From the Cover: A diapause pathway underlies the gyne phenotype in Polistes wasps, revealing an evolutionary route to caste-containing insect societies -- Hunt et al. 104 (35): 14020 Data Supplement - HTML Page - index.htslp -- Proceedings of the National Academy of Sciences

Hunt et al. 10.1073/pnas.0705660104.

Supporting Information

Files in this Data Supplement:

SI Table 1
SI Table 2
SI Table 3
SI Table 4
SI Table 5
SI Figure 6
SI Table 6

SI Figure 6

Fig. 6. Coomassie stained SDS/PAGE of Polistes metricus hemolymph proteins. Hemolymph from adult gynes was collected in extraction buffer. After methanol/chloroform precipitation and solvation in loading buffer, proteins were subjected to SDS./PAGE. Molecular mass standards are in the left lane, and the P. metricus proteins are in the right lane. The numbered coomassie-stained bands were cut out of the gel, and the proteins in them were identified by LC-MS/MS after in-gel digestion using the protease trypsin. The software programs OMSSA and NovoHMM and manual inspection were used to determine valid identifications as listed in Table 6. Only proteins identified by at least two peptides are listed.

Table 1. Dates of cocoon spinning, adult emergence, and pupation duration in days for foundress-reared (primarily worker destined) and worker-reared (primarily gyne destined) female offspring in nests in the field

Nest, no.	Cocoon	Emergence	Duration
Foundress-Reared
1	31 May	18 Jun	18
1	1 Jun	20 Jun	19
16	1 Jun	19 Jun	18
46	1 Jun	20 Jun	19
46	1 Jun	20 Jun	19
16	2 Jun	20 Jun	18
31	2 Jun	20 Jun	18
67	2 Jun	22 Jun	20
1	3 Jun	22 Jun	19
46	4 Jun	23 Jun	19
1	5 Jun	23 Jun	18
1	5 Jun	24 Jun	19
1	5 Jun	24 Jun	19
1	5 Jun	24 Jun	19
16	5 Jun	23 Jun	18
16	5 Jun	24 Jun	19
31	5 Jun	25 Jun	20
46	5 Jun	24 Jun	19
67	5 Jun	24 Jun	19
1	6 Jun	26 Jun	20
1	6 Jun	26 Jun	20
16	6 Jun	25 Jun	19
31	6 Jun	26 Jun	20
67	6 Jun	26 Jun	20
1	8 Jun	27 Jun	19
31	8 Jun	27 Jun	19
Worker-Reared
1	3 Jul	22 Jul	19
1	3 Jul	22 Jul	19
46	3 Jul	23 Jul	20
62	3 Jul	23 Jul	20
142	3 Jul	22 Jul	19
7	4 Jul	23 Jul	19
8	4 Jul	23 Jul	19
8	4 Jul	23 Jul	19
8	4 Jul	24 Jul	20
46	4 Jul	23 Jul	19
46	4 Jul	23 Jul	19
1	5 Jul	23 Jul	18
62	5 Jul	24 Jul	19
1	6 Jul	24 Jul	18
25	6 Jul	25 Jul	19
49	6 Jul	25 Jul	19
46	7 Jul	25 Jul	18
67	8 Jul	26 Jul	18

Nest numbers <100 were at Tyson; numbers >100 were at Shaw. Jul, July; Jun, June.

Table 2. Mean daily temperature maxima and minima and the medians of the two (^oC + SE) during development from cocoon spinning to adult emergence (in the field) or adult molt (in the lab) for foundress-reared and worker-reared offspring

Offspring	High	Low	Mean
F-R field	30.1 + 0.6	15.8 + 0.5	23.0 + 0.5
W-R field	32.9 + 0.8	19.3 + 0.6	26.1 + 0.6
F-R lab	27.8 + 0.2	21.0 + 0.4	24.4 + 0.2
W-R lab	26.9 + 0.1	20.9 + 0.1	24.0 + 0.1

F-R, foundress-reared; W-R worker-reared.

