Mycobiomes of two distinct clades of ambrosia gall midges (Diptera: Cecidomyiidae) are species-specific in larvae but similar in nutritive mycelia.

IMPORTANCE: Ambrosia gall midges are endophagous insect herbivores whose larvae live enclosed within a single gall for their entire development period. They may exhibit phytomycetophagy, a remarkable feeding mode that involves the consumption of plant biomass and mycelia of their cultivated gall symbionts. Thus, AGMs are ideal model organisms for studying the role of microorganisms in the evolution of host specificity in insects. However, compared to other fungus-farming insects, insect-fungus mutualism in AGMs has been neglected. Our study is the first to use DNA metabarcoding to characterize the complete mycobiome of the entire system of the gall-forming insects as we profiled gall surfaces, nutritive mycelia, and larvae. Interestingly, larval mycobiomes were significantly different from their nutritive mycelia, although Botryosphaeria dothidea dominated the nutritive mycelia, regardless of the evolutionary separation of the tribes studied. Therefore, we confirmed a long-time hypothesized paradigm for the important evolutionary association of this fungus with AGMs.

New Cladosporium Species from Normal and Galled Flowers of Lamiaceae.

A series of isolates of Cladosporium spp. were recovered in the course of a cooperative study on galls formed by midges of the genus Asphondylia (Diptera, Cecidomyidae) on several species of Lamiaceae. The finding of these fungi in both normal and galled flowers was taken as an indication that they do not have a definite relationship with the midges. Moreover, identification based on DNA sequencing showed that these isolates are taxonomically heterogeneous and belong to several species which are classified in two different species complexes. Two new species, Cladosporium polonicum and Cladosporium neapolitanum, were characterized within the Cladosporium cladosporioides species complex based on strains from Poland and Italy, respectively. Evidence concerning the possible existence of additional taxa within the collective species C. cladosporioides and C. pseudocladosporioides is discussed.

Delineating the Asphondylia prosopidis Complex (Diptera: Cecidomyiidae): Possible Biological Control Agents of Honey Mesquite.

The Asphondylia prosopidis Cockrell complex with its fungal associate produces four distinct bud galls on honey mesquite (Prosopis glandulosa). They are considered as potential biological control agents to reduce seed production of invasive mesquites. Previous studies suggest that the complex may consist of more than one gall midge species or biotype. Therefore, before conducting host specificity tests, it is essential to understand the relationships among the gall midges in the complex. Each gall type was collected from sympatric regions in Arizona, New Mexico, and Texas for four years. Here we show that midges producing each gall type were clearly separated based on phylogenetic analysis using DNA sequences in the cytochrome oxidase subunit I region. Furthermore, we confirmed that morphological differences between pupae from each gall type were discernible, although variable, using a scanning electron microscope. Based on these differences, we suggest that the A. prosopidis complex consists of four different gall midge species, three of which are cryptic species. Among them, Asphondylia species producing a barrel gall type and A. prosopidis producing the original gall type are potential biological control agents of P. glandulosa because they are multivoltine species with four to five generations per year that complete their annual life cycle exclusively on flower buds of mesquite.

Polyploidy in creosote bush (Larrea tridentata) shapes the biogeography of specialist herbivores.

AIM: Whole-genome duplication (polyploidy) can influence the biogeography and ecology of plants that differ in ploidy level (cytotype). Here, we address how two consequences of plant polyploidy (parapatry of cytotypes and altered species interactions) shape the biogeography of herbivorous insects. LOCATION: Warm deserts of North America. TAXA: Gall midges (Asphondylia auripila group, Diptera: Cecidomyiidae) that attack three parapatric cytotypes of creosote bush (Larrea tridentata, Zygophyllaceae). METHODS: We surveyed Asphondylia species diversity at 177 sites across a 2300-km extent. After noting a correspondence between the distributions of eight Asphondylia species and L. tridentata cytotypes, we fine-mapped Asphondylia species range limits with transects spanning cytotype contact zones. We then tested whether plant-insect interactions and/or abiotic factors explain this coincidence by (1) comparing attack rates and gall midge communities on alternative cytotypes in a narrow zone of sympatry and (2) using species distribution models (SDMs) to determine if climatically suitable habitat for each midge species extended beyond cytotype contact zones. RESULTS: The range limits of 6/17 Asphondylia species (including two novel putative species confirmed with COI sequencing) perfectly coincided with the contact zone of diploid and tetraploid L. tridentata. One midge species was restricted to diploid host plants while five were restricted to tetraploid and hexaploid host plants. Where diploid and tetraploid L. tridentata are sympatric, cytotype-restricted midge species more frequently attacked their typical host and Asphondylia community structure differed markedly between cytotypes. SDMs predicted that distributions of cytotype-restricted midge species were not constrained by climatic conditions near cytotype contact zones. MAIN CONCLUSIONS: Contact zones between plant cytotypes are dispersal barriers for many Asphondylia species due to plant-insect interactions. The distribution of L. tridentata cytotypes therefore shapes herbivore species ranges and herbivore community structure across North American deserts. Our results demonstrate that polyploidy in plants can affect the biogeography of ecological communities.

Asphondylia fructicola, a new species of Cecidomyiidae (Diptera) associated with Solanum sp. (Solanaceae) from Brazil / Asphondylia fructicola, uma nova espécie de Cecidomyiidae (Diptera) associada com Solanum sp. (Solanaceae) do Brasil

Asphondylia fructicola sp. nov. is described and illustrated on the basis of the larva, pupa, male, female, and gall. This species induces galls on fruits of Solanum sp. (Solanaceae) in Amazonia, Brazil.

Asphondylia fructicola sp. nov. é descrita e ilustrada com base na larva, pupa, macho, fêmea e galha. Essa espécie induz galhas nos frutos de Solanum sp. (Solanaceae) na Amazônia, Brasil.