Flow Cytometric Seed Screening and Seed Genotyping: Methods for Detecting Reproductive Modes in Flowering Plants

Area of Science:

• Plant reproductive biology
• Genetics
• Cytometry

Background:

• Flow cytometry is crucial for estimating ploidy and genome size, and for determining reproductive modes in plants via flow cytometric seed screening (FCSS).
• Existing methods like FCSS have limitations in applicability and resolution for certain reproductive pathways.
• Traditional alternatives involving seed germination and seedling genotyping are time-consuming and may miss endosperm ploidy information.

Purpose of the Study:

• To develop a streamlined method combining flow cytometry and seed genotyping for comprehensive analysis of plant reproductive modes.
• To overcome the limitations of individual methods in accurately characterizing complex reproductive strategies.

Main Methods:

• A novel protocol integrating flow cytometry with genotyping on the same individual seeds.
• Application of both flow cytometric seed screening (FCSS) and seed genotyping to analyze reproductive pathways.

Main Results:

• The combined method provides comprehensive insights into reproductive modes, including various forms of apomixis and automixis.
• It enables accurate assessment of selfing levels irrespective of the reproductive mode.
• The technique successfully identifies pollen donors within the endosperm of apomictic seeds.

Conclusions:

• The integrated flow cytometry and genotyping approach offers a powerful tool for dissecting plant reproductive biology.
• This method enhances the understanding of apomixis, automixis, and selfing dynamics.
• Both protocols, when used independently or combined, are valuable for determining basic reproductive modes.