Polyandry may mitigate the negative impact of reproductive interference among bumblebees in Japan.

In social hymenopterans, monandry of the queen is an ancestral trait, and polyandry is a derived trait. Polyandry of the queen is the norm in a limited number of lineages, such as honeybees, leaf-cutting ants, Pogonomyrmex ants, and Vespula wasps, which presumably provide fitness advantages for the whole colony. The queen of the introduced bumblebee, Bombus terrestris, is polyandrous in Japan, whereas it is monandrous in native regions. We hypothesize that polyandry can evolve in a process that avoids the negative impacts of reproductive interference caused by interspecific mating and conducted genetic studies of the invasive species B. terrestris and two native subspecies, Bombus hypocrita sapporoensis and Bombus hypocrita hypocrita, in Japan. Our results revealed that although the native queens of B. hypocrita hypocrita allopatric with B. terrestris were strictly monandrous, the native queens of B. hypocrita sapporoensis sympatric with B. terrestris were polyandrous. These results suggested that the queens of native B. hypocrita sapporoensis do not experience negative impacts on interspecific mating from the invasive B. terrestris. We discuss the possibility that reproductive interference is a driving force in selection for multiple mating through an arms race between sympatric species.

Genetic differentiation and local adaptation of the Japanese honeybee, Apis cerana japonica.

We examine the population genetic structure and divergence among the regional populations of the Japanese honeybee, Apis cerana japonica, by re-sequencing the genomes of 105 individuals from the three main Japanese islands with diverse climates. The genetic structure results indicated that these individuals are distinct from the mainland Chinese A. cerana samples. Furthermore, population structure analyses have identified three genetically distinct geographic regions in Japan: Northern (Tohoku-Kanto-Chubu districts), Central (Chugoku district), and Southern (Kyushu district). In some districts, "possible non-native" individuals, likely introduced from other regions in recent years, were discovered. Then, genome-wide scans were conducted to detect candidate genes for adaptation by two different approaches. We performed a population branch statistics (PBS) analysis to identify candidate genes for population-specific divergence. A latent factor mixed model (LFMM) was used to identify genes associated with climatic variables along a geographic gradient. The PBSmax analysis identified 25 candidate genes for population-specific divergence whereas the LFMM analysis identified 73 candidate genes for adaptation to climatic variables along a geographic gradient. However, no common genes were identified by both methods.

Cross-mating between the alien bumblebee Bombus terrestris and two native Japanese bumblebees, B. hypocrita sapporensis and B. cryptarum florilegus, in the Nemuro Peninsula, Japan.

The rapid naturalization of Bombus terrestris across the Nemuro Peninsula has led to a decline in two closely related native Japanese species, namely Bombus hypocrita sapporensis and Bombus cryptarum florilegus, both belonging to the common subgenus Bombus. Although it is widely believed that cross-mating of native and non-native species is influenced by the common male sex pheromone in this region, no study has been conducted to substantiate this claim. Thus, we investigated the cross-activities of male sex pheromones between native and non-native bumblebees, as well as the frequencies of cross-mating, using chemical and DNA assays. Our gas chromatography-electroantennographic detector analyses and behavioral tests revealed the presence of sex pheromonal cross-activities between B. terrestris and the two Japanese bumblebees species. Furthermore, DNA analyses revealed the occurrence of cross-mating between native and non-native species in the Nemuro Peninsula. Overall, these results indicate the immediate need for conservation measures to safeguard Japanese bumblebee populations in the Nemuro Peninsula.

Formation of Amino Acids and Carboxylic Acids in Weakly Reducing Planetary Atmospheres by Solar Energetic Particles from the Young Sun.

