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The Complete Mitochondrial Genome of Trabala Vishnou Guttata (Lepidoptera: Lasiocampidae) and the Related Phylogenetic Analyses

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The Complete Mitochondrial Genome of Trabala Vishnou Guttata (Lepidoptera: Lasiocampidae) and the Related Phylogenetic Analyses The complete mitochondrial genome of Trabala vishnou guttata (Lepidoptera: Lasiocampidae) and the related phylogenetic analyses Liuyu Wu, Xiao Xiong, Xuming Wang, Tianrong Xin, Jing Wang, Zhiwen Zou & Bin Xia Genetica An International Journal of Genetics and Evolution ISSN 0016-6707 Volume 144 Number 6 Genetica (2016) 144:675-688 DOI 10.1007/s10709-016-9934-x 1 23 Your article is protected by copyright and all rights are held exclusively by Springer International Publishing Switzerland. This e- offprint is for personal use only and shall not be self-archived in electronic repositories. If you wish to self-archive your article, please use the accepted manuscript version for posting on your own website. 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The link must be accompanied by the following text: "The final publication is available at link.springer.com”. 1 23 Author's personal copy Genetica (2016) 144:675–688 DOI 10.1007/s10709-016-9934-x The complete mitochondrial genome of Trabala vishnou guttata (Lepidoptera: Lasiocampidae) and the related phylogenetic analyses 1 1 2 1 1 Liuyu Wu • Xiao Xiong • Xuming Wang • Tianrong Xin • Jing Wang • 1 1 Zhiwen Zou • Bin Xia Received: 20 May 2016 / Accepted: 17 October 2016 / Published online: 21 October 2016 Ó Springer International Publishing Switzerland 2016 Abstract The bluish yellow lappet moth, Trabala vishnou related species (Dendrolimus taxa) are clustered on Lasio- guttata is an extraordinarily important pest in China. The campidae group. It is a sister clade relationship between complete mitochondrial genome is sequenced and deter- Lasiocampidae and other families in Bombycoidea with a mined firstly, which is based on traditional PCR amplifica- bootstrap value of 83 % and a posterior probability of 0.75. tion and primer walking methods with a length of 15,281 bp, This study supports that Lasiocampidae may be independent including 13 protein-coding (PCG) genes, 22 transfer RNA from Bombycoidea. (rRNA) genes, two ribosomal RNA (tRNA) genes, and an A ? T-rich region. The gene order and orientation of the T. Keywords Trabala vishnou guttata Á Lasiocampidae Á vishnou guttata mitogenome were identical to the other Mitochondrial genome Á Phylogenetic analyses sequenced Lasiocampidae species. The overall nucleotide composition of T. vishnou guttata is A (40.27 %), T (40.59 %), C (11.58 %) and G (7.56 %), respectively. All Introduction the PCGs initiate with the three orthodox start codons ATN except for coxI with CGA start codon. Three PCGs (coxI, Trabala vishnou guttata (Matsumura, 1990) (Lepidoptera: coxII and nad4) used incomplete stop codon T, while the Lasiocampidae) is a sort of widely pest damaging many other 10 PCGs terminate with complete stop codon TAA. All fruit trees with an extensively distribution in south of tRNA genes have a typical clover-leaf structure except for China, for example Taiwan, and it has been identified the absence of a dihydrouridine arm in trnS (AGN). The belong to Lasiocampidae family in the superfamily of length of A ? T-rich region is 383 bp. Phylogeny is estab- Bombycoidea (Liu and Wu 2006). It has been investigated lished to reveal the genetic relationship between T. vishnou by the author that the larva, as a kind of local dominant guttata and other lepidopteran species based on 13 PCGs species feed on many plants leaves which has a serious nucleotide sequences of the sequenced species (32 taxa) by influence on the ecological constructions and economic Maximum likelihood and Bayesian methods. Phylogenetic development immediately. Otherwise, it is reported that analyses presents that T. vishnou guttata and its closely toxic hair of the similar lepidopteran larva is very easy to penetrate into the skin causing swelling and pain during field investigations and studies (Han et al. 2005). Although some studies have concentrated on its ecological and bio- Electronic supplementary material The online version of this article (doi:10.1007/s10709-016-9934-x) contains supplementary logical characteristics for chemical control and biological material, which is available to authorized users. control, its genetic researches are rarely reported. Phylo- genetic relationships of T. vishnou guttata have not been & Bin Xia investigated previously. There only has been a part of coxI [email protected] gene published in NCBI with a length of 658 bp (GenBank 1 College of Life Science, Nanchang University, accession number: KF492154). There is only one study Nanchang 330031, China about mitochondrial phylogenomics and genetic relation- 2 Plant Protection Unit, Dayu County, Ganzhou 341500, China ships of Dendrolimus taxa (Dendrolimus punctatus, 123 Author's personal copy 676 Genetica (2016) 144:675–688 Dendrolimus spectabilis and Dendrolimus tabulaeformis) family. Also, it is a general knowledge of the genetic using whole