Resolution of three cryptic agricultural pests (Ceratitis fasciventris, C. anonae, C. rosa, Diptera: Tephritidae) using cuticular hydrocarbon profiling. Lucie Vaníčková a,b, * , Massimiliano Virgilio c , Aleš Tomčala b , Radka Břízová b,d , Sunday Ekesi e , Michal Hoskovec b , Blanka Kalinová b , Ruth Rufino Do Nascimento a , Marc De Meyer c a Institute of Chemistry and Biotechnology, Federal University of Alagoas, BR 104 Norte Km 14, 57072-970 Maceió, Alagoas, Brazil b Institute of Organic Chemistry and Biochemistry of the ASCR, Flemingovo nám. 2, CZ-166 10 Prague 6, Czech Republic c Royal Museum for Central Africa, Leuvensesteenweg 13, B-3080 Tervuren, Belgium d Institute of Chemical Technology in Prague, Technická 5, CZ-166 28 Prague 6, Czech Republic e International Centre of Insect Physiology and Ecology, PO Box 30772-00100 GPO, Nairobi, Kenya. * Corresponding author E-mail address: [email protected](L. Vaníčková)
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Resolution of three cryptic agricultural pests (Ceratitis fasciventris, C. anonae, C. rosa,
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Tables
Table 1
PERMANOVA and a posteriori comparisons (t statistic) testing the differences in the multivariate patterns of 59 CHCs in response to the species (C. fasciventris, C. anonae, C. rosa, C. capitata) and the sex (male vs. female) of 80 tephritid fruit flies. The P-values were obtained using 105 unrestricted permutations of raw data. d.f.: degrees of freedom; MS: mean-square estimates; F: pseudo-F. Probability of Monte Carlo simulations: n.s.: not significant, at P<0.05; ***: P<0.001, **: P<0.01; *: P<0.05.
Source of
variability d.f. MS F
Species 3 12650.38 594.54 ***
Sex 1 1595.88 1.103 n.s.
Species × sex 3 1445.72 67.94 ***
Residual 72 21.27
Euclidean distances, log-transformed data
Pairwise a posteriori comparisons:
Species P level
C. capitata vs. C. fasciventris ***
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C. capitata vs. C. anonae ***
C. capitata vs. C. rosa ***
C. fasciventris vs. C. anonae ***
C. fasciventris vs. C. rosa ***
C. rosa vs. C. anonae ***
Species × sex P level
C. capitata: male vs. female ***
C. fasciventris: male vs. female ***
C. anonae: male vs. female *
C. rosa: male vs. female ***
Table 2The compounds identified by chemical (GC×GC/TOFMS) and statistical (SIMPER and PERMANOVA) analyses in the hexane body-washes of females and males of Ceratitis fasciventris, C. anonae, C. rosa and C. capitata. n=10 for each combination of gender and species. RI: retention index; * : present, - : absent.
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Suppl. Table 1 CHC dissimilarity between species. The CHC retention index (RI), (log-transformed data, SD in parentheses), Bray-Curtis dissimilarity (i), the consistency of the contribution to i across specimens (i/SD(i)) and the percentage contribution to the average dissimilarity as calculated by SIMPER (Clarke, 1993). For each pairwise comparison, the table shows the first ten CHCs (out of the 59 peaks) that contribute the most to the dissimilarity. The asterisks indicate the compounds identified in Table 2 and Fig. 2.
% contrib. to
RI C. anonae C. fasciventris i i /SD(i) dissimilarity
2978* 15.21 (0.60) - 0.94 27.46 10.15
2967* 14.90 (0.40) - 0.92 36.72 9.94
3529* 13.60 (0.74) - 0.84 19.29 9.07
2914* - 13.57 (1.23) 0.84 10.93 9.05
3488* - 13.17 (0.16) 0.81 57.53 8.79
3277* - 12.72 (0.51) 0.78 26.5 8.48
3077* 15.84 (1.19) 7.87 (8.09) 0.52 1.14 5.63
2877* 15.52 (1.49) 12.24 (0.45) 0.2 2.17 2.19
3267 15.15 (0.50) 15.25 (0.33) 0.15 3.78 1.67
3770 16.30 (1.20) 14.42 (1.28) 0.13 1.41 1.39
average dissimilarity = 9.24
RI C. fasciventris C. rosa
2967* - 14.58 (0.36) 0.89 38.83 10.71
3630* - 14.52 (0.46) 0.89 32.66 10.67
2978* - 14.00 (0.35) 0.86 38.88 10.29
3529* - 13.62 (0.24) 0.83 54.86 10.01
2914* 13.57 (1.23) - 0.83 10.95 9.97
3077* 7.87 (8.09) - 0.48 1.00 5.80
2877* 12.24 (0.45) 6.13 (6.29) 0.39 1.06 4.64
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3770 14.42 (1.28) 16.98 (0.40) 0.16 1.93 1.89
3256 13.48 (1.14) 15.91 (0.50) 0.15 2.01 1.79
3267 15.25 (0.33) 17.47 (0.79) 0.14 2.72 1.64
average dissimilarity = 8.32
RI C. anonae C. rosa
3077* 15.84 (1.19) - 0.94 14.08 14.39
3277* - 14.52 (0.73) 0.86 19.48 13.20
3635* - 14.52 (0.46) 0.86 33.12 13.20
3488* - 13.30 (0.68) 0.79 20.48 12.09
2877* 15.52 (1.49) 6.13 (6.29) 0.56 1.49 8.53
3780 14.93 (1.78) 16.02 (1.02) 0.11 1.28 1.64
3046 13.42 (0.66) 15.19 (0.38) 0.10 2.37 1.60
3489 17.79 (1.10) 16.70 (1.19) 0.09 1.36 1.41
3790 14.25 (1.19) 15.01 (0.96) 0.08 1.26 1.20
average dissimilarity = 6.54
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Suppl. Table 2 CHC dissimilarity between the sexes within each species. The CHC retention index (RI) (log-transformed data, SD in parentheses), Bray-Curtis dissimilarity (i), the consistency of the contribution to i across specimens (i /SD( i)) and the percentage contribution to the average dissimilarity as calculated by SIMPER (Clarke, 1993). For each pairwise comparison, the table shows the first ten CHCs (out of 59 peaks) that contribute the most to the dissimilarity. The asterisks indicate the compounds identified in Table 2.
