Perspective of the Sugarcane Industry in Brazil

The sugarcane industry in Brazil is experiencing a rapid shift towards creating the grounds for a green and sustainable biorefinary industry. After 30 years of ProAlcool, the federal government program that boosted Brazil’s sugarcane industry by creating a mandate to blend ethanol with gasoline, flex fuel engines now dominate Brazil’s automobile industry. Currently, bioethanol replaces around 30% of the gasoline consumed in the country and its demand is projected to more that double in the next 10 years. On another front, the sugarcane genomics program created by FAPESP in the late 1990s paved the way for the establishment of innovative biotechnology startup companies that attracted the attention of the largest agro-biotechnology sector companies of the world. Almost all of these companies now have their sugarcane research centers surrounding the city of Campinas, São Paulo. In addition, innovative synthetic biology companies are developing technologies to produce diesel, jet fuel and other high value molecules using sugarcane juice as a carbon source. The sugarcane industry also teamed with petrochemical companies and already established operating plants to produce bioplastics. Innovations have also occurred in the field of co-generation of electricity from sugarcane bagasse. Currently sugarcane supplies 4% of the electricity needs of the country. Collectively, these innovations suggest that Brazil’s sugarcane industry could supply over 30% of the country energy needs by 2021 and a significant fraction of new bioproducts produced by its nascent biorefinary plants.     

Scientific challenges of bioethanol production in Brazil

Bioethanol (fuel alcohol) has been produced by industrial alcoholic fermentation processes in Brazil since the beginning of the twentieth century. Currently, 432 mills and distilleries crush about 625 million tons of sugarcane per crop, producing about 27 billion liters of ethanol and 38.7 million tons of sugar. The production of bioethanol from sugarcane represents a major large-scale technology capable of producing biofuel efficiently and economically, providing viable substitutes to gasoline. The combination of immobilization of CO₂ by sugarcane crops by photosynthesis into biomass together with alcoholic fermentation of this biomass has allowed production of a clean and high-quality liquid fuel that contains 93% of the original energy found in sugar. Over the last 30 years, several innovations have been introduced to Brazilian alcohol distilleries resulting in the improvement of plant efficiency and economic competitiveness. Currently, the main scientific challenges are to develop new technologies for bioethanol production from first and second generation feedstocks that exhibit positive energy balances and appropriately meet environmental sustainability criteria. This review focuses on these aspects and provides special emphasis on the selection of new yeast strains, genetic breeding, and recombinant DNA technology, as applied to bioethanol production processes.     

Sugarcane (Saccharum X officinarum): A Reference Study for the Regulation of Genetically Modified Cultivars in Brazil

Global interest in sugarcane has increased significantly in recent years due to its economic impact on sustainable energy production. Sugarcane breeding and better agronomic practices have contributed to a huge increase in sugarcane yield in the last 30?years. Additional increases in sugarcane yield are expected to result from the use of biotechnology tools in the near future. Genetically modified (GM) sugarcane that incorporates genes to increase resistance to biotic and abiotic stresses could play a major role in achieving this goal. However, to bring GM sugarcane to the market, it is necessary to follow a regulatory process that will evaluate the environmental and health impacts of this crop. The regulatory review process is usually accomplished through a comparison of the biology and composition of the GM cultivar and a non-GM counterpart. This review intends to provide information on non-GM sugarcane biology, genetics, breeding, agronomic management, processing, products and byproducts, as well as the current technologies used to develop GM sugarcane, with the aim of assisting regulators in the decision-making process regarding the commercial release of GM sugarcane cultivars.     

Ethanol production potential of sweet sorghum assessed using forage fiber analysis procedures

