Genome-wide analyses of ATP-Binding Cassette (ABC) transporter gene family and its expression profile related to deltamethrin tolerance in non-biting midge Propsilocerus akamusi.

Non-biting midges are dominant species in aquatic systems and often used for studying the toxicological researches of insecticides. ATP-binding cassette (ABC) transporters represent the largest known members in detoxification genes but is little known about their function in non-biting midges. Here, we selected Propsilocerus akamusi, widespread in urban streams, to first uncover the gene structure, location, characteristics, and phylogenetics of chironomid ABC transporters at genome-scale. Fifty-seven ABC transporter genes are located on four chromosomes, including eight subfamilies (ABCA-H). The ABCC, ABCG, and ABCH subfamilies experienced the duplication events to different degrees. The study showed that expression of the PaABCG17 gene is uniquely significantly elevated, with deltamethrin concentration increasing (1, 4, and 20 ug/L) both in RNA-seq and qPCR results. Additionally, the ABC transporter members of other six chironomids with assembled genomes are first described and used to investigate the characteristic of those living in the different adverse habitats. The ABC transporter frame for Propsilocerus akamusi and its transcriptomic results lay an important foundation for providing valuable resources for understanding the ABC transporter function in insecticide toxification of this species as well as those of other non-biting midges. The PaABCG17 gene is shown to play an important role in deltamethrin detoxification, and it functions need to be further investigated and might be used in the management of insecticide-resistance in chironomid adults.

A chromosome level genome assembly of Propsilocerus akamusi to understand its response to heavy metal exposure.

Chironomidae species are universally used for studying the impact of pollutants in aquatic systems. The nonbiting midge Propsilocerus akamusi is often found in urban streams and is suitable for use as a toxicological bioindicator. However, few studies have previously examined metal stress in this species. We sequenced the genome of this urban midge to address this question. Here, we present the first chromosome-level genome of P. akamusi, obtained from Illumina short-read and PacBio long-read sequences with Hi-C technology. The size of the very small assembled genome was 85.83 Mb with a contig N50 of 6.2 Mb and a scaffold N50 of 26.1 Mb. This assembly revealed significant expansion of haemoglobin (Hb) genes, some of which formed large tandem repeats. Transcriptomic studies for copper tolerance identified four genes in the tandem array that were highly expressed, all of which presented intron loss. This characteristic might highlight the potential role of Hb genes in copper tolerance. Additionally, detoxification genes, chemosensory genes and heat shock protein genes of this midge were identified, some of which are associated with metal stress. The high-quality assembled genome of P. akamusi and the transcriptomic analyses provide new insight into the molecular mechanisms of heavy metal stress. Our comparison of the P. akamusi genome with other dipteran genomes provides valuable resources for understanding the evolutionary history, genetics, and ecology of this species as well as those of other midges.

Identification and expression pattern of chemosensory genes in the transcriptome of Propsilocerus akamusi.

Chironomidae is the most ecologically diverse insects in aquatic and semi-aquatic habitats. Propsilocerus akamusi (Tokunaga) is a dominant and ubiquitous chironomid species in Eastern Asia and its morphologically unique larvae are also considered as indicator organisms to detect water contamination, potential toxicity and waterborne pathogens. Since few studies to date have focused on the olfactory system of P. akamusi, our study aims to elucidate the potential functions of chemosensory genes in P. akamusi. In our study, we found that although signals released from male groups might attract female swarmers, it was a completely male-dominated mating process. Sequencing the transcriptome of P. akamusi on an Illumina HiSeq platform generated 4.42, 4.46 and 4.53 Gb of clean reads for heads, legs, and antennae, respectively. 27,609 unigenes, 20,379 coding sequences (CDSs), and 8,073 simple sequence repeats were finally obtained. The gene-level differential expression analysis demonstrated variants among three different tissues, including 2,019 genes specifically expressed in heads, 1,540 genes in legs, and 2,071 genes in antennae. Additionally, we identified an assortment of putative olfactory genes consisting of 34 odorant binding proteins, 17 odorant receptors, 32 gustatory receptors, 22 ionotropic receptors, six chemosensory proteins as well as 3 sensory neuron membrane proteins; their relative abundances in the above three tissues were also determined by RT-qPCR. Our finding could allow a more plausible understanding of certain olfaction-mediated behaviors in groups of this macroinvertebrate.