Abstract
Halyomorpha halys (Stål) (Heteroptera: Pentatomidae), the brown marmorated stink bug, is an invasive agricultural and nuisance pest rapidly expanding its incidence in North America. This voracious pest poses a significant threat to rural and urban agriculture, especially to specialty crops such as apples, grapes and ornamentals, as well as staple crops including soybean and corn. The object of this study was to generate transcript sequence resources for H. halys. RNA-seq libraries derived from distinct developmental stages and sexes were sequenced and assembled into 248,569 putatively unique transcripts (PUTs). PUTs were segmented into three disjoint tiers of varying reliability, with 4,794 classified as gold tier (highest quality), 16,878 as silver, and 14,357 as bronze. The gold-tier PUTs associated with 2,580 distinct non-redundant protein sequences from the NCBI NR database—1,785 of these (69%) mapped to annotated UniProtKB database proteins, from which 1,273 unique Pfam families and 459 unique Molecular Function GO terms were encountered. Of the silver tier's 6,527 PUTs associated with unique proteins, 4,193 mapped to UniProtKB (64%), from which 1,941 and 640 unique Pfam and Molecular Function GO terms were extracted. H. halys PUTs related to important life processes like immunity, endocrinology, reproduction, development, behavior, neurotransmission, neurotoxicity, olfaction, and small RNA pathways were validated through quantitative Real-Time PCR (qRT-PCR) for differential expression during distinct life stages (eggs, 2nd instar nymphs, 4th instar nymphs, female adults, male adults). PUTs similar to hypothetical proteins identified in symbiont microbes, including Pantoea and Nosema species, were more abundantly expressed in adults versus nymphs. These comprehensive H. halys transcriptomic resources can be utilized to aid development of novel control methodologies to disrupt life processes; to conduct reverse genetic screens to determine host gene function; and to design environmentally unobtrusive means to control host populations or target specific H. halys life stages, such as molecular biopesticides.
Highlights
Halyomorpha halys (Stal) (Heteroptera: Pentatomidae), the brown marmorated stink bug (BMSB), is an invasive insect native to Asia (China, Taiwan, Korea, and Japan) that has emerged in the last decade as an important major insect pest in the United States, Canada and Europe and has become the top invasive insect research priority for the USDA Agricultural Research Service
Qualitative analysis of H. halys RNA-Seq data Figure 1 shows the RNA-Seq assembly and analysis pipeline used in this study, which is further detailed in Materials and Methods
The 439,615,225 RNA-Seq reads surviving quality control procedures were assembled into a total of 248,569 putatively unique transcripts (PUTs)
Summary
Halyomorpha halys (Stal) (Heteroptera: Pentatomidae), the brown marmorated stink bug (BMSB), is an invasive insect native to Asia (China, Taiwan, Korea, and Japan) that has emerged in the last decade as an important major insect pest in the United States, Canada and Europe and has become the top invasive insect research priority for the USDA Agricultural Research Service. Numerous H. halys differentially expressed transcripts were associated with functions in critical life processes, including immune response, endocrinology, reproduction, growth, development, behavior, neurotransmission, neurotoxicity, olfaction, detoxification, insecticide resistance, and other categories.
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