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.     

甘蔗杂交品种核心种质重要农艺性状评价及亲缘关系分析

以107份甘蔗杂交品种核心种质为研究材料,ROC22为对照品种,通过新植和宿根2年的田间试验,对蔗茎公顷产量、甘蔗蔗糖分、公顷含糖量等性状进行最小显著差数法(LSD)两两比较和Duncan多重比较评价,筛选优异材料,利用SSR分子标记建立优异材料与我国骨干亲本的亲缘关系。结果表明36份材料在蔗茎公顷产量、甘蔗蔗糖分、公顷含糖量等指标上优于对照品种,可作为亲本进行杂交,且蔗茎公顷产量对公顷含糖量的影响远大于甘蔗蔗糖分对公顷含糖量的影响;20对SSR引物扩增得到292个标记,其中283个为多态性标记;优异材料与我国骨干亲本相似性系数范围在0.384~0.590之间,平均为0.437,存在较大的遗传差异;UPGMA聚类可将所有材料划分为5个类群,蔗茎公顷产量和公顷含糖量表现优异的材料在各类群都有分布,甘蔗蔗糖分表现优异的材料主要集中在Ⅱ、Ⅲ、Ⅳ类群,Ⅲ类群更集中;本研究为甘蔗种质创新、遗传育种提供了优异亲本材料,为杂交组合的配制提供重要指导。     

Calcium and Magnesium Levels in Agricultural Soil and Sewage Sludge in an Industrial Area from Southeastern Spain: Relationship with Plant (Saccharum officinarum) Disposition

This investigation was initiated to assess the following objectives: (1) to measure the total calcium and magnesium content in agricultural soil and sewage sludge from the Mediterranean coastal area of Motril (southeastern Spain); (2) to determine the pH values of indicated samples in order to evaluate first their influence on calcium and magnesium content, and second on levels of these minerals in the main crop (sugar cane: Saccharum officinarum) grown in the area; (3) to study the influence of industrial activities, first on calcium and magnesium cantents and pH values in agricultural soil, and second on calcium and magnesium concentrations present in sugar cane samples; (4) to check if the calcium and magnesium levels existing in agricultural soil exert any influence on corresponding element uptake by sugar cane plants. Calcium levels found in agricultural soil were significantly higher (p < 0.05) than those found in sewage sludge. Significant linear relationships between calcium and magnesium concentrations in agricultural soil and sewage sludge (p < 0.005) were found. Calcium concentrations found in soil from the industrialized area (39.2 ± 7.2 mg g^?1) were significantly higher than those corresponding to the non-industrialized area (31.0 ± 6.6 mg g^?1). The pH values determined in agricultural soil were significantly influenced by industrial activity (p < 0.05). The industrial activity and pH values measured in agricultural soil did not statistically influence either calcium or magnesium levels in the sugar cane plants. Calcium and magnesium concentrations existing in agricultural soil did not significantly influence the element uptake by sugar cane plants.     

Ethanol from Sugar Cane: Flask Experiments Using the EX-FERM Technique

Alcohol production at the laboratory scale from sugar cane pieces by the EX-FERM technique was studied with 37 strains of Saccharomyces spp. The EX-FERM process is novel in that it employs the simultaneous extraction and fermentation of the sucrose in a cane-water suspension. Two types of cane treatments were used: chips and shredded pith, either fresh or dried. A mother culture of the yeast was prepared in enriched cane juice and then added to the cane-water mixture. After static fermentation for 40 h at 30°C, the cane was removed, and fresh cane was added to the yeast-alcohol broth. After an additional 24 h, the cane was again removed and the liquor was analyzed. After the first 40-h cycle, sugar consumption was above 99% with 10 of the 37 yeast strains tested, and ethanol reached levels of 1.29 to 4.00 g per 100 ml, depending on the yeast strain. The final ethanol concentration reached 4.27 to 5.37 g per 100 ml, and sugar consumption was above 98% in three cases during a second EX-FERM cycle employing previously air-dried chips and pith. Product yields were within accepted values. Cane treatment did not appear to affect the results at this level.     

