DOI: 10.5061/dryad.z8w9ghxmb
title: Nutrition affects larval survival and the development of morphological traits in male and female flour beetles, but genital size and shape remain canalized

description: The caloric content and macronutrient ratio of the diet consumed is a
 major source of phenotypic variation in most animal populations. While
 these nutritional effects have been well-documented for a variety of
 life-history and morphological traits, the effects of nutrition on male
 genitals are poorly understood but genitals are thought to be more
 canalised than general morphology and hence less susceptible to variation
 in nutrition. Even less is known about the effects of nutrition on female
 genital form, which to our knowledge, have never been investigated. Here
 we tested for the effects of juvenile dietary macronutrients (protein and
 carbohydrate) on larval survival, and adult morphology, including genital
 size and shape in male and female flour beetles (Tribolium castaneum). We
 found there was nutritionally induced plasticity in larval survival and
 morphology, although the latter effect was variable, with body size being
 most responsive to dietary macronutrients and genital size and shape being
 least responsive. Functionally equivalent morphological traits in the
 sexes responded similarly to nutrition. Previously, we showed that the
 genitalia of male and female T. castaneum are subject to strong
 stabilizing sexual selection, and our current findings suggest that
 developmental mechanisms reduce the nutritional sensitivity of male and
 female genitals, possibly to ensure matching during mating.
description: The size of the male and female beetles was a simple, linear
 measurement of body and elytra size. The size and shape (4 relative warps)
 of male and female genitalia were extracted using geometric morphometrics.
 The data was z-score transformed before analysis and the details of the
 analysis are provided in the manuscript. The survival data had a binomial
 distribution, so a binary GLMM fitted in the MCMCglmm package of R
 (version 3.1.2; Hadfield 2010) to test the statistical significance of
 these coefficients. The full details for these analyses are provided in
 the Supplementary Info, Text S2. The genitalia were
 dissected using a dissecting microscope (Leica125, Leica Microsystems
 Ltd., Bletchley, UK), and an image was captured using a digital microscope
 camera (Pixelink M20-CYL). These images are unavailable (the student
 completed her studies in 2016 and has employment elsewhere). The details
 for the morphometrics analysis are provided in the manuscript but we also
 include 1) the links to the SB Morphometrics website (see below) and the
 co-ordinate data for each male and female genital image. To open these
 files (.NTS files), go to the SB Morphometrics website. In
 "Software:" -&gt; "Thin-plate spline" -&gt;
 "Relative warps analysis" -&gt; download the program
 tpsRelw32 program. Within tpsRelw32 open the Data - NGFemAllDiets OR
 NGMaleAllDiets and in 'File' select 'Open sliders
 file' - these are notepads that specify which landmarks are fixed or
 sliding landmarks. Compute the "Consensus", "Partial
 warps" and "Relative warps". Finally, visualize the
 "Consensus" shape and "Relative warps" in the
 "Display"
description: # Nutrition affects larval survival and the development of morphological
 traits in male and female flour beetles, but genital size and shape remain
 canalized
 [https://doi.org/10.5061/dryad.z8w9ghxmb](https://doi.org/10.5061/dryad.z8w9ghxmb) This data package contains 3 Excel files with raw data and 3 .csv files of the same data.  1. Genitals_nutrition_SurvivalData = the survival of larvae (0 = death, 1 = survived) when developing in 1 of 24 diets 2. RawMaleMorphologyData17_06_2024 = the morphology of male offspring that survived to eclosion 3. RawFemaleMorphologyData17_06_2024 = the morphology of female offspring ## Version changes: **12-sept-2024:** we have included the raw cartesian coordinates for the fixed and semi-landmarks for every male (file name = NGMalAllDiets) and every female (file name = NGFemAllDiets) genitalia. We also include a 'slider file' for male (file name = MSliders) and female (file name = fsliders) genitalia which defines which landmarks are fixed and those on the slide. These files are simple text files as this is how the cartesian coordinates are listed once the images have been processed in tpsdig264, (as described in the MS). ## Genitals\_nutrition\_SurvivalData col 1_ID of individual larva in a 24-well plate col 2_diet number (coded from 1 to 24) col 3_percentage protein (P) for the