Inka Lusebrink

Diesel exhaust rapidly degrades floral odours used by honeybees

Honeybees utilise floral odours when foraging for flowers; we investigated whether diesel exhaust pollution could interrupt these floral odour stimuli. A synthetic blend of eight floral chemicals, identified from oilseed rape, was exposed to diesel exhaust pollution. Within one minute of exposure the abundances of four of the chemicals were significantly lowered, with two components rendered undetectable. Honeybees were trained to recognise the full synthetic odour mix; altering the blend, by removing the two chemicals rendered undetectable, significantly reduced the ability of the trained honeybees to recognize the altered odour. Furthermore, we found that at environmentally relevant levels the mono-nitrogen oxide (NOx) fraction of the exhaust gases was a key facilitator of this odour degradation. Such changes in recognition may impact upon a honeybees foraging efficiency and therefore the pollination services that they provide.

Variation in carbon availability, defense chemistry and susceptibility to fungal invasion along the stems of mature trees

If carbon (C) sinks withdraw carbohydrates as they are transported along tree stems, carbohydrate availability may depend on local sink strength and distance from sources. Defenses, including monoterpenes--a major component of resin--limit the invasibility of pines. Since carbohydrate reserves fund monoterpene synthesis, we hypothesized that monoterpene concentrations in pine stems would decrease from the crown to the lower stem, and susceptibility to fungal infection would increase. Here, we measured carbohydrate and monoterpene concentrations along the stems of lodgepole pine trees (Pinus contorta var. latifolia) before inoculating with a blue-stain fungus at different heights. After 6 wk, we assessed tree responses to fungal infection based on lesion length and carbohydrate mobilization. Concentrations of carbohydrates and monoterpenes in the phloem before inoculation decreased with distance from the crown, whereas lesion lengths after inoculation increased. However, trees mobilized sugars in response to fungal infection such that carbohydrate reserves near lesions were similar at all heights. Despite C mobilization, the lower stem was more vulnerable than the upper stem. Consistent with predictions based on sink-source relationships, vulnerability occurred where carbohydrates were less available, and likely resulted from C withdrawal by sinks higher in the supply chain.

Stenusine, an antimicrobial agent in the rove beetle genus Stenus (Coleoptera, Staphylinidae)

Stenusine is well known as the alkaloid, discharged by the rove beetle, genus Stenus Latreille (Coleoptera, Staphylinidae). The Stenus beetles employ the alkaloid as an escape mechanism when on water surfaces. In the case of danger, they lower their abdomen and emit stenusine from their pygidial glands. Stenusine shows a low surface tension and therefore a high spreading pressure; these properties propel the beetle quickly over the water. Many Steninae do not live in habitats with open waters, but in detritus, leaf litter, mosses, etc. This raises the possibility that stenusine might also have another function, e.g., as antibiotic or fungicide. Stenus beetles show an intense grooming behaviour. With gas chromatography–mass spectrometry analyses we could prove that they cover themselves with their secretion. To tests its antimicrobial properties we conducted agar diffusion tests with stenusine and norstenusine, another substance that is abundant in most Stenus species. Both compounds have an antimicrobial effect on entomopathogenic bacteria and fungi. Stenusine not only allows for an extraordinary method of locomotion on water surfaces, it also protects the Steninae from being infested with microorganisms.

Differences in defence responses of Pinus contorta and Pinus banksiana to the mountain pine beetle fungal associate Grosmannia clavigera are affected by water deficit

We tested the hypotheses that responses to the mountain pine beetle fungal associate Grosmannia clavigera will differ between the evolutionarily co-evolved host lodgepole pine (Pinus contorta var. latifolia) and the naïve host jack pine (Pinus banksiana) and that these responses will be influenced by water availability. G. clavigera inoculation resulted in more rapid stem lesion development in lodgepole than in jack pine; water deficit delayed lesion development in both species. Decreased hydraulic conductivity was observed in inoculated lodgepole pine seedlings, likely because of tracheid occlusion by fungal hyphae and/or metabolite accumulation. Drought but not inoculation significantly impacted bark abscisic acid levels. Jasmonic and salicylic acid were implicated in local and systemic responses of both species to G. clavigera, with salicylic acid appearing to play a greater role in jack pine response to G. clavigera than lodgepole pine. Water deficit increased constitutive levels and/or attenuated induced responses to G. clavigera for several monoterpenes in lodgepole but not jack pine. Instead, inoculation of well-watered but not water deficit jack pine resulted in a greater number of xylem resin ducts. These findings reveal mechanisms underlying differences in G. clavigera-induced responses between lodgepole and jack pine hosts, and how water availability modulates these responses.

