Thynnine Wasp Research Articles

Resilience to summer bushfire in the threatened orchid, Caladenia tessellata, in terms of pollination success, herbivory, and mycorrhizal associations

Abstract In some biogeographic regions, many threatened plant species occur in habitats that periodically experience bushfire. However, we currently have relatively little information on how important plant–animal and plant–fungus interactions are affected by these fires. For the threatened sexually deceptive orchid Caladenia tessellata we test whether pollination rate, frequency of florivory, and the species of mycorrhizae the plant associates with differ between burnt and unburnt sites. Interestingly, pollination rates were unaffected by fire, demonstrating that populations of the thynnine wasp pollinator can persist post-fire. However, there was a significant negative relationship between number of flowers in a population and pollination success, which is likely a by-product of a deceptive pollination strategy. Despite the presence of vertebrate herbivores, florivory rates were low in both burnt and unburnt sites. Caladenia tessellata associated primarily with the mycorrhiza Serendiptia australiana, regardless of fire history. While our results suggest resilience to a one-off summer fire for the ecological interactions that we measured, it would be interesting to investigate the effects of fire frequency and time of year. High reproductive rates in small populations of C. tessellata suggest these populations may be viable and that retaining them is a high priority for conservation.

Improving conservation and translocation success of an endangered orchid, Caladenia xanthochila (Orchidaceae), through understanding pollination

Critical for conserving endangered orchids is identifying their pollinators and their distribution. Caladenia xanthochila is an endangered orchid that has floral traits characteristic of pollination by food foraging insects. We identified the pollinator(s), mechanisms of attraction and the presence of pollinators at natural, existing and potential translocation sites. Furthermore, we quantified pollination success at translocation sites and investigated the effect of rainfall on pollination success over 19 years at a natural site. We clarify if sharing of pollinators occurs with closely related species by comparing the CO1 barcoding region of the pollinators' DNA. Caladenia xanthochila was pollinated by a single species of thynnine wasp, Phymatothynnus aff. nitidus. Caladenia xanthochila produced 27.0 µg ± 7.1 sucrose on the labellum, while pollinators vigorously copulated with glandular clubs on the sepal tips, suggestive of a mixed pollination system. Pollination success of C. xanthochila was 7.6 ± 1.5% SE at the natural site and 16.1 ± 3.6% SE across the translocation sites. Furthermore, hand pollinations demonstrated that pollination was pollen limited. Pollination success was significantly related to average rainfall during the growth phase of the orchid (P < 0.001). Potential translocation sites for C. xanthochila were limited, with four of six surveyed lacking the pollinator. We found evidence for cryptic species of Phymatothynnus, with C. xanthochila pollinators being unique amongst the orchids studied. We recommend hand pollinations at translocated and remnant wild populations to boost initial recruitment. The evidence for cryptic species of pollinators further highlights the need for accurate identification of pollinators.

Drakolide Structure-activity Relationships for Sexual Attraction of Zeleboria Wasp Pollinator.

Orchids pollinated by sexual deception lure their specific male pollinators by sex pheromone mimicry. Despite the growing list of chemically diverse semiochemicals known to be involved, the chemical basis and flexibility of this extreme pollinator specificity are not fully understood. One promising but rarely applied tool is the synthesis and field testing of chemically related variants for investigating the structural specificity of the pheromone mimics. Here, we build on the discovery of the unusual semiochemical blend used by Drakaea micrantha to sexually lure its male Zeleboria thynnine wasp pollinator. This blend consists of a β-ketolactone (drakolide) and two specific hydroxymethylpyrazines, presumably drawn from two distinct biosynthetic pathways. Here, we synthesized and tested the activity of various stereo- and structural isomers of the naturally occurring drakolide. Our study confirmed that in blends with the two pyrazines, both a mixture of stereoisomers, and the specific stereoisomer of the natural drakolide, elicit high rates of landings and attempted copulations. However, in the absence of pyrazines, both the number of responses and the level of sexual attraction were significantly reduced. When structural analogs were substituted for the natural drakolide, attractiveness and degree of sexual behaviour varied but were generally reduced. Based on our findings, and prior knowledge that related hydroxymethylpyrazines are active in other Drakaea spp., we conclude that the dual sex pheromone mimicry of D. micrantha likely evolved via initial changes in just one of the two biosynthetic pathways. Most plausibly, this involved modifications in the drakolides, with the pyrazines as a 'pre-adaption' enhancing the sexual response.

