A blood test to monitor bee health across a European network of agricultural sites of different land-use by MALDI BeeTyping mass spectrometry.

There are substantial concerns about impaired honey bee health and colony losses due to several poorly understood factors. We used MALDI profiling (MALDI BeeTyping®) analysis to investigate how some environmental and management factors under field conditions across Europe affected the honey bee haemolymph peptidome (all peptides in the circulatory fluid), as a profile of molecular markers representing the immune status of Apis mellifera. Honey bees were exposed to a range of environmental stressors in 128 agricultural sites across eight European countries in four biogeographic zones, with each country contributing eight sites each for two different cropping systems: oilseed rape (OSR) and apple (APP). The full haemolymph peptide profiles, including the presence and levels of three key immunity markers, namely the antimicrobial peptides (AMPs) Apidaecin, Abaecin and Defensin-1, allowed the honey bee responses to environmental variables to be discriminated by country, crop type and site. When considering just the AMPs, it was not possible to distinguish between countries by the prevalence of each AMP in the samples. However, it was possible to discriminate between countries on the amounts of the AMPs, with the Swedish samples in particular expressing high amounts of all AMPs. A machine learning model was developed to discriminate the haemolymphs of bees from APP and OSR sites. The model was 90.6 % accurate in identifying the crop type from the samples used to build the model. Overall, MALDI BeeTyping® of bee haemolymph represents a promising and cost-effective "blood test" for simultaneously monitoring dozens of peptide markers affected by environmental stressors at the landscape scale, thus providing policymakers with new diagnostic and regulatory tools for monitoring bee health.

Response of Pasture Grasses to Organic Fertilizer Produced from Black Soldier Fly Frass.

Livestock and dairy farmers are increasingly required to maintain productivity and profitability while mitigating the environmental harm associated with high-input agriculture. Accordingly, to reduce reliance on synthetic fertilizers, a wide range of organically derived products are being evaluated for their effects on sward growth and forage quality. This study used glasshouse experiments to investigate the responses of four grass species to a novel organic fertilizer derived from the mass production of black soldier fly larvae [Hermetia illucens, HexaFrass™, Meath, Ireland]. Although there was some variability among trials, overall our results indicate that application of HexaFrass produced increased shoot growth of Perennial Ryegrass (PRG; Lolium perenne L.), Timothy (Phleum pratense L.), and Cocksfoot (Dactylis glomerata L.) compared with growth achieved in no-fertilizer control plants. In addition to increases in shoot fresh and dry weight, shoot chlorophyll content was also positively related to the HexaFrass application rate. At nitrogen-equivalent application rates, HexaFrass resulted in lower shoot growth compared with the application of urea, suggesting that the nitrogen contained in HexaFrass may not be immediately plant-available. Of relevance to grazing or silage systems, the addition of HexaFrass resulted in increased shoot regrowth of PRG and Timothy after shoots had been cut. Based on our results, insect-frass-based fertilizers may have a role in low input, organic, and/or regenerative pasture-based livestock systems, although issues may occur due to the relatively high costs and low availability compared with other organic soil amendments such as farmyard manure and slurry.

Novel indices reveal that pollinator exposure to pesticides varies across biological compartments and crop surroundings.

Declines in insect pollinators have been linked to a range of causative factors such as disease, loss of habitats, the quality and availability of food, and exposure to pesticides. Here, we analysed an extensive dataset generated from pesticide screening of foraging insects, pollen-nectar stores/beebread, pollen and ingested nectar across three species of bees collected at 128 European sites set in two types of crop. In this paper, we aimed to (i) derive a new index to summarise key aspects of complex pesticide exposure data and (ii) understand the links between pesticide exposures depicted by the different matrices, bee species and apple orchards versus oilseed rape crops. We found that summary indices were highly correlated with the number of pesticides detected in the related matrix but not with which pesticides were present. Matrices collected from apple orchards generally contained a higher number of pesticides (7.6 pesticides per site) than matrices from sites collected from oilseed rape crops (3.5 pesticides), with fungicides being highly represented in apple crops. A greater number of pesticides were found in pollen-nectar stores/beebread and pollen matrices compared with nectar and bee body matrices. Our results show that for a complete assessment of pollinator pesticide exposure, it is necessary to consider several different exposure routes and multiple species of bees across different agricultural systems.

