DOI: 10.5061/dryad.h18931zzz
title: Data from: The relationship between morphology and flight in Drosophila: a first approach to its genetic basis

description: Flight is a crucial activity for winged insects, involving diverse
 behaviors, and wing morphology has often been proposed as a key factor
 influencing flight capacity. Traits such as wing loading, wing:ratio, and
 wing aspect ratio have been suggested as targets of natural selection,
 exhibiting environmental and genetic variability. Here, we evaluate the
 relationship between morphological traits and two aspects of flight
 performance: flight duration (PTF) and its robustness (CVPTF) in
 Drosophila melanogaster. Additionally, we included wing
 conformation (i.e., wing shape without the component explained by size),
 using geometric morphometrics. We analyzed whether variation of flight
 capacity can be attributed to morphological variation, employing 53 lines
 of Drosophila melanogaster whose genome is fully sequenced. 
description: For the morphological analysis, 10-20  flies of each sex per line
 were randomly selected. Wings and thoraxes were dissected and mounted in a
 standardized way, then photographed using a binocular microscope.
 Morphological traits were measured from these images using
 <em>tpsDig</em>. For wing traits, 15 landmarks were placed on
 the ventral side of the left wing. Wing shape was estimated using the
 <em>Morpho</em> package, allowing shape to be decomposed into
 size and conformation components.  The images were used
 to obtain also the following lineal measurements: thorax length (TL, the
 distance between the anterior margin of the thorax and the tip of the
 scutellum), total wing length (TWL, the distance between landmarks 6 and
 13; Figure S1), and wing width (WW, the distance between landmarks 12 and
 15; Figure S1). Then, we estimated the compound traits wing loading (WLD),
 wing:thorax ratio (WTR), and wing aspect ratio (WAS): WLD as
 (TL)<sup>3</sup>/(WAR), WTR as WAR/TL , and WAS as TWL/WW.
 Additionally, for each morphological trait, we estimated its coefficient
 of variation at the line level.<br> <br> We estimated flight
 capacity using two different traits: the proportion of time in flight
 (PTF) over two minutes, and its coefficient of environmental variation,
 which represents a measure of its robustness (CVPTF). To evaluate PTF, we
 used a previously tested flight chamber associated with a camera set at a
 1080 x 720 resolution and a frame rate of 30 frames per second.
 All flight capacity measurements were carried out in controlled
 temperature conditions (25±1°C), employing virgin flies of 3–7 days old.
 We defined flight time as the time spent by flies flying above a height of
 five cm inside the chamber. Finally, each recorded video was analyzed
 using specially designed motion-tracking software (https://github.com/galimba/drosophila). Flies that were damaged by the rod or those that could not fly were removed from the analysis. CVPTF was estimated for each sex separately as σ/μ, where μ stands for the PTF line’s mean and σ represents the standard deviation for the corresponding line.
description: # Data from: The relationship between morphology and flight in Drosophila:
 a first approach to its genetic basis Dataset DOI:
 [10.5061/dryad.h18931zzz](10.5061/dryad.h18931zzz) ## Description of the
 data and file structure The 53 DGRP lines used in this study were obtained
 from the Bloomington Drosophila Stock Center and were randomly chosen. For
 each line, 100 sexually mature couples were released into an
 egg-collecting chamber (a plastic container of 15 x 10 x 4 cm) containing
 a Petri dish with egg-laying medium (2% agar in distilled water and
 baker’s yeast). Eggs were allowed to hatch, and batches of 30 first-instar
 larvae were transferred to culture vials containing 5 mL of lab medium.
 Larvae were raised at 25±1°C and 60–70% humidity with a 12:12 light:dark
 photoperiod until adult emergence. For each line, we generated ten
 replicates (vials), with five randomly assigned to morphological
 measurements and the other five to flight-capacity estimates.
