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.     

Plant morphology,genome size,and SSR markers differentiate five distinct taxonomic groups among accessions in the genus Miscanthus

Information on genome size, ploidy level, and genomic polymorphisms among accessions of the genus Miscanthus can assist in taxonomic studies, help understand the evolution of the genus, and provide valuable information to biomass crop improvement programs. Taxonomic investigation combining variation in plant morphology, genome size, chromosome numbers, and simple sequence repeat (SSR) marker polymorphisms were applied to characterize 101 Miscanthus accessions. A total of 258 amplicons generated from 17 informative SSR primer pairs was subjected to cluster and principal coordinate analysis and used to characterize genetic variation and relationships among 31 Miscanthus accessions, including four interspecific Miscanthus hybrids created from controlled pollinations, and four Saccharum, six Erianthus, and one Sorghum bicolor accessions. Miscanthus accessions were distinct from accessions in the genera Erianthus and Saccharum. Miscanthus accessions fell into five taxonomic groups, including the existing taxonomic section Miscanthus, diploid and tetraploid Miscanthus sacchariflorus, and a fourth (M. × giganteus) and fifth group (Miscanthus ‘purpurascens’); the last two being intermediate forms. In contrast to previous work, our findings suggest diploid and tetraploid M. sacchariflorus are taxonomically different, the latter more closely related to M. sacchariflorus var lutarioriparius. We also suggest that Miscanthus ‘purpurascens’ accessions are interspecific hybrids between Miscanthus sinensis and diploid M. sacchariflorus based on DNA content and SSR polymorphisms. The evolution of Miscanthus and related genera is discussed based on combined analysis and geographical origin.     

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.     

Genetic Analysis of Putative Triploid Miscanthus Hybrids and Tetraploid M. sacchariflorus Collected from Sympatric Populations of Kushima, Japan

Miscanthus ?×giganteus, a triploid hybrid between tetraploid M. sacchariflorus and diploid M. sinensis, has considerable potential as a bioenergy crop. Currently only one genotype is widely cultivated, increasing its vulnerability to diseases during production. Finding new hybrids is important to broaden genetic resources of M. ×giganteus. Three putative triploid hybrids were discovered in a sympatric population of tetraploid M. sacchariflorus and diploid M. sinensis in Kushima, Japan. The hybrid nature of the triploids was determined by morphological analysis and sequencing the ribosomal DNA internal transcribed spacer (ITS) region. The triploids had awns on their florets, which is a common characteristic of diploid M. sinensis, and sheath hairs, which is typical of tetraploid M. sacchariflorus. All triploids showed heterozygosity in their ribosomal DNA ITS sequences. Based on these results, it is confirmed that the triploids are hybrids and novel genotypes of M. ×giganteus. Natural crossing between tetraploid M. sacchariflorus × diploid M. sinensis may also lead to the production of tetraploid hybrids. ITS analysis of tetraploid plants showed that one maternal parent of the triploid hybrids, K-Ogi-1, had heterozygous ITS, which was different than the other analyzed tetraploid, M. sacchariflorus. Thus, K-Ogi-1 was likely of hybrid origin. These tetraploid hybrids can also be utilized as parents in M. ×giganteus breeding. Since all hybrids identified in this study had tetraploid M. sacchariflorus as maternal parents, collecting and analyzing seeds from tetraploid M. sacchariflorus in sympatric areas could be an effective strategy to identify natural Miscanthus hybrids that can be used as bioenergy crops.     

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.     

Genetic Diversity and Population Structure of Miscanthus sinensis Germplasm in China

Miscanthus is a perennial rhizomatous C4 grass native to East Asia. Endowed with great biomass yield, high ligno-cellulose composition, efficient use of radiation, nutrient and water, as well as tolerance to stress, Miscanthus has great potential as an excellent bioenergy crop. Despite of the high potential for biomass production of the allotriploid hybrid M. ×giganteus, derived from M. sacchariflorus and M. sinensis, other options need to be explored to improve the narrow genetic base of M. ×giganteus, and also to exploit other Miscanthus species, including M. sinensis (2n = 2x = 38), as bioenergy crops. In the present study, a large number of 459 M. sinensis accessions, collected from the wide geographical distribution regions in China, were genotyped using 23 SSR markers transferable from Brachypodium distachyon. Genetic diversity and population structure were assessed. High genetic diversity and differentiation of the germplasm were observed, with 115 alleles in total, a polymorphic rate of 0.77, Nei’s genetic diversity index (He) of 0.32 and polymorphism information content (PIC) of 0.26. Clustering of germplasm accessions was primarily in agreement with the natural geographic distribution. AMOVA and genetic distance analyses confirmed the genetic differentiation in the M. sinensis germplasm and it was grouped into five clusters or subpopulations. Significant genetic variation among subpopulations indicated obvious genetic differentiation in the collections, but within-subpopulation variation (83%) was substantially greater than the between-subpopulation variation (17%). Considerable phenotypic variation was observed for multiple traits among 300 M. sinensis accessions. Nine SSR markers were found to be associated with heading date and biomass yield. The diverse Chinese M. sinensis germplasm and newly identified SSR markers were proved to be valuable for breeding Miscanthus varieties with desired bioenergy traits.     

