Economic potential of essential oil production from New Zealand-grown Eucalyptus bosistoana.

Farm foresters and other growers are establishing a ground-durable hardwood resource, including the emerging plantation species Eucalyptus bosistoana in New Zealand. The foliage of this species contains essential oils in quantity and quality suitable for commercial extraction. Essential oil production could improve the economic viability of E. bosistoana plantations, diversifying the grower's income and providing an early revenue stream. This study assessed the economic potential for essential oil production from New Zealand grown E. bosistoana plantations. A sensitivity analysis indicated that uncertainty of leaf biomass availability, genetic as well as seasonal changes in oil content, and fluctuations in essential oil price are equally important on the viability of an essential oil operation. Small-scale essential oil production could be sustainably supplied with foliage from thinning and pruning operations sourced from the envisaged regional planting programmes and commence in 3-5 years. A large-scale operation could be supplied when trees will be harvested. Lastly, based on the operational costs of a domestic small-scale essential oil producer, oil value from E. bosistoana would exceed the cost of production.

Nanostructural deformation of high-stiffness spruce wood under tension.

Conifer wood is an exceptionally stiff and strong material when its cellulose microfibrils are well aligned. However, it is not well understood how the polymer components cellulose, hemicelluloses and lignin co-operate to resist tensile stress in wood. From X-ray scattering, neutron scattering and spectroscopic data, collected under tension and processed by novel methods, the ordered, disordered and hemicellulose-coated cellulose components comprising each microfibril were shown to stretch together and demonstrated concerted, viscous stress relaxation facilitated by water. Different cellulose microfibrils did not all stretch to the same degree. Attempts were made to distinguish between microfibrils showing large and small elongation but these domains were shown to be similar with respect to orientation, crystalline disorder, hydration and the presence of bound xylan. These observations are consistent with a major stress transfer process between microfibrils being shear at interfaces in direct, hydrogen-bonded contact, as demonstrated by small-angle neutron scattering. If stress were transmitted between microfibrils by bridging hemicelluloses these might have been expected to show divergent stretching and relaxation behaviour, which was not observed. However lignin and hemicellulosic glucomannans may contribute to stress transfer on a larger length scale between microfibril bundles (macrofibrils).

Temperature-Dependent Blue Shifting of O-H Stretching Frequencies in Crystalline Cellulose Explained.

Increasing the temperature of a chemical system generally causes covalent bonds to lengthen and weaken, often the first step in initiating chemical reactions. However, for some hydrogen-bonded systems, infrared (IR) spectroscopy measurements reveal that covalent O-H bonds actually strengthen and therefore shorten when heated. In 1957, Finch and Lippincott proposed a simple one-dimensional (1D) model to explain this effect, in which thermal excitation of intermolecular stretching modes leads to lengthening and weakening of intermolecular O-H···O hydrogen bonds, thereby indirectly strengthening the associated covalent O-H bonds. Taking cellulose (an infinitely repeating polymer of d-glucose) as an example, we use molecular dynamics modeling to show that the same mechanism is responsible for temperature-dependent blue shifting of O-H stretching bands in IR spectra of carbohydrate biopolymers, except that interchain hydrogen bonds are weakened by thermal excitation of chain-separation modes, while intrachain hydrogen bonds are weakened by thermally induced changes in ring puckering and orientation of ring substituents but not reorientation of glucose units relative to one another or overall twisting of the cellulose chains.

Measuring Molecular Strain in Rewetted and Never-Dried Eucalypt Wood with Raman Spectroscopy.

To measure growth strain in wood using Raman spectroscopy, we investigated the Raman spectra of rewetted (water-saturated) Eucalyptus regnans and green Eucalyptus quadrangulata wood during tensile tests. Partial least squares models to predict the tensile strain were built from the Raman spectra. The best model could predict the tensile strain with a root mean square error of 427.5 µÎµ. Apart from the widely reported band shift at 1095 cm-1 upon mechanical strain, spectral changes at 1420, 1120, 895, and 456 cm-1 were identified. The assignments of these bands were discussed in relation to the molecular deformation of cellulose. The band shift rates during tensile tests were -3.06 and -2.15 cm-1/% for rewetted E. regnans and green E. quadrangulata wood, respectively. We successfully detected the release of the molecular growth strain in green eucalyptus wood with Raman spectroscopy by observing band shifts of the 1095 cm-1 signal. Further, there was a moderate correlation (r = 0.48) between the growth strain measured with strain gauges and the 1095 cm-1 band position. The precision of the prediction of growth strain using Raman spectroscopy was negatively affected by variation attributed to the inhomogeneity of wood on the millimeter scale and instrumental instability.

