Molecular diagnosis of 5 silkworm strains endemic to South Korea using single-nucleotide polymorphisms selected from whole-genome sequences.

The domesticated silkworm, Bombyx mori Linnaeus (Lepidoptera: Bombycidae), often poses a challenge in strain identification due to similarities in morphology and genetic background. In South Korea, around 40 silkworm strains are classified as premium, including 5 endemic tri-molting strains: Goryeosammyeon, Sammyeonhonghoeback, Hansammyeon, Sun7ho, and Sandongsammyeon. These strains have potential for breeding programs in response to emerging industry demands, necessitating a reliable strain identification method. In this study, we established a molecular diagnosis approach for these 5 strains. We selected 2-4 single-nucleotide polymorphisms (SNPs) for each strain from whole-genome sequences of 39 strains, encompassing 37 previously studied and 2 newly added. These SNPs were utilized to construct decision trees for each endemic strain identification. The SNPs can be used to distinguish each target strain from the 38 nontarget strains by the tetra-primer amplification refractory mutation system-polymerase chain reaction, with the exception of HMS which needs the addition of PCR-restriction fragment length polymorphism method at the final step. This decision tree-based method using genomic SNPs, coupled with the 2 typing methods, produced consistent and accurate results, providing 100% accuracy. Additionally, the significant number of remaining SNPs identified in this study could be valuable for future diagnosis of the other strains.

Identification of Sympetrum depressiusculum Sélys, 1841 in South Korea (Odonata: Libellulidae) According to Morphology and Genetic Markers.

In South Korea, both Sympetrum depressiusculum Sélys, 1841 (Odonata: Libellulidae), which is distributed throughout Europe and from Russia to the Korean Peninsula, and Sympetrum frequens Sélys, 1883, which is endemic to Japan, are recorded. However, the identity of South Korean populations and the validity of listing the two species have not yet been settled. In this study, we collected seventy-four individuals of Sympetrum species from South Korea (five localities), Russia, The Netherlands, and Japan. These samples were examined for morphology and sequenced for partial COI, 16S rRNA, and a nuclear internal spacer (ITS) region, after which these molecular data were combined with available public data from Russia, Japan, and The Netherlands. Major morphological characters that have been used to distinguish the two species and phylogenetic, network, and structure analyses all consistently suggest that South Korean populations form a single species. Consequently, it could be valid to treat South Korean populations as one species, S. depressiusculum, by applying the senior name. Nevertheless, the validity of maintaining each as an independent species in other countries may need additional study considering that our samples were focused more on South Korea and limited for Europe, Russia, and Japan.

Reversible Na Plating/Stripping with High Areal Capacity Using an Electroconductive Liquid Electrolyte System.

Anode-free sodium-metal batteries (AFSMBs) are promising candidates for maximizing energy density and minimizing cost and safety hazards in the absence of metallic sodium during cell assembly. The practical implementation of AFSMBs is hindered by the low cycling stability of Na-metal plating and stripping, particularly under high areal capacities, due to unstable solid electrolyte interphase (SEI) layer formation with electrolyte decomposition and inactive dead Na formation. Here, we proposed an electroconductive electrolyte system consisting of liquid electrolytes that accept electrons at a certain energy level and form electronically conductive and solid electrolytes that prevent internal short circuit through low electronic conductivity. The electron acceptability and high electronic conductivity of the liquid electrolyte can suppress the irreversible electron transfer with electrolyte decomposition and reutilize the inactive dead metal, respectively. The functions of the system were demonstrated using a sodium biphenyl liquid electrolyte-NASICON solid electrolyte in a seawater battery (SWB) system, which features an infinite sodium source. The anode-free SWB cells achieved a high Coulombic efficiency of ≥99.9% for over 60 cycles at a high areal capacity of ∼24 mAh/cm2. This study provides insight into the Na plating/stripping properties in anode-free systems and proposes a significant strategy for improving the reversibility of metal anodes for various battery systems with solid electrolytes.

Correction: Oh et al. Enhanced Effect of Polyethyleneimine-Modified Graphene Oxide and Simvastatin on Osteogenic Differentiation of Murine Bone Marrow-Derived Mesenchymal Stem Cells. Biomedicines 2021, 9, 501.

Jeong-Sun Park was not included as an author in the original publication [...].

Complete mitochondrial genome of Asiagomphus coreanus (Odonata: Gomphidae), which is endemic to South Korea.

