Can an a priori strategy be developed for biological control? The case of Onopordum spp. thistles in Australia

Abstract  Between 1992 and 2000, seven insect agents were released in Australia for the biological control of Onopordum spp. thistles. This paper describes the protocol used for the selection of these agents, starting with the development of a preliminary strategy, based on the ecology and population dynamics of the target weed. The strategy informed the surveys for natural enemies in the native range of Onopordum , targeting insects that attacked key transitional stages of the weed's life cycle. Ongoing studies of Onopordum populations in both Australia and Europe, plus experimental studies on the ecology, potential impact and preliminary host specificity of the agents, led to the refinement of the strategy and the selection and prioritisation of the agents. It is argued that development of an explicit strategy prior to release should be encouraged, as it forces researchers to revisit the rationale for and aims of particular biological control projects, ensuring that the process of agent selection remains focused. It also provides a tool to improve the process of agent selection, as subsequent results can be measured against the strategy and agent success or failure evaluated against the a priori expectations.     

Amplified fragment length polymorphism (AFLP) reveals introgression in weedy Onopordum thistles: hybridization and invasion

Onopordum L. (Compositae) is an extremely diverse genus of thistles, which includes several species that have become serious pasture weeds in several regions of the world. We present a comparison of the genetic diversity in invasive forms of Onopordum from Australia with several known native European species. A total of 108 polymorphic genetic markers was generated using amplified fragment length polymorphism (AFLP) fingerprinting. Non-metric multidimensional scaling (NMDS) revealed that Australia contained O. acanthium, O. illyricum and a full range of genetic intermediates between these species. Intermediates largely comprised segregating fragments diagnostic for European O. acanthium and O. illyricum with a low frequency of fragments that were diagnostic for other species never recorded in Australia. The current genetic patterns in Australia may be best explained by a combination of processes, both in the native and in the alien range. These include multiple introductions of seed, including hybrid material, and the continuous dispersal in Australia, leading to an increase in the contact among hybridizing taxa. Such processes appear to have produced more widespread hybridization and introgression in Australian Onopordum than is found in Europe.     

Comparative biology of two rosette crown-feeding flies of the genus Botanophila (Dipt., Anthomyiidae) with potential for biological control of their thistle hosts

The comparative biology of two anthomyiid flies with potential for biological control of their host thistles, Botanophila turcica on Carthamus lanatus and Botanophila spinosa on Onopordum acanthium , was studied using field surveys and collections, and by rearing collected eggs and larvae in the laboratory in southern France. The thistle hosts are significant weeds outside their native range, particularly in Australia. Both flies attack the rosette meristems of their hosts prior to flowering. Larval stages are described together with natural attack rates (21–33% of field plants) and the mortality of field-collected larvae reared in the laboratory. A successful rearing protocol for these flies is also described. The results of preliminary host-specificity tests showed that both species are highly specific, being restricted to their host genus in the tests conducted. Furthermore, Botanophila turcica could not complete development on safflower (a congener of its natural host) under natural conditions. Botanophila turcica had an adult activity period that lasted 6 months from late autumn and laid fewer eggs per host plant than B. spinosa , which was active for 2 to 3 months in spring. Only one parasitoid was observed, an endoparasitic eucoilid, Trybliographa sp., which attacked both fly species, causing 18–23% mortality.     

Chemistry of Cirsium and Carduus: a role in ecological risk assessment for biological control of weeds?

