Extraction of polymeric galactoglucomannans from spruce wood by pressurized hot water

The major type of non-cellulosic polysaccharides (hemicelluloses) in softwoods, the partly acetylated galactoglucomannans (GGMs), which comprise about 15% of spruce wood, have attracted growing interest because of their potential to become high-value products with applications in many areas. The main objective of this work was to explore the possibilities to extract galactoglucomannans in native, polymeric form in high yield from spruce wood with pressurised hot-water, and to obtain a deeper understanding of the process chemistry involved. Spruce (Picea abies) chips and ground wood particles were extracted using an accelerated solvent extractor (ASE) in the temperature range 160 – 180°C. Detailed chemical analyses were done on both the water extracts and the wood residues. As much as 80 – 90% of the GGMs in spruce wood, i.e. about 13% based on the original wood, could be extracted from ground spruce wood with pure water at 170 – 180°C with an extraction time of 60 min. GGMs comprised about 75% of the extracted carbohydrates and about 60% of the total dissolved solids. Other substances in the water extracts were xylans, arabinogalactans, pectins, lignin and acetic acid. The yields from chips were only about 60% of that from ground wood. Both the GGMs and other non-cellulosic polysaccharides were extensively hydrolysed at severe extraction conditions when pH dropped to the level of 3.5. Addition of sodium bicarbonate increased the yields of polymeric GGMs at low additions, 2.5 – 5 mM, where the end pH remained around 3.9. However, at higher addition levels the yields decreased, mainly because the acetyl groups in GGMs were split off, leading to a low solubility of GGMs. Extraction with buffered water in the pH range 3.8 – 4.4 gave similar yields as with plain water, but gave a higher yield of polymeric GGMs. Moreover, at these pH levels the hydrolysis of acetyl groups in GGMs was significantly inhibited. It was concluded that hot-water extraction of polymeric GGMs in good yields (up to 8% of wood) demands appropriate control of pH, in a narrow range about 4. These results were supported by a study of hydrolysis of GGM at constant pH in the range of 3.8 – 4.2 where a kinetic model for degradation of GGM was developed. The influence of wood particle size on hot-water extraction was studied with particles in the range of 0.1 – 2 mm. The smallest particles (< 0.1 mm) gave 20 – 40% higher total yield than the coarsest particles (1.25 – 2 mm). The difference was greatest at short extraction times. The results indicated that extraction of GGMs and other polysaccharides is limited mainly by the mass transfer in the fibre wall, and for coarse wood particles also in the wood matrix. Spruce sapwood, heartwood and thermomechnical pulp were also compared, but only small differences in yields and composition of extracts were found. Two methods for isolation and purification of polymeric GGMs, i.e. membrane filtration and precipitation in ethanol-water, were compared. Filtration through a series of membranes with different pore sizes separated GGMs of different molar masses, from polymers to oligomers. Polysaccharides with molar mass higher than 4 kDa were precipitated in ethanol-water. GGMs comprised about 80% of the precipitated polysaccharides. Other polysaccharides were mainly arabinoglucuronoxylans and pectins. The ethanol-precipitated GGMs were by 13C NMR spectroscopy verified to be very similar to GGMs extracted from spruce wood in low yield at a much lower temperature, 90°C. The obtained large body of experimental data could be utilised for further kinetic and economic calculations to optimise technical hot-water extractionof softwoods.

