Candidate Biological Control Agent Research Articles

Potential biological control agents of Geosmithia morbida restrict fungal pathogen growth via mycoparasitism and antibiosis

Thousand cankers disease of Juglans (walnut) and Pterocarya (wingnut) spp. (Fagales: Juglandaceae) is caused by the fungal pathogen Geosmithia morbida Kolarík, Freeland, Utley, and Tisserat (Hypocreales: Bionectriaceae) and bark beetle pest/vector, Pityophthorus juglandis Blackman (Coleoptera: Curculionidae). To further the development of biological management strategies for thousand cankers disease, we assessed the ability of 14 endophytic Trichoderma (Hypocreales: Hypocreaceae) isolates and the commercially available isolate T. afroharzianum strain KRL-AG2 to inhibit the in vitro growth of three different G. morbida isolates via mycoparasitism and antibiosis. To identify factors that may affect field success of candidate biological control agents, we quantified the growth responses of Trichoderma spp. and the commercially available entomopathogenic fungus, Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) strain GHA, to the plant secondary metabolite and antimicrobial compound, juglone in vitro. A total of 12 Trichoderma isolates (from six different Trichoderma species) demonstrated antagonistic activity towards G. morbida in dual-plate assays. Juglone consistently reduced the growth of B. bassiana strain GHA and 14 of the 15 screened Trichoderma isolates in vitro. Additionally, one metabolite-producing Trichoderma isolate, TN4-47, completely inhibited the growth of all three G. morbida isolates across all tested metabolite concentrations and had comparatively greater tolerance to juglone compared to other Trichoderma isolates. Future lines of research should focus on characterizing the active antagonistic compound present in the metabolite filtrates, determine the mode of action of the active component(s), and elucidate how abiotic and biotic factors may influence the growth, persistence, and antagonistic activity of candidate biological control agents in planta.

Insights to the host range of the flea beetle Disonycha argentinensis Jacoby, 1992 (Coleoptera: Chrysomelidae), a candidate biological control agent of alligator weed in the U.S.A.

Insights to the host range of the flea beetle Disonycha argentinensis Jacoby, 1992 (Coleoptera: Chrysomelidae), a candidate biological control agent of alligator weed in the U.S.A.

Genetic Diversity of Ralstonia solanacearum Causing Tobacco Bacterial Wilt in Fujian Province and Identification of Biocontrol Streptomyces sp.

Tobacco bacterial wilt is a highly destructive soilborne disease caused by the Ralstonia solanacearum species complex, exhibiting a significant risk to global flue-cured tobacco cultivation and resulting in substantial economic loss. In this study, 77 isolates were collected from three prominent flue-cured tobacco cultivation areas in Fujian, China (Nanping, Sanming, and Longyan), in 2021 and 2022. The isolated strains were classified through phylotype-specific multiplex polymerase chain reaction (Pmx-PCR) and physiological tests. The analysis showed that all the strains were associated with phylotype I, race 1, and biovar III. Subsequent phylogenetic analysis using partial egl gene sequences classified the 77 isolates into 5 distinct sequevars: 13, 15, 16, 17, and 34. Notably, a remarkable predominance of sequevar 15 was observed in Fujian Province, while sequevar 16 was first reported on tobacco in China, which was identified in other plants, expanding the understanding of its host range and distribution in the country. In addition, a Streptomyces strain extracted from the rhizosphere soil of tobacco was found to inhibit the growth of multiple sequevars of tobacco R. solanacearum, indicating its broad-spectrum antagonistic properties. Furthermore, pot experiments showed that the strain St35 effectively controlled tobacco bacterial wilt. The isolate St35 was conclusively identified as Streptomyces gancidicus according to the morphological and genetic features. In summary, the present study demonstrated the genetic diversity and distribution of tobacco R. solanacearum strains in the Fujian province of China, as well as the identification of a candidate biological control agent for the management of tobacco bacterial wilt.