Prediction of host plant range and ecological impact of exotic phytophagous insects, such as insects for classical biological control of weeds, represents a major challenge. Recently, the flowerhead weevil (Rhinocyllus conicus Fröl.), introduced from Europe into North America to control exotic thistles (Carduus spp.), has become invasive. It feeds heavily on some, but not all species of native North American thistles (Cirsium spp.). We hypothesized that such non-target use among native plants could be better predicted by knowledge of characteristic chemical profiles of secondary compounds to supplement the results of host specificity testing. To evaluate this hypothesis, we reviewed the literature on the chemistry of Cirsium and Carduus thistles. We asked what compounds are known to be present, what is known about their biological activity, and whether such information on chemical profiles would have better predicted realized host range and ecological effects of R. conicus in North America. We found an extensive, but incomplete literature on the chemistry of true thistles. Two main patterns emerged. First, consistent chemical similarities and interesting differences occur among species of thistles. Second, variation occurs in biologically active groups of characteristic compounds, specifically flavonoids, sterols, alkaloids and phenolic acids, that are known to influence host plant acceptance, selection, and feeding by phytophagous insects. Surprisingly, sesquiterpene lactones, which are characterisitic in closely related Asteraceae, have not been extensively reported for Cirsium or Carduus. The minimal evidence on sesquiterpene lactones may reflect extraction methods vs. true absence. In summary, our review suggests further research on thistle chemistry in insect feeding is warranted. Also, since the exotic Canada thistle (Cirsium arvense) is an invasive thistle of current concern in North America, such research on mechanisms underlying host range expansion by exotic insects would be useful.     

Potential Impact of the Stem-boring Weevil Lixus cardui on the Growth and Reproductive Capacity of Onopordum Thistles

Using plants grown in large field cages, it was shown that heavy feeding by the weevil, Lixus cardui, reduced both the plant growth and reproductive capacity of Onopordum thistles. At high initial densities of adult weevils, the plant height and biomass were reduced by up to 50%. Fewer capitula were produced on Lixus -attacked plants, and these were smaller, suffered higher levels of abortion and produced ca. 80% fewer viable seeds. Attacked plants senesced 2-3 weeks earlier than unattacked plants and late-developing capitula aborted, causing a shortening of the normal flowering period. At extreme density levels, where survival of the weevils themselves was affected, damage levels were even greater. It was not possible to separate the contributions of damage by adults, which destroyed 30-40% of leaf tissue, from that by larvae, which hatched from eggs laid in the stems and mined the pith and cambium tissue. This was, however, not essential for the purpose of demonstrating the weevil's potential to contribute to biological control of the target thistles. It was considered that even a lower impact in the field would enable L. cardui to complement agents that had already been released, or were planned for release against Onopordum spp. in Australia and would reduce the competitiveness of these weeds in infested pastures.     

Host-specificity of Candidate Agents for the Biological Control of Onopordum spp. Thistles in Australia: an Assessment of Testing Procedures

This paper describes a series of host-specificity tests carried out to evaluate the safety of eight insects for release in Australia as biological control agents of Onopordum spp. thistles; Larinus latus L., Lixus cardui Ol. and Trichosirocalus briesei Alonso-Zarazaga & Sanchez-Ruiz (Coleoptera: Curculionidae), Tephritis postica Loew and Urophora terebrans Loew (Diptera: Tephritidae), Botanophila spinosa Rondani (Diptera: Anthomyiidae), Tettigometra sulphurea Mulsant & Rey. (Hemiptera: Fulgoroidae) and Eublemma amoena (Hbn.) (Lepidoptera: Noctuidae). Preliminary tests were first carried out against key test plants in the country of origin to determine whether it was worth proceeding with formal testing under quarantine conditions in Australia. In the formal testing procedure, the test plant list was based solely on phylogenetic relationships. The life-stage of each insect tested was selected with respect to its biology and behaviour, e.g. only oviposition was tested for insect species with immobile endophagous larvae, while larval host utilization was tested for those with mobile larval stages. The initial laboratory experiments for each species used were caged no-choice tests, as confinement without choice can elicit extreme behavioural responses, making negative results extremely robust. Where positive results occurred, insects where re-exposed to the plant, but this time given a choice between the target and non-target plants. The results of these tests, plus data from open-field preliminary tests in the country of origin, helped interpret results and determine the risk posed by each candidate biological control agent to non-target plant species. Following submission, all eight candidates were approved for release by the Australian plant biosecurity and conservation authorities. Based on a comparison of the eight species tested, it is recommended that host-specificity testing be kept flexible and pragmatic, rather than moving towards a formulaic risk assessment procedure. Moreover, given the improved state of knowledge of plant phylogenies and the evolution of host utilization, it is time to base testing procedures purely on phylogenetic grounds, without the need to include less related test species solely because of economic or conservation reasons.     