Effects of variable selection and processing of NIR and ATR-IR spectra on the prediction of extractive content in Eucalyptus bosistoana heartwood.

The use of quick attenuated total reflectance infrared (ATR-IR) spectroscopy and near infrared (NIR) spectroscopy to predict extractives content (EC) in heartwood of E. bosistoana with partial least squares regression (PLSR) models was studied. Different spectra pre-processing methods and variable selection were tested for calibration optimisation. While variable selection substantially improved the NIR-PLSR models, only small effects were observed for spectra pre-processing methods and ATR-IR-PLSR models. Both of the NIR-PLSR and ATR-IR-PLSR models yielded reliably EC results with high R2 and low root mean square error (RMSE). NIR based models performed better (RMSE 0.9%) than ATR-IR based models (RMSE 1.6%). Analysis showed that the models were based on IR signals assigned to chemical structures known from eucalyptus heartwood extracts. Combined with PLSR and variable selection, both, ATR-IR and the NIR spectroscopy, can be used to quickly predict EC in E. bosistoana, a measure needed in tree breeding and the quality control of for durable timber.

Effects of mechanical stretching, desorption and isotope exchange on deuterated eucalypt wood studied by near infrared spectroscopy.

Deuterium exchange combined with near infrared (NIR) spectroscopy was used to study the roles of accessible and inaccessible cellulose in the load transfer of eucalyptus wood. Monitoring the drying process helped to assign NIR bands of deuterated wood samples. Polarized NIR spectra of protonated and deuterated samples confirmed that inaccessible hydroxyl groups in eucalyptus wood were preferably oriented in the longitudinal direction. The spectral changes on NIR spectra caused by mechanical strain could be highlighted by averaging loading and unloading cycles to compensate for effects of desorption and isotope re-exchange due to environmental fluctuations. After deuteration, the bands affected by mechanical strain at around 6420, 6240 and 4670 cm-1, which had been assigned to hydroxyl groups in cellulose, remained at these positions, suggesting the inaccessible cellulose fraction was the main load-bearing component in wood. A small band at around 4700 cm-1 responding to mechanical strain, becoming visible in the deuterated spectra, indicated that accessible hydroxyls also contributed to the load transfer. Furthermore, the measurements confirmed previous reports of moisture adsorption of wood under tensile stress.

Molecular deformation of wood and cellulose studied by near infrared spectroscopy.

Wood (Eucalyptus regnans and Pinus radiata) and paper samples were stretched to different strain levels using a purpose-built tensile test device fitted into a near infrared (NIR) spectrometer while collecting transmission spectra. Consistent spectral changes caused by mechanical strain, assigned to OH stretching bands, were observed for all three sample types. Bands at 6286 ±â€¯5 cm-1 and 6470 ±â€¯10 cm-1 were tentatively assigned to the OH groups connected with the 2OH⋯6O and 3OH⋯5O intramolecular hydrogen bonds of crystalline cellulose Iß, respectively. Both bands shifted to higher wavenumbers indicating the elongation of the hydrogen bonds. A linear relationship was found between band shifts and mechanical strain. Band shift rates for the 3OH bond were more than twice that of the 2OH bond, consistent with bending of the glycosidic bond. Bending tests showed that the band at around 6286 cm-1 shifted in opposite direction when under tension or compression.

Predicting extractives content of Eucalyptus bosistoana F. Muell. Heartwood from stem cores by near infrared spectroscopy.

Time and resource are the restricting factors for the wider use of chemical information of wood in tree breeding programs. NIR offers an advantage over wet-chemical analysis in these aspects and is starting to be used for tree breeding. This work describes the development of a NIR-based assessment of extractive content in heartwood of E. bosistoana, which does not require milling and conditioning of the samples. This was achieved by applying the signal processing algorithms (external parameter orthogonalisation (EPO) and significance multivariate correlation (sMC)) to spectra obtained from solid wood cores, which were able to correct for moisture content, grain direction and sample form. The accuracy of extractive content predictions was further improved by variable selection, resulting in a root mean square error of 1.27%. Considering the range of extractive content in E. bosistoana heartwood of 1.3 to 15.0%, the developed NIR calibration has the potential to be used in an E. bosistoana breeding program or to assess the special variation in extractive content throughout a stem.