mitochondrial genomes (Qin et al. 2015), but relationship between Lasiocampidae and other superfami- no other molecular studies about Lasiocampidae species. lies in Lepidoptera. Although T.vishnou guttata taxonomic classification is certain, the classification of Lasiocampidae is not uniform. Some previous molecular studies by Minet (1990) and Materials and methods Zwick et al. (2011) supports that Lasiocampidae should be a independent group separating from Bombycoidea. The Sample collection and DNA extraction genetic information of T.vishnou guttata is important to know the genetic diversity of Lasiocampidae, also the Adult of T. vishnou guttata had been collected from Tao- phylogenetic relationships within the families Bomby- hong Ridge National Sika Deer Nature Reserve coidea basing on mitogenome. Therefore, it is significant to (116°4201000E, 29°470700N), Jiujiang City, Jiangxi Province, acquire information of complete mitogenome of T. vishnou China. The morphological feature of living adult had been guttata. Phylogenetic analyses aims to contribute to know observed with inverted microscope (Leica, Germany). The genetic relationship and different of morphological classi- species identification had been on the basis of authoritative fication within lepidopteran species. Similar with previous literature: Fauna Sinica (Liu and Wu 2006). The samples studies, 13 PCGs nucleotide sequences are performed to had been preserved in 95 % ethanol, and stored at -40 °C establish phylogenetic analyses resulting from their refrigerator until DNA extraction. Total genomic DNA had stable conservative nature (Hong et al. 2008; Feng et al. been extracted from single specimen using GENERAY 2010), which show each single origin of superfamilies and BIOTECH Genomic DNA kit (Shanghai, China). support the traditional morphological classification methods. PCR amplification and sequencing Mitochondrial DNA is involved in the protein synthesis, transcription, and replication, which have higher research To amplify the entire mitogenome of T. vishnou guttata,10 values (Chen et al. 2012). Mitochondrial genomes (mi- pair of primers (Table 1S) had been designed using Primer togenomes) with characteristics of no recombination, Premier 5 (Lalitha 2000). Five universal primers were used maternal inheritance and higher mutation rate are beneficial for PCR-amplified short ragments of the mitogenome of T. to check out the gene changes in a relatively short period and vishnou guttata (Lee et al. 2006; Simon et al. 2006). compare the differences of the same gene in different spe- According to the five short fragments, we had design the cies. It is widely applied in phylogenetic analyses, popula- primers of long fragments and amplified all the fragments. tion genetics, spectrum geography, evolutionary genomics, PCR amplification had been carried out in 25 lL reactions evolutionary biology and genetic population studies (Avise that contained 17 lL ddH2O, 2.5 lL 10X LA PCR Buffer et al. 1987; Bae et al. 2004). The mitogenomes of lepi- II (Mg2? plus), 2.5 lL dNTP Mixture (2.5 mM), 0.25 lL dopteran insects were a typical molecule of circular stucture LA Taq (5 U/lL, TaKaRa, Dalian, China), 1.0 lL DNA with a length of 14–19 kb, including 13 PCGs, 2 rRNA template (200 ng/lL) and 1.0 lL of each primer (10 mM). genes, 22 tRNA genes, and an A ? T-rich region (Gray et al. PCR conditions were 94 °C for 3 min; 35 cycles of 30 s at 1999), which contains genome replicative and transcrip- 94 °C, 30 s at 50–60 °C, 1–3 min at 72 °C; and 72 °C for tional initiation sites (Boore 1998; Taanman 1999). Lepi- 10 min. The PCR products had been detected by 1.0 % doptera insects have a profound effect on human society, agarose-gel electrophoresis. Then, we had obtain all the whether functioning as major pests of agriculture and for- sequences of complete mitogenome after all PCR products estry, or as pollinators and economic insects (Wang et al. were sequenced by Sangon Biotech (Shanghai, China). 2013). Therefore, a thorough understanding of the mito- genomes applied to studying the origin, evolution and phy- Sequences analyses and gene annotation logenetic relationship is quite important to control lepidopteran pests or use economic insects (Goldsmith et al. According to the sequence similarity of other Lasiocamp- 2005; Kristensen et al. 2007). idae insects, all the sequences had been assembled and In this study, the complete mitogenome sequence of T. proofread using the program DNAStar software and the vishnou guttata is sequenced and verified firstly (GenBank nucleotide blast (http://www.ncbi.nlm.nih.gov/blast) on the accession number: KU884483) accoding to the character- NCBI database. 13 PCGs had been identified in compara- istics of closely related species mitogenomes published in tion with the complete mitogenomes of other Lasiocamp- NCBI. The position of T. vishnou guttata in the phylogeny idae insects published on NCBI by Clustal X 1.83 software reflects genetic relationship with the species of known (Thompson et al. 1997) and the ORF finder (http://www. mitogenomes, especially the species in Lasiocampidae ncbi.nlm.nih.gov/orf/gorf.html).
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