diet col 4_percentage carbohydrate (C) for the diet col 5_there were a total of nine replicate 24-well plates for a diet col 6_response variable = survival. Larva that died were coded as a '0', or '1' if they survived. ## RawFemaleMorphologyData17\_06\_2024 col 1_number of rows of data (redundant column) col 2_unique identifiers for individual beetles (ID) col 3_diet number (coded from 1 to 24) col 4_there were a total of nine replicate 24-well plates for a diet. Not all beetles were measured and survival was very poor on some diets so the number of replicates was variable col 5_percentage protein (P) for the diet col 6_percentage carbohydrate (C) for the diet col 7_response variable - female pronotum width (PW, units = mm) col 8_response variable - female elytra width (EL, units = mm) col 9_response variable - female centroid size (CS = genital size, the square root of landmarks from the centroid, unit-less measurement ) col 10_response variable - female relative warp 1 (RW1 = genital shape, principal components analysis of partial warps, unit-less measurement) col 11_response variable - female relative warp 2 (RW2 = genital shape, principal components analysis of partial warps, unit-less measurement) col 12_response variable - female relative warp 3 (RW3 = genital shape, principal components analysis of partial warps, unit-less measurement) col 13_response variable - female relative warp 4 (RW4 = genital shape, principal components analysis of partial warps, unit-less measurement) ## RawMaleMorphologyData17\_06\_2024 col 1_number of rows of data (redundant column) col 2_unique identifiers for individual beetles (ID) col 3_diet number (coded from 1 to 24) col 4_there were a total of nine replicate 24-well plates for a diet. Not all beetles were measured, and survival was very poor on some diets, so the number of replicates was variable col 5_percentage protein (P) for the diet col 6_percentage carbohydrate (C) for the diet col 7_response variable - male pronotum width (PW, units = mm) col 8_response variable - male elytra width (EL, units = mm) col 9_response variable - male centroid size (CS = genital size, the square root of landmarks from the centroid, unit-less measurement ) col 10_response variable - male relative warp 1 (RW1 = genital shape, principal components analysis of partial warps, unit-less measurement) col 11_response variable - male relative warp 2 (RW2 = genital shape, principal components analysis of partial warps, unit-less measurement) col 12_response variable - male relative warp 3 (RW3 = genital shape, principal components analysis of partial warps, unit-less measurement) col 13_response variable - male relative warp 4 (RW4 = genital shape, principal components analysis of partial warps, unit-less measurement) ## **NGFemAllDiets** This is a text file of raw cartesian coordinates for each female genital image. The first row is the number of landmarks LM=36. This is followed by the x and y coordinate data for each landmark. Each x and y coordinate is on a new row. Row 38 is the name of the image Row 39 is the image ID starting at 0 This format is repeated for the next image. ## NGMalAllDiets This is a text file of coordinate data for male genital images. The first row is the number of landmarks LM=19. This is followed by the x and y coordinate data for each landmark. Each x and y coordinate is on a new row. Row 21 is the name of the image Row 22 is the image ID starting at 0 This format is repeated for the next image. ## fsliders This is a text file that specifies which landmarks are 'sliders' row 1 is the name of the file row 2 describes the attributes of the file - 1 18 3 0 - "1" is the 1st landmark, "18" is the number of sliders, and "3" is the number of columns. The following rows specify that the sliders, slide between to other landmarks. For example, "1" is a fixed landmark, it doesn't slide. "2" slides between "1" and "3". "3" slides between "2" and "4" etc. The fixed landmarks never slide and are only ever in the 1st or 3rd column of data. The final landmark is "36" and is fixed. ## MSliders This is a text file that specifies which landmarks are 'sliders' row 1 is the name of the file row 2 describes the attributes of the file - 1 14 3 0 - "1" is the 1st landmark, "14" is the number of sliders, and "3" is the number of columns. The following rows specify that the sliders, slide between to other landmarks. For example, "1" is a fixed landmark, it doesn't slide. "2" slides between "1" and "3". "3" slides between "2" and "4" etc. The fixed landmarks never slide and are only ever in the 1st or 3rd column of data. The final landmark is "19" and is fixed.