The physical properties of peat substrates from eight tree nurseries were characterized to determine bulk density, air-filled porosity, saturated hydraulic conductivity, pore effectiveness, relative gas diffusivity and chemical properties. There were significant variations among nurseries both in growth of white spruce [Picea glauca (Moench) Voss] seedlings (1+0) and substrate properties. Shoot dry mass and root collar diameter were negatively correlated with air-filled porosity and saturated hydraulic conductivity, whereas root dry mass was positively correlated with bulk density. Seedling growth increased with increasing substrate bulk density up to ~0.11 g cm−3, above which value conditions may become limiting to plant performance. Our results suggest that there was no growth limitation due to restricted aeration (Ds D0−1 > 0.005 m2 s−1 m−2 s for all substrates except one) and that over-aeration reduced seedling growth under dry irrigation management.

Seedlings are rejected every year in forest nurseries because of insufficient root development and root plug cohesion. Adequate substrate bulk density and aeration properties are of critical importance for root growth and water and nutrient uptake. The quality norms for nursery substrates, based in part on the ratio of coarse to fine particle sizes, have recently been questioned with respect to substrate performance. We conducted experiments in 2008 and 2009 under commercial nursery conditions to determine the effects of substrate physicochemical properties on seedling growth and nutrition. Seedlings were grown in 13 different substrates composed of coarse and fine peat particles (threshold size: 0.5 mm), perlite, and vermiculite, covering a broad range of physical properties with adequate available water and various coarse/fine particle ratios and aeration properties. Substrate properties had significant effects on growth parameters in both years. Seedling growth was affected by water stress in 2008 and low gas diffusivity in 2009. This study emphasizes the importance of management on substrate performance, indicating that short periods of moderate water stress (matric potential lower than −10 kPa) may hinder seedling growth. It also suggests that when irrigated substrates are maintained within the 0 to −5 kPa matric potential range, gas diffusivity should not be lower than 0.003 to 0.005 cm2 s cm−2 s−1, in agreement with the modeled O2 profiles in root plugs. Hence, when designing and using substrates at the nursery scale, more attention should be paid to aeration properties after potting and to the management rather than to the coarse/fine particle ratio.

Spiders that spin orb webs secrete seven types of silk. Although the spinning process of the dragline thread is beginning to be understood, the molecular events that occur in spiders' opisthosomal glands, which produce the other fibers, are unknown due to a lack of data regarding their initial and final structures. Taking advantage of the efficiency of Raman spectromicroscopy in investigating micrometer-sized biological samples, we have determined the secondary structure of proteins in the complete set of glands of the orb-weaving spider Nephila clavipes. The major and minor ampullate silks in the sac of their glands have identical secondary structures typical of natively unfolded proteins. Spidroins are converted into fibers containing highly oriented β-sheets. The capture spiral represents a distinct structural singleton. The proteins are highly disordered prior to spinning and undergo no molecular change or alignment upon spinning. The cylindrical, aciniform, and piriform proteins are folded in their initial state with a predominance of α-helices, but whereas the cylindrical gland forms a fiber similar to the major ampullate thread, the aciniform and piriform glands produce fibers dominated by moderately oriented β-sheets and α-helices. The conformation of the proteins before spinning is related to intrinsic characteristics of their primary structure. Proteins that are unfolded in the gland have repeat sequences composed of submotifs and display no sequence regions with aggregation propensity. By contrast, the folded proteins have neither submotifs nor aggregation-prone sequence regions. Taken together, the Raman data show a remarkable diversity of molecular transformations occurring upon spinning.

Due to its unmatched hardness and chemical inertia, diamond offers many advantages over other materials for extreme conditions and routine analysis by attenuated total reflection (ATR) infrared spectroscopy. Its low refractive index can offer up to a 6-fold absorbance increase compared to germanium. Unfortunately, it also results for strong bands in spectral distortions compared to transmission experiments. The aim of this paper is to present a methodological approach to determine quantitatively the degree of the spectral distortions in ATR spectra. This approach requires the determination of the optical constants (refractive index and extinction coefficient) of the investigated sample. As a typical example, the optical constants of the fibroin protein of the silk worm Bombyx mori have been determined from the polarized ATR spectra obtained using both diamond and germanium internal reflection elements. The positions found for the amide I band by germanium and diamond ATR are respectively 6 and 17 cm(-1) lower than the true value dtermined from the k(nu) spectrum, which is calculated to be 1659 cm(-1). To determine quantitatively the effect of relevant parameters such as the film thickness and the protein concentration, various spectral simulations have also been performed. The use of a thinner film probed by light polarized in the plane of incidence and diluting the protein sample can help in obtaining ATR spectra that are closer to their transmittance counterparts. To extend this study to any system, the ATR distortion amplitude has been evaluated using spectral simulations performed for bands of various intensities and widths. From these simulations, a simple empirical relationship has been found to estimate the band shift from the experimental band height and width that could be of practical use for ATR users. This paper shows that the determination of optical constants provides an efficient way to recover the true spectrum shape and band frequencies of distorted ATR spectra.

