Research Article
Research Article
First mitochondrial genome of subfamily Julodinae (Coleoptera, Buprestidae) with its phylogenetic implications
expand article infoZhonghua Wei, Xuyan Huang, Aimin Shi
‡ China West Normal University, Nanchong, China
Open Access


Complete mitochondrial genomes of three species of the family Buprestidae were sequenced, annotated, and analyzed in this study. To explore the mitogenome features of the subfamily Julodinae and verify its phylogenetic position, the complete mitogenome of Julodis variolaris was sequenced and annotated. The complete mitogenomes of Ptosima chinensis and Chalcophora japonica were also provided for the phylogenetic analyses within Buprestidae. Compared to the known mitogenomes of Buprestidae species varied from 15,499 bp to 16,771 bp in length, three newly sequenced mitogenomes were medium length (15,759–16,227 bp). These mitogenomes were encoded 37 typical mitochondrial genes. Among the three studied mitogenomes, Leu2 (L2), Ser2 (S2), and Pro (P) were the three most frequently encoded amino acids. Within the Buprestidae, the heterogeneity in sequence divergences of Agrilinae was highest, whereas the sequence homogeneity of Chrysochroinae was highest. Moreover, phylogenetic analyses were performed based on nucleotide matrix (13 PCGs + 2 rRNAs) among the available sequenced species of Buprestidae using Bayesian Inference and Maximum Likelihood methods. The results showed that the Julodinae was closely related to the subfamily Polycestinae. Meanwhile, the genera Melanophila, Dicerca, and Coomaniella were included in Buprestinae, which was inconsistent with the current classification system of Buprestidae. These results could contribute to further studies on genetic diversity and phylogeny of Buprestidae.


Jewel beetles, Julodinae, mitogenome, phylogenetics


The family Buprestidae is one of the largest families in Coleoptera, including six subfamilies, 521 genera, and more than 15,000 species distributed worldwide (Bellamy 2008; Kubáň et al. 2016). In this family, all species are phytophagous. The adults are feeders on flowers, leaves and stems, whereas the larvae are internal feeders in roots and stems, or feed on the foliage of woody and herbaceous plants, the larvae of Julodinae are soil habitants feeding externally by the roots (Bellamy and Volkovitsh 2016). Different groups have different functions covered ecological, social and economic functions, such as: most larvae of Buprestinae and Chrysochroinae are important decomposers of woody plants; with most species being ornamental beetles with attractive metallic luster; many species of Agrilinae are forest and agricultural pests; and some species of the tribes Stigmoderini, Acmaeoderini, and Anthaxiini are pollinator taxa. Although some buprestid taxonomists have made important contributions to the classification based on morphological analyses (Cobos 1980, 1986; Tôyama 1987; Hołyński 1988, 1993, 2009; Kolibáè 2000; Bellamy 2003), the problems of the overall classification of Buprestidae remain.

In the past two decades, the mitochondrial genome emerged as important molecular data for higher-level phylogenetic analyses (Saccone et al. 1999; Timmermans et al. 2010, 2016; Cameron 2014; Li et al. 2015; Qin et al. 2015; Nie et al. 2020, 2021; Motyka et al. 2022; Zheng et al. 2022), evolutionary strategies (Krzywinski et al. 2011; Nie et al. 2019; Motyka et al. 2022; Zhang et al. 2022), and genetic diversity analyses (Lim et al. 2021). The buprestid mitogenome also caught the attention of taxonomists. In Buprestidae, the first complete mitogenome of Chrysochroa fulgidissima (Schönherr, 1817) was reported by Hong et al. (2009). In the same year, the mitogenome of Acmaeodera sp. was used to analyze the nonstationary evolution and compositional heterogeneity of Coleoptera. To date, only 22 buprestid mitogenomes (Table 1) have been reported worldwide, including three newly generated in this study.

Table 1.

Information on the mitogenomes of Buprestidae and outgroup taxa used for phylogenetic analysis.

Subfamily Taxa Accession No. Genome size (bp) A+T% AT-skew Reference
Agrilinae Coraebus diminutus Gebhardt, 1928 OK189521 15,499 68.42 0.12 Wei 2022
Coraebus cloueti Théry, 1895 OK189520 15,514 69.27 0.11 Wei 2022
Coraebus cavifrons Descarpentries & Villiers, 1967 MK913589 15,686 69.79 0.12 Cao and Wang 2019a
Meliboeus sinae Obenberger, 1935 OK189522 16,108 72.42 0.11 Wei 2022
Sambus femoralis Kerremans, 1892 OK349489 15,367 73.23 0.12 Wei 2022
Agrilus sichuanus Jendek, 2011 OK189519 16,521 71.73 0.12 Wei 2022
Agrilus planipennis Fairmaire, 1888 KT363854 15,942 71.90 0.12 Duan et al. 2017
Agrilus mali Matsumura, 1924 MN894890 16,204 74.46 0.08 Sun et al. 2020
Trachys auricollis Saunders, 1873 MH638286 16,429 71.05 0.10 Xiao et al. 2019
Trachys troglodytiformis Obenberger, 1918 KX087357 16,316 74.62 0.10 Unpublished
Trachys variolaris Saunders, 1873 MN178497 16,771 72.11 0.11 Cao and Wang 2019b
Buprestinae Melanophila acuminata (De Geer, 1774) MW287594 15,853 75.66 0.02 Peng et al. 2021
Anthaxia chinensis Kerremans, 1898 MW929326 15,881 73.61 0.09 Chen et al. 2021
Coomaniella copipes Jendek & Pham, 2013 OL694145 16,196 74.47 0.03 Huang et al. 2022
Coomaniella dentata Song, 2021 OL694144 16,179 76.59 0.01 Huang et al. 2022
Chrysochroinae Chrysochroa fulgidissima (Schönherr, 1817) EU826485 15,592 69.92 0.15 Hong et al. 2009
Chalcophora japonica (Gory, 1840) OP388437 15,759 67.97 0.13 In this study
Chalcophora japonica (Gory, 1840) OM161962 15,759 67.94 0.13 Weng et al. 2022
Dicerca corrugata Fairmaire, 1902 OL753086 16,276 71.76 0.09 Huang et al. 2022
Polycestinae Acmaeodera sp. FJ613420 16,217 68.41 0.11 Sheffield et al. 2009
Ptosima chinensis Marseul, 1867 OP388449 16,115 67.00 0.13 In this study
Julodinae Julodis variolaris (Pallas, 1771) OP390084 16,227 70.43 0.12 In this study
outgroup Heterocerus parallelus Gebler, 1830 KX087297 15,845 74.03 0.13 Unpublished
Dryops ernesti Gozis, 1886 KX035147 15,672 72.98 0.07 Unpublished