Nuclear DNA content variation of three Miscanthus species in China

In order to estimate the variation in nuclear genome size in Miscanthus, flow cytometry of nuclei stained by propidium iodide was carried out using 36 populations of three Miscanthus species: M. lutarioriparius, M. sacchariflorus and M. sinensis, which were sampled from cold northern to warm and humid southern and central China, as well as near the sea level in eastern China to mountains in western China. The DNA content of diploid was 4.37 ± 0.02 pg/2C in M. lutarioriparius, 4.37 ± 0.01 pg/2C in M. sacchariflorus, and 5.37 ± 0.03 pg/2C in M. sinensis, respectively. There was no intraspecific variation in the three Miscanthus species at the diploid level, suggesting that the genome size was stable within species and the diverse environments did not induce variation in genome size at the diploid level. However, tetraploid populations were found in M. lutarioriparius and M. sacchariflorus, and their genome sizes were 8.56 and 8.54 pg, respectively, which are lower than expected values (8.74 pg), indicating the genome downsizing after polyploidization in the genus. Our results showed that the plant height of M. lutarioriparius was the highest one among the three species and the species was more closely related to M. sacchariflorus than M. sinensis. The intra-species genomic variation and inter-species differentiation in Miscanthus species provide important genetic and genomic information for the development of Miscanthus, especially for the endemic species, M. lutarioriparius, (together with Miscanthus × giganteus) which are now emerging as a key bio-energy crop because of their high yields and strong adaptability.     

SSR-based genetic maps of Miscanthus sinensis and M. sacchariflorus, and their comparison to sorghum

We present SSR-based genetic maps from a cross between Miscanthus sacchariflorus Robustus and M. sinensis, the progenitors of the promising cellulosic biofuel feedstock Miscanthus × giganteus. cDNA-derived SSR markers were mapped by the two-way pseudo-testcross model due to the high heterozygosity of each parental species. A total of 261 loci were mapped in M. sacchariflorus, spanning 40 linkage groups and 1,998.8 cM, covering an estimated 72.7% of the genome. For M. sinensis, a total of 303 loci were mapped, forming 23 linkage groups and 2,238.3 cM, covering 84.9% of the genome. The use of cDNA-derived SSR loci permitted alignment of the Miscanthus linkage groups to the sorghum chromosomes, revealing a whole genome duplication affecting the Miscanthus lineage after the divergence of subtribes Sorghinae and Saccharinae, as well as traces of the pan-cereal whole genome duplication. While the present maps provide for many early research needs in this emerging crop, additional markers are also needed to improve map density and to further characterize the structural changes of the Miscanthus genome since its divergence from sorghum and Saccharum.     

Biomass yield in a genetically diverse Miscanthus sacchariflorus germplasm panel phenotyped at five locations in Asia,North America,and Europe