Distribution of infectious and parasitic agents among three sentinel bee species across European agricultural landscapes.

Infectious and parasitic agents (IPAs) and their associated diseases are major environmental stressors that jeopardize bee health, both alone and in interaction with other stressors. Their impact on pollinator communities can be assessed by studying multiple sentinel bee species. Here, we analysed the field exposure of three sentinel managed bee species (Apis mellifera, Bombus terrestris and Osmia bicornis) to 11 IPAs (six RNA viruses, two bacteria, three microsporidia). The sentinel bees were deployed at 128 sites in eight European countries adjacent to either oilseed rape fields or apple orchards during crop bloom. Adult bees of each species were sampled before their placement and after crop bloom. The IPAs were detected and quantified using a harmonised, high-throughput and semi-automatized qPCR workflow. We describe differences among bee species in IPA profiles (richness, diversity, detection frequencies, loads and their change upon field exposure, and exposure risk), with no clear patterns related to the country or focal crop. Our results suggest that the most frequent IPAs in adult bees are more appropriate for assessing the bees' IPA exposure risk. We also report positive correlations of IPA loads supporting the potential IPA transmission among sentinels, suggesting careful consideration should be taken when introducing managed pollinators in ecologically sensitive environments.

Pesticide use negatively affects bumble bees across European landscapes.

Sustainable agriculture requires balancing crop yields with the effects of pesticides on non-target organisms, such as bees and other crop pollinators. Field studies demonstrated that agricultural use of neonicotinoid insecticides can negatively affect wild bee species1,2, leading to restrictions on these compounds3. However, besides neonicotinoids, field-based evidence of the effects of landscape pesticide exposure on wild bees is lacking. Bees encounter many pesticides in agricultural landscapes4-9 and the effects of this landscape exposure on colony growth and development of any bee species remains unknown. Here we show that the many pesticides found in bumble bee-collected pollen are associated with reduced colony performance during crop bloom, especially in simplified landscapes with intensive agricultural practices. Our results from 316 Bombus terrestris colonies at 106 agricultural sites across eight European countries confirm that the regulatory system fails to sufficiently prevent pesticide-related impacts on non-target organisms, even for a eusocial pollinator species in which colony size may buffer against such impacts10,11. These findings support the need for postapproval monitoring of both pesticide exposure and effects to confirm that the regulatory process is sufficiently protective in limiting the collateral environmental damage of agricultural pesticide use.

The Effects of Two Organic Soil Amendments, Biochar and Insect Frass Fertilizer, on Shoot Growth of Cereal Seedlings.

To mitigate the environmental harm associated with high-input agriculture, arable farmers are increasingly required to maintain productivity while reducing inputs of synthetic fertilizers. Thus, a diverse range of organic products are now being investigated in terms of their value as alternative fertilizers and soil amendments. This study used a series of glasshouse trials to investigate the effects of an insect frass-based fertilizer derived from black soldier fly waste [HexaFrass™, Meath, Ireland] and biochar on four cereals grown in Ireland (barley, oats, triticale, spelt) as animal feed and for human consumption. In general, the application of low quantities of HexaFrass™ resulted in significant increases in shoot growth in all four cereal species, along with increased foliage concentrations of NPK and SPAD levels (a measure of chlorophyll density). These positive effects of HexaFrass™ on shoot growth were observed, however, only when a potting mix with low basal nutrients was used. Additionally, excessive application of HexaFrass™ resulted in reduced shoot growth and, in some cases, seedling mortality. The application of finely ground or crushed biochar produced from four different feedstocks (Ulex, Juncus, woodchip, olive stone) had no consistent positive or negative effects on cereal shoot growth. Overall, our results indicate that insect frass-based fertilizers have good potential in low-input, organic, or regenerative cereal production systems. Based on our results, biochar appears to have less potential as a plant growth promoting product, but could be used as a tool for lowering whole-farm carbon budgets by providing a simplistic means of storing carbon in farm soils.

