Anna Lehrman

Estimating direct resistance in willows against a major insect pest, Phratora vulgatissima, by comparing life history traits

Many Salix species (Salicaceae) have a high energy quotum and efficient nitrogen use, resulting in a high and stable biomass production, making these plants suitable as bioenergy crops. However, Salix coppices are sensitive to herbivory by chrysomelid beetles, such as Phratora vulgatissima L. (Coleoptera: Chrysomelidae), and to maintain high biomass yields over several years, durable plant resistance is necessary. We have developed a reliable and efficient bioassay that can be used as a tool for detection of resistance against P. vulgatissima in Salix. Screening of six clones of one susceptible (Salix viminalis L.) and one resistant (Salix dasyclados Wimm.) willow species, by the response of several life history traits of larval and adult P. vulgatissima, identified egg production as the most reliable trait for resistance. We show that a 2‐week bioassay of oviposition rate is sufficient for a trustworthy estimation of both total egg production and survival of adult leaf beetles. In addition, to elucidate the variation in resistance among the studied clones, leaf concentrations of various phenolic compounds were compared with the key life history traits of the beetles. Phenolic compounds, especially salicylates, have previously been reported as the main factor controlling Salix resistance against P. vulgatissima. Indeed we found higher levels of salicylates in the resistant clones and larval performance was negatively correlated to the concentration of salicylates in the leaves. In addition, we found indications of negative effects of a luteolin derivative and quercetin‐3‐glucoside on leaf beetle performance, but further studies are needed to elucidate the specific roles that the different phenolic compounds play in plant resistance.

Host-plant genotype mediates supply and demand of animal food in an omnivorous insect

1. Omnivorous predators can protect plants from herbivores, but may also consume plant material themselves. Omnivores and their purely herbivorous prey have previously been thought to respond similarly to host‐plant quality. However, different responses of omnivores and herbivores to their shared host plants may influence the fitness, trophic identity, and population dynamics of the omnivores.

Influence of pea lectin expressed transgenically in oilseed rape (Brassica napus) on adult pollen beetle (Meligethes aeneus)

Adult pollen beetles were exposed to three transgenic lines of Brassica napus L. expressing pea lectin up to 0.7% of total soluble protein in anthers. Two lines not expressing the pea lectin were used as controls. Adult responses were recorded in two experiments where beetles were fed either racemes or anthers. There was no effect of plant line on feeding rate, weight change or oviposition rate. A slightly reduced egg size was recorded when beetles were feeding on one of the plant lines with high concentration of pea lectin. Previous studies on the effect of pea lectin on pollen beetles have only focused on larval performance. The results of this study suggest that pea lectin has no adverse effect on the pest in its most damaging life stage.

Does pea lectin expressed transgenically in oilseed rape (Brassica napus) influence honey bee (Apis mellifera) larvae

The European honey bee (Apis mellifera) is important both for pollination and for honey production. Pollen is the major protein source for bees, which exposes them directly to changes in pollen quality e.g. through genetic engineering. In order to create a worst case scenario regarding pea lectin (PSL) expressed transgenically in oilseed rape anthers and pollen, the maximum amount of dried pollen that could be mixed in an artificial diet without negatively affecting larval performance (1.5% w/w) was fed to bee larvae. Pollen from two transgenic plant lines expressing PSL up to 1.2% of total soluble protein and pollen from one non-transgenic line was added to the same diet and used as a pollen control. When these three pollen diets and the control diet (without added pollen) were compared, no negative effect from the pollen of the transgenic plants could be detected on larval mortality, weight, or development time. An increased weight and a reduced developmental time were recorded for larvae on all diets containing pollen when compared to the diet without pollen.

Constitutive and herbivore-induced systemic volatiles differentially attract an omnivorous biocontrol agent to contrasting Salix clones

Little is known about how herbivore-induced plant volatiles affect omnivorous predators. Here we show that the key predator Anthocoris nemorum is differentially attracted to three Salix clones when these are damaged by the detrimental blue willow beetle (Phratora vulgatissima). At least two volatile plant compounds were induced by the herbivore, and these were antennal active in the predator. The results elucidate how plants may recruit omnivorous predators when damaged. These findings could be utilized in crop breeding for increased resistance against herbivores.

