A Chromosome-level Genome Assembly and Evolution Analysis of Andrena camellia (Hymenoptera: Andrenidae).

Andrena camellia, an effective pollinator of the economicallysignificant crop Camellia oleifera, can withstand the toxic pollen of C. oleifera, making A. camellia a crucial for resource conservation and cultivation of C. oleifera. In this study, the whole genome of A. camellia was sequenced on the Oxford Nanopore platform. The assembled genome size was 340.73 Mb including 50 scaffolds (N50=47.435 Mb) and 131 contigs (N50=17.2 Mb). A total of 11, 258 protein-coding genes were annotated, in addition, 1,104 non-coding RNAs were identified. Further analysis that some chromosomes of A. camellia have a high level of synteny with those of Apis mellifera, Osmia bicornis and Andrena minutula. Thus, our reported genome of A. camellia serves as a valuable resource for studying species evolution, behavioral biology, and adaption to toxic pollen of C. oleifera.

Lipidomic Profiling Reveals Distinct Differences in Sphingolipids Metabolic Pathway between Healthy Apis cerana cerana larvae and Chinese Sacbrood Disease.

Chinese sacbrood disease (CSD), which is caused by Chinese sacbrood virus (CSBV), is a major viral disease in Apis cerana cerana larvae. Analysis of lipid composition is critical to the study of CSBV replication. The host lipidome profiling during CSBV infection has not been conducted. This paper identified the lipidome of the CSBV-larvae interaction through high-resolution mass spectrometry. A total of 2164 lipids were detected and divided into 20 categories. Comparison of lipidome between healthy and CSBV infected-larvae showed that 266 lipid species were altered by CSBV infection. Furthermore, qRT-PCR showed that various sphingolipid enzymes and the contents of sphingolipids in the larvae were increased, indicating that sphingolipids may be important for CSBV infection. Importantly, Cer (d14:1 + hO/21:0 + O), DG (41:0e), PE (18:0e/18:3), SM (d20:0/19:1), SM (d37:1), TG (16:0/18:1/18:3), TG (18:1/20:4/21:0) and TG (43:7) were significantly altered in both CSBV_24 h vs. CK_24 h and CSBV_48 h vs. CK_48 h. Moreover, TG (39:6), which was increased by more than 10-fold, could be used as a biomarker for the early detection of CSD. This study provides evidence that global lipidome homeostasis in A. c. cerana larvae is remodeled after CSBV infection. Detailed studies in the future may improve the understanding of the relationship between the sphingolipid pathway and CSBV replication.

The complete mitogenome of Halictus (Seladonia) aerarius (Hymenoptera: Halictidae) and phylogenetic analysis.

The Halictidae, where the Seladonia aeraria belongs, was an important model organism for studying the evolution of insect social behavior. We first sequenced the complete mitochondrial genome (mitogenome) of Seladonia aeraria. The mitogenome was 15, 410 bp in length, including 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes, and a hypothetical control region. In order to reveal the phylogenetic position of Seladonia aeraria form mitogenomic level, we performed phylogenetic analysis of 13 PCGs from 15 species. The results revealed that the genus-level relationship of Halictidae was Seladonia + Lasioglossum and Seladonia aeraria was more closely related to Seladonia tumulorum than Lasioglossum sp. SJW_2017. The complete mitogenome of S. aeraria will provide a basis for further study evolution and phylogenetic analysis of Halictidae.

Comparative Mitogenomic Analysis of Two Cuckoo Bees (Apoidea: Anthophila: Megachilidae) with Phylogenetic Implications.

