Tolerances of Apple Maggot (Diptera: Tephritidae) Larvae and Different Age Puparia to Water Flotation and Immersion

Tolerance of terrestrial insects in temperate regions to water immersion and hypoxia has rarely been studied but can be an important adaptation to moist environments, with implications for insect dispersal through waterways. In the Pacific Northwest of the United States, apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), can be found in riparian habitats subject to flooding. Here, survival of R. pomonella larvae and different age puparia after flotation or immersion in 13.3°C or 21.1°C water for 1–12 d was determined. Larvae sank in water and when submerged for 1 or 2 d suffered greater mortality than control larvae. Fewer young (1–2 d old) than older puparia (13–15 d old) floated in water. When immersed in water for 1–12 d, young puparia suffered greater mortality than older puparia, which were not affected by water immersion. Consequently, fewer adult flies eclosed from puparia that had been water treated when young than older. Adult flies from pre-chill and post-chill puparia that had been water treated eclosed later than control flies, but treatment flies survived about 60 d and reproduced. Although newly-formed puparia are susceptible to hypoxic water conditions, increased buoyancy and water tolerance occur rapidly after formation, perhaps making survival possible and allowing water-borne dispersal of older puparia.

The Eclosion of Rhagoletis pomonella (Diptera: Tephritidae) Under Different Chill Durations and Simulated Temperate and Tropical Conditions

The apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), is a serious pest of apple in North America that is subject to quarantine measures to prevent its spread to currently pest-free regions, including the tropics. How the fly may survive in warmer climates is unclear. Here, we studied the effects of exposing postchill puparia to simulated temperate and tropical environmental conditions on eclosion of R. pomonella from Washington State, U.S.A. Puparia were chilled for 0–30 wk at 3°C and then held under four postchill conditions: A = 23°C, 16:8 L:D, 40% RH; B = 26°C, 12:12 L:D, 80% RH; C = 26°C, 16:8 L:D, 80% RH; and D = 23°C, 12:12 L:D, 40% RH, with B and D representing tropical conditions and A and C temperate conditions. Within each chill duration, total numbers of flies eclosed were equally high in tropical treatment B and temperate treatment C, while they were lower in treatments A and D. Mean weeks of the first eclosion in treatments B and C were earlier than in treatment D; mean week of peak eclosion and 50% eclosion in treatments A, B, and C were earlier than in treatment D. Eclosion spans in treatments A, B, and D were generally shorter than in treatment C. Results suggest that if introduced into a humid tropical country, R. pomonella puparia from Washington State could produce adult flies, regardless of chill duration or lack of chilling during the pupal stage, but whether flies could establish there would require further study.

Identifying Diagnostic Genetic Markers for a Cryptic Invasive Agricultural Pest: A Test Case Using the Apple Maggot Fly (Diptera: Tephritidae)

Insect pests destroy ~15% of all U.S. crops, resulting in losses of $15 billion annually. Thus, developing cheap, quick, and reliable methods for detecting harmful species is critical to curtail insect damage and lessen economic impact. The apple maggot fly, Rhagoletis pomonella, is a major invasive pest threatening the multibillion-dollar apple industry in the Pacific Northwest United States. The fly is also sympatric with a benign but morphologically similar and genetically closely related species, R. zephyria, which attacks noncommercial snowberry. Unambiguous species identification is essential due to a zero-infestation policy of apple maggot for fruit export. Mistaking R. zephyria for R. pomonella triggers unnecessary and costly quarantines, diverting valuable control resources. Here we develop and apply a relatively simple and cost-effective diagnostic approach using Illumina sequencing of double-digest restriction site-associated DNA markers. We identified five informative single-nucleotide polymorphisms (SNPs) and designed a diagnostic test based on agarose gel electrophoresis of restriction enzyme-digested polymerase chain reaction amplification products (RFLPs) to distinguish fly species. We demonstrated the utility of this approach for immediate, 1-d species identification by scoring apple- and snowberry-infesting flies from known hosts, reared from fruit collected at 11 sites throughout Washington. However, if immediate diagnosis is not required, or hundreds to thousands of specimens must be assessed, then a direct Illumina-based sequencing strategy, similar to that used here for diagnostic SNP identification, can be powerful and cost-effective. The genomic strategy we present is effective for R. pomonella and also transferable to many cryptic pests.