bioRxiv Science⌕ Search

Biology subjects

Smee, M. R.

Publications and source records attributed to Smee, M. R..

3 recordsLinked to original sources

Strong GAL4 expression compromises Drosophila fat body function

The ability to direct tissue-specific overexpression of transgenic proteins in genetically tractable organisms like Drosophila melanogaster has facilitated innumerable biological discoveries. However, transgenic proteins can themselves impact cellular and physiological processes in ways that are often ignored or poorly defined. Here we discovered that the yolk-GAL4 transgene, which directs strong expression of the yeast GAL4 transcription factor in the Drosophila fat body, induces significant physiological defects in adult female flies. We found that yolk-GAL4 disrupts adipose tissue integrity and reduces fat body lipid stores, egg production, and resistance to systemic bacterial infections. Knocking down GAL4 expression in yolk-GAL4 heterozygotes using RNAi fully suppressed each of these defects, thus confirming that the GAL4 transgene product induces these phenotypes. Comparing a panel of additional fat body driver lines, we found that GAL4 expression levels directly correlate infection susceptibility, but not with fat levels or egg production. To determine whether other transgenic proteins can impair fat body function, we constructed new fly lines in which the yolk enhancer directs expression of either cytoplasmic or nuclear-localized mCherry, or an alternative transactivator, LexA. We found that only nuclear-localized mCherry and LexA increased infection susceptibility similarly to GAL4, suggesting that intranuclear transgenic proteins in general can curtail the fat bodys induced immune response in a manner highly sensitive to transgene expression strength. Additionally, these new lines can be valuable tools for future studies. More broadly, our findings highlight the potential for transgenes to substantially impact organismal biology and emphasize the importance of rigorously characterizing genetic tools to optimally leverage model systems like Drosophila.

genetics↗

Plumage manipulation alters the integration of social behavior, physiology, internal microbiome, and fitness.

Signals often covary with physiological and behavioral traits to form an axis of integrated phenotypic variation associated with reproductive performance. This pattern of phenotypic integration could result from intrinsic between-individual differences that are causally related to signal production, physiology, and behavior. Alternatively, signal expression itself might generate dynamic feedback between physiology, behavior, and the experienced social environment, resulting in an integrated phenotype. Here, we manipulated the plumage of female tree swallows (Tachycineta bicolor) to decouple the expression of a social signal from any pre-existing behavioral or physiological differences. We collected a time series of physiological samples, monitored social interactions with a sensor network, and tracked reproductive performance. Relative to sham controls, dulled females experienced an altered social environment; overall, these females were visited more by conspecific females and less by males. Dulled females subsequently changed their own behavior by initiating fewer interactions and increasing nestling provisioning. These differences resulted in an altered internal microbiome and glucose levels and, ultimately, dulled females produced more offspring. Moreover, dulled females produced larger clutches than control females in the year after the manipulation. Thus, signal variation alone--independent from any pre-existing differences--had a sustained causal affect on a suite of integrated traits. This finding suggests that dynamic feedback may play an important role in coordinating an integrated signaling phenotype. Our results have implications for understanding how variation in signal expression arises and is maintained and the extent to which the information encoded in signals is contingent upon their use in a social environment.

zoology↗

Insects as phyllosphere microbiome engineers: effects of aphids on a plant pathogen.

Insect herbivores are common in the phyllosphere, the above-ground parts of plants, and encounter diverse plant-associated bacteria there, yet how these organisms interact remains largely unknown. Strains of the bacterium Pseudomonas syringae grow well epiphytically and have been shown to grow within and kill hemipteran insects like the pea aphid, Acyrthosiphon pisum. Aphids are hypothesized to be an alternative host for these epiphytic bacteria but it is unclear if aphids provide fitness benefits to these bacterial pathogens. To determine if epiphytic bacteria could be adapted for infecting aphids, we characterized 21 strains of P. syringae for epiphytic ability and virulence to pea aphids and found that the two traits were positively correlated. For a subset of strains, we tested if the bacteria derived a fitness benefit from the presence of aphids. Some strains benefited significantly, with up to 18.9% higher population densities when aphids were present, and lower starting population density was predictive of higher benefit from aphid presence. However, further investigation found that honeydew, the sugary waste product of aphids, and not growth in aphids, increased P. syringae growth on leaves. This suggests that aphids may be important microbiome engineers in the phyllosphere, but evolutionarily dead-ends for epiphytic bacteria.

evolutionary biology↗