Transferability of Microsatellite Markers from Brachypodium distachyon to Miscanthus sinensis,a Potential Biomass Crop

Miscanthus sinensis has high biomass yield and contributed two of the three genomes in M.x giganteus,a bioenergy crop widely studied in Europe and North America,and thus is a potential biomass crop and an important germplasm for Miscanthus breeding.Molecular markers are essential for germplasm evaluation,genetic analyses and new cultivar development in M.sinensis.In the present study,we reported transferability of simple sequence repeat (SSR) markers from Brachypodium distachyon to M.sinensis.A set of 57 SS...     

A semimechanistic model predicting the growth and production of the bioenergy crop Miscanthus×giganteus: description, parameterization and validation

Biomass based bioenergy is promoted as a major sustainable energy source which can simultaneously decrease net greenhouse gas emissions. Miscanthus × giganteus ( M. × giganteus ), a C₄ perennial grass with high nitrogen, water, and light use efficiencies, is regarded as a promising energy crop for biomass production. Mathematical models which can accurately predict M. × giganteus biomass production potential under different conditions are critical to evaluate the feasibility of its production in different environments. Although previous models based on light-conversion efficiency have been shown to provide good predictions of yield, they cannot easily be used in assessing the value of physiological trait improvement or ecosystem processes. Here, we described in detail the physical and physiological processes of a previously published generic mechanistic eco-physiological model, WIMOVAC, adapted and parameterized for M. × giganteus . Parameterized for one location in England, the model was able to realistically predict daily field diurnal photosynthesis and seasonal biomass at a range of other sites from European studies. The model provides a framework that will allow incorporation of further mechanistic information as it is developed for this new crop.     

Transferability of microsatellite markers from Brachypodium distachyon to Miscanthus sinensis, a potential biomass crop

Miscanthus sinensis has high biomass yield and contributed two of the three genomes in M. x giganteus, a bioenergy crop widely studied in Europe and North America, and thus is a potential biomass crop and an important germplasm for Miscanthus breeding. Molecular markers are essential for germplasm evaluation, genetic analyses and new cultivar development in M. sinensis. In the present study, we reported transferability of simple sequence repeat (SSR) markers from Brachypodium distachyon to M. sinensis. A set of 57 SSR markers evenly distributed across the B. distachyon genome were deliberately designed. Out of these B. distachyon SSR markers, 86.0% are transferable to M. sinensis. The SSR loci amplified in M. sinensis were validated by re-sequencing the amplicons. The polymorphism information content (PIC) of the transferable SSR markers varied from 0.073 to 0.375 with a mean of 0.263, assessed based on 21 M. sinensis genotypes. Phylogenetic tree based on 162 alleles detected by 49 SSR markers could unambiguously distinguish B. distachyon from M. sinensis, and cluster 21 M. sinensis genotypes into three groups that are basically in coincidence with their geographical distribution and ecotype classifications. The markers developed by the comparative genomic approach could be useful for germplasm evaluation, genetic analysis, and marker-assisted breeding in Miscanthus.     

Overwintering problems of newly established Miscanthus plantations can be overcome by identifying genotypes with improved rhizome cold tolerance

Miscanthus , a perennial rhizomatous C₄ grass, is a potential biomass crop in Europe, mainly because of its high yield potential and low demand for inputs. However, until recently only a single clone, M. × giganteus , was available for the extensive field trials performed across Europe and this showed poor overwintering in the first year after planting at some locations in Northern Europe. Therefore, field trials with five Miscanthus genotypes, including two acquisitions of Miscanthus × giganteus , one of M. sacchariflorus and two hybrids of M. sinensis were planted in early summer 1997 at four sites, in Sweden, Denmark, England and Germany. The field trials showed that better overwintering of newly established plants at a site was not apparently connected with size or early senescence. An artificial freezing test with rhizomes removed from the field in January 1998 showed that the lethal temperature at which 50% were killed (LT₅₀) for M. × giganteus and M. sacchariflorus genotypes was −3.4 °C. However, LT₅₀ in one of the M. sinensis hybrid genotypes tested was −6.5 °C and this genotype had the highest survival rates in the field in Sweden and Denmark. Although the carbohydrate content of rhizomes, osmotic potential of cell sap and mineral composition were not found to explain differences in frost tolerance adequately, moisture contents correlated with frost hardiness (LT₅₀) in most cases. The results obtained form a basis for identifying suitable Miscanthus genotypes for biomass production in the differing climatic regions of Europe.     

