Biomass formation and sugar release efficiency of Populus modified by altered expression of a NAC transcription factor

Woody biomass is an important feedstock for biofuel production. Manipulation of wood properties that enable efficient conversion of biomass to biofuel reduces cost of biofuel production. Wood cell wall composition is regulated at several levels that involve expression of transcription factors such as wood-/secondary cell wall- associated NAC domains (WND or SND). In Arabidopsis thaliana, SND1 regulates cell wall composition through activation of its down-stream targets such as MYBs. The functional aspects of SND1 homologs in the woody Populus have been studied through transgenic manipulation. In this study, we investigated the role of PdWND1B, Populus SND1 sequence ortholog, in wood formation using transgenic manipulation through over-expression or silencing under the control of a vascular-specific 4-coumarate-CoA ligase (4CL) promoter. As compared to control plants, PdWND1B-RNAi plants were shorter in height, with significantly reduced stem diameter and dry biomass, whereas there were no significant differences in growth and productivity of PdWND1B over-expression plants. Conversely, PdWND1B over-expression lines showed a significant reduction in cellulose and increase in lignin content, whereas there was no significant impact on lignin content of down-regulated lines. Stem carbohydrate composition analysis revealed a decrease in glucose, mannose, arabinose, and galactose, but an increase in xylose in the over-expression lines. Transcriptome analysis revealed upregulation of several downstream transcription factors and secondary cell wall related structural genes in the PdWND1B over-expression lines that corresponded to significant phenotypic changes in cell wall chemistry observed in PdWND1B overexpression lines. Relative to the control, glucose release and ethanol production from stem biomass was significantly reduced in over-expression lines but appeared enhanced in the RNAi lines. Our results show that PdWND1B is an important factor determining biomass productivity, cell wall chemistry and its conversion to biofuels in Populus.

Tailoring renewable materials via plant biotechnology

Plants inherently display a rich diversity in cell wall chemistry, as they synthesize an array of polysaccharides along with lignin, a polyphenolic that can vary dramatically in subunit composition and interunit linkage complexity. These same cell wall chemical constituents play essential roles in our society, having been isolated by a variety of evolving industrial processes and employed in the production of an array of commodity products to which humans are reliant. However, these polymers are inherently synthesized and intricately packaged into complex structures that facilitate plant survival and adaptation to local biogeoclimatic regions and stresses, not for ease of deconstruction and commercial product development. Herein, we describe evolving techniques and strategies for altering the metabolic pathways related to plant cell wall biosynthesis, and highlight the resulting impact on chemistry, architecture, and polymer interactions. Furthermore, this review illustrates how these unique targeted cell wall modifications could significantly extend the number, diversity, and value of products generated in existing and emerging biorefineries. These modifications can further target the ability for processing of engineered wood into advanced high performance materials. In doing so, we attempt to illuminate the complex connection on how polymer chemistry and structure can be tailored to advance renewable material applications, using all the chemical constituents of plant-derived biopolymers, including pectins, hemicelluloses, cellulose, and lignins.

Plant cell wall chemistry: implications for ruminant utilisation

Ruminants have adapted to cope with bulky, fibrous forage diets by accommodating a large, diverse microbial population in the reticulo-rumen. Ruminants are dependent on forages as their main sources of energy and other nutrients. Forages are comprised of a complex matrix of cellulose, hemicellulose, protein, minerals and phenolic compounds (including lignin and tannins) with various linkages; many of which are poorly defined. The composition and characteristics of polysaccharides vary greatly among forages and plant cell walls. Plant cell walls are linked and packed together in tight configurations to resist degradation, and hence their nutritional value to animals varies considerably, depending on composition, structure and degradability. An understanding of the inter-relationship between the chemical composition and the degradation of plant cell walls by rumen microorganisms is of major economic importance to ruminant production. Increasing the efficiency of fibre degradation in the rumen has been the subject of extensive research for many decades. This review summarises current knowledge of forage chemistry in order to develop strategies to increase efficiency of forage utilisation by ruminants.