 **Morphological measurements** For morphological characterization, we
 randomly chose 4 to 6 flies of each sex per vial. The wings and the thorax
 of each individual were removed and mounted on a flat surface, always
 following the same arrangement. Then, they were photographed using a
 binocular microscope (10x) with an attached digital camera connected to a
 computer. We used the images to measure different morphological traits
 using tpsDig. For the estimation of wing traits, 15 landmarks were
 digitized on the ventral face of the left wing of each fly (Figure S1). We
 estimated wing shape (a multivariate morphological trait) using the Morpho
 package (Schlager, 2017). This methodology allows the separation of shape
 into its components: size and conformation. The images were used to obtain
 also the following lineal measurements: thorax length (TL, the distance
 between the anterior margin of the thorax and the tip of the scutellum),
 total wing length (TWL, the distance between landmarks 6 and 13; Figure
 S1), and wing width (WW, the distance between landmarks 12 and 15; Figure
 S1). Also, we calculated wing area (WAR) as TWL x WW. Then, we estimated
 the compound traits wing loading (WLD), wing:thorax ratio (WTR), and wing
 aspect ratio (WAS): WLD as (TL)3/(WAR) WTR as WAR/TL, and WAS as TWL/WW.
 ### Files and variables #### File: Flight_analysis.R **Description:** Code
 of statistical analysis for flight performance. #### File:
 Morphology_analysis.R **Description:** Code of statistical analysis for
 morphological measurements. #### File: Raw_morpho_data.csv
 **Description:** This data set includes flies randomly chosen of each sex
 per vial (between 4 to 6). The wings and the thorax of each individual
 were removed and mounted on a flat surface, always following the same
 arrangement. Then, they were photographed using a binocular microscope
 (10x) with an attached digital camera connected to a computer. The images
 were used to obtain also the following lineal measurements: thorax length
 (TL, the distance between the anterior margin of the thorax and the tip of
 the scutellum), total wing length (TWL, the distance between landmarks 6
 and 13; Figure S1 in the manuscript), and wing width (WW, the distance
 between landmarks 12 and 15; Figure S1). Then, we estimated the compound
 traits wing loading (WLD), wing:thorax ratio (WTR), and WAS as TWL/WW.
 ##### Variables * LINE:Name or code of the line from the Bloomington
 Drosophila Stock Center. (Qualitative random variable). * REPLICATE:Vial
 from which the fly was bred. (Qualitative random variable). * SEX: Sex of
 the flies (male or female). (Qualitative fixed variable). *
 Wlenght_mm:Total wing length, (TWL) the distance between landmarks 6 and
 13; Figure S1, expressed in mm. * Wwidth_mm:Wing width (WW), the distance
 between landmarks 12 and 15; Figure S1,  expressed in mm. *
 Tlenght_mm:Thorax length (TL), the distance between the anterior margin of
 the thorax and the tip of the scutellum, expressed in mm. * Warea_mm2:we
 calculated wing area (WAR) as TWL x WW, expressed in mm2. * WLD_mm:Wing
 loading was calculated as (TL x TL x TL)/(WAR),  expressed in mm *
 WTR_mm:Wing:thorax Ratio was calculated as WAR/TL,  expressed in mm *
 WAS:Wing aspect Ratio was calculated as TWL/WW. #### File:
 Raw_Flight_data_for_individuals.csv **Description:** This file contains
 the value of time in flight in two minutes of experiment for 1905 flies
 belonging to 53 lines of Drosophila melanogaster of the Bloomington
 Drosophila Stock Center. These lines were were randomly chosen from all
 stock of avalaible lines (aproximately 200). ##### Variables * BLOCK:
 Flight time measurement date. (Qualitative random variable). * LINE:Name
 or code of the line from the Bloomington Drosophila Stock Center.
 (Qualitative random variable). * SEX: Sex of the flies (male or female).
 (Qualitative fixed variable). * TIME_FLIGHT:Flight time in seconds during
 2 minutes of experiment. (Quantitative variable) PTF:Mean proportion of
 flight time for each sexual line. (Quantitative variable). #### File:
 Raw_Flight_data_for_lines.csv **Description:** This file contains the
 value of time in flight in two minutes of experiment for 1905 flies
 belonging to 53 lines of Drosophila melanogaster of the Bloomington
 Drosophila Stock Center. These lines were were randomly chosen from all
 stock of avalaible lines (aproximately 200). ##### Variables * LINE:Name
 or code of the line from the Bloomington Drosophila Stock Center.
 (Qualitative random variable). * SEX:Sex of the flies (male or female).