Development of SCAR marker for simultaneous identification of Miscanthus sacchariflorus, M. sinensis and M. x giganteus

The sequence-characterized amplified region (SCAR) marker for simultaneous identification of Miscanthus sacchariflorus, Miscanthus sinensis, and Miscanthus x giganteus was developed. In this study, it was attempted for the first time to develop the SCAR marker for detecting the molecular phenotypes among Miscanthus species. Randomly amplified polymorphic DNA technique was applied for this study and one fragment which is unique to M. sacchariflorus was identified and then sequenced. Based on the specific fragment, one SCAR primer pair designated as MS62-5F and MS62-5R was designed to amplify an approximately 1,000 bp DNA fragment within the sequenced region. Diagnostic PCR was performed using the primer pair. Using this SCAR marker, approximately 1,000 bp and 1,200 bp DNA fragments were obtained in M. sacchariflorus and M. sinensis, respectively. Moreover, M. x giganteus was obtained both bands at the same time. The result showed that this SCAR marker can clearly distinguish the M. sacchariflorus, M. sinensis, and M. x giganteus, respectively.     

Greenhouse gas emissions from four bioenergy crops in England and Wales: Integrating spatial estimates of yield and soil carbon balance in life cycle analyses

Accurate estimation of the greenhouse gas (GHG) mitigation potential of bioenergy crops requires the integration of a significant component of spatially varying information. In particular, crop yield and soil carbon (C) stocks are variables which are generally soil type and climate dependent. Since gaseous emissions from soil C depend on current C stocks, which in turn are related to previous land management it is important to consider both previous and proposed future land use in any C accounting assessment. We have conducted a spatially explicit study for England and Wales, coupling empirical yield maps with the RothC soil C turnover model to simulate soil C dynamics. We estimate soil C changes under proposed planting of four bioenergy crops, Miscanthus ( Miscanthus × giganteus ), short rotation coppice (SRC) poplar ( Populus trichocarpa Torr. & Gray × P. trichocarpa , var. Trichobel), winter wheat, and oilseed rape. This is then related to the former land use – arable, pasture, or forest/seminatural, and the outputs are then assessed in the context of a life cycle analysis (LCA) for each crop. By offsetting emissions from management under the previous land use, and considering fossil fuel C displaced, the GHG balance is estimated for each of the 12 land use change transitions associated with replacing arable, grassland, or forest/seminatural land, with each of the four bioenergy crops. Miscanthus and SRC are likely to have a mostly beneficial impact in reducing GHG emissions, while oilseed rape and winter wheat have either a net GHG cost, or only a marginal benefit. Previous land use is important and can make the difference between the bioenergy crop being beneficial or worse than the existing land use in terms of GHG balance.     

The use of dna sequencing (ITS and trnL-F), AFLP, and fluorescent in situ hybridization to study allopolyploid Miscanthus (Poaceae)

Two clones of Miscanthus, grown under the names M. ×giganteus and M. sacchariflorus, have been used in biomass trials in Europe, but neither the identity of these clones nor their origin has been established. DNA sequencing, amplified fragment length polymorphism (AFLP), and chromosome studies confirm that M. ×giganteus is an allotriploid (2n = 3x = 57) combining genomes from M. sinensis (2n = 2x = 38) and M. sacchariflorus (2n = 38 or 76). Two alleles of the internal transcribed spacer of 18S-25S nuclear ribosomal DNA (ITS) were discovered in polymerase chain reaction products of M. ×giganteus. Cloning of these revealed that one matched M. sinensis and the other M. sacchariflorus. Plastid trnL intron and trnL-F spacer sequences showed that the maternal lineage of M. ×giganteus was M. sacchariflorus. Fluorescent in situ hybridization, FISH, was used to investigate genome organization in Miscanthus but was unable to differentiate between the different parental genomes present in M. ×giganteus, indicating that two parental genomes are still extremely similar at the repetitive DNA level. This study is an example in which rDNA sequences and AFLP fingerprints permit identification of the parental genomes in a hybrid, but FISH methods, at the repetitive DNA level (including genomic in situ hybridization, GISH), were unable to do so because their sequences remain too similar.     