Bees (Hymenoptera, Apoidea and Anthophila) are distributed worldwide and considered the primary pollinators of angiosperm. Megachilidae is one of the largest families of Anthophila. In this study, two complete mitogenomes of cuckoo bees in Megachilidae, namely Coelioxys fenestrata and Euaspis polynesia, were amplified and sequenced, with a length of 17,004 bp (C. fenestrata) and 17,682 bp (E. polynesia). The obtained results show that 37 mitogenomic genes and one putative control region were conserved within Hymenoptera. Truncated stop codon T was found in the cox3 gene of E. polynesia. The secondary structure of small (rrnS) and large (rrnL) rRNA subunits contained three domains (28 helices) and five domains (44 helices) conserved within Hymenoptera, respectively. Compared with ancestral gene order, gene rearrangement events included local inversion and gene shuffling. In order to reveal the phylogenetic position of cuckoo bees, we performed phylogenetic analysis. The results supported that all families of Anthophila were monophyletic, the tribe-level relationship of Megachilidae was Osmiini + (Anthidiini + Megachilini) and Coelioxys fenestrata was clustered to the Megachile genus, which was more closely related to Megachile sculpturalis and Megachile strupigera than Euaspis polynesia.

Comparison of two criteria on the essential number calculation of Andrena camellia.

Andrena camellia Wu is one of the primary pollinators of Camellia oleifera A. in China. In this paper, the essential number of individuals for efficient pollination by this species was calculated via two criteria, based on various indicators including counts of pollen grains in provisions, from single visits, and from single foraging trips overall; single flower visit duration; single flight period duration; number of eggs laid by a single female over their lifetime; and the average number of flowers per plant. Based on the number of pollen grains collected per flower visit, the essential number of females necessary is 2107 in a 1-ha Camellia oleifera garden with 1800 plants, while only 1998 female individuals are essentially needed when estimated based on the mean number of pollen grains collected in a single flight period. We argue that the essential number estimated by the former method is more reasonable and accurate for practical applications.

The complete mitogenome of Lasioglossum affine (Hymenoptera: Halictidae) and phylogenetic analysis.

The complete mitogenome of Lasioglossum affine (Hymenoptera: Halictidae) was sequenced and analyzed. The whole mitogenome is 17,352 bp (AT%=84.1%) and encodes 37 typical eukaryotic mitochondrial genes, including 13 protein-coding genes (PCGs), 22 tRNAs, two rRNAs, and an AT-rich region. Further analysis found three gene rearrangements, where trn I-Q-M â†’ trn M-I-Q, trn W-C-Y â†’ trn C-W-Y, and trn K-D â†’ trn D-K were shuffled. The phylogenetic relationships of 19 species of Hymenoptera were established using maximum-likelihood method based on 13 concatenated PCGs. The result showed that Lasioglossum affine is a sister of Lasioglossum sp. SJW-2017.

The complete mitogenome of Nomia chalybeata (Hymenoptera: Halictidae) and phylogenetic analysis.

The complete mitogenome of Nomia chalybeata was sequenced. The mitochondrial length of N. chalybeata was 16,692 bp (AT content 85.4%), with 37 classic invertebrate mitochondrial genes (including 13 protein-coding genes, 22 transporter RNAs, and two ribosomal RNAs) and AT-rich region (AT content 91.7%). The maximum-likelihood phylogenetic relationship was constructed using 11 species from Hymenoptera. Through the phylogenetic relationship, our research team successfully used the molecular data of the mitochondrial genome to verify that N. chalybeata belongs to the family Halictidae, and also provides molecular data for the database of the family Halictidae.

The complete mitogenome of Habropoda rodoszkowskii (Hymenoptera: Apidae) and phylogenetic analysis.

The mitogenome of Habropoda rodoszkowskii, the first complete mitogenome sequence of the genus Habropoda (hymenoptera: Apidae), was sequenced. The mitogenome is 18,497 bp (The proportion of A + T in 80.7%) long, with 37 classic eukaryotic mitochondrial genes (including 13 PCGs, 22 tRNAs, and 2 rRNAs) and an AT-rich region (The proportion of A + T in 78.2%). The Bayesian-inference and Maximum-likelihood phylogenetic relationship was constructed using 15 species from Hymenoptera. According to the phylogenetic tree, Habropoda rodoszkowskii converges with genus Nomada bees (Nomada flava and Nomada flavoguttata) to be supported. In addition, Habropoda rodoszkowskii is more closely related to Apidae than to Megahilidae and Colletidae.

Characterization of mitochondrial genomes of three Andrena bees (Apoidea: Andrenidae) and insights into the phylogenetics.