Impact of herbivory and pollination on performance and competitive ability of oilseed rape transformed for pollen beetle resistance

Competitive ability of transgenic oilseed rape transformed with a pea lectin gene was estimated by comparisons of its performance when grown in a mixture with its non-transgenic counterpart and when grown singly, with and without pollen beetles present. The experiments were carried out in cages, once with bumblebees as pollinators and once without. In the absence of herbivory but with the presence of bumblebees, singly grown plant lines without lectin generally performed better than lines with lectin. Pollen beetles affected plant growth and reproduction, but there were no consistent differences between the lectin and non-lectin plant lines indicating that the transgenic trait did not protect plants from pest attack. Herbivory reduced the number of seeds when bumblebees were present. In the absence of bumblebees, however, plants produced more seeds with pollen beetles than without, indicating that some pollination was carried out by the beetles. Efficient pollination affected the competitive abilities of the lines; lectin lines were more competitive with bumblebees present and the reverse was true when bumblebees were absent. In the presence of bumblebees, lectin lines gained from being grown mixed with its non-transgenic counterpart. Because the transgenic plants expressed pea lectin in developing pollen it is possible that pollen quality in those plants was reduced, which may explain why the lectin lines had an advantage over non-lectin lines when exchange of pollen between the two plant types was facilitated by bumblebees.

Performance of an Herbivorous Leaf Beetle (Phratora vulgatissima) on Salix F2 Hybrids: the Importance of Phenolics

The genotype of the plant determines, through the expression of the phenotype, how well it is suited as food for herbivores. Since hybridization often results in profound genomic alterations with subsequent changes in phenotypic traits, it has the potential to significantly affect plant-herbivore interactions. In this study, we used a population of F2 hybrids that originated from a cross between a Salix viminalis and a Salix dasyclados genotype, which differed in both phenolic content and resistance to the herbivorous leaf beetle Phratora vulgatissima. We screened for plants that showed a great variability in leaf beetle performance (i.e., oviposition and survival). By correlating leaf phenolics to the response of the herbivores, we evaluated the importance of different phenolic compounds for Salix resistance to the targeted insect species. The performance of P. vulgatissima varied among the F2 hybrids, and two patterns of resistance emerged: leaf beetle oviposition was intermediate on the F2 hybrids compared to the parental genotypes, whereas leaf beetle survival demonstrated similarities to one of the parents. The findings indicate that these life history traits are controlled by different resistance mechanisms that are inherited differently in the hybrids. Salicylates and a methylated luteolin derivative seem to play major roles in hybrid resistance to Phratora vulgatissima. Synergistic effects of these compounds, as well as potential threshold concentrations, are plausible. In addition, we found considerable variation in both distributions and concentrations of different phenolics in the F2 hybrids. The phenolic profiles of parental genotypes and F2 hybrids differed significantly (e.g., novel compounds appeared in the hybrids) suggesting genomic alterations with subsequent changes in biosynthetic pathways in the hybrids.

Effect of pea lectin expressed transgenically in oilseed rape on pollen beetle life‐history parameters

When investigating resistance to herbivorous insects in transgenic plants, it is primarily the damaging stage of the insect that is targeted. In our study, we showed that small effects of the transgenic plant on the pest performance might lead to a clear effect on the pest population level. Pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), larvae were reared on three transgenic lines of Brassica napus L. (Brassicaceae) expressing pea (Pisum sativum L.) lectin at 0.2, 0.6, and 0.7% of total soluble protein in anthers, and then monitored until the adult stage. Two lines without the gene construct and consequently containing no pea lectin were used as controls. The egg viability, larval weight, development time, and survival rate were recorded. In a separate experiment, the development time from egg to adult, adult weight, and winter survival of pollen beetles developed on intact plants were recorded. Survival of larvae to adult stage was significantly lower on the transgenic plant lines, independent of lectin level. Survival during hibernation was only 2.4% and was not affected by plant line. Combining previously published results from adult feeding and oviposition assays with the new data on larval development we summarize the effect of the transgenic oilseed rape on the pollen beetle, from oviposition in spring to overwintered adults of the new generation. Net reproductive rate was then shown to be reduced by half on the transgenic oilseed rape compared to the control plants.