Biological nitrogen fixation associated with sugar cane

A recent¹⁵N dilution/N balance study confirmed that certain sugar cane varieties are capable of obtaining large contributions of nitrogen from plant-associated N₂ fixation. It was estimated that up to 60 to 80% of plant N could be derived from this source, and under good conditions of water and mineral nutrient supply, it may be possible to dispense with N fertilization of these varieties altogether. The recently discovered bacterium,Acetobacter diazotrophicus, apparently responsible for this N₂ fixation associated with the plants, has unique physiological properties for a diazotroph, such as tolerance to low pH, and high sugar and salt concentrations, lack of nitrate reductase, and nitrogenase activity which tolerates short-term exposure to ammonium. Furthermore, it also behaves as an endophyte, in that it is unable to infect sugar cane plants unless through damaged tissue or by means of VA mycorrhizae and is propagated via the planting material (stem pieces).     

Accumulation of recombinant cellobiohydrolase and endoglucanase in the leaves of mature transgenic sugar cane

A major strategic goal in making ethanol from lignocellulosic biomass a cost-competitive liquid transport fuel is to reduce the cost of production of cellulolytic enzymes that hydrolyse lignocellulosic substrates to fermentable sugars. Current production systems for these enzymes, namely microbes, are not economic. One way to substantially reduce production costs is to express cellulolytic enzymes in plants at levels that are high enough to hydrolyse lignocellulosic biomass. Sugar cane fibre (bagasse) is the most promising lignocellulosic feedstock for conversion to ethanol in the tropics and subtropics. Cellulolytic enzyme production in sugar cane will have a substantial impact on the economics of lignocellulosic ethanol production from bagasse. We therefore generated transgenic sugar cane accumulating three cellulolytic enzymes, fungal cellobiohydrolase I (CBH I), CBH II and bacterial endoglucanase (EG), in leaves using the maize PepC promoter as an alternative to maize Ubi1 for controlling transgene expression. Different subcellular targeting signals were shown to have a substantial impact on the accumulation of these enzymes; the CBHs and EG accumulated to higher levels when fused to a vacuolar-sorting determinant than to an endoplasmic reticulum-retention signal, while EG was produced in the largest amounts when fused to a chloroplast-targeting signal. These results are the first demonstration of the expression and accumulation of recombinant CBH I, CBH II and EG in sugar cane and represent a significant first step towards the optimization of cellulolytic enzyme expression in sugar cane for the economic production of lignocellulosic ethanol.     

Localization of Silicon in Specific Cell Wall Layers of the Stomatal Apparatus of Sugar Cane by Use of Energy Dispersive X-ray Analysis

By use of combined light, transmission electron, and scanningelectron microscopy with energy dispersive analysis of X-rays,silicon was localized in specific cell wall layers of the stomatalapparatus of sugar cane. The silicon occurs beneath the cuticlein layers of wall which correspond to the middle lamella andfirst-formed primary wall region of interior plant cells. Inother plants these regions are also reported high in pectinsand hemicellulose. It is suggested that the silicon may interactwith the pectins and hemicellulose and also with the ligninsand phenolics which may also be deposited in these wall layers. Saccharum officinarum L, sugar cane, stomata, silicon, X-ray analysis     

