BACKGROUND: The phlebotomine sand fly Phlebotomus perniciosus (Diptera: Psychodidae, Phlebotominae) is a major Old World vector of the protozoan Leishmania infantum, the etiological agent of visceral and cutaneous leishmaniases in humans and dogs, a worldwide re-emerging diseases of great public health concern, affecting 101 countries. Despite the growing interest in the study of this sand fly species in the last years, the development of genomic resources has been limited so far. To increase the available sequence data for P. perniciosus and to start studying the molecular basis of the sexual differentiation in sand flies, we performed whole transcriptome Illumina RNA sequencing (RNA-seq) of adult males and females and de novo transcriptome assembly. RESULTS: We assembled 55,393 high quality transcripts, of which 29,292 were unique, starting from adult whole body male and female pools. 11,736 transcripts had at least one functional annotation, including full-length low abundance salivary transcripts, 981 transcripts were classified as putative long non-coding RNAs and 244 transcripts encoded for putative novel proteins specific of the Phlebotominae sub-family. Differential expression analysis identified 8590 transcripts significantly biased between sexes. Among them, some show relaxation of selective constraints when compared to their orthologs of the New World sand fly species Lutzomyia longipalpis. CONCLUSIONS: In this paper, we present a comprehensive transcriptome resource for the sand fly species P. perniciosus built from short-read RNA-seq and we provide insights into sex-specific gene expression at adult stage. Our analysis represents a first step towards the identification of sex-specific genes and pathways and a foundation for forthcoming investigations into this important vector species, including the study of the evolution of sex-biased genes and of the sexual differentiation in phlebotomine sand flies.

Department of Biology University of Naples Federico 2 Naples Italy

Department of Infectious Parasitic and Immunomediated Diseases Istituto Superiore di Sanità Rome Italy

Department of Parasitology Charles University Prague Czech Republic

Genomics Core Facility EMBL Heidelberg Germany

Institute for High Performance Computing and Networking ICAR CNR Naples Italy

Institute of Molecular Life Science University of Zurich Zurich Switzerland SIB Swiss Institute of Bioinformatics University of Zurich Zurich Switzerland

National Research Council Institute of Genetics and Biophysics Naples Italy

Stazione Zoologica Anton Dohrn Naples Italy

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$a De novo assembly and sex-specific transcriptome profiling in the sand fly Phlebotomus perniciosus (Diptera, Phlebotominae), a major Old World vector of Leishmania infantum / $c V. Petrella, S. Aceto, F. Musacchia, V. Colonna, M. Robinson, V. Benes, G. Cicotti, G. Bongiorno, L. Gradoni, P. Volf, M. Salvemini,

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$a BACKGROUND: The phlebotomine sand fly Phlebotomus perniciosus (Diptera: Psychodidae, Phlebotominae) is a major Old World vector of the protozoan Leishmania infantum, the etiological agent of visceral and cutaneous leishmaniases in humans and dogs, a worldwide re-emerging diseases of great public health concern, affecting 101 countries. Despite the growing interest in the study of this sand fly species in the last years, the development of genomic resources has been limited so far. To increase the available sequence data for P. perniciosus and to start studying the molecular basis of the sexual differentiation in sand flies, we performed whole transcriptome Illumina RNA sequencing (RNA-seq) of adult males and females and de novo transcriptome assembly. RESULTS: We assembled 55,393 high quality transcripts, of which 29,292 were unique, starting from adult whole body male and female pools. 11,736 transcripts had at least one functional annotation, including full-length low abundance salivary transcripts, 981 transcripts were classified as putative long non-coding RNAs and 244 transcripts encoded for putative novel proteins specific of the Phlebotominae sub-family. Differential expression analysis identified 8590 transcripts significantly biased between sexes. Among them, some show relaxation of selective constraints when compared to their orthologs of the New World sand fly species Lutzomyia longipalpis. CONCLUSIONS: In this paper, we present a comprehensive transcriptome resource for the sand fly species P. perniciosus built from short-read RNA-seq and we provide insights into sex-specific gene expression at adult stage. Our analysis represents a first step towards the identification of sex-specific genes and pathways and a foundation for forthcoming investigations into this important vector species, including the study of the evolution of sex-biased genes and of the sexual differentiation in phlebotomine sand flies.

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$a Musacchia, F $u Stazione Zoologica "Anton Dohrn", Naples, Italy.

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$a Colonna, V $u National Research Council, Institute of Genetics and Biophysics, Naples, Italy.

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$a Robinson, M $u Institute of Molecular Life Science, University of Zurich, Zurich, Switzerland. SIB-Swiss Institute of Bioinformatics, University of Zurich, Zurich, Switzerland.

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$a Benes, V $u Genomics Core Facility, EMBL, Heidelberg, Germany.

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$a Cicotti, G $u Institute for High Performance Computing and Networking, ICAR-CNR, Naples, Italy.

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$a Bongiorno, G $u Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy.

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$a Gradoni, L $u Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy.

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$a Volf, P $u Department of Parasitology, Charles University, Prague, Czech Republic.

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$a Salvemini, M $u Department of Biology, University of Naples Federico II, Naples, Italy. marco.salvemini@unina.it.

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