Ovaries and anthers collected at pre-anthesis stages (white or pale-yellow anthers) from independent T2 plants of apomictic events T314 15.1 and T314 37.7 (and of replicated samples collected on T3 plants of event T314 15.1) were fixed in Carnoy’s or FAA fixatives and rinsed in 70% ethanol. Dissected ovaries were mounted in Hoyer’s clearing medium and gently squashed by pressure on the coverslip to expose ovules. Callose detection using aniline blue staining was performed as described60 (link). Observation and imaging were done on a Zeiss Axioimager Z2 microscope equipped with DIC, CFP emission filter; ×40 or ×63 (NA1,4) oil immersion lenses, and a sCMOS camera (Hamamatsu ORCA Flash V2). Images were processed using ImageJ software.
Vernet A., Meynard D., Lian Q., Mieulet D., Gibert O., Bissah M., Rivallan R., Autran D., Leblanc O., Meunier A.C., Frouin J., Taillebois J., Shankle K., Khanday I., Mercier R., Sundaresan V, & Guiderdoni E. (2022). High-frequency synthetic apomixis in hybrid rice. Nature Communications, 13, 7963.
Other organizations :
Centre de Coopération Internationale en Recherche Agronomique pour le Développement, Max Planck Institute for Plant Breeding Research, Council for Scientific and Industrial Research, Université de Montpellier, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement, Institut Agro Montpellier, Institut de Recherche pour le Développement, Diversité, adaptation et développement des plantes, University of California, Davis
Apomictic events T314 15.1 and T314 37.7 (and replicated samples collected on T3 plants of event T314 15.1)
dependent variables
Callose detection using aniline blue staining
control variables
Ovaries and anthers collected at pre-anthesis stages (white or pale-yellow anthers)
Ovaries were fixed in Carnoy's or FAA fixatives and rinsed in 70% ethanol
Dissected ovaries were mounted in Hoyer's clearing medium and gently squashed by pressure on the coverslip to expose ovules
Observation and imaging were done on a Zeiss Axioimager Z2 microscope equipped with DIC, CFP emission filter; ×40 or ×63 (NA1,4) oil immersion lenses, and a sCMOS camera (Hamamatsu ORCA Flash V2)
controls
No positive or negative controls were explicitly mentioned in the provided information.
Annotations
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