Sweet sorghum (Sorghum bicolor (L.) Moench) is widely recognized as a highly promising biomass energy crop with particular potential to complement sugarcane production in diversified cropping systems. Agronomic assessments have led to identification of four cultivars well suited for such sugarcane‐based production systems in southern Louisiana. Sweet sorghum biofuel production systems are currently being developed, and research producing large sample numbers requiring ethanol yield assessment is anticipated. Fiber analysis approaches developed for forage evaluation appear to be useful for screening such large numbers of samples for relative ethanol yield. Chemical composition, forage fiber characteristics, digestibility, and ethanol production of sweet sorghum bagasse from the four cultivars were assessed. Measures of detergent fiber, lignin, and digestibility were highly correlated with ethanol production (P < 0.01). The best linear regression models accounted for about 80% of the variation among cultivars in ethanol production. Bagasse from the cultivar Dale produced more ethanol per gram of material than any of the other cultivars. This superior ethanol production was apparently associated with less lignin in stems of Dale. Forage evaluation measures including detergent fiber analyses, in vitro digestibility, and an in vitro gas production technique successfully identified the cultivar superior in ethanol yield indicating their usefulness for screening sweet sorghum samples for potential ethanol production in research programs generating large sample numbers from evaluations of germ plasm or agronomic treatments. These screening procedures reduce time and expense of alternatives such as hexose sugar assessment for calculating theoretical ethanol yield.     

Ethanol production from lignocellulosic biomass of energy cane

Ethanol produced from lignocellulosic biomass is a renewable alternative to diminishing petroleum based liquid fuels. The release of many new sugarcane varieties by the United States Department of Agriculture to be used as energy crops is a promising feedstock alternative. Energy cane produces large amounts of biomass that can be easily transported, and production does not compete with food supply and prices because energy cane can be grown on marginal land instead of land for food crops. The purpose of this study was to evaluate energy cane for lignocellulosic ethanol production. Energy cane variety L 79-1002 was pretreated with weak sulfuric acid to remove lignin. In this study, 1.4 M sulfuric acid pretreated type II energy cane had a higher ethanol yield after fermentation by Klebsiella oxytoca without enzymatic saccharification than 0.8 M and 1.6 M sulfuric acid pretreated type II energy cane. Pretreated biomass was inoculated with K. oxytoca for cellulose fermentation and Pichia stipitis for hemicellulose fermentation under simultaneous saccahrification and fermentation (SSF) and separate hydrolysis and fermentation (SHF) conditions. For enzymatic saccharification of cellulose, the cellulase and ??-glucanase cocktail significantly increased ethanol production compared to the ethanol production of fermented acid pretreated energy cane without enzymatic saccharification. The results revealed that energy cane variety L 79-1002 produced maximum cellulosic ethanol under SHF (6995 mg/L) and produced 3624 mg/L ethanol from fermentation of hemicellulosic sugars.     

Use of sugarcane molasses “B” as an alternative for ethanol production with wild-type yeast <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> ITV-01 at high sugar concentrations

Molasses “B” is a rich co-product of the sugarcane process. It is obtained from the second step of crystallization and is richer in fermentable sugars (50–65%) than the final molasses, with a lower non-sugar solid content (18–33%); this co-product also contains good vitamin and mineral levels. The use of molasses “B” for ethanol production could be a good option for the sugarcane industry when cane sugar prices diminish in the market. In a complex medium like molasses, osmotolerance is a desirable characteristic for ethanol producing strains. The aim of this work was to evaluate the use of molasses “B” for ethanol production using Saccharomyces cerevisiae ITV-01 (a wild-type yeast isolated from sugarcane molasses) using different initial sugar concentrations (70–291 g L⁻¹), two inoculum sizes and the addition of nutrients such as yeast extract, urea, and ammonium sulphate to the culture medium. The results obtained showed that the strain was able to grow at 291 g L⁻¹ total sugars in molasses “B” medium; the addition of nutrients to the culture medium did not produce a statistically significant difference. This yeast exhibits high osmotolerance in this medium, producing high ethanol yields (0.41 g g⁻¹). The best conditions for ethanol production were 220 g L⁻¹ initial total sugars in molasses “B” medium, pH 5.5, using an inoculum size of 6 × 10⁶ cell mL⁻¹; ethanol production was 85 g L⁻¹, productivity 3.8 g L⁻¹ h⁻¹ with 90% preserved cell viability.     