In-vitro endophytic bacteria activity against Ralstonia syzygii subsp. indonesiensis, the causal agent of bacterial wilt disease in tomato (Solanum lycopersicum) to achieve sustainable agriculture

The bacterial wilt disease transmitted through soil, identified as Ralstonia syzygii subsp. indonesiensis, is a substantial vascular disease and major constraint in various Solanaceae plants. The use endophytic bacteria-as Arthrobacter sp. and Bacillus thuringiensis-is one approach to control this pathogen. In vitro test using the disc paper method was conducted to evaluate the compatibility of Arthrobacter sp. and B. thuringiensis, along with their inhibitory capabilities through secondary metabolite production, in suppressing the growth of R. syzygii subsp. indonesiensis. The results showed incompatibility among isolates of endophytic bacterial, consequently, both endophytic bacteria were capable suppressing the growth of the R. syzygii subsp. indonesiensis significantly. Arthrobacter sp. formed a larger inhibition zone compared to B. thuringiensis, and both endophytic bacteria can be candidate biological control agents to reduce pesticide use to control R. syzygii subsp. indonesiensis, which therefore achieving more environmentally agriculture sustainability.

A small step to discover candidate biological control agents from preexisting bioresources by using novel nonribosomal peptide synthetases hidden in activated sludge metagenomes.

Biological control agents (BCAs), beneficial organisms that reduce the incidence or severity of plant disease, have been expected to be alternatives to replace chemical pesticides worldwide. To date, BCAs have been screened by culture-dependent methods from various environments. However, previously unknown BCA candidates may be buried and overlooked because this approach preferentially selects only easy-to-culture microbial lineages. To overcome this limitation, as a small-scale test case, we attempted to explore novel BCA candidates by employing the shotgun metagenomic information of the activated sludge (AS) microbiome, which is thought to contain unutilized biological resources. We first performed genome-resolved metagenomics for AS taken from a municipal sewage treatment plant and obtained 97 nonribosomal peptide synthetase (NRPS)/polyketide synthase (PKS)-related gene sequences from 43 metagenomic assembled bins, most of which were assigned to the phyla Proteobacteria and Myxococcota. Furthermore, these NRPS/PKS-related genes are predicted to be novel because they were genetically dissimilar to known NRPS/PKS gene clusters. Of these, the condensation domain of the syringomycin-related NRPS gene cluster was detected in Rhodoferax- and Rhodocyclaceae-related bins, and its homolog was found in previously reported AS metagenomes as well as the genomes of three strains available from the microbial culture collections, implying their potential BCA ability. Then, we tested the antimicrobial activity of these strains against phytopathogenic fungi to investigate the potential ability of BCA by in vitro cultivation and successfully confirmed the actual antifungal activity of three strains harboring a possibly novel NRPS gene cluster. Our findings provide a possible strategy for discovering novel BCAs buried in the environment using genome-resolved metagenomics.

Calosoma (Coleoptera: Carabidae) biological control introductions to Hawaiʻi: Taxonomic ignorance and confusion lead to an unexpected result

From 1897 until 1957, beetles of the genus Calosoma L. (Coleoptera: Carabidae) were repeatedly introduced to the Hawaiian Islands as predators of nutgrass armyworm, Spodoptera exempta (Walker), armyworm, Mythimna unipuncta (Haworth), and lawn armyworm, Spodoptera mauritia (Boisduval). Through examination of institutional voucher specimens, archival field notes, and the literature, we document failed biological control introductions of eight flight-capable Calosoma species: C. calidum (F.), C. frigidum Kirby, C. marginale Casey, C. peregrinator Guérin-Méneville, C. prominens LeConte, C. protractum LeConte, C. semilaeve LeConte, and C. senegalense Dejean. In contrast, the vestigially-winged C. anthracinum Dejean, a rarely collected Mexican species never intentionally introduced, is the only Calosoma currently resident in Hawaiʻi. Calosoma anthracinum was misidentified initially as C. semilaeve, and thereafter misidentified as Calosoma blaptoides tehuanacum Lapouge. Both C. anthracinum and C. protractum were collected sympatrically at Cuernavaca, Morelos, Mexico during Hawaiʻi-based explorations, and we hypothesize that they were introduced together as a species mixture in a 1923 release made at Waikiʻi, Parker Ranch, Hawaiʻi Island. Calosoma anthracinum was little known to taxonomists at that time due to its limited native distribution, rarity in institutional collections, and absence from taxonomic keys, leading to repeated misidentifications. Calosoma peregrinator and C. prominens, two abundant, sympatric species from the Sonoran and Chihuahuan Deserts, were also taxonomically confused over this period, resulting in mixed-species Hawaiian introductions. Three species that failed to establish upon introduction—C. marginale, C. peregrinator and C. semilaeve—have been intercepted as singletons or pairs of individuals decades later, without apparent establishment. Calosoma beetles’ ecological requirements, dispersal abilities, introduction histories, and native ranges and associated climate are compared among species, the only unique characteristic of the successful colonist being flightlessness. The history of Hawaiian Calosoma introductions illustrates the need for robust and accessible classifications based on accurately determined specimens as well as comprehensive understanding of the biological attributes and habitat requirements of candidate biological control agents.