The lodgepole × jack pine hybrid zone in Alberta, Canada: a stepping stone for the mountain pine beetle on its journey East across the boreal forest?

Historical data show that outbreaks of the tree killing mountain pine beetle are often preceded by periods of drought. Global climate change impacts drought frequency and severity and is implicated in the range expansion of the mountain pine beetle into formerly unsuitable habitats. Its expanded range has recently reached the lodgepole × jack pine hybrid zone in central Alberta, Canada, which could act as a transition from its historical lodgepole pine host to a jack pine host present in the boreal forest. This field study tested the effects of water limitation on chemical defenses of mature trees against mountain pine beetle-associated microorganisms and on beetle brood success in lodgepole × jack pine hybrid trees. Tree chemical defenses as measured by monoterpene emission from tree boles and monoterpene concentration in needles were greater in trees that experienced water deficit compared to well-watered trees. Myrcene was identified as specific defensive compound, since it significantly increased upon inoculation with dead mountain pine beetles. Beetles reared in bolts from trees that experienced water deficit emerged with a higher fat content, demonstrating for the first time experimentally that drought conditions benefit mountain pine beetles. Further, our study demonstrated that volatile chemical emission from tree boles and phloem chemistry place the hybrid tree chemotype in-between lodgepole pine and jack pine, which might facilitate the host shift from lodgepole pine to jack pine.

The Effect of Water Limitation on Volatile Emission, Tree Defense Response, and Brood Success of Dendroctonus ponderosae in Two Pine Hosts, Lodgepole, and Jack Pine

The mountain pine beetle (MPB; Dendroctonus ponderosae) has recently expanded its range from lodgepole pine forest into the lodgepole × jack pine hybrid zone in central Alberta, within which it has attacked pure jack pine. This study tested the effects of water limitation on tree defense response of mature lodgepole and jack pine (Pinus contorta and Pinus banksiana) trees in the field. Tree defense response was initiated by inoculation of trees with the MPB-associated fungus Grosmannia clavigera and measured through monoterpene emission from tree boles and concentration of defensive compounds in phloem, needles, and necrotic tissues. Lodgepole pine generally emitted higher amounts of monoterpenes than jack pine; particularly from fungal-inoculated trees. Compared to non-inoculated trees, fungal inoculation increased monoterpene emission in both species, whereas water treatment had no effect on monoterpene emission. The phloem of both pine species contains (-)-α-pinene, the precursor of the beetle’s aggregation pheromone, however lodgepole pine contains two times as much as jack pine. The concentration of defensive compounds was 70-fold greater in the lesion tissue in jack pine, but only 10-fold in lodgepole pine compared to healthy phloem tissue in each species, respectively. Water-deficit treatment inhibited an increase of L-limonene as response to fungal inoculation in lodgepole pine phloem. The amount of myrcene in jack pine phloem was higher in water-deficit trees compared to ambient trees. Beetles reared in jack pine were not affected by either water or biological treatment, whereas beetles reared in lodgepole pine benefited from fungal inoculation by producing larger and heavier female offspring. Female beetles that emerged from jack pine bolts contained more fat than those that emerged from lodgepole pine, even though lodgepole pine phloem had a higher nitrogen content than jack pine phloem. These results suggest that jack pine chemistry is suitable for MPB pheromone production and aggregation on the host tree.

