Interspecific variation in bristle number on forewings of tiny insects does not influence clap-and-fling aerodynamics

Miniature insects must overcome significant viscous resistance in order to fly. They typically possess wings with long bristles on the fringes and use clap-and-fling mechanism to augment lift. These unique solutions to the extreme conditions of flight at tiny sizes (< 2 mm body length) suggest that natural selection has optimized wing design for better aerodynamic performance. However, species vary in wingspan, number of bristles (n), and bristle gap (G) to diameter (D) ratio (G/D). How this variation relates to body length (BL) and its effects on aerodynamics remain unknown. We measured forewing images of 38 species of thrips and 21 species of fairyflies. Our phylogenetic comparative analyses showed that n and wingspan scaled positively and similarly with body length across both groups, whereas G/D decreased with BL, with a sharper decline in thrips. We next measured aerodynamic forces and visualized flow on physical models of bristled wings performing clap-and-fling kinematics at chord-based Reynolds number of 10 using a dynamically scaled robotic platform. We examined the effects of dimensional (G, D, wingspan) and non-dimensional (n, G/D) geometric variables on dimensionless lift and drag. We found that: (a) increasing G reduced drag more than decreasing D; (b) changing n had minimal impact on lift generation; and (c) varying G/D minimally affected aerodynamic forces. These aerodynamic results suggest little pressure to functionally optimize n and G/D. Combined with the scaling relationships between wing variables and BL, much wing variation in tiny flying insects might be best explained by underlying shared growth factors.

Evolution and genetic basis of the plant-penetrating ovipositor, a key adaptation in the transition to herbivory within the Drosophilidae

Herbivorous insects are extraordinarily diverse, yet are found in only one-third of insect orders. This skew may be driven by barriers to plant colonization coupled with phylogenetic constraint on plant-colonizing adaptations. Physical barriers can be surmounted by key innovations like the plant-penetrating ovipositor. Within Drosophilidae, ovipositor margins densely adorned with hard bristles used to cut into plants evolved repeatedly, but their evolutionary, developmental and genomic basis has only been explored in Drosophila suzukii. Here, we addressed this gap using Scaptomyza, an herbivorous radiation nested in a detritivorous clade. First, we found that ovipositor bristle number increased markedly as herbivory evolved in Scaptomyza. We then dissected the genomic architecture of variation in ovipositor bristle number within S. flava using a pooled genome wide association study (pool-GWAS). Variation in ovipositor bristle number in S. flava was heritable and associated with single nucleotide polymorphisms (SNPs) within non-coding regions involved in neural development. Genotyping of individual flies replicated the association at a candidate SNP upstream of Gai, a neural development gene, and estimated that it contributes to an average gain of ∼0.58 bristles/ovipositor in S. flava. Neural developmental genes thus underlie variation in this key morphological adaptation, possibly facilitating the evolution of this trait and the colonization of tough tissue of living plants.

tartan underlies the evolution of Drosophila male genital morphology

Male genital structures are among the most rapidly evolving morphological traits and are often the only features that can distinguish closely related species. This process is thought to be driven by sexual selection and may reinforce species separation. However, while the genetic bases of many phenotypic differences have been identified, we still lack knowledge about the genes underlying evolutionary differences in male genital organs and organ size more generally. The claspers (surstyli) are periphallic structures that play an important role in copulation in insects. Here, we show that divergence in clasper size and bristle number between Drosophila mauritiana and Drosophila simulans is caused by evolutionary changes in tartan (trn), which encodes a transmembrane leucine-rich repeat domain protein that mediates cell–cell interactions and affinity. There are no fixed amino acid differences in trn between D. mauritiana and D. simulans, but differences in the expression of this gene in developing genitalia suggest that cis-regulatory changes in trn underlie the evolution of clasper morphology in these species. Finally, analyses of reciprocal hemizygotes that are genetically identical, except for the species from which the functional allele of trn originates, determined that the trn allele of D. mauritiana specifies larger claspers with more bristles than the allele of D. simulans. Therefore, we have identified a gene underlying evolutionary change in the size of a male genital organ, which will help to better understand not only the rapid diversification of these structures, but also the regulation and evolution of organ size more broadly.

tartan underlies the evolution of male Drosophila genital morphology

Male genital structures are among the most rapidly evolving morphological traits and are often the only features that can distinguish closely related species. This process is thought to be driven by sexual selection and may reinforce species separation. However, while the genetic basis of many phenotypic differences have been identified, we still lack knowledge about the genes underlying evolutionary differences in male genital organs and organ size more generally. The claspers (surstyli) are periphallic structures that play an important role in copulation in insects. Here we show that natural variation in clasper size and bristle number between Drosophila mauritiana and D. simulans is caused by evolutionary changes in tartan (trn), which encodes a transmembrane leucine-rich repeat domain protein that mediates cell-cell interactions and affinity differences. There are no fixed amino acid differences in trn between D. mauritiana and D. simulans but differences in the expression of this gene in developing genitalia suggest cis-regulatory changes in trn underlie the evolution of clasper morphology in these species. Finally, analysis of reciprocal hemizyotes that are genetically identical, except for which species the functional allele of trn is from, determined that the trn allele of D. mauritiana specifies larger claspers with more bristles than the allele of D. simulans. Therefore we have identified the first gene underlying evolutionary change in the size of a male genital organ, which will help to better understand the rapid diversification of these structures and the regulation and evolution of organ size more broadly.Significance StatementThe morphology of male genital organs evolves rapidly driven by sexual selection. However, little is known about the genes underlying genitalia differences between species. Identifying these genes is key to understanding how sexual selection acts on development to produce rapid phenotypic change. We have found that the gene tartan underlies differences between male Drosophila mauritiana and D. simulans in the size and bristle number of the claspers - genital projections that grasp the female during copulation. Moreover, since tartan encodes a protein that is involved in cell affinity, this may represent a new developmental mechanism for morphological change. Therefore, our study provides new insights into genetic and developmental bases for the rapid evolution of male genitalia and organ size more generally.

