Sweet Sorghum Juice and Bagasse as Feedstocks for the Production of Optically Pure Lactic Acid by Native and Engineered <Emphasis Type="Italic">Bacillus coagulans</Emphasis> Strains

Sweet sorghum is a bioenergy crop that produces large amounts of soluble sugars in its stems (3–7 Mg ha^?1) and generates significant amounts of bagasse (15–20 Mg ha^?1) as a lignocellulosic feedstock. These sugars can be fermented not only to biofuels but also to bio-based chemicals. The market potential of the latter may be higher given the current prices of petroleum and natural gas. The yield and rate of production of optically pure d-(?)- and l-(+)-lactic acid as precursors for the biodegradable plastic polylactide was optimized for two thermotolerant Bacillus coagulans strains. Strain 36D1 fermented the sugars in unsterilized sweet sorghum juice at 50 °C to l-(+)-lactic acid (～150 g L^?1; productivity, 7.2 g L^?1 h^?1). B. coagulans strain QZ19-2 was used to ferment sorghum juice to d-(?)-lactic acid (～125 g L^?1; productivity, 5 g L^?1 h^?1). Carbohydrates in the sorghum bagasse were also fermented after pretreatment with 0.5 % phosphoric acid at 190 °C for 5 min. Simultaneous saccharification and co-fermentation of all the sugars (SScF) by B. coagulans resulted in a conversion of 80 % of available carbohydrates to optically pure lactic acid depending on the B. coagulans strain used as the microbial biocatalyst. Liquefaction of pretreated bagasse with cellulases before SScF (L + SScF) increased the productivity of lactic acid. These results show that B. coagulans is an effective biocatalyst for fermentation of all the sugars present in sweet sorghum juice and bagasse to optically pure lactic acid at high titer and productivity as feedstock for bio-based plastics.     

Juice,sugar, and bagasse response of sweet sorghum (Sorghum bicolor (L.) Moench cv. M81E) to N fertilization and soil type

The objective of this research was to determine the optimum nitrogen fertilizer rate for producing sweet sorghum (a promising biofuel crop) juice, sugar, and bagasse on silt loam, sandy loam, and clay soils in Missouri. Seven nitrogen fertilization rates were applied, ranging from 0 to 134 kg N ha^?1. Regardless of the soil and year, the juice content of sweet sorghum stalk averaged 68.8% by weight. The juice yield ranged from 15.2 to 71.1 m³ ha^?1. Soil and N rate significantly impacted the juice yield (P < 0.0001). The pH and the density of the juice were not affected by the soil or N. The sugar content (Brix) of the juice varied between 10.7% and 18.9%. N fertilization improved the sugar content of the juice. A negative correlation existed between the sugar concentration and the juice yield. In general, the lowest sugar content was found in the clay soil and the impact of the N fertilization on juice sugar content was most pronounced in that soil. The juice sugar yield ranged between 2 and 9.9 Mg ha^?1, with significant differences found between years, N rates, and soils. N fertilization always increased the sugar yield in the clay soil, whereas in loam soil, a significant sugar response was recorded when the sweet sorghum was planted after corn. The average juice water content was 84% by weight. The dry bagasse yield fluctuated between 3.2 and 13.8 Mg ha^?1 with significant difference found with N rate, soil, and year. When sweet sorghum was grown after soybean or cotton, its N requirement was less than after a corn crop was grown the previous year. In general, a minimum of 67 kg N ha^?1 was required to optimize juice, sugar, and bagasse yield in sweet sorghum.     

Bagasse Silage from Sweet Pearl Millet and Sweet Sorghum as Influenced by Harvest Dates and Delays between Biomass Chopping and Pressing