To date, the mitogenome of the subfamily Julodinae has not been reported. The lack of the data on complete mitogenome of Julodinae species has limited our understanding of the real phylogenetic relationships within jewel beetles. The single molecular phylogenetic analysis, including Julodinae, showed that Julodinae is monophyletic group and close to Polycestinae (Evans et al. 2015). The subfamily Julodinae includes one tribe and six genera (Hołyński 2014). The described Julodinae species are mainly distributed in the arid and semiarid zones of the Ethiopian and Palaearctic regions, except for the species of the genus Sternocera Eschscholtz, 1829 distributed in humid tropical zones of Asia and Africa (Bellamy 2008; Hołyński 2014).

In the present study, three complete mitogenomes are sequenced and annotated, of which that of Julodis variolaris (Pallas, 1771) is the first complete mitogenome sequence to be reported in the subfamily Julodinae. In China, this species is widely distributed in Xinjiang Uygur Autonomous Region. The adults, appearing in May and June, feeder on the leaves of Haloxylon ammodendron (Meyer, 1829) and the larvae feeder on the roots of this plant. Additionally, the complete mitogenomes of Chalcophora japonica (Gory, 1840) (Chrysochroinae: Chalcophorini) and Ptosima chinensis Marseul, 1867 (Polycestinae: Ptosimini) are provided for phylogenetic analyses, which are also enriching the diversity of mitogenomes studied in Buprestidae. The total length of the mitogenome in C. japonica was consistent with the results of Weng et al. (2022). In order to explore the phylogenetic position of the subfamily Julodinae, phylogenetic analyses of the family Buprestidae were performed based on a nucleotide matrix (13 PCGs + 2 rRNAs) among buprestid species using Bayesian Inference (BI) and Maximum Likelihood (ML) methods.

Materials and methods

Sampling and DNA extraction

Specimens of J. variolaris were collected on H. ammodendron in the vicinities of Turpan City, Xinjiang Uygur Autonomous Region, China, on 14 May 2022. Specimens of P. chinensis were collected from Dayaoshan Mountains in Guangxi Zhuang Autonomous Region, China, on 20 March 2021. Specimens of C. japonica were collected from Quanzhou City, Fujian Province, China, on 23 February 2021. The above specimens are preserved in 95% alcohol at -24 °C in specimen collection at China West Normal University, Nanchong, China. Next-generation sequencing and assembly were performed by Beijing Aoweisen Gene Technology Co. Ltd. (Beijing, China) to obtain the complete mitogenome sequences.

Sequence assembly, annotation, and analysis

The raw data were processed using Trimmomatic v. 0.35 (Bolger et al. 2014) to remove low-quality reads and obtain a high-quality clean data. Finally, 4.8 Gb, 5.28 Gb, and 6.8 Gb clean data were obtained to assemble complete mitogenome of J. variolaris, P. chinensis, and C. japonica, respectively. Three mitogenome sequences were annotated using Geneious 11.0.2 (Kearse et al. 2012) based on the invertebrate mitochondrial genetic code. All tRNA genes were reconfirmed using the online tool MITOS Web Server (Bernt et al. 2013) and the second structures were further predicted using tRNAscan-SE server v. 1.21 (Lowe and Chan 2016). Two rRNA genes were identified by alignment with other buprestid rRNA sequences. Three mitogenome maps were drawn using Organellar Genome Draw v. 1.3.1 (Greiner et al. 2019). Strand asymmetry of mitogenome sequence was calculated using the formulae reported by Perna and Kocher (1995): AT-skew = (A – T)/(A + T), and GC-skew = (G – C)/(G + C). The base composition and relative synonymous codon usage (RSCU) values of three mitogenome sequences were determined using MEGA v. 12.0.0 (Kumar et al. 2016). The non-synonymous substitutions (Ka) and synonymous substitutions (Ks) of all PCG genes were calculated using DnaSP v. 5 (Librado and Rozas 2009). The tandem repeat elements of control region (CR, also known as A + T-rich region) were detected by the online tool Tandem Repeats Finder (Benson 1999). The heterogeneous analysis of nucleotide matrix (13 PCGs + 2 tRNAs) was performed using AliGROOVE v. 1.06 (Kück et al. 2014).

