Sample collection, sequencing and mtgenome assembly
All samples were collected in
China (Table S1), and then stored in 95% alcohol at -20 °C until DNA
extraction. These
samples
were identified in morphology, 19 species were selected for sequencing,
and their identification were confirmed by subsequent COX1comparison with BOLD (http://boldsystems.org/index.php) (Hebert et
al ., 2003). The total genomic DNA was extracted from the thorax and leg
muscle tissue by DNeasy Blood and
Tissue kit (Qiagen, Duesseldorf, Germany) according to the instructions
of manufacturer. Concentration of extracted genomic DNA was determined
by Qubit 2.0 (Invitrogen, Shanghai, China). The 350 bp small fragment
libraries were constructed, and then sequenced using the Illumina Hiseq
2500 (San Diego, CA) with 150 bp paired-end reads in
Shenzhen Huitong Biotechnology Co.
Ltd (Shenzhen, China). After removing the adapters, and unpaired, short
and low qualitied reads, clean reads from mtgenomes were extracted using
a BLAST (Altschul et al. , 1990) search against known
Tenebrionoidea mtgenome sequences, and then used for de novomtgenome assembly with SPAdes v. 3.9.0 (Bankevich et al. , 2012).
The contigs of mtgenome were extracted and assembled into mtgenomes
through searching against the reference sequences using PRICE
(paired-read iterative contig extension) by NOVOPlasty version 2.6.2
(Dierckxsens et al. , 2016).