Guidelines and framework to assess the feasibility of starting pre-emptive risk assessment of classical biological control agents

Non-native invasive arthropod species threaten biodiversity and food security worldwide, resulting in substantial economic, environmental, social and cultural costs. Classical biological control (CBC) is regarded as a cost-effective component of integrated pest management programmes to manage invasive arthropod pests sustainably. However, CBC programmes are traditionally conducted once a pest has established in a new environment, and invariably all research needed to achieve approval to release a biological control agent can take several years. During that time, adverse impacts of the pest accelerate. A pre-emptive biocontrol approach will provide the opportunity to develop CBC for invasive pests before they arrive in the country at risk of introduction and therefore enhance preparedness. A critical aspect of this approach is that risk assessment is carried out in advance of the arrival of the pest. Implementing pre-emptive biocontrol risk assessment means that natural enemies can be selected, screened in containment or abroad and potentially pre-approved prior to a pest establishing in the country at risk, thus improving CBC effectiveness. However, such an approach may not always be feasible. This contribution defines the fundamental prerequisites, principles, and objectives of pre-emptive biocontrol risk assessment. A set of guidelines and a decision framework were developed, which can be used to assess the feasibility of conducting a pre-emptive risk assessment for candidate biological control agents against high-risk arthropod pests.

Exploring Wild and Local Fruits as Sources of Promising Biocontrol Agents against Alternaria spp. in Apples

Biological control agents (BCAs) are a promising option for managing postharvest diseases. Their environmentally friendly nature makes them valuable for sustainable and eco-friendly postharvest disease management. This study evaluated the antagonistic potential of epiphytic yeasts isolated from a local apple genotype known as “Niğde Elması” and a range of wild fruits: rosehip, hawthorn, and wild pear. There were 375 yeast isolates obtained and screened in vitro and in vivo. Initially selected were 32 isolates able to inhibit the growth of Alternaria alternata mycelia in the in vitro experiments and identified using molecular methods as candidate BCAs. These isolates were identified as Aureobasidium pullulans using sequence analysis of the internal transcribed spacer (ITS) and the translation elongation factor EF-1α gene (EF1α). Based on two rounds of in vivo screening, four isolates were subsequently selected for their capability to control the infection of apple fruits under high inoculum pressure. Finally, their volatile and non-volatile antimicrobial activity was tested against Alternaria spp. These findings showed how wild fruits and a local apple genotype could represent a promising source for new BCA isolation. However, further studies are needed to reveal the mechanisms of action of these putative BCAs for application during the postharvest processing and storage of apples.

Ectoparasitoid Dinarmus basalis causes greater offspring loss to the winged morph of Callosobruchus maculatus

The pest cowpea weevil Callosobruchus maculatus (F.) (Coleoptera: Chrysomelidae) exhibits polymorphism with flight and flightless forms that differ in morphology and life-history. Flight forms are generally dispersers with lower fecundity that increase in frequency when population density and intraspecific competition are both high. Callosobruchus maculatus larvae can cause important damage to stored grains and the solitary ectoparasitoid Dinarmus basalis (Rondani) (Hymenoptera: Pteromalidae) is considered to be a good candidate biological control agent. However, whether morphs differ in the likelihood of their offspring surviving attack is unknown. Understanding this has implications for the pest status of the cowpea weevil following biological control. We found that attack by D. basalis lowered the number of emerging offspring produced by both morphs. The ectoparasitoid was most effective at reducing survival of larvae produced by flight morph parents, suggesting that there may be a further cost of dispersal for C. maculatus. Parental morph type did not influence D. basalis behavior or development when foraging, so the biological mechanism resulting in this difference remains unclear. Nevertheless, our study shows that D. basalis affects the offspring of both weevil morphs, as required for an effective biocontrol agent. These results contribute to our understanding of how intraspecific variation, including polymorphism, influences species interactions among biological control agents and their target insect pests.