Phylogenetic analysis

To investigate mitogenome arrangement patterns in Buprestidae, the gene orders of all known buprestid mitogenomes were compared with that of closely related taxa. A total of 22 buprestid mitogenomes (Table 1), including three newly generated sequences in this study, were subjected for phylogenetic analyses, using Heterocerus parallelus Gebler, 1830 (Heteroceridae) and Dryops ernesti Gozis, 1886 (Dryopidae) as outgroups (Xiao et al. 2019; Huang et al. 2022; Wei 2022). The test of substitution saturation for the dataset (13 PCGs + 2 rRNAs) was performed with DAMBE to test whether the sequence is suitable for constructing a phylogenetic tree (Xia 2017). Then, the phylogenetic trees were reconstructed using nucleotide matrix 13 PCGs + 2 rRNAs based on ML and BI methods. The nucleotide matrix was aligned using ClustalW (Thompson et al. 1994) and trimmed by trimAl v. 1.2 (Capella-Gutiérrez et al. 2009). In BI and ML analyses, the best-fit models were deduced by ModelFinder (Kalyaanamoorthy et al. 2017). The phylogenetic trees were reconstructed using IQ-tree v. 1.6.8 (Guindon et al. 2010) and MrBayes v. 3.2.6 (Ronquist et al. 2012) integrated into PhyloSuite v. 1.2.2 (Zhang et al. 2020). During this analyzing process, PhyloSuite was run with previous parameters (Wei 2022).


Genome organization and base composition

We sequenced and annotated the complete mitogenome of J. variolaris (GenBank No. OP390084), P. chinensis (No. OP388449), and C. japonica (No. OP388437). Overall, these mitogenome sequences were 15,759 to 16,227 bp in length, which are medium length in Buprestidae (Table 1). It is a circular, double-stranded ring that includes 37 insect mitochondrial genes (13 PCGs, 22 tRNAs, and 2 rRNAs) and an A + T-rich region (control region, CR).

In these three mitogenome, the N-strand encoded the sense-strand of 14 genes (nad1, nad4L, nad4, nad5, trnQ, trnV, trnL1, trnP, trnH, trnF, trnY, trnC, rrnL, and rrnS), while the J-strand encoded the sense-strand of the remaining 23 genes (Table 2), which was consistent with the known buprestid species (Cao and Wang 2019a, b; Xiao et al. 2019; Chen et al. 2021; Peng et al. 2021; Huang et al. 2022; Wei 2022; Weng et al. 2022).

Table 2.

The three newly annotated Buprestidae mitogenomes. The order of the three species in the table is as follows: Julodis variolaris, Ptosima chinensis, and Chalcophora japonica. – not determined.

Gene Strand Position From To Start codons Stop codons Intergenic nucleotides
trnI J 1/1/1 66/64/67 0/0/0
trnQ N 64/65/65 134/133/133 -3/0/-3
trnM J 134/133/133 202/201/201 -1/-1/-1
nad2 J 203/202/202 1228/1221/1224 ATT/ATT/ATC TAA/TAA/TAA 0/0/0
trnW J 1241/1220/1223 1306/1285/1291 12/-2/-2
trnC N 1299/1278/1284 1360/1341/1345 -7/-7/-7
trnY N 1361/1343/1346 1426/1406/1409 0/1/0
cox1 J 1428/1408/1411 2958/2941/2943 –/–/– T(AA)/T(AA)/TAA 1/1/1
trnL2 J 2959/2942/2944 3024/3006/3009 0/0/0
cox2 J 3025/3007/3010 3709/3691/3697 ATA/ATA/ATA T(AA)/T(AA)/T(AA) 0/0/0
trnK J 3710/3692/3698 3780/3761/3767 0/0/0
trnD J 3780/3762/3768 3845/3824/3829 -1/0/0
atp8 J 3846/3825/3887 4004/3983/4042 ATT/ATT/ATT TAA/TAA/TAA 0/0/57
atp6 J 3998/3977/4036 4672/4651/4710 ATG/ATG/ATG TAA/TAA/TAA -6/-7/-7
cox3 J 4672/4651/4710 5458/5439/5496 ATG/ATG/ATG T(AA)/TAA/T(AA) -1/-1/-1
trnG J 5459/5447/5497 5522/5512/5558 0/7/0
nad3 J 5523/5513/5559 5876/5866/5912 ATT/ATT/ATT TAG/TAG/TAG 0/0/0
trnA J 5875/5865/5911 5940/5929/5974 -2/-2/-2
trnR J 5940/5934/5975 6006/5998/6035/ -1/4/0
trnN J 6006/6002/6035 6070/6066/6099 -1/3/-1
trnS1 J 6071/6067/6100 6137/6131/6166 0/0/0
trnE J 6138/6132/6168 6201/6197/6229 0/0/1
trnF N 6201/6196/6229 6265/6260/6292 -1/-2/-1
nad5 N 6265/6260/6293 7983/7978/8012 ATA/ATC/GTG TAA/TAA/T(AA) -1/-1/0
trnH N 7984/7979/8013 8047/8042/8075 0/0/0
nad4 N 8048/8042/8076 9380/9379/9411 ATG/ATG/ATG T(AA)/TAA/T(AA) 0/-1/0
nad4L N 9374/9373/9405 9664/9666/9695 ATG/ATG/GTG TAA/TAA/TAA -7/-7/-7
trnT J 9667/9669/9698 9731/9733/9762 2/2/2
trnP N 9731/9733/9763 9795/9798/9827 -1/-1/0
nad6 J 9797/9800/9829 10,303/10,306/10,335 ATA/ATA/ATC TAA/TAA/TAA 1/1/1
cytb J 10,303/10,306/10,335 11,454/11,448/11,474 ATG/ATG/ATG TAG/TAA/TAG -1/-1/-1
trnS2 J 11,453/11,447/11,473 11,519/11,512/11,539 -2/-2/-2
nad1 N 11,539/11,536/11559 12,489/12,480/12,509 TTG/TTG/TTG TAA/TAA/TAG 39/33/19
trnL1 N 12,491/12,482/12,511 12,554/12,546/12,574 1/1/1
rrnL N 12,555/12,547/12,575 13,855/13,845/13,873 0/0/0
trnV N 13,856/13,846/13,847 13,925/13,915/13,943 0/0/-27
rrnS N 13,926/13,916/13,944 147,17/14,664/14,679 0/0/0
A+T-rich region 14,718/14,665/14,680 16,227/16,115/15,759 0/0/0

