John E. Erickson

RNA interference suppression of lignin biosynthesis increases fermentable sugar yields for biofuel production from field-grown sugarcane

The agronomic performance, cell wall characteristics and enzymatic saccharification efficiency of transgenic sugarcane plants with modified lignin were evaluated under replicated field conditions. Caffeic acid O-methyltransferase (COMT) was stably suppressed by RNAi in the field, resulting in transcript reduction of 80%-91%. Along with COMT suppression, total lignin content was reduced by 6%-12% in different transgenic lines. Suppression of COMT also altered lignin composition by reducing syringyl units and p-coumarate incorporation into lignin. Reduction in total lignin by 6% improved saccharification efficiency by 19%-23% with no significant difference in biomass yield, plant height, stalk diameter, tiller number, total structural carbohydrates or brix value when compared with nontransgenic tissue culture-derived or transgenic control plants. Lignin reduction of 8%-12% compromised biomass yield, but increased saccharification efficiency by 28%-32% compared with control plants. Biomass from transgenic sugarcane lines that have 6%-12% less lignin requires approximately one-third of the hydrolysis time or 3- to 4-fold less enzyme to release an equal or greater amount of fermentable sugar than nontransgenic plants. Reducing the recalcitrance of lignocellulosic biomass to saccharification by modifying lignin biosynthesis is expected to greatly benefit the economic competitiveness of sugarcane as a biofuel feedstock.

Comparative Proteomic Analysis of Brassica napus in Response to Drought Stress

Drought is one of the most widespread stresses leading to retardation of plant growth and development. We examined proteome changes of an important oil seed crop, canola (Brassica napus L.), under drought stress over a 14-day period. Using iTRAQ LC-MS/MS, we identified 1976 proteins expressed during drought stress. Among them, 417 proteins showed significant changes in abundance, and 136, 244, 286, and 213 proteins were differentially expressed in the third, seventh, 10th, and 14th day of stress, respectively. Functional analysis indicated that the number of proteins associated with metabolism, protein folding and degradation, and signaling decreased, while those related to energy (photosynthesis), protein synthesis, and stress and defense increased in response to drought stress. The seventh and 10th-day profiles were similar to each other but with more post-translational modifications (PTMs) at day 10. Interestingly, 181 proteins underwent PTMs; 49 of them were differentially changed in drought-stressed plants, and 33 were observed at the 10th day. Comparison of protein expression changes with those of gene transcription showed a positive correlation in B. napus, although different patterns between transcripts and proteins were observed at each time point. Under drought stress, most protein abundance changes may be attributed to gene transcription, and PTMs clearly contribute to protein diversity and functions.

Seed train development for the fermentation of bagasse from sweet sorghum and sugarcane using a simplified fermentation process

A process was developed for seed culture expansion (3.6 million-fold) using 5% of the hemicellulose hydrolysate from dilute acid pretreatment as the sole organic nutrient and source of sugar. Hydrolysate used for seed growth was neutralized with ammonia and combined with 1.0mM sodium metabisulfite immediately before inoculation. This seed protocol was tested with phosphoric acid pretreated sugarcane and sweet sorghum bagasse using a simplified process with co-fermentation of fiber, pentoses, and hexoses in a single vessel (SScF). A 6h liquefaction (L) step improved mixing prior to inoculation. Fermentations (L+SScF process) were completed in 72 h with high yields (>80 gal/US ton). Ethanol titers for this L+SScF process ranged from 24 g/L to 32 g/L, and were limited by the bagasse concentration (10% dry matter).

Direct and indirect effects of elevated atmospheric CO2 on net ecosystem production in a Chesapeake Bay tidal wetland

The rapid increase in atmospheric CO2 concentrations (Ca ) has resulted in extensive research efforts to understand its impact on terrestrial ecosystems, especially carbon balance. Despite these efforts, there are relatively few data comparing net ecosystem exchange of CO2 between the atmosphere and the biosphere (NEE), under both ambient and elevated Ca . Here we report data on annual sums of CO2 (NEE(net) ) for 19 years on a Chesapeake Bay tidal wetland for Scirpus olneyi (C3 photosynthetic pathway)- and Spartina patens (C4 photosynthetic pathway)-dominated high marsh communities exposed to ambient and elevated Ca (ambient + 340 ppm). Our objectives were to (i) quantify effects of elevated Ca on seasonally integrated CO2 assimilation (NEE(net) = NEE(day) + NEE(night) , kg C m(-2) y(-1) ) for the two communities; and (ii) quantify effects of altered canopy N content on ecosystem photosynthesis and respiration. Across all years, NEE(net) averaged 1.9 kg m(-2) y(-1) in ambient Ca and 2.5 kg m(-2) y(-1) in elevated Ca , for the C3 -dominated community. Similarly, elevated Ca significantly (P < 0.01) increased carbon uptake in the C4 -dominated community, as NEE(net) averaged 1.5 kg m(-2) y(-1) in ambient Ca and 1.7 kg m(-2) y(-1) in elevated Ca . This resulted in an average CO2 stimulation of 32% and 13% of seasonally integrated NEE(net) for the C3 - and C4 -dominated communities, respectively. Increased NEE(day) was correlated with increased efficiencies of light and nitrogen use for net carbon assimilation under elevated Ca , while decreased NEE(night) was associated with lower canopy nitrogen content. These results suggest that rising Ca may increase carbon assimilation in both C3 - and C4 -dominated wetland communities. The challenge remains to identify the fate of the assimilated carbon.

