Biotechnological Potential of the Stress Response and Plant Cell Death Regulators Proteins in the Biofuel Industry.

Production of biofuel from lignocellulosic biomass is relatively low due to the limited knowledge about natural cell wall loosening and cellulolytic processes in plants. Industrial separation of cellulose fiber mass from lignin, its saccharification and alcoholic fermentation is still cost-ineffective and environmentally unfriendly. Assuming that the green transformation is inevitable and that new sources of raw materials for biofuels are needed, we decided to study cell death-a natural process occurring in plants in the context of reducing the recalcitrance of lignocellulose for the production of second-generation bioethanol. "Members of the enzyme families responsible for lysigenous aerenchyma formation were identified during the root hypoxia stress in Arabidopsis thaliana cell death mutants. The cell death regulatory genes, LESION SIMULATING DISEASE 1 (LSD1), PHYTOALEXIN DEFICIENT 4 (PAD4) and ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1) conditionally regulate the cell wall when suppressed in transgenic aspen. During four years of growth in the field, the following effects were observed: lignin content was reduced, the cellulose fiber polymerization degree increased and the growth itself was unaffected. The wood of transgenic trees was more efficient as a substrate for saccharification, alcoholic fermentation and bioethanol production. The presented results may trigger the development of novel biotechnologies in the biofuel industry.

Influence of Thermal Modification in Nitrogen Atmosphere on the Selected Mechanical Properties of Black Poplar Wood (Populus nigra L.).

The thermal modification of wood in a nitrogen atmosphere shapes the functional properties of wood. The aim of this research was to determine the influence of different levels of temperature and the duration of thermal modification on the mechanical properties of the black poplar (Populus nigra L.). Black poplar was thermally modified in nitrogen atmosphere in the temperature range from 160 °C to 220 °C (6 levels) for 2 h to 8 h (4 levels), resulting in a total of 24 treatments. The effect of these treatments on compressive strength parallel to the grain (CS), modulus of rupture (MOR), and modulus of elasticity during bending (MOE) were analyzed. Thermal modification influenced the mechanical properties of black poplar wood. After thermal modification occurred in a nitrogen atmosphere, an increase in compressive strength was noticeable for all variants of black poplar wood modification. The highest 16% increase in CS was obtained for the modification carried out at the temperature of 160 °C and for 2 h. An increase was also found for MOE when modified under mild conditions, while a decrease occurred for variants at higher temperatures, i.e., for 200 °C and 220 °C. The study showed that for all modification variants, there was a decrease in MOR alongside the increase in modification temperature and time.

Evaluation of the Hydrolysis Efficiency of Bacterial Cellulose Gel Film after the Liquid Hot Water and Steam Explosion Pretreatments.

The influence of bacterial cellulose gel film pretreatment methods on the efficiency of enzymatic hydrolysis was investigated. An increase in the efficiency of enzymatic hydrolysis due to liquid hot water pretreatment or steam explosion was shown. The glucose yield of 88% was obtained from raw, non-purified, bacterial cellulose treated at 130 °C. The results confirm the potential of bacterial cellulose gel film as a source for liquid biofuel production.

Temperature, Time, and Interactions between Them in Relation to Colour Parameters of Black Poplar (Populus nigra L.) Thermally Modified in Nitrogen Atmosphere.

Thermal modification of wood in nitrogen atmosphere permits its usability value to be improved. The aim of the research was to determine the impact of technological modification parameters at different levels on the colour of black poplar (Populus nigra L.). Black poplar was thermally modified in nitrogen atmosphere at a range of temperatures from 160 °C to 220 °C, at times between 2 h and 8 h. The parameters of wood colour were measured according to the CIE L*a*b* colour space model. The changes in a* and b* had a non-linear profile. The maximum value of a* for black poplar wood was achieved after a modification at the temperature of 200 °C, while the maximum value of the b* parameter was achieved after modification at 190 °C. Colour changes in the ΔE of black poplar after modification at 160 °C and 170 °C were similar, and the dynamics of changes increased after modification at the temperature of 180 °C. The highest value of ΔE, around 40, was observed after modification at the temperature of 220 °C and time of 8 h. There were no statistically significant differences between the ΔE for radial and tangential sections. Statistical analysis showed that modification temperature was responsible for the variability of the L* value in 90%, and in ca. 70% for the changes in parameters a* and b*. The influence of the modification time on the colour parameters was minor-below 4%. The influence of the interaction between modification temperature and time on the colour parameters was below 10%. As a consequence, in the case of ΔE of black poplar wood, the influence of temperature was at a level of ca. 80%. On the other hand, the influence of time and the interaction between temperature and time of modification was low-below 3%.

Evaluation of the Dimensional Stability of Black Poplar Wood Modified Thermally in Nitrogen Atmosphere.

Black poplar (Populus nigra L.) was thermally modified in nitrogen atmosphere. The effects of the modification process on poplar wood were evaluated for temperatures: 160 °C, 190 °C, and 220 °C applied for 2 h; and 160 °C and 190 °C for 6 h. The percentual impact of temperature and time of modification on the properties of modified wood was analysed. The study permitted the identification correlations between the chemical composition and selected physical properties of thermally modified poplar wood. The dimensional stability of poplar wood was improved after thermal modification in nitrogen. The higher the temperature of modification, the lower the equilibrium moisture content (EMC) of black poplar. At the temperature of 220 °C, EMC was two times lower than the EMC of non-modified black poplar. It is also possible to reduce the dimensional changes of wood two-fold (at the modification temperature of 220 °C), both in radial and tangential directions, independently of the acclimatization conditions (from 34% to 98% relative humidity, RH). Similar correlations have been found for wood that has been soaked in water. Higher modification temperatures and longer processing times contributed to a lower swelling anisotropy (SA).

PHYTOALEXIN DEFICIENT 4 affects reactive oxygen species metabolism, cell wall and wood properties in hybrid aspen (Populus tremula L. × tremuloides).

The phytoalexin deficient 4 (PAD4) gene in Arabidopsis thaliana (AtPAD4) is involved in the regulation of plant--pathogen interactions. The role of PAD4 in woody plants is not known; therefore, we characterized its function in hybrid aspen and its role in reactive oxygen species (ROS)-dependent signalling and wood development. Three independent transgenic lines with different suppression levels of poplar PAD expression were generated. All these lines displayed deregulated ROS metabolism, which was manifested by an increased H2O2 level in the leaves and shoots, and higher activities of manganese superoxide dismutase (MnSOD) and catalase (CAT) in the leaves in comparison to the wild-type plants. However, no changes in non-photochemical quenching (NPQ) between the transgenic lines and wild type were observed in the leaves. Moreover, changes in the ROS metabolism in the pad4 transgenic lines positively correlated with wood formation. A higher rate of cell division, decreased tracheid average size and numbers, and increased cell wall thickness were observed. The results presented here suggest that the Populus tremula × tremuloides PAD gene might be involved in the regulation of cellular ROS homeostasis and in the cell division--cell death balance that is associated with wood development.