Monoterpenes have an established use in the food and cosmetic industries and have recently also found application as advanced biofuels. Although metabolic engineering efforts have so far achieved significant yields of larger terpenes, monoterpene productivity is lagging behind. Here, we set out to establish a monoterpene-specific production platform in Saccharomyces cerevisiae and identified the sequential reaction mechanism of the yeast farnesyl diphosphate synthase Erg20p to be an important factor limiting monoterpene yield. To overcome this hurdle, we engineered Erg20p into a geranyl diphosphate synthase and achieved a significant increase in monoterpene titers. To further improve production, we converted the engineered geranyl diphosphate synthase into a dominant negative form, so as to decrease the ability of the endogenous Erg20p to function as a farnesyl diphosphate synthase, without entirely abolishing sterol biosynthesis. Fusion of the synthetic dominant negative Erg20p variant with the terpene synthase, combined with yeast strain engineering, further improved monoterpene yields and achieved an overall 340-fold increase in sabinene yield over the starting strain. The design described here can be readily incorporated to any dedicated yeast strain, while the developed plasmid vectors and heterozygous ERG20 deletion yeast strain can also be used as a plug-and-play system for enzyme characterization and monoterpene pathway elucidation.

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The brown marmorated stink bug, Halyomorpha halys (Stål), native to East Asia, emerged as an invasive pest in Europe in the 2000s. In its native range, Trissolcus japonicus (Ashmead) is the dominant egg parasitoid of H. halys, and thus it has been considered for classical biological control in countries invaded by the pest. A survey of native egg parasitoids conducted in 2017 and 2018 with frozen, sentinel egg masses of H. halys revealed that T. japonicus was already present in apple orchards in the Canton Ticino, Switzerland. Trissolcus japonicus was recovered in both years and from three different sites. In total, 17 egg masses were recovered from which 42 adult parasitoids emerged. A genetic analysis using the barcode mitochondrial DNA confirmed the morphological identification of T. japonicus and evidenced a best match of the "Ticino populations" to Japanese populations, but the pathways of entry remain unknown. Keywords Biological control • Egg parasitoids • Halyomorpha halys • Scelionidae Key message • Surveys for egg parasitoids of Halyomorpha halys were conducted in southeastern Switzerland. • For the first time, the Asian parasitoid T. japonicus was recovered from sentinel H. halys egg masses in Europe. • Trissolcus japonicus is established in Switzerland and was found in two consecutive years at three different sites. • Parasitism levels by T. japonicus are currently low, with a maximum of 2%. Communicated by D.C. Weber.

Intracellular reproductive manipulators, such as Candidatus Cardinium and Wolbachia are vertically transmitted to progeny but rarely show co-speciation with the host. In sap-feeding insects, plant tissues have been proposed as alternative horizontal routes of interspecific transmission, but experimental evidence is limited. Here we report results from experiments that show that Cardinium is horizontally transmitted between different phloem sap-feeding insect species through plants. Quantitative PCR and in situ hybridization experiments indicated that the leafhopper Scaphoideus titanus releases Cardinium from its salivary glands during feeding on both artificial media and grapevine leaves. Successional time-course feeding experiments with S. titanus initially fed sugar solutions or small areas of grapevine leaves followed by feeding by the phytoplasma vector Macrosteles quadripunctulatus or the grapevine feeder Empoasca vitis revealed that the symbionts were transmitted to both species. Explaining interspecific horizontal transmission through plants improves our understanding of how symbionts spread, their lifestyle and the symbiont-host intermixed evolutionary pattern.