André Gassmann

The potential for biological control of invasive alien aquatic weeds in Europe: a review

A retrospective analysis shows that invasive, alien, free-floating and emergent aquatic weeds in Europe are good targets for classical biological control, and that genus-specific chrysomelid and curculionid beetles offer the most potential. Ludwigia spp., Azolla filiculoides, Lemna minuta, Crassula helmsii and Hydrocotyle ranunculoides should be prioritised as targets. Fungal pathogens have been under-utilised as classical agents but, whilst they may have some potential against free-floating weeds, they appear to be poor candidates against submerged species, although the suitability of arthropod agents against these difficult targets still merits investigation. The use of indigenous pathogens as inundative agents (mycoherbicides) shows some promise.

Morphological, molecular and biological evidence reveal two cryptic species in Mecinus janthinus Germar (Coleoptera, Curculionidae), a successful biological control agent of Dalmatian toadflax, Linaria dalmatica (Lamiales, Plantaginaceae)

A combined morphological, molecular and biological study shows that the weevil species presently named Mecinus janthinus is actually composed of two different cryptic species: M. janthinus Germar, 1821 and M. janthiniformis Toševski & Caldara sp.n. These species are morphologically distinguishable from each other by a few very subtle morphological characters. On the contrary, they are more readily distinguishable by both molecular and biological characters. A molecular assessment based on the mitochondrial DNA cytochrome oxidase subunit II gene revealed fixed differences between the two species with p‐distances between samples of both species ranging from 1.3 to 2.4%. In addition to this, the larvae of the two species are found to develop on different species within the genus Linaria (Plantaginaceae): M. janthinus is associated with yellow toadflax (L. vulgaris) and M. janthiniformis with broomleaf toadflax (L. genistifolia) and Dalmatian toadflax (L. dalmatica). Molecular and host use records further suggest the occurrence of a third species associated with L. vulgaris within M. janthinus, sampled from north Switzerland, central Hungary and east Serbia. The significance of these new findings is of particular importance because species of the M. janthinus group are used, or are potential candidates, for the biological control of invasive toadflaxes in North America.

Host‐associated genetic differentiation in a seed parasitic weevil Rhinusa antirrhini (Coleptera: Curculionidae) revealed by mitochondrial and nuclear sequence data

Plant feeding insects and the plants they feed upon represent an ecological association that is thought to be a key factor for the diversification of many plant feeding insects, through differential adaptation to different plant selective pressures. While a number of studies have investigated diversification of plant feeding insects above the species level, relatively less attention has been given to patterns of diversification within species, particularly those that also require plants for oviposition and subsequent larval development. In the case of plant feeding insects that also require plant tissues for the completion of their reproductive cycle through larval development, the divergent selective pressure not only acts on adults, but on the full life history of the insect. Here we focus attention on Rhinusa antirrhini (Curculionidae), a species of weevil broadly distributed across Europe that both feeds on, and oviposits and develops within, species of the plant genus Linaria (Plantaginaceae). Using a combination of mtDNA (COII) and nuclear DNA (EF1‐α) sequencing and copulation experiments we assess evidence for host associated genetic differentiation within R. antirrhini. We find substantial genetic variation within this species that is best explained by ecological specialisation on different host plant taxa. This genetic differentiation is most pronounced in the mtDNA marker, with patterns of genetic variation at the nuclear marker suggesting incomplete lineage sorting and/or gene flow between different host plant forms of R. antirrhini, whose origin is estimated to date to the mid‐Pliocene (3.77 Mya; 2.91–4.80 Mya).

Avoiding conflicts between insect and weed biological control: selection of non-target species to assess host specificity of cabbage seedpod weevil parasitoids

Abstract:  Classical biological control of insect pests and weeds may lead to potential conflicts, where insect pests are closely related to weed biological control agents. Such a conflict may occur in the classical biological control of the cabbage seedpod weevil, Ceutorhynchus obstrictus (Marsham) in North America, which belongs to the same subfamily, Ceutorhynchinae, as a number of agents introduced or proposed for introduction against non‐indigenous invasive weed species. We propose a step‐by‐step procedure to select non‐target species and thereby to develop a non‐target species test list for screening candidate entomophagous biological control agents of a herbivore pest insect in a way that would simultaneously evaluate non‐target potential on weed biological control agents and other non‐target species. Using these recommendations, we developed a non‐target test list for host specificity evaluations in the area of origin (Europe) and the area of introduction (North America) for cabbage seedpod weevil parasitoids. Scientifically based predictions on expected host–parasitoid interactions and ecological information about the ecological host range in the area of origin can help avoid conflicts, while still allowing the introduction of safe and effective agents against both insect pests and weeds.