Miscanthus is a high-yielding bioenergy crop that is broadly adapted to temperate and tropical environments. Commercial cultivation of Miscanthus is predominantly limited to a single sterile triploid clone of Miscanthus × giganteus, a hybrid between Miscanthus sacchariflorus and M. sinensis. To expand the genetic base of M. × giganteus, the substantial diversity within its progenitor species should be used for cultivar improvement and diversification. Here, we phenotyped a diversity panel of 605 M. sacchariflorus from six previously described genetic groups and 27 M. × giganteus genotypes for dry biomass yield and 16 yield-component traits, in field trials grown over 3 years at one subtropical location (Zhuji, China) and four temperate locations (Foulum, Denmark; Sapporo, Japan; Urbana, Illinois; and Chuncheon, South Korea). There was considerable diversity in yield and yield-component traits among and within genetic groups of M. sacchariflorus, and across the five locations. Biomass yield of M. sacchariflorus ranged from 0.003 to 34.0 Mg ha⁻¹ in year 3. Variation among the genetic groups was typically greater than within, so selection of genetic group should be an important first step for breeding with M. sacchariflorus. The Yangtze 2x genetic group (=ssp. lutarioriparius) of M. sacchariflorus had the tallest and thickest culms at all locations tested. Notably, the Yangtze 2x genetic group's exceptional culm length and yield potential were driven primarily by a large number of nodes (>29 nodes culm⁻¹ average over all locations), which was consistent with the especially late flowering of this group. The S Japan 4x, the N China/Korea/Russia 4x, and the N China 2x genetic groups were also promising genetic resources for biomass yield, culm length, and culm thickness, especially for temperate environments. Culm length was the best indicator of yield potential in M. sacchariflorus. These results will inform breeders' selection of M. sacchariflorus genotypes for population improvement and adaptation to target production environments.     

Characterization of flowering time diversity in Miscanthus species

Miscanthus sinensis, M. sacchariflorus and their hybrids have been identified as leading candidates for the provision of bioenergy production across several continents. Flowering time is an important trait affecting biomass yield as well as certain quality attributes, such as moisture content at harvest. The aim of this study was to ascertain the level of diversity available to breeders and potential for hybridisation of different accessions in a large collection of Miscanthus. We also sought to determine trends in flowering time within and between species with respect to environment and origin of collection data (where known), whether flowering order was maintained across years, and the extent of uniformity of flowering in different genotypes. Flowering time was observed weekly in 244 genotypes of two Miscanthus species (M. sinensis, M. sacchariflorus) and inter‐specific hybrids including M. x giganteus over 3 years and using 4 clonal replicates of each genotype on a trial planted near Aberystwyth (Wales, UK). Differences in flowering time across the entire collection ranged from 160 to 334 days (June to November) and photoperiods between 7.8 and 16.6 h, with associated accumulated temperatures of 161 to 865^oCd. More than two thirds of the collection flowered by the end of each growing season. M. sinensis individuals were the earliest genotypes to flower and showed the greatest diversity with respect to the onset of flowering. Flowering times in genotypes of known origin in Asia could be partially explained by growing season rain fall, degree days and mean temperature. Uniform flowering was identified in some genotypes. This will be important for the development of genetically diverse seed‐based crops. Rank order of flowering was shown to be consistent across Western Europe, thereby justifying single site trials as the basis of germplasm characterisation for wider geographical deployment.     

Shoot organogenesis in three Miscanthus species and evaluation for genetic uniformity using AFLP analysis

A simple, efficient protocol for direct in vitro shoot organogenesis and regeneration was established for three species of Miscanthus including two clones of Miscanthus x giganteus, one clone of M. sinensis and one clone of M. sacchariflorus. Shoots were induced from the axillary nodes of both M. x giganteus and M. sacchariflorus and from apical meristems of both M. sinensis and M. sacchariflorus. A tillering method was used to accelerate shoot proliferation. Shoots were rooted in a wet perlite substrate in pots in the greenhouse. Subsequently, rooted plants were transferred to the field. The genetic uniformity of regenerated plants was evaluated using amplified fragment length polymorphism analysis and compared to that of rhizome-propagated plants. A total of 33,443 fragments were generated, representing 869 markers. There were 21 fragments (0.06 % of the fragments) or 19 markers (2.19 % of the markers) that were polymorphic, and almost all of these were singletons. The three species showed similar polymorphisms. Genetic variability was also found in the rhizome-propagated plants, sometimes at a higher rate than in the in vitro culture, indicating that the genetic uniformity was not altered by the protocol. This protocol may help breeders produce new clones of Miscanthus in the future.     

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.     