Resistance to endoparasitoids in aphids involves complex interactions between insect and microbial players. It is now generally accepted that the facultative bacterial symbiont Hamiltonella defensa of the pea aphid Acyrthosiphon pisum is implicated in its resistance to the parasitoid Aphidius ervi. It has also been shown that heat negatively affects pea aphid resistance, suggesting the thermosensitivity of its defensive symbiosis. Here we examined the effects of heat and UV-B on the resistance of A. pisum to A. ervi and we relate its stability under heat stress to different facultative bacterial symbionts hosted by the aphid. For six A. pisum clones harboring four different facultative symbiont associations, the impact of heat and UV-B was measured on their ability to resist A. ervi parasitism under controlled conditions. The results revealed that temperature strongly affected resistance, while UV-B did not. As previously shown, highly resistant A. pisum clones singly infected with H. defensa became more susceptible to parasitism after exposure to heat. Interestingly, clones that were superinfected with H. defensa in association with a newly discovered facultative symbiont, referred to as PAXS (pea aphid X-type symbiont), not only remained highly resistant under heat stress, but also expressed previously unknown, very precocious resistance to A. ervi compared to clones with H. defensa alone. The prevalence of dual symbiosis involving PAXS and H. defensa in local aphid populations suggests its importance in protecting aphid immunity to parasitoids under abiotic stress.

The relatively high specificity of transgenic plants based on Cry toxins of Bacillus thuringiensis (Bt) implies the possibility of upward agroecosystemic cascades toward new equilibria among arthropods associating with cultivated plants. We examine the hypothesis that exclusion of the Colorado potato beetle from potato expressing the Cry3a toxin increases the abundance of non-target herbivores, which indirectly favours the abundance of herbivore-dependent predators and omnivores foraging on agricultural plants. We examined the impact of Bt potato on non-target arthropod taxa, based on impact studies conducted during development of the Newleaf® Bt potato in North America. Of 32 field tests comparing Bt potato to non-transgenic controls, 14 (42%) revealed a significant, positive effect on the abundance of sucking insects (aphids, leafhoppers, mirids, thrips). Among 72 tests on generalist predators that were simultaneously monitored, 14 (~20%) also revealed significant positive effects. Such positive effects on predators can best be explained by their abundance being increased as a result of greater productivity due to overabundance of sucking insect prey, which are selectively favoured by the high specificity of the Cry3a Bt toxin. Our results support the idea that development of the Bt potato may have been hampered in part by its positive effects on sucking insect pests, and underline the importance of conserving the natural enemies of secondary pests that are indirectly favoured.

The orientational and conformational transformation of the native liquid silk into a solid fiber in the major ampullate gland of the spider Nephila clavipes has been studied by Raman spectromicroscopy. The spectra show that the conformation of silk proteins in the glandular sac contains several secondary structure elements, which is consistent with intrinsically unfolded proteins. A few a-helices are also present and involve some alanine residues located in the polyalanine segments of the spidroin sequence. The conversion of the silk solution in the major ampullate gland appears to be a two-state process without intermediate states. In the first and second limbs of the duct, silk is isotropic and spidroins are generally native-like. beta-Sheets start to develop between the second and the third limb of the duct, suggesting that early beta-sheets are generated by shear forces. However, most of the beta-sheets are formed between the draw down taper and the valve. The early beta-sheets formed upward of the draw down taper might play the role of nucleation sites for the subsequent beta-sheet aggregation. The alignment of the polypeptides chains occurs near the valve, revealing that orientational and conformational changes do not occur simultaneously. Extensional flow seems to be the driving force to produce the orientational order, which in turn is associated with the formation of the major part of the beta-sheets. The slow evolution of the spidroin conformation up to the draw down taper followed by the rapid transformation between the drawn down taper and the valve may be important to achieve the optimal structure of the final fiber.

The relatively high specificity of transgenic plants based on Cry toxins of Bacillus thuringiensis (Bt) implies the possibility of upward agroecosystemic cascades toward new equilibria among arthropods associating with cultivated plants. We examine the hypothesis that exclusion of the Colorado potato beetle from potato expressing the Cry3a toxin increases the abundance of non-target herbivores, which indirectly favours the abundance of herbivore-dependent predators and omnivores foraging on agricultural plants. We examined the impact of Bt potato on non-target arthropod taxa, based on impact studies conducted during development of the Newleaf(R) Bt potato in North America. Of 32 field tests comparing Bt potato to non-transgenic controls, 14 (42%) revealed a significant, positive effect on the abundance of sucking insects (aphids, leathoppers, mirids, thrips). Among 72 tests on generalist predators that were simultaneously monitored, 14 (similar to 20%) also revealed significant positive effects. Such positive effects on predators can best be explained by their abundance being increased as a result of greater productivity due to overabundance of sucking insect prey, which are selectively favoured by the high specificity of the Cry3a Bt toxin. Our results support the idea that development of the Bt potato may have been hampered in part by its positive effects on sucking insect pests, and underline the importance of conserving the natural enemies of secondary pests that are indirectly favoured.