Corn yield response to weed and fall armyworm controls

The interference imposed the by weeds on corn decreases practically all vegetative characteristics. As consequence, the green ear and grain yield are also reduced. Losses due to the fall armyworm (Spodoptera frugiperda) attack can reduce corn grain yield up to 34%. In general, weed and insect control issues are addressed separately in research papers. Nevertheless, interaction between weeds and insects may exist. This study aimed to evaluate green ear and corn grain yield response to weed and fall armyworm control. A completely randomized block design with split-plots and five replicates was adopted. Corn cultivar AG 1051 was grown under weedy conditions or with control by hand hoeings performed at 20 and 40 days after planting. Fall armyworm control (applied to subplots) was performed with sprays of water (control), deltamethrin (5g active ingredient ha-1); neem oil, at 0.5% (diluted in water), and neem leaf extract at 5%. Each product was sprayed three times, at seven-day intervals, starting at the 7th day after planting, using 150 L ha-1 of the tank solution. Dry mass of the above-ground part, internode diameter, leaf length, leaf width, leaf area, green ear yield and grain yield of corn were reduced due to the lack of weed control. Fall armyworm control in the weeded plots did not influence green ear yield and grain yield, except green mass of marketable, husked ears, which was reduced when the caterpillar was not controlled. Without weed control, neem extracts and deltamethrin sprays provided highest yields of number and total weight of green ears with husks, number and weight of marketable ears with husks and number of marketable ears without husks. The best results for husked ear mass and for grain yield were obtained with neem extract and deltamethrin, respectively.

Production of Chemicals from Woody Biomass by Wet Oxidation

The production of chemicals from sawdust by wet oxidation has been investigated. Two different concentrations of sawdust; 54054 mg/l and 32683 mg/l were used in the study. The wet oxidation operating conditions were; 175 deg.C – 225 deg.C, 1MPa Oxygen, and 40 minutes to 120 minutes reaction time. Carboxylic acids were among the chemicals produced in the process. The total yield of carboxylic acids was found to increase with temperature. Also, higher yields of carboxylic acids were observed at a lower sawdust concentration. This was probably due to the high oxygen-biomass ratio at lower sawdust concentration. Higher oxygen availability at low sawdust concentration resulted in increased conversion of the sawdust; hence the higher yields of carboxylic acids. At lower sawdust concentration, a total carboxylic acid yield of 25.59 wt% was attained at 200 deg.C and 40 minutes reaction time. At higher sawdust concentration, a total carboxylic acid yield of 15.57 wt% was attained at 200 deg.C and 40-minutes reaction time. The carboxylic acids identified include formic acid, acetic acid, succinic acid and oxalic acid. The optimum temperature for the production of formic acid was found to be 200 deg.C, while the optimum temperature for the production of acetic acid was found to be 225 deg.C. A temperature of 225 deg.C and relatively short reaction time of 10 minutes was found to be the optimal condition for the production of succinic acid. Formic acid was produced in the highest yield, with an optimal yield of 13.69wt %, when the reaction temperature and time are 200 deg.C and 40 minutes respectively. The yield of formic acid was found to decrease significantly when further increasing the temperature to 225 deg.C. This was presumably due to thermal decomposition of formic acid at relatively higher temperature. However, the yield of acetic acid was found to steadily increase with temperature. This is because acetic is more thermally stable than formic acid. The yield of acetic acid did not decrease after the temperature was increased to 225 deg.C. Optimal yield of acetic acid (7.98wt %) was achieved at; 225 deg.C, and 40 minutes reaction time. Succinic acid was produced only at temperatures of 200 deg.C and 225 deg.C. Optimal yield of succinic acid (5.66wt %) was attained under the following conditions; 32683 mg/l, 225 deg.C, 1MPa O2, and 10-minutes reaction time. Oxalic acid was produced in the lowest yield and, less frequently. The optimal yield of oxalic acid (4.02 wt%) was attained at 175 deg.C and 80-minutes of reaction time The Total Organic Carbon (TOC) is found to be higher when increasing the operating temperature, thus suggesting that more organic compounds are formed at higher temperatures. The identified carboxylic acids could only account for less than 30% of the measured COD content of the various wet oxidation samples. This implies that some other unidentified compounds (reaction products) must have been present. In general, wet oxidation seems to be an effective method for converting lignocellulosic biomass into useful chemicals. Relatively higher temperatures have been found to favor the production of carboxylic acids from sawdust.