Three Chemically Distinct Floral Ecotypes in Drakaea livida, an Orchid Pollinated by Sexual Deception of Thynnine Wasps.

Sexually deceptive orchids are unusual among plants in that closely related species typically attract different pollinator species using contrasting blends of floral volatiles. Therefore, intraspecific variation in pollinator attraction may also be underpinned by differences in floral volatiles. Here, we tested for the presence of floral ecotypes in the sexually deceptive orchid Drakaea livida and investigated if the geographic range of floral ecotypes corresponded to variation in pollinator availability. Pollinator choice trials revealed the presence of three floral ecotypes within D. livida that each attracts a different species of thynnine wasp as a pollinator. Surveys of pollinator distribution revealed that the distribution of one of the ecotypes was strongly correlated with that of its pollinator, while another pollinator species was present throughout the range of all three ecotypes, demonstrating that pollinator availability does not always correlate with ecotype distribution. Floral ecotypes differed in chemical volatile composition, with a high degree of separation evident in principal coordinate analysis. Some compounds that differed between ecotypes, including pyrazines and (methylthio)phenols, are known to be electrophysiologically active in thynnine wasp antennae. Based on differences in pollinator response and floral volatile profile, the ecotypes represent distinct entities and should be treated as such in conservation management.

Evolution of reproductive structures for in-flight mating in thynnine wasps (Hymenoptera: Thynnidae: Thynninae).

Thynnine wasps have an unusual mating system that involves concurrent in-flight copulation and nuptial feeding of wingless females by alate males. Consequently, thynnine genitalia play a multifunctional role and have likely been subject to various different selective pressures for both reproductive success and food provisioning. Here, we present a new molecular phylogeny for the Australian Thynninae and use 3D-geometric morphometrics and comparative methods to investigate the morphological evolution of select genital structures across the group. We found significant morphological integration between all male and female structures analysed, which is likely influenced by sexual selection, but also reproductive isolation requirements and mechanical constraints. The morphology of the primary male and female coupling structures was correlated with female body size, and female genitalia exhibited strong negative size allometry. Those male and female coupling structures have evolved at similar evolutionary rates, whereas female structures appear to have evolved a higher degree of morphological novelty over time. We conclude that the unique reproductive strategies of thynnine wasps have resulted in complex evolutionary patterns in their genital morphology, which has likely played a central role in the extensive diversification of the subfamily across Australasia and South America. Our study reinforces the need to treat composite characters such as genitalia by their component parts, and to consider the roles of both male and female reproductive structures in evolutionary studies.

A specialised pollination system using nectar-seeking thynnine wasps in Caladenia nobilis (Orchidaceae).

Caladenia is a diverse Australian genus that is exceptional among orchids in having both species pollinated by food-seeking and sexually deceived insects. Here, we investigated the pollination of Caladenia nobilis, a species predicted to be food-deceptive due to its large, cream-coloured and apparently nectarless flowers. Pollinator observations were made using experimental clumps of flowers. Measurements of floral colour were undertaken with a spectrometer, nectar was tested using GC-MS, and reproductive success was quantified for 2years. While C.nobilis attracted nine species of insect, only males of the thynnine wasp Rhagigaster discrepans exhibited the correct size and behaviour to remove and deposit pollen. Male R.discrepans attempted to feed from the surface of the labellum, often crawling to multiple flowers, but showed no evidence of sexual attraction. Most flowers produced little or no nectar, although some may provide enough sucrose to act as a meagre reward to pollinators. Floral colouration was similar to a related Caladenia species pollinated by sexual deception, although the sexually deceptive species had a dull-red labellum. Reproductive success was generally low and highly variable between sites and years. In addition to most visitors being of inappropriate size for pollinia removal, the lack of response to the orchid by several co-occurring species of thynnine wasp suggests filtering of potential pollinators at the attraction phase. Our discovery of a pollination strategy that may be intermediate between food deception and food reward raises the question, how many putatively rewardless orchids actually produce meagre amounts of nectar?

