ESM2 for the following paper in Ichthyological Research

Title: Interspecific variation in the spawning time of labrid fish on a fringing reef at Iriomote Island, Okinawa

Authors: Tetsuo Kuwamura, Shohei Suzuki, Tatsuru Kadota

Corresponding author: T. Kuwamura, School of International Liberal Studies, Chukyo University, Yagoto-honmachi, Nagoya 466-8666, Japan; e-mail:

Tables S1, S2 and S3

Spawning sites and male mating tactics

Spawning was observed in 30 species of wrasse (Table S1). Eleven species spawned only on the reef edge, nine species spawned only on the patch reef, and the remaining 10 species spawned on both sites (Table S1). Spawning occurred more frequently on the reef edge than on the patch reef (Paired t-test, n = 30, P < 0.05). Gomphosus varius was observed to spawn most frequently (n = 843, mostly on the reef edge), followed by Thalassoma jansenii (n = 479, only on the reef edge).

Pair spawning by territorial TP males was observed in all 30 species (Table S1). Streaking by IP males (and infrequently by TP males) also occurred in eight species, whose spawnings were observed relatively frequently (n > 60). Group spawning by IP males (and rarely with TP males) occurred only in six species, whose spawnings were observed frequently (n > 115), but it did not occur in the frequently observed G. varius, Labrichthys unilineatus, or Epibulus insidiator. The number of streakings positively correlated with the total number of spawnings (r = 0.81, n = 30, P < 0.0001), but that of group spawnings did not significantly correlate with total spawnings (r = 0.26, n = 30, P = 0.16).


Table S1 Spawning sites and male mating tactics of each labrid species. Number of spawns observed at each site and those of each mating tactic are given for each species, arranged in order of observed frequencies

Species Spawning site Male mating tacticsa

reef edge patch reef pair streak group

Gomphosus varius 829 14 829 14b 0

Thalassoma jansenii 479 0 470 7 2c

Thalassoma quinquevittatum 434 0 364d 12 58e

Thalassoma hardwicke 372 2 293f 18g 63h

Labrichthys unilineatus 232 53 285 0 0

Epibulus insidiator 2 210 210 2i 0

Thalassoma amblycephalum 167 0 28 0 139j

Thalassoma lutescens 142 0 129 8 5k

Halichoeres marginatus 114 1 104l 5 6m

Cheilinus trilobatus 93 9 102 0 0

Halichoeres melanurus 62 0 62 0 0

Stethojulis trilineata 60 0 59n 1o 0

Stethojulis bandanensis 53 0 53 0 0

Thalassoma lunare 33 13 46 0 0

Hemigymnus fasciatus 26 3 29 0 0

Cheilinus chlorourus 28 0 28 0 0

Pseudocheilinus hexataenia 19 0 19 0 0

Labroides dimidiatus 16 0 16 0 0

Hemigymnus melapterus 0 12 12 0 0

Oxycheilinus unifasciatus 9 1 10 0 0

Cheilinus oxycephalus 9 0 9 0 0

Bodianus axillaris 1 7 8 0 0

Cheilinus fasciatus 0 7 7 0 0

Labroides bicolor 0 6 6 0 0

Labropsis manabei 0 5 5 0 0

Anampses twistii 0 4 4 0 0

Coris aygula 0 1 1 0 0

Coris gaimard 0 1 1 0 0

Halichoeres melanochir 0 1 1 0 0

Hologymnosus annulatus 0 1 1 0 0

a Pair spawning (pair: a pair only) was usually conducted by TP males, streaking (streak: a pair and one streaker) and group spawning (group: including more than two males) by IP males, and exceptional cases are given below

b 4 by TP, c 1 including TP (streaked by 2 IP), d 1 by IP, e 6 including TP, f 4 by IP, g 1 by TP, h 6 including TP, I 2 by TP, j 6 including TP, k 2 including TP, l 1 by IP, m 1 including TP, n 1 by IP, o by IP into IP pair


Table S2 Results of statistical analyses of the correlation between spawning time and lunar age in each species. The correlation was examined in the period from new moon to full moon (lunar age: 0 to 15 days) and in the period from full moon to new moon (lunar age: 15 to 30 days), respectively. If spawning time shifted from evening to morning (or early afternoon in species that rarely spawned in the morning), it was examined separately in the periods before and after the shift. For each period, lunar age (day), number of spawning events observed (n), and correlation coefficient are given with significance probability

