Supplementary Tables and Figures

 

Table S1. Significance values of principal components (PCs) for ontogeny, subspecies and colony. Significance (p-value) of experimental parameters is indicated for the first 9 PCs. Ontogeny was primarily associated with PC1 and 2, while subspecies was primarily associated with PC3. Although PC1 and 2 were strongly associated with pre-foraging maturation and the hive bee to forager transition, they were not associated with age differences within hive bees after 8 days of age (p > 0.1). Colony was significant for several PCs within the mature hive bee group and probably corresponded to environmental differences. Indicated p-values were corrected using a conservative family-wise adjustment for the 9 PCs tested (Holmes). Multivariate tests of global gene expression (‡) were conducted using the first 9 PCs. The first 9 PCs were retained for analyses based on 3 criteria: explanation of arbitrary threshold of variance (80%); retention of PCs with eigenvalues greater than the average eigenvalue; and inspection of variance plot (Fig. 1a) for a natural break between “large” and “small” eigenvalues.

 

 

				All 72 beesa			Mature hive bees (Fig. 1b)b
PC	Var.	Cum. Var.		Ontogeny	Sub-species		Ontogeny	Sub-species	Colony
1	34.4%	34.4%		<2e-15	0.0007		0.90	0.018	1.0
2	18.9%	53.3%		<2e-15	0.007		0.12	0.027	0.0008
3	11.9%	65.2%		8e-11	<2e-15		0.002	<2e-15	0.0009
4	5.0%	70.2%		0.045	1.0		1.0	1.0	0.063
5	3.0%	73.2%		1.0	1.0		1.0	1.0	0.005
6	2.5%	75.7%		1.0	1.0		1.0	1.0	2e-5
7	2.1%	77.8%		0.0004	1.0		0.0005	1.0	0.006
8	1.7%	79.4%		1.0	1.0		0.30	1.0	1.0
9	1.3%	80.8%		0.0005	1.0		0.054	1.0	1.0
1-9c	80.8%	80.8%		<2e-15	<2e-15		0.0004	2e-12	4e-7

^aTwo-way ANOVA with factors ontogeny and subspecies, using 72 bees collected from field colonies; ontogeny df = 5; subspecies df = 1. The interaction term, ontogeny × subspecies, was marginally significant only for PC3 (p = 0.051).

^bThree-way ANOVA with factors ontogeny, subspecies, and trial, using only the 29 “mature hive bees” from Fig. 1b; ontogeny: df = 2; subspecies df = 1; host colony df = 1. Several significant interactions were detected in PCs 5 – 9 (p < 0.05).

^cMultivariate ANOVA (MANOVA) using PC1 – 9; Pillai-Bartlett statistic. For 2-way MANOVA: ontogeny df0 = 45, df1 = 280; subspecies df0 = 9, df1 = 52; the interaction term, ontogeny × subspecies, was marginally significant (p = 0.055). For 3-way MANOVA: ontogeny df0 = 18, df1 = 22; subspecies df0 = 9, df1 = 10; host colony df0 = 9, df1 = 10; significant interactions were subspecies × host colony (p = 0.0007) and ontogeny × host colony (p = 0.031).