Giganteus Plants Research Articles

An efficient system for high-quality large-scale micropropagation of Miscanthus × giganteus plants

Miscanthus × giganteus is a perennial grass considered to be one of the most promising biofuel and bioenergy crops because of its high biomass and quality, and low requirement for fertilizers and pesticides. Market demand for Miscanthus is rapidly increasing. However, M. × giganteus is a triploid that cannot produce viable seeds, and it has traditionally been propagated through rhizome division, which is low throughput and labor-intensive. Plant tissue culture provides the potential to propagate M. × giganteus in vitro while maintaining the original plant characteristics. Although protocols exist for M. × giganteus micropropagation, the multiplication rate and plant quality need to be improved to meet commercial demands. For this research, we have assessed callus induction, callus multiplication, plantlet regeneration, shoot multiplication, shoot quality improvement, rooting, plant acclimatization, and survival in the greenhouse and in the field. Through these studies, we have developed an efficient system for high-quality, large-scale micropropagation of M. × giganteus. The plants produced from our protocol exhibited more than 99% survival in soil due to the production of vigorous shoots and roots.

Optimization of Enzymatic Hydrolysis Process of Mischantus giganteus Plants, which were Physical/Chemical Pretreated to Obtain Biogas

Biogas plays an important role, with a very high potential for development. To convert biomass into biogas can be used different types of waste: waste and by-products from agriculture, agro-industry and food industry, and household and generally the waste from a lot of everyday activities of society. In principle, the bioconversion of lignocellulosic biomass produced useful and valuable is done in several stages: (i) preliminary pretreatment (mechanical, physical or chemical) to reduce non starchy macromolecules (eg. lignin);(ii) enzymatic hydrolysis of cellulose and hemicellulose fermenting carbohydrates (mainly glucose and xylose);(iii) the use of these molecules either microbial growth or for the synthesis of nano-type compounds;(iv) separation and purification of the final product. The aim of the paper is to determine pe characteristics of pretreatment with H2SO4, to obtain biogas from Mischantus giganteus plants. The best results were obtained in chemically treated variants B with H2SO4 concentration greater than 1%; when using the enzyme MethaPlus L 100 same trend is observed for variants with advanced hydration over 1.0% H2SO4. The examination of variants with more than 1.0% H2SO4 is apparent that the rate of hydrolysis differ very little in terms of effectiveness and safety equipment used in obtaining biogas is preferably choosing 1.0% H2SO4.

Evaluating silicon concentrations in biofuel feedstock crops Miscanthus and switchgrass

Abstract Silicon (Si) concentrations in biofuel feedstock crops have a critical role in combustion processes. The purpose of this study was to quantify Si concentrations in plant biomass samples and to evaluate the contributing factors for Si concentrations. We determined total Si concentrations in Miscanthus × giganteus ( M . × giganteus ) collected from various research trial plots in the eastern U.S. and in Miscanthus spp. and Panicum virgatum , ‘Cave-in-Rock’ (switchgrass) from an additional eight trial plots established across Illinois. Whole aboveground plant biomass at each site were air-dried and ground. Total Si concentrations in plant samples were determined by dry-ashing plant tissue in a muffle furnace, followed by alkaline fusion and then colorimetric analysis. Average Si concentrations in statewide M . × giganteus plant samples ranged from 0.72% to 1.6% and samples from within Illinois ranged from 0.55% to 2.4%. The overall median value of concentrations in M . × giganteus samples among all sites was 1.08%. The median value in switchgrass samples (1.5%) was 1.4 times higher than that for M . × giganteus . Among six other Miscanthus spp. samples from the Urbana trial plot in Illinois, Si concentrations were about 1/3 that of M . × giganteus . Variation in Si concentrations tended to be associated with temperature and precipitation of the location where the biofuel crops are being grown. We did not find any relationship between soil type and plant Si concentrations. Long-term evaluations of soil mineral concentrations and additional environmental factors are required to better understand the contributing factors for Si concentrations.

Plant age and in vitro or in vivo propagation considerably affect cold tolerance of Miscanthus × giganteus