Bagasse remaining after extracting the juice from crop biomass for ethanol production could be preserved as silage and used in animal feedstock, but the nutritive and conservation attributes of bagasse silage from sweet sorghum (Sorghum bicolor (L.) Moench) and sweet pearl millet (Pennisetum glaucum (L.) R.Br) are not well known. We evaluated the nutritive and conservation attributes of silages made with the bagasse of two species (sweet pearl millet and sweet sorghum) harvested on two dates (August and September) at two sites in Québec (Canada) and ensiled after four delays between biomass chopping and pressing (0.5, 2, 4, and 6 h). Bagasse silages made in laboratory silos were considered well preserved (pH?≤?4.0, NH₃-N?<?100 g kg^?1 total N, lactate?>?30 g kg^?1 DM, no propionic and butyric acids) regardless of species, harvest date, or delay between biomass chopping and pressing. Sweet pearl millet and sweet sorghum bagasse silages had similar total N concentration, in vitro true digestibility of dry matter (IVTD), and in vitro neutral detergent fiber digestibility (NDFD). Bagasse silage made from biomass harvested in August rather than in September had a 4 % greater concentration of total N, a 4 % greater IVTD, and a 8 % greater NDFD. The delay between biomass chopping and pressing did not affect the nutritive and conservation attributes of silages. Juice extraction from the biomass of sweet pearl millet and sweet sorghum did not impair attributes of good silage fermentation but it reduced its nutritive value.     

Assessment of Stalk Properties to Predict Juice Yield in Sorghum

Sweet sorghum is an outstanding feedstock choice for bioethanol production, but the gap between theoretical and commercial ethanol yields must be reduced to improve economic viability. Extractable juice yield is a primary limiting factor for higher ethanol yield, but current phenotyping techniques to measure juice yield in sorghum can be laborious. Therefore, alternative approaches to measuring juice yield during selection are needed. The objectives of this study were to investigate the relationship between stalk-related traits and juice yield and to assess the ability to predict juice yield using agronomic traits and stalk properties across and within a diverse set of sorghum ideotypes (photoinsensitive, photosensitive, biomass, grain, and sweet types). Stalk weight, stalk volume, stalk diameter, and plant height had significantly strong associations with juice yield, which were consistent across different sorghum ideotypes. The direct and indirect effects of multiple predictive traits on juice yield varied greatly with the distinct sorghum subsets. However, equation modeling demonstrated that juice yield is satisfactorily predicted by jointly assessing stalk weight and stalk moisture. Moreover, alternative prediction models involving distinct combinations of agronomic and stalk-related traits had similarly good prediction accuracy. Altogether, this suggests that several prediction models can be used to accelerate phenotyping for juice yield, which will improve the selection process. Overall, the results indicate that increasing sorghum juice yield via indirect selection is possible, but the choice of prediction model depends on the ideotypes and resources available in a breeding program.     

Assessment of Sugarcane Billet Harvester on Recovery of Sweet Sorghum Biomass for Ethanol Production

Sweet sorghum (Sorghum bicolor L. Moench) is a promising bioenergy crop for the production of ethanol and bio-based products. Sugarcane billet harvesters can be used to harvest sweet sorghum. Multiple extractor fan speed settings of these harvesters allow for separating the extraneous matter in the feedstock, which has been associated with increased milling throughput and better juice quality at the processing facility. This removal is not completely selective, and some stalk material is also lost. These losses can be higher for sweet sorghum than sugarcane due its lower weight. This paper presents an assessment of how the speed of the primary extractor fan of a sugarcane billet combine used for harvesting sweet sorghum affects the biomass yield, biomass losses, and quality at delivery for the production of ethanol from extracted juice and fiber. Three primary extractor fan speeds (0, 800, and 1100 rpm) were evaluated. Higher fan speeds decreased fresh biomass yields by up to 28.3 Mg ha^?1. Juice quality was not significantly different among treatments. Ethanol yield calculated from sweet sorghum harvested at 0 rpm was 6075 L ha^?1. This value decreased by about half for material harvested at 1100 rpm due to the differences in biomass yield.     