Life cycle assessment of cane-sugar on the island of mauritius

Goal, Scope and Background  Agricultural production includes not only crop production, but also food processing, transport, distribution, preparation, and disposal. The effects of all these must be considered and controlled if the food chain is to be made sustainable. The goal of this case study was to identify and review the significant areas of potential environmental impacts across the whole life cycle of cane sugar on the island of Mauritius. Methods  The functional unit was one tonne of exported raw sugar from the island. The life cycle investigated includes the stage of cane cultivation and harvest, cane burning, transport, fertilizer and herbicide manufacture, cane sugar manufacture and electricity generation from bagasse. Data was gathered from companies, factories, sugar statistics, databases and literature. Energy depletion, climate change, acidification, oxidant formation, nutrification, aquatic ecotoxicity and human toxicity were assessed. Results and Discussion  The inventory of the current sugar production system revealed that the production of one tonne of sugar requires, on average, a land area of 0.12 ha, the application of 0.84 kg of herbicides and 16.5 kg of N-fertilizer, use of 553 tons of water and 170 tonne-km of transport services. The total energy consumption is about 14235 MJ per tonne of sugar, of which fossil fuel consumption accounts for 1995 MJ and the rest is from renewable bagasse. 160 kg of CO₂ per tonne of sugar is released from fossil fuel energy use and the net avoided emissions of CO₂ on the island due to the use of bagasse as an energy source is 932,000 tonnes. 1.7 kg TSP, 1.21 kg SO₂,1.26 kgNO_xand 1.26 kg CO are emitted to the air per tonne of sugar produced. 1.7 kg N, 0.002 kg herbicide, 19.1 kg COD, 13.1 kgTSS and 0.37 kg PO₄ ^3- are emitted to water per tonne of sugar produced. Cane cultivation and harvest accounts for the largest environmental impact (44%) followed by fertilizer and herbicide manufacture (22%), sugar processing and electricity generation (20%), transportation (13%) and cane burning (1%). Nutrification is the main impact followed by acidification and energy depletion. Conclusions  There are a number of options for improvement of the environmental performance of the cane-sugar production chain. Cane cultivation, and fertilizer and herbicide manufacture, were hotspots for most of the impact categories investigated. Better irrigation systems, precision farming, optimal use of herbicides, centralisation of sugar factories, implementation of co-generation projects and pollution control during manufacturing and bagasse burning are measures that would considerably decrease resource use and environmental impacts. Recommendation and Outlook  LCA was shown to be a valuable tool to assess the environmental impacts throughout the food production chain and to evaluate government policies on agricultural production systems.     

Biological Nitrogen Fixation in Sugar Cane: A Key to Energetically Viable Biofuel Production

The advantages of producing biofuels to replace fossil energy sources are derived from the fact that the energy accumulated in the biomass is captured directly from photosynthesis and is thus renewable, and that the cycle of carbon dioxide fixation by the crop, followed by burning of the fuel makes no overall contribution to atmospheric CO₂ or, consequently, to global warming. However, these advantages are negated if large quantities of fossil fuels need to be used to grow or process the biofuel crop. In this regard, the Brazilian bioethanol program, based on the fermentation/distillation of sugar cane juice, is particularly favorable, not only because the crop is principally hand harvested, but also because of the low nitrogen fertilizer use on sugar cane in Brazil. Recent ¹⁵N and N balance studies have shown that in some Brazilian cane varieties, high yields are possible without N fertilization because the plants are able to obtain large contributions of nitrogen from plant-associated biological N₂ fixation (BNF). The N₂-fixing acid-tolerant bacterium Acetobacter diazotrophicus was first found to occur within roots, stems, and leaves of sugar cane. Subsequently, two species of Herbaspirillum also have been found to occur within the interior of all sugar cane tissues. The discovery of these, and other N₂-fixing bacteria that survive poorly in soil but thrive within plant tissue (endophytic bacteria), may account for the high BNF contributions observed in sugar cane. Further study of this system should allow the gradual elimination of N fertilizer use on sugar cane, at least in Brazil, and opens up the possibility of the extension of this efficient N₂-fixing system to cereal and other crops with consequent immense potential benefits to tropical agriculture.     

