Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sac... more Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sacchariflorus has a broad geographic range, is used to produce paper in China and is one of the parents (along with Miscanthus sinensis) of the important biomass species Miscanthus × giganteus. The largest study of M. sacchariflorus population genetics to date is reported here. Collections included 764 individuals across East Asia. Samples were genotyped with 34 605 single nucleotide polymorphisms (SNPs) derived from restriction site-associated DNA sequencing (RAD-seq) and ten plastid microsatellites, and were subjected to ploidy analysis by flow cytometry. Six major genetic groups within M. sacchariflorus were identified using SNP data: three diploid groups, comprising Yangtze (M. sacchariflorus ssp. lutarioriparius), N China and Korea/NE China/Russia; and three tetraploid groups, comprising N China/Korea/Russia, S Japan and N Japan. Miscanthus sacchariflorus ssp. lutarioriparius was deri...
To facilitate the domestication and breeding of Miscanthus for use as a bioenergy crop, it will b... more To facilitate the domestication and breeding of Miscanthus for use as a bioenergy crop, it will be necessary to develop a diverse germplasm collection from which to map and select traits of interest. We are using chloroplast microsatellites, GoldenGate SNPs, and RADseq to genetically characterize a collection of M. sinensis. This collection includes 128 accessions from Japan, 188 from South Korea, 309 from China, and 44 from naturalized populations in the United States, as well as 81 ornamental cultivars. We found two major groups of chloroplast haplotypes, one of which has a split north-south distribution in Asia. The region adjacent to the Yangtze River in China has the greatest diversity of chloroplast haplotypes. SNP markers reveal a large amount of geographic structure to the distribution of genetic diversity across the native range. Ornamental cultivars and naturalized US populations cluster with M. sinensis from Southern Japan, representing only a fraction of the genetic dive...
The genus Miscanthus is a perennial C4 grass native to eastern Asia and is a promising candidate ... more The genus Miscanthus is a perennial C4 grass native to eastern Asia and is a promising candidate bioenergy crop for cool temperate areas. Flowering time is a crucial factor governing regional and seasonal adaptation; in addition, it is also a key target trait for extending the vegetative phase to improve biomass potential. Homologues of CONSTANS (CO)/Heading date 1(Hd1) were cloned from Miscanthus sinensis and named MsiHd1. Sequences of MsiHd1 homologues were compared among 24 wild M. sinensis accessions from Japan, 14 from China, and three from South Korea. Two to five MsiHd1 alleles in each accession were identified, suggesting that MsiHd1 consists of at least three loci in the Miscanthus genome. Verifying the open reading frame in MsiHd1, they were classified as putative functional alleles without mutations or non-functional alleles caused by indels. The Neighbor-Joining tree indicated that one of the multiple MsiHd1 loci is a pseudogene locus without any functional alleles. The ...
... As the primary dominant species in a grassland community found throughout Japan (Numata, 1969... more ... As the primary dominant species in a grassland community found throughout Japan (Numata, 1969; Koyama, 1987), M. sinensis has also been used as livestock feed (Yamane, 1973; Matumura & Iwata, 1976; Otaki, 1999) and organic fertilizer (Yamane, 1973; Matumura & Iwata ...
Looming petroleum shortages and projected negative impacts of human-induced climate change may be... more Looming petroleum shortages and projected negative impacts of human-induced climate change may be partly alleviated by the development and use of bioenergy feedstock crops. Miscanthus ×giganteus, a highly productive sterile triploid hybrid grass that was discovered in Japan several decades ago, has considerable potential as an alternative source of energy. Given the risks, however, involved in the reliance upon production of one clone of this hybrid, which is a natural cross between Miscanthus sacchariflorus and Miscanthus sinensis, for lignocellulosic bioenergy production, natural occurrences of triploidy were investigated in sympatric populations of tetraploid M. sacchariflorus and diploid M. sinensis in Japan. Seeds were counted and DNA content was estimated by flow cytometry for plants of M. sacchariflorus and M. sinensis in several sympatric populations throughout Japan. Chromosomes were also counted for select plants. Based on seed-set data, M. sacchariflorus has significantly lower seed set than M. sinensis in Japan. Putative triploid seeds were found on M. sacchariflorus plants in southern Japan. This is the first report of the natural occurrence of Miscanthus triploid plants in several decades. If found to be sterile and similar in productivity to the commonly cultivated clone of M. ×giganteus, these triploid plants might serve as additional sources of genetic variation for bioenergy production. Seed set data also indicates that other triploid plants might be found in more northern regions of Japan.