High resolution genetic mapping by genome sequencing reveals genome duplication and tetraploid genetic structure of the diploid Miscanthus sinensis

We have created a high-resolution linkage map of Miscanthus sinensis, using genotyping-by-sequencing (GBS), identifying all 19 linkage groups for the first time. The result is technically significant since Miscanthus has a very large and highly heterozygous genome, but has no or limited genomics information to date. The composite linkage map containing markers from both parental linkage maps is composed of 3,745 SNP markers spanning 2,396 cM on 19 linkage groups with a 0.64 cM average resolution. Comparative genomics analyses of the M. sinensis composite linkage map to the genomes of sorghum, maize, rice, and Brachypodium distachyon indicate that sorghum has the closest syntenic relationship to Miscanthus compared to other species. The comparative results revealed that each pair of the 19 M. sinensis linkages aligned to one sorghum chromosome, except for LG8, which mapped to two sorghum chromosomes (4 and 7), presumably due to a chromosome fusion event after genome duplication. The data also revealed several other chromosome rearrangements relative to sorghum, including two telomere-centromere inversions of the sorghum syntenic chromosome 7 in LG8 of M. sinensis and two paracentric inversions of sorghum syntenic chromosome 4 in LG7 and LG8 of M. sinensis. The results clearly demonstrate, for the first time, that the diploid M. sinensis is tetraploid origin consisting of two sub-genomes. This complete and high resolution composite linkage map will not only serve as a useful resource for novel QTL discoveries, but also enable informed deployment of the wealth of existing genomics resources of other species to the improvement of Miscanthus as a high biomass energy crop. In addition, it has utility as a reference for genome sequence assembly for the forthcoming whole genome sequencing of the Miscanthus genus.     

Miscanthus: A fast‐growing crop for environmental remediation and biofuel production

As an herbaceous perennial, Miscanthus has attracted extensive attention in bioenergy refinery and ecological remediation due to its high yield and superior environmental adaptability. This review summarizes current research advances of Miscanthus in several aspects including biological properties, biofuels production, and phytoremediation of contaminated soil. Miscanthus has relatively high biomass yield, calorific value, and cellulose content compared with other lignocellulosic bioenergy crops, which make it one of the most promising feedstocks for the production of second‐generation biofuels. Moreover, Miscanthus can endure soil pollutions caused by various heavy metals and survive in a variety of adverse environmental conditions. Therefore, it also has potential applications in ecological remediation of contaminated soil, and reclamation of polluted soil and water resources. Nevertheless, more endeavors are still needed in the genetic improvement and elite cultivar breeding, large‐scale cultivation on marginal land, and efficient conversion to biofuels, when utilizing Miscanthus as a bioenergy crop. Furthermore, more efforts should also be undertaken to translate Miscanthus into a bioenergy crop with the phytoremediation potential.     

芒草中总黄酮的提取工艺研究及其挥发油成分的鉴定

以黄酮提取率为指标,采用超声辅助法研究了芒草中黄酮类化合物的提取工艺。最佳提取条件为:乙醇浓度70%、料液比1∶30、提取温度70℃、提取时间40 min、提取级数3次,在此条件下测得南荻中总黄酮含量为7.24 mg/g,并在此条件下测定了芒草10个品系中总黄酮含量为4.16~7.84 mg/g,且在不同品系间的差异均达到极显著水平。采用气质联用技术鉴定奇岗六倍体、‘GMG121’、芒杂中的挥发油成分,从中共鉴定出28种成分。本研究对芒属植物的遗传育种以及开发和综合利用具有重要意义。     

Rapid selection and identification of Miscanthus genotypes with enhanced glucan and xylan yields from hydrothermal pretreatment followed by enzymatic hydrolysis

ABSTRACT: BACKGROUND: Because many Miscanthus genotypes can be cultivated with relatively high productivity and carbohydrate content, Miscanthus has great potential as an energy crop that can support large scale biological production of biofuels. RESULTS: In this study, batch hydrothermal pretreatment at 180 °C for 35 min followed by enzymatic hydrolysis was shown to give the highest total sugar yields for Miscanthus x giganteus cv. Illinois planted in Illinois. High throughput pretreatment at 180 °C for 35 min and 17.5 min followed by co-hydrolysis in a multi-well batch reactor identified two varieties out of 80 that had significantly higher sugar yields from pretreatment and enzymatic hydrolysis than others. The differences in performance were then related to compositions of the 80 varieties to provide insights into desirable traits for Miscanthus that enhance sugar yields. CONCLUSIONS: High throughput pretreatment and co-hydrolysis (HTPH) rapidly identified promising genotypes from a wide range of Miscanthus genotypes, including hybrids of Miscanthus sacchariflorus/M. sinensis and Miscanthus lutarioriparius, differentiating the more commercially promising species from the rest. The total glucan plus xylan content in Miscanthus appeared to influence both mass and theoretical yields, while lignin and ash contents did not have a predictable influence on performance.     