These three mitogenome sequences had a high A + T content, with an average of 68.47%, showing a strong AT bias (Suppl. material 1: table S1). Among them, the A + T content of J. variolaris (70.43%) was higher than of both C. japonica (67.97%) and P. chinensis (67.00%). These three mitogenome sequences showed a positive AT skew (0.12–0.13) and negative GC skew (-0.22), which is consistent with the known buprestid species. In this study, there were 21 gaps in three mitogenome sequences, which varied from 1 bp to 57 bp. The longest intergenic spacer (bp) was located between trnD and atp8 genes in C. japonica. There were 41 overlapping gene regions in total, ranging from 1 bp to 27 bp in length.

Protein-coding genes, codon usage, and nucleotide diversity

In Julodinae, the concatenated length of 13 PCGs of J. variolaris (Julodinae) was 11,170 bp, which encoded 3715 amino acid residues. In P. chinensis (Polycestinae), the total length of 13 PCGs was 11,162 bp, which encoded 3710 amino acid residues. In C. japonica (Chrysochroinae), the total length of 13 PCGs was 11,161 bp, which encoded 3710 amino acid residues. Compared with the other known buprestid species (Chen et al. 2021; Peng et al. 2021; Huang et al. 2022; Wei 2022; Weng et al. 2022), the concatenated length of 13 PCGs and the number of amino acid-coding codons of Julodinae is slightly higher than in other subfamilies.

The majority of PCGs directly used ATN as the start codon, but the exceptions were nad1 (J. variolaris, P. chinensis, and C. japonica), nad4L (C. japonica), and nad5 (C. japonica) genes which started with TTG, GTG, and GTG, respectively. The unusual start codon TTG was also reported in Agrilinae (Wei 2022) and Buprestinae (Huang et al. 2022). The start codon of the cox1 gene in these three mitogenomes was not determined, which may use non-canonical start codons (Friedrich and Muquim 2003; Fenn et al. 2007; Yang et al. 2013; Wang et al. 2021; Wu et al. 2022). There were three types of stop codons, TAA, TAG, and an incomplete stop codon T, which was completed by the addition of 3’ A residues to the mRNA.

To investigate further, the frequency of synonymous codon usage and relative synonymous codon usage (RSCU) values were calculated and presented. Taken together, the three most frequently used amino acids were L2, S2, and P (Fig. 1A, B), and the most frequently used codons were TTA (L2), TCT (S2), and CCT (P) (Fig. 2).

Figure 1. 

Numbers of different amino acids in the three new mitogenome sequences A and the percentages of the top ten amino acids B the stop codon is not included in these graphs.

Figure 2. 

Relative synonymous codon usage (RSCU) of the three newly sequenced mitogenomes.

The Ka/Ks ratio can be used to estimate whether a sequence is undergoing negative, neutral, or positive selection (Hurst 2002; Mori and Matsunami 2018). The ratio of Ka/Ks for each mitogenome sequence was calculated using Anthaxia chinensis Kerremans, 1898 as the reference sequence (Fig. 3A). In three mitogenome sequences, values of Ka, Ks, and Ka/Ks were all less than 1, suggesting the presence of purifying selection in these three species.

Figure 3. 

Evolutionary rates of mitochondrial genomes in three new mitogenome sequence (A) and the heterogeneity of two dataset in Buprestidae (B).

Ribosomal and transfer RNA genes, and heterogeneity

The rRNA genes were located between the A + T-rich region and trnL1, and separated by trnV, which is consistent with previous studies (Duan et al. 2017; Cao and Wang 2019a, b; Xiao et al. 2019; Sun et al. 2020; Chen et al. 2021; Peng et al. 2021; Huang et al. 2022; Wei 2022; Weng et al. 2022). The total length of rRNA genes ranged from 2035 bp (C. japonica) to 2093 bp (J. variolaris), of which the length of 16S gene ranged from 1299 bp (C. japonica and P. chinensis) to 1301 bp (J. variolaris). The A + T content of rRNA genes ranged from 71.50% (C. japonica) to 74.30% (J. variolaris).

The concatenated lengths of all tRNA genes ranged from 1437 bp (C. japonica) to 1456 bp (J. variolaris), whereas individual tRNA genes ranged from 61 bp (trnR) to 71 bp (trnK), of which eight tRNA genes were encoded on the N-strand and the remaining 14 genes encoded on the J-strand. The predicted secondary structure of tRNAs showed a standard clover-leaf structure (Suppl. material 1: figs S2–S4), except for trnS1 (Fig. 4A), which lacked the dihydrouridine arm, and formed a loop commonly found in other insects (Xiao et al. 2011; Park et al. 2012; Yu et al. 2016; Yan et al. 2017; Yu and Liang 2018; Li et al. 2019). The UG mismatches were detected in some tRNAs (Suppl. material 1: figs S2–S4), which also appeared in other buprestid species (Sun et al. 2020; Chen et al. 2021; Huang et al. 2022; Wei 2022; Weng et al. 2022).

Figure 4. 