Host insects are either susceptible or resistant to parasitoids, where resistant hosts express immunity factors and compatible parasitoids express virulence factors that may reveal the manipulation of susceptible hosts. Using proteomics we compared responses of the same host, the aphid Macrosiphum euphorbiae, challenged by a well-adapted parasitoid Aphidius nigripes or by a less adapted relative, Aphidius ervi. The host was found to be equally acceptable to both parasitoids, but while A. nigripes normally developed and killed hosts (high susceptibility), development of the incompatible A. ervi was arrested at the primary egg stage (high resistance). Two-dimensional gels at two stages of parasitism revealed divergence in patterns of protein regulation of the M. euphorbiae host, responding to A. ervi or A. nigripes, with the greatest number of protein modulations in the host resistance response. In A. ervi-resistant hosts, proPO was strongly up-regulated, as were also three cuticle proteins, suggesting a PO basis and exoskeleton reinforcement as early and late responses of M. euphorbiae to the risk of parasitism. Resistance also correlated with up-regulation of antioxidative, energy-related, cytoskeleton and heat shock proteins. In A. nigripes-susceptible hosts, various proteins implicated in host and bacterial symbiont metabolism were significantly altered, suggesting complex host nutritional modulation. Over-expression of energy-related proteins also increased when A. nigripes established and developed. Aphid proteomes of compatible and incompatible Aphidius parasitism provide an integrative basis for consolidating our knowledge of host–parasitoid interactions.

To understand the spinning process of dragline silk by spiders, the protein conformation before spinning has to be determined. Raman confocal spectromicroscopy has been used to study the conformation of the proteins in situ in the intact abdominal major ampullate gland of Nephila clavipes and Araneus diadernatus spiders. The spectra obtained are typical of natively unfolded proteins and are very similar to that of a mixture of recombinant silk proteins. Vibrational circular dichroism reveals that the conformation is composed of random and polyproline II (PPII) segments with some a-helices. The a-helices seem to be located in the C-terminal part whereas the repetitive sequence is unfolded. The PPII structure can significantly contribute to the efficiency of the spinning process in nature.

Modeling the effect of temperature on the sustainability of insect–plant interactions requires assessment of both insect and plant performance. We examined the effect of temperature on western flower thrips, Frankliniella occidentalis (Pergande), a generalist herbivore with a high reproductive rate, and chrysanthemum inflorescences, a high quality but relatively fixed, ephemeral resource for thrips population growth. We hypothesized that different thrips versus plant responses to temperature result in significant statistical interaction of temperature with thrips abundance and flower damage attributes over time. Experiments were conducted at five temperatures between 20.7 and 35.3°C, with thrips infestation and time after infestation as main effects. Only minor, uncontrolled variations in relative humidity and light intensity may otherwise have influenced the results. High temperatures lead to an initially rapid increase in density of thrips followed by abrupt declines in abundance. The rate of floral senescence increased with temperature and thrips infestation, as indicated by a reduced fresh biomass and greater leaching of yellow pigments. Multiple regression indicated that indices of plant damage responded more directly to thrips density at low than high temperature, supporting the conclusion that temperature affected the outcome beyond what was predictable simply from differential plant and insect optima. The relative intensity of damage caused by individual thrips decreased with increasing temperature, likely caused by thrips competition and reduced survival, growth, and fecundity on depleted inflorescences. Reduced per capita damage at high temperature may be common in insects exploiting fixed plant resources that exhibit an accelerated rate of deterioration at high temperatures.

Parasitoid virulence and host resistance are complex interactions depending on metabolic rate and cellular activity, which in aphids additionally implicate heritable secondary symbionts among the Enterobacteriaceae. As performance of the parasitoid, the aphid host and its symbionts may differentially respond to temperature, the success or failure of aphid parasitism is difficult to predict when temperature varies. We tested the hypothesis that resistance of the pea aphid Acyrthosiphon pisum to the parasitoid Aphidius ervi, which is linked to aphid secondary symbionts, may depend on temperature in several resistant and non-resistant aphid clonal lineages of different geographic origin and of known bacterial symbiosis, using experiments in controlled environments. Complete immunity to A. ervi at 20°C in three different aphid clones whose symbiosis is characterized by the possession of Hamiltonella defensa reversed to high susceptibility at 25°C and especially 30°C, suggesting that the aphid's immune responses to the establishment and early development of the parasitoid is strongly reduced at moderately high temperatures. There was no evidence that a pea aphid control genotype that was susceptible to A. ervi at 20°C could become more resistant as temperature increases, as has been suggested for insect fungal pathogens. By contrast, our results suggest that aphid clonal resistance to A. ervi and related parasitoids is characteristic of cool temperature conditions, similar to various other fitness attributes of aphids. Based on evidence that H. defensa symbionts characterized all three A. ervi resistant pea aphid clones studied, but was absent in control aphids that remained susceptible at all temperatures, we suggest that secondary symbiosis plays a key role in the heat sensitivity of aphid clonal resistance. Our study may also indicate that aphid natural control of variably susceptible host populations by aphid parasitoids is more likely at moderate to high temperatures.