Table 3. Development times in days by developmental stage for foundress-reared (primarily worker destined) and worker-reared (primarily gyne destined) female offspring collected in the field on the day of cocooning and reared in the lab

Specimen / cocoon date	Stage durations, days
Specimen / cocoon date	Larva	Prepupa	Early Pupa	Late Pupa	Total
Lab development, foundress-reared female offspring
B / 3 Jun	1	3	7	5	16
C / 3 Jun	1	3	7	4	15
D / 3 Jun	1	3	8	5	17
E / 4 Jun	1	3	8	4	16
G / 4 Jun	1	3	7	4	15
H / 4 Jun	2	3	7	5	17
I / 5 Jun	1	3	7	4	15
J / 5 Jun	1	2	7	4	14
K / 5 Jun	2	3	8	4	17
M / 6 Jun	1	3	7	4	15
N / 6 Jun	1	3	7	5	16
P / 6 Jun	1	3	7	4	15
Q / 7 Jun	1	3	8	4	16
R / 7 Jun	1	3	9	3	16
S / 7 Jun	1	3	8	5	17
T / 7 Jun	1	3	8	4	16
U / 8 Jun	1	2	9	4	16
V / 9 Jun	1	3	9	4	17
Mean	1.1	2.9	7.7	4.2	15.9
Lab development, worker-reared
GA / 6 Jul	1	4	11	5	21
GC / 6 Jul	2	4	10	5	21
GD / 6 Jul	2	4	9	6	21
GG / 7 Jul	1	4	10	6	21
GH / 7 Jul	2	4	9	6	21
GK / 8 Jul	2	4	9	6	21
GU / 13 Jul	1	4	10	6	21
GW / 13 Jul	2	4	9	5	20
HH / 18 Jul	2	4	10	5	21
HL / 19 Jul	1	4	10	5	20
HX / 20 Jul	1	4	10	6	21
HY / 20 Jul	1	4	10	5	20
Mean	1.5	4	9.8	5.5	20.8

