Length determination of vessel elements in tree trunks used for water and nutrient transport by Fourier transform Raman spectroscopy.

Length analysis of vessel elements in tree trunks used for water and nutrient transport is a lengthy, multistep procedure although it reflects environmental stresses on a tree. The feasibility of using FT-Raman spectroscopy for rapid determination of vessel element length in a tree was examined using wood powders of two Eucalyptus species, including samples of various ages and colors. The first-derivative transformation followed by the multiplicative scatter correction of Raman spectroscopic data and the partial least-squares regression revealed highly significant correlation between conventionally measured and Raman-predicted vessel element length with correlation coefficients (r) of 0.843 and 0.826, respectively, in the calibration (for known samples, n = 186) and in the prediction (for unknown samples, n = 40). FT-Raman spectroscopy coupled with multivariate data analysis will contribute to solving the interactions between emerging environmental issues and the anatomical structure of wood, which allow efficient management practices in growing forests to fix atmospheric CO(2) effectively.

Sodium blocking induced by a point mutation at the C-terminal end of the pore helix of the KAT1 channel.

A plant hyperpolarization-activating K+ channel, KAT1, is highly selective for K+ over Na+ and is little affected by external Na+, which is crucial to take up K+ effectively in a Na+-containing environment. It has been shown that a mutation at the location (Thr256) preceding the selectivity signature sequence dramatically enhanced the sensitivity of the KAT1 channel to external Na+. We report here electrophysiological experiments for the mechanism of action of external Na+ on KAT1 channels. The Thr256 residue was substituted with either glutamine (Q) or glutamate (E). The wild-type channel was insensitive to external Na+. However, the activity of both mutant channels was significantly depressed by Na+ with apparent dissociation constants of 6.7 mm and 11.3 mm for T256Q and T256E, respectively. The instantaneous current-voltage relationships revealed distinct blocking mechanisms for these mutants. For T256Q a typical voltage-dependent fast blocking was shown. On the other hand, the blocking for the T256E mutant was voltage-independent at low Na+ concentrations and became voltage-dependent at higher concentrations. At extreme hyperpolarization the blocking was relieved significantly. These data strongly suggest that the mutation at the end of the pore helix rearranged the selectivity filter and allows Na+ to penetrate into the pore.

Quantitative analysis of detailed lignin monomer composition by pyrolysis-gas chromatography combined with preliminary acetylation of the samples.

Detailed quantitative analysis of lignin monomer composition comprising p-coumaryl, coniferyl, and sinapyl alcohol and p-coumaraldehyde, coniferaldehyde, and sinapaldehyde in plant has not been studied from every point mainly because of artifact formation during the lignin isolation procedure, partial loss of the lignin components inherent in the chemical degradative methods, and difficulty in the explanation of the complex spectra generally observed for the lignin components. Here we propose a new method to quantify lignin monomer composition in detail by pyrolysis-gas chromatography (Py-GC) using acetylated lignin samples. The lignin acetylation procedure would contribute to prevent secondary formation of cinnamaldehydes from the corresponding alcohol forms during pyrolysis, which are otherwise unavoidable in conventional Py-GC process to some extent. On the basis of the characteristic peaks on the pyrograms of the acetylated sample, lignin monomer compositions in various dehydrogenative polymers (DHP) as lignin model compounds were determined, taking even minor components such as cinnamaldehydes into consideration. The observed compositions by Py-GC were in good agreement with the supplied lignin monomer contents on DHP synthesis. The new Py-GC method combined with sample preacetylation allowed us an accurate quantitative analysis of detailed lignin monomer composition using a microgram order of extractive-free plant samples.

In situ determination of proportion of cell types in wood by Fourier transform Raman spectroscopy.

Analysis of the proportion of cell types in native wood is important for understanding the environmental stresses including an increasing atmospheric CO2 concentration on the structure of wood, especially for the management of plantation forests which will reduce our reliance on natural forests. The conventional method for determining the proportion of cell types is a quantitative microscopy, which is one of the image analyzing systems using a light microscope combined with a microcomputer. However, it is a lengthy multistep procedure. We have examined the feasibility of using Fourier transform Raman spectroscopy for rapid determination of proportion of cell types (fiber, ray parenchyma, vessels, and axial parenchyma) in native wood with using wood meals of two Eucalyptus species, including samples of various ages and colors. By the application of second derivative transformation of Raman spectroscopic data and the partial least-squares regression, we have successfully obtained highly significant correlations between microscopically measured and Raman predicted values for all traits except vessels with correlation coefficients of >0.9 and 0.8, respectively, in the calibration and in the prediction. This method is valid for all traits since vessels can be calculated by the rest of three traits, and will help to solve the effect of the environmental issues on trees and the supplement of renewable raw materials.