Responses in honeybee and bumblebee activity to changes in weather conditions.

Insect pollination, and in particular pollination by bees, is a highly valued ecosystem service that ensures plant reproduction and the production of high-quality crops. Bee activity is known to be influenced by the weather, and as the global climate continues to change, the flying frequency and foraging behaviour of bees may also change. To maximise the benefits of pollination in a changing world, we must first understand how current weather conditions influence the activity of different bee species. This is of particular interest in a country such as Ireland where inclement weather conditions are nominally sub-optimal for foraging. We observed honeybee (Apis mellifera) and buff-tailed bumblebee (Bombus terrestris) activity across a variety of weather conditions at seven apple orchards to determine how four weather variables (temperature, relative humidity, solar radiation, wind) influenced the flight activity of each species. Each orchard contained three honeybee and three bumblebee colonies, and so we were able to observe a colony of each species concurrently in the same weather conditions. Overall, honeybees were more sensitive to changes in weather than bumblebees and could be more predisposed to future changes in within-day weather conditions. Our results indicate bumblebees could compensate for low honeybee activity in inclement conditions, which supports the theory that pollinator diversity provides resilience. This may be particularly important in management of pollinators in crops that flower in the spring when weather is more variable, and to allow varied responses to global climate change.

Glasshouse Evaluation of the Black Soldier Fly Waste Product HexaFrass™ as an Organic Fertilizer.

The mass farming of the black soldier fly (BSF; Hermetia illucens L.), to produce insect-based feed for livestock and fish, results in considerable amounts of insect frass, which contains substantial amounts of organic matter and bioavailable nutrients. Insect frass has shown good potential as a soil amendment and organic fertilizer. This study examined the effects of HexaFrass™ on the growth of common vegetables and herbs under glasshouse conditions. In an organically-rich potting mix, HexaFrass™ increased shoot dry weight by an average of 25%, although this effect was variable among test plants. In other trials, application of HexaFrass™ caused an increase in plant growth similar to that obtained by applying chicken manure and a commercial organic fertilizer. Increases in shoot and root dry weight showed quadratic relationships with HexaFrass™ dose, indicating that application of excessive quantities could lead to plant inhibition. Shoot:root dry matter ratio tended to increase with HexaFrass™ dose suggesting there was no specific stimulation or enhancement of root growth. Overall, these results provide further evidence of the potential of insect frass as an effective organic fertilizer for vegetables and herbs.

Seedling Responses to Organically-Derived Plant Growth Promoters: An Effects-Based Approach.

Organically-derived biofertilizers and biostimulants, developed from harvested materials such as seaweed and waste from animal and fish processing, are currently the subject of much fundamental and applied research. These products have significant potential in reducing synthetic fertilizer inputs to horticultural, arable, and pasture-based agricultural systems, although there is frequently some ambiguity over the magnitude and consistency of any positive effects these products may have on plant performance. This study examined the effects of organically-derived plant growth promoters (PGPs) available in New Zealand on the early vegetative growth of 16 plant species maintained under glasshouse conditions. When applied as a root drench to low nutrient potting mix, the effects of the PGPs on seedling shoot growth were strongly related to the NPK contents of the applied solutions. Any positive effects on shoot growth were, on average, reduced when the seedlings were maintained in higher nutrient growing media. Applying the PGPs at concentrations twice, and four times, the recommended concentration, only caused further growth responses when the PGPs contained high levels of nutrients. Applying the PGPs as a foliar spray had negligible effects on shoot growth. Overall, the results of these trials suggest that the positive effects of applying some organically-derived PGPs on seedling growth are a function of the PGP nutrient content, and not due to any indirect effects related to phytohormone pathways or modification of rhizosphere microorganisms.

Integrating Plantain (Plantago lanceolata L.) and Italian Ryegrass (Lolium multiflorum Lam.) into New Zealand Grazing Dairy System: The Effect on Farm Productivity, Profitability, and Nitrogen Losses.