Evaluation of Public Sweet Sorghum A-Lines for Use in Hybrid Production

A fundamental need for commercialization of sweet sorghum [Sorghum bicolor (L.) Moench] as a bioenergy crop is an adequate seed supply, which will require development of hybrid varieties using dwarf seed-parent lines. A set of six public sweet sorghum A-lines (Dwarf Kansas Sourless, KS9, N36, N38, N39, and N4692) were crossed with a set of six public sweet sorghum cultivars (Brawley, Kansas Collier, Dale, Sugar Drip, Waconia, and Wray). Grain, fiber, and sugar yields were determined, and conversion formulas were applied to estimate ethanol yields. Hybrids were grown in fields at Ithaca, NE, USA, in 1983–1984 fertilized with 112 kg ha^?1 N. In terms of yield components and overall ethanol yields, one A-line, N38, was inferior. Average total ethanol yields from hybrids made on the other A-lines were not significantly different, suggesting that any of those five A-lines could be useful seed-parents. With the exception of grain yield, cultivars used as pollen parents were among the highest-performing entries for all traits. For all traits directly contributing to total ethanol yield (grain yield, juice yield, % soluble solids, sugar yield, fiber yield), hybrids were also among the highest-performing entries. Results of this study demonstrate that hybrid sweet sorghum with performance criteria equivalent to existing sweet sorghum cultivars can be produced on the sweet sorghum seed-parent lines A-Dwarf Kansas Sourless, A-KS9, A-N36, A-N39, and A-N4692. Identification of specific seed-parent × pollen parent lines with characteristics best suited for particular growing regions and end-user needs will be critical for commercial hybrid development.     

Morphological and molecular characterization of sweet,grain and forage sorghum (Sorghum bicolor L.) genotypes grown under temperate climatic conditions

Abstract

In the present study, we used 12 genotypes of sorghum originated from different countries (five sweet, four grain and three forage). These different genotypes and types of sorghum were evaluated for the agro-morphological traits that are associated with the estimated sugar and bioethanol yield to estimate their phenotypic diversity. Analysis of variance showed significant differences between different types of sorghum for all the evaluated traits. Sweet sorghum genotypes, however, showed better performance with respect to all studied traits than the other genotypes. A positive significant correlation was observed between plant height, leaf number, leaf area, biomass yield, cane and bagasse yields, and the predicted bioethanol yield. Both, cluster and principal component analysis were performed to group the genotypes according to their agro-morphological and molecular similarity coefficients. For analytical approaches, the Iranian grain and forage genotypes clustered separately from the other genotypes. The clustering patterns obtained from the molecular dominant markers had higher discriminatory power than using morphological characters to separate sweet genotypes from the forage and grain sorghum ones. The results clearly indicated that sweet sorghum can be grown in Germany and maintains its superiority in biomass production and sugar yield over grain and forage sorghum types.     

Techno-Economic Evaluation of Cellulosic Ethanol Production Based on Pilot Biorefinery Data: a Case Study of Sweet Sorghum Bagasse Processed via L+SScF

Replacing fossil fuels with renewable fuels derived from lignocellulosic biomass can contribute to the mitigation of global warming and the economic development of rural communities. This will require lignocellulosic biofuels to become price competitive with fossil fuels. Techno-economic analyses can provide insights into which parts of the biofuel production process need to be optimized to reduce cost or energy use. We used data obtained from a pilot biorefinery to model a commercial-scale biorefinery that processes lignocellulosic biomass to ethanol, with a focus on the minimum ethanol selling price (MESP). The process utilizes a phosphoric acid-catalyzed pre-treatment of sweet sorghum bagasse followed by liquefaction and simultaneous saccharification and co-fermentation (L+SScF) of hexose and pentose sugars by an engineered Escherichia coli strain. After validating a techno-economic model developed with the SuperPro Designer software for the conversion of sugarcane bagasse to ethanol by comparing it to a published Aspen Plus model, six different scenarios were modeled for sweet sorghum bagasse Under the most optimistic scenario, the ethanol can be produced at a cost close to the energy-equivalent price of gasoline. Aside from an increase in the price of gasoline, the gap between ethanol and gasoline prices could also be bridged by either a decrease in the cost of cellulolytic enzymes or development of value-added products from lignin.     

A holistic high-throughput screening framework for biofuel feedstock assessment that characterises variations in soluble sugars and cell wall composition in <Emphasis Type="Italic">Sorghum bicolor</Emphasis>

Background

A major hindrance to the development of high yielding biofuel feedstocks is the ability to rapidly assess large populations for fermentable sugar yields. Whilst recent advances have outlined methods for the rapid assessment of biomass saccharification efficiency, none take into account the total biomass, or the soluble sugar fraction of the plant. Here we present a holistic high-throughput methodology for assessing sweet Sorghum bicolor feedstocks at 10 days post-anthesis for total fermentable sugar yields including stalk biomass, soluble sugar concentrations, and cell wall saccharification efficiency.