Table 5. Specimen collection and storage protein data

STAGE	F/W	SPM	NEST	COCOON	COLLECT	AGE	HEX1	HEX2	HiGLX	VHDL
L	F	5	69	05 VI	06 VI	2	15.2	62.1	7.1	46.4
L	F	7	145	04 VI	06 VI	3	6.4	45.7	7.5	40.3
L	F	8	146	05 VI	06 VI	2	6.8	37.7	11.7	38.0
L	F	9	146	02 VI	06 VI	5	5.3	21.0	1.9	20.6
L	F	14	150	04 VI	07 VI	4	4.6	25.0	24.5	23.0
L	F	16	156	06 VI	07 VI	2	18.1	47.7	10.0	34.5
L	F	18	3	08 VI	08 VI	1	6.5	31.2	13.4	25.0
L	F	19	5	08 VI	08 VI	1	11.7	47.3	9.6	17.5
L	F	23	133	08 VI	08 VI	1	7.2	52.6	13.3	29.7
L	F	25	158	07 VI	08 VI	2	19.7	55.9	4.0	40.2
L	F	26	158	09 VI	09 VI	1	22.6	27.2	12.6	19.0
L	F	33	138	07 VI	09 VI	3	33.5	22.4	23.5	32.5
L	F	38	110	08 VI	10 VI	3	16.4	22.8	9.8	23.6
L	F	39	123	07 VI	10 VI	4	10.7	22.3	15.7	15.4
L	F	42	2	11 VI	13 VI	3	31.3	21.0	6.8	40.9
P1	F	6	124	02 VI	06 VI	5	2.7	16.4	2.4	9.4
P1	F	10	2	04 VI	07 VI	4	9.2	28.8	5.4	18.7
P1	F	11	32	03 VI	07 VI	5	6.7	20.9	5.7	9.8
P1	F	12	32	31 V	07 VI	8	10.2	30.4	14.9	9.8
P1	F	13	32	02 VI	07 VI	6	9.9	35.3	10.3	19.2
P1	F	15	150	02 VI	07 VI	6	6.7	13.1	2.5	4.5
P1	F	17	156	01 VI	07 VI	7	13.4	60.3	26.8	35.9
P1	F	20	23	01 VI	08 VI	8	5.5	20.1	3.4	7.2
P1	F	21	35	31 V	08 VI	9	6.1	22.6	13.6	8.6
P1	F	22	132	02 VI	08 VI	7	4.7	24.2	22.7	14.4
P1	F	24	157	02 VI	08 VI	7	7.1	20.5	22.0	17.3
P1	F	27	23	04 VI	09 VI	6	7.7	42.2	13.5	27.5
P1	F	28	35	31 V	09 VI	10	8.8	16.2	2.9	8.6
P1	F	30	106	03 VI	09 VI	7	8.4	36.5	19.7	26.0
P1	F	31	112	04 VI	09 VI	6	12.6	21.3	14.2	15.4
P1	F	32	113	01 VI	09 VI	9	10.6	13.7	11.6	9.8
P1	F	34	2	07 VI	10 VI	4	12.4	18.0	12.8	14.9
P1	F	35	4	02 VI	10 VI	9	9.8	43.6	14.6	15.9
P1	F	36	4	01 VI	10 VI	10	23.7	25.3	23.2	13.9
P1	F	37	32	31 V	10 VI	11	31.2	22.8	13.9	6.9
P1	F	40	133	01 VI	10 VI	10	8.9	35.4	22.8	20.6
P1	F	41	157	06 VI	10 VI	5	8.3	34.7	23.3	23.9
P1	F	46	29	05 VI	15 VI	11	11.2	15.2	14.1	11.6
P1	F	47	29	05 VI	15 VI	11	9.0	21.5	16.2	9.3
P1	F	48	65	08 VI	15 VI	8	7.3	11.2	5.3	8.3
P1	F	49	65	07 VI	15 VI	9	3.2	6.2	6.2	5.3
P1	F	50	156	08 VI	15 VI	8	5.5	28.5	22.2	15.9
P1	F	51	157	06 VI	15 VI	10	20.6	14.7	10.7	8.3
P1	F	55	144	06 VI	16 VI	11	20.1	21.4	22.9	7.8
P2	F	44	29	04 VI	15 VI	12	4.0	18.0	6.1	6.5
P2	F	45	29	01 VI	15 VI	15	10.4	13.3	5.1	3.5
P2	F	52	132	01 VI	16 VI	16	5.7	5.6	4.6	5.3
P2	F	53	133	04 VI	16 VI	13	7.9	14.1	10.0	1.5
P2	F	54	144	02 VI	16 VI	15	24.0	26.8	7.5	6.1
P2	F	56	150	05 VI	16 VI	12	15.9	16.6	15.5	10.2
P2	F	57	155	02 VI	16 VI	15	13.9	17.8	4.4	2.8
P2	F	58	155	05 VI	16 VI	12	8.3	25.5	13.9	12.3
P2	F	59	155	05 VI	16 VI	12	4.7	9.8	5.7	6.5
P2	F	61	15	01 VI	19 VI	15	2.2	9.0	1.3	2.0
P2	F	62	23	02 VI	19 VI	18	20.9	22.7	2.3	3.6
P2	F	63	15	02 VI	19 VI	18	5.9	11.4	1.4	2.7
P2	F	64	2	08 VI	20 VI	13	11.2	12.0	8.2	4.8
P2	F	65	4	05 VI	20 VI	16	12.3	18.7	7.0	5.6
P2	F	66	2	04 VI	20 VI	17	17.6	11.9	7.0	5.8
P2	F	67	2	04 VI	20 VI	13	5.9	11.4	4.0	0.8
P2	F	68	65	07 VI	20 VI	14	13.7	20.5	10.8	6.6
P2	F	69	125	08 VI	20 VI	13	7.9	15.6	8.7	1.6
P2	F	70	125	08 VI	20 VI	18	10.2	16.4	2.2	2.3
P2	F	71	3	08 VI	21 VI	14	6.0	19.8	16.3	8.1
P2	F	72	5	06 VI	21 VI	16	12.7	17.4	16.6	6.5
P2	F	73	5	06 VI	21 VI	16	11.8	24.6	11.7	4.4
P2	F	74	5	08 VI	21 VI	14	28.8	5.3	23.5	4.9
P2	F	75	5	05 VI	21 VI	17	8.6	7.1	3.7	2.6
P2	F	76	5	05 VI	21 VI	18	1.7	2.7	0.0	0.0
P2	F	77	64	08 VI	21 VI	14	9.9	28.4	19.9	15.6
P2	F	81	22	08 VI	22 VI	15	3.0	6.1	5.2	2.1
P2	F	82	22	05 VI	21 VI	17	6.3	9.8	2.3	1.4
P2	F	84	22	05 VI	21 VI	17	5.1	4.6	2.2	3.0
P2	F	87	69	05 VI	23 VI	19	7.3	9.0	0.9	0.6
A	F	60	123	01 VI	19 VI	19	0.7	1.0	0.6	0.6
A	F	78	22	04 VI	22 VI	19	0.4	0.8	0.2	0.3
A	F	79	22	04 VI	22 VI	19	0.9	1.7	0.9	1.2
A	F	80	22	04 VI	22 VI	19	0.5	1.4	0.0	0.0
A	F	85	64	04 VI	23 VI	20	0.3	3.0	0.0	0.1
A	F	86	65	04 VI	23 VI	20	0.4	1.3	0.3	0.4

