Abstract

Simple SummaryTyphlocybinae is the smallest sized and most evolved leafhopper, body length 2–4 mm, forewing with four apical cells but lacking closed preapical cells. It comprises over 6000 species (Dietrich, 2013) distributed worldwide, making it the second largest group of Cicadellidae. Previous phylogenetic analyses in this subfamily were mainly based on morphological characters and were restricted to several gene fragments. To provide further insight into the relationships of its included tribes, complete mitogenomes of two Alebrini species (Shaddai acuminatus, Sobrala sp.), two Dikraneurini species (Dikraneura (Dikraneura) zlata, Robusta emeiensis), two Empoascini species (Alebroides salici, Empoasca serrata), two Erythroneurini species (Elbelus tripunctatus, Kaukania anser), two Typhlocybini species (Eupteryx (Eupteryx) adspersa, Eurhadina jarrary), and one Zyginellini species (Yangisunda tiani) are newly sequenced and comparatively analyzed. The mitogenomes comprise the typical set of 37 mitochondrial genes and a large non-coding region (A+T-rich region). The acceptor arm of trnR is the most inconstant among all the tRNAs, due to the acceptor arm comprising unpaired bases. Phylogenetic analyses using Bayesian inference and maximum likelihood methods produced a well-resolved framework of Typhlocybinae, showed the monophyly of Typhlocybinae and its inner tribes, except for Typhlocybini and Zyginellini combined. These results provide the valuable data toward the future study of the phylogenetic relationships in this subfamily.To explore the characteristics of mitogenomes and discuss the phylogenetic relationships and molecular evolution of the six tribes within Typhlocybinae, 11 complete mitogenomes are newly sequenced and comparatively analyzed. In all of these complete mitogenomes, the number and order of the genes are highly conserved in overall organization. The PCGs initiate with ATN/TTG/GTG and terminate with TAA/TAG/T. Almost all tRNAs are folded into the typical clover-leaf secondary structure. The control region is always variable in length and in numbers of multiple tandem repeat units. The atp8 and nad2 exhibits the highest evolution rate among all the PCGs. Phylogenetic analyses based on whole mitogenome sequences, with three different datasets, using both maximum likelihood and Bayesian methods, indicate the monophyly of Typhlocybinae and its inner tribes, respectively, except for Typhlocybini and Zyginellini that are paraphyletic. Finally, we confirm that Erythroneurini is a subtribe of Dikraneurini.

Highlights

  • IntroductionThe mitogenome provides genome-level information, including base composition, sequence arrangement, codon usage or variation, RNA secondary structures and control region characters

  • Licensee MDPI, Basel, Switzerland.Members of Typhlocybinae are in the family Cicadellidae, Auchenorrhyncha, and order Hemiptera

  • Typhlocybinae is composed of six tribes (Alebrini, Dikraneurini, Empoascini, Erythroneurini, Typhlocybini and Zyginellini) in the most well-accepted taxonomy system that was proposed by Dworakowska in 1979 [3]

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Summary

Introduction

The mitogenome provides genome-level information, including base composition, sequence arrangement, codon usage or variation, RNA secondary structures and control region characters. We use three different datasets (PCG123, PCG123R, PCG12) and use both maximum likelihood (ML) and Bayesian inference (BI) methods to reconstruct their phylogenetic relationships and provide further insight into their taxonomic status This is based on 11 newly sequenced and functional annotated complete mitogenomes of two Alebrini species (Shaddai sp., Sobrala sp.), two Dikraneurini species, (Dikraneura (Dikraneura) zlata, Dikraneurini sp.), two Empoascini species (Alebroides salicis, Empoasca serrata), two Erythroneurini species (Elbelus tripunctatus, Kaukania anser), two Typhlocybini species (Eupteryx (Eupteryx) adspersa, Eurhadina jarrary), and one Zyginellini species (Yangisunda tiani) along with 13 previously available mitogenomes in GenBank. The genomic size, base composition and skewness, sequence arrangements, codon usage or variation, evolutionary rate, genetic distance, ka/ks, start and stop codons, RNA secondary structures and control region characters are analyzed

Sample Preparation and Genomic DNA Extraction
Sequence Alignment and Phylogenetic Analysis
Mitogenome Structure and Nucleotide Composition
Protein-Coding Genes and Codon Usage
Transfer and Ribosomal
Control Region
Gene Overlaps and Intergenic Spacers
Predicted
Nucleotide Diversity and Evolutionary Rate Analysis
Phylogenetic Relationships
Findings
Conclusions
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