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Targeted CENH3 protein depletion in egg cells enables highly efficient haploid induction
Saravanakumar Somasundaram1, Seda Yaşar2, Jörg Fuchs1
1Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, Corrensstrasse 3, 06466 Seeland, Germany.
Plant Communications
|April 1, 2026
Summary
This study engineered a novel method for plant breeding by depleting CENH3 in egg cells, generating haploids efficiently. This approach accelerates the development of homozygous lines for crop improvement.
Area of Science:
- Plant Science
- Genetics
- Molecular Biology
Background:
- Doubled haploid technology is crucial for rapid plant breeding.
- Current methods for inducing haploids, like manipulating CENH3, lack broad applicability.
- The hypothesis suggests CENH3 asymmetry drives chromosome elimination for haploid induction.
Purpose of the Study:
- To experimentally validate the CENH3 asymmetry hypothesis for haploid induction.
- To develop a broadly applicable and efficient system for engineering haploid inducers.
- To establish a universal platform for haploid induction in diverse crop species.
Main Methods:
- Depleting EYFP- or ALFA-tagged CENH3 in egg cells via proteasomal degradation before fertilization.
- Utilizing a plant-derived E3 ubiquitin ligase for enhanced CENH3 degradation.
- Incorporating a fluorescent seed marker for easy haploid identification.
Main Results:
- Consistently generated paternal wild-type haploids with up to 57% induction frequency.
- Demonstrated effectiveness with in-locus ALFA-tagged CENH3 lines.
- Successfully degraded CENH3 in tomato using the plant-derived E3 ubiquitin ligase.
Conclusions:
- The gametic CENH3-depletion system provides a rational, modular framework for engineering haploid inducers.
- This approach offers a potentially universal platform for haploid induction across various crop species.
- The validated hypothesis opens new avenues for accelerating plant breeding programs.