Anatomical injury induced by the eriophyid mite Aceria anthocoptes on the leaves of Cirsium arvense

Anatomical injury of the leaves of the invasive species, Cirsium arvense (L.) Scop., caused by the eriophyid mite Aceria anthocoptes (Nal.), which is the only eriophyid mite that has been recorded on C. arvense worldwide, is described. The injury induced by the mite feeding on the leaves of C. arvense results in visible russeting and bronzing of the leaves. Other conspicuous deformations are folding and distortion of the leaf blade and curling of leaf edge, as well as gradual drying of leaves. The anatomical injury of the mature leaves of field-collected plants was limited to the epidermis of the lower leaf surface. However, on young leaves of experimentally infested plants, rust mite injuries extend to epidermal cells on both leaf surfaces and to those of deeper mesophyll layers. On these leaves, lesions on the lower leaf surface even affected the phloem of the vascular bundles. Leaf damage induced by A. anthocoptes is discussed with regard to the mite’s potential as a biological control agent of C. arvense.

Performance of potential European biological control agents of Vincetoxicum spp. with notes on their distribution.

Exploratory surveys were conducted in Europe to locate potential biological control agents of Vincetoxicum nigrum and Vincetoxicum rossicum and to collect known specialist herbivores of Vincetoxicum hirundinaria (Abrostola asclepiadis, Chrysolina a. asclepiadis, Eumolpus asclepiadeus and Euphranta connexa). Development of the most abundant herbivores found attacking leaves, roots, and developing seeds was evaluated in the laboratory on three Vincetoxicum spp. Field surveys revealed that the highest diversity of herbivores was associated with the abundant and widespread V. hirundinaria. No new herbivores were reported from V. hirundinaria or V. nigrum, but the leaf‐feeding noctuid Hypena opulenta was recorded for the first time attacking V. rossicum and V. scandens in Ukraine. Based upon larval survival and development and adult fecundity, the leaf feeders A. asclepiadis, C. a. asclepiadis, and H. opulenta, perform better on V. nigrum and V. rossicum than on V. hirundinaria. Larval performance of the root feeder E. asclepiadeus followed a similar pattern, but adult fecundity of this insect did not vary among host plant species. Immature development time of the pre‐dispersal seed feeder E. connexa is similar among hosts, but larvae grow larger on V. nigrum and V. hirundinaria than on V. rossicum. All herbivores are promising biological control agents of Vincetoxicum in North America and we have prioritized host range testing with H. opulenta and E. asclepiadeus.

Host-associated genetic divergence and taxonomy in the Rhinusa pilosa Gyllenhal species complex: an integrative approach.

A combined taxonomic, morphological, molecular and biological study revealed that stem‐galling weevils from the genus Rhinusa associated with toadflaxes from the genus Linaria (Plantaginaceae) are composed of three different species: Rhinusa pilosa, Rhinusa brondelii and Rhinusa rara sp.n. The authentic field host plants are respectively, Linaria vulgaris, Linaria purpurea and Linaria genistifolia/ Linaria dalmatica. These weevil species can be distinguished from each other by a few subtle morphological characteristics, mainly in the shape of the rostrum and of the integument. An analysis of the mitochondrial [cytochrome oxidase subunit II gene (COII) and 16S ribosomal RNA gene (16S)] and nuclear (elongation factor‐1α, EF‐1α) sequence data revealed high genetic divergence among these species. Uncorrected pairwise distances on mtCOII gene were 14.3% between R. pilosa and R. brondelii, 15.7% between R. pilosa and R. rara, while R. brondelii and R. rara were approximately 11% divergent from each other. Divergences obtained on 16S and nuclear EF‐1α genes were congruent. However, substantial intraspecific mitochondrial divergence was recorded for all studied populations of R. pilosa s.s. showing two mtDNA lineages, with estimated COII and 16S divergences of 4% and 1.6%, respectively. Nuclear pseudogenes (Numts) and Wolbachia influence, although recorded within both lineages, were excluded as possible causatives of the mtDNA divergence, while EF‐1α indicated absence of lineage sorting. Species from the R. pilosa complex are estimated to have diverged from each other approximately 7.2 million years ago (mya; late Miocene), while R. brondelii and R. rara diverged from each other about 4.7 mya (early Pliocene).