Infection and Colonization of Sugar Cane and Other Graminaceous Plants by Endophytic Diazotrophs

Agriculturally important grasses such as sugar cane (Saccharum sp.), rice (Oryza sativa), wheat (Triticum aestivum) sorghum (Sorghum bicolor), maize (Zea mays), Panicum maximum, Brachiaria spp., and Pennisetum purpureum contain numerous diazotrophic bacteria, such as, Acetobacter diazotrophicus, Herbaspirillum spp., Azospirillum spp. These bacteria do not usually cause disease symptoms in the plants with which they are associated and the more numerous of them, for example, Herbaspirillum spp. and A. diazotrophicus, are obligate or facultative endo-phytes that do not survive well (or at all) in native soil; these are thought to be spread from plant generation to plant generation via seeds, vegetative propagation, dead plant material, and possibly by insect sap feeders. By contrast, Azospirillum spp. are not wholly endophytic but are root-associated, soil-dwelling bacteria that are also often found within plants, probably entering host plants via seeds or via wounds/cracks at lateral root junctions. Endophytic diazotrophs have been isolated from a number of grasses in which significant biological N₂ fixation (BNF) has been demonstrated, particularly Brazilian sugar cane varieties, but also in rice, maize, and sorghum. However, although the endophytic diazotrophs are held to be the causative agents of the observed BNF, direct evidence for this is lacking. Therefore, in this review we examine probable sites of bacterial multiplication and/or BNF within endophyte-containing grasses and discuss these in terms of potential benefits (or not) to both host plants and bacteria. In particular, we examine how potentially large numbers of bacteria, especially Herbaspirillum spp., A. diazotrophicus, and Azospirillum spp., can exist extracellularly within non-specialized (for symbiotic purposes) regions such as xylem vessels and intercellular spaces. The processes of infection and colonization of various grasses (particularly sugar cane) by diazotrophic endophytes are also described, and these are compared with those of important (nondiazotrophic) endophytic sugar cane pathogens such as Clavibacter xyli subsp. xyli and Xanthomonas albilineans.     

Evaluation of sugar feedstocks for bio-based chemicals: A consequential,regionalized life cycle assessment

Fermentable sugars are an attractive feedstock for the production of bio-based chemicals. However, little is known about the environmental performance of sugar feedstocks when demand for sugars increases, and when local conditions and sensitivities of receiving ecosystems are taken into account. Production of monosaccharides from various first- and second-generation feedstocks (sugar beet, sugar cane, wheat, maize, wood, residual woodchips, and sawdust) in different geographic locations was assessed and compared as feedstock for monoethylene glycol (MEG) using consequential, regionalized life cycle assessment. Sugar cane grown in Thailand performed best in all three areas of protection, that is, for life cycle impacts on human health, ecosystem quality, and resources (respectively, equal to −7.6 × 10⁻⁵ disability-adjusted life years, −1.2 × 10⁻⁸ species-years and −0.046 US dollars per amount of feedstock needed to produce 1 kg of MEG). This was mainly due to benefits from by-products—incineration of sugar cane bagasse generating electricity and use of sugar cane molasses for the production of bioethanol. The wood-based feedstocks and maize performed worse than sugar cane and sugar beet, but their evaluation did not consider that sugar extraction technology from lignocellulose is immature, while identification of marginal suppliers of the marginal crop is particularly uncertain for maize. Wheat grown in Russia performed the worst mainly due to low agricultural yields (with impacts equal to 8.9 × 10⁻⁵ disability-adjusted life years, 6.9 × 10⁻⁷ species-years, and 1.8 US dollars per amount of feedstock required to produce 1 kg of bio-based MEG). Our results suggest that selection of sugar feedstocks for bio-based chemicals should focus on (i) the intended use of by-products and functions they replace and (ii) consideration of geographic differences in parameters that influence life cycle inventories, while spatial differentiation in the life cycle impact assessment was less influential.     

Ethanol from sugar crops

Soluble sugars, like sucrose, glucose and fructose, are transformed by yeast into ethanol and carbon dioxide. These sugars are stored in photosynthetically efficient plants like sugar cane. Recent developments in the transformation of sucrose present in sugar cane or sweet sorghum into ethanol, include the use of the Tilby machine, a high-temperature extraction process and the Ex-Ferm process. This review covers kinetic aspects of ethanol production, yeast immobilization techniques, yeast properties and fermentation byproducts.     