The taxonomic status of Larinus cynarae F. and Larinus latus Herbst (coleoptera: Curculionidae), and its implications for the biological control of Onopordum (asteraceae: Cardueae) in Australia

Analyses were made of the allozyme frequencies of sympatric populations of the capitulum weevils L. latus Herbst and Larinus cyarae F. in Greece. It was found that the two taxa are genetically isolated and that they can therefore be considered as separate species. This complemented ecological data showing separation of the L. latus and L. cynarae populations by their choice of Onopordum illyricum L. and Cynara cardunculus, respectively, as hosts. Onopordum spp. are serious weed problems in Australia while Cynara contains several crop plants. L. latus is a potential biological control agent of Onopordum spp., but its use had been compromised by the uncertainty of the taxonomic distinctness from L. cynarae, biotypes of which are pests of artichoke Cynara scolymus L. Clarification of its taxonomic status lifts these reservations and clears the way for the introduction of L. latus into Australia for the biological control of Onopordum thistles.     

Faunal make-up, host range and infestation rate of weevils and tephritid flies associated with flower heads of the thistle Cirsium (Cardueae: Astaraceae) in Japan

From 1988 to 1998, we collected flower heads of 39 thistle taxa (35 taxa of Cirsium, one species each of Breea, Synurus, Saussurea and Arctium; Cardueae; Astaraceae) in Japan, mainly from Hokuriku and other parts of central Honshu, and kept them in the laboratory to breed weevils and tephritid flies, the core fauna. We report the faunal make‐up, host plants, geographic distribution and the attack levels of the insects. Results indicated that (i) three Larinus species (Curculionidae) and three species of tephritid flies (Tephritis, Urophora and Xyphosia) comprised the core fauna; (ii) two insect species belonging to the same taxonomic group (either Curculionidae (Larinus) or Tephritidae) tended to use different host plant species; (iii) two sympatric Larinus species (L. latissimus and L. meleagris) segregated the host plants seasonally in central Honshu (Cirsium blooming in spring and autumn, respectively); and (iv) two tephritid fly species, Xyphosia punctigera and Urophora sachalinensis, segregated geographically (the former on the Japan Sea side and the latter on the Pacific Ocean side). In comparison with their European counterparts, the weevils and tephritids of the Japanese Cirsium are characterized by a lower species richness and a lower degree of specialization in usage of the thistle flower heads, with gall‐formers being distinctly under‐represented, and callus tissue‐feeders being absent. This reflects the fact that Japanese thistles are so closely related that hybridization frequently occurs, and also that the thistles have had a short history of interaction with the insects since the thistles’ arrival in Japan.     

Cryptic local adaptation of the herbivorous ladybird beetle Henosepilachna pustulosa in the ability to use the thistle Cirsium boreale

Local adaptation to different host plants is important in the diversification of phytophagous insects. Thus far, much evidence of the local adaptation of insects with respect to host use at the physiological level has been gathered from systems involving less mobile insects and/or divergent hosts such as plants belonging to different families or genera. On the other hand, the prevalence of such local adaptation of insects with moderate or high dispersal ability to the intraspecific variation of herbaceous hosts is largely unknown. In the present study, we examined the occurrence and degree of local adaptation of the herbivorous ladybird beetle Henosepilachna pustulosa (Kôno) (Coleoptera: Coccinellidae) to its primary host, the thistle Cirsium boreale Kitam. (Asteraceae), through reciprocal laboratory experiments using beetles and thistles from three locations with a range of approximately 200 km. Concerning the larval developmental ability, obvious patterns of local adaptation to the thistles from respective natal locations were detected, at least in some combinations of beetle populations. Similar tendencies were detected concerning adult feeding acceptance, although the statistical support was somewhat obscure. Overall, our results indicate that the degree of local adaptation of insect species with moderate dispersal ability to conspecific herbaceous hosts is occasionally as strong as that involving less mobile insects and/or heterospecific hosts, indicating the potential of such cryptic local adaptation to promote ecological/genetic differentiation of phytophagous insect populations.     