Study of sugarcane pieces as yeast supports for ethanol production from sugarcane juice and molasses

Due to the environmental concerns and the increasing price of oil, bioethanol was already produced in large amount in Brazil and China from sugarcane juice and molasses. In order to make this process competitive, we have investigated the suitability of immobilized Saccharomyces cerevisiae strain AS2.1190 on sugarcane pieces for production of ethanol. Electron microscopy clearly showed that cell immobilization resulted in firm adsorption of the yeast cells within subsurface cavities, capillary flow through the vessels of the vascular bundle structure, and attachment of the yeast to the surface of the sugarcane pieces. Repeated batch fermentations using sugarcane supported-biocatalyst were successfully carried out for at least ten times without any significant loss in ethanol production from sugarcane juice and molasses. The number of cells attached to the support increased during the fermentation process, and fewer yeast cells leaked into fermentation broth. Ethanol concentrations (about 89.73–77.13 g/l in average value), and ethanol productivities (about 59.53–62.79 g/l d in average value) were high and stable, and residual sugar concentrations were low in all fermentations (0.34–3.60 g/l) with conversions ranging from 97.67–99.80%, showing efficiency (90.11–94.28%) and operational stability of the biocatalyst for ethanol fermentation. The results of this study concerning the use of sugarcane as yeast supports could be promising for industrial fermentations. L. Liang and Y. Zhang have contributed equally to this work.     

Screening of a broad range of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) germplasm for <Emphasis Type="Italic">in vitro</Emphasis> rapid plant regeneration and development of an early prediction system

Rice has emerged as a model monocot for studies in agriculture and biotechnology due to its relatively small genome and a ready accessibility to plant material. Tissue culture is one of the tools required for genetic transformation and some breeding programs, and the selection of high-frequency regenerator types is essential for success in these technologies. Thirty-three rice entries with agricultural and biotechnological characteristics of interest were screened with the aim to identify the best regenerators. Entries that exhibited between 50% and 90% regeneration frequencies include ‘Taipei-309,’ ‘Super Dwarf,’ ‘Norin’ (japonica types), PI 312777, ‘Ali Combo’ (indica types), ‘STG-S,’ and ‘LA3’ (red rice types). One third of the entries tested were at least two times better at regeneration than the often-cited regenerator ‘Nipponbare.’ Those entries showing at least 85% frequency of greening or somatic embryo formation at 15 or 30 d on regeneration medium ultimately produced whole plants after 45 d on regeneration medium at high frequency (at least 40%); those entries not reaching the 85% threshold of greening by Days 15 or 30 exhibited moderate (15–40%) to low (less than 10%) frequency of whole plant regeneration. This greening response suggests the means for an early prediction system for identification of useful rice regenerator lines, which would be beneficial for high-throughput screening of germplasm as well as for decreasing the time and cost of in vitro culture.     

Drivers leading to higher food prices: biofuels are not the main factor

Since 2007, the overall rise in food prices in the USA was twice that of the overall inflation rate, led by inflation for poultry and dairy products in particular. Prominent studies have indicated that the main drivers associated with the rise in food prices in the past 3 yr are the increased energy costs (and the trickle-down impact on farm input costs) and the devaluation of the US dollar. However, currently, the impact of crude oil as one of the primary drivers in food prices has waned significantly, as crude oil prices have fallen dramatically since late 2008. The data reviewed here debunk the popular myth that food producers, particularly farmers growing corn or soybean for biofuel feedstocks, are making huge profits with high commodity prices. Producers continue to face extraordinary risks in their farming operations, and profit margins have narrowed considerably because of the high input prices driven by high energy costs. One of the primary solutions to the food price inflation is to increase the supply of commodity crops in a responsible way that is both respectful and sustainable regarding economic, social, and environmental aspects. At the center of this solution is implementing and developing new technologies to increase crop yields and nutritional values without increasing the amount of fossil-based inputs used in agriculture. Conventional breeding, molecular breeding, genomics, and biotechnology are pivotal technologies for increasing crop yields to meet these supply needs, combined with the impacts of other new technologies and best management practices in agricultural production.     

A spatially explicit whole-system model of the lignocellulosic bioethanol supply chain: an assessment of decentralised processing potential

Background  

Lignocellulosic bioethanol technologies exhibit significant capacity for performance improvement across the supply chain through the development of high-yielding energy crops, integrated pretreatment, hydrolysis and fermentation technologies and the application of dedicated ethanol pipelines. The impact of such developments on cost-optimal plant location, scale and process composition within multiple plant infrastructures is poorly understood. A combined production and logistics model has been developed to investigate cost-optimal system configurations for a range of technological, system scale, biomass supply and ethanol demand distribution scenarios specific to European agricultural land and population densities.     