 (Qualitative fixed variable). * PTF:Mean proportion of flight time for
 each sexual line. (Quantitative variable). * SD:Standard deviation of the
 proportion of flight time for each sexual line. (Quantitative variable). *
 CV:Coefficient of variation (SD/TFP) of the proportion of flight time for
 each combination of line and sex. (Quantitative variable). * BLOCK:Flight
 time measurement date. (Qualitative random variable). * WLD_mm:Wing
 loading was calculated as (TL x TL x TL)/(WAR),  expressed in mm *
 WTR_mm:Wing:thorax Ratio was calculated as WAR/TL,  expressed in mm *
 WAS:Wing aspect Ratio was calculated as TWL/WW. #### File:
 Raw_morphogeo_data.csv **Description:** For the estimation of wing traits,
 15 landmarks were digitized on the ventral face of the left wing of each
 fly (Figure S1 in the manuscript). We estimated wing shape (a multivariate
 morphological trait) using the Morpho package. This methodology allows the
 separation of shape into its components: size and conformation. We
 employed 1077 wing photos, corresponding between 4-6 individuals per sex
 and line combination. All wings were superimposed to minimise differences
 relative to size, position and orientation from Procrustes Analysis. This
 procedure generated 26 new Procrustes coordinates and eliminated four
 degrees of freedom, resulting in 26 shape-space dimensions. Shape
 variables generated afterwards, known as partial warps, indicate the
 partial contributions of hierarchically scaled vectors spanning a linear
 shape-space. Subsequently, principal components analysis of the partial
 warps scores matrix was conducted to obtain 26 new shape variables called
 relative warps (RWs). Hence, the dataset includes 26 relative warps for
 both sexes of lines of D. melanogaster.\ Relative deformations are
 represented by the principal components. In this case, PC1 to PC26, and
 they dont have units. \ On the other hand, the variables (here represented
 by X.1, Y.1 to X.15 and Y.15 correspond to coordinates of each landmarck
 in the wing. ##### Variables * LINE:Name or code of the line from the
 Bloomington Drosophila Stock Center. (Qualitative random variable). *
 REPLICATE:Vial from which the fly was bred. (Qualitative random variable).
 * SEX:Sex of the flies (male or female). (Qualitative fixed variable). *
 LINE_SEX: qualitative variable combination of LINE and SEX.(Qualitative
 random variable). * Centroide_Size:Centroid size was calculated as the
 square root of the sum of the squared distances from each landmark to the
 centroid (geometric center) of the configuration of landmarks. The units
 are pixels. * PC1:This column correspond to Relative warps. The response
 variables are the relative warps, and them do not have unities. * PC2:This
 column correspond to Relative warps. The response variables are the
 relative warps, and them do not have unities. * PC3:This column correspond
 to Relative warps. The response variables are the relative warps, and them
 do not have unities. * PC4:This column correspond to Relative warps. The
 response variables are the relative warps, and them do not have unities. *
 PC5:This column correspond to Relative warps. The response variables are
 the relative warps, and them do not have unities. * PC6:This column
 correspond to Relative warps. The response variables are the relative
 warps, and them do not have unities. * PC7:This column correspond to
 Relative warps. The response variables are the relative warps, and them do
 not have unities. * PC8:This column correspond to Relative warps. The
 response variables are the relative warps, and them do not have unities. *
 PC9:This column correspond to Relative warps. The response variables are
 the relative warps, and them do not have unities. * PC10:This column
 correspond to Relative warps. The response variables are the relative
 warps, and them do not have unities. * PC11:This column correspond to
 Relative warps. The response variables are the relative warps, and them do
 not have unities. * PC12:This column correspond to Relative warps. The
 response variables are the relative warps, and them do not have unities. *
 PC13:This column correspond to Relative warps. The response variables are
 the relative warps, and them do not have unities. * PC14:This column
 correspond to Relative warps. The response variables are the relative
 warps, and them do not have unities. * PC15:This column correspond to
 Relative warps. The response variables are the relative warps, and them do
 not have unities. * PC16:This column correspond to Relative warps. The
 response variables are the relative warps, and them do not have unities. *
 PC17:This column correspond to Relative warps. The response variables are
 the relative warps, and them do not have unities. * PC18:This column
 correspond to Relative warps. The response variables are the relative
 warps, and them do not have unities. * PC19:This column correspond to
 Relative warps. The response variables are the relative warps, and them do
 not have unities. * PC20:This column correspond to Relative warps. The
 response variables are the relative warps, and them do not have unities. *
 PC21:This column correspond to Relative warps. The response variables are
 the relative warps, and them do not have unities. * PC22:This column
