Effects of aphid parasitism on host plant fitness in an aphid-host relationship.

Aphids are serious agricultural insect pests which exploit the phloem sap of host plants and thus transmit pathogens to their hosts. However, the degree to which aphid parsitism affects the fitness of the host plants is not well understood. The aphid, Macrosiphoniella yomogicola, parasitizes the mugwort Artemisia montana in Japan. During summer most mugworts carry aphids, but most aphid colonies die out after the budding of A. montana inflorescences in late summer. A few aphid colonies survive to late autumn, at which point sexuparae appear to later lay overwintering eggs after copulation. The death of the aphid colonies seems to be caused by biochemical changes in the phloem sap in the host plant coincident with the budding of inflorescences. The surviving aphid colonies may suppress the budding of inflorescences to allow persistence of their genetic line into the following year. Our investigations demonstrate that aphid parasitism did not affect host plant growth, but that it did significantly decrease the number of inflorescences and the average weight of floral buds. Our results indicate that aphid parasitism has a strong negative effect on the fitness of host plants. The manner in which the aphids suppress floral budding in their hosts is worth examining from the perspective of the evolution of aphid-plant interactions.

Salicylic acid-induced changes in physiological parameters and genes of the flavonoid biosynthesis pathway in Artemisia vulgaris and Dendranthema nankingense during aphid feeding.

Phloem-feeding aphids cause serious damage to plants. The mechanisms of plant-aphid interactions are only partially understood and involve multiple pathways, including phytohormones. In order to investigate whether salicylic acid (SA) is involved and how it plays a part in the defense response to the aphid Macrosiphoniella sanbourni, physiological changes and gene expression profiles in response to aphid inoculation with or without SA pretreatment were compared between the aphid-resistant Artemisia vulgaris 'Variegata' and the susceptible chrysanthemum, Dendranthema nankingense. Changes in levels of reactive oxygen species, malondialdehyde (MDA), and flavonoids, and in the expression of genes involved in flavonoid biosynthesis, including PAL (phenylalanine ammonia-lyase), CHS (chalcone synthase), CHI (chalcone isomerase), F3H (flavanone 3-hydroxylase), F3'H (flavanone 3'-hydroxylase), and DFR (dihydroflavonol reductase), were investigated. Levels of hydrogen peroxide, superoxide anions, MDA, and flavonoids, and their related gene expression, increased after aphid infestation and SA pretreatment followed by aphid infestation; the aphid-resistant A. vulgaris exhibited a more rapid response than the aphid-susceptible D. nankingense to SA treatment and aphid infestation. Taken together, our results suggest that SA could be used to increase aphid resistance in the chrysanthemum.

Biology and morphology of immature stages of Adosomus roridus (Coleoptera: Curculionidae: Lixinae).

Mature larva and pupa of Adosomus (s. str.) roridus (Pallas, 1781) (Curculionidae: Lixinae: Cleonini) are described and compared with ten other taxa of Cleonini with known larvae. This weevil is an oligophagous species on the Asteraceae family. From our observations in Slovakia, we found active adults from April to September in dry sunny places within extensively used or fallow vineyards and in ruderal vegetation with host plants. The weevil is a root borer--larvae, pupae and fresh adults were collected from the root necks and roots of Common Tansy (Tanacetum vulgare L.) and rarely from Mugwort (Artemisia vulgaris L.). Each plant was usually occupied by one larva, or more rarely with two or three larvae. The new generation of adult individuals appeared from early summer to autumn. Both larvae and some of the adults overwinter, which is quite unique within Cleonini.

Spatial and temporal dynamics of Aroga moth (Lepidoptera: Gelechiidae) populations and damage to sagebrush in shrub steppe across varying elevation.