How long has the ‘hotspot’ been ‘hot’? Past stand-scale structures at Siggaboda nature reserve in southern Sweden

Fossil pollen and plant macrofossils over the last 2000 years are documented from three small forest hollows in Southern Sweden. One of the sites is inside a 5 ha highly prized old growth mixed Fagus sylvatica and Picea abies forest of high biodiversity which has been protected since 1940. The other two hollows are located 400 and 700 m away in an outlying buffer zone established in 1995 which is mainly coniferous plantation forest. The results show that the area has been forested for at least 2000 years, but that forest composition has been under continuous change, most rapid over the last 200 years. The reduction of deciduous tree pollen particularly Quercus, Tilia, Alnus and Corylus, and the immigration of Fagus and Picea can be observed at all three sites. However, the temperate deciduous trees (Quercus or Fagus) have been much more common in the ‘hotspot’ than in the surrounding forests over the last c. 200 years, and significantly more common at least 2000 years before that. Even though the vegetation has been dynamic through time, the lower human intervention in the ‘hotspot’ area compared with the surrounding matrix forests has facilitated the longevity of deciduous trees and the many rare species which are associated with them. The palaeoecological record of key species and information on past use of the wider forest area revealed in this study, indicates how future management will require flexibility to maintain conservation ‘hotspots’.     

Fungal autolysate as a nutrient supplement for ethanol and chitosan production by <Emphasis Type="Italic">Mucor indicus</Emphasis>

Mucor indicus can be used to produce ethanol from a variety of sugars, including pentose’s. An extract of it, produced by autolysis, could replace yeast extract in culture medium with improved production of ethanol. At 10 g l⁻¹, the extract gave a higher ethanol yield (0.47 g g⁻¹) and productivity (0.71 g l⁻¹ h⁻¹) compared to medium containing yeast extract (yield 0.45 g g⁻¹; productivity 0.67 g l⁻¹ h⁻¹).     

On the extinction of the Dune Shearwater (Puffinus holeae) from the Canary Islands

Insular ecosystems have been subjected to severe hardship during the last millennia. Large numbers of insular bird species have undergone local disappearances and full extinctions, and a high number of insular birds are currently categorised as endangered species. In most of these cases, extinction—or endangerment—is in direct relation to the arrival of ‘aboriginal’ and/or imperialist waves of human settlement. Insular bird extinction events have been documented to have occurred at times corresponding to aboriginal settlement at many archipelagos and isolated islands, such as the Hawaiian Islands, New Zealand, the West Indies or the tropical Pacific Islands. However, no bird extinctions could be attributed to the first settlers of the Canary Islands—until now. The first accelerator mass spectrometer radiocarbon (¹⁴C) dating of collagen from a bone of the Dune Shearwater Puffinus holeae (3395 ± 30 year BP), an extinct bird from the Canary Islands, indicates a late Holocene extinction event. This relatively recent date, together with some features of this bird (large body size, breeding areas situated at very accessible places) and the absence of its bones from the entire archaeological record suggests that the extinction occurred close to the time that the first human settlement occurred on the islands.     

Biofuels: opportunities and challenges in India

Energy plays a vital role in the economic growth of any country. Current energy supplies in the world are unsustainable from environmental, economic, and societal standpoints. All over the world, governments have initiated the use of alternative sources of energy for ensuring energy security, generating employment, and mitigating CO₂ emissions. Biofuels have emerged as an ideal choice to meet these requirements. Huge investments in research and subsidies for production are the rule in most of the developed countries. India started its biofuel initiative in 2003. This initiative differs from other nations’ in its choice of raw material for biofuel production—molasses for bioethanol and nonedible oil for biodiesel. Cyclicality of sugar, molasses, and ethanol production resulted in a fuel ethanol program which suffered from inconsistent production and supply. The restrictive policies, availability of molasses, and cost hampered the fuel ethanol program. Inconsistent policies, availability of land, choice of nonnative crops, yield, and market price have been major impediments for biodiesel implementation. However, a coherent, consistent, and committed policy with long-term vision can sustain India’s biofuel effort. This will provide energy security, economic growth, and prosperity and ensure a higher quality of life for India.     