Fermentation of sweet sorghum derived sugars to butyric acid at high titer and productivity by a moderate thermophile Clostridium thermobutyricum at 50°C.

In this study, a moderate thermophile Clostridium thermobutyricum is shown to ferment the sugars in sweet sorghum juice treated with invertase and supplemented with tryptone (10 g L(-1)) and yeast extract (10 g L(-1)) at 50°C to 44 g L(-1) butyrate at a calculated highest volumetric productivity of 1.45 g L(-1)h(-1) (molar butyrate yield of 0.85 based on sugars fermented). This volumetric productivity is among the highest reported for batch fermentations. Sugars from acid and enzyme-treated sweet sorghum bagasse were also fermented to butyrate by this organism with a molar yield of 0.81 (based on the amount of cellulose and hemicellulose). By combining the results from juice and bagasse, the calculated yield of butyric acid is approximately 90 kg per tonne of fresh sweet sorghum stalk. This study demonstrates that C. thermobutyricum can be an effective microbial biocatalyst for production of bio-based butyrate from renewable feedstocks at 50°C.

Harvest management affects biomass composition responses of C4 perennial bioenergy grasses in the humid subtropical USA

Elephantgrass (Pennisetum purpureum Schum.) and energycane (Saccharum spp. hybrid) are high‐yielding C4 grasses that are attractive biofuel feedstocks in the humid subtropics. Determining appropriate harvest management practices for optimal feedstock chemical composition is an important precursor to their successful use in production systems. In this research, we have investigated the effects of harvest timing and frequency on biomass nutrient, carbohydrate and lignin composition of UF1 and cv. Merkeron elephantgrasses and cv. L 79‐1002 energycane. Biomass properties under increased harvest frequency (twice per year) and delayed harvest (once per year after frost) were compared with a control (once per year prior to frost). There were no differences between elephantgrass entries in structural carbohydrates; however, elephantgrasses had greater structural hexose concentration than energycane for single‐harvest treatments (avg. 398 vs. 366 mg g−1), a trait that is preferred for biofuel production. Delayed harvest of energycane decreased structural hexose compared with the control (374 vs. 357 mg g−1) because nonstructural components accumulated in energycane stem as harvest was delayed. Frequent defoliation (2X) increased N, P, and ash concentrations (75% for N and P and 58% for ash) in harvested biomass compared with single‐harvest treatments. We conclude that multiple harvests per year increase the harvest period during which feedstock is available for processing, but they do not result in optimal feedstock composition. In contrast, extending the period of feedstock supply by delaying a single harvest to after first freeze did not negatively affect cell wall constituent properties, while it increased length of the harvest period by ~30 days in the southeast USA.