Spatial and temporal variation in the density of the Aroga moth, Aroga websteri Clarke (Lepidoptera: Gelechiidae), and in its damage to its host plant, big sagebrush (Artemisia tridentata Nuttall), were examined at 38 sites across a shrub steppe landscape in mountain foothills of northern Utah. Sites were sampled from 2008 to 2012 during and after an outbreak of the moth, to assess whether and how local variation in moth abundance, survivorship, and damage to the host plant was accounted for by sagebrush cover, elevation, slope, aspect, or incident solar radiation. As moth numbers declined from a peak in 2009, individual sites had a consistent tendency in subsequent years to support more or fewer defoliator larvae. Local moth abundance was not correlated with sagebrush cover, which declined with elevation, and moth survivorship was highest at intermediate elevations (1,800-2,000 m). North-facing stands of sagebrush, characterized by lower values of incident solar radiation, were found to be especially suitable local habitats for the Aroga moth, as reflected in measures of both abundance and feeding damage. This high habitat suitability may result from favorable microclimate, both in its direct effects on the Aroga moth and in indirect effects through associated vegetative responses. North-facing sites also supported taller and more voluminous sagebrush plants in comparison to south-facing sites. Thus, the moth is reasonably predictable in the sites at which it is likely to occur in greatest numbers, and such sites may be those that in fact have most potential to recover from feeding damage.

Comparative study of the volatiles' composition of healthy and larvae-infested Artemisia ordosica.

Volatiles emitted by healthy Artemisia ordosica (Asteraceae) and plants infested with larvae of Sphenoptera sp. (Coleoptera: Buprestidae) or Holcocerus artemisiae (Lepidoptera: Cossidae) were obtained using a dynamic headspace method and analysed by automatic thermal desorption/gas chromatography/mass spectrometry (ATD/GC/MS). Twenty-eight major compounds were identified, and qualitative and quantitative differences were compared. The novel green leaf volatiles 2-hexenal, (Z)-3-hexen-1-ol, 2-hexen-1-ol 1-hexanol, and (Z)-3-hexen-1-ol acetate, the terpenoids alpha-copaene, beta-cedrene, and (E,E)-alpha-farnesene, and the ester methyl salicylate were present in all infested plants. Volatiles from healthy plants were dominated by D-limonene (32.14%), beta-pinene (16.63%), beta-phellandrene (16.06%), and sabinene (12.88%). Volatiles from Sphenoptera sp. larvae-infested plants were dominated by D-limonene (24.74%), beta-pinene (21.05%), alpha-pinene (19.39%), and sabinene (11.64%), whereas volatiles from H. artemisiae larvae-infested plants were dominated by D-limonene (31.76%), sabinene (18.49%), ocimene (15.93%), and beta-phellandrene (10.59%). In addition to the qualitative variation, a larvae-induced quantitative change in the proportion of terpenoids in the blends was also a noticeable feature.

Scanning the European corn borer (Ostrinia spp.) genome for adaptive divergence between host-affiliated sibling species.

It has recently been shown that the European corn borer, a major pest of maize crops, is actually composed of two genetically differentiated and reproductively isolated taxa, which are found in sympatry over a wide geographical range in Eurasia. Each taxon is adapted to specific host plants: Ostrinia nubilalis feeds mainly on maize, while O. scapulalis feeds mainly on hop or mugwort. Here, we present a genome scan approach as a first step towards an integrated molecular analysis of the adaptive genomic divergence between O. nubilalis and O. scapulalis. We analysed 609 AFLP marker loci in replicate samples of sympatric populations of Ostrinia spp. collected on maize, hop and mugwort, in France. Using two genome scan methods based on the analysis of population differentiation, we found a set of 28 outlier loci that departed from the neutral expectation in one or the other method (of which a subset of 14 loci were common to both methods), which showed a significantly increased differentiation between O. nubilalis and O. scapulalis, when compared to the rest of the genome. A subset of 12 outlier loci were sequenced, of which 7 were successfully re-amplified as target candidate loci. Three of these showed homology with annotated lepidopteran sequences from public nucleotide databases.

An air transfer experiment confirms the role of volatile cues in communication between plants.