Andrena is a large bee genus of >1500 species, which includes many important pollinators of agricultural systems. In this study, we present three mitochondrial genomes (mitogenomes) of Andrena species, which are the pollinators of Camellia oleifera. Compared with putative ancestral gene arrangement of insects, the three mitogenomes present identical gene rearrangement events, including local inversion (trnR) and gene shuffling (trnQ/trnM, trnK/trnD, and trnW/trnC-trnY). Most PCGs initiate with standard ATN codon and share the stop codon of TAA or TAG, whereas truncated stop codon T was detected in the atp6 gene of A. chekiangensis. Furthermore, the nad4 gene end with a single T in all three Andrena species. All tRNAs could be folded into clover-leaf secondary structure except for trnS1, with the dihydrouracil (DHU) arm forming a simple loop. Phylogenetic analysis is performed on 17 Andrena mitogenomes. Maximum likelihood and Bayesian methods generate identical topology, in which A. hunanensis and A. striata form a group and are close to A. camellia. Although A. chekiangensis is also difficult to be distinguished from A. camellia by morphological methods, A. chekiangensis and A. haemorrhoa form a clade and are grouped with the other taxa of the genus Andrena.

Phylogenetic analysis of the mitochondrial genomes in bees (Hymenoptera: Apoidea: Anthophila).

In this study, the first complete mitogenome of Andrenidae, namely Andrena camellia, is newly sequenced. It includes 13 protein-coding (PCG) genes, 22 transfer RNA (rRNA) genes, two ribosomal RNA (tRNA) genes, and a control region. Among PCGs, high conservation is observed in cytochrome oxidase genes with cox1 exhibits the highest conservation. Conversely, NADH dehydrogenase and ATPase subunit genes are more variable with atp8 presents the maximal variation. Comparison of the gene order indicates complex rearrangement in bees. Most of the rearranged events are located in the tRNA clusters of trnI-trnQ-trnM, trnW-trnC-trnY, and trnA-trnR-trnN-trnS1-trnE-trnF. Furthermore, we present the most comprehensive mitochondrial phylogeny of bee families. The monophyly of each family and the long-tongued bees is highly supported. However, short-tongued bees are inferred as paraphyletic relative to the sister relationship between Melittidae and other bee families. Furthermore, to improve the resolution of phylogeny, various datasets and analytical approaches are performed. It is indicated that datasets including third codons of PCGs facilitate to produce identical topology and higher nodal support. The tRNA genes that have typical cloverleaf secondary structures also exhibit similar positive effects. However, rRNAs present poor sequence alignment and distinct substitution saturation, which result in negative effects on both tree topology and nodal support. In addition, Gblocks treatment can increase the congruence of topologies, but has opposite effects on nodal support between the two inference methods of maximum likelihood and Bayesian inference.

Sequencing and characterization of the Megachile sculpturalis (Hymenoptera: Megachilidae) mitochondrial genome.

The mitochondrial genome (mitogenome) of Megachile sculpturalis (Hymenoptera: Megachilidae) has been reported in this study. This is the first sequenced mitogenome from the family Megachilidae. The sequenced region of this mitochondrial genome is determined to be 16 581 bp in length, including 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a partial sequence of the AT-rich region, and the AT-rich region contains several characteristic repeated sequences. Additionally, we conducted a phylogenetic analysis using all the mitochondrial genomes of Apoidea that are available in Genbank. Bayesian method supported the monophyly of both Apidae and Colletidae. And Megachilidae was inferred as the sister group of Apidae.

The complete mitochondrial genome of the Colletes gigas (Hymenoptera: Colletidae: Colletinae).

The complete mitochondrial genome of the Colletes gigas (Hymenoptera: Colletidae) is determined to be 15,885 bp in length, containing 37 typical animal mitochondrial genes: 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a major non-coding AT-rich region. All PCGs initiate with typical ATN codons and end with the complete termination codon TAA, except for the cob gene, which harbors the stop codon of TAG. Twenty-four intergenic spacers (614 bp in total) and 7 overlapping regions (37 bp in total) are dispersed throughout the whole genome. The non-coding AT-rich region is 539 bp long and contains a tandem repeat region, which has also been reported in other insects. This is the first completely sequenced mitochondrial genome of the family Colletidae.