Synergism of a mixed diet of Myzus persicae and egg of Ephestia kuehniella on fitness of the predator Nabis stenoferus

Nabis stenoferus is a zoophytophagous predator that lives in grasslands around agricultural fields. It is a candidate biological control agent for use via augmentation or conservation. To find a suitable food source for mass-rearing and to better understand this predator’s biology, we compared the life history characteristics of N. stenoferus under the three different diets: aphids only (Myzus persicae), moth eggs only (Ephestia kuehniella), or a mixed diet of aphids and moth eggs. Interestingly, when only aphids were supplied, N. stenoferus developed to the adult stage but lacked normal levels of fecundity. There was a significant synergism of the mixed diet on N. stenoferus fitness in both the immature and adult stages, i.e., a 13% reduction in the nymphal developmental period and an 87.3-fold increase in fecundity, compared to aphid-only diet. Furthermore, the intrinsic rate of increase was significantly higher for the mixed diet (0.139) than either aphids only (0.022) or moth eggs only (0.097). These results show that M. persicae alone is not a complete diet for the mass-rearing of N. stenoferus, whereas this aphid can be a supplementary food when combined with E. kuehniella eggs. Implications and applications of these findings for biological control are discussed.

Partial refuges from biological control due to intraspecific variation in protective host traits.

Predicting how much of a host or prey population may be attacked by their natural enemies is fundamental to several subfields of applied ecology, particularly biological control of pest organisms. Hosts or prey can occupy refuges that prevent them from being killed by natural enemies, but habitat or ecological refuges are challenging or impossible to predict in a laboratory setting-which is often where efficacy and specificity testing of candidate biological control agents is done. Here we explore how intraspecific variation in continuous traits of individuals or groups that confer some protection from natural enemy attack-even after the natural enemy has encountered the prey-could provide partial refuges. The size of these trait-based refuges (i.e., the proportion of prey that survive natural enemy encounters due to protective traits) should depend on the relationship between trait values and host/prey susceptibility to natural enemy attack and on how common different trait values are within a host/prey population. These can be readily estimated in laboratory testing of natural enemy impact on target or nontarget prey or hosts as long as sufficient host material is available. We provide a general framework for how intraspecific variation in protective host traits could be integrated into biological control research, specifically with reference to nontarget testing as part of classical biological control programs. As a case study, we exposed different host clutch sizes of target (pest) and nontarget (native species) stink bug (Hemiptera: Pentatomidae) species to a well-studied exotic biocontrol agent, the egg parasitoid Trissolcus japonicus (Hymenoptera: Scelionidae). We predicted that the smallest and largest clutches would occupy trait-based refuges from parasitism. Although we observed several behavioral and reproductive responses to variation in host egg mass size by T. japonicus, they did not translate to increases in host survival large enough to change the conclusions of nontarget testing. We encourage researchers to investigate intraspecific variation in a wider variety of protective host and prey traits and their consequences for refuge size.

Genetic relationships among laboratory lines of the egg parasitoid Trissolcus japonicus from native and adventive populations

Candidate biological control agents of invasive insect pests are increasingly being found in new geographic regions as a result of unintentional introductions. However, testing the degree of genetic differentiation among adventive and native-range populations of these agents is rarely done. We used reduced-representation sequencing of genomic DNA to investigate the relationships among laboratory lines of Trissolcus japonicus (Ashmead) (Hymenoptera, Scelionidae), an egg parasitoid and biological control agent of the brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera, Pentatomidae). We compared sequences from multiple adventive populations in North America (Canada, USA) and Europe (Switzerland) with populations sourced from part of its native range in China. We found considerably more genetic variation among lines sourced from adventive populations than among those within native populations. In the Pacific Northwest of North America (British Columbia, Canada and Washington State, USA), we found preliminary evidence of three distinct genetic clusters, two of which were highly dissimilar from all other lines we genotyped. In contrast, we found that other adventive lines with close geographic proximity (two from Ontario, Canada, three from Switzerland) had limited genetic variation. These findings provide a basis for testing biological differences among lines that will inform their use as biological control agents, and provide evidence to support a hypothesis of several independent introductions of T. japonicus in western North America from different source areas.