Rapid shoot regeneration in industrial ‘high starch’ sweetpotato (<Emphasis Type="Italic">Ipomoea batatas</Emphasis> L.) genotypes

Sweetpotato (Ipomoea batatas L.) is an important crop in North Carolina with annual production of 0.33 million tons, accounting for 37% of total US supply (USDA, Louisiana Farm Reporter 8(12), August 2008). To target industrial use, novel high-starch industrial-type varieties that contain more than 30% dry matter were developed by conventional breeding methods. In vitro cultures from selected genotypes were established using meristem culture. To establish regeneration procedures that could be coupled with transformation experiments, conditions for the induction of rapid shoot-organogenesis in leaf explants were compared using varying concentrations of the auxins ‘NAA’, ‘IAA’, ‘2,4-D’, and ‘4-FA’ either alone or in combination with zeatin riboside. Regeneration efficiencies, defined as the number of explants developing shoots out of the total number tested, were as high as 57% for the best genotypes, with a significant genotype-dependent response observed in all the hormone regimes evaluated. In all treatments, shoot regeneration was observed within 2 months. Our results led to the establishment of optimized in vitro regeneration procedures for the novel high-starch sweetpotato (SP) genotypes ‘DM01-158’, ‘FTA94’, ‘FT489’, and ‘PDM P4’ that are rapid and reliable.     

Target enrichment sequencing of 307 germplasm accessions identified ancestry of ancient and modern hybrids and signatures of adaptation and selection in sugarcane (Saccharum spp.), a ‘sweet’ crop with ‘bitter’ genomes

Sugarcane (Saccharum spp.) is a highly energy‐efficient crop primarily for sugar and bio‐ethanol production. Sugarcane genetics and cultivar improvement have been extremely challenging largely due to its complex genomes with high polyploidy levels. In this study, we deeply sequenced the coding regions of 307 sugarcane germplasm accessions. Nearly five million sequence variations were catalogued. The average of 98× sequence depth enabled different allele dosages of sequence variation to be differentiated in this polyploid collection. With selected high‐quality genome‐wide SNPs, we performed population genomic studies and environmental association analysis. Results illustrated that the ancient sugarcane hybrids, S. barberi and S. sinense, and modern sugarcane hybrids are significantly different in terms of genomic compositions, hybridization processes and their potential ancestry contributors. Linkage disequilibrium (LD) analysis showed a large extent of LD in sugarcane, with 962.4 Kbp, 2739.2 Kbp and 3573.6 Kbp for S. spontaneum, S. officinarum and modern S. hybrids respectively. Candidate selective sweep regions and genes were identified during domestication and historical selection processes of sugarcane in addition to genes associated with environmental variables at the original locations of the collection. This research provided an extensive amount of genomic resources for sugarcane community and the in‐depth population genomic analyses shed light on the breeding and evolution history of sugarcane, a highly polyploid species.     

Effects of a rapidly increasing population of common carp on vegetative cover and waterfowl in a recently restored Midwestern shallow lake

Although the common carp (Cyprinus carpio), an invasive benthic fish from Eurasia, has long been strongly implicated in the disappearance of vegetative cover and reduced waterfowl abundance in North American shallow lakes, the details of this relationship are obscure. This study documented ecological changes in a recently restored shallow lake (Hennepin and Hopper Lakes, IL, USA) at a time that it was experiencing a large increase in its carp population. We estimated the abundance and biomass of carp 7 years after this lake had been restored and then back-calculated carp population size across time while examining changes in the lake’s plant and waterfowl communities. We found that the biomass of carp remained below ~30 kg/ha for 5 years following restoration, but then increased to ~100 kg/ha in the sixth year following a strong recruitment event. Although a carp biomass of <30 kg/ha had no discernible effects on vegetative cover (which exceeded 90%) or waterfowl (which exceeded 150,000 individuals during fall censuses), the increase to 100 kg/ha was associated with a ~50% decrease in both vegetative cover and waterfowl. A further increase in carp biomass to over 250 kg/ha during the seventh year coincided with a decrease in the vegetative cover to 17% of the lake’s surface and a decline in waterfowl use to ~10% of its original value. These data suggest that the common carp is extremely damaging to the ecological integrity of shallow lakes when its density exceeds ~100 kg/ha. Since the biomass of carp in Midwestern shallow lakes commonly exceeds this value by 3–4 times, it seems likely that carp are responsible for the large-scale habitat deterioration described in many of these ecosystems. Handling editor: J. Cambray     