The complete mitochondrial genome of Melanargia asiatica (Lepidoptera: Nymphalidae: Satyrinae).

We sequenced the complete mitochondrial genome of Melanargia asiatica (Lepidoptera: Nymphalidae: Satyrinae). The entire closed circular molecule is 15,142 bp long, containing 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a AT-rich region. All protein-coding genes (PCGs) initiate with the typical start codons ATN, with the exception of cox1, which uses CGA instead. Nine PCGs use the conventional stop codons (TAA) and the other four genes (cox1, cox2, nad4 and nad5) use a single T as the stop codon. All tRNA genes display typical secondary cloverleaf structures, except for trnS1 (AGN), whose dihydrouridine (DHU) arm is replaced by a simple loop, as observed in all other lepidopterans. The AT-rich region is 319 bp in length and contains some features characteristic of lepidopterans, such as the ATAGA motif followed by a 19-bp poly-T stretch and a microsatellite-like repeat of (TA)6T(TA) preceded by the ATTTA motif.

Sequencing and characterization of mitochondrial genome of Eurytoma sp. (Hymenoptera: Eurytomidae).

The mitochondrial genome (mitogenome) of Eurytoma sp. (Hymenoptera: Eurytomidae: Eurytominae) was determined in this study. The sequenced region is 14,946 bp with an A + T content of 82.65%, including 13 protein-coding genes, 2 ribosomal RNAs, 18 transfer RNAs, and a partial sequence of AT-rich region. All PCGs begin with typical ATN codons and use standard canonical TAA as their termination codons, except for atp6 gene, which stops with an incomplete codon T. A comparison of all the sequenced hymenopteran mitogenomes revealed that Eurytoma sp. has dramatic gene rearrangements, some of which is peculiar to Chalcidoidea. What's more, Maximum likelihood (ML) analysis highly supported the monophyly of both Chalcidoidea and Apocrita.

Sequencing and characterization of the Megachile strupigera (Hymenoptera: Megachilidae) mitochondrial genome.

The mitochondrial genome (mitogenome) of Megachile strupigera (Hymenoptera: Megachilidae: Megachilinae) was determined in our study. The sequenced region is 15,193 bp with an A + T content of 83.44%, including 13 protein-coding genes, two ribosomal RNAs and 19 transfer RNAs. All PCGs are initiated by typical ATN codons and stop with the complete termination codon TAA, except for nad3 gene, which has an incomplete stop codon T. Bayesian method supported the monophyly of both Megachilidae and Apoidea. And within the Apoidea, Apidae and Megachilidae formed a sister clade to Colletidae.

The complete mitochondrial genome of Blue Pansy, Junonia orithya (Lepidoptera: Nymphalidae: Nymphalinae).

The complete mitochondrial genome (mitogenome) of Junonia orithya Linnaeus (Lepidoptera: Nymphalidae: Nymphalinae) is determined to be 15,214 bp in length, including 37 typical mitochondrial genes and an AT-rich region. Its gene order and orientation are identical to those of other butterfly species. All PCGs are initiated by typical ATN codons, except for cox1 gene which is started by CGA codon. Nine genes use complete termination codon (TAA), whereas the cox1. cox2. nad1 and nad4 genes end with single T. Except for trnS1(AGN), all tRNA genes display typical secondary cloverleaf structures as those of other insects. The 331 bp long AT-rich region contains several features common to the other lepidopterans, such as the ATAGA motif followed by a 18 bp poly-T stretch, two microsatellite-like (TA)9 elements, a 5 bp poly-A stretch immediately upstream of trnM gene.

The first mitochondrial genome for the butterfly family Riodinidae (Abisara fylloides) and its systematic implications.

