Field assessments and genetic analyses on tansy aphid, Metopeurum fuscoviride Stroyan were made aiming to investigate whether a specialist aphid species and its associated predator density differ between chemotypes of the plant host, tansy, and whether the essential oil composition of the plant host determines the genetic differences observed between colonies of M. fuscoviride inhabiting these different chemotypes. Tansy plants were defined after volatile extractions as Camphor Pure and Borneol Pure, respectively, with hybrid chemotypes defined as Camphor Hybrid, Borneol Hybrid, and Thujone Hybrid, respectively. Ladybird beetles, Coccinella septempunctata (L.), clearly dominated Pure Camphor plants, whereas significantly higher numbers of the nursery web spider, Pisaura mirabilis (Clerck), were detected on Borneol Pure and Borneol Hybrid plants. Genetic analyses using five polymorphic microsatellite markers revealed that higher genetic similarity existed between M. fuscoviride aphids from Borneol Pure and Borneol Hybrid plants and between Camphor Pure and Camphor Hybrid plants, respectively, whereas a lower level of genetic similarity was seen between these aphids from Thujone Hybrid plants. Accordingly, it is possible that host plant‐adapted species can create much context‐dependency in the nature and strength of predation effects, which may in turn alter costs and benefits of host plant chemical differentiations, or even the prey genotype variations. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 114, 709–719.

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Two histidines were introduced by site-directed mutagenesis into the structure of Enhanced Green Fluorescent Protein, replacing the serine at position 202 and the glutamine at position 204 for increasing the sensitivity of the protein towards different metal ions by creating possible metal binding sites near the chromophore group. There is no appreciable difference between the absorbance and fluorescence spectra of the two proteins (wild type and the double-histidine mutant) indicating that the mutation does not change the environment of the fluorophore. Fluorescence quenching was measured at different pH (6.5-8) and temperatures (20-45 °C) varying the concentration of metal ions. Under optimal conditions (pH = 7.5, 20 °C) the mutant's Kd is 16 nM, it binds copper more than 200fold stronger than the wild type EGFP.

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The histidine-modified EGFP was characterized as a sensing element that preferentially binds nanomolar concentrations of Cu(2+) in a reversible manner (Kd = 15 nM). This research aims to determine the causes of nanomolar-affinity of this mutant by investigating significant structural and energetic alterations of the chromophore in the presence of different copper ion concentrations. In order to reveal the unknown parts of the quenching mechanism we have elaborated a specific approach that combines theoretical and experimental techniques. The theoretical experiment included the modeling of potential distortions of the chromophores and the corresponding changes in energy using quantum mechanical calculations. Differences between the modeled energy profiles of planar and distorted conformations represented the energies of activation for the chromophore distortions. We found that some values of the experimental activation energies, which were derived from fluorescence lifetime decay analysis (ex: 470 nm, em: 507 nm), were consistent with the theoretical ones. Thus, it has been revealed similarity between the theoretical activation energy (50 kJmol(-1)) for 40° phenolate-ring distortion and the experimental activation energy (52.17 kJmol(-1)) required for histidine-modified EGFP saturation with copper. This chromophore conformation was further investigated and it has been found that the large decrease in fluorescence emission is attributed to the significant charge transfer over the molecule which triggers proton transfer thereby neutralizing the cromophore.