C. fasciventris % contrib. to
RI male female i i /SD(i) dissimilarity
3077* - 15.73 (0.85) 1.00 20.71 25.05
3770 15.26 (0.64) 13.58 (1.22) 0.11 1.48 2.82
3760 13.91 (0.38) 13.50 (0.60) 0.11 1.4 2.67
2914* 12.89 (0.32) 14.24 (1.44) 0.10 1.45 2.56
3448 17.00 (0.36) 15.79 (1.79) 0.10 1.02 2.42
3357 14.06 (0.10) 12.63 (1.26) 0.09 1.31 2.36
3489 16.87 (0.25) 16.46 (1.56) 0.09 2.26 2.28
3256 13.82 (1.23) 13.15 (0.99) 0.09 1.72 2.26
3277* 15.56 (0.20) 14.21 (0.99) 0.09 1.53 2.21
3236 16.67 (0.10) 16.03 (1.47) 0.08 1.38 1.99
average dissimilarity = 3.99
C. anonae
RI male female
3780 14.93 (1.83) 14.83 (1.83) 0.11 1.21 4.31
2877* 15.63 (1.47) 15.41 (1.58) 0.09 1.16 3.53
3077* 15.10 (1.30) 16.57 (0.29) 0.09 1.26 3.46
3448 17.02 (1.26) 16.61 (1.50) 0.08 0.97 3.01
3498 17.72 (1.40) 17.73 (0.70) 0.08 1.72 2.92
3377 14.98 (0.39) 13.85 (1.28) 0.08 1.12 2.90
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3489 17.59 (0.99) 17.99 (1.22) 0.08 1.43 2.87
3770 16.30 (1.23) 16.30 (1.24) 0.08 1.18 2.87
3528 16.25 (1.00) 15.62 (1.06) 0.07 1.42 2.82
3790 14.25 (1.23) 14.25 (1.23) 0.07 1.12 2.79
average dissimilarity = 2.65
C. rosa
RI male female
2877* - 12.27 (0.18) 0.72 76.64 28.05
2886 13.75 (0.78) 14.11 (0.62) 0.05 1.26 1.81
3267 17.32 (1.00) 17.62 (0.50) 0.05 1.00 1.79
3277* 14.60 (0.63) 14.44 (0.85) 0.05 1.27 1.83
3458 17.12 (0.43) 16.34 (1.17) 0.06 0.93 2.21
3489 16.84 (0.95) 16.55 (1.43) 0.07 1.17 2.88
3498 17.13 (1.29) 16.95 (0.75) 0.07 1.37 2.64
3528 15.40 (0.27) 14.47 (0.38) 0.05 2.07 2.12
3780 16.60 (0.56) 15.43 (1.07) 0.08 1.34 2.99
3790 15.57 (0.89) 14.44 (0.67) 0.08 1.91 3.14
average dissimilarity = 2.58
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Figure Captions
Fig. 1. The principal component analyses (PCA) of the Euclidean distances (as calculated from the peak areas of 59 CHCs) among the male and female specimens of C. capitata, C. fasciventris, C. anonae and C. rosa.
Fig. 2. The log-transformed peak areas of CHCs (the standard deviations as error bars), represented by their retention indices, those contribute the most to the differences (2% contribution to species dissimilarity) between C. fasciventris, C. anonae and C. rosa (see Suppl. Tab. 1).
Fig. 3. The Principal component analyses (PCA) of the Euclidean distances among the individual genotypes of the specimens sampled in this study (grouped according to the species and the sex, represented in black) and 621 genotypes of C. fasciventris, C. anonae and C. rosa assigned to 5 genotypic clusters (C. fasciventris F1 and F2, C. anonae A, C. rosa R1 and R2) like in Virgilio et al. (2013). The genotype groups are labeled inside their 95% inertia ellipses and connected to the corresponding group centroids.