title: Programmed cell death promotes male sterility in the functional dioecious Opuntia stenopetala (Cactaceae).
abstract: OBJECTIVE
  The sexual separation in dioecious species has interested biologists for decades; however, the cellular mechanism leading to unisexuality has been poorly understood. In this study, the cellular changes that lead to male sterility in the functionally dioecious cactus, Opuntia stenopetala, are described.
  
  
  METHODS
  The spatial and temporal patterns of programmed cell death (PCD) were determined in the anthers of male and female flowers using scanning electron microscopy analysis and histological observations, focusing attention on the transition from bisexual to unisexual development. In addition, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assays were used as an indicator of DNA fragmentation to corroborate PCD.
  
  
  RESULTS
  PCD was detected in anthers of both female and male flowers, but their patterns differed in time and space. Functionally male individuals developed viable pollen, and normal development involved PCD on each layer of the anther wall, which occurred progressively from the inner (tapetum) to the outer layer (epidermis). Conversely, functional female individuals aborted anthers by premature and displaced PCD. In anthers of female flowers, the first signs of PCD, such as a nucleus with irregular shape, fragmented and condensed chromatin, high vacuolization and condensed cytoplasm, occurred at the microspore mother cell stage. Later these features were observed simultaneously in all anther wall layers, connective tissue and filament. Neither pollen formation nor anther dehiscence was detected in female flowers of O. stenopetala due to total anther disruption.
  
  
  CONCLUSIONS
  Temporal and spatial changes in the patterns of PCD are responsible for male sterility of female flowers in O. stenopetala. Male fertility requires the co-ordination of different events, which, when altered, can lead to male sterility and to functionally unisexual individuals. PCD could be a widespread mechanism in the determination of functionally dioecious species.
authors.affiliation: Laboratorio de Desarrollo en Plantas, Departamento de Biología Comparada, Facultad de Ciencias, UNAM México, DF 04510 México.
authors.forename: Lluvia
authors.initials: L
authors.lastname: Flores-Rentería
authors.forename: Gregorio
authors.initials: G
authors.lastname: Orozco-Arroyo
authors.forename: Felipe
authors.initials: F
authors.lastname: Cruz-García
authors.forename: Florencia
authors.initials: F
authors.lastname: García-Campusano
authors.forename: Isabel
authors.initials: I
authors.lastname: Alfaro
authors.forename: Sonia
authors.initials: S
authors.lastname: Vázquez-Santana
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D002470:Cell Survival; D053938:DNA Fragmentation; D035264:Flowers / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D008540:Meiosis; D008800:Mexico; D008855:Microscopy, Electron, Scanning; D029747:Opuntia / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D051479:Plant Infertility / Q000502:physiology*; D012098:Reproduction
keywords: Cactaceae; Opuntia stenopetala; PCD; TUNEL; dioecy; male sterility; microsporogenesis; programmed cell death; reproductive systems; unisexuality