Programmed cell death (PCD) is an essential process for development, and shows conserved cytological features in both plants and animals. Caspases are well‐known critical components of the PCD machinery in animals. However, currently few typical counterparts have been identified in plants and only several caspase‐like proteases are known to be involved in plant PCD, indicating the existence of great challenge for confirming new caspase‐like proteases and elucidating the mechanisms regulating plant PCD. Here, we report a novel cysteine protease, NtTPE8, which was extracted from tobacco seeds and confirmed as a new caspase‐like protease. Recombinant NtTPE8 exhibited legumain and caspase‐like proteolytic activities, both of which could be inhibited by the pan‐caspase inhibitor (Z‐VAD‐FMK). Notably, NtTPE8 possessed several caspase activities and the capacity to cleave the cathepsin H substrate FVR, indicating a unique character of NtTPE8. NtTPE8 was exclusively expressed in the integumentary tapetum and thus, is the first specific molecular marker reported to date for this cell type. Down‐regulation of NtTPE8 caused seed abortion, via disturbing early embryogenesis, indicating its critical role in embryogenesis and seed development. In conclusion, we identified a novel caspase‐like cysteine protease, NtTPE8, exclusively expressed in the integumentary tapetum that is involved in seed development.

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