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. 2021 Mar 23;6(3):1144–1146. doi: 10.1080/23802359.2021.1901617

The complete mitochondrial genome of the two-spotted cricket Gryllus bimaculatus (Orthoptera: Gryllidae) from South Korea

Bia Park a,*, Eun Hwa Choi a,*, Gyeongmin Kim a,b, Cho Rong Shin a,b, Jihye Hwang a, Su Youn Baek a, Ui Wook Hwang a,c,d,✉
PMCID: PMC7995845  PMID: 33796770

Abstract

The complete mitochondrial genome of a two-spotted cricket Gryllus bimaculatus (Orthoptera: Gryllidae) from South Korea is determined and characterized in this study. The circular genome is 16,075 bp long, which consists of 13 protein-coding genes (PCGs), 2 ribosomal RNA genes, 22 transfer RNA genes, and an A + T−rich region. It has a base composition of A (40.35%), G (9.09%), C (16.80%), and T (33.76%). The gene order is identical to the ancestral gene arrangement pattern generally shown in arthropods, with the exception of an inversion of trnN-trnS1-trnE into trnE-trnS1-trnN. The maximum likelihood (ML) tree supports that G. bimaculatus is a distinct member of the monophyletic family Gryllidae.

Keywords: Gryllidae, Gryllus bimaculatus, mitochondrial genome, molecular phylogeny

The two-spotted cricket Gryllus bimaculatus De Geer, 1773, one of the most abundant cricket species, is widely distributed in tropical and subtropical regions of Asia, Africa, and southern Europe (Ragge 1972; Harrison and Bogdanowicz 1995; Mito and Noji 2008). This species belongs to the family Gryllidae within the superfamily Grylloidea (Order Orthoptera), which is known as a general food ingredient in South Korea (i.e. Kim et al. 2020). Generally, G. bimaculatus can be easily distinguished from the other species belonging to the genus Gryllus by the two spots on the base of its wings. Recently, the complete mitochondrial genome of G. bimaculatus was reported from China (Wang et al. 2019). Here, we fully determined the mitochondrial genome of G. bimaculatus from South Korea, which was compared with that of Chinese one previously known, with examination of the phylogenetic position within the family Gryllidae.

In this study, a reared specimen of G. bimaculatus was obtained from National Institute of Agricultural Sciences, Wanju-gun 55365, South Korea (35°49′45.45″N, 127°02′27.13″E). The specimen is kept in the Kyungpook National University (KNU), Daegu 41566, South Korea (voucher no. KNU2020001) and the extracted genomic DNA is stored in the −80 °C in the same KNU depository. The genomic DNA from South Korean specimen was extracted from the whole body using a DNeasy Blood & Tissue kit (Qiagen, Hilden, Germany). The 150 bp paired-end reads were generated by Hiseq X Ten platform (Illumina, San Diego, CA) sequencing of libraries contacting inserts of ca. 520 bp. The mitochondrial genome was extracted from the whole-genome sequencing data (6.6 Gb read sequences, unpublished data) using the program DeconSeq version 0.4.3 (http://deconseq.sourceforge.net/). The raw data of mitochondrial genome were mapped and annotated using the assembler gsMapper version 2.8 (Roche Inc., Basel, Switzerland) with GenBank DB as the reference (e.g. G. bimaculatus [MK204367], Velarifictorus hemelytrus [NC030762], and Teleogryllus emma [NC011823]).

The mitochondrial genome of the South Korean G. bimaculatus was completely sequenced, which is 16,075 bp in length (GenBank accession no. MT993975), and contains a standard gene component set including 13 protein-coding genes (PCGs) (cox1-3, cytb, nad1-6, nad4L, atp6, and atp8), 2 ribosomal RNA genes (rrnL and rrnS), 22 transfer RNA genes, and a non-coding A + T−rich region (CR). Of the 37 typical mitochondrial genes, 21 position on the heavy strand (H-strand) and the remaining 16 on the light strand (L-strand). The overall genome components and gene order are identical to those of the Chinese G. bimaculatus published by Wang et al. (2019). In comparison with the ancestral arthropod gene order (trnN-trnS1-trnE on the H-strand) (Boore 1999; Woo et al. 2007; Ryu and Hwang 2010; Baek et al. 2014; Park et al. 2016), an inverted gene order (trnE-trnS1-trnN on the L-strand) is observed in G. bimaculatus from South Korea and China. The inverted feature is known to be commonly found in all sequenced mitochondrial genomes of the family Gryllidae (Ma and Li 2018). The base composition is 40.35% for A, 9.09% for G, 16.80% for C, 33.7% for T, respectively, which indicates that A + T content (74.11%) is apparently higher than G + C content (25.89%). In all PCGs, the five A + T-biased codons are mainly used: ATT, ATA, AAT, TTA, and AAA. The longest PCG is 1,734 bp for nad5, whereas the shortest 156 bp for atp8. The common start codon is ATT or ATG, except for cox1 with TCG and nad1 with TTG. TAA is the most frequent stop codon, except for the nad3 and nad4 with TAG. All tRNAs except trnS1 lacked a stable dihydrouridine (DHU) arm have the typical clover-leaf secondary structure, which is a common phenomenon, generally found in metazoan mitochondrial tRNAs (Wolstenholme 1992).

To elucidate the phylogenetic position of the two-spotted cricket G. bimaculatus within the family Gryllidae, the nucleotide sequences of 13 PCGs were aligned and analyzed from 20 orthopteran mitochondrial genomes, out of which Locusta migratoria (Acrididae) was employed as an outgroup. The phylogenetic analysis was conducted by a maximum likelihood (ML) method in IQ-TREE web server (Trifinopoulos et al. 2016) and the best fitting model GTR + F + I + G4 was selected by ModelFinder (Kalyaanamoorthy et al. 2017). As shown in Figure 1, the South Korean and Chinese specimens of G. bimaculatus are grouped together, placing within the monophyletic family Gryllidae. It is expected that this study could be helpful in elucidating genetic diversity between two-spotted crickets in East Asia including South Korea, China, Japan, etc., and moreover in making a conservation of such edible insect species to become potential food resources for the future.

Figure 1.

Figure 1.

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A maximum-likelihood tree reconstructed with the nucleotide sequences of 13 mitochondrial protein-coding genes (PCGs) showing relationships among 20 orthopteran species. It indicates the monophyly of the family Gryllidae, within which two specimens of G. bimaculatus from South Korea and China place together. Locusta migratoria (Acrididae) is used as an outgroup. Branch supports are inferred from the ultrafast bootstrap method using IQ-TREE web server (Trifinopoulos et al. 2016).

Funding Statement

This research was supported by the Commercialization Promotion Agency for R&D Outcomes (COMPA) grant funded by the Korea government (MSIT) [Grant No. 2020-0003 to UWH], and National R&D Program through the National Research Foundation of Korea (NRF) funded by Ministry of Science and ICT [2020R1A6A3A01095736 to BP and 2020R1A6A3A0109563 to JH].

Disclosure statement

The authors report no conflict of interest.

Data availability statement

The genome sequence data that support the findings of this study are openly available in GenBank of NCBI at (https://www.ncbi.nlm.nih.gov/) under the accession no. MT993975. The associated BioProject, SRA, and Bio-Sample numbers are PRJNA704187, SRR13773776, and SANB17487625, respectively.

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

The genome sequence data that support the findings of this study are openly available in GenBank of NCBI at (https://www.ncbi.nlm.nih.gov/) under the accession no. MT993975. The associated BioProject, SRA, and Bio-Sample numbers are PRJNA704187, SRR13773776, and SANB17487625, respectively.

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