Influence of nitrogen fertilisation on the population of diazotrophic bacteria Herbaspirillum spp. and Acetobacter diazotrophicus in sugar cane (Saccharum spp.)

We report studies on the possible effects of fertilisation with high level of N (300 kg of N ha^-1) on the occurrence and numbers of the diazotrophic bacteria Herbaspirillum spp. and Acetobacter diazotrophicusin sugar cane plants. In the sugar cane genotype SP79-2312, the N fertilised plants generally showed higher concentrations of this element. These same plants also had lower numbers of A. diazotrophicus, while the population of Herbaspirillum spp. was not affected by N application. These differences in the concentration of N and the numbers of A. diazotrophicus due to N application were not shown in the variety SP70-1143. The numbers of A. diazotrophicus were also shown to be influenced by the harvest time, becoming reduced in the harvests that coincided with dry periods of the year.     

Response of eight sugarcane cultivars to glyphosine and glyphosate ripeners

Studies were conducted on eight sugarcane (Saccharum spp. hydrid) cultivars during the 1982–83 (plant crop) and 1983–84 (ratoon crop) growing seasons to determine the effects of glyphosine (Polaris) (N,N-bis (phosphonomethyl) glycine) and glyphosate (Polado) (sodium-N-(phosphonomethyl) glycine) on stalk sucrose content and yield. Difference due to crops (plant vs. ratoon) for sugarcane quality, kilograms of sugar per ton of cane (S/T), sugarcane yield, tons of cane per hectare (TCH), and sugar yield, tons of sugar per hectare (TSH) were significant. Significant differences were found in quality for the ratoon crop and cane and sugar yield in both crops due to ripener treatment. Cultivars in both crops differed significantly in quality and yield. Harvest dates were significantly different for all plant characteristics. Interactions of cultivar by treatment for the plant crop, harvest date by treatment for the ratoon crop, and cultivar by harvest date for both crops for cane quality also were significant. Time from ripener application to achievement of maximum sugar concentration also depended on cultivar. This is important in determining the economic benefits of a ripener treatment. Climatic conditions may also affect the benefits of such applications.Mention of trade name or proprietary product does not imply or constitute an endorsement or recommendation by the USDA or the University of Florida.     

Response of eight sugarcane cultivars to glyphosine and glyphosate ripeners

Studies were conducted on eight sugarcane (Saccharum spp. hydrid) cultivars during the 1982–83 (plant crop) and 1983–84 (ratoon crop) growing seasons to determine the effects of glyphosine (Polaris) (N,N-bis (phosphonomethyl) glycine) and glyphosate (Polado) (sodium-N-(phosphonomethyl) glycine) on stalk sucrose content and yield. Difference due to crops (plant vs. ratoon) for sugarcane quality, kilograms of sugar per ton of cane (S/T), sugarcane yield, tons of cane per hectare (TCH), and sugar yield, tons of sugar per hectare (TSH) were significant. Significant differences were found in quality for the ratoon crop and cane and sugar yield in both crops due to ripener treatment. Cultivars in both crops differed significantly in quality and yield. Harvest dates were significantly different for all plant characteristics. Interactions of cultivar by treatment for the plant crop, harvest date by treatment for the ratoon crop, and cultivar by harvest date for both crops for cane quality also were significant. Time from ripener application to achievement of maximum sugar concentration also depended on cultivar. This is important in determining the economic benefits of a ripener treatment. Climatic conditions may also affect the benefits of such applications.     

Determination of endotoxins in sugar with the Limulus test.

The Limulus amebocyte lysate test has been used for determination of pyrogens in sugar of different qualities. All the samples of domestic white sugar and beet raw sugar produced in Sweden during 1976 had a very low content of endotoxins, less than 10 ng/g of sugar. Imported cane raw sugar was, however, highly contaminated. The highest value obtained corresponds to about 100 mg of Escherichia coli endotoxin per g of raw sugar. Such crude sugar cannot, even after refining, be used for medical purposes. Instead, Swedish beet sugar is used as the raw material for production of invert sugar solutions for parenteral administration. The amount of endotoxin in this sugar is less than 1 ng/g.     