Environmental Tolerances of Miscanthus sinensis in Invasive and Native Populations

Miscanthus sinensis is a moderately invasive ornamental grass species being considered as a bioenergy species in the USA and elsewhere. In this study, we show the range of environmental conditions tolerated by this species in wild populations in the USA and in Japan. Six naturalized populations in the USA and five native populations in Japan were sampled in summer 2009. In each population, environmental factors (canopy cover and soil fertility) were measured, along with measurements of size and morphology for 30 plants. Relationships between M. sinensis size and environmental variables in the two countries were determined using linear mixed effects models. Results indicated that M. sinensis can tolerate extremely wide variation in soil and climate conditions in the populations we sampled across both ranges, suggesting that it could be successfully grown across a wide distribution in the USA, both intentionally as a bioenergy crop and unintentionally as an escaped invader. Plant size generally responded to different environmental conditions in both ranges, with USA plants being negatively influenced by canopy cover and Japanese plants being positively influenced by soil fertility measures. We recommend caution in growing M. sinensis for bioenergy or ornamental purposes to minimize escape outside of its native range.     

Distribution, morphological description, and molecular characterization of Xiphinema and Longidorous spp. associated with plants (Miscanthus spp. and Panicum virgatum) used for biofuels

Longidorid plant pathogens, specifically Longidorus and Xiphinema , are known to reduce the biomass and seed yields of several monocotyledonous host plants and may serve as vectors for certain plant viruses. The occurrence and densities of species in these genera in the Midwestern United States may be of concern in the production of two biofuel plant species – Miscanthus ( Miscanthus × giganteus ) and Panicum virgatum . Here we present results from a morphological and molecular survey of plant parasitic nematodes in which two species of Xiphinema ( X. americanum and X. rivesi ) and one species of Longidorus ( L. breviannulatus ) were detected. A total of 73 soil samples were collected from 19 sites (nine Miscanthus and 10 P. virgatum ) in three states during 2008. Xiphinema spp. was found in 36% of the Miscanthus plots and in 26% of the P. virgatum plots with a population density range of 22–857/100 cm³ soil. L. breviannulatus was detected in association with Miscanthus plot in Havana, IL, in large numbers (54/100 cm³ soil). The occurrence of this species was associated with severe damage to the fibrous root system, including stunting and necrosis. Short notes, measurements, illustrations and molecular data are given to the species level.     

Characterization of a genetic resource collection for Miscanthus (Saccharinae,Andropogoneae, Poaceae) using AFLP and ISSR PCR

Amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat markers were employed to characterize a genetic resource collection of Miscanthus, a grass under trial in Europe as a biomass crop. The 26 polymorphic markers produced by two ISSR fingerprinting primers were able to discriminate taxa and identify putative clones. AFLP fingerprints were fully reproducible and produced a larger number of markers for the three primer pairs tested, of which 998 were polymorphic (representing 79.3% of all bands). AFLP markers distinguished species, infra-specific taxa (varieties and cultivars) and putatively clonal material. They were also used to assess the inter-relationships of the taxa, to investigate the origin of important hybrid plants and to estimate the overall level of genetic variation in the collection. They were useful for assessing the species status of certain taxa such as M. transmorrisonensis, an endemic from Taiwan that was clearly distinct from M. sinensis; whereas other taxa of disputed species status, such as M. condensatus and M. yakushimanum were not genetically distinct from M. sinensis. The AFLP markers detected a high degree of infra-specific variation and allowed subdivisions of the genetic resource collection to be made, particularly within M. sinensis.     

Setaria viridis and Setaria italica, model genetic systems for the Panicoid grasses

Setaria italica and its wild ancestor Setaria viridis are diploid C(4) grasses with small genomes of ～515 Mb. Both species have attributes that make them attractive as model systems. Setaria italica is a grain crop widely grown in Northern China and India that is closely related to the major food and feed crops maize and sorghum. A large collection of S. italica accessions are available and thus opportunities exist for association mapping and allele mining for novel variants that will have direct application in agriculture. Setaria viridis is the weedy relative of S. italica with many attributes suitable for genetic analyses including a small stature, rapid life cycle, and prolific seed production. Setaria sp. are morphologically similar to most of the Panicoideae grasses, including major biofuel feedstocks, switchgrass (Panicum virgatum) and Miscanthus (Miscanthus giganteus). They are broadly distributed geographically and occupy diverse ecological niches. The cross-compatibility of S. italica and S. viridis also suggests that gene flow is likely between wild and domesticated accessions. In addition to serving as excellent models for C(4) photosynthesis, these grasses provide novel opportunities to study abiotic stress tolerance and as models for bioenergy feedstocks.     