Topochemistry of physical and chemical pretreatments of biomass as investigated by FE-SEM, XPS and ToF-SIMS

Surface chemistry is of great importance in plant biomass engineering and applications. The surface chemical composition of biomass which includes lignin, carbohydrates and extractives influences its interactions with chemical agents, such as pulp processing/papermaking chemicals, or enzymes for different purposes. In this thesis, the changes in the surface chemical composition of lignocellulosic biomass after physical modification for the improvement of resulting paper properties and chemical treatment for the enhancement of enzymatic hydrolysis were investigated. Low consistency (LC) refining was used as physical treatment of bleached softwood and hardwood pulp samples, and the surface chemistry of refined samples was investigated. The refined pulp was analysed as whole pulp while the fines-free fibre samples were characterized separately. The fines produced in LCrefining contributed to an enlarged surface specific area as well as the change of surface coverage by lignin and extractives, as investigated by X-ray photoelectron spectroscopy (XPS). The surface coverage by lignin of the whole pulp decreased after refining while the surface coverage by extractives increased both for pine and eucalyptus. In the case of pine, the removal of fines resulted in reduction of the surface coverage by extractives, while the surface coverage by lignin increased on fibre sample (without fines). In the case of eucalyptus, the surface coverage by lignin of fibre samples decreased after the removal of fines. In addition, the surface distribution of carbohydrates, lignin and extractives of pine and eucalyptus samples was determined by Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS). LC-refining increased the amounts of pentose, hexose and extractives on the surface of pine samples. ToF-SIMS also gave clear evidence about xylan deposition and reduction of surface lignin distribution on the fibre of eucalyptus. However, the changes in the surface chemical composition during the physical treatment has led to an increase in the adsorption of fluorescent whitening agents (FWAs) on fibres due to a combination of electro-static forces, specific surface area of fibres and hydrophobic interactions. Various physicochemical pretreatments were conducted on wood and non-wood biomass for enhancing enzymatic hydrolysis of polysaccharides, and the surface chemistry of the pretreated and enzymatically hydrolysed samples was investigated by field emission scanning electron microscopy (FE-SEM), XPS and ToF-SIMS. A hydrotrope was used as a relatively novel pretreatment technology both in the case of wood and non-wood biomass. For comparison, ionic liquid and hydrothermal pretreatments were applied on softwood and hardwood as well. Thus, XPS analysis showed that the surface lignin was more efficiently removed by hydrotropic pretreatment compared to ionic liquid or hydrothermal pretreatments. SEM analysis also found that already at room temperature the ionic liquid pretreatments were more effective in swelling the fibres compared with hydrotropic pretreatment at elevated temperatures. The enzymatic hydrolysis yield of hardwood was enhanced due to the decrease in surface coverage of lignin, which was induced by hydrotropic treatment. However, hydrotropic pretreatment was not appropriate for softwood because of the predominance of guaiacyl lignin structure in this material. In addition, the reduction of surface lignin and xylan during pretreatment and subsequent increase in cellulose hydrolysis by enzyme could be observed from ToF-SIMS results. The characterisation of the non-wood biomass (e.g. sugarcane bagasse and common reed) treated by hydrotropic method, alkaline and alkaline hydrogen peroxide pretreatments were carried out by XPS and ToF-SIMS. According to the results, the action for the removal of the surface lignin of non-wood biomass by hydrotropic pretreatment was more significant compared to alkaline and alkaline hydrogen peroxide pretreatments, although a higher total amount of lignin could be removed by alkaline and alkaline hydrogen peroxide pretreatment. Furthermore, xylan could be remarkably more efficiently removed by hydrotropic method. Therefore, the glucan yield achieved from hydrotropic treated sample was higher than that from samples treated with alkaline or alkaline hydrogen peroxide. Through the use of ToF-SIMS, the distribution and localization of lignin and carbohydrates on the surface of ignocelluloses during pretreatment and enzymatic hydrolysis could be detected, and xylan degradation during enzymatic hydrolysis could also be assessed. Thus, based on the results from XPS and ToF-SIMS, the mechanism of the hydrotropic pretreatment in improving the accessibility of enzymes to fibre and further ameliorating of the enzymatic saccharification could be better elucidated.