Lignocellulosic components including hemicellulose, cellulose and lignin are the three major comp... more Lignocellulosic components including hemicellulose, cellulose and lignin are the three major components of plant cell walls, and their proportions in biomass crops, such as Miscanthus sinensis, greatly impact feed stock conversion to liquid fuels or bio-products. In this study, the feasibility of using visible and near infrared (VIS/NIR) spectroscopy to rapidly quantify hemicellulose, cellulose and lignin in M. sinensis was investigated. Initially, prediction models were established using partial least squares (PLS), least squares support vector machine regression (LSSVR), and radial basis function neural network (RBF_NN) based on whole wavelengths. Subsequently, 23, 25 and 27 characteristic wavelengths for hemicellulose, cellulose and lignin, respectively, were found to show significant contribution to calibration models. Three determination models were eventually built by PLS, LS-SVM and ANN based on the characteristic wavelengths. Calibration models for lignocellulosic components were successfully developed, and can now be applied to assessment of lignocellulose contents in M. sinensis.
Miscanthus is a genus of perennial C4 grasses native to East Asia. It includes the emerging ligno... more Miscanthus is a genus of perennial C4 grasses native to East Asia. It includes the emerging ligno-cellulosic biomass crop M. ×giganteus, a hybrid between M. sinensis and M. sacchariflorus Biomass yield and cold tolerance are of particular interest in Miscanthus, given that this crop is more temperate adapted than its C4 relatives maize, sorghum and sugarcane. A plant exploration was conducted in eastern Russia, at the northern extreme of the native range for Miscanthus, with collections including 174 clonal germplasm accessions (160 M. sacchariflorus and 14 M. sinensis) from 47 sites. Accessions were genotyped by restriction site-associated DNA sequencing (RAD-seq) and plastid microsatellites. Miscanthus sinensis was found in maritime climates near Vladivostok (43·6°N) and on southern Sakhalin Island (46·6°N). Miscanthus sacchariflorus was found inland at latitudes as high as 49·3°N, where M. sinensis was absent. Most M. sacchariflorus accessions were diploid, but approx. 2 % were t...
... In major perennial forage grass species, such as perennial ryegrass (Lolium perenne L.), popu... more ... In major perennial forage grass species, such as perennial ryegrass (Lolium perenne L.), populations were developed to develop molecular marker linkage maps and to identify QTL for agronomically impor-tant traits (Bert et al., 1999; Armstead et al., 2002; Jones et al., 2002). ...
ABSTRACT Fructans are water-soluble fructose oligomers and polymers that are accumulated in many ... more ABSTRACT Fructans are water-soluble fructose oligomers and polymers that are accumulated in many plants, especially the C3 temperate grasses. Fructan accumulation enhances cold and drought tolerance in grasses, and the increased soluble carbohydrates improve forage quality, especially the nutritional value animals can gain from these grasses. The C4 grasses yield high biomass but their inability to accumulate fructans coupled with their low water-soluble carbohydrate (WSC) content are limiting factors for the forage quality of these grasses. The aim of this study was to produce fructans and increase the total soluble sugar content by introducing and expressing fructan biosynthesis genes in a C4 grass, bahiagrass (Paspalum notatum Fl€ugge). Using particle bombardment, we transformed bahiagrass with a wheat-derived sucrose:sucrose 1-fructosyltransferase (1-SST) gene and a sucrose:fructan 6-fructosyltransferase (6-SFT) gene, both of which were regulated by the cauliflower mosaic virus (CaMV) 35S promoter. Transcripts of the 1-SST and 6-SFT genes were detected in six out of 12 transgenic plants carrying both genes. Transgenic bahiagrass plants expressing the 1-SST and 6-SFT genes accumulated 1-kestose and nystose. Although further polymerization of fructose units by 6-SFT activity was not detected in transgenic plants, the total soluble sugar content increased. These results suggest that transgenic bahiagrass plants expressing wheat fructosyltransferase genes created a new metabolic pathway for fructan biosynthesis, which is an important requirement for producing fructans in C4 forage grasses with the objective of increasing the total soluble sugar content of these plants. This is the first report of introducing plant-derived fructosyltransferases with the goal of increasing fructan accumulation in C4 forage grasses.
Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sac... more Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sacchariflorus has a broad geographic range, is used to produce paper in China and is one of the parents (along with Miscanthus sinensis) of the important biomass species Miscanthus × giganteus. The largest study of M. sacchariflorus population genetics to date is reported here. Collections included 764 individuals across East Asia. Samples were genotyped with 34 605 single nucleotide polymorphisms (SNPs) derived from restriction site-associated DNA sequencing (RAD-seq) and ten plastid microsatellites, and were subjected to ploidy analysis by flow cytometry. Six major genetic groups within M. sacchariflorus were identified using SNP data: three diploid groups, comprising Yangtze (M. sacchariflorus ssp. lutarioriparius), N China and Korea/NE China/Russia; and three tetraploid groups, comprising N China/Korea/Russia, S Japan and N Japan. Miscanthus sacchariflorus ssp. lutarioriparius was deri...
To facilitate the domestication and breeding of Miscanthus for use as a bioenergy crop, it will b... more To facilitate the domestication and breeding of Miscanthus for use as a bioenergy crop, it will be necessary to develop a diverse germplasm collection from which to map and select traits of interest. We are using chloroplast microsatellites, GoldenGate SNPs, and RADseq to genetically characterize a collection of M. sinensis. This collection includes 128 accessions from Japan, 188 from South Korea, 309 from China, and 44 from naturalized populations in the United States, as well as 81 ornamental cultivars. We found two major groups of chloroplast haplotypes, one of which has a split north-south distribution in Asia. The region adjacent to the Yangtze River in China has the greatest diversity of chloroplast haplotypes. SNP markers reveal a large amount of geographic structure to the distribution of genetic diversity across the native range. Ornamental cultivars and naturalized US populations cluster with M. sinensis from Southern Japan, representing only a fraction of the genetic dive...
The genus Miscanthus is a perennial C4 grass native to eastern Asia and is a promising candidate ... more The genus Miscanthus is a perennial C4 grass native to eastern Asia and is a promising candidate bioenergy crop for cool temperate areas. Flowering time is a crucial factor governing regional and seasonal adaptation; in addition, it is also a key target trait for extending the vegetative phase to improve biomass potential. Homologues of CONSTANS (CO)/Heading date 1(Hd1) were cloned from Miscanthus sinensis and named MsiHd1. Sequences of MsiHd1 homologues were compared among 24 wild M. sinensis accessions from Japan, 14 from China, and three from South Korea. Two to five MsiHd1 alleles in each accession were identified, suggesting that MsiHd1 consists of at least three loci in the Miscanthus genome. Verifying the open reading frame in MsiHd1, they were classified as putative functional alleles without mutations or non-functional alleles caused by indels. The Neighbor-Joining tree indicated that one of the multiple MsiHd1 loci is a pseudogene locus without any functional alleles. The ...
... As the primary dominant species in a grassland community found throughout Japan (Numata, 1969... more ... As the primary dominant species in a grassland community found throughout Japan (Numata, 1969; Koyama, 1987), M. sinensis has also been used as livestock feed (Yamane, 1973; Matumura & Iwata, 1976; Otaki, 1999) and organic fertilizer (Yamane, 1973; Matumura & Iwata ...
Looming petroleum shortages and projected negative impacts of human-induced climate change may be... more Looming petroleum shortages and projected negative impacts of human-induced climate change may be partly alleviated by the development and use of bioenergy feedstock crops. Miscanthus ×giganteus, a highly productive sterile triploid hybrid grass that was discovered in Japan several decades ago, has considerable potential as an alternative source of energy. Given the risks, however, involved in the reliance upon production of one clone of this hybrid, which is a natural cross between Miscanthus sacchariflorus and Miscanthus sinensis, for lignocellulosic bioenergy production, natural occurrences of triploidy were investigated in sympatric populations of tetraploid M. sacchariflorus and diploid M. sinensis in Japan. Seeds were counted and DNA content was estimated by flow cytometry for plants of M. sacchariflorus and M. sinensis in several sympatric populations throughout Japan. Chromosomes were also counted for select plants. Based on seed-set data, M. sacchariflorus has significantly lower seed set than M. sinensis in Japan. Putative triploid seeds were found on M. sacchariflorus plants in southern Japan. This is the first report of the natural occurrence of Miscanthus triploid plants in several decades. If found to be sterile and similar in productivity to the commonly cultivated clone of M. ×giganteus, these triploid plants might serve as additional sources of genetic variation for bioenergy production. Seed set data also indicates that other triploid plants might be found in more northern regions of Japan.