Exploration of teratogenic and genotoxic effects of fruit ripening retardant Alar (Daminozide) on model organism Drosophila melanogaster

Alar (Daminozide) is a plant growth regulator which is widely used as a fruit preservative for apple and mango to prevent pre-harvest fruit drop, promote color development and to delay excessive ripening. The aim of the present work was to demonstrate the effect of Alar on several life history traits, adult morphology, Hsp70 protein expression and in vivo DNA damage in the brain of the model organism Drosophila melanogaster. We assessed the life history and morphological traits including fecundity, developmental time, pupation height, egg-to-adult viability and mean wing length, body length, arista length and sternopleural bristle number of the emerging flies. The results showed a significant delay in the developmental milestones, increase in body length, wing length, arista length, a decrease in fecundity, pupal height and variation in sternopleural bristle number in the treated flies in comparison to the controls. Overexpression of Hsp70 protein suggests alar induced subcellular molecular stress and comet assay validates genotoxicity in the form of DNA damage in the treated larvae. Mutation screening experiment revealed induction of X lined lethal mutation.

Robustness of bristle number to temperature and genetic background varies with bristle type and is regulated by miR-9a

Robustness is the invariance of a given phenotype when faced with a given incoming genetic or environmental variation. Such essential property is being studied in a wide diversity of traits in many organisms but it is difficult to compare the results obtained on the robustness of different traits with each other given the differences that exist between traits in the way they are measured, in their genetic architecture and development. In this study, we assessed robustness of bristle number to incoming genetic and environmental variation for eight bristle types in the fruit fly Drosophila melanogaster, allowing for a straightforward comparison of the robustness observed between bristle types. We measured the response of bristle number mean and variance to changes in temperature and in the number of copies of two genes (scute and miR-9a) known to be involved in bristle development. Many combinations between the three factors were tested, thus allowing to test for the effect of each factor in different contexts for the two other factors – to which we refer herein as different backgrounds. We have found different responses between bristle types, suggesting that they present different levels of robustness to the factors tested. In addition, we have found that temperature and miR-9a affect more generally the variance of the traits rather than their means, thus fulfilling a criteria usually admitted to identify robustness factors.

The action of evolutionary forces on metric traits

Fisher's theorem of natural selection implies that the population genetic variance of quasi-neutral traits should be mostly additive. In the case of fitness component traits, however, that variance would be characterised by a substantial contribution from non-additive loci. In parallel, Robertson's theorem states that selection will change the population mean of a trait proportionally to the magnitude of the genetic correlation between that trait and fitness, which should be weak for quasi-neutral traits or strong for the mean fitness components. Drosophila data from inbreeding and artificial selection experiments are discussed within that theoretical framework. In addition, the process of regeneration by mutation of the genetic variance of a quasi-neutral trait (abdominal bristle number) in a Drosophila population initially homozygous at all loci has been analysed. After 485 generations of mutation accumulation, the levels of additive variance found in this population closely approached those commonly observed in laboratory populations. Furthermore, these values, together with previously reported estimates for natural populations, could be jointly explained by a model assuming weak causal stabilising selection.

Regeneration of the variance of metric traits by spontaneous mutation in a Drosophila population

SummaryIn the C1 population of Drosophila melanogaster of moderate effective size (≈500), which was genetically invariant in its origin, we studied the regeneration by spontaneous mutation of the genetic variance for two metric traits [abdominal (AB) and sternopleural (ST) bristle number] and that of the concealed mutation load for viability, together with their temporal stability, using alternative selection models based on mutational parameters estimated in the C1 genetic background. During generations 381–485 of mutation accumulation (MA), the additive variances of AB and ST approached the levels observed in standing laboratory populations, fluctuating around their expected equilibrium values under neutrality or under relatively weak causal stabilizing selection. This type of selection was required to simultaneously account for the observed additive variance in our population and for those previously reported in natural and laboratory populations, indicating that most mutations affecting bristle traits would only be subjected to weak selective constraints. Although gene action for bristles was essentially additive, transient situations occurred where inbreeding resulted in a depression of the mean and an increase of the additive variance. This was ascribed to the occasional segregation of mutations of large recessive effects. On the other hand, the observed non-lethal inbreeding depression for viability must be explained by the segregation of alleles of considerable and largely recessive deleterious effects, and the corresponding load concealed in the heterozygous condition was found to be temporally stable, as expected from tighter constraints imposed by natural selection.