Asiagomphus coreanus (Doi & Okumura, 1937) belongs to the family Gomphidae in the order Odonata, and has been listed as an endemic species in South Korea. Here, we assembled its complete mitochondrial genome (mitogenome) which is 15,649 base pairs (bp) in length. The A. coreanus mitogeneome consists of a typical set of genes [13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, and 22 transfer RNA (tRNA) genes] and one major non-coding A + T-rich region which is 846 bp long. The gene arrangement of the species was identical to that of commonly found in the majority of the insects. Phylogenetic analyses using the concatenated sequences of 13 PCGs and two rRNA genes of the representative odonate mitogenomes by Bayesian inference method revealed that A. coreanus belongs to the Gomphidae family with a strong nodal support (Bayesian posterior probabilities = 1). Unlike previous phylogenetic analyses (with regards to suborder relationships) the suborder Anisozygoptera-which was represented by a single species, Epiophlebia superstes-was placed as the sister to Zygoptera.

Whole-genome sequences of 37 breeding line Bombyx mori strains and their phenotypes established since 1960s.

Bombyx mori is a key insect in the sericulture industry and one of the very important economic animals that are responsible for not only the livelihood of many farmers internationally but also expended biomedical use. The National Institute of Agricultural Sciences of the Rural Development Administration of Korea (NIAS, RDA, Korea) has been collecting silkworm resources with various phenotypic traits from the 1960s and established breeding lines for using them as genetic resources. And these breeding line strains have been used to develop suitable F1 hybrid strains for specific use. In this study, we report the whole-genome sequences of 37 breeding line B. mori strains established over the past 60 years, along with the description of their phenotypic characteristics with photos of developmental stages. In addition, we report the example phenotypic characteristics of the F1-hybrid strain using these breeding line strains. We hope this data will be used as valuable resources to the related research community for studying B. mori and similar other insects.

Phylogeographic Relationships among Bombyx mandarina (Lepidoptera: Bombycidae) Populations and Their Relationships to B. mori Inferred from Mitochondrial Genomes.

We report 37 mitochondrial genome (mitogenome) sequences of Bombyx mori strains (Lepidoptera: Bombycidae) and four of B. mandarina individuals, each preserved and collected, respectively, in South Korea. These mitogenome sequences combined with 45 public data showed a substantial genetic reduction in B. mori strains compared to the presumed ancestor B. mandarina, with the highest diversity detected in the Chinese origin B. mori. Chinese B. mandarina were divided into northern and southern groups, concordant to the Qinling-Huaihe line, and the northern group was placed as an immediate progenitor of monophyletic B. mori strains in phylogenetic analyses, as has previously been detected. However, one individual that was in close proximity to the south Qinling-Huaihe line was exceptional, belonging to the northern group. The enigmatic South Korean population of B. mandarina, which has often been regarded as a closer genetic group to Japan, was most similar to the northern Chinese group, evidencing substantial gene flow between the two regions. Although a substantial genetic divergence is present between B. mandarina in southern China and Japan, a highly supported sister relationship between the two regional populations may suggest the potential origin of Japanese B. mandarina from southern China instead of the Korean peninsula.

The first complete mitochondrial genome in the family Attevidae (Attevaaurea) of the order Lepidoptera.

The superfamily Yponomeutoidea, one of the early-derived groups in the order Lepidoptera, consists of 11 families. However, mitochondrial genome (mitogenome) sequences, popularly used for phylogeny and evolutionary tracing, are available for only seven species across six genera and five families. Thus, a larger variety of mitogenome sequences in Yponomeutoidea are required to improve our understanding of lepidopteran phylogeny and genomic evolution. In this study, we present the complete mitogenome of Attevaaurea (Fitch, 1856), the first species in the family Attevidae (superfamily Yponomeutoidea, order Lepidoptera) to be sequenced. The complete mitogenome comprises 16,329 bp and contains a typical set of genes and one non-coding region. Within Yponomeutoidea, the mitogenome of A.aurea has a unique trnI-trnM-trnQ arrangement at the A + T-rich region and ND2 junction and trnA-ND3 arrangement at the trnG and trnR junction. Twelve of the 13 protein-coding genes (PCGs) of A.aurea have a typical ATN starting codon, whereas COI has the atypical CGA codon, which is frequently found in the starting region of lepidopteran COI. Phylogenetic analyses, based on the concatenated sequences of 13 PCGs and two rRNA genes, using the Maximum Likelihood method, revealed a sister relationship between Attevidae and Praydidae with moderately low nodal support (bootstrap support = 64%).

Complete mitochondrial genome of Ricania shantungensis Chou & Lu, 1977 (Hemiptera: Ricaniidae).