Previous studies reported that sagebrush plants near experimentally clipped neighbors experienced less herbivory than did plants near unclipped neighbors. Blocking air flow with plastic bags made this effect undetectable. However, some scientists remained skeptical about the possibility of volatile communication between plants since the existence and identity of a cue that operates in nature have never been demonstrated. We conducted an air transfer experiment that collected air from the headspace of an experimentally clipped donor plant and delivered it to the headspace of an unclipped assay plant. We found that assay plants treated with air from clipped donors were less likely to be damaged by naturally occurring herbivores in a field experiment. This simple air transfer experiment fulfills the most critical of Koch's postulates and provides more definitive evidence for volatile communication between plants. It also provides an inexpensive experimental protocol that can be used to screen plants for interplant communication in the field.

Did the introduction of maize into Europe provide enemy-free space to Ostrinia nubilalis? Parasitism differences between two sibling species of the genus Ostrinia.

We examined whether maize offers enemy-free space (EFS) to its pest Ostrinia nubilalis, and may thereby have contributed to its divergence from the sibling species, Ostrinia scapulalis, feeding mainly on mugwort, when introduced into Europe five centuries ago. We collected Ostrinia larvae on maize (70 populations, 8425 individuals) and mugwort (10 populations, 1184 individuals) and recorded parasitism using both traditional (counting emerging parasitoids) and molecular methods (detection by specific polymerase chain reaction). The main parasitoid was Macrocentrus cingulum (Braconidae). On mugwort, parasitism was twice that on maize, and parasitoid-related mortality was 8 times higher. This suggests that maize affords substantial EFS to Ostrinia feeding on it. The lower Mortality:Infestation ratio in maize suggests that O. nubilalis' immune response might be stronger than that of O. scapulalis. If so, adapting to maize and diverging from O. scapulalis would decrease the impact of parasitism on O. nubilalis at both ecological and evolutionary levels.

Sympatric host races of the European corn borer: adaptation to host plants and hybrid performance.

The European corn borer (ECB), Ostrinia nubilalis, is a major pest of maize crops. In Europe, two sympatric host races are found: one feeds on maize (Zea mays) and the other mainly on mugwort (Artemisia vulgaris). The two host races are genetically differentiated, seldom crossing in the laboratory or in the field, and females preferentially lay eggs on their native host species. We conducted two independent experiments, in field and greenhouse conditions, to determine whether the two host races are locally adapted to their host species. The effect of larval density and the performance of hybrids were also investigated. Despite some differences in overall larval feeding performance, both experiments revealed consistent patterns of local adaptation for survival and for larval weight in males. In females the same trend was observed but with weaker statistical support. F1 hybrids did not seem to be disadvantaged compared with the two parental races. Overall, our results showed that both host races are physiologically adapted to their native host. The fitness trade-off between the two host plants provides a potential driving force for ecological speciation in this species.

Short- and longer-term effects of fire and herbivory on sagebrush communities in south-central Montana.

To better understand the role of herbivory and fire as potential disturbance processes in sagebrush communities, we examined responses of a grazing ungulate, elk (Cervus elaphus), following prescribed burning of sagebrush (Artemisia tridentata ssp. vaseyana) in south-central Montana (USA.) with concurrent monitoring of changes in plant production, nutritional quality, and community diversity from 1989-1999. Burning transformed low-diversity, sagebrush-dominated communities into high-diversity, graminoid-forb communities that persisted for 10 years without significant reestablishment of sagebrush. Elk increased use of burned sites one year after burning, but elk use returned to pre-burn levels over the next two to nine years. Forage biomass and nutritional quality declined after initial increases that coincided with increased elk use. Increases in elk use appeared to be influenced by increases in combined graminoid and forb production and changes in structural vegetation characteristics that permitted greater foraging efficiency. Declines in use were associated with loss of nutritional enhancement and declines in combined graminoid and forb production. Managers may observe only short-term responses from grazing ungulates to prescribed fire in sagebrush communities, but can expect longer-term increases in plant diversity and establishment of graminoid-forb communities.

