The possibility of analyzing chromatin topology in developing plant embryos is hampered by inaccessibility of the embryo sac, deeply embedded in the maternal seed tissue, following double fertilization. Here we describe a protocol to isolate, purify, and prepare developing Boechera stricta embryos for chromosome conformation capture-based methods as in situ Hi-C experiments. Early globular embryos can be isolated by air-pressure microaspiration, and subsequently washed to eliminate residual cells from the endosperm and maternal seed coat, allowing for pure sampling of selected stages of embryogenesis. This protocol allows for the possibility of comparing genome topology during plant embryonic differentiation since early until late embryo development stages.
Keywords: Apomixis; Boechera stricta; Chromosome conformation capture; Embryo isolation; Flowering plants embryo development; Genome topology; Hi-C.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.