The effects of pigweed redroot (Amaranthus retoflexus) weed competition and its economic thresholds in corn (Zea mays)

The aim of this study was to determine the economic damage threshold of Pigweed redroot for corn regarding its density. An experiment was conducted at the Agriculture Research station of Islamic Azad University branch of Gonabad during 2006. The experiment was carried out as a factorial in a randomized complete block design with three replications. In the experiments, the factors included corn (var. 704) densities of 7.5, 8.5 and 9.5 plants m-2 and pigweed redroot densities of 0, 2, 4, 6 and 8 plants m-2. The increase in Pigweed redroot density, decrease in crop grain and biomass yield components such as ear length, ear diameter, number of grains per row, row number, grain number in ear, grain yield and biological yield of corn, decreased. Also, with an increase in corn density, the number of grain per rows, row number, grain yield and biological yield of corn increased. The economic thresholds density of Pigweed redroot was 0.09 to 0.13 plants m-2 in corn different densities, and increased with corn density increases.

Effect of weed interference on soybean cultivars at two sowing times

This study aimed to evaluate the effects of weed interference on soybean cultivars at two sowing times in Urutaí, GO. The treatments were arranged in split-plots, and the sowing times (November 16 and December 16, 2009) were allocated in the plots; the soybean cultivars [BRSGO Amaralina, P98C81 (semi-late cycle) BRSGO Raissa, BRSGO Indiara, P98Y11 (median cycle) and BRSGO 7560, BRSGO Caiapônia, Emgopa 302RR (early cycle)] were allocated in the split-plots; and the coexistence or non coexistence of soybean cultivars with weeds, throughout their cycle, was allocated in the split-plots. Non coexistence was established by manual weeding. The experiment was arranged in randomized blocks with four replications. It was verified that the optimal time for sowing soybeans was the month of November, and that under these conditions, the cultivars had higher competitive ability against weeds. Late sowing affected the cycle, development, and yield of the soybean cultivars; this effect was greater under the influence of the weed community.

Competitive ability of soybean cultivars with horseweed (Conyza bonariensis)

The existence of large areas infested with populations of Conyza spp. resistant to glyphosate in Brazil demands appropriate and integrated management strategies. This experiment aimed to identify soybean cultivars with greater competitive ability with horseweed plants and to determine plant characteristics associated with this ability. The experiment was arranged in a randomized complete block design with split plots. Seven soybean cultivars (CD 225 RR, BRS 232, CD 226 RR, NK 7054 RR, BMX Apollo RR, BRS 245 RR and BRS 255 RR) were allocated in the plots, and two interference situations (absence and 13.3 plants of Conyza m-2, transplanted seven days before soybean planting) in the subplots. The average yield loss due to competition with horseweed was 25%. Cultivar CD 226 RR showed no significant grain yield loss due to competition, compared to the control without infestation, but showed the lowest average grain yield. The BRS 232 genotype showed loss of grain yield of only 14%, and presented positive plant height and leaf mass at 20 DAE, as well as dry matter of stems+branches in all evaluations, features related to its higher performance and greater ability to withstand competition with horseweed plants.

