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Mitochondrial DNA, Early Online: 1–3

! 2015 Informa UK Ltd. DOI: 10.3109/19401736.2015.1022748

MITOGENOME ANNOUNCEMENT

Complete mitochondrial genome of Lutzomyia (Nyssomyia) umbratilis

(Diptera: Psychodidae), the main vector of Leishmania guyanensis
Arthur Kocher1, Jean-Charles Gantier2, Hélène Holota1, Céline Jeziorski3,4, Eric Coissac5, Anne-Laure Bañuls6,
Romain Girod7, Pascal Gaborit7, and Jérôme Murienne1
1
CNRS, Université Toulouse III Paul Sabatier, ENFA; UMR5174 EDB (Laboratoire Evolution et Diversité Biologique), Toulouse, France, 2Laboratoire des
Identifications Fongiques et Entomo-parasitologiques, Mennecy, France, 3INRA, UAR 1209 Département de génétique animale, INRA Auzeville,
Castanet-Tolosan, France, 4GeT–PlaGe, Genotoul, INRA Auzeville, Castanet-Tolosan, France, 5Laboratoire d’Ecologie Alpine, CNRS UMR 5553,
Université Joseph Fourier, Grenoble, France, 6UMR MIVEGEC (IRD 224 – CNRS 5290), Université de Montpellier), Montpellier, France, and
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7
Medical Entomology Unit, Institut Pasteur de la Guyane, Cayenne Cedex, French Guiana

Abstract Keywords
The nearly complete mitochondrial genome of Lutzomyia umbratilis Ward & Fraiha, 1977 Genome skimming, illumina Hiseq,
(Psychodidae: Phlebotominae), considered as the main vector of Leishmania guyanensis, is leishmaniasis, next-generation sequencing,
presented. The sequencing has been performed on an Illumina Hiseq 2500 platform, with a Sandfly, shotgun sequencing
genome skimming strategy. The full nuclear ribosomal RNA segment was also assembled. The
mitogenome of L. umbratilis was determined to be at least 15,717 bp-long and presents an History
architecture found in many mitogenomes of insect (13 protein-coding genes, 22 transfer RNAs,
Received 5 February 2015
For personal use only.

two ribosomal RNAs, and one non-coding region also referred as the control region). The
control region contains a large repeated element of c. 370 bp and a poly-AT region of unknown Accepted 15 February 2015
length. This is the first mitogenome of Psychodidae to be described. Published online 22 May 2015

Lutzomyia sensu lato is a New World genus of sandflies
(Psychodidae: Phlebotominae) that currently includes 477
described species. It comprises all the proven vector species of
Leishmaniasis in the New World (Killick-Kendrick, 1990).
Lutzomyia umbratilis (Ward & Fraiha 1977) is distributed in
several countries of northern South America (Scarpassa &
Alencar, 2012) and is considered to be the main vector of
Leishmania guyanensis (Pajot et al., 1986). Here, the complete
mitochondrial genome of L. umbratilis was sequenced using a
genome-skimming approach (Cally et al., 2014; Kocher et al.,
2014, 2015). This is the first mitogenome of Psychodidae to be
described.
Specimens of L. umbratilis were collected using CDC light
traps in the surrounding of Saint-Georges de l’Oyapock, French
Guiana on 2 October 2007, processed in the Pasteur institute in
Cayenne and stored at the LIFE laboratory, Mennecy, France.
After traditional morphological identification, heads, wings, and
genitalia were kept as vouchers (IDs: 1664B, 1667B, and 1669B).
Total genomic DNA was extracted from the thoraxes using the
DNeasy Blood and Tissue kit (Qiagen, Valencia, CA). DNA was
then sent for library construction and sequencing to the GeT-
PlaGe core facilities of Genotoul (Toulouse, France). The library
was constructed with 162 ng of DNA using the Illumina TruSeq
Nano DNA Sample Prep Kit (Illumina Inc. San Diego, CA). The
library was hybridized on 1/24th of a lane of Illumina Hiseq 2500
flow cell using the Illumina TruSeq PE Cluster Kit v.3 (Illumina
Correspondence: Dr Arthur Kocher, CNRS, Toulouse, France. E-mail:
arthur.kocher@gmail.com
Inc., San Diego, CA). Paired-end reads of 100 nucleotides were
collected using the Illumina TruSeq SBS Kit v.3 (Illumina Inc.,
San Diego, CA) (200 cycles). The mitochondrial genome and the
nuclear ribosomal gene segment were assembled using a genome
walking approach and annotated as previously described (Kocher
et al., 2014). The sequences were deposited in GenBank under the
accession numbers KP702938 and KP702939. Because this is the
first mitogenome of Psychodidae available, we validated our
sequence based on the 50 region of the mitochondrial gene COI
using the Barcoding of Life Database Identification Engine
(http://www.boldsystems.org, Figure 1).
These results demonstrate that for a relatively small cost
(below 280 euros), it is possible to efficiently assemble a
complete sandfly mitogenome with sufficient coverage (the mean
coverage in the coding region was 84  with a minimum of 30)
as well as a complete nuclear ribosomal cluster (the mean
coverage was 101  with a minimum of 31). The complete
mitogenome of L. umbratilis is determined to be at least 15,717 bp
long and presents an organization shared with many insects
(Boore, 1999). The typical set of 37 genes (13 protein-coding
genes, 22 transfer RNA genes, and two ribosomal RNA genes)
and one control region was identified. All protein-coding genes
started with an ATN codon. Nine TAA and one TAG stop codons
were identified. Three incomplete stop codons (TA or T) were
found adjacent to transfer RNAs encoded on the same strand,
consistent with the punctuation model for primary transcripts
processing followed by 30 polyadenylation of mature mRNA
(Nagaike, 2005; Ojala et al., 1981). The sequence of the control
region was not completely assembled, but was determined to be at
least 882 bp long. It contained at least two almost identical
 2 A. Kocher et al.
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For personal use only.
Figure 1. Neighbor-joining tree provided by the BOLD Identification Engine for the validation of our sample. The tree is based on the 50 region of the
mitochondrial gene COI, which is the typical DNA barcode for animals. Our sequence is annotated with its GenBank accession number.
tandem-repeats of a ca 370 bp-long element and a poly-AT region
of unknown length. The latter is probably related with a very low
sequencing coverage of the region and the inability to complete
the assembly.
Acknowledgements
We would like to thank Romuald Carinci and Jean Issaly for their
involvement in the collection and processing of the samples.
Declaration of interest
This work was supported by ‘‘Investissement d’Avenir’’ grants managed
by Agence Nationale de la Recherche (CEBA, ANR-10-LABX-25-01;
TULIP, ANR-10-LABX-41, and ANR-11-IDEX-0002-02) as well as
Mitochondrial DNA, Early Online: 1–3
project METABAR (ANR-11-BSV7-0020). The collection of sandflies
was performed in the framework of the French National Project ANR 06-
SEST-20 IAEL. The authors report that they have no conflicts of interest.
The authors alone are responsible for the content and writing of the paper.
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Complete mitogenome of L. (Nyssomyia) umbratilis 3
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Supplementary material: Genitalia of male Lutzomyia umbratilis (voucher’s ID: 1669B)