Water Use Efficiency and Biomass Partitioning of Three Different Miscanthus Genotypes with Limited and Unlimited Water Supply

Miscanthus species, which are C₄perennial grasses, have a highbiomass potential but yields at many sites in Europe can belimited by insufficient water supply and plant survival is endangeredunder extreme summer drought. A pot experiment was conductedto measure the influence of reduced water supply on the wateruse efficiency (WUE) and biomass partitioning of three Miscanthusgenotypes (M. x giganteus, M. sacchariflorus, and a M. sinensishybrid) in a controlled environment. The experiment consistedof three phases (phase 1 = 0–20 d; phase 2 = 21–39d; phase 3 = 40–54 d) punctuated by destructive harvests.In phase 1, soil moisture was non-limiting. In the second andthird phases, lowered soil moisture contents induced water deficits.Air vapour pressure deficit (VPD) was 0.49 ± 0.05 kPa.Water deficits caused leaf senescence in M. x giganteus andM. sacchariflorus, but not in the M. sinensis hybrid. Greenleaf conductances were lowest in M. sinensis under water deficit,indicating stomatal regulation. Water use efficiency for wholeplants of each genotype ranged from 11.5 to 14.2 g dry matter(DM) kg^-1H₂O but did not differ significantly between genotypesor water treatments under the conditions of this experiment.However, differences in dry matter partitioning to the shoot(the harvestable component) resulted in genotypic differencesin WUE, calculated on a harvestable dry matter basis, whichranged from 4.1 g DM kg^-1H₂O for M. sacchariflorus to 2.2 gDM kg^-1H₂O for M. x giganteus. Copyright 2000 Annals of BotanyCompany Miscanthus sinensis, Miscanthus sacchariflorus, Miscanthus x giganteus, water use efficiency, biomass, C₄plants, drought     

Phenotypic Variation in Senescence in Miscanthus: Towards Optimising Biomass Quality and Quantity

Senescence impacts the harvestable biomass yield and quality in Miscanthus. Very early autumn senescence shortens the canopy duration reducing yield potential. When senescence is too late or slow, the crop does not ripen sufficiently before harvest, resulting in high moisture and nutrient offtakes that reduce biomass quality. In this study, variation in senescence was monitored over 3?years in a trial of 244 Miscanthus genotypes planted in four replicate blocks. The experiment comprised 199 genotypes of Miscanthus sinensis, 36 genotypes of Miscanthus sacchariflorus, and 9 genotypes of Miscanthus?×?giganteus. On average, M. sinensis genotypes remained greener for longer than M. sacchariflorus genotypes. There was a strong correlation between senescence and moisture content at harvest in 2007 and 2008 (R ²?=?0.59, R ²?=?0.57, respectively; n?=?244). The senescence rate of M.?×?giganteus, an interspecific hybrid between M. sinensis and M. sacchariflorus, was found to lie between the two parental species groups. Environmental signals likely to be involved in the timing and rate of senescence, such as variation in photoperiod and thermal time that occurred through the growing season were investigated. Interactions between individual environmental signals and senescence were difficult to separate. The majority of plants senesced consistently between replicate blocks, and rank order was mostly consistent across all years suggesting strong genotypic control of senescence. This study provides valuable information for the future optimisation of Miscanthus, and potentially other energy grasses, where new varieties are needed to maximise net energy yields and crop quality for different end uses in different global regions.     

Chilling tolerance and early vigour-related characteristics evaluated in two Miscanthus genotypes

A long growing season, mediated by the ability to grow at low temperatures early in the season, can result in higher yields in biomass of crop Miscanthus. In this paper, the chilling tolerance of two highly productive Miscanthus genotypes, the widely planted Miscanthus × giganteus and the Miscanthus sinensis genotype ‘Goliath’, was studied. Measurements in the field as well as under controlled conditions were combined with the main purpose to create basic comparison tools in order to investigate chilling tolerance in Miscanthus in relation to its field performance. Under field conditions, M. × giganteus was higher yielding and had a faster growth rate early in the growing season. Correspondingly, M. × giganteus displayed a less drastic reduction of the leaf elongation rate and of net photosynthesis under continuous chilling stress conditions in the growth chamber. This was accompanied by higher photochemical quenching and lower nonphotochemical quenching in M. × giganteus than that in M. sinensis ‘Goliath’ when exposed to chilling temperatures. No evidence of impaired stomatal conductance or increased use of alternative electron sinks was observed under chilling stress. Soluble sugar content markedly increased in both genotypes when grown at 12°C compared to 20°C. The concentration of raffinose showed the largest relative increase at 12°C, possibly serving as a protection against chilling stress. Overall, both genotypes showed high chilling tolerance for C₄ plants, but M. × giganteus performed better than M. sinensis ‘Goliath’. This was not due to its capacity to resume growth earlier in the season but rather due to a higher growth rate and higher photosynthetic efficiency at low temperatures.     

Miscanthus sacchariflorus – biofuel parent or new weed?

