Imaging Double Fertilization in Maize

Abstract

Sexual reproduction in flowering plants relies on double fertilization, a process marked by two fusion events between the male and female gametes that lead to seed formation. Because this process unfolds within the embryo sac embedded deep inside the ovule, direct observation remains technically demanding, especially in maize, where the large size of female reproductive organs presents additional obstacles. The described method enables high-resolution visualization of cellular events unfolding during maize double fertilization. The approach integrates optimized fixation, clearing and confocal imaging of embryo sacs from ears pollinated with fluorescent pollen marker lines. Precise timing of embryo sac fixation is critical, allowing capture of key events such as pollen peri-germ cell membrane break-down or gamete karyogamy. The protocol provides detailed guidance for ovule dissection, fixation, preparation and renewal of the clearing solution and confocal imaging of embryo sacs. This method offers unprecedented access to the cellular events of double fertilization in maize, establishing a robust framework for studying reproductive processes and supporting future discoveries in plant reproduction.

Plant phenotyping relevance

トウモロコシの二重受精過程を高解像度で可視化する固定・透明化・共焦点 imaging プロトコルが研究の中心であり、植物の生殖状態を取得する方法として該当する。

Code and data availability

This is a methods/protocol paper describing maize ovule dissection, fixation, clearing, and confocal imaging of double fertilization. No public phenotype/trait datasets, author-deposited images, analysis code, or trained models are mentioned. The only URL besides the DOI is Fiji (https://fiji.sc/), a generic thirdparty

No evidence-backed public reproduction asset is currently recorded.