Arvid Boe

A multiple species approach to biomass production from native herbaceous perennial feedstocks

Due to the rapid rate of worldwide consumption of nonrenewable fossil fuels, production of biofuels from cellulosic sources is receiving increased research emphasis. Here, we review the feasibility to produce lignocellulosic biomass on marginal lands that are not well-suited for conventional crop production. Large areas of these marginal lands are located in the central prairies of North America once dominated by tallgrass species. In this article, we review the existing literature, current work, and potential of two native species of the tallgrass prairie, prairie cordgrass (Spartina pectinata), and little bluestem (Schizachyrium scoparium) as candidates for commercial production of biofuel. Based on the existing literature, we discuss the need to accelerate research in the areas of agronomy, breeding, genetics, and potential pathogens. Cropping systems based on maintaining biodiversity across landscapes are essential for a sustainable production and to mitigate impact of pathogens and pests.

Morphology and biomass production of prairie cordgrass on marginal lands

Prairie cordgrass (Spartina pectinata Link.) is indigenous throughout most of the continental United States and Canada to 60°N latitude and is well suited to marginal land too wet for maize (Zea mays L.) and switchgrass (Panicum virgatum L.). Evaluations of prairie cordgrass in Europe and North America indicated it has high potential for biomass production, relative to switchgrass, in short‐season areas. Our objective was to describe morphology and biomass production and partitioning in mature stands of ‘Red River’ prairie cordgrass and determine biomass production of natural populations on marginal land. This study was conducted from 2000 to 2008 in eastern South Dakota. Mean biomass production of mature stands of Red River was 12.7 Mg ha−1. Leaves composed >88% of the biomass, and 60% of the tillers had no internodes. Belowground biomass to a depth of approximately 25 cm, not including roots, was 21 Mg ha−1. Tiller density ranged from 683 tillers m−2 for a 10‐year‐old stand to 1140 tillers m−2 for a 4‐year‐old stand. The proaxis was composed of about eight phytomers, with rhizomes originating at proximal nodes and erect tillers at distal nodes. Vegetative propagation was achieved by both phalanx and guerilla growth. Differences among natural populations for biomass were expressed on gravelly marginal land. However, production, averaged across populations, was low (1.37 Mg ha−1) and comparable to ‘Cave‐In‐Rock’ switchgrass (1.67 Mg ha−1) over a 4‐year period. The large carbon storage capacity of prairie cordgrass in proaxes and rhizomes makes it useful for carbon sequestration purposes. Prairie cordgrass should be compared with switchgrass and other C4 perennial grasses along environmental gradients to determine optimum landscape positions for each and to maximize bioenergy production and minimize inputs.

Symptoms, Distribution and Abundance of the Stem-Boring Caterpillar, Blastobasis repartella (Dietz), in Switchgrass

A potential pest of switchgrass, Panicum virgatum L., was first detected in South Dakota in 2004, where death of partially emerged leaves was noted in a small proportion of tillers. Similar “dead heart” symptoms were observed in switchgrass in Illinois during 2008 and adults of a stem-boring caterpillar were collected and identified as Blastobasis repartella (Dietz). In 2009, a survey of the central United States was used to estimate the distribution and abundance of this insect. In eight northern states, B. repartella was consistently found in both cultivated plots and natural stands of switchgrass. In four southern states, B. repartella was not detected. However, because symptoms are conspicuous for a short period of time, failure to collect stem-borers on one survey date for each southern location does not necessarily define the limit of distribution for B. repartella. Sampling in four northern states showed the proportion of tillers damaged by B. repartella ranged from 1.0–7.2%. Unlike some caterpillars that feed on native grasses, it appears that the egg-laying behavior of adult moths may preclude the use of prescribed burns as an effective method to suppress this stem-boring caterpillar. As a potential pest of switchgrass planted for biomass production, near-term research needs include refining the geographic distribution of B. repartella, quantifying potential losses of switchgrass biomass, and determining whether switchgrass may be bred for resistance this and other stem-boring insects.

Biomass and seed yields of big bluestem, switchgrass, and intermediate wheatgrass in response to manure and harvest timing at two topographic positions