Factors Affecting Progeny Production and Sex Ratio of Gryon aetherium (Hymenoptera: Scelionidae), a Candidate Biological Control Agent for Bagrada hilaris (Hemiptera: Pentatomidae).

Manipulating the factors that influence progeny production and sex ratio in parasitoids can help maximize the production of quarantine bioassays and/or mass releases. In a series of experiments, we studied the effects of several factors on offspring production and sex ratio in the parasitoid Gryon aetherium (Hymenoptera: Scelionidae), a candidate biological control agent for Bagrada hilaris (Hemiptera: Pentatomidae). Progeny production was influenced by maternal age and dropped when females were 24 or 28 days old and decreased on the second day of exposure. Overall, the offspring sex ratio was highly variable in G. aetherium and was affected by the duration of exposure, with higher proportions of females emerging after one day of exposure than after two days, but was not affected by female density, female age/host deprivation, or temperature during oviposition. Progeny production was affected by the temperature during oviposition and was highest at 26.6 °C. The results indicate that production of G. aetherium can be maximized at one day of exposure, using females that are less than 24 d old, and at temperatures of around 26 °C.

Pathogenicity of La Jolla Virus in Drosophila suzukii following Oral Administration.

Drosophila suzukii (Ds) is an invasive pest insect that causes severe and widespread damage to soft fruit crops. Chemical control based on topical insecticides is inefficient and harmful to consumers and the environment, prompting interest in the development of biological control measures such as insect viruses with narrow host specificity. We previously described a strain of La Jolla virus (LJV) found in moribund Ds specimens in Germany. We demonstrated a pathogenic effect following the intrathoracic injection of LJV into adult Ds flies. However, the development of an effective biocontrol product based on LJV would require the characterization of (1) virulence following oral delivery, particularly in larvae, and (2) stability under different pH and temperature conditions reflecting realistic exposure scenarios. Here we describe the pathogenicity of LJV following oral delivery to Ds adults and larvae. The oral infection of Ds adults with LJV reduced survival in a concentration-dependent manner, whereas the oral infection of Ds larvae caused the arrest of development during pupation. LJV remained stable and infectious following exposure to a broad pH range and different temperatures. We, therefore, demonstrated that LJV is promising as a candidate biological control agent against Ds.

Native range efficacy assessment of Calophya terebinthifolii , a candidate biological control agent of Schinus terebinthifolia in Florida, USA

Background Aroeira or Brazilian peppertree, Schinus terebinthifolia Raddi (Anacardiaceae: Rhoeae), is one of the worst invasive plants in California, Florida, and Hawaii, USA because of its wide distribution, adaptation to various habitats, toxicity, and demonstrated negative impacts on biodiversity. Chemical and mechanical methods provide only temporary control, are expensive, may damage native plants, and often create disturbance conducive to reinvasion by Brazilian peppertree and other invasive weeds. The leaflet galling psyllid Calophya terebinthifolii Burckhardt & Basset (Calophyidae: Calophyinae) is [...]

Life Histories and Host-Range Evaluation of Two Chrysomelid Beetles (Zygogramma spp.) Released against Tithonia rotundifolia in South Africa.