RNAi-directed downregulation of betaine aldehyde dehydrogenase 1 (OsBADH1) results in decreased stress tolerance and increased oxidative markers without affecting glycine betaine biosynthesis in rice (Oryza sativa)

Chemically inducible gene switches can provide precise control over gene expression, enabling more specific analyses of gene function and expanding the plant biotechnology toolkit beyond traditional constitutive expression systems. The alc gene expression system is one of the most promising chemically inducible gene switches in plants because of its potential in both fundamental research and commercial biotechnology applications. However, there are no published reports demonstrating that this versatile gene switch is functional in transgenic monocotyledonous plants, which include some of the most important agricultural crops. We found that the original alc gene switch was ineffective in the monocotyledonous plant sugar cane, and describe a modified alc system that is functional in this globally significant crop. A promoter consisting of tandem copies of the ethanol receptor inverted repeat binding site, in combination with a minimal promoter sequence, was sufficient to give enhanced sensitivity and significantly higher levels of ethanol inducible gene expression. A longer CaMV 35S minimal promoter than was used in the original alc gene switch also substantially improved ethanol inducibility. Treating the roots with ethanol effectively induced the modified alc system in sugar cane leaves and stem, while an aerial spray was relatively ineffective. The extension of this chemically inducible gene expression system to sugar cane opens the door to new opportunities for basic research and crop biotechnology.     

Determination of endotoxins in sugar with the Limulus test

The Limulus amebocyte lysate test has been used for determination of pyrogens in sugar of different qualities. All the samples of domestic white sugar and beet raw sugar produced in Sweden during 1976 had a very low content of endotoxins, less than 10 ng/g of sugar. Imported cane raw sugar was, however, highly contaminated. The highest value obtained corresponds to about 100 mg of Escherichia coli endotoxin per g of raw sugar. Such crude sugar cannot, even after refining, be used for medical purposes. Instead, Swedish beet sugar is used as the raw material for production of invert sugar solutions for parenteral administration. The amount of endotoxin in this sugar is less than 1 ng/g.     

Infection of sugar cane by the nitrogen-fixing bacterium Acetobacter diazotrophicus

Significant nitrogen fixation has recently been demonstratedin Brazilian sugar cane (Saccharum officinarum) cultivars knownto form associations with a number of diazotrophs, includingAcetobacter diazotrophicus, an acid-tolerant endophytic bacteriumwhich grows best on a sucrose-rich medium. In a series of experiments,aseptically-grown sugar cane plantlets were rooted in a liquidmedium and inoculated with A. diazotrophicus originally isolatedfrom field-grown sugar cane. After 4, 7, 9, and 15 d, plantswere examined under light, scanning and transmission electronmicroscopes and the presence of A. diazotrophicus on and withinplant tissues was confirmed by immunogold labelling. By 15 d,external bacterial colonization was seen on roots and lowerstems, particularly at cavities in lateral root junctions. Theloose cells of the root cap at root tips were a site of entryof the bacteria into root tissues. Both at lateral root junctionsand root tips, bacteria were also seen in enlarged, apparentlyintact, epidermal cells. After 15 d, bacteria were present inxylem vessels at the base of the stem, many connected via mucusto spiral secondary thickening. There was no obvious pathogenicreaction to the bacteria within the xylem. From these observations,it is proposed that, under experimental conditions, A. diazotrophicusfirstly colonized the root and lower stem epidermal surfacesand then used root tips and lateral root junctions to enterthe sugar cane plant where it was distributed around the plantin the transpiration stream. It is further suggested that thexylem vessels in the dense shoots of mature plants are alsoa possible site of N₂-fixation by diazotrophs as they providethe low pO₂ and energy as sucrose necessary for nitrogenaseactivity. Key words: Acetobacter diazotrophicus, endophyte, infection, nitrogen fixation, sugar cane.