Results

A mathematical method for assessing whole S. bicolor stalks using the fourth internode from the base of the plant proved to be an effective high-throughput strategy for assessing stalk biomass, soluble sugar concentrations, and cell wall composition and allowed calculation of total stalk fermentable sugars. A high-throughput method for measuring soluble sucrose, glucose, and fructose using partial least squares (PLS) modelling of juice Fourier transform infrared (FTIR) spectra was developed. The PLS prediction was shown to be highly accurate with each sugar attaining a coefficient of determination (R ² ) of 0.99 with a root mean squared error of prediction (RMSEP) of 11.93, 5.52, and 3.23 mM for sucrose, glucose, and fructose, respectively, which constitutes an error of <4% in each case. The sugar PLS model correlated well with gas chromatography–mass spectrometry (GC-MS) and brix measures. Similarly, a high-throughput method for predicting enzymatic cell wall digestibility using PLS modelling of FTIR spectra obtained from S. bicolor bagasse was developed. The PLS prediction was shown to be accurate with an R ² of 0.94 and RMSEP of 0.64 μg.mgDW^-1.h^-1.

Conclusions

This methodology has been demonstrated as an efficient and effective way to screen large biofuel feedstock populations for biomass, soluble sugar concentrations, and cell wall digestibility simultaneously allowing a total fermentable yield calculation. It unifies and simplifies previous screening methodologies to produce a holistic assessment of biofuel feedstock potential.

     

Genetic mapping of QTLs for sugar-related traits in a RIL population of Sorghum bicolor L. Moench

The productivity of sorghum is mainly determined by quantitative traits such as grain yield and stem sugar-related characteristics. Substantial crop improvement has been achieved by breeding in the last decades. Today, genetic mapping and characterization of quantitative trait loci (QTLs) is considered a valuable tool for trait enhancement. We have investigated QTL associated with the sugar components (Brix, glucose, sucrose, and total sugar content) and sugar-related agronomic traits (flowering date, plant height, stem diameter, tiller number per plant, fresh panicle weight, and estimated juice weight) in four different environments (two locations) using a population of 188 recombinant inbred lines (RILs) from a cross between grain (M71) and sweet sorghum (SS79). A genetic map with 157 AFLP, SSR, and EST-SSR markers was constructed, and several QTLs were detected using composite interval mapping (CIM). Further, additive × additive interaction and QTL × environmental interaction were estimated. CIM identified more than five additive QTLs in most traits explaining a range of 6.0–26.1% of the phenotypic variation. A total of 24 digenic epistatic locus pairs were identified in seven traits, supporting the hypothesis that QTL analysis without considering epistasis can result in biased estimates. QTLs showing multiple effects were identified, where the major QTL on SBI-06 was significantly associated with most of the traits, i.e., flowering date, plant height, Brix, sucrose, and sugar content. Four out of ten traits studied showed a significant QTL × environmental interaction. Our results are an important step toward marker-assisted selection for sugar-related traits and biofuel yield in sorghum.     

Evaluation of <Emphasis Type="Italic">ZmCCT</Emphasis> haplotypes for genetic improvement of maize hybrids

Key message

The elite ZmCCT haplotypes which have no transposable element in the promoter could enhance maize resistance to Gibberella stalk rot and improve yield-related traits, while having no or mild impact on flowering time. Therefore, they are expected to have great value in future maize breeding programs.

Abstract

A CCT domain-containing gene, ZmCCT, is involved in both photoperiod response and stalk rot resistance in maize. At least 15 haplotypes are present at the ZmCCT locus in maize germplasm, whereas only three of them are found in Chinese commercial maize hybrids. Here, we evaluated ZmCCT haplotypes for their potential application in corn breeding. Nine resistant ZmCCT haplotypes that have no CACTA-like transposable element in the promoter were introduced into seven elite maize inbred lines by marker-assisted backcrossing. The resultant 63 converted lines had 0.7-5.1 Mb of resistant ZmCCT donor segments with over 90% recovery rates. All converted lines tested exhibited enhanced resistance to maize stalk rot but varied in photoperiod sensitivity. There was a close correlation between the hybrids and their parental lines with respect to both resistance performance and photoperiod sensitivity. Furthermore, in a given hybrid A5302/83B28, resistant ZmCCT haplotype could largely improve yield-related traits, such as ear length and 100-kernel weight, resulting in enhanced grain yield. Of nine resistant ZmCCT haplotypes, haplotype H5 exhibited excellent performance for both flowering time and stalk rot resistance and is thus expected to have potential value in future maize breeding programs.

