on was observed for Glycinin G1, Glycinin G2, and KTI 1 with an average %CV less than 10 and a majority of %CV values being 5–15. Medium technical variation was observed for Beta conglycinin α subunit, Gly m Bd 28k, Glycinin G3, Glycinin G4, and KTI 3 with average %CV of less than 15 and a majority of the %CV values being 5–25.
Genotypic and environmental variation assessed using %CV are shown in Figure 2. From this plot, the amount of variation observed was larger between locations than between cultivars, with KTI 1 being the only exception, showing over onefold greater variation from the cultivar than the location. KTI 3 showed a similar differential in variation but in an opposite direction, with the location providing over threefold greater variation than the cultivar. KTI 1 expression was influenced more by the genotype and KTI 3 was affected more by the environment based on the F statistics and P values (Table 1). KTI 1 expression was the only allergen clearly influenced by the cultivar in this study compared to the other allergens with an F-value of 61.57 (Table 1). Cultivar 92B12 had the highest amount of KTI 1 protein and cultivar 93B15 had the lowest (Figure 3). The expression trend of KTI 3 was similar among the cultivars. The environment had the greatest impact on KTI 3 expression in this study compared to the other allergens, with an F-value of 70.54. Cultivar 92B12 had higher levels of KTI 3 compared to the other three commercial cultivars (Figure 3). However, all the cultivars grown in Ontario had low levels of KTI 3 compared to the other locations. KTI 3 levels were twofold to fourfold– reduced for each cultivar grown in Ontario. These drastic differences in variation between the two variables (cultivar and location) were unique to the KTIs. Four paralogs for the seed storage protein glycinin were individually quantified based upon unique peptides, which revealed a surprising range of expression variation. The most prominently expressed paralog, Glycinin G1, showed 5% variation with respect to cultivar and 10% for location, indicating that neither variable was causing large expression changes, when considering %CV. Glycinin G2 also showed low overall variation, however, the disparity between variation from location and cultivar was large, 15 and 2% respectively. In contrast, one of the two lower abundance forms, Glycinin G3, has the largest percent variation. Location resulted in variation exceeding 60%, which was approximately twice the variation observed among cultivars.
The levels of Glycinin G1 and G2 were affected by the environment and to a lesser extent by the cultivar (Table 1). Kansas and Pennsylvania had significantly higher levels of Glycinin G1 compared to the other locations across four cultivars (Figure 3). Conversely, Ontario had significantly lower levels of Glycinin G2 compared to the other locations across four cultivars. The level of Glycinin G3 was impacted by both the environment and the cultivar, but the environmental factor had a greater impact on the expression of this allergen as indicated by both the F statistics and P value (Table 1). Ontario and Pennsylvania had significantly higher levels of Glycinin G3 compared to the other locations across four cultivars.
The levels of Glycinin G4 were affected by the environment (Table 1); with cultivars grown in Georgia and Ontario having the lowest levels (Figure 3). The difference in cultivar also affected the expression of Glycinin G4 (Table 1). Cultivar 92B12 and 93B15 had the lowest levels of Glycinin G4 across all six locations. The expression level of Gly m Bd 28k and β-conglycinin α subunit were affected mainly by the environment it was grown in (Table 1). Iowa and Kansas had the highest levels of Gly m Bd 28k compared to the other sites across four commercial varieties (Figure 3). In contrast, cultivars grown in Georgia were consistently lower than those grown in Iowa and Kansas (Figure 3). Nevertheless, a lower level of β-conglycinin α subunit was observed in all the cultivars grown in Ontario (Figure 3).
The total allergen conte