The Effects of Diesel Exhaust Pollution on Floral Volatiles and the Consequences for Honey Bee Olfaction

There is growing evidence of a substantial decline in pollinators within Europe and North America, most likely caused by multiple factors such as diseases, poor nutrition, habitat loss, insecticides, and environmental pollution. Diesel exhaust could be a contributing factor to this decline, since we found that diesel exhaust rapidly degrades floral volatiles, which honey bees require for flower recognition. In this study, we exposed eight of the most common floral volatiles to diesel exhaust in order to investigate whether it can affect volatile mediated plant-pollinator interaction. Exposure to diesel exhaust altered the blend of common flower volatiles significantly: myrcene was considerably reduced, β-ocimene became undetectable, and β-caryophyllene was transformed into its cis-isomer isocaryophyllene. Proboscis extension response (PER) assays showed that the alterations of the blend reduced the ability of honey bees to recognize it. The chemically reactive nitrogen oxides fraction of diesel exhaust gas was identified as capable of causing degradation of floral volatiles.

Influence of water deficit on the molecular responses of Pinus contorta × Pinus banksiana mature trees to infection by the mountain pine beetle fungal associate, Grosmannia clavigera.

Conifers exhibit a number of constitutive and induced mechanisms to defend against attack by pests and pathogens such as mountain pine beetle (Dendroctonus ponderosae Hopkins) and their fungal associates. Ecological studies have demonstrated that stressed trees are more susceptible to attack by mountain pine beetle than their healthy counterparts. In this study, we tested the hypothesis that water deficit affects constitutive and induced responses of mature lodgepole pine × jack pine hybrids (Pinus contorta Dougl. ex Loud. var. latifolia Engelm. ex S. Wats. × Pinus banksiana Lamb.) to inoculation with the mountain pine beetle fungal associate Grosmannia clavigera (Robinson-Jeffrey and Davidson) Zipfel, de Beer and Wingfield. The degree of stress induced by the imposed water-deficit treatment was sufficient to reduce photosynthesis. Grosmannia clavigera-induced lesions exhibited significantly reduced dimensions in water-deficit trees relative to well-watered trees at 5 weeks after inoculation. Treatment-associated cellular-level changes in secondary phloem were also observed. Quantitative RT-PCR was used to analyze transcript abundance profiles of 18 genes belonging to four families classically associated with biotic and abiotic stress responses: aquaporins (AQPs), dehydration-responsive element binding (DREB), terpene synthases (TPSs) and chitinases (CHIs). Transcript abundance profiles of a TIP2 AQP and a TINY-like DREB decreased significantly in fungus-inoculated trees, but not in response to water deficit. One TPS, Pcb(+)-3-carene synthase, and the Class II CHIs PcbCHI2.1 and PcbCHI2.2 showed increased expression under water-deficit conditions in the absence of fungal inoculation, while another TPS, Pcb(E)-β-farnesene synthase-like, and two CHIs, PcbCHI1.1 and PcbCHI4.1, showed attenuated expression under water-deficit conditions in the presence of fungal inoculation. The effects were observed both locally and systemically. These results demonstrate that both constitutive and induced carbon- and nitrogen-based defenses are affected by water deficit, suggesting potential consequences for mountain pine beetle dynamics, particularly in novel environments.

New pyridine alkaloids from rove beetles of the genus Stenus (Coleoptera: Staphylinidae).

Three new pyridine alkaloids were detected in the pygidial glands of some Stenus species. The chemotaxonomic significance of the occurrence of these alkaloids and stenusine in different Stenus species is discussed. The antimicrobial properties of (Z)- and (E)-3-(2- methyl-1-butenyl)-pyridine and the deterrent activities of stenusine and norstenusine were investigated.

Biosynthesis of Stenusine

The rove beetles of the genus Stenus Latreille synthesize the alkaloid stenusine in their pygidial glands, which are located in the last three segments of their abdomen. It is proposed that stenusine is derived from the two amino acids, L-lysine and L-isoleucine. Feeding S. bimaculatus beetles with deuterium-labeled amino acids and using GC/MS analysis showed that L-lysine forms the piperidine ring of the stenusine molecule. The side chain originates from L-isoleucine and the N-ethyl group from acetate.