Microspore embryogenesis: cell wall dynamics and reprogramming of cell fate
- Autores: Carolina Camacho Fernández
- Directores de la Tesis: José María Seguí Simarro (dir. tes.), Patricia Corral Martínez (dir. tes.)
- Lectura: En la Universitat Politècnica de València ( España ) en 2021
- Idioma: español
- Tribunal Calificador de la Tesis: M. Pilar Vallés Brau (presid.), Benito José Pineda Chaza (secret.), Nuria Alburquerque Ferrando (voc.)
- Programa de doctorado: Programa de Doctorado en Biotecnología por la Universitat Politècnica de València
- Materias:
- Enlaces
- Tesis en acceso abierto en: RiuNet
- Resumen
Doubled haploids are a great tool for hybrid breeding due to their complete homozygosity achievable in only one in vitro generation. Among the several techniques used to obtain these plants, induction of microspore embryogenesis, via anther or microspore culture, is the most common and efficient approach. Microspore embryogenesis is also an example of totipotency of plant cells due to their ability to reprogram themselves from a gametophytic to a sporophytic pathway, where cells proliferate in an organized way to create a new organism. As in many other in vitro procedures, culture conditions must be optimized in order to increase efficiency. In the present Doctoral Thesis, we used two plant species as experimental systems to study and optimize microspore culture. On one hand, we used eggplant (Solanum melongena) as an example of economically important crop where protocols have still room for optimization. Optimization of cell density and growth regulators demonstrated to be useful to modify the efficiency of eggplant microspore cultures. On the other hand, we used B. napus microspore cultures principally for basic studies since it is widely used as a model to understand cellular processes occurring during this developmental switch. A standardized protocol for Brassica napus microspore culture is detailed, which was used in all the cultures included in this Thesis to explore a series of processes and cellular structures potentially involved in the developmental switch towards embryogenesis. These included endoplasmic reticulum stress, programmed cell death, autophagy, and cell wall structure and composition. We studied in parallel microspore cultures from two B. napus genotypes with different androgenic response cultured in standard conditions and adding Trichostatin A, a epigenetic modulator shown to be beneficial for the androgenic response in some cases. Together, this Thesis represents an advance in the optimization of microspore cultures in these species, and sheds light on the role of some processes within the context of microspore embryogenesis.