The predicted secondary cloverleaf structure for the trnS1 of three new mitogenomes (A) and the gene order of known buprestid mitogenomes (B).

The degree of heterogeneity of the PCGs + RNAs dataset was higher than that of the PCGs dataset (Fig. 3B). Additionally, the heterogeneity in sequence divergences was slightly stronger for Agrilinae than for other families (Fig. 3B). The heterogeneity in sequence homogeneity was higher for Chrysochroinae than other families.

A + T-rich region and gene arrangement

The A + T-rich region was the largest non-coding region in mitogenome, located between trnI and rrnS. This region, containing regulatory elements correlated with the regulation of replication and transcription (Zhang et al. 1995), plays a very important role in molecular evolution (Zhang and Hewitt 1997). The length of A + T-rich region ranged from 1080 bp (C. japonica) to 1510 bp (J. variolaris), which are of medium length in the Buprestidae (Sun et al. 2020; Huang et al. 2022; Wei 2022). The A + T content of the A + T-rich region of C. japonica (75.93%) and P. chinensis (78.38%) was found to be higher than that of the whole genome (67.97%, 67.00%), PCGs (66.46%, 64.55%), rRNAs (71.50%, 72.51%), and tRNAs (68.82%, 71.46%), whereas the A + T content of J. variolaris (72.85%) was lower than that of whole genome (70.43%), PCGs (68.82%), rRNAs (74.30%), and tRNAs (74.79%).

The tandem repeat regions of three species were detected in this study. The repeat regions in each of the three new mitogenomes differ from each other in length and copy number of tandem repeat units. The repeat region of J. variolaris was 43 bp in length, comprising a 17 bp and a 26 bp tandem repeat element. In contrast, in P. chinensis, the total length of the repeat sequence was 111 bp, consisting of three incomplete repeat units. These tandem repeat elements are slightly shorter than those of Agrilinae (Wei 2022).

The gene rearrangements were regarded as important molecular markers for exploring the evolution and phylogeny of insects (Dowton et al. 2002; Cameron 2014). All the buprestid mitogenomes released in GenBank were compared and analyzed, with one mitogenome arrangement pattern exhibited in Buprestidae (Fig. 4B). The mitochondrial gene order of these three species was consistent with other known buprestid mitogenomes.

Phylogenetic analysis

For the concatenated sequences, the test of substitution saturation showed that the value of Iss = 0.3910 was significantly smaller than Iss.c = 0.8537 and p (0.0000) < 0.01, suggesting the sequences suitable for phylogenetic analysis. In the present study, both ML and BI trees using a nucleotide matrix (13 PCGs + 2 rRNAs) produced identical topologies (Fig. 5, Suppl. material 1: fig. S5), (Chrysochroniae + ((Julodinae + Polycestinae) + Buprestinae) + Agrilinae), in terms of subfamily-level relationship.

Figure 5. 

Phylogenetic relationships of studied species of Buprestidae using BI analyses based on 13 PCGs + 2 rRNAs of mitogenomes. The numbers on the branches show posterior probabilities.

The target species J. variolaris, representing Julodinae, formed an independent clade close to Polycestinae with high support values (BI: 1; ML: 94), which supported the results of a previous study (Evans et al. 2015). The target species P. chinensis and Acmaeodera sp. are grouped together as an independent clade with high support values (BI: 1; ML: 100), representing Polycestinae. The Julodinae and Polycestinae formed a clade which was sister to Buprestinae with high support values (BI: 1; ML: 84). The target species C. japonica was clustered with other chrysochroine species as a clade, representing Chrysochroinae, with high support values (BI: 1; ML: 100). All the species of Agrilinae were clustered on one branch with high support values (BI: 1; ML: 100) and close to other buprestid clades, while the Coraebini was polyphyletic.


The gene composition and arrangement of these three mitogenomes are the same as other known buprestid mitogenomes (Cao and Wang 2019a, b; Xiao et al. 2019; Chen et al. 2021; Peng et al. 2021; Huang et al. 2022; Wei 2022; Weng et al. 2022). These three mitogenome had a positive AT skew, which was similar to most known buprestid mitogenomes (Duan et al. 2017; Cao and Wang 2019a, b; Xiao et al. 2019; Sun et al. 2020; Chen et al. 2021; Peng et al. 2021; Huang et al. 2022; Wei 2022; Weng et al. 2022). The genes nad1 (J. variolaris, P. chinensis, and C. japonica), nad4L, and nad5 (C. japonica) which started with TTG, GTG, and GTG, respectively, was also reported by previous studies in Buprestidae (Huang et al. 2022; Wei 2022). The Julodinae are closest to Polycestinae with high support values, which is consistent with the results of a previous study (Evans et al. 2015). The monophyly of Buprestidae has been corroborated once more, as all the buprestid species converge together as an independent clade (Evans et al. 2015; Huang et al. 2022; Wei 2022). In this study, the Coraebini was also found to be polyphyletic with the genera Meliboeus Deyrolle, 1864 and Coraebus Gory & Laporte, 1839 in different clades, also consistent with the previous studies (Evans et al. 2015; Huang et al. 2022; Wei 2022). Compared to Melanophilini, Coomaniellini is more closely related to Dicercini, which is in line with previous studies (Volkovitsh 2001; Evans et al. 2015; Huang et al. 2022).

In the present study, the sampling might be too limited to address the comprehensive phylogeny of Buprestidae. In the future, classification problems could be solved when enough mitogenomes are accumulated for more buprestid species, which requires the cooperation of taxonomists around the world.