The Riodinidae is one of the lepidopteran butterfly families. This study describes the complete mitochondrial genome of the butterfly species Abisara fylloides, the first mitochondrial genome of the Riodinidae family. The results show that the entire mitochondrial genome of A. fylloides is 15 301 bp in length, and contains 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and a 423 bp A+T-rich region. The gene content, orientation and order are identical to the majority of other lepidopteran insects. Phylogenetic reconstruction was conducted using the concatenated 13 protein-coding gene (PCG) sequences of 19 available butterfly species covering all the five butterfly families (Papilionidae, Nymphalidae, Peridae, Lycaenidae and Riodinidae). Both maximum likelihood and Bayesian inference analyses highly supported the monophyly of Lycaenidae+Riodinidae, which was standing as the sister of Nymphalidae. In addition, we propose that the riodinids be categorized into the family Lycaenidae as a subfamilial taxon. The Riodinidae is one of the lepidopteran butterfly families. This study describes the complete mitochondrial genome of the butterfly species Abisara fylloides, the first mitochondrial genome of the Riodinidae family. The results show that the entire mitochondrial genome of A. fylloides is 15 301 bp in length, and contains 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and a 423 bp A+T-rich region. The gene content, orientation and order are identical to the majority of other lepidopteran insects. Phylogenetic reconstruction was conducted using the concatenated 13 protein-coding gene (PCG) sequences of 19 available butterfly species covering all the five butterfly families (Papilionidae, Nymphalidae, Peridae, Lycaenidae and Riodinidae). Both maximum likelihood and Bayesian inference analyses highly supported the monophyly of Lycaenidae+Riodinidae, which was standing as the sister of Nymphalidae. In addition, we propose that the riodinids be categorized into the family Lycaenidae as a subfamilial taxon.

The complete mitochondrial genome of Cupido argiades (Lepidoptera: Lycaenidae).

The complete mitogenome of Cupido argiades (Lepidoptera: Lycaenidae) was determined in this study. The genome is 15,330 bp long, presenting a typical gene organization and order for completely sequenced lepidopoteran mitogenomes. All protein-coding genes (PCGs) are initiated by ATN codons, except for cox1, which uses CGA as its start codon. Some PCGs harbor TAG (nad3) or incomplete termination codon T (cox1, cox2, nad5), while others use standard canonical TAA as their termination codons. Furthermore, the largest noncoding A+T-rich region with the length of 450 bp contains two microsatellite-like repeats of (TA)9 and a conserved motif ATAGA followed by a 19-bp poly-T stretch.

Camellia fruit borer, Neoblastobasis camelliae, a new species of Blastobasinae in China (Lepidoptera, Blastobasidae).

A new Neoblastobasis Kuznetzov & Sinev, 1985, Neoblastobasis camelliae Chen & Wu, sp. nov., is described from Jiangxi, China, and reported feeding on the fruits of Camellia oleifera Abel (Theaceae). Illustrations of the adult antennal base, tergal spines on the posterior part of tergites, male genital structures, and notes on its biology are provided. A list of the Blastobasidae known from China is also included. Two species, Blastobasis murcyae Adamski & Brown, 2002 and B. ianella Adamski, 2003 are transferred from Blastobasis to Neoblastobasis.

The complete mitochondrial genome of the yellow coaster, Acraea issoria (Lepidoptera: Nymphalidae: Heliconiinae: Acraeini): sequence, gene organization and a unique tRNA translocation event.

In this paper, the complete mitochondrial genome of Acraea issoria (Lepidoptera: Nymphalidae: Heliconiinae: Acraeini) is reported; a circular molecule of 15,245 bp in size. For A. issoria, genes are arranged in the same order and orientation as the complete sequenced mitochondrial genomes of the other lepidopteran species, except for the presence of an extra copy of tRNA(Ile(AUR)b) in the control region. All protein-coding genes of A. issoria mitogenome start with a typical ATN codon and terminate in the common stop codon TAA, except that COI gene uses TTG as its initial codon and terminates in a single T residue. All tRNA genes possess the typical clover leaf secondary structure except for tRNA(Ser(AGN)), which has a simple loop with the absence of the DHU stem. The sequence, organization and other features including nucleotide composition and codon usage of this mitochondrial genome were also reported and compared with those of other sequenced lepidopterans mitochondrial genomes. There are some short microsatellite-like repeat regions (e.g., (TA)(9), polyA and polyT) scattered in the control region, however, the conspicuous macro-repeats units commonly found in other insect species are absent.