The genus
Tipula
Linnaeus,
1758
is a large group of crane flies with more than 2400 known species from 41 subgenera. In this study, we report the first complete mitochondrial (mt) genome sequence of the subgenus
Tipula (Yamatotipula)
, which is a circular molecule of 15,668 bp with an AT content of 77.2%. The mt genome contains 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and a long non-coding region. Three conserved overlapping regions, 8 bp between
tRNA
Trp
and
tRNA
Cys
, 7 bp between
ATP8
and
ATP6
, and 7 bp between
ND4
and
ND4L
, are found. Phylogenetic analysis reveals that the Tipulomorpha includes the family Trichoceridae and the Trichoceridae is sister-group to the remaining Tipulomorpha.
In this study, we describe a cell line, Ms-10C, cloned from the line QAU-Ms-E-10 (simplified Ms-10), an embryonic line from Mythimna separata. The cloned cell line was significantly more sensitive to nucleopolyhedrovirus (NPV). Ms-10C cells were mainly spherical with a diameter of 14.42 ± 2.23 μm. DNA amplification fingerprinting (DAF) confirmed the profile of PCR-amplified bands of the cloned cell line was consistent with those of the parental cell line, Ms-10. The sequencing result of the mitochondrial cytochrome c oxidase I (mtCO I) fragment confirmed that the amplified 636-bps mtCOI fragment was 100% identical to that of M. separata. Its chromosomes exhibited the typical characters of lepidopteran cell lines. Its population doubling time was 42.2 h at 27°C. Ms-10C was more sensitive than Ms-10 to both Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and M. separata nucleopolyhedrovirus (MsNPV). At 4 d post infection, the infection rates of two viruses reached 94.2 and 92.3%, respectively. The availability of this cell clone strain will provide a useful tool for the basic research on nucleopolyhedrovirus and for potential application in expression of recombinant proteins with baculovirus expression vector system.
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