A two-year farm system study was conducted at Canterbury, New Zealand to evaluate the effects on farm productivity, profitability, and nitrogen (N) losses of integrating plantain (Plantago lanceolate L.) and Italian ryegrass (Lolium multiflorum Lam.) into a ryegrass and white clover (RGWC)-based dairy system. Three farm systems were compared: (1) a lower input RGWC-based system (LIRG) with stocking rate of 3.5 cow/ha, annual N fertiliser rate of 150 kg/ha, and imported feed level of <1.2 t DM/cow/year; (2) a lower input ryegrass + plantain-based system (LIRG + PL) with a stocking rate of 3.5 cow/ha, annual N fertiliser rate of 150 kg/ha, and imported feed level of <1.2 t DM/cow/year; and (3) a higher input RGWC-based system (HIRG) with a stocking rate of 5.0 cow/ha, annual N fertiliser rate of 300 kg/ha, and imported feed level of >1.2 t DM/cow/year. Cows in the LIRG + PL system grazed a diverse mix of Italian ryegrass, perennial ryegrass, white clover, and plantain (60% of farmlet area), and a mixed sward of plantain-white clover (40% of farmlet area). The average annual herbage harvested was similar between LIRG + PL and LIRG (11.7 t DM/ha), but greater in HIRG (12.7 t DM/ha) with the increased N fertiliser rate. During the calving to dry-off period, the average imported supplement feed per ha was higher in HIRG (8.0 t DM) compared with LIRG (3.2 t DM) and LIRG + PL (3.7 t DM). Average milk solid production (MS; fat + protein) was similar in LIRG + PL (1640 kg/ha) and LIRG (1622 kg/ha), but greater in HIRG (2130 kg/ha). Estimated profitability (NZD/ha) at milk price of NZD 6.5/kg MS was 10% greater for HIRG than LIRG + PL and LIRG, and similar (<1.5% numerical difference) between LIRG + PL and LIRG. The average estimated annual N leaching loss from the LIRG and LIRG + PL was 31% and 56% less than the loss from the HIRG. These large reductions in N leaching loss were achieved without a large decrease in profitability (i.e., LIRG and LIRG + PL compared to HIRG). In addition, the estimated reduction in N losses from the LIRG + PL system compared to LIRG suggests that an Italian ryegrass + plantain-based dairy system is a viable strategy to reduce the environmental footprint while maintaining farm profitability. However, the environmental benefits of plantain and Italian ryegrass estimated in this study require further confirmation through direct measurements at full farm level.

Will rising atmospheric CO2 concentration inhibit nitrate assimilation in shoots but enhance it in roots of C3 plants?

Bloom et al. proposed that rising atmospheric CO2 concentrations 'inhibit malate production in chloroplasts and thus impede assimilation of nitrate into protein of C3 plants, a phenomenon that will strongly influence primary productivity and food security under the environmental conditions anticipated during the next few decades'. Previously we argued that the weight of evidence in the literature indicated that elevated atmospheric [CO2 ] does not inhibit NO3 - assimilation in C3 plants. New data for common bean (Phaseolus vulgaris) and wheat (Triticum aestivum) were presented that supported this view and indicated that the effects of elevated atmospheric [CO2 ] on nitrogen (N) assimilation and growth of C3 vascular plants were similar regardless of the form of N assimilated. Bloom et al. strongly criticised the arguments presented in Andrews et al. Here we respond to these criticisms and again conclude that the available data indicate that elevated atmospheric [CO2 ] does not inhibit NO3 - assimilation of C3 plants. Measurement of the partitioning of NO3 - assimilation between root and shoot of C3 species under different NO3 - supply, at ambient and elevated CO2 would determine if their NO3 - assimilation is inhibited in shoots but enhanced in roots at elevated atmospheric CO2 .

Elevated CO2 effects on nitrogen assimilation and growth of C3 vascular plants are similar regardless of N-form assimilated.