L	W	88	115	04 VII	08 VII	5	11.4	34.3	11.8	35.0
L	W	89	119	05 VII	08 VII	4	5.4	46.1	6.1	30.4
L	W	93	141	07 VII	10 VII	4	5.2	38.1	5.9	22.3
L	W	94	160	06 VII	10 VII	5	7.9	40.8	9.1	17.7
L	W	95	136	07 VII	11 VII	5	31.5	17.2	6.8	13.6
L	W	127	62	20 VII	21 VII	2	16.2	52.7	5.3	52.6
L	W	133	139	20 VII	22 VII	3	10.2	40.4	4.1	12.8
L	W	154	20	21 VII	24 VII	4	9.1	45.9	29.5	29.9
L	W	155	20	23 VII	24 VII	2	14.3	72.7	20.5	47.0
L	W	156	115	24 VII	24 VII	1	51.9	67.5	11.0	28.0
L	W	164	25	23 VII	25 VII	3	11.2	27.7	21.8	13.3
L	W	166	53	24 VII	25 VII	2	30.7	12.3	23.3	12.9
L	W	174	129	23 VII	26 VII	4	13.4	29.1	14.8	18.7
L	W	175	136	24 VII	26 VII	3	10.6	22.2	17.8	24.3
L	W	194	115	25 VII	28 VII	4	17.5	25.6	19.5	13.3
L	W	196	19	25 VII	29 VII	5	7.2	31.6	6.0	13.1
L	W	198	48	26 VII	29 VII	4	25.6	18.4	5.8	14.2
P1	W	96	141	04 VII	11 VII	8	1.1	42.8	8.1	22.2
P1	W	97	119	07 VII	12 VII	6	22.4	2.9	5.7	7.5
P1	W	98	119	04 VII	12 VII	9	46.0	10.6	14.2	10.6
P1	W	99	119	05 VII	12 VII	8	4.6	24.6	6.2	9.8
P1	W	100	20	04 VII	13 VII	10	9.5	30.6	9.0	24.8
P1	W	101	20	05 VII	13 VII	9	20.9	19.5	12.1	20.1
P1	W	104	48	04 VII	14 VII	11	15.4	11.1	6.7	10.0
P1	W	105	49	07 VII	14 VII	8	17.5	17.8	16.7	16.6
P1	W	106	51	05 VII	14 VII	10	5.3	8.6	6.1	5.1
P1	W	109	142	10 VII	16 VII	7	11.9	23.6	10.1	13.7
P1	W	110	149	07 VII	16 VII	10	6.3	17.8	7.3	10.9
P1	W	137	142	13 VII	23 VII	11	26.7	7.2	15.6	10.0
P1	W	138	129	15 VII	23 VII	9	31.4	12.2	9.8	6.1
P1	W	139	142	16 VII	23 VII	8	8.0	13.3	18.2	7.7
P1	W	140	142	18 VII	23 VII	6	14.2	20.5	10.8	12.2
P1	W	153	119	17 VII	24 VII	8	10.6	20.1	15.4	13.1
P1	W	160	53	17 VII	25 VII	9	30.6	38.9	10.8	9.8
P1	W	161	25	20 VII	25 VII	6	5.8	13.7	5.7	6.0
P1	W	171	129	17 VII	26 VII	10	26.0	32.9	7.2	17.1
P1	W	172	19	20 VII	26 VII	7	14.2	33.7	24.6	17.8
P1	W	173	129	22 VII	26 VII	5	6.3	9.0	1.8	5.3
P1	W	179	119	17 VII	27 VII	11	6.3	10.9	3.6	3.8
P1	W	180	119	21 VII	27 VII	7	32.7	39.6	13.9	17.1
P1	W	181	142	22 VII	27 VII	6	13.5	40.4	25.5	24.6
P1	W	183	20	23 VII	27 VII	5	10.2	36.2	15.0	14.3
P1	W	184	48	23 VII	27 VII	5	6.7	13.3	2.8	4.8
P1	W	190	160	18 VII	28 VII	11	14.9	18.9	2.8	2.0
P1	W	191	115	23 VII	28 VII	6	18.0	19.5	12.4	12.1
P1	W	192	160	24 VII	28 VII	5	18.2	17.6	11.6	10.2
P2	W	107	142	05 VII	16 VII	12	14.9	23.7	16.0	13.5
P2	W	108	142	03 VII	16 VII	14	8.6	20.3	7.9	6.8
P2	W	111	20	03 VII	17 VII	15	10.0	14.5	4.0	3.1
P2	W	112	56	04 VII	18 VII	15	15.9	18.9	9.3	7.4
P2	W	113	20	03 VII	19 VII	17	11.2	21.6	7.1	13.6
P2	W	114	7	04 VII	19 VII	16	20.6	18.8	9.8	6.9
