'Candidatus Liberibacter brunswickensis' colonization has no effect to the early development of Solanum melongena.

This study is the first to investigate the presence and movement of the novel Liberibacter species 'Candidatus Liberibacter brunswickensis' (CLbr) in eggplant, Solanum melongena. The psyllid, Acizzia solanicola can transmit CLbr to eggplant and CLbr can be acquired by CLbr-negative A. solanicola individuals from CLbr-positive eggplants. In planta, CLbr can replicate, move and persist. Investigation into the early development of eggplants showed that CLbr titres had increased at the inoculation site at 14 days post inoculation access period (DPIAP). CLbr had become systemic in the majority of plants tested by 28 DPIAP. The highest bacterial titres were recorded at 35 DPIAP in all samples of the inoculated leaf, the roots, stems and the midrib and petiole samples of the newest leaf (the top leaf). This finding strongly suggests that CLbr movement in planta follows the source to sink relationship as previously described for 'Ca. Liberibacter asiaticus' (CLas) and 'Ca. Liberibacter solanacearum' (CLso). No symptoms consistent with Liberibacter-associated diseases were noted for plants colonised by CLbr during this study, consistent with the hypothesis that CLbr does not cause disease of eggplant during the early stages of host colonisation. In addition, no significant differences in biomass were found between eggplant colonised with CLbr, compared to those that were exposed to CLbr-negative A. solanicola, and to control plants.

Two independent LAMP assays for rapid identification of the serpentine leafminer, Liriomyza huidobrensis (Blanchard, 1926) (Diptera: Agromyzidae) in Australia.

Liriomyza huidobrensis is a leafminer fly and significant horticultural pest. It is a quarantine listed species in many countries and is now present as an established pest in Australia. Liriomyza huidobrensis uses a broad range of host plants and has potential for spread into various horticultural systems and regions of Australia. Rapid in-field identification of the pest is critically needed to assist efforts to manage this pest. Morphological identification of the pest is effectively limited to specialist examinations of adult males. Generally, molecular methods such as qPCR and DNA barcoding for identification of Liriomyza species require costly laboratory-based hardware. Herein, we developed two independent and rapid LAMP assays targeted to independently inherited mitochondrial and nuclear genes. Both assays are highly sensitive and specific to L. huidobrensis. Positive signals can be detected within 10 min on laboratory and portable real-time amplification fluorometers. Further, we adapted these assays for use with colorimetric master mixes, to allow fluorometer free in-field diagnostics of L. huidobrensis. Our LAMP assays can be used for stand-alone testing of query specimens and are likely to be essential tools used for rapid identification and monitoring of L. huidobrensis.

Molecular identification of hymenopteran parasitoids and their endosymbionts from agromyzids.

Three polyphagous pest Liriomyza spp. (Diptera: Agromyzidae) have recently invaded Australia and are damaging horticultural crops. Parasitic wasps are recognized as effective natural enemies of leafmining species globally and are expected to become important biocontrol agents in Australia. However, the hymenopteran parasitoid complex of agromyzids in Australia is poorly known and its use hindered due to taxonomic challenges when based on morphological characters. Here, we identified 14 parasitoid species of leafminers based on molecular and morphological data. We linked DNA barcodes (5' end cytochrome c oxidase subunit I (COI) sequences) to five adventive eulophid wasp species (Chrysocharis pubicornis (Zetterstedt), Diglyphus isaea (Walker), Hemiptarsenus varicornis (Girault), Neochrysocharis formosa (Westwood), and Neochrysocharis okazakii Kamijo) and two braconid species (Dacnusa areolaris (Nees) and Opius cinerariae Fischer). We also provide the first DNA barcodes (5' end COI sequences) with linked morphological characters for seven wasp species, with three identified to species level (Closterocerus mirabilis Edwards & La Salle, Trigonogastrella parasitica (Girault), and Zagrammosoma latilineatum Ubaidillah) and four identified to genus (Aprostocetus sp., Asecodes sp., Opius sp. 1, and Opius sp. 2). Phylogenetic analyses suggest C. pubicornis, D. isaea, H. varicornis, and O. cinerariae are likely cryptic species complexes. Neochrysocharis formosa and Aprostocetus sp. specimens were infected with Rickettsia. Five other species (Cl. mirabilis, D. isaea, H. varicornis, Opius sp. 1, and Opius sp. 2) were infected with Wolbachia, while two endosymbionts (Rickettsia and Wolbachia) co-infected N. okazakii. These findings provide background information about the parasitoid fauna expected to help control the leafminers.

Meschia brevirostris sp. nov., from New Caledonia (Hemiptera: Heteroptera: Lygaeoidea: Meschiidae).