     

Heat susceptibility of grain filling in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) linked with rapid chlorophyll loss during a 3-day heat treatment

Brief heat events (1–3 days, >30 °C) commonly reduce wheat (Triticum aestivum L.) grain size and consequently yield. To identify mechanisms of tolerance to such short heat events, 36 wheat genotypes were treated under day/night temperatures of 37 °C/27 °C for 3-days in a growth chamber, at 10 days after anthesis, and a range of developmental, chlorophyll and yield-related traits monitored. The degree of flag leaf chlorophyll loss during the treatment was the variable that showed the highest correlation to grain weight loss (r = 0.63; p < 0.001), identifying chlorophyll stability during this brief period as a potential determinant or indicator of grain weight stability under heat. Variables summarizing the combined during- and post-heat chlorophyll losses showed similar or lower correlations with heat tolerance of grain filling, despite the fact that genotypes varied in their ability to resume normal chlorophyll loss rates after the heat treatment. Additionally, heat tolerance of grain size showed no correlation with grain filling duration or traits relating to utilization of stem carbon reserves under heat stress. Measurement of chlorophyll loss over a forecasted heat wave was thereby identified as a potential basis for developing tools to help breeders select heat tolerant genotypes.     

Identifying Morphological and Mechanical Traits Associated with Stem Lodging in Bioenergy Sorghum (<Emphasis Type="Italic">Sorghum bicolor</Emphasis>)

Stem lodging in Sorghum is a major agronomic problem that has far-reaching economic consequences. More rapid and reliable advances in stem lodging resistance could be achieved through development of selective breeding tools that are not dependent on post hoc data or dependent on abiotic or biotic environmental factors. Our objective was to use sorghum to examine how mechanical stability is achieved and lost, and to provide insights into the development of a rapid and reliable phenotyping approach. The biomechanical properties of the stems of six bioenergy sorghum genotypes were investigated using a three-point bending test protocol. Important morphometric data were also collected, and previously collected lodging scores were used to associate with morphological and mechanical traits. Nodes were two to three-folds stronger, stiffer, and more rigid than internodes. In general, internodes were numerically weakest and more rigid between internodes 3 and 6, corresponding to the area where higher stem lodging is observed. Internode strength was negatively correlated with diameter (r = ?0.77, P < 0.05) and volume (r = 0.96, P < 0.01), while stem lodging was positively correlated with flexural rigidity (r = 0.85, P < 0.05) and volume (r = 0.78, P < 0.05). The analysis revealed key functional traits that influence the mode and location of stem lodging. Moreover, these results indicate the potential of these methods as a selective breeding tool for indirect selection of stem lodging resistance in bioenergy sorghum.     

Genetic diversity of some Iranian sweet cherry (<Emphasis Type="Italic">Prunus avium</Emphasis>) cultivars using microsatellite markers and morphological traits

The aim of this study was to characterize 23 important Iranian sweet cherry (Prunus avium) cultivars collected from different provinces of Iran and 1 foreign cultivar, which was used as control, considered for breeding programs by using 21 microsatellite markers and 27 morphological traits. In sweet cherry (Prunus avium) accessions, leaf, fruit, and stone morphological characters were evaluated during two consecutive years. The study revealed a high variability in the set of evaluated sweet cherry accessions. The majority of important correlations were determined among variables representing fruit and leaf size and variables related to color. Cluster analysis distinguished sweet cherry accessions into two distinct groups. Principal component analysis (PCA) of qualitative and quantitative morphological parameters explained over 86.59% of total variability in the first seven axes. In PCA, leaf traits such as leaf length and width, and fruit traits such as length, width, and weight, and fruit flesh and juice color were predominant in the first two components, indicating that they were useful for the assessment of sweet cherry germplasm characterization. Out of 21 SSR markers, 16 were polymorphic, producing 177 alleles that varied from 4 to 16 alleles (9.35 on average) with a mean heterozygosity value of 0.82 that produced successful amplifications and revealed DNA polymorphisms. Allele size varied from 95 to 290 bp. Cluster analyses showed that the studied sweet cherry genotypes were classified into five main groups based mainly on their species characteristics and SSR data. In general, our results did not show a clear structuring of genetic variability within the Iranian diffusion area of sweet cherry, so it was not possible to draw any indications on regions of provenance delimitation. The results of this study contribute to a better understanding of sweet cherry genetic variations in Iran, thus making for more efficient programs aimed at preserving biodiversity and more rational planning of the management of reproductive material.     