Atmospheric carbon dioxide concentration ([CO2]) increased from around 280 ppm in 1750 to 400 ppm in 2016 and is likely to continue to increase throughout this century. It has been argued that wheat, Arabidopsis, and C3 plants in general respond more positively to elevated atmospheric [CO2] under ammonium (NH4+) nutrition than under nitrate (NO3-) nutrition because elevated CO2 inhibits their photoreduction of NO3- and hence reduces their total plant nitrogen (N) assimilation and ultimately growth. Here, it is argued that the weight of evidence in the literature indicates that elevated atmospheric [CO2] does not inhibit NO3- assimilation and growth of C3 vascular plants. New data for common bean and wheat support this view and indicate that the effects of elevated atmospheric [CO2] on N assimilation and growth of C3 vascular plants will be similar regardless of the form of N assimilated.

Horizontal Transfer of Symbiosis Genes within and Between Rhizobial Genera: Occurrence and Importance.

Rhizobial symbiosis genes are often carried on symbiotic islands or plasmids that can be transferred (horizontal transfer) between different bacterial species. Symbiosis genes involved in horizontal transfer have different phylogenies with respect to the core genome of their ‘host’. Here, the literature on legume⁻rhizobium symbioses in field soils was reviewed, and cases of phylogenetic incongruence between rhizobium core and symbiosis genes were collated. The occurrence and importance of horizontal transfer of rhizobial symbiosis genes within and between bacterial genera were assessed. Horizontal transfer of symbiosis genes between rhizobial strains is of common occurrence, is widespread geographically, is not restricted to specific rhizobial genera, and occurs within and between rhizobial genera. The transfer of symbiosis genes to bacteria adapted to local soil conditions can allow these bacteria to become rhizobial symbionts of previously incompatible legumes growing in these soils. This, in turn, will have consequences for the growth, life history, and biogeography of the legume species involved, which provides a critical ecological link connecting the horizontal transfer of symbiosis genes between rhizobial bacteria in the soil to the above-ground floral biodiversity and vegetation community structure.

Effect of elevated temperature on membrane lipid saturation in Antarctic notothenioid fish.

Homeoviscous adaptation (HVA) is a key cellular response by which fish protect their membranes against thermal stress. We investigated evolutionary HVA (long time scale) in Antarctic and non-Antarctic fish. Membrane lipid composition was determined for four Perciformes fish: two closely related Antarctic notothenioid species (Trematomus bernacchii and Pagothenia borchgrevinki); a diversified related notothenioid Antarctic icefish (Chionodraco hamatus); and a New Zealand species (Notolabrus celidotus). The membrane lipid compositions were consistent across the three Antarctic species and these were significantly different from that of the New Zealand species. Furthermore, acclimatory HVA (short time periods with seasonal changes) was investigated to determine whether stenothermal Antarctic fish, which evolved in the cold, stable environment of the Southern Ocean, have lost the acclimatory capacity to modulate their membrane saturation states, making them vulnerable to anthropogenic global warming. We compared liver membrane lipid composition in two closely related Antarctic fish species acclimated at 0 °C (control temperature), 4 °C for a period of 14 days in T. bernacchii and 28 days for P. borchgrevinki, and 6 °C for 7 days in both species. Thermal acclimation at 4 °C did not result in changed membrane saturation states in either Antarctic species. Despite this, membrane functions were not compromised, as indicated by declining serum osmolality, implying positive compensation by enhanced hypo-osmoregulation. Increasing the temperature to 6 °C did not change the membrane lipids of P. borchgrevinki. However, in T. bernacchii, thermal acclimation at 6 °C resulted in an increase of membrane saturated fatty acids and a decline in unsaturated fatty acids. This is the first study to show a homeoviscous response to higher temperatures in an Antarctic fish, although for only one of the two species examined.

Extraction of Pathogenesis-Related Proteins and Phenolics in Sauvignon Blanc as Affected by Grape Harvesting and Processing Conditions.