Pollination by nectar-foraging thynnine wasps in the endangered Caladenia arenaria and Caladenia concolor (Orchidaceae)

Prior to undertaking conservation translocations of plants with specialised pollination systems, it is important to ensure the presence of pollinators at recipient sites. Here, for two threatened species, Caladenia concolor Fitzg. and Caladenia arenaria Fitzg. (Orchidaceae), we determine (i) the pollination strategy used, (ii) which floral visitors are involved in pollination, and (iii) whether the pollinator species are present at potential translocation sites. For both orchid species, pollination was primarily achieved by nectar-foraging thynnine wasps, with a single species responsible for pollination in C. concolor, whereas C. arenaria utilised at least two species to achieve pollination. Both orchid species secreted meagre quantities of sucrose on the upper surface of the labellum. Visits to C. concolor occurred primarily in the late afternoon, with some wasps perching on the flowers overnight. Surveys revealed that pollinators were present at all extant populations and most potential translocation sites for both orchids. The specialisation on one pollinator species in C. concolor means that the distribution of the pollinator needs to be considered for conservation translocations. With C. arenaria, the risk of hybridisation with other Caladenia that are known to share one of its pollinator species needs to be taken into account when selecting translocation sites.

Pollination by nectar-foraging thynnine wasps: evidence of a new specialized pollination system for Australian orchids

This work was supported by funding to Noushka Reiter through a Victorian Threatened Species Initiative Grant and National Landcare Programme funding through the Wimmera Catchment Management Authority. Ryan Phillips and Bjorn Bohman were supported by Australian Research Council Discovery Early Career Research Awards (DE15010720 and DE160101313 respectively).

Structure-Activity Studies of Semiochemicals from the Spider Orchid Caladenia plicata for Sexual Deception.

Sexually deceptive orchids attract specific pollinators by mimicking insect sex pheromones. Normally this mimicry is very specific and identical compounds have been identified from orchids and matching females of the pollinators. In this study, we conduct a detailed structure-activity investigation on isomers of the semiochemicals involved in the sexual attraction of the male pollinator of the spider orchid Caladenia plicata. This orchid employs an unusual blend of two biosynthetically unrelated compounds, (S)-β-citronellol and 2-hydroxy-6-methylacetophenone, to lure its Zeleboria sp. thynnine wasp pollinator. We show that the blend is barely attractive when (S)-β-citronellol is substituted with its enantiomer, (R)-β-citronellol. Furthermore, none of the nine-possible alternative hydroxy-methylacetophenone regioisomers of the natural semiochemical are active when substituted for the natural 2-hydroxy-6-methylacetophenone. Our results were surprising given the structural similarity between the active compound and some of the analogues tested, and results from previous studies in other sexually deceptive orchid/wasp systems where substitution with analogues was possible. Interestingly, high-level ab initio and density functional theory calculations of the hydroxy-methylacetophenones revealed that the active natural isomer, 2-hydroxy-6-methylacetophenone, has the strongest intramolecular hydrogen bond of all regioisomers, which at least in part may explain the specific activity.

