Title
Cellular basis of morphological variation and temperature-related plasticity in Drosophila melanogaster strains with divergent wing shapes
Journal Name
Genetica
Authors
Libéria Souza Torquato; Daniel Mattos; Bruna Palma Matta; Blanche Christine Bitner-Mathé
Affiliations
Laboratório de Evolução de Caracteres Complexos – Drosophila, Departamento de Genética, Instituto de Biologia, Universidade Federal do Rio de Janeiro – Brazil
Corresponding author
Blanche Christine Bitner-Mathé
email: bcbitner@biologia.ufrj.br/ bcbitner@gmail.com
Mailing address: Avenida Carlos Chagas Filho, 373, Sala A2-109, Bloco A, Edifício do Centro de Ciências da Saúde, 21941 902, Ilha do Fundão, Cidade Universitária, Rio de
Janeiro, RJ
Phone: +55 21 3938-6387 / +55 21 99559-1970
Fax: +55 21 3938-6333
Running title
Cellular basis of wing shape and size variation
Supplementary Material
Fig. S1: Original wings images superposed with respective estimated ellipses, using actual data from ellipse adjustment to semi-landmarks (traced blue lines) and its proxy via
wing width (WW) and length (WL) measurements (red solid lines). Presented wings were randomly chosen: first specimen in alphabetical order for each D. melanogaster
selection strain (1L, 1R, 5L, 5R). Ellipse adjustment was performed using a least square procedure to solve the ellipse equation for Cartesian coordinates of 20 semi-landmarks
taken on the wing contour (details in Klackzo and Bitner-Mathé 1990; and Klaczko 2006; see main text). In all four cases, correlation between x-observed and their y-expected
ellipse values was greater than 99%. Given that no scaling transformation is applied, the estimated ellipses were reconstructed from values of minor (b) and major (a) ellipse
radii (in mm), each multiplied by 2. In turn, ellipses for the proxy of such adjustment were reconstructed using WW and WL values (in mm). On average, WW estimates 94.5%
of the respective 2b value, while WL estimates 98.7% of the respective 2a value; similar values were found in data from other studies (not shown).
Table S1: Mean ± SE for wing morphology and cellular variations at intervein regions (Fig. 1a) by individual strain and temperature of development (TD).
TD 25°C (mean ± SE)
TD 16°C (mean ± SE)
TD 25°C (mean ± SE)
TD 16°C (mean ± SE)
Trait
1L (n=10)
1R (n=10)
1L (n=10)
1R (n=10)
5L (n=10)
5R (n=10)
5L (n=10)
5R (n=10)
0.445 ± 0.002
0.542 ± 0.001
0.438 ± 0.003
0.564 ± 0.003
0.453 ± 0.002
0.528 ± 0.004
0.439 ± 0.002
0.516 ± 0.004
WSH
0.678 ± 0.003
0.666 ± 0.007
0.745 ± 0.006
0.760 ± 0.005
0.656 ± 0.003
0.684 ± 0.006
0.734 ± 0.011
0.760 ± 0.006
WSI
0.904 ± 0.005
0.980 ± 0.010
0.986 ± 0.011
1.141 ± 0.009