The aims of the study were to determine why young Miscanthus × giganteus plants are more frost sensitive during the first winter than older plants, to compare cold tolerance of plants propagated in in vivo and in vitro conditions, and to select plants with higher cold tolerance. The study was performed in three experiments in which plants were prehardened at 12 °C for 2 weeks, hardened at 5 °C for 3 weeks and next chilled at 0 °C or −3 °C for 3 or 14 days. Afterwards shoot regrowth from rhizomes was evaluated. In Experiment 1 frost tolerance of young plants obtained from a horticultural farm and plants that had already survived the first winter in the field was compared on the basis of LT 50 coefficient. In Experiment 2 frost tolerance of plants obtained in vivo and in vitro was compared. Experiment 3 was performed on four groups of plants: in vivo and in vitro obtained plants which were twice selected in cold, as well as in vivo and in vitro obtained plants which were cold treated once. Plants of all these groups were analysed with respect to their frost tolerance. They were prehardened, hardened and subjected to a temperature of 0 °C or −3 °C for 14 days. The changes in processes accompanying cold acclimation occurring in the rhizomes or leaves of these plants were investigated. The content of abscisic acid, low-molecular antioxidants and phenolics, as well as catalase and non-specific peroxidase activities were analysed. Young commercially obtained plants were more frost sensitive than plants which had survived the first winter in the field. This effect could be caused by a small amount of storage compounds accumulated in finely divided rhizomes produced in a horticultural farm. Prehardening temperature of 12 °C caused more considerable changes in cold acclimation processes in Miscanthus rhizomes than hardening temperature of 5 °C. Plants propagated by in vitro culture were more cold tolerant but only in the first vegetative season compared to plants obtained in vivo. Plants chilled twice demonstrated a higher low-molecular antioxidant level, as well as a greater capability of phenolic accumulation compared to plants which were once cold stressed. Regardless of the recurrence of cold acclimation, ABA level was significantly increased in leaves by prehardening and in rhizomes by hardening. Each repetition of cold acclimation increased cold tolerance and shoot regeneration ability of M. × giganteus rhizomes.

First Report of Pithomyces chartarum Causing a Leaf Blight of Miscanthus × giganteus in Kentucky.

Miscanthus × giganteus is a warm-season perennial grass, native to eastern Asia. Brought into the United States as a landscape plant, it is currently being considered as a potential biomass fuel crop. In August 2009, a newly established and a 2-year-old M. × giganteus field research trial near Lexington, KY were found to have 100% incidence of severe leaf blight. Brown, mosaic-like, coalesced necrotic lesions covered leaf blades and sheaths on every stand, ultimately killing some leaves and tillers. The disease was more destructive in the newly established trial where 4- to 5-month-old M. × giganteus tillers were killed. No fruiting bodies were found immediately on diseased leaves. However, surface-disinfested diseased leaf tissue produced a sooty black mass of conidia after 1 week following incubation in a petri dish moisture chamber at 25°C in the dark. Single conidia isolations were made on half-strength potato dextrose agar (HSPDA) amended with 25 mg/liter of rifamycin and incubated at 25°C. Morphological characteristics of the fungus fit those originally described for Pithomyces chartarum (Berk. & Curt.) M.B. Ellis (2). Colonies were fast growing on HSPDA, at first hyaline, then shortly punctiform, grayish black, up to 1-mm diameter, and then became confluent, producing several dark brown multicellular conidia on small peg-like denticles on branched conidiophores. Every detached conidium had a small piece of the denticle attached to its base. The conidia were echinulate, broadly ellipsoidal, pyriform, 18 to 29 × 11 to 18 μm, with three transverse septa, and a longitudinal septum constricted at the transverse septa. The identity of the fungus was confirmed by sequence analysis of the internal transcribed spacers (ITS) region of the nuclear ribosomal DNA. The 615-bp cloned and sequenced amplicon (Accession No. GU195649) was 99% identical to sequences from multiple isolates of Leptosphaerulina chartarum (anamorph Pithomyces chartarum) in the GenBank. Five potted M. × giganteus plants (45 days old) were spray inoculated with an aqueous conidial suspension (2 × 106 conidia/ml) and incubated in one tier of a two-tiered-growth chamber at 86 to 90% relative humidity. Initial incubation was in the dark at 26°C for 48 h, and thereafter at alternating 15 h of light (320 μmol) at 25°C and 9 h of darkness at 23°C. Control plants were sprayed with sterile water and incubated in the second tier of the same growth chamber. A week after inoculation, leaf blight developed on all inoculated plants, but not the controls. P. chartarum was reisolated from infected leaves 2 weeks after inoculation. To our knowledge, this is the first report of P. chartarum causing a disease on Miscanthus (3). The fungus is cosmopolitan, usually saprophytic, but can cause diseases on a wide range of plants as well as produce mycotoxins (3). It has been reported to cause a leaf spot of smooth bromegrass (Bromus inermis) in Nebraska (1) and a leaf blight of wheat (Triticum aestivum) in Hungary (4). The observed disease severity suggests P. chartarum could potentially limit M. × giganteus production as an ethanol feedstock.

First Report of Clonostachys rhizophaga as a Pathogen of Dendrocalamus giganteus in Mozambique.