In this study, the complete mitogenomes of Julodis variolaris, Chalcophora japonica, and Ptosima chinensis were annotated and analyzed, of which the mitogenome of J. variolaris was the first complete mitogenome representative of the subfamily Julodinae. The three mitogenome sequences were of medium length (15,759–16,227 bp) in Buprestidae. These three mitogenomes shared the same gene order, which was consistent with those of known buprestid species. These three mitogenome sequences all had a high A + T content, and strong AT bias. All PCGs of the three species began with the typical ATN codon except nad1 (J. variolaris, P. chinensis, and C. japonica), nad4L (C. japonica), and nad5 (C. japonica) which were initiated with TTG, GTG, and GTG, respectively. In the present study, the BI and ML trees had exact same topologies with high-value support. The results of phylogenetic analyses also show that Julodinae is close to Polycestinae, the clade composed of Julodinae and Polycestinae is close to that of Buprestinae, and the Agrilinae clade is sister to that of (Chrysochroniae + ((Julodinae + Polycestinae) + Buprestinae)), and all the subfamilies are grouped in a monophyletic group with high support.


We thank Dr. Mark Volkovitsh (Russian Academy of Sciences, Moscow, Russia) and Dr. Zhao Pan (Hebei University, Baoding, China) for revising the manuscript. This work was supported by Natural Science Foundation of Sichuan Province (2022NSFSC1707) and the Doctoral Scientific Research Foundation of China West Normal University (20E054).