0.883 ± 0.005
0.994 ± 0.008
0.973 ± 0.015
1.092 ± 0.009
WW
2.033 ± 0.007
1.809 ± 0.020
2.251 ± 0.017
2.023 ± 0.011
1.949 ± 0.011
1.885 ± 0.021
2.217 ± 0.035
2.118 ± 0.017
WL
0.129 ± 0.004
0.140 ± 0.003
0.162 ± 0.008
0.184 ± 0.004
0.128 ± 0.002
0.156 ± 0.003
0.154 ± 0.007
0.188 ± 0.007
IVRA
0.250 ± 0.004
0.240 ± 0.005
0.289 ± 0.008
0.317 ± 0.004
0.227 ± 0.004
0.262 ± 0.004
0.285 ± 0.009
0.312 ± 0.007
IVRB
0.280
±
0.004
0.259
±
0.006
0.348
±
0.008
0.348
±
0.005
0.274
±
0.004
0.273
±
0.006
0.347
±
0.012
0.338 ± 0.005
IVRC
0.225 ± 0.003
0.210 ± 0.005
0.280 ± 0.006
0.280 ± 0.003
0.216 ± 0.004
0.227 ± 0.005
0.280 ± 0.009
0.296 ± 0.004
IVRD
0.371 ± 0.004
0.373 ± 0.009
0.468 ± 0.011
0.473 ± 0.008
0.359 ± 0.003
0.371 ± 0.007
0.461 ± 0.018
0.465 ± 0.007
IVRE
2.084 ± 0.049
1.939 ± 0.073
2.207 ± 0.044
2.234 ± 0.059
2.010 ± 0.036
2.002 ± 0.049
2.158 ± 0.063
2.270 ± 0.067
CAA
1.796 ± 0.035
1.687 ± 0.048
1.901 ± 0.053
2.085 ± 0.079
1.751 ± 0.031
1.710 ± 0.027
2.150 ± 0.046
2.044 ± 0.043
CAB
1.735 ± 0.043
1.630 ± 0.071
1.851 ± 0.036
1.846 ± 0.058
1.773 ± 0.036
1.848 ± 0.058
1.945 ± 0.071
1.892 ± 0.081
CAC
1.772 ± 0.042
1.618 ± 0.046
1.901 ± 0.067
1.941 ± 0.045
1.690 ± 0.025
1.691 ± 0.038
1.909 ± 0.035
1.908 ± 0.043
CAD
1.869 ± 0.080
1.609 ± 0.054
2.020 ± 0.081
1.969 ± 0.092
1.838 ± 0.049
1.685 ± 0.054
2.101 ± 0.073
2.004 ± 0.075
CAE
1.851 ± 0.025
1.697 ± 0.046
1.976 ± 0.037
2.015 ± 0.045
1.813 ± 0.021
1.787 ± 0.032
2.053 ± 0.025
2.024 ± 0.030
CAAverage
623.5 ± 28.1
728.7 ± 30.7
736.3 ± 33.1
827.8 ± 28.7
636.7 ± 15.0
780.2 ± 16.4
725.0 ± 48.1
835.5 ± 39.0
CNA
1,393.9 ± 16.6
1,427.1 ± 40.3
1,529.5 ± 47.3
1,541.3 ± 61.1
1,296.1 ± 23.8
1,537.3 ± 42.5
1,326.3 ± 42.8
1,528.6 ± 40.0
CNB
1,621.1 ± 45.4
1,603.8 ± 54.1
1,882.2 ± 51.1
1,899.5 ± 47.0
1,553.5 ± 38.7
1,486.2 ± 49.3
1,795.9 ± 67.0
1,822.1 ± 93.3
CNC
1,258.6 ± 25.5
1,300.9 ± 29.7
1,488.4 ± 48.0
1,456.5 ± 26.8
1,280.8 ± 24.1
1,344.4 ± 33.2
1,478.1 ± 34.6
1,560.3 ± 38.6
CND
2,024.1 ± 105.9
2,352.6 ± 97.7
2,341.5 ± 94.3
2,481.4 ± 115.4
1,963.2 ± 54.8
2,217.3 ± 57.8
2,237.0 ± 124.3
2,346.0 ± 87.9
CNE
7,811.0 ± 102.7 8,243.3 ± 194.4
8,841.1 ± 166.9
9,029.2 ± 163.9
7,467.8 ± 89.8 8,245.4 ± 127.3
8,273.4 ± 238.9 8,997.6 ± 196.6
CNTotal
Wing traits: outline shape (WSH), size (WSI), width (WW), length (WL), and the area of intervein regions A-E (IVRA-E). Cellular traits: average cell area (CA) or cell number (CN)
at intervein regions A-E (subscripts), plus the average cell area across all intervein regions (CAAverage) and total cell number in dorsal wing blade (CNTotal). Units: WSH is a ratio
(WW/WL); WSI, WW and WL are in millimeters, IVR areas are in mm2; and all CA estimates are in mm2 ×10-4.