Herbivorous arthropods on bracken (Pteridium aquilinum (L.) Khun) in Australia compared with elsewhere

Abstract The herbivorous arthropod fauna of bracken fern (Pteridium aquilinum (L.) Khun), at a site near Sydney, Australia, is described and compared with previously reported bracken faunas in other geographic regions. Monthly sampling over 18 months found 17 species of herbivorous arthropods (15 insect and two mite species) from five orders. At the ordinal level, the mixture differed substantially from the bracken faunas of sites in Britain and Papua New Guinea. Notable was the presence of Thysanoptera and Acari, and the absence of Coleoptera and Hymenoptera. The mixtures of orders/families represented in the site bracken faunas in Britain, and less so in Australia, resembled those in the pool of herbivorous arthropods in those regions. Further, the mixture of orders on bracken was more similar to the mixture of orders on other ferns than to the mixture of orders among herbivorous insects on all plants; such similarity was not evident at family level. Compared with sites in other regions, the Sydney site had an abundance of pinna-sucking species and a dearth of mining species. Differences between regions in feeding niches most occupied tended to correspond with the differences in orders represented. Not all features of the fauna of bracken near Sydney reflected differences in the general herbivorous arthropod fauna of Australia compared with other regions, or differences between the herbivore faunas of ferns and seed plants. Its composition must be attributed in part to stochastic aspects of the speciation of herbivorous arthropods onto host plants.     

Phytophagous insect assemblages and the regional species pool: patterns and asymmetries

We describe three models predicting relationships between: (a) the taxonomic composition of the regional species pool of phytophagous insects and the composition of the phytophagous insect fauna on a host taxon; and (b) the faunal composition of two host taxa. The predictions of these models were compared with empirical data representing the regional pool of phytophages in Central Europe and the faunas of two plant taxa: the cabbage plants (Brassicaceae) and the thistles (Asteraceae: Cardueae). Three important findings emerge at a general level. (1) Different taxonomic levels of insects (orders, families, genera) of the regional pool and on the investigated host taxa are well correlated in terms of species richness, but there is no consistent trend in the variance explained by this correlation across taxonomic levels. (2) The model considering evolutionary interactions and speciation processes is consistent with patterns found in the empirical data. (3) Asymmetries in sampled species numbers of insect families on both host taxa may be accounted for by reference to the biology of these insects. We conclude that the faunas of single host taxa can provide the basis for extrapolating to the regional pool, at least at high taxonomic levels.     

Leaf miners: The hidden herbivores

Leaf mining is a form of endophagous herbivory in which insect larvae live and feed within leaf tissue. In this review we discuss aspects of leaf miner ecology, and the current evidence for three hypotheses relating to the evolution of this feeding guild. We also present a summary of the literature coverage relating to these herbivores, which have been relatively poorly studied compared with insects that feed externally such as sap suckers and leaf chewers. The majority of published studies concern leaf miners from the northern hemisphere, with a general focus on those species considered to be agricultural, forestry or horticultural pests. In a more detailed case study, we examine aspects of leaf miner ecology of Australian species. At least 114 species have been recorded as leaf miners in Australia in four orders: Coleoptera, Lepidoptera, Diptera and Hymenoptera. Lepidoptera and Diptera are the most speciose orders of Australian leaf miners; Hymenoptera are represented by a single endemic genus and half of all coleopteran miners are species introduced for biological control. Both the known number of leaf‐mining species in Australia and the known number of hosts have increased in recent years following new targeted surveys. Leaf miners in Australia occur in many habitats and feed on a wide variety of host plants in at least 60 families although most individual species are monophagous. Although much of the research on leaf miners in Australia has focused on species that are commercially important pests or biological control agents, studies on fundamental aspects of leaf miner ecology are increasing. We identify a number of research questions aimed at better understanding the ecology of leaf miners in Australia and elsewhere.     