P2	W	115	20	05 VII	19 VII	15	24.2	17.1	4.4	4.0
P2	W	116	62	03 VII	20 VII	18	13.3	25.3	0.8	0.7
P2	W	117	19	04 VII	20 VII	17	29.4	10.6	4.4	2.4
P2	W	118	19	05 VII	20 VII	16	26.3	12.3	2.9	2.4
P2	W	119	19	07 VII	20 VII	14	5.9	11.3	4.3	2.2
P2	W	121	48	04 VII	21 VII	18	36.4	9.9	0.3	1.2
P2	W	122	62	05 VII	21 VII	17	17.4	23.8	1.1	1.5
P2	W	123	62	06 VII	21 VII	16	19.4	20.7	5.1	3.6
P2	W	124	62	07 VII	21 VII	15	18.4	16.1	8.6	4.8
P2	W	125	62	09 VII	21 VII	13	8.8	18.7	8.6	5.6
P2	W	126	48	10 VII	21 VII	12	13.2	17.0	12.8	5.7
P2	W	129	142	05 VII	22 VII	18	5.1	6.5	0.3	0.7
P2	W	130	141	06 VII	22 VII	17	35.1	33.7	0.5	1.4
P2	W	131	119	07 VII	22 VII	16	38.9	16.9	2.4	0.9
P2	W	132	8	10 VII	22 VII	13	23.5	25.9	4.5	3.0
P2	W	132	8	10 VII	22 VII	13	32.3	34.4	9.4	9.0
P2	W	152	8	11 VII	24 VII	14	16.0	20.7	6.5	3.4
P2	W	169	139	13 VII	26 VII	14	40.2	20.7	0.3	1.8
P2	W	170	139	14 VII	26 VII	13	6.2	11.6	4.2	15.8
P2	W	189	141	16 VII	28 VII	13	12.1	23.9	7.4	7.4
P2	W	206	48	19 VII	30 VII	12	12.9	12.2	0.5	0.0
P2	W	207	139	19 VII	30 VII	12	13.0	26.7	5.7	6.9
A	W	120	48	03 VII	21 VII	19	15.0	15.4	0.6	0.9
A	W	128	7	04 VII	22 VII	19	1.0	7.3	0.2	0.7
A	W	135	62	03 VII	23 VII	21	3.1	2.2	1.3	1.5
A	W	136	115	04 VII	23 VII	20	6.5	3.0	1.6	1.7
A	W	151	119	07 VII	24 VII	18	6.4	10.2	0.8	0.5
A	W	158	49	06 VII	25 VII	20	0.5	3.0	0.2	0.4
A	W	159	46	07 VII	25 VII	19	1.5	9.5	0.6	0.8
A	W	195	25	11 VII	29 VII	19	8.1	2.7	0.4	0.5

Stages are L, fifth instar larva (both pre-defecation and post-defecation, the "prepupa"); P1, pupa up to day 11 of pupation; P2, pupa from day 12 of pupation; A, adult, including preemergence adults in nest cells and adults collected on the morning after emergence.

F/W: F, foundress reared; W, worker-reared. SPM, individual; NEST, colony. Nests numbered <100 were at Tyson, and nests numbered >100 were at Shaw. COCOON and COLLECT are dates of spinning and collection, all 2005. AGE is in days from COCOON to COLLECT; the spin date was considered to be day one rather than day zero, as the larva could have spun its cocoon anytime in the 24 hours since the previous day's inspection, therefore specimens age "1" were collected on the morning that their cocoon was first present. Storage proteins are HEX1, hexamerin 1; HEX2, hexamerin 2; HiGLX, high glutamine/glutamic acid protein; VHDL, very high density lipoprotein; values are μg/μl of hemolymph; each value is the mean of two or, for most specimens, four determinations.