Density and sowing season of two Brachiaria species on the soybean culture

The aim of this study was to evaluate the growth and yield of soybean cultivar M-8766 in consortium with Brachiaria brizantha. BRS Piata and Brachiaria ruziziensis at different densities and sowing dates. The experimental design was randomized blocks with treatments arranged in a factorial 2 x 2 x 3 with four replications. Used as factors grass species (Brachiaria brizantha Piata and Brachiaria ruziziensis BRS) intercropped with soybean M-8766, sowing dates (12 and 24 days after soybean emergence) and three seeding rates (0, 5, 10 kg ha-1 of seed). At 71 days after soybean emergence were evaluated plant height, stem diameter, dry mass of leaves, stems and shoots, and 4 months after sowing determined the weight of 100 grains and soybean yield. The results showed that when seeded at a density of 10 kg ha-1 at 12 and 24 DAE soy, Brachiaria brizantha. BRS Piata caused reduction in yield in the order of 6.71% and 3.03% respectively, while the Brachiaria ruziziensis was one that caused a greater reduction in productivity in the order of 13.42 and 16.23%, respectively, of these values expression when considering the price of soybean sack. B. ruziziensis expressed less competitive with soybean. However, the large biomass production of this grass provides deployment system till the next harvest.

Assessing the Competitive Ability of Rhynchosia capitata; an Emerging Summer Weed in Asia

Rhynchosia capitata is a newly emerging threatening weed of summer crops in many Asian countries. We conducted 2-yr experiments to evaluate R. capitata competition with mungbean under irrigated conditions. Rhynchosia capitata was allowed to compete with mungbean for 0, 3, 4, 5, 6, 7 weeks after planting and for full crop season. The competitive ability of R. capitata was assessed by measuring its dry weight, micro and macro nutrient contents and uptake; and its effects on mungbean growth and yield parameters. The results showed that full season weed competition produced highest dry weight of R. capitata and its macro and micronutrient contents and uptake. Yield and yield components of mungbean decreased in a linear fashion with increase in competition period of Rhynchosia capitata. Full season competition of R. capitata reduced the seed yield of mungbean by 49% and 46% during 2011 and 2012 respectively. In conclusion, damaging effect of R. capitata intrusion on mungbean yield is associated with duration of its presence in crop, accumulation of dry matter and the nutrient uptake by R. capitata, which otherwise should be available to crop.

Purification and characterization of a phytoalexin elicitor from spores of the saprobe Mucor ramosissimus

Plants accumulate antimicrobial compounds (phytoalexins) in response to a wide variety of microorganisms. Mucor ramosissimus Samutsevitsch is a saprobe capable of inducing phytoalexin production in soybean cotyledons and in the leaves of tropical Rubiaceae on whose surface it has been found. In the present study, the elicitor from M. ramosissimus was partially purified and the activity compared to that of a glucan elicitor isolated from Phytophthora sojae. Optimal isolation of the elicitor (based on fungal growth, yield of spores and elicitor activity) was achieved by autoclaving spores obtained from nine day-old cultures of the fungus. The elicitor was precipitated with ethanol and purified by chromatography on an anion exchange column, which retained the elicitor, and a Concanavalin A-affinity matrix, to which the elicitor did not bind. The purification resulted in a considerable increase (six-fold) in the specific activity of the elicitor. Neutral sugar composition, analyzed by HPLC, revealed the predominance of mannose, followed by glucose and galactose, whereas colorimetric quantification showed the presence of uronic acids. GC-MS analysis of the elicitor revealed the predominance of glucuronic acid and mannose. These results suggest that fragments of mucoran-type polysaccharides are the phytoalexin elicitors present in the spores of the saprobe M. ramosissimus. Our results also indicate for the first time that soybean cotyledon tissues can recognize fragments of glucuronic-acid heteropolymers as phytoalexin elicitors.

Synecological comparisons sustained by ecophysiological fingerprinting of intrinsic photosynthetic capacity of plants as assessed by measurements of light response curves