Breaking the rules: discovery of sexual deception in Caladenia abbreviata (Orchidaceae), a species with brightly coloured flowers and a non-insectiform labellum

Caladenia is exceptional among orchid genera in that it contains both species pollinated by sexual deception and species pollinated by food-foraging insects. Whereas pollination strategies have been elucidated in some species complexes, others groups have received limited attention, hampering our ability to understand the evolution of the exceptional diversity of species and floral traits in Caladenia. Here, we conducted the first detailed pollination study of a member of the Caladenia filamentosa complex. On the basis of the brightly coloured and scented flowers exhibited by many species, it has been assumed that most members of the group are food-deceptive. However, we show that Caladenia abbreviata Hopper &amp; A.P.Br. is pollinated by an undescribed species of Rhytidothynnus thynnine wasp via sexual deception. Floral dissections showed that the terminal glands on the sepals and petals are the primary source of the sexual attractant, supporting observations from intact flowers that pollinators frequently copulate with these floral parts. Interestingly, contact with the reproductive structures of the plant was not associated with attempted copulation. Our findings highlighted that sexual deception can evolve in lineages without dull-coloured flowers and insectiform labellum structures. Given that the orchid is apparently dependent on a single pollinator species, efforts to conserve this rare species will be dependent on the availability of the pollinator in suitable orchid habitat.

(Methylthio)phenol semiochemicals are exploited by deceptive orchids as sexual attractants for Campylothynnus thynnine wasps

Until recently, (methylthio)phenols as natural products had only been reported from bacteria. Now, four representatives of this class of sulfurous aromatic compounds have been discovered as semiochemicals in the orchid Caladenia crebra, which secures pollination by sexual deception. In this case, field bioassays confirmed that a 10:1 blend of 2-(methylthio)benzene-1,4-diol (1) and 4-hydroxy-3-(methylthio)benzaldehyde (2) sexually attracts the male thynnine wasp Campylothynnus flavopictus (Tiphiidae:Thynnineae), the exclusive pollinator of C. crebra. Here we show with field bioassays that another undescribed species of Campylothynnus (sp. A) is strongly sexually attracted to a 1:1 blend of compounds 1 and 2, which elicits very high attempted copulation rates (88%). We also confirm that this Campylothynnus species is a pollinator of Caladenia attingens subsp. attingens. Chemical analysis of the flowers of this orchid revealed two (methylthio)phenols, compound 2 and 2-(methylthio)phenol (3), as candidate semiochemicals involved in pollinator attraction. Thus, (methylthio)phenols are likely to be more widely used than presently known. The confirmation of this Campylothynnus as a pollinator of C. attingens subsp. attingens at our study sites was unexpected, since elsewhere this orchid is pollinated by a different thynnine wasp (Thynnoides sp). In general, sexually deceptive Caladenia only use a single species of pollinator, and as such, this unusual case may offer a tractable study system for understanding the chemical basis of pollinator switching in sexually deceptive orchids.

Evolutionary relationships among pollinators and repeated pollinator sharing in sexually deceptive orchids.

The mechanism of pollinator attraction is predicted to strongly influence both plant diversification and the extent of pollinator sharing between species. Sexually deceptive orchids rely on mimicry of species-specific sex pheromones to attract their insect pollinators. Given that sex pheromones tend to be conserved among related species, we predicted that in sexually deceptive orchids, (i) pollinator sharing is rare, (ii) closely related orchids use closely related pollinators and (iii) there is strong bias in the wasp lineages exploited by orchids. We focused on species that are pollinated by sexual deception of thynnine wasps in the distantly related genera Caladenia and Drakaea, including new field observations for 45 species of Caladenia. Specialization was extreme with most orchids using a single pollinator species. Unexpectedly, seven cases of pollinator sharing were found, including two between Caladenia and Drakaea, which exhibit strikingly different floral morphology. Phylogenetic analysis of pollinators using four nuclear sequence loci demonstrated that although orchids within major clades primarily use closely related pollinator species, up to 17% of orchids within these clades are pollinated by a member of a phylogenetically distant wasp genus. Further, compared to the total diversity of thynnine wasps within the study region, orchids show a strong bias towards exploiting certain genera. Although these patterns may arise through conservatism in the chemical classes used in sex pheromones, apparent switches between wasp clades suggest unexpected flexibility in floral semiochemical production. Alternatively, wasp sex pheromones within lineages may exhibit greater chemical diversity than currently appreciated.