Ontogenetic diet shifts in Roeboides affinis with morphological comparisons

Fishes of the characid genus Roeboides are documented as lepidophagous, but the degree of specialization on scales varies between species. This study examines ontogenetic changes in morphology and diet of Roeboides dayi and Roeboides affinis from Venezuela. Juveniles of both species feed on microcrustacea and insects during the wet season, when availability of these resources is greatest. As aquatic habitat decreases during the dry season, insect and microcrustacea availability decreases and fish densities increase. As the fish densities increase, scale consumption becomes more profitable. Interspecific differences in the degree of specialization of the teeth and jaws was associated with the degree of scale feeding. Adult R. dayi consumed scales in approximately equal proportions to insects. Adult R. affinis consumes approximately 100% scales, and has more specialized head morphology than R. dayi.     

Phylogenetic relationships in the Neotropical bruchid genus Acanthoscelides (Bruchinae, Bruchidae, Coleoptera)

Adaptation to host-plant defences through key innovations is a driving force of evolution in phytophagous insects. Species of the neotropical bruchid genus Acanthoscelides Schilsky are known to be associated with specific host plants. The speciation processes involved in such specialization pattern that have produced these specific associations may reflect radiations linked to particular kinds of host plants. By studying host-plant associations in closely related bruchid species, we have shown that adaptation to a particular host-plant (e.g. with a certain type of secondary compounds) could generally lead to a radiation of bruchid species at the level of terminal branches. However, in some cases of recent host shifts, there is no congruence between genetic proximity of bruchid species, and taxonomic similarity of host plants. At deeper branches in the phylogeny, vicariance or long-distance colonization events seem to be responsible for genetic divergence between well-marked clades rather than adaptation to host plants. Our study also suggests that the few species of Acanthoscelides described from the Old World, as well as Neotropical species feeding on Mimosoideae, are misclassified, and are more closely related to the sister genus Bruchidius .     

Pattern and process in the evolution of insect-plant associations: Yponomeuta as an example

Phylogenetic studies are increasing our understanding of the evolution of associations between phytophagous insects and their host plants. Sequential evolution, i.e. the shift of insect herbivores onto pre-existing plant species, appears to be much more common than coevolution, where reciprocal selection between interacting insects and plants is thought to induce chemical diversification and resistance in plants and food specialization in insects.Extreme host specificity is common in phytophagous insects and future studies are likely to reveal even more specialization. Hypotheses that assume that food specialists have selective advantages over generalists do not seem to provide a general explanation for the ubiquity of specialist insect herbivores. Specialists are probably committed to remain so, because they have little evolutionary opportunity to reverse the process due to genetically determined constraints on the evolution of their physiology or nervous system. The same constraints might result in phylogenetic conservatism, i.e. the frequent association of related insect herbivores with related plants. Current phylogenetic evidence, however, indicates that there is no intrinsic direction to the evolution of specialization.Historical aspects of insect-host plant associations will be illustrated with the small ermine moth genus Yponomeuta. Small ermine moths show an ancestral host association with the family Celastraceae. The genus seems to be committed to specialization per se rather than to a particular group of plants. Whatever host shift they have made in their evolutionary past (onto Rosaceae, Crassulaceae, and Salicaceae), they remain monophagous. The oligophagous Y. padellus is the only exception. This species might comprise a mosaic of genetically divergent host-associated populations.     

Local adaptation and ecological genetics of host-plant specialization in a leaf beetle

The tendency of insect species to evolve specialization to one or a few plant species is probably a major reason for the remarkable diversity of herbivorous insects. The suggested explanations for this general trend toward specialization include a range of evolutionary mechanisms, whose relative importance is debated. Here we address two potentially important mechanisms: (i) how variation in the geographic distribution of host use may lead to the evolution of local adaptation and specialization; (ii) how selection for specialization may lead to the evolution of trade‐offs in performance between different hosts. We performed a quantitative genetic experiment of larval performance in three different populations of the alpine leaf beetle Oreina elongata reared on two of its main host plants. Due to differences in host availability, each population represents a distinctly different selective regime in terms of host use including selection for specialization on one or the other host as well as selection for utilizing both hosts during the larval stage. The results suggest that selection for specialization has lead to some degree of local adaptations in host use: both single‐host population had higher larval growth rate on their respective native host plant genus, while there was no difference between plant treatments in the two‐host population. However, differences between host plant treatments within populations were generally small and the degree of local adaptation in performance traits seems to be relatively limited. Genetic correlations in performance traits between the hosts ranged from zero in the two‐host population to significantly positive in the single‐host populations. This suggests that selection for specialization in single host populations typically also increased performance on the alternative host that is not naturally encountered. Moreover, the lack of a positive genetic correlation in the two host‐population give support for the hypothesis that performance trade‐offs between two host plants may typically evolve when a population have adapted to both these plants. We conclude that although there is selection for specialization in larval performance traits it seems as if the genetic architecture of these traits have limited the divergence between populations in relative performance on the two hosts.     