Ricania shantungensis Chou & Lu, 1977 (Hemiptera: Ricaniidae), is an invasive pest that attacks forest as well as agricultural trees. We sequenced the 15,358 bp long complete mitochondrial genome (mitogenome) of this species; it consists of a typical set of genes (13 protein-coding genes, 2 rRNA genes, and 22 tRNA genes) and one major non-coding AT-rich region. The orientation and gene order of the R. shantungensis mitogenome are identical to that of the ancestral type found in majority of the insects. Bayesian inference (BI) phylogeny placed the R. shantungensis examined in our study, together with Ricania spp. in a group with the highest nodal support, forming the family Ricaniidae to which R. shantungensis belongs.

Complete Mitochondrial Genomes of Metcalfa pruinosa and Salurnis marginella (Hemiptera: Flatidae): Genomic Comparison and Phylogenetic Inference in Fulgoroidea.

The complete mitochondrial genomes (mitogenomes) of two DNA barcode-defined haplotypes of Metcalfa pruinosa and one of Salurnis marginella (Hemiptera: Flatidae) were sequenced and compared to those of other Fulgoroidea species. Furthermore, the mitogenome sequences were used to reconstruct phylogenetic relationships among fulgoroid families. The three mitogenomes, including that of the available species of Flatidae, commonly possessed distinctive structures in the 1702-1836 bp A+T-rich region, such as two repeat regions at each end and a large centered nonrepeat region. All members of the superfamily Fulgoroidea, including the Flatidae, consistently possessed a motiflike sequence (TAGTA) at the ND1 and trnS2 junction. The phylogenetic analyses consistently recovered the familial relationships of (((((Ricaniidae + Issidae) + Flatidae) + Fulgoridae) + Achilidae) + Derbidae) in the amino acid-based analysis, with the placement of Cixiidae and Delphacidae as the earliest-derived lineages of fulgoroid families, whereas the monophyly of Delphacidae was not congruent between tree-constructing algorithms.

Complete mitochondrial genome of Coleophora therinella Tengström, 1848 (Lepidoptera: Coleophoridae).

The mitochondrial genome (mitogenome) of Coleophora therinella Tengström, 1848 is the first report for the family Coleophoridae in Lepidoptera. The 15,539-bp long complete genome has an arrangement identical to that observed in most lepidopteran genomes. COI had the atypical CGA codon that is frequently found in the start region of the lepidopteran COI, and COII had the GTG codon found previously in Drosophila yakuba ND5 and Rattus norvegicus ND1. The 457-bp long A + T-rich region was the second largest, next to Blastobasis lacticolella, which belongs to Blastobasidae in the superfamily Gelechioidea. The A/T content of the whole mitogenome was 80.7%; however, it varied among the regions/genes as follows: A + T-rich region, 94.8%; srRNA, 85.0%; lrRNA, 84.3%; tRNAs, 81.5%; and PCGs, 78.9%. Phylogenetic analyses with concatenated sequences of the 13 PCGs and two RNA genes using the maximum likelihood method, placed Coleophoridae, represented only by C. therinella, as the most basal lineage of the Gelechioidea families consisted of Stathmopodidae, Scythrididae, Blastobasidae, Autostichidae, and Oecophoridae, but nodal support for this grouping was very low (27%). Currently, several families of Gelechioidea are represented by a single species. Thus, extended sampling is required for further reasonable inference for the relationships of these families.

The complete mitochondrial genome of Monopis longella Walker, 1863 (Lepidoptera: Tineidae).

The complete mitochondrial genome (mitogenome) of Monopis longella Walker, 1863 (Lepidoptera: Tineidae) comprises 15,541 bp and contains a typical set of genes and one non-coding region. The gene arrangement of M. longella is unique for Lepidoptera in that it has a trnI-trnM-trnQ sequence in the A + T-rich region and ND2 junction. Unlike most other lepidopteran insects, in which the COI gene has CGA as the start codon, M. longella COI has an ATT codon. Phylogenetic analyses based on the concatenated sequences of 13 protein-coding genes and two rRNA genes, using the Bayesian inference (BI) method, placed M. longella in the Tineidae, sister in position to the cofamilial species, Tineola bisselliella, with the highest nodal support. Tineidae, represented by three species including M. longella, formed a monophyletic group with high support (Bayesian posterior probability = 0.99). Within Tineoidea the sister relationship between Tineidae and Meessiidae was obtained with the highest support, leaving Psychidae occupying the basal lineage of the two families.

Complete mitochondrial genome of the highly fecund Bombyx mori linnaeus, 1758 (Lepidoptera: Bombycidae) strain Jam 146.