Complex Sexual Deception in an Orchid Is Achieved by Co-opting Two Independent Biosynthetic Pathways for Pollinator Attraction

Sexually deceptive orchids lure their specific male pollinators using volatile semiochemicals that mimic female sex pheromones. To date, the semiochemicals known to be involved consist of blends of chemically and biosynthetically related compounds. In contrast, we report that (S)-β-citronellol and 2-hydroxy-6-methylacetophenone, two biosynthetically distinct compounds, are the active semiochemicals in Caladenia plicata, which is pollinated by male Zeleboria sp. thynnine wasps. They are also sex pheromone components of the female Zeleboria. A1:4 blend elicits a high rate of attempted copulation (∼70%) in bioassays, equivalent to rates observed at orchid flowers. Whereas β-citronellol is well known, 2-hydroxy-6-methylacetophenone appears to be previously unknown as a floral volatile. Production of the two compounds is restricted to glandular sepal tips; thus, differential expression analysis of contrasting floral tissue transcriptomes was employed to illuminate the biosynthesis. As expected, production of (S)-β-citronellol commences with the terpene synthase GES1 catalyzing the irreversible conversion of geranyl diphosphate (GPP) to geraniol. Contrary to prediction, biosynthesis subsequently proceeds in three steps, commencing with the oxidation of geraniol to geranial by alcohol dehydrogenase ADH3, followed by the enantioselective reduction of a double bond in geranial by geranial reductase GER1 to give (S)-β-citronellal. Finally, ADH3-catalyzed reduction of (S)-β-citronellal results in (S)-β-citronellol. Inline with previous work on insects showing that 2-hydroxy-6-methylacetophenone is derived from a polyketide pathway, we report a differentially expressed polyketide synthase (PKS) gene candidate. Thus, in this unique example of sexual deception, pollination is achieved by co-opting and regulating two independent biosynthetic pathways of floral volatile compounds. VIDEO ABSTRACT.

The Spider Orchid Caladenia crebra Produces Sulfurous Pheromone Mimics to Attract its Male Wasp Pollinator

One of the most intriguing natural observations is the pollination of orchids by sexual deception. Chemicals underpin this interaction between the orchid and its sexually attracted male insect pollinator, with the signaling compounds involved, called semiochemicals, predicted to mimic the chemical composition of the sex pheromone. We identified floral semiochemicals from Caladenia (spider orchids) for the first time. We further demonstrate that C. crebra attracts its single pollinator species with a unique system of (methylthio)phenols, three of which are new natural products. Furthermore, as predicted, the same compounds constitute the sex pheromone of the pollinator, the thynnine wasp Campylothynnus flavopictus, representing the first occurrence of sulfurous sex pheromones in Hymenoptera.

The Spider Orchid Caladenia crebra Produces Sulfurous Pheromone Mimics to Attract its Male Wasp Pollinator

AbstractOne of the most intriguing natural observations is the pollination of orchids by sexual deception. Chemicals underpin this interaction between the orchid and its sexually attracted male insect pollinator, with the signaling compounds involved, called semiochemicals, predicted to mimic the chemical composition of the sex pheromone. We identified floral semiochemicals from Caladenia (spider orchids) for the first time. We further demonstrate that C. crebra attracts its single pollinator species with a unique system of (methylthio)phenols, three of which are new natural products. Furthermore, as predicted, the same compounds constitute the sex pheromone of the pollinator, the thynnine wasp Campylothynnus flavopictus, representing the first occurrence of sulfurous sex pheromones in Hymenoptera.