Life most likely started during the Hadean Eon; however, the environmental conditions which contributed to the complexity of its chemistry are poorly known. A better understanding of various environmental conditions, including global (heliospheric) and local (atmospheric, surface, and oceanic), along with the internal dynamic conditions of the early Earth, are required to understand the onset of abiogenesis. Herein, we examine the contributions of galactic cosmic rays (GCRs) and solar energetic particles (SEPs) associated with superflares from the young Sun to the formation of amino acids and carboxylic acids in weakly reduced gas mixtures representing the early Earth's atmosphere. We also compare the products with those introduced by lightning events and solar ultraviolet light (UV). In a series of laboratory experiments, we detected and characterized the formation of amino acids and carboxylic acids via proton irradiation of a mixture of carbon dioxide, methane, nitrogen, and water in various mixing ratios. These experiments show the detection of amino acids after acid hydrolysis when 0.5% (v/v) of initial methane was introduced to the gas mixture. In the set of experiments with spark discharges (simulation of lightning flashes) performed for the same gas mixture, we found that at least 15% methane was required to detect the formation of amino acids, and no amino acids were detected in experiments via UV irradiation, even when 50% methane was used. Carboxylic acids were formed in non-reducing gas mixtures (0% methane) by proton irradiation and spark discharges. Hence, we suggest that GCRs and SEP events from the young Sun represent the most effective energy sources for the prebiotic formation of biologically important organic compounds from weakly reducing atmospheres. Since the energy flux of space weather, which generated frequent SEPs from the young Sun in the first 600 million years after the birth of the solar system, was expected to be much greater than that of GCRs, we conclude that SEP-driven energetic protons are the most promising energy sources for the prebiotic production of bioorganic compounds in the atmosphere of the Hadean Earth.

Complete mitochondrial DNA sequence of the Eastern Asian catfish Silurus asotus (Siluriformes: Siluridae) from Lake Biwa in Japan.

The east Asian catfish Silurus asotus is a common species living in fresh water in Japan. The complete mitochondrial genome of the S. asotus from Lake Biwa in Japan was analyzed using next-generation sequencing. The mitochondrial genome of S. asotus was identified as a 16,515 bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes, along with one A + T-rich control region. The AT content was 56.1%. Start codons ATG and GTG were found in 13 PCGs. Stop codons TAA, TAG, and AGA were observed in 13 PCGs. The heavy (H)-strand was predicted to have 12 PCGs and 14 tRNA and two rRNA genes, while the light (L)-strand was predicted to contain one PCGs and eight tRNA genes. The molecular phylogenetic analysis showed that S. asotus from Lake Biwa is genetically similar to S. asotus from China.

Complete mitochondrial DNA sequence of the Japanese endemic catfish Silurus biwaensis (Siluriformes: Siluridae) from Lake Biwa.

The Japanese endemic catfish Silurus biwaensis is distributed only in Lake Biwa and Yodo river drainages. There are four species of the genus Silurus in Japan, of which S. biwaensis has a most limited distribution. This catfish needs to be collected for DNA data owing to the lack of information related to its phylogenetic relationship. Here, the complete mitochondrial genome of the S. biwaensis from Lake Biwa in Japan was analyzed using next-generation sequencing. The mitochondrial genome of S. biwaensis was identified as a 16,531 bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes, along with one A + T-rich control region. The AT content was 55.83%. The heavy (H)-strand was predicted to have 12 PCGs, 14 tRNA, and 2 rRNA genes, whereas the light (L)-strand was predicted to contain one PCG and eight tRNA genes. The start codons ATG, ATC, and GTG were found in 13 PCGs. The stop codons TAA, TAG, and AGA were observed in all PCGs, except CytB and COX3. All tRNA genes formed typical cloverleaf secondary structures. The molecular phylogenetic relationships estimated using 13 PCGs (maximum-likelihood method) indicated that S. biwaensis is genetically distinct from the sympatric species S. asotus and S. lithophilus. This result clearly indicated that S. biwaensis is a valid species.

Complete mitochondrial DNA sequence of the Japanese endemic catfish Silurus lithophilus (Siluriformes: Siluridae).