Evaluation and Modeling of Bioethanol Yield Efficiency from Sweet Sorghum Juice

One of the challenges with using sweet sorghum as an energy crop is that although fermentation of the juice to ethanol does not require enzymes, the juice can easily spoil. One strategy to avoid spoilage is to harvest the juice in the field, place it into a tanker for transport, and add the yeast immediately to initiate the fermentation process to begin during transport. Hence, it is also important to understand how the fermentation process is influenced by pH, temperature, and dissolved oxygen, since these parameters would not be “controlled” during transport. A full factorial design was applied to examine and optimize yield efficiency of ethanol production for the fermentation of sweet sorghum juice. Bioethanol yield efficiency was modeled using a linear equation. Under optimal pH (5.5), temperature (28 °C), and dissolved oxygen (0%) conditions, a maximum theoretical yield efficiency of 0.75 was achieved for bioethanol produced from M81E variety of sweet sorghum.     

Impact of Storage Time on the Juice and Sugars Extracted from Chopped and Whole Stalk Sweet Pearl Millet and Sweet Sorghum Biomass

Since they have a high concentrations of fermentable sugars, sweet pearl millet and sweet sorghum are two interesting crops for bioethanol production. However, if the juice is not extracted from the biomass immediately after harvest, the biomass has to be transported and stored for further juice extraction. This delay could affect the amount of juice extracted and its sugar concentration. This paper presents the results of 3 years of experiments where different storage modes (chopped and whole stalks) and various storage time (0 to 14 days) were applied on two different crop species (sweet pearl millet and sweet sorghum). Storing sweet pearl millet as whole stalks for 2 weeks resulted in a water-soluble carbohydrate (WSC) concentration decrease of 52 %, while no significant decrease of the WSC concentration was observed for sweet sorghum. Whole stalks storage is much more efficient than storing the biomass chopped to avoid a rapid sugar loss. However, more juice can be extracted from stored chopped biomass than from stored whole stalks biomass. If the juice cannot be extracted quickly after the harvest, the biomass can be stored as whole stalks to avoid rapid sugar deterioration, especially for sweet sorghum.     

Genetic transformation and evaluation of two sweet sorghum genotypes for resistance to spotted stemborer, <Emphasis Type="Italic">Chilo partellus</Emphasis> (Swinhoe)

Sweet sorghum is a climate smart crop with multiple uses. The crop is susceptible to attack by the spotted stemborer, Chilo partellus (Swinhoe). This causes deadheart formation, leading to lodging of plants and consequent high economic losses. Lack of stable sources of resistance make any genetic enhancement through breeding difficult. We report a study to build up host plant resistance using transgenic technology by introducing two different classes of Bt genes (cry1Aa and cry1B) into two elite sweet sorghum genotypes of India (SSV84 and RSSV9). We devised tissue culture methods to suit the genotypes of our interest, SSV84 and RSSV9, and employed two methods of genetic transformation: the particle bombardment and in planta method of Agrobacterium. Modification of in vitro culture methods involved subculture every 3 days in the initial stages of culture and the use of precultured embryos as target tissues. For the in planta method, a floral dip for 1 h in Agrobacterium suspension supplemented with l-cysteine and Tween-20 was used. Sixteen transgenic events were generated; inheritance, integration and stable expression of the transgenes till the T₄ generation were confirmed. The amount of Bt Cry1Aa protein at 25–30 days of growth ranged from 24.8 to 72.8 ng/g of fresh leaf tissue. We recorded 78.4 % larval mortality, reduced leaf damage (3.0 out of 9.0) and reduced feeding (41.0 %) over the controls in insect feed assays. Stable inheritance and expression in the in planta-derived transgenics are presented.     

