Abstract
The brown marmorated stink bug (Halyomorpha halys) has emerged as one of the most important invasive insect pests in the United States. Functional genomics in H. halys remains unexplored as molecular resources in this insect have recently been developed. To facilitate functional genomics research, we evaluated ten common insect housekeeping genes (RPS26, EF1A, FAU, UBE4A, ARL2, ARP8, GUS, TBP, TIF6 and RPL9) for their stability across various treatments in H. halys. Our treatments included two biotic factors (tissues and developmental stages) and two stress treatments (RNAi injection and starvation). Reference gene stability was determined using three software algorithms (geNorm, NormFinder, BestKeeper) and a web-based tool (RefFinder). The qRT-PCR results indicated ARP8 and UBE4A exhibit the most stable expression across tissues and developmental stages, ARL2 and FAU for dsRNA treatment and TBP and UBE4A for starvation treatment. Following the dsRNA treatment, all genes except GUS showed relatively stable expression. To demonstrate the utility of validated reference genes in accurate gene expression analysis and to explore gene silencing in H. halys, we performed RNAi by administering dsRNA of target gene (catalase) through microinjection. A successful RNAi response with over 90% reduction in expression of target gene was observed.
Highlights
Determining gene function within a genome provides insight into the molecular interactions between organisms and their environments
We demonstrated the utility of validated reference genes in accurate expression analysis during the exploration for successful gene silencing in H. halys by dsRNA injection
Optimization of qRT-PCR assay for candidate genes
Summary
Determining gene function within a genome (i.e. functional genomics) provides insight into the molecular interactions between organisms and their environments. Insects present good test cases for functional genomics because of their fast generation time, reproductive output, ease of rearing large populations, and a wealth of genomic knowledge. Two techniques are PLOS ONE | DOI:10.1371/journal.pone.0152730. Functional Genomics in Stink Bug following interests. There are no patents, products in development or marketed products to declare. This does not alter the authors’ adherence to all the PLOS ONE policies on sharing data and materials, as detailed on-line in the guide for authors
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