The Japanese endemic catfish Silurus lithophilus is distributed only in Lake Biwa, Lake Yogo, and their river drainages. There are four species of the genus Silurus in Japan, of which S. lithophilus has a limited distribution. This catfish needs to be collected for DNA data owing to the lack of information related to its conservation. Here, the complete mitochondrial genome of the S. lithophilus from Lake Biwa in Japan was analyzed using next-generation sequencing. The mitochondrial genome of S. lithophilus was identified as a 16,524 bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes, along with one A + T-rich control region. The AT content was 55.89%. The heavy (H)-strand was predicted to have 12 PCGs and 15 tRNA, and 2 rRNA genes, whereas the light (L)-strand was predicted to contain one PCG and seven tRNA genes. The start codons ATG and GTG were found in 13 PCGs. The stop codons TAA, TAG, and AGA were observed in all PCGs, except CytB. All tRNA genes formed typical cloverleaf secondary structures. The molecular phylogenetic relationship, inferred using 13 PCGs (based on the maximum likelihood), was consistent with that reported in previous studies, which predicted a sister relationship between S. lithophilus and S. asotus. The results also clearly indicated that S. lithophilus is a valid species.

Complete mitochondrial DNA sequence of the parasitic honey bee mite Varroa destructor (Mesostigmata: Varroidae).

Varroa destructor is a parasite mite of the eastern honey bee Apis cerana, which is native to Asia. The European honey bee Apis mellifera was imported to Asia from Europe and the USA for apiculture in the 19th century. In a short period of time, V. destructor parasitized the artificially introduced honey bees. Varroa destructor was estimated to have spread around the world with A. mellifera when it was exported from Asia to locations worldwide about 50 years ago. The mitochondrial DNA of the parasitic honey bee mite V. destructor was analyzed using next-generation sequencing. The complete mitochondrial genome of V. destructor was identified as a 16,476-bp circular molecule containing 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one AT-rich control region. The heavy strand was predicted to have nine PCGs and 13 tRNA genes, whereas the light strand was predicted to contain four PCGs, nine tRNA genes, and two rRNA genes. All PCGs began with ATA as the start codon, except COIII and CytB, which had ATG as the start codon. Stop codons were of two types: TAA for eight genes and TAG for five genes. Molecular phylogenetic analysis revealed that V. destructor from Japan was genetically distant from that of France. A high base substitution rate of 2.82% was also confirmed between the complete mitochondrial DNA sequences of V. destructor from Japan and the USA, suggesting that one Varroa mite strain found in the USA is not from Japan.

Complete mitochondrial DNA sequence of the giant water bug Kirkaldyia deyrolli (Hemiptera: Belostomatidae).

Giant water bugs (genus Kirkaldyia and Lethocerus) are well known species from aquatic habitats throughout the world's subtropical and tropical areas. Only one species of the Kirkaldyia deyrolli is distributed in Japan. K. deyrolli has been designated as second category rare species according Law for Conservation of Endangered Species of Wild Fauna and Flora in Japan, in 2019. We analyzed, for the first time, the complete mitochondrial genomes of the giant water bug K. deyrolli from Japan using next-generation sequencing. The mitochondrial genome was a circular and 15,579 bp molecule that included 13 protein-coding genes (PCGs), 22 tRNA genes, and two ribosomal RNA genes, along with one AT-rich control region. The AT content value was 69.55%. The heavy strand was predicted to have nine PCGs and 15 tRNA genes, whereas the light strand was predicted to contain four PCGs, seven tRNA genes, and two rRNA genes. Start codons were variable for all PCGs: four ATA, three ATC, four ATG, two ATT genes as the start codon. Stop codons were of two types: TAA for 12 genes and TAG for one gene. Incomplete stop codon T was identified. The molecular phylogenetic relationship, inferred using 13 PCGs, was consistent with that reported in previous studies, which predicted a sister relationship to the genus Lethocerus.

