measuring-interactions-between-fluorescent-probes-lignin-plant

Measuring Interactions between Fluorescent Probes and Lignin in Plant Sections by sFLIM Based on Native Autofluorescence

In lignocellulosic biomass (LB), the activity of enzymes is limited by the appearance of non-specific interactions with lignin during the hydrolysis ...

Université de Reims Champagne-Ardenne

Mid-infrared (MIR) and near-infrared (NIR) spectroscopy provide useful information on the molecular composition of biological systems. Because they are sensitive to organic and mineral components, there is a growing interest in these techniques for the development of biomarkers that reflect intrinsic characteristics of plants and their mode of degradation. Due to their complexity and complementary nature, an important challenge is the combining of MIR and NIR information to identify discriminating wavenumbers in each wavenumber region, with the ultimate goal of assessing the biodegradation process of a lignocellulosic biomass at different time scales. This work investigates the potential of using the outer product to combine MIR and NIR spectra to highlight the connections between fundamental molecular vibrations and their combinations and bonds. Because this operation yields high-dimensional spectra, we propose to use a genetic algorithm to select the most discriminant wavenumbers within the degradation process. The results from two lignocellulosic biomasses with different biodegradation kinetics, miscanthus aerial parts and maize roots, confirm that the outer product combination of MIR and NIR spectral information allows a better discrimination of the biodegradation kinetic compared with the simple concatenation of MIR and NIR spectra or with the use of MIR or MIR spectral information separately. We show that the genetic algorithm selects wavenumbers that correspond to principal vibrations of chemical functional groups of compounds that undergo degradation/conversion during the biodegradation of the lignocellulosic biomass. 

Evaluation of Lignocellulosic Biomass Degradation by Combining Mid- and Near-Infrared Spectra by the Outer Product and Selecting Discriminant Wavenumbers Using a Genetic Algorithm.

The lignocellulosic cell wall network is resistant to enzymatic degradation due to the complex chemical and structural features. Pretreatments are thus commonly used to overcome natural recalcitrance of lignocellulose. Characterization of their impact on architecture requires combinatory approaches. However, the accessibility of the lignocellulosic cell walls still needs further insights to provide relevant information.

Exploring accessibility of pretreated poplar cell walls by measuring dynamics of fluorescent probes.

Interest in hemp (Cannabis sativa L.) is increasing due to the development of a new range of industrial applications based on bast fibers. However the variability of bast fiber yield and quality represents an important barrier to further exploitation. Primary and secondary fiber content was examined in two commercial hemp varieties (Fedora 17, Santhica 27) grown under contrasted sowing density and irrigation conditions. Both growing conditions and hemp varieties impact stem tissue architecture with a large effect on the proportion of secondary fibers but not primary fibers. Attenuated total reflectance infrared spectroscopy allowed the discrimination of manually-isolated native primary fibers and secondary fibers but did not reveal any clustering according to growing conditions and variety. Infrared data were confirmed by wet chemistry analyses that revealed slight but significant differences between primary and secondary fiber cell wall composition. Infrared spectroscopy of technical fibers obtained after mechanical defibering revealed differences with native primary, but not secondary fibers and also discriminated samples obtained from plants grown under different conditions. Altogether the results suggested that the observed variability of hemp technical fibers could be partially explained by i) differences in secondary fiber production and ii) differential behavior during mechanical defibering resulting in unequal separation of primary and secondary fibers.

Changes in hemp secondary fiber production related to technical fiber variability revealed by light microscopy and attenuated total reflectance Fourier transform infrared spectroscopy.

Lignocellulosic biomass is a complex network of polymers making up the cell walls of plants. It represents a feedstock of sustainable resources to be converted into fuels, chemicals, and materials. Because of its complex architecture, lignocellulose is a recalcitrant material that requires some pretreatments and several types of catalysts to be transformed efficiently. Gaining more knowledge in the architecture of plant cell walls is therefore important to understand and optimize transformation processes. For the first time, super-resolution imaging of poplar wood samples has been performed using the Stimulated Emission Depletion (STED) technique. In comparison to standard confocal images, STED reveals new details in cell wall structure, allowing the identification of secondary walls and middle lamella with fine details, while keeping open the possibility to perform topochemistry by the use of relevant fluorescent nano-probes. In particular, the deconvolution of STED images increases the signal-to-noise ratio so that images become very well defined. The obtained results show that the STED super-resolution technique can be easily implemented by using cheap commercial fluorescent rhodamine-PEG nano-probes which outline the architecture of plant cell walls due to their interaction with lignin. Moreover, the sample preparation only requires easily-prepared plant sections of a few tens of micrometers, in addition to an easily-implemented post-treatment of images. Overall, the STED super-resolution technique in combination with a variety of nano-probes can provide a new vision of plant cell wall imaging by filling in the gap between classical photon microscopy and electron microscopy.

Anouck Habrant

Fractionation of AgroResources and Environment (FARE) Laboratory,
INRA,
Fractionation of AgroResources and Environment (FARE) Laboratory, INRA

Université de Reims Champagne-Ardenne

Measuring Interactions between Fluorescent Probes and Lignin in Plant Sections by sFLIM Based on Native Autofluorescence

Anouck Habrant

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Université de Reims Champagne-Ardenne

Measuring Interactions between Fluorescent Probes and Lignin in Plant Sections by sFLIM Based on Native Autofluorescence

Fractionation of AgroResources and Environment (FARE) Laboratory,
INRA,
Fractionation of AgroResources and Environment (FARE) Laboratory, INRA