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34. The session now moves into the Q&A, facilitated by Andreas Wais, @ISTAseedtesting Secretary General. He thanks the speakers for their detailed presentations and invites questions from participants.

33. She closes with practical recommendations: ✅ Use a sharp scalpel (change after ~100 seeds) ✅ Smaller seeds hydrate better between filter papers ✅ When testing ungerminated seeds at the end of a germination test, no pretreatment is required.

32. In Festuca rubra, multiple-seed units are treated as one sample. Stefanie shows viable and non-viable examples, noting how incomplete cuts can lead to misinterpretation.

31. For Festuca rubra, she explains two approaches: – Longitudinal cut (2 h at 30 °C) – Transversal cut (18 h at 30 °C) Both use 1 % TZ solution.

30.🔍 Evaluation criteria A seed is considered viable if no more than: – ⅓ of the radicle (from the tip) and – ⅓ of the scutellum show unstained or flaccid tissue. If uncertain, the embryo can be excised to observe the back of the scutellum.

29. 🧪 Staining process: 2 hours at 30 °C in 1.0 % TZ solution. After staining, the two halves are spread to expose the cut surface for evaluation.

28. ✂️ Preparation before staining. Stefanie demonstrates how to: – Cut longitudinally through the embryo and ¾ of the endosperm – Or, if glumes can be removed, cut transversely near the embryo.

27. 💧Pretreatment: Seeds are soaked for 3 hours at 20 °C or 16 hours between moist filter paper. This ensures full hydration before staining.

26. She presents the essential instruments for the test: Breakers, bowls, razor blades, scalpels, filter paper, forceps, dissecting needles, and a binocular microscope. “Clean, sharp tools are critical to avoid cutting damage,” Stefanie reminds participants.

25. Next up: Stefanie Krämer. She begins by introducing the species under study: 🌱 Lolium spp. 🌱 Festuca rubra 🌱 Poa spp. She references ISTA Rules Table 6A, explaining the seed’s internal structure. Glume, endosperm, scutellum, and embryo.

24. Eoin concludes this section by comparing heat-damaged barley seeds in a germination test. They often sprout weakly, producing short roots or soft, dead seedlings.

23. Stage IV: Advanced heat damage. Full embryo necrosis, complete loss of viability.

22. Stage III: Severe internal damage. Most tissues lose staining capacity.

21. Stage II: Pinkish, milky colouration appears. Tissues start to deteriorate.

20. Eoin explains that heat damage progresses through four stages (I–IV).

19. Heat damage now under discussion. This occurs during drying or storage, especially when “hot spots” form in damp lots, killing cells from the centre outward.