A revision of subspecies structure of western honey bee Apis mellifera.

The taxonomy of honey bee A. mellifera contains a lot of issues due to the specificity of population structure, features of biology and resolutions of honey bee subspecies discrimination methods. There are a lot of transition zones between ranges of subspecies which led to the gradual changes of characteristics among neighbor subspecies. The modern taxonomic pattern of honey bee Apis mellifera is given in this paper. Thirty-three distinct honey bee subspecies are distributed across all Africa (11 subspecies), Western Asia and the Middle East (9 subspecies), and Europe (13 subspecies). All honey bee subspecies are subdivided into 5 evolutionary lineages: lineage A (10 subspecies) and its sublineage Z (3 subspecies), lineage M (3 subspecies), lineage C (10 subspecies), lineage O (3 subspecies), lineage Y (1 subspecies), lineage C or O (3 subspecies).

Complete mitochondrial DNA sequence of the East Asian minnow, Pungtungia herzi (Actinopterygii: Cypriniformes).

Some populations of the East Asian minnow Pungtungia herzi Herzenstein, 1892, which are naturally distributed in western Japan, have declined and are considered endangered. However, P. herzi has become a domestic invasive species in eastern Japan. Thus, knowledge of genetic features and phylogenetic relationships of P. herzi is important for conservation of this species and understanding its impact on ecosystems. We analyzed the complete mitochondrial genome using next generation sequencing of the East Asian minnow P. herzi from Yodo River, Osaka Prefecture, Japan. The mitochondrial genome of P. herzi consists of a circular molecule of 16,599 bp that includes 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one control region. The heavy (H)-strand was predicted to have 12 PCGs, 14 tRNA, and two rRNA genes, while the light (L)-strand was predicted to contain one PCGs and eight tRNA genes. The average AT content was 57.68%. The genes ATP8 and ATP6, ATP6 and COIII, ND4L and ND4, and ND5 and ND6 shared seven, one, seven, and four nucleotides, respectively. The initiation codons ATG and GTG were found in 12 and one genes, respectively. The termination codons TAA, TAG, incomplete TA-, and single T-- were observed in nine, one, one, and two genes, respectively. All the tRNA genes possessed a cloverleaf secondary structure. The phylogenetic relationships inferred using 13 PCGs (based on the maximum likelihood) were consistent with previous studies that predicted interrelationships of Cypriniformes.

Complete mitochondrial DNA sequence of the paper wasp Polistes riparius (Hymenoptera: Vespidae).

The paper wasp Polistes riparius is distributed in cold regions of northern East Asia to Russia. P. riparius are characterized by having longer cells than those of the closely related P. chinensis, which has a similar life history, as an adaptation to cold regions. The phylogenetic relationships of paper wasps have recently been studied; however, the genetic diversity and population structure of P. riparius has not been determined. The present study is the first to analyze the complete mitochondrial genome using next generation sequencing of P. riparius collected from Sapporo, Hokkaido Prefecture, Japan. The genome consisted of a closed loop that was 16,383 bp-long and included 13 protein coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one AT-rich control region. The average AT content was 84.54%. The heavy (H)-strand was predicted to have 12 PCGs and 14 tRNA genes, while the light (L)-strand was predicted to contain one PCGs, eight tRNA genes, and two rRNA genes. All PCGs started with ATG. Stop codons were of two types: TAA for 11 genes (ND1, ND2, ND3, ND4L, ND5, ND6, COXI, COXII, COXIII, COB, ATP6 and ATP8) and TAG for two genes (ND3 and ND4). The molecular phylogenetic relationship based on the maximum likelihood method using 13 PCGs was consistent with some previous studies in which a closely relationship between P. riparius and P. jokahamae.

Reproductive interference by alien hornet Vespa velutina threatens the native populations of Vespa simillima in Japan.