Dendrocalamus giganteus Wall. ex Munro, a wild plant belonging to the family Poaceae, is widespread in Mozambique where it is used as a construction material. At the end of 2007, disease symptoms have been observed on D. giganteus plants growing in the neighborhood of Maputo. Diseased plants showed longitudinal dark streaks on the stem surface to which corresponded internal vascular browning and chlorosis in wide leaves that gradually developed into necrosis. At the final stage of the disease plants died. To isolate the pathogen, stem segments collected during September 2008 were surface sterilized with 1% HgCl2 for 30 s, rinsed with sterile deionized water for 30 s, and incubated on potato dextrose agar (PDA) medium at 22°C in the dark for 2 weeks. Monosporic cultures of the isolated fungus formed dimorphic Verticillium-like (primary) or penicillate (secondary) conidiophores and ovoidal to elongate, minutely curved, hyaline conidia, 5 to 9.5 × 2.5 to 4.5 μm, with laterally displaced hilum. These characteristics are typical of Clonostachys rhizophaga Schroers (3). Identification was confirmed by the Centraalbureau voor Schimmelcultures (Utrecht, the Netherlands) on the basis of the β-tubulin (tub2) gene sequence (3). For our isolate CBS 125416, the tub2 sequence was 100% similar to that of the C. rizhophaga strain CBS 124511 (GenBank Accession No. FJ 593883) (1). To verify the pathogenicity of the fungus, a 5-mm-diameter mycelial plug obtained from 2-week-old colonies grown on PDA was affixed to a portion of the stem of D. giganteus from which the superficial tissues had been removed and the inoculation site was covered with wet cotton and wrapped with Parafilm. Control plants were treated by the same method but using PDA plugs without mycelium. Twenty plants were used, ten of which were controls. They were grown in a controlled climatic chamber at 22°C with a photoperiod of 16 h at 40 μE·m-2·s-1. Two months after inoculation, all plants showed a dark area surrounded by an idropic halo on the stem surface and internal browning, whereas control plants remained healthy. C. rhizophaga was recovered from all infected plants. C. rhizophaga is apparently rare. The fungus (as Verticillium rhizophagum Tehon & Jacobs, nom. invalid.) has been previously reported from the United States, Chile, and Ecuador (4) and as a culture contaminant in Switzerland (3). More recently C. rhizophaga has been found to be associated with Pinus canariensis in Argentina (2) and it has been reported as a causal agent of chickpea wilt in Syria (1). To our knowledge, this is the first report of C. rhizophaga for subsaharian Africa. It may be under reported and more common than has been thought because of the difficulty in identifying Clonostachys species, but with the ability to identify species using tub2 (3), there can be no doubt of its identity.

Chromosome doubling of the bioenergy crop, Miscanthus×giganteus

AbstractThe 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.

Genome Size of Three Miscanthus 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.

The potential of foliar treatments for enhanced phytoextraction of heavy metals from contaminated soil

AbstractPhytoextraction is the plant‐based removal of inorganic contaminants from the soil by root absorption and subsequent translocation to harvestable plant parts. The efficiency of this technique is limited by the phytoavailability of these contaminants in the soil and the root‐to‐shoot transport. To enhance the phytoextraction efficiency, the use of soil amendments has been widely investigated. Potential risks such as increased ecotoxicological effects and leaching of mobilized contaminants caution against the use of persistent mobilizing agents. The potential use of foliar amendments to enhance mineral absorption and/or translocation offers prospects for complementing or substituting soil amendments for enhanced phytoextraction purposes. This study presents an explorative screening to evaluate the feasibility of this approach. Helianthus annuus giganteus plants were grown in moderately contaminated dredged sediment for ten weeks, with daily foliar treatments from week six onward: (1) distilled water (control), (2) magnesium‐ethylenediaminetetraacetic acid (Mg‐EDTA), (3) diethylenetriaminepentaacetic acid (DTPA), (4) nitrilotriacetic acid (NTA), (5) citric acid, (6) oxalic acid, (7) calcium acetate, (8) ammonium thiocyanate (NH4SCN), and (9) cystein. Applied doses varied between treatments based on plant tolerance: 15 μmol per plant for cystein, 60 μmol per plant for DTPA and NH4SCN, 180 μmol per plant for Mg‐EDTA and NTA and 400 μmol per plant for the organic acids. At the end of the experiment, shoot accumulation of cadmium, chromium, copper, iron, manganese, nickel, lead, and zinc was evaluated. © 2004 Wiley Periodicals, Inc.

Propagation method as an important factor in the growth and development of Miscanthus×giganteus

The morphological development of Miscanthus×giganteus plants propagated either by rhizome division or by micropropagation was studied in two field and two pot trials. Significant differences were found for the thickness of the rhizome branches as well as the number and strength of the shoots. Rhizome propagated plants had a lower number but stronger shoots and thicker rhizome branches than micropropagated plants. In the field those plants micropropagated directly by in vitro tillering developed more slowly than those micropropagated through somatic embryogenesis, both in terms of total dry matter (DM) and share of the rhizome of total DM. The analysis of the chemical composition of shoots and rhizomes showed a tendency of higher N, P, K and sugar contents in rhizomes from rhizome propagated plants. Morphological differences between propagation variants were found to last until the third ratoon although these differences diminish with increasing age of the plantation. Directly micropropagated plants have thinner stems, making them more susceptible to lodging. As no chilling occurred in any of the trials, no immediate conclusions can be drawn about differences in chilling resistance between the propagation variants.