 correspond to Relative warps. The response variables are the relative
 warps, and them do not have unities. * PC23:This column correspond to
 Relative warps. The response variables are the relative warps, and them do
 not have unities. * PC24:This column correspond to Relative warps. The
 response variables are the relative warps, and them do not have unities. *
 PC25:This column correspond to Relative warps. The response variables are
 the relative warps, and them do not have unities. * PC26:This column
 correspond to Relative warps. The response variables are the relative
 warps, and them do not have unities. * 1.X:This column correspond to
 coordinates of each landmarck in the wing respect the X or Y axis. *
 1.Y:This column correspond to coordinates of each landmarck in the wing
 respect the X or Y axis. * 2.X:This column correspond to coordinates of
 each landmarck in the wing respect the X or Y axis. * 2.Y:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 3.X:This column correspond to coordinates of each landmarck in the
 wing respect the X or Y axis. * 3.Y:This column correspond to coordinates
 of each landmarck in the wing respect the X or Y axis. * 4.X:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 4.Y:This column correspond to coordinates of each landmarck in the
 wing respect the X or Y axis. * 5.X:This column correspond to coordinates
 of each landmarck in the wing respect the X or Y axis. * 5.Y:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 6.X:This column correspond to coordinates of each landmarck in the
 wing respect the X or Y axis. * 6.Y:This column correspond to coordinates
 of each landmarck in the wing respect the X or Y axis. * 7.X:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 7.Y:This column correspond to coordinates of each landmarck in the
 wing respect the X or Y axis. * 8.X:This column correspond to coordinates
 of each landmarck in the wing respect the X or Y axis. * 8.Y:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 9.X:This column correspond to coordinates of each landmarck in the
 wing respect the X or Y axis. * 9.Y:This column correspond to coordinates
 of each landmarck in the wing respect the X or Y axis. * 10.X:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 10.Y:This column correspond to coordinates of each landmarck in
 the wing respect the X or Y axis. * 11.X:This column correspond to
 coordinates of each landmarck in the wing respect the X or Y axis. *
 11.Y:This column correspond to coordinates of each landmarck in the wing
 respect the X or Y axis. * 12.X:This column correspond to coordinates of
 each landmarck in the wing respect the X or Y axis. * 12.Y:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 13.X:This column correspond to coordinates of each landmarck in
 the wing respect the X or Y axis. * 13.Y:This column correspond to
 coordinates of each landmarck in the wing respect the X or Y axis. *
 14.X:This column correspond to coordinates of each landmarck in the wing
 respect the X or Y axis. * 14.Y:This column correspond to coordinates of
 each landmarck in the wing respect the X or Y axis. * 15.X:This column
 correspond to coordinates of each landmarck in the wing respect the X or Y
 axis. * 15.Y:This column correspond to coordinates of each landmarck in
 the wing respect the X or Y axis. #### Raw Data * `LANDMARCKS53LINES.TPS `
 This file consists of the position of each reference point used to study
 the conformation and size of the wing. * `Video_Example.MOV` This is a
 video recorded associated with a flight of one fly included in this study.
 * `Wings_and_Thoraxes_photos.zip` This compressed file contains all the
 photos of the wings and thoraces used in this study.\ \ This file contains
 all the wing and thorax photos used in this study in two different
 folders. The name of each photo follows this code, where the underscore
 separates the other parts of the code. For example, 38_A_H_THORAX_01_02
 The first number corresponds to the name of the isogenic line (in this
 case, 38). The first letter corresponds to the name of the replicate, the
 vial where the flies were stored (in this case, A). The next part of the
 code is a dichotomous coding that refers to the sex of the fly, where H
 corresponds to female and M to male. The next part of the code is a
 dichotomous coding that refers to the morphological trait of the fly
 (THORAX or WING). Here, these terms are written in Spanish, so ALA
 corresponds to WING and THORAX to THORAX. The next number, in this case
 01, corresponds to the fly's identifier. It is possible that, in the
 case of a particular fly, the wings or thorax were damaged during the
 mounting of the specimen under the dissection magnifying glass, so it is
 possible that only one photo of the fly was available. Finally, the last
 number (in this case, 02) corresponds to the photo number of this
 specimen. This last part of the code is irrelevant. In addition, this file
 includes two photos associated with the scale used. All photos were taken
 at the same magnification. Each square in the photo corresponds to 1
 millimeter. ## Code/software All statistical analyses were performed in R
 version 4.3.3 (R Core Team, 2024).\ Here we provide the script for the
 analysis associated with flight and morphological data.