To meet the increasing demands of the society in the current era, new strains of the domesticated silkworm Bombyx mori Linnaeus, 1758 (Lepidoptera: Bombycidae) are being continuously bred. Consequently, cataloging the genetic information of pure lines is essential. The strain Jam 146, whose larvae have atypical pale, crescent-shaped body markings, is an important breeding resource due to its excellent fecundity. In this study, we sequenced the mitochondrial genome (mitogenome) of this strain using next-generation sequencing. The complete genome of this strain has a gene arrangement typical of Lepidoptera. The length of the Jam 146 mitogenome (15,661 bp) is well within the range reported in other B. mori strains, i.e. between 15,629 (Baiyun strain, China) and 15,676 bp (Hukpyobeom strain, South Korea). However, the total length of protein-coding genes, 3,733 codons in Jam 146 and two other silkworm strains previously reported from South Korea, is 13 codons longer than that in other B. mori strains. Phylogenetic analysis of 22 silkworm strains from nine countries showed that the Jam 146 strain forms a strong cluster with three other strains from China, Japan, and South Korea, suggesting that after their split from a common ancestor, the evolutionary divergence among the silkworm strains in these countries has been limited.

Enhanced Effect of Polyethyleneimine-Modified Graphene Oxide and Simvastatin on Osteogenic Differentiation of Murine Bone Marrow-Derived Mesenchymal Stem Cells.

Statin derivatives traditionally have been used for the treatment of hyperlipidemia, but recent studies have shown their ability to regulate bone metabolism and promote bone growth. In this study, simvastatin (Sim), a new therapeutic candidate for bone regeneration, was combined with graphene oxide (GO), which has recently attracted much interest as a drug delivery method, to produce a compound substance effective for bone regeneration. To create a stable and homogenous complex with Sim, GO was modified with polyethylenimine, and the effect of modification was analyzed using Fourier transform infrared spectroscopy, zeta potential, and cytotoxicity testing. More specifically, the osteogenic differentiation potential expected by the combination of the two effective materials for osteogenic differentiation, GO and Sim, was evaluated in mesenchymal stem cells. Compared with control groups with GO and Sim used separately, the GO/Sim complex showed excellent osteogenic differentiation properties, with especially enhanced effects in the complex containing < 1 µM Sim.

Complete mitochondrial genome of the summer heath fritillary butterfly, Mellicta ambigua (Lepidoptera: Nymphalidae).

We sequenced the mitochondrial genome (mitogeome) of the summer heath fritillary bullterfly, Mellicta ambigua Ménétriès, 1859 (Lepidoptera: Nymphalidae), which is listed as an endangered insect in South Korea. The 15,205-bp long complete genome contained 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes, and 1 A + T-rich region with an arrangement identical to that observed in most insect mitogenomes. Unlike the other PCGs, COI had the atypical CGA start codon frequently found in lepidopteran COI. The A/T content of the whole mitogenome was 80.57%; however, it varied among the regions/genes as follows: A + T-rich region, 93.39%; srRNA, 85.37%; lrRNA, 84.92%; tRNAs, 81.13%; and PCGs, 79.22%. Phylogenetic analyses using concatenated sequences of the 13 PCGs and 2 rRNAs placed M. ambigua as a sister group to the within-tribe species, Melitaea cinxia, with the highest nodal support both in the maximum-likelihood (ML) and Bayesian inference (BI) methods.

Complete mitochondrial genome of Acanthopsyche nigraplaga (Lepidoptera: Psychidae).

We report the mitochondrial genome (mitogenome) of a case-making moth Acanthopsyche nigraplaga Wileman, 1911 (Lepidoptera: Psychidae). The 15,704 bp long complete mitogenome comprises a typical set of genes [13 protein-coding genes (PCGs), 2 rRNA genes, and 22 tRNA genes] and one major non-coding, A + T-rich region, with an arrangement identical to that observed in most lepidopteran mitogenomes. Twelve of the 13 PCGs of the A. nigraplaga mitogenome initiate with a typical ATN start codon, however COI contains the atypical CGA start codon that is common for lepidopteran COI genes. A phylogenetic analysis using concatenated nucleotide sequences of the 13 PCGs and 2 rRNA genes using the Bayesian inference method fully resolved A. nigraplaga in a monophyletic clade within the Psychidae. Acanthopsyche nigraplaga was situated in a sister position to Eumeta variegata and Mahasena oolona with high nodal support. As more mitogenome sequences are available further scrutinized analysis for the superfamily Tineoidea including Psychidae will be possible.

Complete mitochondrial genome of Pterodecta felderi (Lepidoptera: Callidulidae).