Composition of sugar cane,energy cane,and sweet sorghum suitable for ethanol production at Louisiana sugar mills

A challenge facing the biofuel industry is to develop an economically viable and sustainable biorefinery. The existing potential biorefineries in Louisiana, raw sugar mills, operate only 3 months of the year. For year-round operation, they must adopt other feedstocks, besides sugar cane, as supplemental feedstocks. Energy cane and sweet sorghum have different harvest times, but can be processed for bio-ethanol using the same equipment. Juice of energy cane contains 9.8% fermentable sugars and that of sweet sorghum, 11.8%. Chemical composition of sugar cane bagasse was determined to be 42% cellulose, 25% hemicellulose, and 20% lignin, and that of energy cane was 43% cellulose, 24% hemicellulose, and 22% lignin. Sweet sorghum was 45% cellulose, 27% hemicellulose, and 21% lignin. Theoretical ethanol yields would be 3,609 kg per ha from sugar cane, 12,938 kg per ha from energy cane, and 5,804 kg per ha from sweet sorghum.     

Fumaric Acid Production by <Emphasis Type="Italic">Rhizopus oryzae</Emphasis> ATCC® 20344™ from Lignocellulosic Syrup

Powdered activated carbon-treated lignocellulosic syrup prepared from energy cane bagasse was evaluated as a potential feedstock in the production of fumaric acid by Rhizopus oryzae ATCC® 20344?. Energy cane bagasse was pretreated with dilute ammonia and enzymatically hydrolyzed with commercially available enzymes, Cellic® CTec2 and HTec2. The collected hydrolysate samples were subjected to powdered activated carbon adsorption for the removal of non-sugar compounds (i.e., organic acids, furaldehydes, total phenolic compounds) and concentrated to a final 65°Bx syrup (mostly xylose and glucose sugars). The use of lignocellulosic syrup, the effect of nitrogen source, medium additives, and initial pH in the seed culture medium on fungal morphology were investigated. The carbon to nitrogen (C/N) ratio in the acid production medium was also optimized for maximum yields in fumaric acid production. Optimum seed culture medium conditions (2.0 g/L urea, 3.0 pH) produced the desired compact, smooth, and uniform fungal pellets. Optimum acid production medium conditions (400 C/N ratio, 0.2 g/L urea) resulted in a fumaric acid production of 34.20 g/L, with a yield of 0.43 g/g and a productivity of 0.24 g/L/h. These results were comparable to those observed with the control medium (pure glucose and xylose). The present study demonstrated that lignocellulosic syrup processed from dilute ammonia pretreated energy cane bagasse has potential as a renewable carbon source for fumaric acid fermentation by Rhizopus oryzae ATCC® 20344?.     

Overexpression of the Bx7 high molecular weight glutenin subunit on the <Emphasis Type="Italic">Glu</Emphasis>-<Emphasis Type="Italic">B1</Emphasis> locus in a Korean wheat landrace

The Glu-B1al (Bx7^OE + By8) allele is important for bread-making quality. The allele was found in a Korean wheat landrace using specific DNA markers. Molecular analyses were conducted to identify the overexpressed Bx7 (Bx7^OE) subunit of the allele. The Korean wheat landrace (accession ID: IT166460) showed a similar protein expression level of Bx7 subunit, i.e., overexpression of Bx7 subunit towards cv. Glenlea, Canadian Western Red Spring wheat, which harbors Bx7^OE subunit of Glu-B1al as detected on SDS–PAGE (sodium dodecyl sulfate poly-acrylamide gel electrophoresis). In addition, 2-DE (two-dimensional electrophoresis) analysis revealed similar protein expression patterns of the Bx7 subunit regions of IT166460 and Glenlea. The proportion of Bx7 to total HMW-GSs (high molecular weight glutenin subunits) in IT166460 (56.17 ± 0.22%) was higher than that of Chinese Spring (34.75 ± 1.03%) and even that of Glenlea (46.25 ± 1.76%) as assessed by RP-HPLC (reverse-phase high-performance liquid chromatography). Overexpression of Bx7 subunit was caused by gene duplication and indels of the promoter region of the Bx7 gene. IT166460 attained the 43 bp indel of the promoter region, as did Glenlea, i.e., the amplicon size of IT166460 was the same as that of Glenlea. In addition, the nucleotides present in the duplicated gene in IT166460 were the same as those in Glenlea. Bx7^OE subunit is critical for dough strength. However, most wheat accessions harboring the subunit are distributed in America. Furthermore, most Korean wheats have little genetic variation in glutenin composition and are associated with inferior bread quality. Hence, IT166460 could be used to improve bread-making quality in the Korean wheat breeding program.