Report on the Current Inventory of the Toolbox for Plant Cell Wall Analysis: Proteinaceous and Small Molecular Probes.

Plant cell walls are highly complex structures composed of diverse classes of polysaccharides, proteoglycans, and polyphenolics, which have numerous roles throughout the life of a plant. Significant research efforts aim to understand the biology of this cellular organelle and to facilitate cell-wall-based industrial applications. To accomplish this, researchers need to be provided with a variety of sensitive and specific detection methods for separate cell wall components, and their various molecular characteristics in vitro as well as in situ. Cell wall component-directed molecular detection probes (in short: cell wall probes, CWPs) are an essential asset to the plant glycobiology toolbox. To date, a relatively large set of CWPs has been produced-mainly consisting of monoclonal antibodies, carbohydrate-binding modules, synthetic antibodies produced by phage display, and small molecular probes. In this review, we summarize the state-of-the-art knowledge about these CWPs; their classification and their advantages and disadvantages in different applications. In particular, we elaborate on the recent advances in non-conventional approaches to the generation of novel CWPs, and identify the remaining gaps in terms of target recognition. This report also highlights the addition of new "compartments" to the probing toolbox, which is filled with novel chemical biology tools, such as metabolic labeling reagents and oligosaccharide conjugates. In the end, we also forecast future developments in this dynamic field.

Surface properties correlate to the digestibility of hydrothermally pretreated lignocellulosic Poaceae biomass feedstocks.

BACKGROUND: Understanding factors that govern lignocellulosic biomass recalcitrance is a prerequisite for designing efficient 2nd generation biorefining processes. However, the reasons and mechanisms responsible for quantitative differences in enzymatic digestibility of various biomass feedstocks in response to hydrothermal pretreatment at different severities are still not sufficiently understood. RESULTS: Potentially important lignocellulosic feedstocks for biorefining, corn stover (Zea mays subsp. mays L.), stalks of Miscanthus × giganteus, and wheat straw (Triticum aestivum L.) were systematically hydrothermally pretreated; each at three different severities of 3.65, 3.83, and 3.97, respectively, and the enzymatic digestibility was assessed. Pretreated samples of Miscanthus × giganteus stalks were the least digestible among the biomass feedstocks producing ~24 to 66.6% lower glucose yields than the other feedstocks depending on pretreatment severity and enzyme dosage. Bulk biomass composition analyses, 2D nuclear magnetic resonance, and comprehensive microarray polymer profiling were not able to explain the observed differences in recalcitrance among the pretreated feedstocks. However, methods characterizing physical and chemical features of the biomass surfaces, specifically contact angle measurements (wettability) and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy (surface biopolymer composition) produced data correlating pretreatment severity and enzymatic digestibility, and they also revealed differences that correlated to enzymatic glucose yield responses among the three different biomass types. CONCLUSION: The study revealed that to a large extent, factors related to physico-chemical surface properties, namely surface wettability as assessed by contact angle measurements and surface content of hemicellulose, lignin, and wax as assessed by ATR-FTIR rather than bulk biomass chemical composition correlated to the recalcitrance of the tested biomass types. The data provide new insight into how hydrothermal pretreatment severity affects surface properties of key Poaceae lignocellulosic biomass and may help design new approaches to overcome biomass recalcitrance.

Three Pectin Methylesterase Inhibitors Protect Cell Wall Integrity for Arabidopsis Immunity to Botrytis.

Infection by necrotrophs is a complex process that starts with the breakdown of the cell wall (CW) matrix initiated by CW-degrading enzymes and results in an extensive tissue maceration. Plants exploit induced defense mechanisms based on biochemical modification of the CW components to protect themselves from enzymatic degradation. The pectin matrix is the main CW target of Botrytis cinerea, and pectin methylesterification status is strongly altered in response to infection. The methylesterification of pectin is controlled mainly by pectin methylesterases (PMEs), whose activity is posttranscriptionally regulated by endogenous protein inhibitors (PMEIs). Here, AtPMEI10, AtPMEI11, and AtPMEI12 are identified as functional PMEIs induced in Arabidopsis (Arabidopsis thaliana) during B. cinerea infection. AtPMEI expression is strictly regulated by jasmonic acid and ethylene signaling, while only AtPMEI11 expression is controlled by PME-related damage-associated molecular patterns, such as oligogalacturonides and methanol. The decrease of pectin methylesterification during infection is higher and the immunity to B. cinerea is compromised in pmei10, pmei11, and pmei12 mutants with respect to the control plants. A higher stimulation of the fungal oxalic acid biosynthetic pathway also can contribute to the higher susceptibility of pmei mutants. The lack of PMEI expression does not affect hemicellulose strengthening, callose deposition, and the synthesis of structural defense proteins, proposed as CW-remodeling mechanisms exploited by Arabidopsis to resist CW degradation upon B. cinerea infection. We show that PME activity and pectin methylesterification are dynamically modulated by PMEIs during B. cinerea infection. Our findings point to AtPMEI10, AtPMEI11, and AtPMEI12 as mediators of CW integrity maintenance in plant immunity.