PCR-RFLP-based method for reliable discrimination of cryptic species within Mecinus janthinus species complex (Mecinini, Curculionidae) introduced in North America for biological control of invasive toadflaxes

Several populations of the stem-mining weevil Mecinus janthinus Germar species complex (Mecinini, Curculionidae), identified based on morphological characteristics, have been introduced in North America for the biological control of invasive toadflaxes of European origin: Linaria vulgaris Miller and L. dalmatica (L.) Miller (Plantaginaceae). According to the mitochondrial cytochrome oxidase subunit II (COII) gene haplotype divergence of Mecinus janthinus species complex, a total of 20 M. janthinus s.s., 3 M. janthinus s.l. of the ‘speciosa’ genotype and 29 M. janthiniformis haplotypes have been recorded across their native range in central and southeastern Europe. A polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) diagnostic assay of COII gene using Hpy188III and MnlI enzyme-mix, was developed for fast and cost-effective discrimination of these morphologically very similar cryptic weevil species. It is shown that digestion generates unique 4-fragment restriction profile in M. janthinus s.s., 2-fragment profile in M. janthiniformis and 3-fragment profile in M. janthinus s.l. ‘speciosa’ group of haplotypes, allowing precise identification of each species or genotype. The proposed method represents a practical tool for fast and accurate identification of the target biocontrol agents and should prevent using inappropriate weevil species in redistribution programs for biological control of invasive toadflax species.

Revision of Mecinus heydenii species complex (Curculionidae): integrative taxonomy reveals multiple species exhibiting host specialization

A combined taxonomic, morphological, molecular and biological study revealed that the species presently named Mecinus heydenii is actually composed of five different species: M. heydenii Wencker, 1866; M. raphaelis Baviera & Caldara sp. n., M. laeviceps Tournier, 1873; M. peterharrisi Toševski & Caldara sp. n. and M. bulgaricus Angelov, 1971. These species can be distinguished from each other by a few subtle characteristics, mainly in the shape of the rostrum and body of the penis, and the colour of the integument. The first four species live on different species of Linaria plants, respectively, L. vulgaris (L.) P. Mill., L. purpurea (L.) P. Mill. L. genistifolia (L.) P. Mill. and L. dalmatica (L.) P. Mill., whereas the host plant of M. bulgaricus is still unknown. An analysis of mtCOII gene sequence data revealed high genetic divergence among these species, with uncorrected pairwise distances of 9% between M. heydenii and M. raphaelis, 11.5% between M. laeviceps, M. heydenii and M. raphaelis, while M. laeviceps and M. peterharrisi are approximately 6.3% divergent from each other. Mecinus bulgaricus exhibits even greater divergence from all these species and is more closely related to M. dorsalis Aubé, 1850. Sampled populations of M. laeviceps form three geographical subspecies: M. laeviceps laeviceps, M. laeviceps meridionalis Toševski & Jović and M. laeviceps corifoliae Toševski & Jović. These subspecies show clear genetic clustering with uncorrected mtDNA COII divergences of approximately 1.4% from each other.

Choice behaviour and performance of Cassida stigmatica on various chemotypes of Tanacetum vulgare and implications for biocontrol

Biological control is often an effective technique deployed to tackle invasive plant species by the release of highly specialized enemies. However, variation in plant chemistry due to high plasticity and/or evolved during the introduction process needs to be considered when testing the efficacy of potential biocontrol agents. The tortoise beetle, Cassida stigmatica Suffrian (Coleoptera: Chrysomelidae), is a potential biological control agent selected to control the chemically highly variable and invasive plant species Tanacetum vulgare L. (Asteraceae). This study was conducted to test whether or not plants of different chemical profiles (terpene chemotypes) are equally accepted by C. stigmatica. Therefore, female oviposition and feeding choice behaviour were examined using one pure chemotype (β‐thujone type) and three mixed chemotypes (camphor/1,8‐cineole, β‐thujone/camphor, and camphor/camphene types) in various paired‐choice combinations. Furthermore, larval survival and adult body mass were compared when beetles were reared on the four chemotypes. Overall, C. stigmatica females showed a clear preference for the pure β‐thujone chemotype over the mixed chemotypes containing camphor, and no preferences when only mixed chemotypes were offered in choice tests. However, they were able to use all offered chemotypes for oviposition and feeding. No differences in larval survival and adult body mass could be detected when reared on the various chemotypes. The study demonstrates that knowledge of the acceptance of chemically variable host species is essential when testing for applicability of biocontrol agents.