield plateau of M. × giganteus and M. sacchariflorus than M. sinensis hybrids and M. sinensis genotypes. The best performing genotype was M. × giganteus (Gig‐2, No. 16.21) with a mean spring harvestable yield of 14.1 t DM ha^?1 year^?1. A correlation analysis with climatic parameters revealed precipitation during the growing period as the key factor for high yields at this site. Likewise, but to a lesser degree, heat sum during the growing period was positively correlated with yields. It could be shown that precipitation/snow during the winter correlated with yield losses, until the harvest date in February/March. Phenological measurements indicated that a high yield potential of the tested genotypes is associated with either an absence of flowering or late flowering. Also, height of the plants and shoot diameter were indicators for high yield potential. Shoot density and plant height at senescence were found to be solid parameters to estimate harvestable biomass in late winter. Yield approximations with a mean accuracy of 80.9% for M.× giganteus genotypes were obtained using the newly developed equation. Yields of M. sinensis hybrid (Sin‐H7) were projected most accurately with this simple formula, resulting in a mean accuracy of 84.5%.