Meeting US biofuel goals with less land: the potential of Miscanthus

Biofuels from crops are emerging as a Jekyll & Hyde – promoted by some as a means to offset fossil fuel emissions, denigrated by others as lacking sustainability and taking land from food crops. It is frequently asserted that plants convert only 0.1% of solar energy into biomass, therefore requiring unacceptable amounts of land for production of fuel feedstocks. The C₄ perennial grass Miscanthus × giganteus has proved a promising biomass crop in Europe, while switchgrass ( Panicum virgatum ) has been tested at several locations in N. America. Here, replicated side-by-side trials of these two crops were established for the first time along a latitudinal gradient in Illinois. Over 3 years of trials, Miscanthus × giganteus achieved average annual conversion efficiencies into harvestable biomass of 1.0% (30 t ha⁻¹) and a maximum of 2.0% (61 t ha⁻¹), with minimal agricultural inputs. The regionally adapted switchgrass variety Cave-in-Rock achieved somewhat lower yields, averaging 10 t ha⁻¹. Given that there has been little attempt to improve the agronomy and genetics of these grasses compared with the major grain crops, these efficiencies are the minimum of what may be achieved. At this 1.0% efficiency, 12 million hectares, or 9.3% of current US cropland, would be sufficient to provide 133 × 10⁹ L of ethanol, enough to offset one-fifth of the current US gasoline use. In contrast, maize grain from the same area of land would only provide 49 × 10⁹ L, while requiring much higher nitrogen and fossil energy inputs in its cultivation.     

Does greater leaf‐level photosynthesis explain the larger solar energy conversion efficiency of Miscanthus relative to switchgrass?

C₄ perennial grasses are being considered for bioenergy because of their high productivity and low inputs. In side-by-side replicated trials, Miscanthus ( Miscanthus x giganteus ) has previously been found more than twice as productive as switchgrass ( Panicum virgatum ). The hypothesis that this difference is attributable to higher leaf photosynthetic rates was tested on established plots of switchgrass and Miscanthus in central Illinois with >3300 individual measurements on 20 dates across the 2005 and 2006 growing seasons. Seasonally integrated leaf-level photosynthesis was 33% higher in Miscanthus than switchgrass ( P  < 0.0001). This increase in carbon assimilation comes at the expense of additional transpiration since stomatal conductance was on average 25% higher in Miscanthus ( P  < 0.0001). Whole-chain electron transport rate, measured simultaneously by modulated chlorophyll fluorescence, was similarly 23% higher in Miscanthus ( P  < 0.0001). Efficiencies of light energy transduction into whole chain photosynthetic electron transport, leaf nitrogen use and leaf water use were all significantly higher in Miscanthus. These may all contribute to its higher photosynthetic rates, and in turn, productivity. Systematic measurement of photosynthesis over two complete growing seasons in the field provides a unique dataset explaining why the productivity of these two species differs and for validating mechanistic production models for these emerging bioenergy crops.     

Yield‐biodiversity trade‐off in patchy fields of Miscanthus × giganteus

Increasing crop productivity to meet rising demands for food and energy, but doing so in an environmentally sustainable manner, is one of the greatest challenges for agriculture to date. In Ireland, Miscanthus × giganteus has the potential to become a major feedstock for bioenergy production, but the economic feasibility of its cultivation depends on high yields. Miscanthus fields can have a large number of gaps in crop cover, adversely impacting yield and hence economic viability. Predominantly positive effects of Miscanthus on biodiversity reported from previous research might be attributable to high crop patchiness, particularly during the establishment phase. The aim of this research was to assess crop patchiness on a field scale and to analyse the relationship between Miscanthus yield and species richness and abundance of selected taxa of farmland wildlife. For 14 Miscanthus fields at the end of their establishment phase (4–5 years after planting), which had been planted either on improved grassland (MG) or tilled arable land (MT), we determined patchiness of the crop cover, percentage light penetration (LP) to the lower canopy, Miscanthus shoot density and height, vascular plants and epigeic arthropods. Plant species richness and noncrop vegetation cover in Miscanthus fields increased with increasing patchiness, due to higher levels of LP to the lower canopy. The species richness of ground beetles and the activity density of spiders followed the increase in vegetation cover. Plant species richness and activity density of spiders on both MT and MG fields, as well as vegetation cover and activity density of ground beetles on MG fields, were negatively associated with Miscanthus yield. In conclusion, positive effects of Miscanthus on biodiversity can diminish with increasing productivity. This matter needs to be considered when assessing the relative ecological impacts of developing biomass crops in comparison with other land use.