Simple SummaryTithonia rotundifolia (Mill.) S.F. Blake (Asteraceae) is among the three Tithonia species from Mexico that are invasive in many countries, including South Africa. To curb the invasiveness and negative impact of T. rotundifolia in South Africa, two chrysomelid beetles, Zygogramma signatipennis (Stål) and Zygogramma piceicollis (Stål), from Mexico were assessed to determine their suitability for release against this invader. Biological attributes such as a short pre-oviposition period, short egg incubation period, short lifecycle and long longevity suggest that the two beetle species could successfully establish in their introduced range. Feeding, oviposition and development of both beetle species were confined within the tribe Heliantheae, but showed a very strong preference for the invasive T. rotundifolia. The only non-target species that supported development to adulthood was the exotic weed Tithonia diversifolia (Hemsl.) A. Gray, itself a target for biocontrol. Although some sunflower (Helianthus annuus L.) cultivars were partially utilized during host-specificity tests, none supported complete development. Since their potential threats to H. annuus cultivars are minimal, both Zygogramma species were cleared for release in South Africa in 2014. Tithonia rotundifolia (Mill.) S.F. Blake (Asteraceae) is among the three Tithonia species from Mexico that are invasive in South Africa. To curb its invasiveness and negative impact in South Africa, two chrysomelid beetles, Zygogramma signatipennis (Stål) and Zygogramma piceicollis (Stål), from Mexico were investigated as candidate biological control agents. The life histories and host ranges of these beetles were studied under laboratory conditions to determine their suitability for release. The two beetle species displayed very similar life histories, including a short pre-oviposition period (13–14 days), incubation period (4–5 days) and lifecycle (40–45 days). The longevity of Z. signatipennis and Z. piceicollis was 113 and 125 days, while their fecundities were 1146 and 1133 eggs per female, respectively. Feeding, oviposition and development of both beetle species were confined within the tribe Heliantheae, but showed a very strong preference for the invasive T. rotundifolia. The only non-target species that supported development to adulthood was the exotic weed Tithonia diversifolia (Hemsl.) A. Gray, itself a target for biocontrol. Although some sunflower (Helianthus annuus L.) cultivars were partially utilized during host-specificity tests, none supported complete development, suggesting that both Zygogramma species are suitable for release in South Africa.

Selection of the Most Effective Trichogramma Strains (Hymenoptera: Trichogrammatidae) From Myanmar to Control Asian Corn Borer, Ostrinia furnacalis (Lepidoptera: Crambidae).

Selecting the candidate agents or species is a fundamental step in developing effective biological control programs. However, no attempts have been made to evaluate the efficacy of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) strains against the Asian corn borer, Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) in Myanmar. In this study, Trichogramma strains were investigated through a series of glass tube bioassays and under a semifield condition. Ostrinia furnacalis or Corcyra cephalonica egg preferences were compared among six strains of Trichogramma ostriniae, three of Trichogramma chilonis, and four of Trichogramma dendrolimi using a choice-test assay design. Significant differences were observed at the inter- and intraspecific levels. Four strains of T. ostriniae and two of T. dendrolimi showed a strong preference for O. furnacalis eggs, while two strains of T. chilonis and one T. dendrolimi strain preferred Corcyra cephalonica Stainton (Lepidoptera: Pyralidae) eggs. The remaining strains showed no preference. Eleven strains were examined on O. furnacalis eggs only, wherein parasitism, sex ratio, and total progeny per female were highest for two strains of T. ostriniae (respectively: 67.6 ± 3.0%, 82.7 ± 2.3%, 49.6 ± 2.8, and 67.6 ± 3.6%, 90.0 ± 2.4%, 42.7 ± 2.6), and one strain of T. dendrolimi (65.2 ± 3.2%, 84.7 ± 2.6%, 46.3 ± 2.8). Parasitism and searching capacity were evaluated for the three, best performing strains in cages containing maize plants. One T. ostriniae from Southern Shan State, Myanmar parasitized ~81% of egg masses, and should be considered a candidate biological control agent against O. furnacalis in Myanmar.

