The involvement of programmed cell death in inflated leaf petiole morphogenesis in Trapa pseudoincisa.

Trapa plants (Trapaceae) have an inflated leaf petiole called a spongy airbag. The aims of this study were to assess the involvement of programmed cell death (PCD) in the process of inflated leaf petiole morphogenesis. In this paper, light and transmission electron microscopy (TEM) were used to investigate cytological events and the development of inflated leaf petiole. During this process, the inflated leaf petiole of Trapa pseudoincisa L. undergoes a developmental process, changing from solid to hollow phase. Debris from the degraded cells was seldom observed in the transverse sections of leaf petioles, but some degraded cells with an abnormal morphology were observed in longitudinal sections. Cytoplasmic changes, such as disrupted vacuoles, degraded plastids, and the emergence of secondary vacuoles were observed during leaf petiole morphogenesis. In addition, gel electrophoresis and TUNEL assays were used to evaluate DNA cleavage during petiole morphogenesis. DNA internucleosomal cleavage and TUNEL-positive nuclei indicate that the typical PCD features of DNA cleavage occurred early in the process. These results revealed that PCD plays a critical role in inflated leaf petiole morphogenesis. Additionally, a trans-disciplinary systems approach is required that recognises the necessity for integration of cytological and molecular characteristics for identification of aerenchyma type.