Lignocellulosic components including hemicellulose, cellulose and lignin are the three major comp... more Lignocellulosic components including hemicellulose, cellulose and lignin are the three major components of plant cell walls, and their proportions in biomass crops, such as Miscanthus sinensis, greatly impact feed stock conversion to liquid fuels or bio-products. In this study, the feasibility of using visible and near infrared (VIS/NIR) spectroscopy to rapidly quantify hemicellulose, cellulose and lignin in M. sinensis was investigated. Initially, prediction models were established using partial least squares (PLS), least squares support vector machine regression (LSSVR), and radial basis function neural network (RBF_NN) based on whole wavelengths. Subsequently, 23, 25 and 27 characteristic wavelengths for hemicellulose, cellulose and lignin, respectively, were found to show significant contribution to calibration models. Three determination models were eventually built by PLS, LS-SVM and ANN based on the characteristic wavelengths. Calibration models for lignocellulosic components were successfully developed, and can now be applied to assessment of lignocellulose contents in M. sinensis.
Miscanthus is a genus of perennial C4 grasses native to East Asia. It includes the emerging ligno... more Miscanthus is a genus of perennial C4 grasses native to East Asia. It includes the emerging ligno-cellulosic biomass crop M. ×giganteus, a hybrid between M. sinensis and M. sacchariflorus Biomass yield and cold tolerance are of particular interest in Miscanthus, given that this crop is more temperate adapted than its C4 relatives maize, sorghum and sugarcane. A plant exploration was conducted in eastern Russia, at the northern extreme of the native range for Miscanthus, with collections including 174 clonal germplasm accessions (160 M. sacchariflorus and 14 M. sinensis) from 47 sites. Accessions were genotyped by restriction site-associated DNA sequencing (RAD-seq) and plastid microsatellites. Miscanthus sinensis was found in maritime climates near Vladivostok (43·6°N) and on southern Sakhalin Island (46·6°N). Miscanthus sacchariflorus was found inland at latitudes as high as 49·3°N, where M. sinensis was absent. Most M. sacchariflorus accessions were diploid, but approx. 2 % were t...
... In major perennial forage grass species, such as perennial ryegrass (Lolium perenne L.), popu... more ... In major perennial forage grass species, such as perennial ryegrass (Lolium perenne L.), populations were developed to develop molecular marker linkage maps and to identify QTL for agronomically impor-tant traits (Bert et al., 1999; Armstead et al., 2002; Jones et al., 2002). ...
ABSTRACT Fructans are water-soluble fructose oligomers and polymers that are accumulated in many ... more ABSTRACT Fructans are water-soluble fructose oligomers and polymers that are accumulated in many plants, especially the C3 temperate grasses. Fructan accumulation enhances cold and drought tolerance in grasses, and the increased soluble carbohydrates improve forage quality, especially the nutritional value animals can gain from these grasses. The C4 grasses yield high biomass but their inability to accumulate fructans coupled with their low water-soluble carbohydrate (WSC) content are limiting factors for the forage quality of these grasses. The aim of this study was to produce fructans and increase the total soluble sugar content by introducing and expressing fructan biosynthesis genes in a C4 grass, bahiagrass (Paspalum notatum Fl€ugge). Using particle bombardment, we transformed bahiagrass with a wheat-derived sucrose:sucrose 1-fructosyltransferase (1-SST) gene and a sucrose:fructan 6-fructosyltransferase (6-SFT) gene, both of which were regulated by the cauliflower mosaic virus (CaMV) 35S promoter. Transcripts of the 1-SST and 6-SFT genes were detected in six out of 12 transgenic plants carrying both genes. Transgenic bahiagrass plants expressing the 1-SST and 6-SFT genes accumulated 1-kestose and nystose. Although further polymerization of fructose units by 6-SFT activity was not detected in transgenic plants, the total soluble sugar content increased. These results suggest that transgenic bahiagrass plants expressing wheat fructosyltransferase genes created a new metabolic pathway for fructan biosynthesis, which is an important requirement for producing fructans in C4 forage grasses with the objective of increasing the total soluble sugar content of these plants. This is the first report of introducing plant-derived fructosyltransferases with the goal of increasing fructan accumulation in C4 forage grasses.
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