Fire, food and sexual deception in the neighbourhood of some Australian orchids

AbstractThe effective use of prescribed fire in biodiversity conservation is currently inhibited by a limited understanding of fire effects on ecosystem processes such as pollination. Orchids inhabiting fire‐prone landscapes are likely to be particularly sensitive because they often exhibit highly specialized pollination systems and provide no reward to pollinators, making them dependent on co‐flowering heterospecifics to attract and support pollinators. We investigated the hypothesis that fire‐driven changes in the local abundance of rewarding heterospecific flowers influence pollination in two rewardless Australian orchid species, Diuris maculata sensu lato and Caladenia tentaculata. Diuris maculata s.l. is thought to achieve pollination by mimicking papilionoid Fabaceae flowers. Caladenia tentaculata attracts male thynnine wasps through sexual deceit, and these wasps forage on the open‐access flowers of other taxa. We used a space‐for‐time substitution design with sites in different stages of post‐fire succession where we recorded capsule set in D. maculata s.l., pollinator visitation to C. tentaculata, the floral abundance of rewarding heterospecifics and abiotic conditions. Many rewarding taxa responded to fire age, but there was only weak evidence that capsule set in D. maculata s.l. was positively related to the local floral abundance of rewarding species. There was evidence of an overriding effect of rainfall on capsule set that may have obscured effects of the floral community. Visitation to C. tentaculata was not positively associated with any rewarding heterospecifics, and was negatively associated with rewarding Burchardia umbellata. Our preliminary findings highlight the need to account for multiple factors when trying to detect fire effects on pollination.

Convergent evolution of sexual deception via chromatic and achromatic contrast rather than colour mimicry

The Orchidaceae is characterised by the repeated evolution of sexual deception, one of the most specialised pollination strategies. In orchids, sexual deception involves long-range pollinator attraction via mimicry of female insect sex pheromones. At close range, visual signals involving colour mimicry, contrast to the background, and exploitation of pollinator sensory biases could attract pollinators, but remain largely untested. Here we focus on a remarkable system in which species from two only distantly related sexually deceptive orchid genera with strikingly different flowers (Drakaea livida and three species of Caladenia) share the same pollinator, males of the thynnine wasp Zaspilothynnus nigripes. We used spectral reflectance measurements and modelling to investigate pollinator perception of colour, including the first examination of overall colour patterns in flowers via colour pattern geometry analyses. Rather than closely matching the colours of female Z. nigripes, these orchids had strong chromatic and achromatic contrast against their backgrounds. For Caladenia, the sepals and petals show high contrast, while in D. livida, which has diminutive petals and sepals, it is the labellum that contrasts strongly against the background. Despite varying in colour, the Caladenia species all had strong within-flower contrast between a UV-bright central target (column and labellum) and a corolla of radiating stripes (petals and sepals). The colour pattern geometry analyses also indicated that the orchids’ overall colour patterns are highly conspicuous against their backgrounds. Contrast, UV, and target patterns could all enhance detection, and exploit pollinators’ innate preferences. Since colour contrast may function with a range of colours and floral forms, attracting pollinators via contrast rather than visual mimicry may be a critical but previously overlooked process facilitating the evolution of sexual deception.