Thaumatin-like proteins (TLPs) and chitinases are the two main groups of pathogenesis-related (PR) proteins found in wine that cause protein haze formation. Previous studies have found that phenolics are also involved in protein haze formation. In this study, Sauvignon Blanc grapes were harvested and processed in two vintages (2011 and 2012) by three different treatments: (1) hand harvesting with whole bunch press (H-WB); (2) hand harvesting with destem/crush and 3 h skin contact (H-DC-3); and (3) machine harvesting with destem/crush and 3 h skin contact (M-DC-3). The juices were collected at three pressure levels (0.4 MPa, 0.8 MPa and 1.6 MPa), some juices were fermented in 750 mL of wine bottles to determine the bentonite requirement for the resulting wines. Results showed juices of M-DC-3 had significantly lower concentration of proteins, including PR proteins, compared to those of H-DC-3, likely due to the greater juice yield of M-DC-3 and interactions between proteins and phenolics. Juices from the 0.8-1.6 MPa pressure and resultant wines had the highest concentration of phenolics but the lowest concentration of TLPs. This supported the view that TLPs are released at low pressure as they are mainly present in grape pulp but additional extraction of phenolics largely present in skin occurs at higher pressing pressure. Wine protein stability tests showed a positive linear correlation between bentonite requirement and the concentration of chitinases, indicating the possibility of predicting bentonite requirement by quantification of chitinases. This study contributes to an improved understanding of extraction of haze-forming PR proteins and phenolics that can influence bentonite requirement for protein stabilization.

Effect of parity, milking time and stage of lactation on milk yield of Jiangyue donkey (Equus asinus) in North West China.

The study reported in this Research Communication was carried out to examine how parity and milking time affected donkey milk yield under a typical Chinese production system. Eighteen Jiangyue breed donkeys with good health condition, aged between 6 and 9 years, and with parities 3 and 4, were selected. Milk production was measured commencing from the fourth week post-foaling, with a total data collection of 170 d in milk (lactation length). Mean milk yield of the donkeys used in this study was 3·0 kg/donkey/d. Results showed milk yield decreased with days in milk from 3·3 kg/donkey/d at the start of the study, to 2·2 kg/donkey/d by the end of the 170 d. Parity 3 donkeys produced 22% more milk than parity 4 donkeys (3·3 kg/donkey/d vs 2·7 kg/donkey/d). The information provided by this study should aid producers in estimating the expected milk yields from individual donkey, and better predict milk yield over the course of a production cycle.

Aphid-induced accumulation of trehalose in Arabidopsis thaliana is systemic and dependent upon aphid density.

Trehalose is a disaccharide sugar that is now considered to be widely distributed among higher plants. Trehalose has been attributed a number of roles, including control of basic plant processes, such as photosynthesis, and conferring tolerance to abiotic stresses, such as desiccation and high salinity. Trehalose is also a common storage sugar used by insects. In this study, we used laboratory investigations to examine various aspects of trehalose dynamics in an aphid-host plant system (Arabidopsis and the peach potato aphid, Myzus persicae). Trehalose concentrations were measured by [1-H]-NMR. Myzus persicae reared on Arabidopsis, but not on black mustard or spring cabbage, contained considerable quantities of trehalose (5 % w/w dry matter). In Arabidopsis foliage, feeding by aphids induced a density-dependent accumulation of trehalose up to 5 mg g(-1) dry weight. Leaves that were not challenged directly by aphids also exhibited increased trehalose concentrations, indicating that this accumulation was systemic. Trehalose was measured at high concentrations in the phloem sap of plants challenged by aphids, suggesting that aphid feeding induced the plant to produce significant quantities of trehalose, which moved through the plant and into the aphids via the phloem sap. Trehalose was also excreted in the aphid honeydew. Further work is required to clarify whether this trehalose accumulation in Arabidopsis has a direct role or a signalling function in plant tolerance of, or resistance to, aphid feeding, and if a similar accumulation of this sugar occurs when other species or genotypes of aphids are reared on this host plant.

Conditional facilitation of an aphid vector, Acyrthosiphon pisum, by the plant pathogen, pea enation mosaic virus.

