28 SEED TESTING INTERNATIONAL www.seedtest.org RULES DEVELOPMENT • In India: Tests were all on top of paper using an 8-h light / 16-h dark regime. The tests were set up during the light period. 2. The Petri dishes were then placed into plastic bags in order to prevent water loss during the test and held at 20 ±1 °C. Stacking the Petri dishes on top of each other was avoided. If stacking could not be avoided, it was limited to a stack of two dishes. Care was taken to ensure that the temperature was accurate, and the Petri dishes were not crowded in the incubator. 3. RE for each replicate was recorded after 72 h ±15 min. The criterion of RE was ≥2 mm radicle protrusion. The data was recorded on data sheets provided. 4. After 12 days, for each replicate, a final germination count of normal and abnormal seedlings was recorded (ISTA, 2023), as well as dead seeds. Information received from all laboratories confirmed that the test was completed following the guidelines. The only differences between laboratories were in light regime and, for one laboratory, germination paper, as noted above. The data were analysed using R function ISTAgermMV (2023) developed by the ISTA Statistics Committee. Analysis was completed for: • The seven seed lots tested by the five Indian laboratories (i.e. three seed lots from Nunhems plus four lots from IndoAmerican Hybrid Seeds) • The three seed lots from Nunhems tested by both the European and Indian laboratories (11 laboratories in total) Only the data for the seven seed lots tested by five laboratories in India are included in this abbreviated report. Results Analysis of Data for Seven Seed Lots Tested by Five Laboratories The mean laboratory germination for each lot in all laboratories was above 85% for all lots except lot 4, which had a mean laboratory germination of 51% (data not presented). RE counts after 72 h at 20 °C revealed differences between seed lots in each laboratory (Table 3). In comparing the overall means for seven seed lots in each laboratory, it was revealed that laboratory 3 had the lowest results for RE (Table 3), suggesting that radicles equal to or greater than 2 mm may not have been clearly identified. The seed lot mean from all laboratories identified two seed lots (1 and 6) with the highest RE value and hence the highest vigour, with lot 4 having the lowest vigour (Table 3). The ranking of seed lots was similar within each of the five laboratories (Fig. 1). The low RE that suggested the lowest vigour for lot 4 was, however, largely attributable to its low final normal germination. The greater variability in the RE data for lots 7 and 4 (Fig. 1) is what would be expected for seed lots with low vigour. Tolerances: Considering four replicates of 50 seeds, five results out of 35 were out of tolerance, four of which were from laboratory 3 (lots 3, 4, 5 and 6) and one from laboratory 2 (lot 4). Mixed model for comparing test mean seed lot values: Results from fitting the mixed model are displayed in Table 4, indicating a significant effect of seed lots on RE results. Repeatability and reproducibility: The value for repeatability (⨍𝑟 = 1.06, Table 5) is close to 1.0 and is therefore acceptable. The value of 2.09 for reproducibility (⨍𝑅 , Table 6) is close to that theo- retically derived using the over-dispersion factor between laboratories established by Miles (1963). Lab Lot 1 Lot 2 Lot 3 Lot 4 Lot 5 Lot 6 Lot 7 Lab mean 1 80 56 64 50 66 87 54 65 2 88 93 82 38 78 84 84 78 3 58 68 62 30 30 61 30 48 4 89 86 90 74 82 90 82 85 5 86 83 79 48 58 79 56 70 Lot mean 80 77 75 48 63 80 61 69 Table 3. Mean radicle emergence values for seven seed lots tested in five laboratories; each value is the mean of four replicates of 50 seeds Figure 1. Ranking of the seven seed lots of onion (4 × 50 seeds) Source of variation Sum of squares Mean square Num DF Den DF F value Pr (>F) Lot 2478.16 413.03 6 24 8.58 0.000049 Table 4. Results from fitting the mixed model Table 5: Repeatability estimates for seven lots of Allium cepa using a linear mixed model (repeatability standard deviation) (reproducibility standard deviation) 69 13.43 8.63 6.94 1.06 16.33 𝑝𝑝̅… 𝜎𝜎$"#$ 𝜎𝜎$"%&×"#$ 𝑆𝑆( 𝑓𝑓( 𝑆𝑆)
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