Parapheromones for Thynnine Wasps

Sexually deceptive orchids produce floral volatiles that attract male insect pollinators. This interaction between flower and pollinator normally is highly specific. In the few cases where the chemical composition of the volatiles is known, the compounds have been found to be identical to those that comprise the sex pheromone of the female wasp. In this study, we investigated whether there is potential for flexibility in the molecular structure of the chemical cues used to mediate these specific interactions. Specifically, we asked whether strong sexual attraction can be maintained with structural modifications of sex pheromone components. Our study focused on the orchid, Drakaea glyptodon, which is pollinated by males of the thynnine wasp, Zaspilothynnus trilobatus. Three alkylpyrazines and a unique hydroxymethylpyrazine are components of the female produced sex pheromone of Z. trilobatus, and also the semiochemicals produced by the orchid that lures the males as pollinators. A blend of 2-butyl-3,5-dimethylpyrazine and 2-hydroxymethyl-3,5-diethyl-6-methylpyrazine (3:1) is as attractive as the full blend of four compounds. Therefore, in this study we substituted 2-hydroxymethyl-3,5-diethyl-6-methylpyrazine with one of five structurally related parapheromones in a blend with 2-butyl-3,5-dimethylpyrazine. All blends tested stimulated approaches by male wasps, with some also eliciting landing and attempted copulation. High-level calculations (G4(MP2)) showed the energy differences between the structural isomers were small, although the degree of sexual attraction varied, indicating the importance of structural factors for activity. One of the parapheromones, 2-hydroxymethyl-3,5-dimethyl-6-ethylpyrazine, elicited similar proportions of approaches, landings, and attempted copulations as the sex pheromone at the ratio and dose tested. The findings suggest that there is potential for chemical flexibility in the evolution of sexual deception.

Pollinator rarity as a threat to a plant with a specialized pollination system

An increasing diversity of highly specialized pollination systems are being discovered, many of which are likely to be vulnerable to anthropogenic landscape modification. Here, we investigate if a specialized pollination system limits the persistence of Caladenia huegelii (Orchidaceae), an endangered species pollinated by sexual deception of thynnine wasps. Once locally common in part of its geographical range, C. huegelii is now largely restricted to small habitat remnants in urban areas. Pollinator surveys coupled with DNA barcoding detected a single pollinator taxon, a small form of Macrothynnus insignis. Phylogenetic analysis revealed that small M. insignis from within the range of C. huegelii are strongly divergent from other wasp populations, suggesting that some reproductive isolation may exist. Although common in intact landscapes outside the range of C. huegelli, small M. insignis individuals were recorded at only 4% of sites in suitable C. huegelii habitat. Accordingly, reproductive success in C. huegelii was low compared with related Caladenia spp., with 33–60% of populations failing to set fruit in any given year. As such, populations are likely to now persist primarily through individual plant longevity rather than reproduction. Due to the low reproductive success of C. huegelii, ongoing human intervention will almost certainly be needed to sustain the species. Future research will need to focus on optimizing hand pollination to maintain reproduction and high seed fitness.

Absence of nectar resource partitioning in a community of parasitoid wasps

Parasitoid wasps occur in diverse communities, with the adults of most species sourcing carbohydrates from nectar or honeydew. However, the role of niche partitioning of nectar resources in maintaining diverse communities of parasitoid Hymenoptera is poorly known. To elucidate patterns of nectar resource use and test whether species partition resources, we investigated pollen loads in a community of parasitoid thynnine wasps in the biodiversity hotspot of southwestern Australia. In total, 304 thynnine wasps from 28 species were captured. Eighteen of these species are undescribed, highlighting the high diversity of unrecognized species in southwestern Australia. Pollen loads were detected on 111 individuals representing 19 species. Six pollen types were identified. All species that carried pollen primarily visited two tree species, Agonis flexuosa and Eucalyptus marginata, in the Myrtaceae. The other four pollen types were only recorded from single wasps. There was no evidence of nectar-resource partitioning. This may be due to these Myrtaceae producing abundant, open-faced flowers. Wasp species that were not recorded carrying pollen may utilise other carbohydrate sources, such as homopoteran honeydew. Niche partitioning is predicted to occur during the parasitoid larval phase of the life cycle. This study highlights the importance of nectariferous Myrtaceae in supporting diverse wasp communities. Further, two species of nectar-foraging wasps collected here are involved in the pollination of rare orchid species. Hence, conservation and management of habitats that support floriferous Myrtaceae are important for both the maintenance of diverse wasp communities, and the plants they pollinate.