Plant pathogens can induce symptoms that affect the performance of insect herbivores utilizing the same host plant. Previous studies examining the effects of infection of tic bean, Vicia faba L. (Fabales: Fabaceae), by pea enation mosaic virus (PEMV), an important disease of legume crops, indicated there were no changes in the growth and reproductive rate of its primary vector the pea aphid, Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). Here, we report the results of laboratory experiments investigating how A. pisum responded to PEMV infection of a different host plant, Pisum sativum L., at different stages of symptom development. Aphid growth rate was negatively related to the age of the host plant, but when they were introduced onto older plants with well-developed PEMV symptoms they exhibited a higher growth rate compared to those developing on uninfected plants of the same age. In choice tests using leaf discs A. pisum showed a strong preference for discs from PEMV-infected peas, probably in response to visual cues from the yellowed and mottled infected leaves. When adults were crowded onto leaves using clip-cages they produced more winged progeny on PEMV-infected plants. The results indicate that PEMV produces symptoms in the host plant that can enhance the performance of A. pisum as a vector, modify the production of winged progeny and affect their spatial distribution. The findings provide further evidence that some insect vector/plant pathogen interactions could be regarded as mutualistic rather than commensal when certain conditions regarding the age, stage of infection and species of host plant are met.

The RAP1 gene confers effective, race-specific resistance to the pea aphid in Medicago truncatula independent of the hypersensitive reaction.

Plant resistance to pathogens is commonly associated with a hypersensitive response (HR), but the degree to which the HR is responsible for incompatibility is subject to debate. Resistance to aphids is likely to share features with resistance to pathogens but is less well understood. Here, we report effective resistance to the pea aphid Acyrthosiphon pisum in Medicago truncatula. Aphids lost weight and died rapidly (within two days) on the resistant genotype Jemalong, which developed necrotic lesions following infestation. Lesions were induced by nonvascular intracellular stylet punctures by aphids, remained localized to the site of stylet entry, stained for the presence of reactive oxygen species, and were similar to the HR induced by the bacterial pathogen Pseudomonas syringae pv. phaseolicola. The implication that aphid-induced lesions confer resistance was tested by quantitative trait loci analysis using recombinant inbred lines derived from a cross between Jemalong and the susceptible genotype DZA315.16. One major locus, RAP1, was identified that was sufficient to confer race-specific resistance against the pea aphid and was mapped to the middle of chromosome 3. Surprisingly, a separate locus, mapping to the top of chromosome 3, governed aphid-induced HR, indicating that the HR-like lesions are not required for RAP1-mediated aphid resistance.

Complex interactions between a plant pathogen and insect parasitoid via the shared vector-host: consequences for host plant infection.

Plant viruses modify the development of their aphid vectors by inducing physiological changes in the shared host plant. The performance of hymenopterous parasitoids exploiting these aphids can also be modified by the presence of the plant pathogen. We used laboratory and glasshouse microcosms containing beans (Vicia faba) as the host plant to examine the interactions between a plant virus (pea enation mosaic virus; PEMV) and a hymenopterous parasitoid (Aphidius ervi) that share the aphid vector/host Acyrthosiphon pisum. Neither PEMV-infection of V. faba, nor the carriage of PEMV virions by A. pisum, affected the growth or morphology of the aphid, or the oviposition behaviour and development of A. ervi. The presence of developing Aphidius ervi larvae within Acyrthosiphon pisum did not affect the ability of the aphids to transmit PEMV. However, by reducing their longevity, parasitism ultimately decreased the time viruliferous aphids were able to inoculate plants. In terms of virus dispersal, parasitized aphids exhibited more movement around experimental arenas than unparasitized controls, causing a slight increase in the proportion of beans infected with PEMV. Exposure to adult Aphidius ervi caused Acyrthosiphon pisum to rapidly drop off bean plants and disperse to new hosts, resulting in considerably higher plant infection rates (70%) than that seen in control arenas (25%). The results of this investigation demonstrate that when parasitoids are added to a plant-pathogen-vector system, benefits to the host plant due to reduced herbivore infestation must be balanced against the consequences of parasitoid-induced aphid dispersal and a subsequent increase in the level of plant infection.