26 SEED TESTING INTERNATIONAL www.seedtest.org RULES DEVELOPMENT • showed 37% fresh seed on average in the two laboratories for the standard germination method, 8% for the preheating method, 16% for the prechilling method and 5.5% for the ethephon method. The same observations were made for lot H10, although the seeds were less dormant. On average for the two lots with dormancy, the ethephon method gives similar results to the preheating method and significantly better results than the prechilling method. Repeatability and reproducibility estimates are presented in Table 2. The dispersion factor is below 1 only for the ethephon method, indicating good repeatability of the results obtained with this method when dormancy is observed. Repeatability is therefore better than for the other two methods of dormancy breaking. Conclusion This study shows the value of adding the dormancy-breaking method using ethephon seed treatment to the ISTA Rules. When dormancy was observed, two laboratories demonstrated the effectiveness of the ethephon method. This method is just as effective as the preheating method and better than the prechilling method. Across all lots and laboratories, the ethephon method was found to be as effective as the two official methods for dormancy breaking. Given the preliminary studies showing the effectiveness of the ethephon method (Ducournau and Dupont, 2023, 2024), especially on highly dormant sunflower seed samples, we propose to add this new method to the ISTA Rules. Acknowledgements We gratefully acknowledge seed companies Limagrain and Corteva for supplying the seed lots. Many thanks to all the laboratories that participated in this validation study and thanks to the reviewers from ISTA Technical Committees. Thanks to Jean-Louis Laffont (ISTA Statistics Committee) who helped run the data and for the support he gave in statistical interpretation. References 1. Corbineau, F., Xia, Q., Bailly, C. and El- Maarouf-Bouteau, H. (2014). Ethylene, a key factor in the regulation of seed dormancy. Frontiers in Plant Science, 5. 2. Ducournau, S. and Dupont, A. (2023). Dormancy breaking methods for sunflower seeds. Reports of the ISTA Annual Meeting, 29 May – 1 June 2023, Verona, Italy. International Seed Testing Association, Wallisellen, Switzerland. 3. Ducournau, S. and Dupont, A. (2024). Dormancy breaking methods for sunflower seeds. Reports of the ISTA Annual Meeting, 1–4 July 2024, Cambridge, UK. International Seed Testing Association, Wallisellen, Switzerland. 4. Maiti, R.K., Vidyasagar, P., Shahapur, S.C, Ghosh, S.K. and Seiler, G.J. (2006). Development and standardization of a simple technique for breaking seed dormancy in sunflower (Helianthus annuus L.). Helia, 29(45), 117–126. Figure 3. Boxplots for the four methods per dormancy seed lot (lot H7 and lot H10) and grouped by laboratory on fresh seed Table 2. Repeatability and reproducibility of the different methods, without laboratory E Method Mean s_Repeatability Disp S_Reproducibility s_Lab s_Lot × Lab ETHREL 85 2.84 0.79 10.20 8.50 5.26 PRECHILLING 81 4.38 1.12 6.19 4.88 2.21 PREHEATING 85 4.09 1.14 2.89 0.00 0.00 SD 73 5.48 1.23 10.63 8.93 4.27
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