Diffraction evidence for the structure of cellulose microfibrils in bamboo, a model for grass and cereal celluloses.

BACKGROUND: Cellulose from grasses and cereals makes up much of the potential raw material for biofuel production. It is not clear if cellulose microfibrils from grasses and cereals differ in structure from those of other plants. The structures of the highly oriented cellulose microfibrils in the cell walls of the internodes of the bamboo Pseudosasa amabilis are reported. Strong orientation facilitated the use of a range of scattering techniques. RESULTS: Small-angle neutron scattering provided evidence of extensive aggregation by hydrogen bonding through the hydrophilic edges of the sheets of chains. The microfibrils had a mean centre-to-centre distance of 3.0 nm in the dry state, expanding on hydration. The expansion on hydration suggests that this distance between centres was through the hydrophilic faces of adjacent microfibrils. However in the other direction, perpendicular to the sheets of chains, the mean, disorder-corrected Scherrer dimension from wide-angle X-ray scattering was 3.8 nm. It is possible that this dimension is increased by twinning (crystallographic coalescence) of thinner microfibrils over part of their length, through the hydrophobic faces. The wide-angle scattering data also showed that the microfibrils had a relatively large intersheet d-spacing and small monoclinic angle, features normally considered characteristic of primary-wall cellulose. CONCLUSIONS: Bamboo microfibrils have features found in both primary-wall and secondary-wall cellulose, but are crystallographically coalescent to a greater extent than is common in celluloses from other plants. The extensive aggregation and local coalescence of the microfibrils are likely to have parallels in other grass and cereal species and to influence the accessibility of cellulose to degradative enzymes during conversion to liquid biofuels.

How cellulose stretches: synergism between covalent and hydrogen bonding.

Cellulose is the most familiar and most abundant strong biopolymer, but the reasons for its outstanding mechanical performance are not well understood. Each glucose unit in a cellulose chain is joined to the next by a covalent C-O-C linkage flanked by two hydrogen bonds. This geometry suggests some form of cooperativity between covalent and hydrogen bonding. Using infrared spectroscopy and X-ray diffraction, we show that mechanical tension straightens out the zigzag conformation of the cellulose chain, with each glucose unit pivoting around a fulcrum at either end. Straightening the chain leads to a small increase in its length and is resisted by one of the flanking hydrogen bonds. This constitutes a simple form of molecular leverage with the covalent structure providing the fulcrum and gives the hydrogen bond an unexpectedly amplified effect on the tensile stiffness of the chain. The principle of molecular leverage can be directly applied to certain other carbohydrate polymers, including the animal polysaccharide chitin. Related but more complex effects are possible in some proteins and nucleic acids. The stiffening of cellulose by this mechanism is, however, in complete contrast to the way in which hydrogen bonding provides toughness combined with extensibility in protein materials like spider silk.

Structure of cellulose microfibrils in primary cell walls from collenchyma.

In the primary walls of growing plant cells, the glucose polymer cellulose is assembled into long microfibrils a few nanometers in diameter. The rigidity and orientation of these microfibrils control cell expansion; therefore, cellulose synthesis is a key factor in the growth and morphogenesis of plants. Celery (Apium graveolens) collenchyma is a useful model system for the study of primary wall microfibril structure because its microfibrils are oriented with unusual uniformity, facilitating spectroscopic and diffraction experiments. Using a combination of x-ray and neutron scattering methods with vibrational and nuclear magnetic resonance spectroscopy, we show that celery collenchyma microfibrils were 2.9 to 3.0 nm in mean diameter, with a most probable structure containing 24 chains in cross section, arranged in eight hydrogen-bonded sheets of three chains, with extensive disorder in lateral packing, conformation, and hydrogen bonding. A similar 18-chain structure, and 24-chain structures of different shape, fitted the data less well. Conformational disorder was largely restricted to the surface chains, but disorder in chain packing was not. That is, in position and orientation, the surface chains conformed to the disordered lattice constituting the core of each microfibril. There was evidence that adjacent microfibrils were noncovalently aggregated together over part of their length, suggesting that the need to disrupt these aggregates might be a constraining factor in growth and in the hydrolysis of cellulose for biofuel production.

An unusual form of reaction wood in Koromiko [Hebe salicifolia G. Forst. (Pennell)], a southern hemisphere angiosperm.