Priming of plant defense responses in nature by airborne signaling between Artemisia tridentata and Nicotiana attenuata.

Plants release volatile organic compounds (VOCs) in response to wounding and herbivore attack, some of which trigger responses in neighboring unattacked plants in the laboratory under conditions that are not likely to occur in the real world. Whether plants 'eavesdrop' on the volatile emissions of their neighbors in nature is not known. The best documented field study of between-species signaling via above-ground VOCs involves increases in fitness parameters of native tobacco (Nicotiana attenuata) transplanted adjacent to clipped sagebrush (Artemesia tridentata tridentata). Clipped sagebrush releases many biologically active VOCs, including methyl jasmonate (MeJA), methacrolein and a series of terpenoid and green leaf VOCs, of which MeJA, while active under laboratory conditions, is not released in sufficient quantities to directly elicit induced resistance in the field. Here we demonstrate, with laboratory and field-based experiments, that priming (rather than direct elicitation) of native N. attenuata's induced chemical defenses by a sagebrush-released VOC bouquet can account for earlier findings. With microarrays enriched in N. attenuata herbivore-regulated genes, we found transcriptional responses in tobacco growing adjacent to clipped sagebrush foliage, but failed to detect the direct elicitation of defensive chemicals or proteins. However, we observed an accelerated production of trypsin proteinase inhibitors when Manduca sexta caterpillars fed on plants previously exposed to clipped sagebrush. This readying of a defense response, termed priming, results in lower total herbivore damage to plants exposed to clipped sagebrush and in a higher mortality rate of young Manduca caterpillars. Our study demonstrates priming of plant defense responses as a mechanism of plant-plant signaling in nature, and provides an example for the analysis of between-plant signaling under ecologically realistic conditions. Although we describe priming as a potential mechanism for signaling between plants in nature, we critically discuss the ecological relevance of the particular interaction.

Laboratory evaluation of Artemisia annua L. extract and artemisinin activity against Epilachna paenulata and Spodoptera eridania.

Ethanolic extract of aerial parts of Artemisia annua L. and artemisinin were evaluated as anti-insect products. In a feeding deterrence assay on Epilachna paenulata Germ (Coleoptera: Coccinellidae) larvae, complete feeding rejection was observed at an extract concentration of 1.5 mg/cm2 on pumpkin leaf tissue. The same concentration produced a feeding inhibition of 87% in Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae). In a no-choice assay, both species ate less and gained less weight when fed on leaves treated with the extract. Complete mortality in E. paenulata and 50% mortality in S. eridania were observed with extract at 1.5 mg/cm2. Artemisinin exhibited a moderate antifeedant effect on E. paenulata and S. eridania at 0.03-0.375 mg/cm2. However, a strong effect on survival and body weight was observed when E. paenulata larvae were forced to feed on leaves treated at 0.03 and 0.075 mg/cm2. Artemisia annua ethanolic extract of aerial parts at 1.5 mg/cm2 showed no phytotoxic effect on pumpkin seedlings.

Genetic isolation between two sympatric host plant races of the European corn borer, Ostrinia nubilalis Hubner. II: assortative mating and host-plant preferences for oviposition.

The European corn borer, Ostrinia nubilalis Hubner, colonized maize (Zea mays L.) after its introduction into Europe about 500 years ago and is now considered one of the main pests of this crop. In northern France, two sympatric host races have been described: one feeding on maize and the other on mugwort (Artemisia vulgaris L.) and hop (Humulus lupulus L.). In a previous study, we showed that mating between the two races may be impeded by differences in the timing of moth emergence and in the composition of the sex pheromone produced by the females. In this study, we further investigated the genetic isolation of these two races using strains from the maize (Z strain) and mugwort (E strain) races selected for diagnostic alleles at two allozyme loci. In a cage containing maize and mugwort plants and located in natural conditions, mating between individuals of the same strain occurred more often than mating between males and females of the E and Z strains. In particular, we obtained no evidence for crosses between Z females and E males. We also found that females of the Z strain laid their eggs almost exclusively on maize, whereas females of the E strain laid their eggs preferentially, but not exclusively, on mugwort. These results suggest that the genetic differentiation between the two host races may also be favored by host-plant preference, one of the first steps toward sympatric speciation.