  • Bellamy CL (2003) An illustrated summary of the higher classification of the superfamily Buprestoidea (Coleoptera). Folia Heyrovskyana, Supplementum 10: 1–197.
  • Bellamy CL (2008) A world catalogue and bibliography of the jewel beetles (Coleoptera: Buprestoidea), Volumes 1–4. Pensoft Series Faunistica No. 76–79, Sofia/Moscow, 2684 pp.
  • Bellamy CL, Volkovitsh M (2016) 18 Buprestoidea Crowson, 1955. In: Beutel RG, Leschen RAB (Eds) Handbook of Zoology, Arthropoda: Insecta, Morphology and Systematics (Archostemata, Adephaga, Myxophaga, Polyphaga partim) (2nd edn. ). Walter de Gruyter, Berlin/Boston, 543–552.
  • Bernt M, Donath A, Jühling F, Externbrink F, Florentz C, Fritzsch G, Pütz J, Middendorf M, Stadler PF (2013) MITOS: Improved de novo metazoan mitochondrial genome annotation. Molecular Phylogenetics and Evolution 69(2): 313–319.
  • Cao LM, Wang XY (2019a) The complete mitochondrial genome of the jewel beetle Coraebus cavifrons (Coleoptera: Buprestidae). Mitochondrial DNA Part B Resources 4(2): 2407–2408.
  • Cao LM, Wang XY (2019b) The complete mitochondrial genome of the jewel beetle Trachys variolaris (Coleoptera: Buprestidae). Mitochondrial DNA Part B Resources 4(2): 3042–3043.
  • Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T (2009) TrimAl: A tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 25(15): 1972–1973.
  • Chen B, Wei ZH, Shi AM (2021) The complete mitochondrial genome of the jewel beetle, Anthaxia chinensis (Coleoptera: Buprestidae). Mitochondrial DNA Part B Resources 6(10): 2962–2963.
  • Cobos A (1980) Ensayo sobre los géneros de la subfamilia Polycestinae (Coleoptera, Buprestidae) (Parte I). EOS Revista Española de Entomologia 54(1–4): 15–94.
  • Cobos A (1986) Fauna Iberica de Coleopteros Buprestidae. Consejo Superior de Invertigaciones Cientifificas, Madrid, 364 pp.
  • Dowton M, Castro LR, Austin AD (2002) Mitochondrial gene rearrangements as phylogenetic characters in the invertebrates: The examination of genome ‘morphology’. Invertebrate Systematics 16(3): 345–356.
  • Duan J, Quan GX, Mittapalli O, Cusson M, Krell PJ, Doucet D (2017) The complete mitogenome of the Emerald Ash Borer (EAB), Agrilus planipennis (Insecta: Coleoptera: Buprestidae). Mitochondrial DNA Part B Resources 2(1): 134–135.
  • Evans AM, Mckenna DD, Bellamy CL, Farrell BD (2015) Large-scale molecular phylogeny of metallic wood-boring beetles (Coleoptera: Buprestoidea) provides new insights into relationships and reveals multiple evolutionary origins of the larval leaf-mining habit. Systematic Entomology 40(2): 385–400.
  • Fenn JD, Cameron SL, Whiting RLF (2007) The complete mitochondrial genome sequence of the Mormon cricket (Anabrus simplex: Tetdgonildae: Orthoptera) and an analysis of control region variability. Insect Molecular Biology 16(2): 239–252.
  • Friedrich M, Muquim N (2003) Sequence and phylogenetic analysis of the complete mitochondrial genome of the flour beetle Tribolium castanaeum. Molecular Phylogenetics and Evolution 26(3): 502–512.
  • Greiner S, Lehwark P, Bock R (2019) OrganellarGenomeDRAW (OGDRAW) version 1.3.1: Expanded toolkit for the graphical visualization of organellar genomes. Nucleic Acids Research 47(W1): W59–W64.
  • Guindon S, Dufayard J, Lefort V, Anisimova M, Hordijk W, Gascuel O (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML 3.0. Systematic Biology 59(3): 307–321.
  • Hołyński RB (1988) Remarks on the general classification of Buprestidae Leach as applied to Maoraxiina. Folia Entomologica Hungarica 49(1): 49–54.
  • Hołyński RB (1993) A reassessment of the internal classification of the Buprestidae Leach (Coleoptera). Crystal. Series Zoologica [Göd] 1: 1–42.
  • Hołyński RB (2009) Taxonomic Structure of the Subtribe Chrysochroina Cast. with Review of the Genus Chrysochroa Dej. Gondwana, Warszawa, 391 pp.
  • Hołyński RB (2014) Review of the Indo-Pacific Buprestidae Leach (Coleoptera) I: Julodinae Lac. Gondwana, Warszawa, 85 pp.
  • Hong MY, Jeong HC, Kim MJ, Jeong HU, Lee SH, Kim I (2009) Complete mitogenome sequence of the jewel beetle, Chrysochroa fulgidissima (Coleoptera: Buprestidae). Mitochondrial DNA Mapping, Sequencing, and Analysis 20(2–3): 46–60.
  • Huang XY, Chen B, Wei ZH, Shi AM (2022) First report of complete mitochondrial genome in the tribes Coomaniellini and Dicercini (Coleoptera: Buprestidae) and phylogenetic implications. Genes 13(6): e1074.
  • Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A, Jermiin LS (2017) ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589.
  • Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28(12): 1647–1649.
  • Kolibáè J (2000) Classification and phylogeny of the Buprestoidea (Insecta: Coleoptera). Acta Musei Moraviae, Scientiae biologicae [Brno] 85: 113–184.
  • Krzywinski J, Li C, Morris M, Conn JE, Lima JB, Povoa MM, Wilkerson RC (2011) Analysis of the evolutionary forces shaping mitochondrial genomes of a Neotropical malaria vector complex. Molecular Phylogenetics and Evolution 58(3): 469–477.
  • Kubáň V, Volkovitsh MG, Kalashian MJ, Jendek E (2016) Family Buprestidae Leach, 1815. In: Löbl I, Löbl D (Eds) Catalogue of Palaearctic Coleoptera. Scarabaeoidea, Scirtoidea, Dascilloidea, Buprestoidea, Byrrhoidea. Revised and Updated Edition. Apollo Books, Stenstrup, 432–574.
  • Kück P, Meid SA, Groß C, Wägele JW, Misof B (2014) AliGROOVE–visualization of heterogeneous sequence divergence within multiple sequence alignments and detection of inflated branch support. Bioinformatics 15(1): e294.
  • Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33(7): 1870–1874.
  • Li H, Shao RF, Song N, Song F, Jiang P, Li ZH, Cai WZ (2015) Higher-level phylogeny of paraneopteran insects inferred from mitochondrial genome sequences. Scientific Reports 5(1): e8527.
  • Li R, Shu XH, Li XD, Meng L, Li BP (2019) Comparative mitogenome analysis of three species and monophyletic inference of Catantopinae (Orthoptera: Acridoidea). Genomics 111(6): 1728–1735.
  • Lim LWK, Chung HH, Lau MMLL, Aziz F, Gan HM (2021) Improving the phylogenetic resolution of Malaysian and Javan mahseer (Cyprinidae), Tor tambroides and Tor tambra: Whole mitogenomes sequencing, phylogeny and potential mitogenome markers. Gene 791: e145708.
  • Lowe TM, Chan PP (2016) tRNAscan-SE On-line: Integrating search and context for analysis of transfer RNA genes. Nucleic Acids Research 44(W1): W54–W57.
  • Motyka M, Kusy D, Háva J, Jahodářová E, Bílková R, Vogler AP, Bocak L (2022) Mitogenomic data elucidate the phylogeny and evolution of life strategies in Dermestidae (Coleoptera). Systematic Entomology 47(1): 82–93.