The yellow-legged hornet Vespa velutina has become one of the major alien species in European and East Asian countries. As in its homeland, the invading V. velutina is reported as the major predator of honeybees and is becoming a threat to beekeeping in Europe. However, it remains unknown how V. velutina might affect native hornets when it invades Asia, where a large number of Vespa species are distributed. Thus, by analyzing the mitochondrial cytochrome c oxidase subunit I gene, we investigated whether interspecific mating occurs between V. velutina and Japanese native Vespa spp. Our results showed that the alien V. velutina causes reproductive interference in Japanese native hornet Vespa simillima. Forty-three percent of native V. simillima queens had the sperm of V. velutina males, and among the all V. simillima queens analyzed, 28% only had V. velutina sperm. We did not find evidence of V. velutina queens having the sperm of V. simillima males. These findings suggest that reproductive interference by V. velutina males poses a threat to the native V. simillima populations. A decline of V. simillima may also negatively affect other insects that interact with V. simillima.

Complete mitochondrial genome sequence of the broad-winged damselfly, Mnais pruinosa Selys, 1853 (Odonata: Calopterygidae).

In this study, we analyzed the complete mitochondrial genome of the Mnais pruinosa Selys, 1853 from Saga Prefecture, Japan. The mitochondrial genome of M. pruinosa was identified as a circular molecule of 15,494 bp, and was found to be similar to that of other damselfly species. It was predicted to contain 13 protein-coding (PCG), 22 tRNA, and two rRNA genes, as well as one A + T-rich control region. The genes ATP8 and ATP6 shared seven nucleotides, ATP6 and COIII shared one nucleotide, ND4 and ND4L shared seven nucleotides, and ND6 and Cytb shared one nucleotide. The initiation codon ATG was found in eight genes, ATC in four, and ATT in one; the termination codons TAA, TAG, incomplete TA, and single T were observed in seven, one, two, and three genes, respectively. All the tRNA genes possessed a cloverleaf secondary structure, except for tRNA-His that lacks the TΨC loop. The average AT content of mitochondrial genome was 66.18%.

A comparison of complete mitochondrial DNA sequences of Mnais costalis Selys, 1869 (Odonata: Calopterygidae) from three different populations (one allopatric and two sympatric).

In Japan, two closely-related damselflies, Mnais costalis Selys, 1869 (Odonata: Calopterygidae) and M. pruinosa Selys-Longchamps (Odonata: Calopterygidae), 1853, coexist, and they exhibit geographic variations in wing color, body size, and habitat preference. In this study, we analyzed the complete mitochondrial genome of M. costalis from Saga Prefecture, Japan (sympatric populations that exhibit wing color polymorphism), and compared the genome with M. costalis that exhibit monomorphic orange wing color. The mitochondrial genome of M. costalis from Saga Prefecture was identified as a circular molecule of 15,488 bp, similar to that found in other M. costalis populations. It was predicted to contain 13 protein-coding (PCG), 22 tRNA, and two rRNA genes, along with one A + T-rich control region. Among the PCGs, ATP8 and ATP6, ATP6 and COIII, ND4 and ND4L, and ND6 and Cytb shared seven, one, seven, and one nucleotides, respectively. The initiation codon ATG was found in eight genes, ATC in four, and ATT in one, while the termination codons TAA, TAG, TA, and T were observed in seven, one, two, and three genes, respectively. All the tRNA genes possessed a cloverleaf secondary structure, except for tRNA-His that lacks the TΨC loop. The average AT content of mitochondrial genome was 66.06%. From a phylogenetic analysis, the loss of wing color polymorphism in monomorphic sympatric populations is likely to occur with the coexistence of two Mnais species.

The complete mitochondrial genome of the yellow coloured honeybee Apis mellifera (Insecta: Hymenoptera: Apidae) of New Zealand.

