A technology developed by Christophe Biot and Cédric Lion (Unit of Structural and Fonctional Glycobiology (UGSF), UMR 8576, France)
MonoliF is a labeling kit designed to study lignin biosynthesis inside plants. This kit uses monolignol reporters that are incorporated into lignin just like naturally occuring monolignols. G and S MonoliF can then be detected via fluorescence after labeling with click chemistry. It is the only method that will allow you to study lignin composition and localisation in a dynamic manner, since only the MonoliF (and not naturally occuring monolignols) will be labeled.
Main features of MonoliF
Biological relevance
Modifications on S and G MonoliF cannot be detected by living cell, meaning they are seen as naturally occuring monolignols by plants.
Deeper insights on lignin
You can analyze which type of monolignol is integrated, its localisation, as well as its dynamics.
Live imaging
Lignin dynamics can be studied in seedlings, live stem or cross section of plants (using confocal microscope or airyscan)
How does MonoliF kit work ?
- S and/or G MonoliF analogs bearing either an azide or an alkyne group are metabolically incorporated into lignifying cell walls.
- Detection of MonoliF is performed with click chemistry kit in a straightforward, bioorthogonal manner (the reaction does not interact with natural biochemical processes).
- Lignin incorporation and composition can be studied with fluorescence signal.
Kit content of MonoliF
*One experiment corresponds to 300µL.
Applications
Type of sample : MonoliF has been already validated on cross sections, stems, and seedlings.
Type of plants MonoliF has been tested on Arabidospis Thaliana, Poplar, Tobacco and flax.
Specifications of MonoliF
Fluorescence and microscopy :
- Tagged G-MonoliF : Abs/Em = 546/565 nm
- Tagged S-MonoliF : Abs/Em = 501/526 nm
We recommand using a confocal microscope or an airyscan
Storage and stability :
- Store G and S monoliF at -20°C. Stable up to 4 years
How to use MonoliF ?
Below is a quick protocol overview :
Follow of cell-specific monolignol reporter incorporation with MonoliF (1)
Observation of monolignol incorporation (H monolignol in green, G monolignol in red) in a cross section of flax stem. The incorporation of monolignol varies across the distance from the cambial zone but also according to the type of cell (Ray cells, R, pink arrow; Fiber tracheid cell, FT, yellow arrow)
Versatility of MonoliF labeling across multiple plant types (2)
Incorporation of H (green), G (red), and S (magenta) monolignols in cross-section of Arabidopsis thaliana floral hemp (a), tobacco stem (b) and poplar stem (c).
Labeling of G and S monolignols in flax stem cross section with MonoliF (2)
Identification of lignin (autofluorescence, blue), G monolignol (red) and S monolignol (magenta) in transversal cross-sections (a), longitudinal sections (b) and entire stem (c) of live flax. Scale bar=50 mm (a,b), 10 mm (c) and 5 mm (insert).
Analysis of flax xylem pit border region with MonoliF
Airyscan images of the 3 monolignols reporters : H (in blue), G (in red) and S (in red) BLISS monolignols (A). The merged image shows cell walls and a bordered pit (BP) between two cells in flax xylem (B). Orange lines P1−P3 show vector lines used to trace individual reporter (H, G, S) fluorescence intensity profiles along the lines plotted in (C). Scale bar = 3 μm.
References :
1. Lion et al., Cell Chemical Biology, 2017 http://dx.doi.org/10.1016/j.chembiol.2017.02.009
2. Simon et al., Angew. Chem. Int. Ed. 2018. https://doi.org/10.1002/anie.201808493.
3. Simon & al 2023, Chem Biomed Imaging, https://doi.org/10.1021/cbmi.3c00052
Check out the Publications section for additional results.