The perennial grass triploid Miscanthus × giganteus is a promising renewable bioenergy feedstock in the United States and Europe. Originating from eastern Asia, this species is a sterile hybrid cross between M. sinensis and M. sacchariflorus. While research has begun to examine the impacts of M. sinensis and triploid M. × giganteus on the landscape, M. sacchariflorus has been largely overlooked in the peer‐reviewed literature. This review article discusses the origin, uses, distribution, and invasive potential of M. sacchariflorus. M. sacchariflorus is capable of producing high yields (10.7 t DM ha^?1 yr^?1), generally does not reproduce by seed, and can be challenging to establish due to poor cold tolerance, likely due to the limited germplasm used in evaluations. However, M. sacchariflorus has abundant and aggressively spreading rhizomes, which underscores its invasive risk. In the United States, it is listed as escaped from cultivation in at least eight states, primarily in the Midwest, although it is likely that not all populations have been reported. As such, it is essential to generate a comprehensive dataset of all known M. sacchariflorus populations and monitor any continued spread of this species.     

Variability and adaptability of Miscanthus species evaluated for energy crop domestication

A growing body of evidence indicates that second‐generation energy crops can play an important role in the development of renewable energy and the mitigation of climate change. However, dedicated energy crops have yet to be domesticated in order to fully realize their productive potential under unfavorable soil and climatic conditions. To explore the possibility of domesticating Miscanthus crops in northern China where marginal and degraded land is abundant, we conducted common garden experiments at multiple locations to evaluate variation and adaptation of three Miscanthus species that are likely to serve as the wild progenitors of the energy crops. A total of 93 populations of Miscanthus sinensis, Miscanthus sacchariflorus, and Miscanthus lutarioriparius were collected across their natural distributional ranges in China and grown in three locations that represent temperate grassland with cold winter, the semiarid Loess Plateau, and relatively warm and wet central China. Evaluated with growth traits such as plant height, tiller number, tiller diameter, and flowering time, the Miscanthus species showed high levels of genetic variation within and between species. There were significant site × population interactions for almost all traits of M. sacchariflorus and M. sinensis, but not M. lutarioriparius. The northern populations of M. sacchariflorus had the highest establishment rates at the most northern site owing to their strong cold tolerance. An endemic species in central China, M. lutarioriparius, produced not only the highest biomass of the three species but also higher biomass at the Loess Plateau than the southern site near its native habitats. These results demonstrated that the wild species harbored a high level of genetic variation underlying traits important for crop establishment and production at sites that are colder and drier than their native habitats. The natural variation and adaptive plasticity found in the Miscanthus species indicated that they could provide valuable resources for the development of second‐generation energy crops.     

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%.     

Microsatellites and RFLP probes from maize are efficient sources of molecular markers for the biomass energy crop Miscanthus

A survey of Gramineae markers was carried out with the aim of developing cost-effective methods for the molecular analysis of Miscanthus species. Ten out of twenty Gramineae RFLP probes from ”anchor” sets hybridized well to Miscanthus DNA while all 15 maize probes tested cross-hybridized successfully, showing similar patterns in both species. Cross-taxa amplification of maize microsatellite primers was then tested. This showed that 57 out of 76 (75%) give highly reproducible amplification with Miscanthus DNA. Amplification products differed in size from those in maize but there was no bias toward higher or lower molecular weights. Microsatellite polymorphism produced by 17 primer pairs was studied in detail in a panel of 11 Miscanthus clones belonging to the species Miscanthus sinensis, Miscanthus sacchariflorus, Miscanthus ×giganteus and Miscanthus condensatus. Intra- and inter-specific length polymorphisms were frequent between the tested Miscanthus clones with length polymorphisms being found for all primer pairs, detecting 3–22 alleles. Polymorphism information content (PIC) values for microsatellites ranged from 0.48 to 0.94 with an average of 0.83. Species-specific amplicons were produced by two microsatellites. Genetic similarity coefficients of the Miscanthus clones ranged from 0.35 to 0.92, with an average of 0.57. Five polymorphisms were studied in a segregating population, where they showed Mendelian inheritance. In addition, two microsatellite markers mapping 1.3-cM apart on maize chromosome 7 were linked in Miscanthus at an estimated distance of 8 cM, suggesting collinearity. The high transferability of microsatellite markers from maize will enhance the power and resolution of genome analysis in Miscanthus. Received: 14 April 2000 / Accepted: 9 June 2000