RESUMO
Cereal-feeding beetles are a major risk for cereal crop maintenance. Cereal weevils such as Sitophilus oryzae have symbiotic intracellular bacteria that provide essential aromatic amino acid to the host for the biosynthesis of their cuticle building blocks. Their cuticle is an important protective barrier against biotic and abiotic stresses, providing high resistance from insecticides. Quantitative optical methods specialized in insect cuticle analysis exist, but their scope of use and the repeatability of the results remain limited. Here, we investigated the potential of Hyperspectral Imaging (HSI) as a standardized cuticle analysis tool. Based on HSI, we acquired time series of average reflectance profiles from 400 to 1000 nm from symbiotic (with bacteria) and aposymbiotic (without bacteria) cereal weevils S. oryzae exposed to different nutritional stresses. We assessed the phenotypic changes of weevils under different diets throughout their development and demonstrated the agreement of the results between the HSI method and the classically used Red-Green-Blue analysis. Then, we compared the use of both technologies in laboratory conditions and highlighted the assets of HSI to develop a simple, automated, and standardized analysis tool. This is the first study showing the reliability and feasibility of HSI for a standardized analysis of insect cuticle changes.
RESUMO
Nutritional symbioses between insects and intracellular bacteria (endosymbionts) are a major force of adaptation, allowing animals to colonize nutrient-poor ecological niches. Many beetles feeding on tyrosine-poor substrates rely on a surplus of aromatic amino acids produced by bacterial endosymbionts. This surplus of aromatic amino acids is crucial for the biosynthesis of a thick exoskeleton, the cuticle, which is made of a matrix of chitin with proteins and pigments built from tyrosine-derived molecules, providing an important defensive barrier against biotic and abiotic stress. Other endosymbiont-related advantages for beetles include faster development and improved fecundity. The association between Sitophilus oryzae and the Sodalis pierantonius endosymbiont represents a unique case study among beetles: endosymbionts undergo an exponential proliferation in young adults concomitant with the cuticle tanning, and then they are fully eliminated. While endosymbiont clearance, as well as total endosymbiont titer, are host-controlled processes, the mechanism triggering endosymbiont exponential proliferation remains poorly understood. Here, we show that endosymbiont exponential proliferation relies on host carbohydrate intake, unlike the total endosymbiont titer or the endosymbiont clearance, which are under host genetic control. Remarkably, insect fecundity was preserved, and the cuticle tanning was achieved, even when endosymbiont exponential proliferation was experimentally blocked, except in the context of a severely unbalanced diet. Moreover, a high endosymbiont titer coupled with nutrient shortage dramatically impacted host survival, revealing possible environment-dependent disadvantages for the host, likely due to the high energy cost of exponentially proliferating endosymbionts. IMPORTANCE Beetles thriving on tyrosine-poor diet sources often develop mutualistic associations with endosymbionts able to synthesize aromatic amino acids. This surplus of aromatic amino acids is used to reinforce the insect's protective cuticle. An exceptional feature of the Sitophilus oryzae/Sodalis pierantonius interaction is the exponential increase in endosymbiotic titer observed in young adult insects, in concomitance with cuticle biosynthesis. Here, we show that host carbohydrate intake triggers endosymbiont exponential proliferation, even in conditions that lead to the detriment of the host survival. In addition, when hosts thrive on a balanced diet, endosymbiont proliferation is dispensable for several host fitness traits. The endosymbiont exponential proliferation is therefore dependent on the nutritional status of the host, and its consequences on host cuticle biosynthesis and survival depend on food quality and availability.