A new species, Meschia brevirostris sp. nov. (Hemiptera: Heteroptera: Lygaeoidea: Meschiidae), is described from New Caledonia. Photographs and SEM micrographs of the male and female habitus, genital structures and selected morphological structures are presented.

Revision of a unique Australian leafhopper genus Stenopsoides Evans (Hemiptera: Cicadellidae: Idiocerinae: Macropsini).

The Australian genus Stenopsoides Evans (Idiocerinae: Macropsini) is revised. The type species, S. turneri Evans, is redescribed and three new species are added: S. newi Semeraro sp. nov., S. punctatus Semeraro sp. nov. and S. truncatus Semeraro sp. nov.. Species photographic plates, a distribution map and an illustrated key to the four species are provided. The possibility of parthenogenetic reproduction occuring in this genus is discussed.

Indigenous plants promote insect biodiversity in urban greenspaces.

The contribution of urban greenspaces to support biodiversity and provide benefits for people is increasingly recognized. However, ongoing management practices favor vegetation oversimplification, often limiting greenspaces to lawns and tree canopy rather than multi-layered vegetation that includes under- and midstorey, and the use of nonnative species. These practices hinder the potential of greenspaces to sustain indigenous biodiversity, particularly for taxa like insects that rely on plants for food and habitat. Yet, little is known about which plant species may maximize positive outcomes for taxonomically and functionally diverse insect communities in greenspaces. Additionally, while cities are expected to experience high rates of introductions, quantitative assessments of the relative occupancy of indigenous vs. introduced insect species in greenspace are rare, hindering understanding of how management may promote indigenous biodiversity while limiting the establishment of introduced insects. Using a hierarchically replicated study design across 15 public parks, we recorded occurrence data from 552 insect species on 133 plant species, differing in planting design element (lawn, midstorey, and tree canopy), midstorey growth form (forbs, lilioids, graminoids, and shrubs) and origin (nonnative, native, and indigenous), to assess (1) the relative contributions of indigenous and introduced insect species and (2) which plant species sustained the highest number of indigenous insects. We found that the insect community was overwhelmingly composed of indigenous rather than introduced species. Our findings further highlight the core role of multi-layered vegetation in sustaining high insect biodiversity in urban areas, with indigenous midstorey and canopy representing key elements to maintain rich and functionally diverse indigenous insect communities. Intriguingly, graminoids supported the highest indigenous insect richness across all studied growth forms by plant origin groups. Our work highlights the opportunity presented by indigenous understory and midstorey plants, particularly indigenous graminoids, in our study area to promote indigenous insect biodiversity in urban greenspaces. Our study provides a blueprint and stimulus for architects, engineers, developers, designers, and planners to incorporate into their practice plant species palettes that foster a larger presence of indigenous over regionally native or nonnative plant species, while incorporating a broader mixture of midstorey growth forms.

Novel bioassay to assess antibiotic effects of fungal endophytes on aphids.

Perennial ryegrass is an important feed base for the dairy and livestock industries around the world. It is often infected with mutualistic fungal endophytes that confer protection to the plant against biotic and abiotic stresses. Bioassays that test their antibiotic effect on invertebrates are varied and range from excised leaves to whole plants. The aim of this study was to design and validate a "high-throughput" in-planta bioassay using 7-day-old seedlings confined in small cups, allowing for rapid assessments of aphid life history to be made while maintaining high replication and treatment numbers. Antibiosis was evaluated on the foliar and the root aphid species; Diuraphis noxia (Mordvilko) and Aploneura lentisci (Passerini) feeding on a range of perennial ryegrass-Epichloë festucae var. Lolii endophyte symbiota. As expected, both D. noxia and A. lentisci reared on endophyte-infected plants showed negatively affected life history traits by comparison to non-infected plants. Both species exhibited the highest mortality at the nymphal stage with an average total mortality across all endophyte treatments of 91% and 89% for D. noxia and A. lentisci respectively. Fecundity decreased significantly on all endophyte treatments with an average total reduction of 18% and 16% for D. noxia and A. lentisci respectively by comparison to non-infected plants. Overall, the bioassay proved to be a rapid method of evaluating the insecticidal activity of perennial ryegrass-endophyte symbiota on aphids (nymph mortality could be assessed in as little as 24 and 48 hours for D. noxia and A. lentisci respectively). This rapid and simple approach can be used to benchmark novel grass-endophyte symbiota on a range of aphid species that feed on leaves of plants, however we would caution that it may not be suitable for the assessment of root-feeding aphids, as this species exhibited relatively high mortality on the control as well.

Phylogenetic analysis of the Australasian paralysis ticks and their relatives (Ixodidae: Ixodes: Sternalixodes).