Biochar Decreases Atrazine and Pendimethalin Preemergence Herbicidal Activity

Abstract Biochar and vinasse are by-products of biofuel production that can be used as soil amendments. However, their addition to the soil might affect PRE herbicide activity. Although studies have shown that biochar has a high herbicide adsorption capacity, there is little information available about biochar effect on weed control especially under field conditions. Therefore, the objective of this study was to determine the influence of biochar and vinasse application on atrazine and pendimethalin availability and herbicide activity under in vitro and field conditions. In vitro atrazine and pendimethalin herbicidal activities were not influenced by vinasse addition, but biochar application reduced atrazine and pendimethalin injury for all evaluated species. A sorption experiment confirmed high affinity of biochar for atrazine and pendimethalin. Linear regression analysis showed that the slope for atrazine and pendimethalin adsorption was 16 and 4 times higher in soil with biochar than in soil alone. Under field conditions, biochar at 0.5 kg m−2 reduced atrazine and pendimethalin weed control 75% and 60%, respectively. These results suggested that the use of biochar as a soil amendment in cropping system could decrease PRE herbicide efficacy. Therefore, mitigating practices such as the use of higher rates or reliance on POST herbicides and cultivation might be necessary to ensure proper weed control. Nomenclature: Atrazine; pendimethalin; biochar; vinasse. Resumen El biochar y la vinaza son subproductos de la producción de biocombustibles que pueden ser usados como enmiendas de suelo. Sin embargo, su adición al suelo podría afectar la actividad de herbicidas PRE. Aunque estudios han mostrado que el biochar tiene una alta capacidad de adsorción de herbicidas, hay poca información disponible acerca del efecto del biochar sobre el control de malezas, especialmente bajo condiciones de campo. Por esta razón, el objetivo de este estudio fue determinar la influencia de la aplicación de biochar y de vinaza sobre la disponibilidad y actividad herbicida de atrazine y pendimethalin in vitro y en condiciones de campo. In vitro, la actividad herbicida de atrazine y pendimethalin no fue influenciada por la adición de vinaza, pero la aplicación de biochar redujo el daño causado por atrazine y pendimethalin en todas las especies evaluadas. Un experimento de sorción confirmó la alta afinidad del biochar por atrazine y pendimethalin. Análisis de regresión lineal mostraron que las pendientes de las curvas de adsorción de atrazine y pendimethalin fueron 16 y 4 veces mayores en suelo con biochar que en suelo solo. Bajo condiciones de campo, el biochar a 0.5 kg m−2 redujo el control de malezas de atrazine y pendimethalin en 75% y 60%, respectivamente. Estos resultados sugirieron que el uso de biochar como enmienda de suelo en sistemas de cultivos podría disminuir la eficacia de herbicidas PRE. Por esto, prácticas de mitigación tales como el uso de mayores dosis o una mayor dependencia en herbicidas POST y labranza podrían ser necesarios para asegurar un control adecuado de malezas.

Management of Perennial Warm-Season Bioenergy Grasses. II. Seasonal Differences in Elephantgrass and Energycane Morphological Characteristics Affect Responses to Harvest Frequency and Timing

Elephantgrass (Pennisetum purpureum Schum.) and energycane (Saccharum spp. interspecific hybrid) are perennial C4 grasses with potential for use as bioenergy feedstocks. Their biomass production has been quantified, but differences in plant morphology and the relationship of morphology with biomass harvested and plant persistence are not well understood. The objective was to quantify monthly changes in morphological characteristics of elephantgrass (cv. Merkeron and breeding line UF1) and energycane (cv. L 79-1002) and relate these changes to biomass accumulation and plant responses to defoliation. All were evaluated monthly during full-season growth or when defoliated once in mid-season. Merkeron and UF1 elephantgrass generally showed similar morphological characteristics. Relative to energycane, elephantgrass had fewer tillers early in the growing season, less seasonal variation in tiller number, greater tiller mass and maximum leaf area index (LAI), and earlier spring development of LAI. Energycane showed slower leaf area development in spring, lower maximum LAI, and shorter period of increasing tiller mass and canopy height during the growing season relative to UF1. Elephantgrass had greater incidence of lodging than energycane when exposed to high wind, likely due to greater elephantgrass tiller mass. Morphological characteristics of tall-growing bioenergy grasses help to explain differences among them in biomass production and plant persistence responses to defoliation.

Vinasse and Biochar Effects on Germination and Growth of Palmer Amaranth (Amaranthus palmeri), Sicklepod (Senna obtusifolia), and Southern Crabgrass (Digitaria ciliaris)