  • Nie RE, Wei J, Zhang SK, Vogler AP, Wu L, Konstantinov AS, Li WZ, Yang XK, Xue HJ (2019) Diversification of mitogenomes in three sympatric Altica flea beetles (Insecta, Chrysomelidae). Zoologica Scripta 48(5): 657–666.
  • Nie RE, Andújar C, Gómez-Rodríguez C, Bai M, Xue HJ, Tang M, Yang CT, Tang P, Kang XK, Vogler AP (2020) The phylogeny of leaf beetles (Chrysomelidae) inferred from mitochondrial genomes. Systematic Entomology 45(1): 188–204.
  • Nie R, Vogler AP, Yang XK, Lin MY (2021) Higher-level phylogeny of longhorn beetles (Coleoptera: Chrysomeloidea) inferred from mitochondrial genomes. Systematic Entomology 46(1): 56–70.
  • Park JS, Cho Y, Kim MJ, Nam SH, Kim I (2012) Description of complete mitochondrial genome of the black-veined white, Aporia crataegi (Lepidoptera: Papilionoidea), and comparison to papilionoid species. Journal of Asia-Pacific Entomology 15(3): 331–341.
  • Peng XJ, Liu J, Wang Z, Zhan QZ (2021) The complete mitochondrial genome of the pyrophilous jewel beetle Melanophila acuminata (Coleoptera: Buprestidae). Mitochondrial DNA Part B Resources 6(3): 1059–1060.
  • Perna NT, Kocher TD (1995) Patterns of nucleotide composition at fourfold degenerate sites of animal mitochondrial genomes. Journal of Molecular Evolution 41(3): 353–358.
  • Qin J, Zhang YZ, Zhou X, Kong XB, Wei SJ, Ward RD, Zhang AB (2015) Mitochondrial phylogenomics and genetic relationships of closely related pine moth (Lasiocampidae: Dendrolimus) species in China, using whole mitochondrial genomes. Genomics 16(1): 428.
  • Ronquist F, Teslenko M, Der Mark PV, Ayres DL, Darling AE, Hohna S, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice across a Large Model Space. Systematic Biology 61(3): 539–542.
  • Sheffield NC, Song H, Cameron SL, Whiting MF (2009) Nonstationary evolution and compositional heterogeneity in beetle mitochondrial phylogenomics. Systematic Biology 58(4): 381–394.
  • Sun HQ, Zhao WX, Lin RZ, Zhou ZF, Huai WX, Yao YX (2020) The conserved mitochondrial genome of the jewel beetle (Coleoptera: Buprestidae) and its phylogenetic implications for the suborder Polyphaga. Genomics 112(5): 3713–3721.
  • Thompson JD, Higgins DG, Gibson TJ (1994) Clustal W: Improving the sensitivity of progressive multiple sequence alignment through sequence weight, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22(22): 4673–4680.
  • Timmermans MJTN, Dodsworth S, Culverwell CL, Bocak L, Ahrens D, Littlewood DTJ, Pons J, Vogler AP (2010) Why barcode? High-throughput multiplex sequencing of mitochondrial genomes for molecular systematics. Nucleic Acids Research 38(21): e197.
  • Timmermans MJTN, Barton C, Haran J, Ahrens D, Culverwell CL, Ollikainen A, Dodsworth S, Foster PG, Bocak L, Vogler AP (2016) Family-level sampling of mitochondrial genomes in Coleoptera: Compositional heterogeneity and phylogenetics. Genome Biology and Evolution 8(1): 161–175.
  • Tôyama M (1987) The systematic positions of some buprestid genera (Coleoptera, Buprestidae). Elytra 15: 1–11.
  • Volkovitsh MG (2001) The comparative morphology of antennal structures in Buprestidae (Coleoptera): evolutionary trends, taxonomic and phylogenetic implications. Part 1. Acta Musei Moraviae, Scientiae Biologicae [Bron] 86: 43–169.
  • Wang X, Zhang H, Kitching I, Xu ZB, Huang YX (2021) First mitogenome of subfamily Langiinae (Lepidoptera: Sphingidae) with its phylogenetic implications. Gene 789: e145667.
  • Weng MQ, Wang Y, Huang J, Huang LL, Lin YQ, Zheng QL, Wu YZ, Wu SQ (2022) The complete mitochondrial genome of Chalcophora japonica chinensis Schaufuss, 1879 (Coleoptera: Buprestidae). Mitochondrial DNA Part B Resources 7(8): 1571–1573.
  • Wu C, Zhou Y, Tian T, Li TJ, Chen B (2022) First report of complete mitochondrial genome in the subfamily Alleculinae and mitochondrial genome-based phylogenetics in Tenebrionidae (Coleoptera: Tenebrionoidea). Insect Science 29(4): 1226–1238.
  • Xiao JH, Jia JG, Murphy RW, Huang DW (2011) Rapid evolution of the mitochondrial genome in chalcidoid wasps (Hymenoptera: Chalcidoidea) driven by parasitic lifestyles. PLoS ONE 6(11): e26645.
  • Xiao LF, Zhang SD, Long CP, Guo QY, Xu JS, Dai XH, Wang JG (2019) Complete mitogenome of a leaf-Mining buprestid Beetle, Trachys auricollis, and its phylogenetic implications. Genes 10(12): e992.
  • Yan L, Zhang M, Gao Y, Pape T, Zhang D (2017) First mitogenome for the subfamily Miltogramminae (Diptera: Sarcophagidae) and its phylogenetic implications. European Journal of Entomology 114(1): 422–429.
  • Yu P, Cheng X, Ma Y, Yu D, Zhang J (2016) The complete mitochondrial genome of Brachythemis contaminata (Odonata: Libellulidae). Mitochondrial DNA A DNA Mapping. Seqencing, and Analysis 27(3): 2272–2273.
  • Zhang DX, Hewitt GM (1997) Insect mitochondrial control region: A review of its structure, evolution and usefulness in evolutionary studies. Biochemical Systematics and Ecology 25(2): 99–120.
  • Zhang DX, Szymura JM, Hewitt GM (1995) Evolution and structural conservation of the control region of insect mitochondrial DNA. Journal of Molecular Evolution 40(4): 382–391.
  • Zhang D, Gao F, Jakovlić I, Zou H, Zhang J, Li WX, Wang GT (2020) PhyloSuite: An integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies. Molecular Ecology Resources 20(1): 348–355.
  • Zhang H, Lu CC, Liu Q, Zou TM, Qiao GX, Huang XL (2022) Insights into the evolution of aphid mitogenome features from new data and comparative analysis. Animals 12(5): e1970.
  • Zheng BY, Han YY, Yuan RZ, Liu JX, Achterberg C, Tang P, Chen XX (2022) Comparative mitochondrial genomics of 104 Darwin wasps (Hymenoptera: Ichneumonidae) and its implication for phylogeny. Insects 13(2): e124.

Supplementary material

Supplementary material 1 

First mitochondrial genome of subfamily Julodinae (Coleoptera, Buprestidae) with its phylogenetic implications

Zhonghua Wei, Xuyan Huang, Aimin Shi

Data type: table, images (word document)

Explanation note: Nucleotide composition of three newly generated mitogenomes. Circular maps of mitogenomes for Julodis variolaris, Ptosima chinensis, and Chalcophora japonica. The predicted secondary cloverleaf structure for the tRNAs of Julodis variolaris (image S2), Ptosima chinensis (image S3) and Chalcophora japonica (image S4). Phylogenetic relationships of Buprestidae using ML analyses based on 13 PCGs + 2 rRNAs of mitogenomes; the values one branches are bootstrap.

This dataset is made available under the Open Database License ( The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (2.02 MB)
login to comment