Table S2: Mean ± SE for wing morphology and cellular variations at
intervein regions in baseline population.
Baseline population
Trait
Wild (n=23)
G1 (n=20)
0.485 ± 0.002
0.471 ± 0.003
WSH
0.632 ± 0.008
0.782 ± 0.006
WSI
0.881 ± 0.012
1.073 ± 0.010
WW
1.815 ± 0.025
2.281 ± 0.018
WL
0.125 ± 0.004
0.184 ± 0.004
IVRA
0.226 ± 0.006
0.329 ± 0.006
IVRB
0.251 ± 0.006
0.385 ± 0.006
IVRC
0.197 ± 0.005
0.309 ± 0.005
IVRD
0.322 ± 0.010
0.505 ± 0.010
IVRE
1.989 ± 0.027
2.349 ± 0.050
CAA
1.797 ± 0.035
2.303 ± 0.055
CAB
1.685 ± 0.044
2.068 ± 0.035
CAC
1.700 ± 0.032
2.087 ± 0.031
CAD
1.816 ± 0.039
2.358 ± 0.062
CAE
1.798 ± 0.025
2.233 ± 0.024
CAAverage
631.4 ± 19.8
790.3 ± 22.0
CNA
1,260.6 ± 32.0
1,436.0 ± 33.3
CNB
1,506.8 ± 43.8
1,873.5 ± 46.4
CNC
1,166.3 ± 31.8
1,489.7 ± 34.1
CND
1,777.9 ± 46.6
2,173.2 ± 72.8
CNE
7,008.0 ± 148.1
8,638.8 ± 164.9
CNTotal
Abbreviations are the same presented in Table S1. G1 flies were reared at
16°C.
TABLE S3: Quadratic regressions of average cell area across rank order of all five intervein regions fitted to complete data set of each
experimental group, and F-tests of group differences from quadratic regressions adjusted to each individual wing.
Experimental
group
n
quadratic regression fitted to complete data set of
each group
g2
MV
IRV-MV
F-ratio
R2
Mean ± SE from quadratic regressions adjusted to
each individual wing
g2
MV
IRV-MV
WILD
23
0.053
1.682
3.417
23.167***
28.0%
0.053 ± 0.009
1.669 ± 0.035
3.588 ± 0.257
G1
20
0.063
2.105
3.156
11.992***
18.2%
0.063 ± 0.013
2.092 ± 0.040
2.944 ± 0.222
L_TD16
20
0.054
1.900
3.337
14.876***
21.9%
0.054 ± 0.011
1.934 ± 0.059
2.556 ± 0.713
R_TD16
20
0.053
1.891
3.627
20.260***
28.0%
0.053 ± 0.015
1.942 ± 0.053
2.708 ± 0.354
L_TD25
20
0.056
1.711
3.382
30.752***
37.5%
0.056 ± 0.010
1.742 ± 0.063
2.261 ± 1.420
R_TD25
20
0.029
1.643
4.192
18.451***
26.1%
0.029 ± 0.009
1.680 ± 0.042
3.018 ± 0.455
F-ratio of group
differencesa
1.067
12.289***
0.459
Abbreviations: coefficient that describes the shape of the curve (g2) and characteristic estimates of minimum value (MV) and intervein region that
holds the minimum value (IVR-MV). A rank order was attributed to each intervein region according to their proximity from the most anterior
region of the wing: IVRA was ranked as 1, IVRB as 2 and so forth. See Fig. 4 for visual representation of the quadratic curves fitted to complete
data set of each experimental group. ANOVA (F-ratio) of group differences was performed in data derived from quadratic regressions adjusted to
each individual wing: group differences (df=5), model error: df=112. aTukeys’s a posteriori pairwise comparisons for the significant group
differences in MV; six outlier cases were excluded: (Wild = R_TD25 = L_TD25) ≠ (G1 = L_TD16 = R_TD16); α = 0.05. G1 flies were reared at
16°C. ***P ≤ 0.001.