Abstract Vinasse and biochar are by-products of biofuel production that can be used as sources of nutrients to crops or soil amendments to improve soil quality. Despite the recent interest in biochar and vinasse effects on soil properties, little is known about their effect on weed communities. We hypothesized that the addition of biochar and vinasse to the soil could affect weed seed germination and growth, and that different weed species would show different responses to these soil amendments. Therefore, the objectives of this study were to determine the effects of vinasse and biochar on the germination and growth of Palmer amaranth, sicklepod, and southern crabgrass. The study was conducted under laboratory and growth chamber conditions. Treatments consisted of four levels of vinasse (0, 10, 20, and 40 L m−2) and biochar (0, 0.5, 2.5, and 12.5 kg m−2) applied to a sandy loam soil. Biochar at 0.5 and 2.5 kg m−2 increased germination of Palmer amaranth but had no effect on sicklepod and southern crabgrass. Vinasse reduced germination of all species. However, sicklepod germination was less affected by vinasse at 10 and 20 L m−2 than the other two species. Vinasse at 40 L m−2 decreased Palmer amaranth, southern crabgrass and sicklepod germination 57, 26 and 87%, respectively. Biochar had no consistent effect on the vegetative growth of the species studied. Vinasse at 10 L m−2 stimulated growth of sicklepod and southern crabgrass compared to the nontreated control. Our results suggested that vinasse used as a soil amendment could affect weed community structure by decreasing germination of susceptible species, but plants and weed species that can get established in vinasse amended soils might show higher growth rates. Nomenclature: Biochar; vinasse; Palmer amaranth, Amaranthus palmeri S. Wats, AMAPA; sicklepod, Senna obtusifolia (L.) H.S. Irwin & Barneby CASOB; southern crabgrass, Digitaria ciliaris (Retz) Koel DIGSP. Resumen La vinaza y el biochar son subproductos de la producción de biocombustibles que pueden ser utilizados como fuentes de nutrientes para cultivos o como enmiendas para mejorar la calidad del suelo. A pesar del reciente interés en los efectos del biochar y la vinaza sobre las propiedades del suelo, es poca la información disponible sobre su efecto en las comunidades de malezas. Planteamos la hipótesis de que la adición de biochar o vinaza al suelo podría afectar la germinación y el crecimiento de malezas, y que la respuesta a estas enmiendas puede ser distinta dependiendo de la especie de malezas. Por lo tanto, los objetivos de este estudio fueron determinar los efectos de la vinaza y el biochar en la germinación y crecimiento de Amaranthus palmeri, Senna obtusifolia y Digitaria ciliaris. El estudio fue realizado en condiciones de laboratorio y cámara de crecimiento. Los tratamientos consistieron en cuatro niveles de vinaza (0, 10, 20, and 40 L m−2) y biochar (0, 0.5, 2.5, and 12.5 kg m−2) aplicados a un suelo franco arenoso. Biochar a 0.5 y 2.5 kg m−2 incrementó la germinación de A. palmeri, pero no tuvo efecto en S. obtusifolia ni en D. ciliaris. La vinaza redujo la germinación de todas las especies. Sin embargo, la germinación de S. obtusifolia fue menos afectada que las otras dos especies cuando se usó vinaza a 10 y 20 L m−2. Vinaza a 40 L m−2 redujo la germinación de A. palmeri, D. ciliaris and S. obtusifolia en 57, 26 and 87%, respectivamente. Biochar no tuvo un efecto consistente sobre el crecimiento vegetativo de las especies estudiadas. Vinaza a 10 L m−2 estimuló el crecimiento de S. obtusifolia and D. ciliaris al comparase con el testigo no-tratado. Nuestros resultados sugieren que el uso de vinaza como enmienda para el suelo podría afectar la estructura de las comunidades de malezas al disminuir la germinación de especies susceptibles de malezas. Sin embargo, las malezas que logren establecerse en suelo enmendado con vinaza podrían mostrar mayores tasas de crecimiento.

Tissue chemistry and morphology affect root decomposition of perennial bioenergy grasses on sandy soil in a sub‐tropical environment

Second‐generation biofuels and bio‐based products derived from lignocellulosic biomass are likely to replace current fuels derived from simple sugars and starch because of greater yield potential and less competition with food production. Besides the high aboveground biomass production, these bioenergy grasses also exhibit extensive root systems. The decomposition of root biomass greatly influences nutrient cycling and microbial activity and subsequent accumulation of carbon (C) in the soil. The objective of this research was thus to characterize root morphological and chemical differences in six perennial grass species in order to better understand root decomposition and belowground C cycling of these bioenergy cropping systems. Giant reed (Arundo donax), elephantgrass (Pennisetum purpureum), energycane (Saccharum spp.), sugarcane (Saccharum spp.), sweetcane (Saccharum arundinaceum), and giant miscanthus (Miscanthus × giganteus) were established in Fall 2008 in research plots near Gainesville, Florida. Root decomposition rates were measured in situ from root decomposition bags over 12 months along with initial and final root tissue composition. Root potential decomposition rate constant (K) was higher in elephantgrass (3.64 g kg−1 day−1) and sweetcane (2.77 g kg−1 day−1) than in sugarcane (1.62 g kg−1 day−1) and energycane (1.48 g kg−1 day−1). Notably, K was positively related to initial root tissue total C (Total C), total fiber glucose (TFG), total fiber xylose (TFX), and total fiber carbohydrate (TFC) concentrations, but negatively related to total fiber arabinose (TFA) and lignin (TL) concentrations and specific root volume (SRV). Among the six species, elephantgrass exhibited root traits most favorable for fast decomposition: high TFG, high TFX, high TFC, high specific root length (SRL), and a low SRV, whereas giant reed, sugarcane, and energycane exhibited slow decomposition rates and the corresponding root traits. Thus, despite similar aboveground biomass yields in many cases, these species are likely to differentially affect soil C accumulation.