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Behavior of Opuntia ficus-indica (L.) Mill. Heat-Stressed Microspores Under In Vitro Culture Conditions as Evidenced by Microscopic Analysis

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Abstract

When constrained by in vitro culture conditions, microspores from Opuntia ficus-indica (L.) Mill. (Barbary fig) anthers were forced to stall out their gametophytic pathway and switch towards androgenesis. Five microspore stages were characterized based on cellular architecture. A relationship was also established between anthers and bud flower features. Anthers were cultured on three culture media containing 2,4-dichlorophenoxyacetic acid, thidiazuron and gibberellic acid at 22°C for 4 wk, followed by heat shock treatment at 32°C or 42°C for 2 wk, and the stressed material was recultured at 22°C for 24 wk in the dark. When anthers were heat shocked at 32°C and 42°C, the cultivated anthers swelled and burst, followed by the microspores flowing onto the explant surface. Androgenesis occurred directly and indirectly at uni- and binucleate stages. The micromorphology of multicellular, proembryos, globular, torpedo and cotyledonary-like-shaped structures from dehisced anthers was confirmed by environmental scanning electron microscopy. Furthermore, at 42°C, microscopic analysis demonstrated that the non-responsive microspores achieved pollen maturation and more rarely, the emission of a pollen tube. However, 11.4 to 14.4% of the mature pollen grains were converted into pollen embryos. The established system may serve, for the first time, as a protocol to produce microspore embryos in O. ficus-indica. Nevertheless, more efforts are needed to complete the development of diplo-haploid plantlets.

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Correspondence to Badra Bouamama-Gzara.

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Editor: Neftali Ochoa-Alejo

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Bouamama-Gzara, B., Zemni, H., Zoghlami, N. et al. Behavior of Opuntia ficus-indica (L.) Mill. Heat-Stressed Microspores Under In Vitro Culture Conditions as Evidenced by Microscopic Analysis. In Vitro Cell.Dev.Biol.-Plant 56, 122–133 (2020). https://doi.org/10.1007/s11627-019-10032-4

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Keywords

  • Barbary fig
  • Heat shock
  • Microspores
  • Phytohormones
  • Pollen embryos