Optimizing Visualization of Pollen Tubes in Wheat Pistils.

Abstract

Successful pollination and fertilization are crucial for grain setting in cereals. Wheat is an allohexaploid autogamous species. Due to its evolutionary history, the genetic diversity of current bread wheat ( Triticum aestivum ) cultivars is limited. Introducing favorable alleles from related wild and cultivated wheat species is a promising breeding strategy for resolving this issue. However, wide hybridization between bread wheat and its relatives is hampered by the presence of suppressor genes and difficulties in crossing. Optimized methods for observing pollen tubes are essential for understanding the mechanism of crossability between wheat and its relatives. Here, we improved the crossing procedure between bread wheat and rye ( Secale cereale ) and established an optimized protocol for visualizing pollen tube behavior. Crossing via detached spike culture significantly enhanced crossing efficiency and phenotypic stability. A combination of canonical aniline blue staining and optimized clearing and sectioning allowed us to visualize pollen tube behavior. The proportion of rye pollen tubes reaching the micropyle was lower than that for pollen tubes germinated on the stigmatic hair, explaining why the hybrid seed-setting rate was approximately 75% instead of 100%. This method sheds light on wide hybridization through deeper visualization of the insides of pistils.

Plant phenotyping relevance

コムギ雌ずい内の花粉管挙動を可視化する染色・透明化・切片化手法の最適化が研究の中心であり、植物の生殖状態を測定する実質的な表現型取得法に該当する。

Code and data availability

植物フェノタイピング解析を再現する公開資産であることを、入力本文と直接リンクから確認できなかったため保留しました。

No evidence-backed public reproduction asset is currently recorded.