Genetic isolation between two sympatric host-plant races of the European corn borer, Ostrinia nubilalis Hübner. I. Sex pheromone, moth emergence timing, and parasitism.

Adaptation to different environments may be a powerful source of genetic differentiation between populations. The biological traits selected in each environment can pleiotropically induce assortative mating between individuals of these genetically differentiated populations. This situation may facilitate sympatric speciation. Successful host shifts in phytophagous insects provide some of the best evidence for the ecological speciation that occurs, or has occurred, in sympatry. The European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae), colonized maize after its introduction into Europe by humans about 500 years ago. In northern France, two sympatric host races feed on maize (Zea mays) and mugwort (Artemisia vulgaris), respectively. We investigated the factors involved in the genetic isolation of these two races at a field site near Paris, France. We identified two biological differences that might make a significant contribution to the genetic divergence between sympatric populations feeding on the two host plants. First, assortative mating may be due to differences in the moth emergence pattern between the two races: mugwort-race moths emerged on average 10 days earlier than maize-race moths. In addition, the males emerged earlier than females in both races. Hence, the likelihood of mating between maize-race males and mugwort-race females was higher than that of mating between mugwort-race males and maize-race females. Second, the females feeding on mugwort and maize produced sex pheromones with different E/Z isomeric ratios of delta-11-tetradecenyl acetate. This difference in mate recognition systems reinforces the potential for assortative mating in the two races. During the experiment, overwintering mortality was much lower on maize than on mugwort. This difference was due to a braconid parasitoid wasp, Macrocentrus cingulum, that killed more than 50% of the larvae overwintering on mugwort but did not infest larvae diapausing on maize. Hence, by colonizing maize, European corn borer populations probably escaped from numerous predators, competitors, and parasitoids, such as M. cingulum. This decrease in host-associated selection may have favored the colonization of this new host. Finally, throughout this experiment we observed selection at two allozyme loci (or at linked loci): Tpi and Mpi. The Tpi locus is tightly linked with the genes involved in the response of the male to the sex pheromone and in developmental timing. The location of these traits on the Z chromosome may play a role in shortening the time required for the evolution of premating barriers.

Host-plant-associated genetic differentiation in Northern French populations of the European corn borer.

The phytophagous insects that damage crops are often polyphagous, feeding on several types of crop and on weeds. The refuges constituted by noncrop host plants may be useful in managing the evolution in pest species of resistance to the Bacillus thuringiensis toxins produced by transgenic crops. However, the benefits of these refuges may be limited because host-plant diversity may drive genetic divergence and possibly even host-plant-mediated sympatric speciation. The European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), is the main pest of maize in Europe and North America, where it was introduced early in the 20th century. It has a wide host range but feeds principally on mugwort (Artemisia vulgaris L.) and maize (Zea mays L.). O. nubilalis is found on mugwort only in the northern part of France, whereas it is found on maize throughout France. The extent of genetic variation at allozyme markers was investigated in populations collected from the two host plants over the entire geographical distribution of the European corn borer on mugwort in France. Allelic differentiation between pairs of populations and hierarchical analyses of pools of samples from each host plant indicate that the group of populations feeding on maize differed from the group of populations feeding on mugwort. Our results suggest (1) host-plant-related divergent selection at the genomic region surrounding the Mpi locus and (2) limited gene flow between the populations feeding on mugwort and those infesting maize fields. These data indicate that adults emerging from mugwort would not be useful for managing the evolution of resistance to the B. thuringiensis toxins in European corn borer populations.