A principle attribute of perennial grasses for biomass energy is the potential for high yields on marginal lands. Objectives of this study were to compare biomass and seed production of intermediate wheatgrass (Thinopyrum intermedium [Host] Barkworth and D.R. Dewey), big bluestem (Andropogon gerardii Vitman), and switchgrass (Panicum virgatum L.) as affected by harvest timing and manure application on two topographic positions (footslope and backslope). Footslope is the hillslope position that forms the inclined surface at the base of a slope and backslope forms the steepest, middle position of the hillslope. Grasses were harvested for biomass at anthesis (summer), after a killing frost (autumn), or the following spring after overwintering in the field. Seed was harvested at maturity during 2003 and 2004. Two rates of beef cattle (Bos taurus L.) manure (target rates of 0 and 150 kg total‐N ha−1) were surface applied annually. Maximum annual biomass yield ranged from 4.4 to 5.2, 2.7 to 4.2, and 3.7 to 5.6 Mg ha−1 for intermediate wheatgrass, big bluestem, and switchgrass, respectively. Biomass yields were not different between fall and spring harvest treatments. Biomass yields of big bluestem and switchgrass at the backslope position were 86% and 96% of biomass yields at the footslope position with normal precipitation, respectively. Manure application increased biomass yield approximately 30% during the second year on both topographic positions. The highest seed yield was obtained from intermediate wheatgrass, followed by switchgrass and big bluestem. Utilizing these management practices in our environment, it appears that switchgrass and big bluestem could be allowed to overwinter in the field without suffering appreciable loss of biomass.

Switchgrass, Big Bluestem, and Indiangrass Monocultures and Their Two- and Three-Way Mixtures for Bioenergy in the Northern Great Plains

High yielding, native warm-season grasses could be used as renewable bioenergy feedstocks. The objectives of this study were to determine the effect of warm season grass monocultures and mixtures on yield and chemical characteristics of harvested biomass and to evaluate the effect of initial seeding mixture on botanical composition over time. Switchgrass (Panicum virgatum L.), indiangrass [Sorghastrum nutans (L.) Nash], and big bluestem (Andropogon gerardii Vitman) were planted as monocultures and in all possible two- and three-way mixtures at three USA locations (Brookings and Pierre, SD and Morris, MN) during May 2002. Biomass at each location was harvested after a killing frost once annually from 2003 to 2005. Total biomass yield significantly increased with year at all locations. Switchgrass monocultures or mixtures containing switchgrass generally out-yielded big bluestem or indiangrass in monocultures or the binary mixture. Cellulose and hemicellulose concentrations were higher in 2004 and 2005 compared with 2003. Switchgrass or mixtures containing switchgrass tended to have less cellulose than either big bluestem or indiangrass. Results were more variable for total N, lignin, and ash. Switchgrass was the dominant component of all mixtures in which it was present while big bluestem was dominant when mixed with indiangrass. Indiangrass was maintained only in monocultures and declined over years when grown in mixtures at all locations. Our results indicated if biomass yield in the northern Great Plains is a primary objective, switchgrass should be a component of binary or tertiary mixtures that also contain big bluestem and/or indiangrass.

A New Species of Gall Midge (Diptera: Cecidomyiidae) Infesting Switchgrass in the Northern Great Plains

A new species of gall midge, Chilophaga virgati Gagné (Diptera: Cecidomyiidae), collected from tillers of switchgrass at Brookings, South Dakota (44.31134° N, 96.78374° W) is described here. Plant morphological symptoms of infestation and frequency of tillers infested are also provided. Full-grown larvae of C. virgati were found inside the sheath of the flag leaf of reproductive tillers of clonally replicated spaced plants of a selected southern upland population of switchgrass during October 2008 and 2009. Infested tillers were shorter and lighter than normal tillers and had panicles that were partially encased in the sheath of the flag leaf due to reduced elongation of the peduncle as a result of the feeding of larvae of C. virgati at the proximal end of the panicle and in the intercalary meristem area of the peduncle. Variation was found among 10 genotypes for percentage of tillers infested by C. virgati, with a range from 7.2% to 21.8%. No difference was found between years for infestation rate (12.7% in 2008 and 14.3% in 2009). The mass of infested tillers was 35% that of normal tillers, and infested tillers produced no appreciable amount of viable seeds. Results of this research revealed that C. virgati had direct negative impacts on biomass and seed production in spaced plant nurseries of switchgrass. C. virgati was also observed in seeded swards of northern upland switchgrass cultivars, but its impact in seeded swards has not yet been determined.

Investigation of the Transcriptome of Prairie Cord Grass, a New Cellulosic Biomass Crop

Prairie cordgrass (Spartina pectinata Bosc ex Link) is being developed as a cellulosic biomass crop. Development of this species will require numerous steps, including breeding, agronomy, and characterization of the species genome. The research in this paper describes the first investigation of the transcriptome of prairie cordgrass via Next Generation Sequencing Technology, 454 GS FLX. A total of 556,198 expressed sequence tags (ESTs) were produced from four prairie cordgrass tissues: roots, rhizomes, immature inflorescence, and hooks. These ESTs were assembled into 26,302 contigs and 71,103 singletons. From these data were identified, EST–SSR (simple sequence repeat) regions and cell wall biosynthetic pathway genes suitable for the development of molecular markers which can aid the breeding process of prairie cordgrass by means of marker assisted selection.