Host utilization of field-caged native and introduced thistle species by Rhinocyllus conicus

Rhinocyllus conicus Fr?elich was introduced from Europe into North America as a biological control agent of the exotic weed Carduus nutans L. Concern exists over the feeding of this weevil on at least 25 species of native Cirsium thistles. Beginning in 2008, cage studies isolating adults of R. conicus on buds and flower heads of all eight thistle species (native and introduced) recorded from Tennessee were conducted to test if R. conicus could use these species for reproduction and what impacts larval feeding of R. conicus may have on seed production. Larvae of R. conicus completed development in heads of the native species C. carolinianum (Walter) Fernald and Schubert. and C. horridulum Michaux, and significant reductions in seed numbers of both species occurred during 2008. Rhinocyllus conicus oviposited on both C. carolinianum and C. horridulum at significantly greater levels than the introduced species C. arvense (L.) Scopoli and C. vulgare (Savi) Tenore. Infested heads of C. carolinianum contained numbers of R. conicus per centimeter of plant head width similar to Ca. nutans in 2008, and both native species contained numbers of R. conicus per centimeter of plant head width similar to C. arvense and C. vulgare in 2009. Body length was similar between R. conicus reared on native thistles and its target host Ca. nutans. This report is the first documentation of R. conicus feeding and reproducing on C. carolinianum and C. horridulum. Although R. conicus has been observed only on introduced thistles in naturally occurring populations in this region, the utilization of C. carolinianum and C. horridulum as host species in controlled conditions warrants continued monitoring of field populations and further investigation into factors that may influence nontarget feeding in the future.     

Exotic thistles increase native ant abundance through the maintenance of enhanced aphid populations

Exotic species change the structure and composition of invaded communities in multiple ways, but the sign of their impact on native species is still controversial. We evaluated the effects of the thistles Carduus thoermeri and Onopordum acanthium—two of the most abundant exotic plant species in disturbed areas of the Patagonian steppe—on the native tending ant assemblage. Exotic thistles showed an increased number of plants with aphids and had greater aphid density than native plants. Since native tending ants were present only in plants with aphids, their abundance was higher in infested thistles than in native plants. Path analyses confirmed that ant activity depended more on aphid density than on thistle traits. Our results suggest that the presence of exotic thistles in disturbed areas of NW Patagonia indirectly benefit the native ant assemblage through the maintenance of an increased aphid population. This illustrates how the impact of exotic on native species can depend on the ecological context.     

Host selection patterns in insects breeding in bracket fungi

Abstract.  1. Fungivorous insects are generally viewed as polyphagous, largely because most fungal fruiting bodies constitute an unpredictable resource. To examine the validity of this hypothesis, and degree of phylogenetic relatedness between the preferred hosts of the insects, host selection in the insect fauna of bracket fungi was studied, using data obtained both from the field and the literature.
2. More than half (53%) of the insect species breeding in them appeared to be monophagous.
3. Modern phylogenies explained the host selection patterns better than older classifications, since non-monophagous species of beetles frequently used hosts that are closely related to each other.
4. The hypothesis that polyphagous species use more heavily decayed fruiting bodies than monophagous species was verified for insects breeding in Fomes fomentarius . The results indicate that the chemical composition of the fungi influences host selection.
5. It is suggested that fruiting bodies of bracket fungi differ from most other fungi in that their occurrence is more predictable. Therefore, the primary colonising fungivores generally attack only one host species, or a few hosts that are closely phylogenetically related. Polyphagous species generally colonise fruiting bodies after they have reached a certain stage of decay, thus escaping their chemical defence.