The most common ladybird beetle, Coccinella septempunctata L., is an excellent predator of crop pests such as aphids and white flies, and it shows a wide range of adaptability, a large appetite and a high reproductive ability. Diapause research plays an important role in the artificial propagation and shelf-life extension of insect products. Although this lady beetle’s regulatory, physiological and biochemical characteristics in the diapause period are well understood, the molecular mechanism of diapause remains unknown. Therefore, we collected female adults in three different states, i.e., non-diapause, diapause and diapause termination, for transcriptome sequencing. After transcriptome sequencing using the Illumina HiSeq 2500 platform with pretreatment, a total of 417.6 million clean reads from nine samples were filtered using the program FASTX (version 0.0). Additionally, 106,262 contigs were assembled into 82,820 unigenes with an average length of 921 bp and an N50 of 1,241 bp. All of the unigenes were annotated through BLASTX alignment against the Nr or UniProt database, and 37,872 unigenes were matched. We performed further analysis of these unigenes using the Clusters of Orthologous Groups of proteins (COG), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Through pairwise comparisons of the non-diapause (ND), diapause (D), and diapause-terminated (DT) groups, 3,501 and 1,427 differentially expressed genes (DEGs) were identified between D and ND and between DT and D, respectively. Moreover, 443 of the DEGs were specifically expressed during the diapause period (i.e., DEGs that were expressed at the highest or lowest levels during diapause compared with the other stages). GO function and KEGG pathway enrichment were performed on all DEGs and showed that RNA-directed DNA polymerase activity and fatty acid metabolism were significantly affected. Furthermore, eight specific expressed genes were selected for validation using qRT-PCR. Among these eight genes, seven genes were up-regulated, and one gene was down-regulated; the change trends of the eight genes were the same between the qRT-PCR and RNA-seq analysis results. In this study, a new method for collecting and identifying diapause insects was described. We generated a vast quantity of transcriptome data from C. septempunctata L., providing a resource for gene function research. The diapause-associated genes that we identified establish a foundation for future studies on the molecular mechanisms of diapause.

Species identification of Aphytis on the basis of adult morphology is extremely difficult, especially in the A. lingnanensis group, with several cryptic species. Pupal pigmentation could be used as one of the taxonomic characters for Aphytis species, and in some instances, pupal pigmentation actually provided the first clue to the distinctness of cryptic Aphytis species. The present study investigated the full-grown larvae or younger pupae of Aphytis species, and pupal pigmentation and pupal development were observed and photographed. Four characteristic pigmentation patterns of Aphytis pupae were summarized including: entirely yellow, partly dark brown, entirely or predominantly black, and partly black. The species in the chilensis and mytilaspidis groups, and some unassigned species, generally have entirely, or predominantly and or partly black pupae. The species in the chrysomphali, funicularis, and proclia groups generally have the pupae entirely yellow. The species of the lingnanensis group have the pupae both entirely yellow, e.g., A. fisheri, and partly dark brown pupae, e.g., A. lingnanensis, A. holoxanthus and A. melinus. The pupae of Aphytis species in this study had a developmental duration of about 5–8 days at 27 ± 1 °C, 70 ± 5% RH and a photoperiod of 10L: 14D. It was found that the pupal skin was always melanized at the beginning stage, generally in the first day, among the pigmented pupae of Aphytis species. As development continued, the pigmentation became darker and the eye colour changed from pale red/brown to green. No significant intra-specific variation in pupal colour pattern was detected despite relatively high numbers of specimens examined, many from multiple, and different, origins. Overall, our study indicates that pupal pigmentation could be more helpful in species identification of Aphytis.

Two new species of Tenuipalpidae (Acari) are described from Hainan province, China: Tenuipalpus gneti sp. nov. and Tenuipalpus maai sp. nov.. The ontogenetic changes in the idiosoma and leg chaetotaxy on adults (female and male) and immature stages of T. gneti sp. nov. are presented. An old species, Tenuipalpus lineosetus Wang, 1983 is re-described with detailed illustrations based on the holotype and paratypes. A key to species of Tenuipalpus of China is provided.

Bombyx mori nucleopolyhedrovirus (BmNPV) p26 is conserved among all Lepidoptera baculoviruses that have been completely sequenced thus far, and some baculoviruses even have two copies of p26, which suggested that p26 may play an important role in the virus infection cycle. This study aimed to characterize BmNPV p26. We found that BmNPV p26 transcripts were detectable as early as 3 h post-infection (hpi), and the transcript levels rapidly increased starting from 12 hpi. Western blot analysis using an anti-p26 polyclonal antibody demonstrated that the corresponding protein was also detectable from 6 hpi in BmNPV-infected cell lysates. Immunofluorescence analysis demonstrated that p26 was mainly dispersed in the infected cell cytoplasm, whereas the over-expressed fusion protein EGFP-p26 also accumulated in the nucleus. These results indicated that p26 is an early BmNPV gene and has functions both in the cytoplasm and the nucleus. RNAi-based knockdown of p26 could produce infectious virus and normal-appearing virions but decreased budded virus (BV) production in BmNPV-infected cells at 72 hpi. Moreover, the results of further quantitative PCR (Q-PCR) analysis indicated that the gp64 and p74 transcripts levels decreased significantly. These results indicated that BmNPV p26 may be associated with BmNPV replication during the late infection stage.

Seventeen species of the mali-group of Aphelinus (Hymenoptera: Aphelinidae) are reviewed worldwide, including three new species, and five previously described species in China. The mali-group was defined by a combination of characters: head and body dark with parts of metasoma pale; fore wing with a complete row of setae inside the linea calva, or also only with a few setae in the angle between the linea and marginal vein; legs with mesocoxae, metacoxae and metatibia dark, metafemur pale. In this paper, three new species, Aphelinus tuberocephalus Wang & Huang, sp.n., Aphelinus hainanensis Wang & Huang, sp.n. and Aphelinus ruellia Wang & Huang, sp.n., are described and illustrated, with notes on one species, Aphelinus coreae Hopper & Woolley, new to China. A revised key to species of the mali-group is provided. Mitochondrial (COI) partial sequences were obtained successfully for A. tuberocephalus, A. hainanensis, A. ruellia and A. coreae.

Three new species of Baeoentedon Girault (Hymenoptera: Eulophidae) are described from China, Baeoentedon balios Wang, Huang & Polaszek sp. nov., Baeoentedon bouceki Wang, Huang & Polaszek sp. nov. and Baeoentedon virgatus Wang, Huang & Polaszek sp. nov. Both female and male of Baeoentedon balios were reared from the whitefly Pealius spina (Singh) (Hemiptera: Aleyrodidae) on the bodhi tree Ficus religiosa L. (Urticales: Moraceae). The male and the whitefly host association of Baeoentedon are recorded for the first time. A key is also provided to females of the world species of the genus.