Proteomic Responses of Switchgrass and Prairie Cordgrass to Senescence

Senescence in biofuel grasses is a critical issue because early senescence decreases potential biomass production by limiting aerial growth and development. 2-Dimensional, differential in-gel electrophoresis (2D-DIGE) followed by mass spectrometry of selected protein spots was used to evaluate differences between leaf proteomes of early (ES)- and late- senescing (LS) genotypes of Prairie cordgrass (ES/LS PCG) and switchgrass (ES/LS SG), just before and after senescence was initiated. Analysis of the manually filtered and statistically evaluated data indicated that 69 proteins were significantly differentially abundant across all comparisons, and a majority (41%) were associated with photosynthetic processes as determined by gene ontology analysis. Ten proteins were found in common between PCG and SG, and nine and 18 proteins were unique to PCG and SG respectively. Five of the 10 differentially abundant spots common to both species were increased in abundance, and five were decreased in abundance. Leaf proteomes of the LS genotypes of both grasses analyzed before senescence contained significantly higher abundances of a 14-3-3 like protein and a glutathione-S-transferase protein when compared to the ES genotypes, suggesting differential cellular metabolism in the LS vs. the ES genotypes. The higher abundance of 14-3-3 like proteins may be one factor that impacts the senescence process in both LS PCG and LS SG. Aconitase dehydratase was found in greater abundance in all four genotypes after the onset of senescence, consistent with literature reports from genetic and transcriptomic studies. A Rab protein of the Ras family of G proteins and an s-adenosylmethionine synthase were more abundant in ES PCG when compared with the LS PCG. In contrast, several proteins associated with photosynthesis and carbon assimilation were detected in greater abundance in LS PCG when compared to ES PCG, suggesting that a loss of these proteins potentially contributed to the ES phenotype in PCG. Overall, this study provides important data that can be utilized toward delaying senescence in both PCG and SG, and sets a foundational base for future improvement of perennial grass germplasm for greater aerial biomass productivity.

Stand Persistence and Forage Yield of 11 Alfalfa (Medicago sativa) Populations in Semiarid Rangeland

ABSTRACT Livestock producers in the Northern Great Plains value alfalfa (Medicago sativa L.) for increasing forage production and quality in grazing lands. However, alfalfa persistence can be poor, especially under grazing. Demand exists for alfalfa that can establish and persist in semiarid grazing lands. A naturalized population of predominantly yellow-flowered alfalfa (Medicago sativa L. subsp. falcata [L.] Arcang.) was found growing and reseeding on private and public rangeland in northwestern South Dakota. This naturalized alfalfa population demonstrates persistence in this semiarid environment. A study, initiated in May 2006 at the Antelope Range and Livestock Research Station near Buffalo, South Dakota, evaluated stand persistence and forage yield of 11 alfalfa populations transplanted into mixed-grass prairie. Populations were pure falcata, predominantly falcata, hay-type sativa, or pasture-type sativa populations. Transplants were space planted on 1-m centers within three exclosures (35 × 35 m) divided into two sections, which were either mob grazed by cattle or protected from mob grazing. Mob grazing began in August 2007 and continued periodically through 2008 and 2009. Survival, plant height, plant canopy diameters, and biomass data were collected. Grazing, dry spells, and ice sheets subjected alfalfa plants to substantial stress. High mortality of grazed plants occurred during the 2008–2009 winter. Hay-type sativa and pasture-type sativa populations exposed to mob grazing had poor final survival (<19%) and forage yield in July 2010. However, pure falcata and most predominantly falcata populations had higher survival (>38%) and forage yield. Low mortality and high yield of protected plants indicated that accumulated stress from mob grazing weakened grazed plants, increasing environment-related mortality (e.g., winterkilling). Falcata-based populations persistent under mob grazing and adapted to the regional environment have potential for use in the Northern Great Plains.

Advances towards a marker-assisted selection breeding program in prairie cordgrass, a biomass crop.

Prairie cordgrass (Spartina pectinata Bosc ex Link) is an indigenous, perennial grass of North America that is being developed into a cellulosic biomass crop suitable for biofuel production. Limited research has been performed into the breeding of prairie cordgrass; this research details an initial investigation into the development of a breeding program for this species. Genomic libraries enriched for four simple sequence repeat (SSR) motifs were developed, 25 clones from each library were sequenced, identifying 70 SSR regions, and primers were developed for these regions, 35 of which were amplified under standard PCR conditions. These SSR markers were used to validate the crossing methodology of prairie cordgrass and it was found that crosses between two plants occurred without the need for emasculation. The successful cross between two clones of prairie cordgrass indicates that this species is not self-incompatible. The results from this research will be used to instigate the production of a molecular map of prairie cordgrass which can be used to incorporate marker-assisted selection (MAS) protocols into a breeding program to improve this species for cellulosic biomass production.