Species (typea) Lunar ageb (n) Correlation coefficient

The first half The second half

E. insidiator (HT) 1.7–16.2 (143) 0.71*** 20.2–30 (62) 0.80***

C. chlorourus (HT) 3.7–15.2 (12) 0.86*** 18.2–30 (11) 0.99***

C. trilobatus (HT) 0–15.2 (74) 0.84*** 20.2–30 (28) 0.93***

H. fasciatus (HT) 0–14.4 (6) 0.90* 19.4–30 (23) 0.78***

T. amblycephalum (HT) 0–16.2 (63) 0.64*** 18.2–24.4 (101) 0.78***

T. jansenii (HT) 0–12.7 (186) 0.80*** 13.2–28.0 (289) 0.57***

T. lunare (HT?) 0–16.2 (27) 0.28 18.2–30 (19) 0.33

T. lutescens (HT) 0–13.2 (60) 0.47*** 14.2–30 (82) 0.73***

T. quinquevittatum (HT) 0–9.7 (127) 0.84*** 13.2–30 (307) 0.35***

G. varius (HT) 0–12.2 (440) 0.75*** 13.2–30 (403) 0.49***

L. dimidiatus (HT) 1.7–12.7 (8) 0.93*** 19.4–27.7 (6) 0.92**

T. hardwicke (HD) 0–13.2 (148) 0.72*** 14.2–30 (226) 0.14*

L. unilineatus (HD) 0–14.7 (139) 0.54*** 15.2–30 (148) 0.56***

S. bandanensis (DA) 2.5–19.2 (33) 0.650** 19.4–30 (17) 0.81***

S. trilineata (DA) 2.7–19.2 (24) 0.66** 21.0–30 (35) 0.85***

H. marginatus (DA) 0–13.2 (41) 0.90*** 14.4–30 (74) 0.22

B. axillaris (DU?) 0–13.4 (4) –0.97* 16.4–30 (4) 0.16

H. melanurus (DU) 0–14.7 (36) –0.30 15.2–30 (26) 0.11

P. hexataenia (DU) 0–11.2 (8) –0.15 17.2–30 (11) 0.14

C. fasciatus (nd) 0–2.3 (1) - 23.7–30 (6) 0.12

C. oxycephalus (nd) 0–12.7 (3) –0.14 17.2–30 (6) 0.40

O. unifasciatus (nd) 0–14.2 (8) –0.27 19.2–30 (2) -

H. melapterus (nd) 0–1.5 (1) - 17.2–30 (11) 0.90***

A. twistii (nd) 0–7.7 (3) 0.82 22.4–30 (1) -

L. manabei (nd) 0–11.2 (2) - 19.2–30 (3) –0.95

L. bicolor (nd) 0–6.7 (3) 1.00* 15.7–30 (3) 0.85

a Type of spawning time: HT, around high tide; HD, high tide and dusk; DA, dusk and afternoon; DU, dusk only; nd, not DU but the type could not be determined owing to small sample size

b Data on days before the shift to morning spawning just after new moon (lunar age 0) and days after the shit to morning spawning just before new moon (lunar age 30) are excluded from the analyses

* P < 0.05, ** P < 0.01, *** P < 0.001


Table S3 Occurrence of dusk spawning in each species. Number of spawning events observed before and after 1756 h (one hour before the earliest sunset time during the study periods: see Table 1) are shown in each 5-day period of lunar age for each species

Species (typea) No. of spawning events before/after 1756 h in each period of lunar age

0–4.9 5–9.9 10–14.9 15–19.9 20–24.9 25–29.9 days

E. insidiator (HT) 39/1 88/0 18/0 4/0 23/0 39/0

C. chlorourus (HT) 6/1 5/0 2/0 3/0 5/0 6/0

C. trilobatus (HT) 37/0 32/0 2/0 0/3 15/0 13/0

H. fasciatus (HT) 5/0 0/0 1/0 2/0 21/0 0/0

T. amblycephalum (HT) 47/0 11/0 3/0 44/1 58/0 3/0

T. jansenii (HT) 84/0 98/0 20/0 51/0 169/0 56/0

T. lunare (HT?) 9/0 10/2 4/0 4/0 6/1 10/0

T. lutescens (HT) 36/0 20/0 5/0 15/0 47/0 19/0

T. quinquevittatum (HT) 70/0 57/0 42/0 45/0 107/0 112/0

G. varius (HT) 239/0 175/0 63/1 74/0 154/0 137/0

L. dimidiatus (HT) 4/0 3/0 3/0 1/0 2/0 3/0

T. hardwicke (HD)b 72/2 47/5 24/7 38/21 93/2 48/15

L. unilineatus (HD)c 23/0 82/0 32/2 56/6 47/0 35/2

S. bandanensis (DA) 7/2 3/1 8/7 5/6 5/0 9/0

S. trilineata (DA) 3/1 2/1 4/3 3/8 14/2 6/13

H. marginatus (DA) 12/0 17/0 3/11 14/10 27/2 5/14

B. axillaris (DU?) 0/0 0/3 1/0 1/2 0/0 0/1

H. melanurus (DU) 0/8 0/12 0/16 0/10 0/6 0/10

P. hexataenia (DU) 0/2 0/4 0/2 0/5 0/2 0/4

C. fasciatus (nd) 1/0 0/0 0/0 0/0 2/0 4/0

C. oxycephalus (nd) 1/0 1/0 1/0 2/1 2/0 1/0

O. unifasciatus (nd) 3/0 2/0 3/0 1/0 1/0 0/0

H. melapterus (nd) 1/0 0/0 0/0 7/0 4/0 0/0

A. twistii (nd) 2/0 1/0 0/0 0/0 1/0 0/0

L. manabei (nd) 1/0 0/0 1/0 2/0 0/0 1/0

L. bicolor (nd) 2/0 1/0 0/0 1/0 1/0 1/0

a Type of spawning time: HT, around high tide; HD, high tide and dusk; DA, dusk and afternoon; DU, dusk only; nd, not DU but the type could not be determined owing to small sample size

b Spawning occurred at dusk in all of the six 5-day periods of lunar age, significantly different from the species of type HT, in which dusk spawning occurred in no or one period (Fisher's exact test, P < 0.01 and 0.05, respectively)

c Spawning occurred at dusk in three of the six 5-day periods of lunar age, but not significantly different from the species of type HT and T. hardwicke (Fisher's exact test, P > 0.05)

1