An innovative approach combining metabarcoding and ecological interaction networks for selecting candidate biological control agents

AbstractClassical biological control (CBC) can be used to decrease the density of invasive species to below an acceptable ecological and economic threshold. Natural enemies specific to the invasive species are selected from its native range and released into the invaded range. This approach has drawbacks, despite the performance of specificity tests to ensure its safety, because the fundamental host range defined under controlled conditions does not represent the actual host rangein natura, and these tests omit indirect interactions within community.We focus onSonchus oleraceus(Asteraceae), a weed species originating from Western Palearctic that is invasive in many countries and notably in Australia. We explore how analyses of interaction network within its native range can be used to (a) inventory herbivores associated to the target plant, (b) characterize their ecological host ranges and (c) guide the selection of candidate biocontrol agents considering interactions with species from higher trophic levels. Arthropods were collected from plant community sympatric toS. oleraceus, in three bioclimatic regions, and interactions were inferred by a combination of molecular and morphological approaches.The networks reconstructed were structured in several trophic levels from basal species (plant community), to intermediate and top species (herbivorous arthropods and their natural enemies). The sub‐network centred onS. oleraceus‐related interactions contained 116 taxa and 213 interactions. We identified 47 herbivores feeding onS. oleraceus, 15 of which were specific to the target species. Some discrepancies with respect to published findings or conventional specificity tests suggested possible insufficient sampling effort for the recording of interactions or the existence of cryptic species. Among potential candidate agents, six exhibited interactions with natural enemies.Synthesis and applications. Adopting a network approach as prerequisite step of the classical biological control programme can provide a rapid screening of potential agents to be tested in priority. Once ecological host range defined, we suggest that priority should be given to agent used by a minimum species, and, when they exist, to agents that possess enemies from the most distant taxonomical group from those occurring in the range of introduction.

Physiological host range of Trissolcus mitsukurii, a candidate biological control agent of Halyomorpha halys in Europe

The invasive stink bug Halyomorpha halys, native to East Asia, is a severe agricultural pest of worldwide importance, and chemical insecticides are largely sprayed for its control. Negative impact and failures of chemical pest management led to consider classical biological control as one of the most promising methods in a long-term perspective. The Asian egg parasitoid Trissolcus japonicus is the main candidate biocontrol agent of H. halys, but more recently a second species, Trissolcus mitsukurii, which shares the area of origin with H. halys, has drawn special attention after adventive populations were found in Europe. Despite its recent detection, intentional release or redistribution are bound to approval of national petitions, which requires detailed risk analyses. With the aim of providing a contribution for the future development of a risk assessment for release purposes, here we present the first study on the physiological (fundamental) host range of T. mitsukurii in Europe. Tests conducted on different hemipterans common to Southern Europe, using three different experimental designs, revealed a oligophagous host range, limited to Pentatomoidea and comparable with the host range displayed by T. japonicus under similar laboratory conditions. In addition to its coevolved host H. halys, T. mitsukurii successfully parasitized the majority of tested pentatomid species and one scutellerid, although with highly variable emergence rates. Notably high parasitization rates were detected on H. halys, Acrosternum heegeri and Dolycoris baccarum. Host egg sizes positively affected parasitoid size and female egg load. Although T. mitsukurii might be a promising biological control agent of H. halys in Europe, further studies (e.g., chemical ecology, field parasitism) are needed for assessment of non-target risks.

Revealing the Introduction History and Phylogenetic Relationships of Passiflora foetida sensu lato in Australia.

Genomic analysis can be a valuable tool to assistmanagement of non-native invasive species, through determining source and number of introductions as well as clarifying phylogenetic relationships. Here, we used whole chloroplast sequencing to investigate the introduction history of Passiflora foetida sensu lato in Australia and clarify its relationship with other Passiflora species present. Phylogenetic analysis of chloroplast genome data identified three separate genetic lineages of P. foetida s. l. present in Australia, indicating multiple introductions. These lineages had affinities to samples from three separate areas within the native range in Central and South America that represented phylogenetically distinct lineages. These results provide a basis for a targeted search of the native range of P. foetida s. l. for candidate biological control agents that have co-evolved with this species and are thus better adapted to the lineages that are present in Australia. Results also indicated that the Passiflora species native to Australia are in a separate clade to that of P. foetida s. l. and other introduced Passiflora species cultivated in Australia. This knowledge is important to assess the likelihood of finding biological control agents for P. foetida s. l. that will be sufficiently host-specific for introduction in Australia. As P. foetida s. l. is a widespread non-native invasive species across many regions of the world, outcomes from this work highlight the importance of first evaluating the specific entities present in a country before the initiation of a biological control program.