Effects of population density on forest structure and species richness and diversity of trees in Kampong Thom Province, Cambodia

This study examined differences in stand structure, tree species richness, and tree species diversity in relation to population density in Kampong Thom Province, Cambodia. Tree data were obtained from a 1997 forest inventory involving 60 clusters (540 plots) systematically distributed over 30% of the provincial forest area. Spatially referenced population data were obtained from the 1998 national population census. The average number of trees per cluster was 356/ha, the average basal area, 23 m²/ha, the average stand volume, 217 m³/ha, and the average aboveground biomass, 273 Mg/ha for all trees of DBH 10 cm and larger. The average species richness per cluster was 37 species, while average species diversity was measured as 0.916 using Simpson’s index and 2.98 by Shannon’s index. Significant negative correlations were generally found between population density surrounding clusters and tree density, basal area, stand volume, aboveground biomass, and species richness and diversity for three examined diameter classes (DBH of 10–30, ≥30, and ≥10 cm). As the distance from clusters for calculating population density increased, the correlation levels increased up to 5 or 7 km, depending on the variables and diameter class, and then stayed relatively constant for stand structure variables and decreased for species richness and diversity. The results indicate that evidence of disturbance was more pronounced at higher population density up to around 5 to 7 km. We suggest that introduction of greater controls on human disturbance should be a high priority for resource management and conservation in Kampong Thom Province and, presumably, Cambodia as a whole.     

Sugarcane improvement: how far can we go?

In recent years, efforts to improve sugarcane have focused on the development of biotechnology for this crop. It has become clear that sugarcane lacks tools for the biotechnological route of improvement and that the initial efforts in sequencing ESTs had limited impact for breeding. Until recently, the models used by breeders in statistical genetics approaches have been developed for diploid organisms, which are not ideal for a polyploid genome such as that of sugarcane. Breeding programs are dealing with decreasing yield gains. The contribution of multiple alleles to complex traits such as yield is a basic question underlining the breeding efforts that could only be addressed by the development of specific tools for this grass. However, functional genomics has progressed and gene expression profiling is leading to the definition of gene networks. The sequencing of the sugarcane genome, which is underway, will greatly contribute to numerous aspects of research on grasses. We expect that both the transgenic and the marker-assisted route for sugarcane improvement will contribute to increased sugar, stress tolerance, and higher yield and that the industry for years to come will be able to rely on sugarcane as the most productive energy crop.     

Fine-mapping of qRL6.1, a major QTL for root length of rice seedlings grown under a wide range of NH4 + concentrations in hydroponic conditions

Root system development is an important target for improving yield in cereal crops. Active root systems that can take up nutrients more efficiently are essential for enhancing grain yield. In this study, we attempted to identify quantitative trait loci (QTL) involved in root system development by measuring root length of rice seedlings grown in hydroponic culture. Reliable growth conditions for estimating the root length were first established to renew nutrient solutions daily and supply NH₄ ⁺ as a single nitrogen source. Thirty-eight chromosome segment substitution lines derived from a cross between ‘Koshihikari’, a japonica variety, and ‘Kasalath’, an indica variety, were used to detect QTL for seminal root length of seedlings grown in 5 or 500 μM NH₄ ⁺. Eight chromosomal regions were found to be involved in root elongation. Among them, the most effective QTL was detected on a ‘Kasalath’ segment of SL-218, which was localized to the long-arm of chromosome 6. The ‘Kasalath’ allele at this QTL, qRL6.1, greatly promoted root elongation under all NH₄ ⁺ concentrations tested. The genetic effect of this QTL was confirmed by analysis of the near-isogenic line (NIL) qRL6.1. The seminal root length of the NIL was 13.5–21.1% longer than that of ‘Koshihikari’ under different NH₄ ⁺ concentrations. Toward our goal of applying qRL6.1 in a molecular breeding program to enhance rice yield, a candidate genomic region of qRL6.1 was delimited within a 337 kb region in the ‘Nipponbare’ genome by means of progeny testing of F₂ plants/F₃ lines derived from a cross between SL-218 and ‘Koshihikari’.