We report the mitochondrial genome (mitogenome) of Pterodecta felderi (Callidulidae: Lepidoptera), which is the first mitogenome sequences in the family Callidulidae, a monotypic family in the superfamily Calliduloidea. The 15,340-bp long complete mitogenome consists of a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and 1 major non-coding A + T-rich region, which are arranged in a way that is frequently observed in Lepidoptera. Of the 13 PCGs, 12 P. felderi start with ATN, except for COI, which starts with CGA. The P. felderi mitogenome consists of 210-bp long intergenic-spacer sequences and 27-bp long overlaps. Phylogenetic analysis of superfamilial relationships in the lepidopteran clade Obtectomera with concatenated sequences of the 13 PCGs and 2 rRNA genes using the Bayesian inference method showed that Calliduloidea, which is only represented by P. felderi, was placed as the most basal lineage about Macroheterocera (Lasiocampoidea, Bombycoidea, Mimallonoidea, Noctuoidea, and Drepanoidea), Papilionoidea, and Pyraloidea.

Mitochondrial genome of the mason bee, Osmia pedicornis (Hymenopetra: Megachilidae).

The mason bee, Osmia pedicornis Cockerell, 1919, which is importantly used as the pollinator, particularly for apples in Korea. We sequenced the mitochondrial genome (mitogenome) of O. pedicornis as an initial study for species identification and subsequent population genetic study. The size of the incomplete genome was 14,505 bp, excluding the trnA, trnC, and the A + T-rich region that were unable to sequence, but including partially sequenced trnM and srRNA. The genome included typical sets of protein-coding genes (PCGs), rRNA genes, and one non-coding region, tRNAs, excluding two unidentified tRNAs. Although positions of the two tRNAs that were not sequenced are unknown the gene arrangement of O. pedicornis mitogenome has the tRNA arrangement, trnM-trnQ-trnI, at the A + T-rich region and ND2 junction that differed from that of previously published O. excavate, which has trnA-trnQ-trnI arrangement at the junction. Phylogenetic analyses were performed using concatenated sequences of the 13 PCGs genes and the maximum likelihood method with the inclusion of a total of 12 mitogenome sequences belonging to three families in the superfamily Apoidea. Current O. pedicornis was placed as the sister to the O. bicornis, with the highest nodal support. The Apidae and Megachilidae were placed as the sister group, with the placement of Colletidae as the basal lineage for the group with the highest nodal support.

Tracing the Invasion and Expansion Characteristics of the Flatid Planthopper, Metcalfa pruinosa (Hemiptera: Flatidae), in Korea Using Mitochondrial DNA Sequences.

The flatid planthopper, Metcalfa pruinosa (Hemiptera: Flatidae), which is an invasive species, is widespread in Korea. We sequenced a fragment of the COI from 536 individuals collected mainly in Korea and the European countries and combined these sequence data with the public data, totaling 830 individuals worldwide. The identification of one shared haplotype only between Korea and the USA, the presence of this haplotype only in the North-West region of Korea, and the highest haplotype diversity in this region suggested that the North-West region is another point of entry in addition to the South-East region, which is the presumed sole point of entry to Korea. Furthermore, it suggested that North-West entry involves the M. pruinosa originating from the USA. In an effort to find further variable regions in the mitochondrial genome, one region provided substantially increased variability compared to that of the fragment of COI. F ST estimation, PCoA, and BAPS analysis, using the concatenated sequences of COI and the newly detected variable region to infer the expansion pattern in Korea, indicates that the main highway, running obliquely between the North-West and South-East regions, appears to be responsible for the current population genetic structure of M. pruinosa in Korea, facilitating gene flow through this highway traffic.

Complete mitochondrial genome of Scythris sinensis (Lepidoptera: Scythrididae).

The complete mitochondrial genome (mitogenome) of Scythris sinensis Felder & Rogenhofer, 1775 (Lepidoptera: Scythrididae) was 15,216 bp with a typical set of genes (13 protein-coding genes [PCGs], 2 rRNA genes, and 22 tRNA genes) and one non-coding region, with an arrangement identical to that observed in most lepidopteran genomes. Twelve PCGs had the typical ATN start codon, whereas COI had the atypical CGA codon that is frequently found in the start region of the lepidopteran COI. The 271-bp long A + T-rich region was the shortest among sequenced Gelechioidea, which ranged from 290 - 375 bp. Phylogenetic analyses with concatenated sequences of the 13 PCGs, two RNA genes, and 22 tRNA genes using the Bayesian inference (BI) method, placed S. sinensis in the Scythrididae, as a sister to the family Stathmopodidae. The nodal support for this sister relationship was the highest at Bayesian posterior probabilities = 1.