BACKGROUND: The Australasian paralysis ticks and their relatives, Ixodes Latrielle, subgenus Sternalixodes Schulze, are some of the most important ticks in the region. However, very little is known about their phylogenetic relationships. The aim of this study was to elucidate the evolutionary relationships of members of the subgenus Sternalixodes by undertaking phylogenetic analyses of morphological and molecular datasets. METHODS: Adult females (n = 64) of Sternalixodes, including Ixodes anatis Chilton, 1904, Ixodes confusus Roberts, 1960, Ixodes cornuatus Roberts, 1960, Ixodes cordifer Neumann, 1908, Ixodes dendrolagi Wilson, 1967, Ixodes hirsti Hassall, 1931, Ixodes holocyclus Neumann, 1899, Ixodes myrmecobii Roberts, 1962 and Ixodes trichosuri Roberts, 1960, were examined morphologically. Subsequently, these Ixodes spp. were genetically characterised using cytochrome c oxidase subunit 1 (cox1) gene and the internal transcribed spacer 2 (ITS-2) of the rRNA. Both morphological and molecular datasets were analysed using various phylogenetic methods to assess the evolutionary relationship of various members of the subgenus Sternalixodes. RESULTS: Phylogenetic analyses of the cox1 sequences and morphological characters datasets revealed that the Australian and Papuan Sternalixodes formed a distinct clade with the New Zealand member of the group I. anatis positioned basally, in a separate clade. Ixodes holocyclus, I. cornuatus and I. myrmecobii formed a distinctive clade in both the cox1 and morphological phylogenies. However, based on phylogenetic analysis of the ITS-2 data, I. holocyclus formed a separate clade whereas I. cornuatus and I. myrmecobii grouped in a different clade. CONCLUSIONS: The cox1 and morphological data suggest that the subgenus Sternalixodes is paraphyletic, and I. anatis is not a sternalixodid tick; hence, it should not be included in the subgenus. Based on the phylogenetic analyses of cox1 and ITS-2 sequences, it appears that I. myrmecobii and I. cornuatus are not subspecies of I. holocyclus. Although this study provided better insights into the taxonomic status of the subgenus Sternalixodes, a complete morphological and molecular (using multiple markers) phylogenetic analysis including all members of the subgenus would be required to more accurately elucidate the evolutionary relationships within the subgenus.

Conserving herbivorous and predatory insects in urban green spaces.

Insects are key components of urban ecological networks and are greatly impacted by anthropogenic activities. Yet, few studies have examined how insect functional groups respond to changes to urban vegetation associated with different management actions. We investigated the response of herbivorous and predatory heteropteran bugs to differences in vegetation structure and diversity in golf courses, gardens and parks. We assessed how the species richness of these groups varied amongst green space types, and the effect of vegetation volume and plant diversity on trophic- and species-specific occupancy. We found that golf courses sustain higher species richness of herbivores and predators than parks and gardens. At the trophic- and species-specific levels, herbivores and predators show strong positive responses to vegetation volume. The effect of plant diversity, however, is distinctly species-specific, with species showing both positive and negative responses. Our findings further suggest that high occupancy of bugs is obtained in green spaces with specific combinations of vegetation structure and diversity. The challenge for managers is to boost green space conservation value through actions promoting synergistic combinations of vegetation structure and diversity. Tackling this conservation challenge could provide enormous benefits for other elements of urban ecological networks and people that live in cities.

Barcoding Queensland Fruit Flies (Bactrocera tryoni): impediments and improvements.

Identification of adult fruit flies primarily involves microscopic examination of diagnostic morphological characters, while immature stages, such as larvae, can be more problematic. One of the Australia's most serious horticultural pests, the Queensland Fruit Fly (Bactrocera tryoni: Tephritidae), is of particular biosecurity/quarantine concern as the immature life stages occur within food produce and can be difficult to identify using morphological characteristics. DNA barcoding of the mitochondrial Cytochrome Oxidase I (COI) gene could be employed to increase the accuracy of fruit fly species identifications. In our study, we tested the utility of standard DNA barcoding techniques and found them to be problematic for Queensland Fruit Flies, which (i) possess a nuclear copy (a numt pseudogene) of the barcoding region of COI that can be co-amplified; and (ii) as in previous COI phylogenetic analyses closely related B. tryoni complex species appear polyphyletic. We found that the presence of a large deletion in the numt copy of COI allowed an alternative primer to be designed to only amplify the mitochondrial COI locus in tephritid fruit flies. Comparisons of alternative commonly utilized mitochondrial genes, Cytochrome Oxidase II and Cytochrome b, revealed a similar level of variation to COI; however, COI is the most informative for DNA barcoding, given the large number of sequences from other tephritid fruit fly species available for comparison. Adopting DNA barcoding for the identification of problematic fly specimens provides a powerful tool to distinguish serious quarantine fruit fly pests (Tephritidae) from endemic fly species of lesser concern.