Koromiko [Hebe salicifolia G. Forst. (Pennell)] is a woody angiosperm native to New Zealand and Chile. Hebe spp. belong to the otherwise herbaceous family Plantaginaceae in the order Lamiales. Reaction wood exerting expansional forces was found on the lower side of leaning H. salicifolia stems. Such reaction wood is atypical for angiosperms, which commonly form contracting reaction wood on the upper side of leaning stems. Reaction wood typical for angiosperms is formed by species in other families in the order Lamiales. This suggests that the form of reaction wood is specific to the family level. Functionally the reaction wood of H. salicifolia is similar to that found in gymnosperms, which both act by pushing. However, their chemical, anatomical and physical characteristics are different. Typical features of reaction wood present in gymnosperms such as high density, thick-walled rounded cells and the presence of (1 → 4)-ß-galactan in the secondary cell wall layer are absent in H. salicifolia reaction wood. Reaction wood of H. salicifolia varies from normal wood in having a higher microfibril angle, which is likely to determine the direction of generated maturation stresses.

Nanostructure of cellulose microfibrils in spruce wood.

The structure of cellulose microfibrils in wood is not known in detail, despite the abundance of cellulose in woody biomass and its importance for biology, energy, and engineering. The structure of the microfibrils of spruce wood cellulose was investigated using a range of spectroscopic methods coupled to small-angle neutron and wide-angle X-ray scattering. The scattering data were consistent with 24-chain microfibrils and favored a "rectangular" model with both hydrophobic and hydrophilic surfaces exposed. Disorder in chain packing and hydrogen bonding was shown to increase outwards from the microfibril center. The extent of disorder blurred the distinction between the I alpha and I beta allomorphs. Chains at the surface were distinct in conformation, with high levels of conformational disorder at C-6, less intramolecular hydrogen bonding and more outward-directed hydrogen bonding. Axial disorder could be explained in terms of twisting of the microfibrils, with implications for their biosynthesis.

Distribution of (1->4)-beta-galactans, arabinogalactan proteins, xylans and (1->3)-beta-glucans in tracheid cell walls of softwoods.

Polysaccharides were located in the walls of normal and compression wood tracheids of Pinus radiata (radiata pine), Picea sitchensis (Sitka spruce) and Picea abies (Norway spruce) by transmission electron microscopy using immunogold labelling with monoclonal antibodies to (1-->4)-beta-galactan (LM5), (1-->3)-beta-glucan, arabinogalactan proteins (AGPs) (MAC207) and heteroxylans (LM10 and LM11). In fully differentiated compression wood tracheids, (1-->4)-beta-galactan was found in the S2((L)) layer and, to a smaller extent, at the interface between the compound middle lamella and the S1 layer. (1-->4)-beta-Galactan appeared to be displaced from, or modified in, the S1 layer during cell wall formation. (1-->3)-beta-Glucan (callose) was confined to the helical cavities in the inner S2 layer of severe compression wood. MAC207 AGP glycan epitope was found exclusively in the S1 and S3 layers of normal wood tracheids and in the S1 and inner S2 layers of compression wood tracheids. Binding of LM10, which specifically recognizes unsubstituted or low-substituted xylans, occurred at similar locations to the MAC207 epitope, whereas binding of LM11, which recognizes more highly substituted as well as unsubstituted xylans, occurred throughout the tracheid walls with the exception of the primary wall. Immunogold labelling showed that the different wall layers of softwood tracheids have different polysaccharide compositions which change abruptly during cell wall formation.

Regioselective deacetylation of cellulose acetates by acetyl xylan esterases of different CE-families.

Cellulose acetate (CA) was found to be a substrate of several acetyl xylan esterases (AXE). Eight AXE from different carbohydrate esterase (CE) families were tested on their activity against CA with a degree of substitution of 0.7 and 1.4. The classification of the AXEs into CE families according to their structure by hydrophobic cluster analysis followed clearly their activity against CA. Within the same CE family similar, and between the CE families different deacetylation behaviours could be observed. Furthermore, each esterase family showed a distinct regioselective mode of action. The CE 1 family enzymes regioselectively cleaved the substituents in C2- and C3-position, while CE 5 family enzymes only cleaved the acetyl groups in C2-position. CE 4 family enzymes seemed to interact only with the substituents in C3-position. Evidence was found that the deacetylation reaction of the CE 1 family enzymes proceeded faster in C2- than in C3-position of CA. The enzymes were able to cleave acetyl groups from fully substituted anhydroglucose units.