In some literature variations in photosynthetic rates are considered to be of little relevance for individual fitness. This depends among other things on how one defines fitness, i.e. if one takes strictly Darwinian fitness as seed production or if one needs to evaluate particular traits and consider plant establishment. It also matters if one takes the Darwinian "organism individual" as the central entity in evolution ("individual fitness") or the "species individual" in a modified "Structure of Evolutionary Theory" sensu Stephen Jay Gould. A phenotypically expressed trait like photosynthetic rate, even if intra- and interspecific differences may be small, can matter in habitat performance and niche acquisition. Light dependence curves (LCs) of photosynthetic rates are now readily measured under field conditions using miniaturized equipment of pulse amplitude modulated fluorometers. In contrast to actual momentary measurements of quantum yield of photosynthesis under actually prevailing ambient conditions, LC measurements reflect the expressed intrinsic capacity of photosynthesis. In this review we explore the power of LC measurements yielding cardinal points such as maximum apparent electron transport rate of photosystem II (ETRmax) and saturating photosynthetically active radiation (PARsat) in making intra- and interspecific comparisons of plant performance and synecological fingerprinting in ecophysiological studies across species, sites, habitats and ecosystems.

Photosynthetic properties of three Brazilian seaweeds

Photosynthetic performance of distinct marine macroalgae, Ulva fasciata Delile (green alga), Lobophora variegata (J. V. Lamouroux) Womersley ex E. C. Oliveira (brown alga), and Plocamium brasiliensis (Greville) M. A. Howe & W. R. Taylor (red alga), were compared using a pulse amplitude-modulated fluorometer. The maximum quantum yield (Fv/Fm) ranged from 0.80 to 0.51, and the lowest value was found in P. brasiliensis. Under 400 µmol photons m-2 s-1 irradiance, the highest value of photochemical quenching (qP = 0.92 ± 0.13) was observed for U. fasciata. The red alga P. brasiliensis dissipated high amounts of excitation energy (qN = 0.56 ± 0.09), resulting in relatively low values for the effective quantum yield of PS-II (0.23 ± 0.04), as well as for the relative electron transport rate (3.3 ± 0.7). The high photosynthetic potential found for U. fasciata partially explains the species ability for rapid growth and high productivity.

Use of canonical variates in genetic divergence studies

Exact correlation coefficients between a canonical variate and measured traits were derived to evaluate genetic divergence among varieties. This method allows the plant breeder to determine which traits contribute significantly to genetic divergence and, also, to identify the most important among them. An example is presented, related to a trial where 28 varieties of maize were evaluated for two traits

Use of a cysteine proteinase from Carica candamarcensis as a protective agent during DNA extraction

We describe the use of a plant cysteine proteinase isolated from latex of Carica candamarcensis as a protective agent during isolation of bacterial DNA following growth in culture of these cells. Between 100 to 720 units of proteinase (1 µg = 6 units) afforded good DNA protection when incubated with various kinds of microorganisms. Agarose gel electrophoresis showed that the resulting DNA was similar in size to DNA preparations obtained by treatment with proteinase K. The viability of the resulting material was checked by PCR amplification using species-specific primers. After standing at room temperature (25oC) for 35 days, the enzyme lost 10% of its initial activity. The enzyme stability and good yield of DNA suggest the use of this proteinase as an alternative to proteinase K.

A chromatographic method for the production of a human immunoglobulin G solution for intravenous use

Immunoglobulin G (IgG) of excellent quality for intravenous use was obtained from the cryosupernatant of human plasma by a chromatographic method based on a mixture of ion-exchange, DEAE-Sepharose FF and arginine Sepharose 4B affinity chromatography and a final purification step by Sephacryl S-300 HR gel filtration. The yield of 10 experimental batches produced was 3.5 g IgG per liter of plasma. A solvent/detergent combination of 1% Tri (n-butyl) phosphate and 1% Triton X-100 was used to inactivate lipid-coated viruses. Analysis of the final product (5% liquid IgG) based on the mean for 10 batches showed 94% monomers, 5.5% dimers and 0.5% polymers and aggregates. Anticomplementary activity was 0.3 CH50/mg IgG and prekallikrein activator levels were less than 5 IU/ml. Stability at 37ºC for 30 days in the liquid state was satisfactory. IgG was stored in flasks (2.5 g/flask) at 4 to 8ºC. All the characteristics of the product were consistent with the requirements of the 1997 Pharmacopée Européenne.