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VLCFA-mediated inter-cell layer communication controls cellular pluripotency in Arabidopsis callus

Doll, Y.; Nobusawa, T.; Kojima, M.; Nagata, K.; Matsuda-Ito, K.; Mähönen, A. P.; Matsuda, T.; Abe, M.; Sakakibara, H.; Ikeuchi, M.

2026-09-03 plant biology
10.64898/2026.09.01.748728 bioRxiv
Show abstract

Plants have remarkable capacity to reconstruct entire organ systems from tissue explants. In Arabidopsis two-step tissue culture system, pluripotency regulators are specifically expressed in the middle-cell layer of the stratified callus tissue. However, regulatory mechanisms underlying the radial patterning of callus remained unclear. Here, we found that very-long-chain fatty acids (VLCFAs) synthesized in the epidermis-like outermost layer are essential for pluripotency acquisition and successful shoot regeneration. Our genetic and transcriptomic analyses revealed that the regulatory roles of VLCFAs on pluripotency acquisition involve inter-cell layer signaling in callus tissue, while they are at least partly independent of ATML1/PDF2 functions and cuticular wax synthesis in the outermost layer. VLCFAs spatially restrict procambium cell identity by non-cell-autonomously suppressing cytokinin signaling, thereby allowing for establishment of the middle-cell layer. We propose that the inhibitory relationships between layer-specific regulators underlie the intricate balance of cellular fate determination in pluripotent callus.

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