Silica formation in sorghum (Sorghum bicolor) roots

<p>Silicon oxides are the most abundant mineral group in soils. Therefore, plant roots are always exposed to some silicic acid (Si(OH)<sub>4</sub>), which is the soluble form of silicates. Monosilicic acid molecules are taken up by roots, carried in the xylem, and subsequently polymerize to silica in varied silicifying target sites. This biogenic silica (SiO<sub>2</sub>·<em>n</em>H<sub>2</sub>O) can constitute several percent by dry weight in certain plant taxa. However, the mechanisms of its formation remain mostly unknown. In the roots of sorghum (<em>Sorghum bicolor</em>), silica aggregates form in an orderly pattern along the cell walls of endodermis cells. To investigate the structure and composition of root silica aggregates, we studied their development along roots of hydroponically grown sorghum seedlings. By using Raman micro-spectroscopy, auto-fluorescence, and scanning electron microscopy, we found that putative silica aggregation loci could be identified in roots grown under Si starvation. These micrometer-scale spots were constructed of tightly packed modified lignin and were capable of nucleating trace concentrations of silicic acid. Substantial variation in cell wall auto-fluorescence between roots grown with and without silicic acid demonstrated the impact of silicon on cell wall chemistry. Taken together, this work demonstrates a high degree of control over lignin and silica deposition in cell walls. Such regulation implies an important, yet unknown, function for silicon in plant biology.</p>

Characterizing Fungal Decay of Beech Wood: Potential for Biotechnological Applications

The biotechnological potential of nine decay fungi collected from stored beech logs at a pulp and paper factory yard in Northern Iran was investigated. Beech blocks exposed to the fungi in a laboratory decay test were used to study changes in cell wall chemistry using both wet chemistry and spectroscopic methods. Pleurotus ostreatus, P. pulmonarius, and Lentinus sajor-caju caused greater lignin breakdown compared to other white-rot fungi, which led to a 28% reduction in refining energy. Trametesversicolor caused the greatest glucan loss, while P. ostreatus and L. sajor-caju were associated with the lowest losses of this sugar. Fourier transform infrared spectroscopy (FTIR) analyses indicated that white-rot fungi caused greater lignin degradation in the cell walls via the oxidation aromatic rings, confirming the chemical analysis. The rate of cellulose and lignin degradation by the T.versicolor and Pleurotus species was high compared to the other decay fungi analyzed in this study. Based on the above information, we propose that, among the fungi tested, P. ostreatus (27.42% lignin loss and 1.58% cellulose loss) and L. sajor-caju (29.92% lignin loss and 5.95% cellulose loss) have the greatest potential for biopulping.

Winter Nights during Summer Time: Stress Physiological Response to Ice and the Facilitation of Freezing Cytorrhysis by Elastic Cell Wall Components in the Leaves of a Nival Species

Ranunculus glacialis grows and reproduces successfully, although the snow-free time period is short (2–3 months) and night frosts are frequent. At a nival site (3185 m a.s.l.), we disentangled the interplay between the atmospheric temperature, leaf temperatures, and leaf freezing frequency to assess the actual strain. For a comprehensive understanding, the freezing behavior from the whole plant to the leaf and cellular level and its physiological after-effects as well as cell wall chemistry were studied. The atmospheric temperatures did not mirror the leaf temperatures, which could be 9.3 °C lower. Leaf freezing occurred even when the air temperature was above 0 °C. Ice nucleation at on average −2.6 °C started usually independently in each leaf, as the shoot is deep-seated in unfrozen soil. All the mesophyll cells were subjected to freezing cytorrhysis. Huge ice masses formed in the intercellular spaces of the spongy parenchyma. After thawing, photosynthesis was unaffected regardless of whether ice had formed. The cell walls were pectin-rich and triglycerides occurred, particularly in the spongy parenchyma. At high elevations, atmospheric temperatures fail to predict plant freezing. Shoot burial prevents ice spreading, specific tissue architecture enables ice management, and the flexibility of cell walls allows recurrent freezing cytorrhysis. The peculiar patterning of triglycerides close to ice rewards further investigation.

Using structural biology of methanogen enzymes to aid our understanding of methane formation

Methane is a potent greenhouse gas (28-fold more potent than carbon dioxide) and is a significant gas contributing to global climate change. Approximately a billion tons of methane are produced each year by methanogenic archaea in ruminants. These archaea possess a number of unusual traits such as isoprenoid-based lipids, unusual cell wall chemistry and a unique energy metabolism (methanogenesis) that requires six methanogen-specific cofactors. Many of the enzymes involved in these processes have no direct analogues in the host animal. To gain insights into the fundamental biology of rumen methanogens we have determined crystal structures of key enzymes with archaeal-specific traits. Over 600 enzymes were targeted for structure determination which produced approximately 200 purified soluble enzymes for crystallographic screening. More than 50 different enzymes have produced crystals and 30 structures have been solved for individual enzymes to date. The results have helped illuminate our understanding of methane formation at this critical juncture in the world’s history.