Development and chemical characterization of Casparian strips in the roots of Chinese fir (Cunninghamia lanceolata)

Lignification and suberization of Casparian strips occurred simultaneously during the development of Cunninghamia lanceolata primary roots. The Casparian strips in the cell walls of vascular plant root endodermis and exodermis cells are vital for the exclusion of salts and pathogens, selective nutrient uptake, and other processes related to the transport and assimilates of water. Despite the importance of Casparian strips, their chemical composition and its relevance to their diffusion barrier functions remain a matter of debate, especially in woody plants. Here, we describe the cytological features revealed by fluorescent staining of developing Casparian strips in the primary root endodermis of Chinese fir ( Cunninghamia lanceolata ), one of the most important timber species native to China. Using apoplastic permeability tests, we revealed that the Casparian strips provide their barrier function from the secondary stage of endodermis development, after lignin and suberin are deposited onto the surface of the protoplast next to tangential cell walls. Furthermore, we enzymatically isolated Casparian strips from the primary root endodermis and analyzed their chemical composition using Fourier transform infrared spectroscopy, revealing that they consist largely of lignin and suberin. Using confocal Raman microscopy for label-free, in situ, structural and biochemical analysis, we showed that lignin and suberin accumulated simultaneously in the developing Casparian strips. Taken together, these results elucidated Casparian strip development and characterized the lignification and suberization processes in situ in the primary roots of Chinese fir. Since this species shows broad adaptability to varying environmental conditions, our findings may facilitate work to understand the barrier function of Casparian strips in relationship to environmental stresses.