The complete mitochondrial genome of the yellow coloured honeybee Apis mellifera from North Island, New Zealand was analyzed using next-generation sequencing. The mitochondrial genome was a 16,349bp circular molecule and was predicted to contain 13 protein-coding genes (PCGs), 22 tRNA genes and two rRNA genes. The initiation codon ATA was found in two genes, ATG in four genes, ATT in six genes, and ATC in one gene, while the termination codon TAA was observed in all the PCGs. Phylogenetic analysis using the sequence of 23 closely related taxa suggested a sister relationship with the Italian strain A. mellifera ligustica.

Complete mitochondrial DNA sequence of the alien hornet Vespa velutina (Insecta: Hymenoptera) invading Kyushu Island, Japan.

We analyzed the complete mitochondrial genome of the invasive Asian hornet Vespa velutina from Kyushu Island, Japan. The mitochondrial genome of V. velutina was identified as a circular molecule of 16,388 bp. We predicted that the genome contains 13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes, along with one A + T-rich control region. The average AT content is 81.68%. Molecular phylogenetic analysis using the 13 mitochondrial PCGs from 11 closely related taxa of Vespidae indicated that the V. velutina invading the Japanese Islands of Kyushu and Tsushima have a common origin.

Analysis of the complete mitochondrial genomes of two dwarf honeybee species, Apis florea and Apis andreniformis (Insecta: Hymenoptera: Apidae), in Thailand.

The dwarf honeybees Apis florea and Apis andreniformis inhabit the bush and forests of continental Asia and north Africa and some islands of Sundaland and the Philippines. We analysed, for the first time, the complete mitochondrial genomes of two dwarf honeybee species from Thailand using next-generation sequencing. Each mitochondrial genome was a circular and approximately 17 kbp molecule that included 13 protein-coding genes (PCGs), 22 tRNA genes, and two ribosomal RNA genes, along with one A + T-rich control region, besides three tRNA-Ser (AGN) repeats. The AT content values of the mitochondrial genomes of A. florea and A. andreniformis were 86.28% and 85.73%, respectively. The 1150 mutation sites in 13 PCGs differing between A. florea and A. andreniformis in Thailand were evenly distributed throughout their mitochondrial genomes. The phylogenetic relationship, inferred using 13 PCGs, was consistent with that reported in previous studies, which predicted a sister relationship between A. florea and A. andreniformis.

The complete mitochondrial genome of the bumblebee, Bombus hypocrita sapporensis (Insecta: Hymenoptera: Apidae) from Hokkaido Island, Japan.

The complete is mitochondrial genome of the Japanese bumblebee Bombus hypocrita sapporensis from Hokkaido Island, Japan is analysed using next generation sequencing. The mitochondrial genome of B. h. sapporensis was observed to be a circular molecule of 15,835 bp. The average AT content in the B. h. sapporensis mitochondrial genome was 85.53%. It was predicted to contain 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes, along with one A + T-rich control region. The PCGs had ATA, ATG, or ATT as the initiation codon and were terminated by the typical stop codon TAA, except for Cytb. All the tRNA genes typically formed a cloverleaf secondary structure, except for trnE, trnF, and trnS1. The molecular phylogenetic analysis indicated that the B. h. sapporensis from Hokkaido Island population was most similar to that of the geographically isolated B. h. sapporensis from Rebun Island.

The complete mitochondrial genome of the dusky brown-gray-colored honeybee, Apis mellifera (insecta: Hymenoptera: Apidae) of New Zealand.

We analyzed the complete mitochondrial genome of the dusky brown-gray-colored honeybee Apis mellifera, collected from North Island, New Zealand. We determined that the mitochondrial genome was a 16,336 bp and predicted 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes. The start codon ATA was found in two genes, ATG in four genes, ATT in six genes, and ATC in one gene, whereas the termination codon TAA was observed in all PCGs. The non-coding regions of tRNA-Leu and COII were consistent with the C haplotype of A. mellifera carnica. Phylogenetic analysis suggests a close relationship with the European A. mellifera.