Comparative Analysis of Mitogenomes of Chironomus (Diptera: Chironomidae).

(1) Background: Chironomids are biological indicators, playing an important role in monitoring and assessing the changes in water ecosystems. Mitochondrial genomes have been widely applied as a molecular marker to analyze the taxonomy and phylogeny of insects. However, knowledge of the mitogenomes of Chironomus species is scarce at present, which limits our understanding of the evolutionary relationships among Chironomus. (2) Methods: In our study, the mitogenomes and their basic structure of 12 Chironomus species and one Microchironomus species were newly sequenced. Combined with reported mitogenomes, a total of 15 mitogenomes of Chironomus were selected for a comparative mitogenomic analysis and phylogenetic reconstruction of Chironomus. (3) Results: Each mitogenome of the Chironomus species has the typical 37 genes and a control region. The basic structure of the whole mitogenomes of Chironomus species is relatively conservative, and the genetic arrangements stay the same as the ancestral mitogenome. (4) Conclusions: Our study enriches the library of mitogenomes of chironomids and provides a valuable resource for understanding the evolutionary history of Chironomus.

Antifungal Substances Produced by Xenorhabdus bovienii and Its Inhibition Mechanism against Fusarium solani.

Fungal colonization can severely damage artifacts. Nematode endosymbiotic bacteria exhibit good prospects in protecting artifacts from fungal damage. We previously found that supernatant from the fermentation of nematode endosymbiotic bacterium, Xenorhabdus bovienii, is effective in inhibiting the growth of Fusarium solani NK-NH1, the major disease fungus in the Nanhai No.1 Shipwreck. Further experiments proved that X. bovienii produces volatile organic compounds (VOCs) that inhibit NK-NH1. Here, using metabolomic analysis, GC-MS, and transcriptomic analysis, we explored the antifungal substances and VOCs produced by X. bovienii and investigated the mechanism underlying its inhibitory effect against NK-NH1. We show that X. bovienii produces several metabolites, mainly lipids and lipid-like molecules, organic acids and derivatives, and organoheterocyclic compounds. The VOCs produced by X. bovienii showed two specific absorption peaks, and based on the library ratio results, these were predicted to be of 2-pentanone, 3-(phenylmethylene) and 1-hexen-3-one, 5-methyl-1-phenyl. The inhibition of F. solani by VOCs resulted in upregulation of genes related to ribosome, ribosome biogenesis, and the oxidative phosphorylation and downregulation of many genes associated with cell cycle, meiosis, DNA replication, and autophagy. These results are significant for understanding the inhibitory mechanisms employed by nematode endosymbiotic bacteria and should serve as reference in the protection of artifacts.

Evaluation of indigenous entomopathogenic nematodes in Southwest China as potential biocontrol agents against Spodoptera litura (Lepidoptera: Noctuidae).

Spodoptera litura is a notorious leaf feeding insect pest in the Asia-Pacific region and leads to a significant economic loss in vegetable and field crop production. Entomopathogenic nematodes (EPNs), lethal parasites of insects, are used as biocontrol agents. Yunnan Province in China is a well-known region due to its rich biodiversity. In the present study, a survey of EPNs using the Galleria-baiting technique was conducted in 2017 and 2018 throughout the entire Yunnan province. In total, 789 soil samples were collected from 232 sites, of which 75 samples were positive for EPNs. Phylogenetic analyses of ITS, D2D3 expansion region of the 28S rRNA gene, as well as mitochondrial cytochrome c oxidase subunit I (COI), were performed to identify isolated nematode species and evaluate their genetic diversity. In total, 13, 3, and 58 identified populations belong to Steinernema, Heterorhabditis, and Oscheius, respectively. The phylogenetic relationships of EPN species in the three genera were analyzed with the Neighbor-Joining method. The virulence of the trapped isolates in the genera of Steinernema, Heterorhabditis, and Oscheius against S. litura was evaluated. Ten new indigenous isolates from Steinernema and Heterorhabditis showed prominent virulence to S. litura within 48 hr which is equivalent to that of commercial EPNs populations. The present study provides background information on indigenous EPN resources for S. litura control in Asia-Pacific region.

First report of Ovomermis sinensis (Nematoda: Mermithidae) parasitizing fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae) in China.

Spodoptera frugiperda invaded China in the end of 2018 and has caused severe damage to maize and other crops. Several S. frugiperda naturally parasitized by nematodes were observed in Hainan Province, China. The morphological characteristics based on the results of scanning electron microscopy indicated that the nematode belongs to the family Mermithidae. Additionally, coding sequences for the 18 S and 28 S rDNA were amplified from the nematode genome, and phylogenetic analysis revealed that the nematode belongs to Ovomermis sinensis, a known entomoparasitic nematode. Our finding is the first record that S. frugiperda was naturally parasitized by O. sinensis. The results of this study are of great significance for potential biological control of S. frugiperda by indigenous natural beneficial organisms, i.e. O. sinensis within an integrated pest management system.Spodoptera frugiperda invaded China in the end of 2018 and has caused severe damage to maize and other crops. Several S. frugiperda naturally parasitized by nematodes were observed in Hainan Province, China. The morphological characteristics based on the results of scanning electron microscopy indicated that the nematode belongs to the family Mermithidae. Additionally, coding sequences for the 18 S and 28 S rDNA were amplified from the nematode genome, and phylogenetic analysis revealed that the nematode belongs to Ovomermis sinensis, a known entomoparasitic nematode. Our finding is the first record that S. frugiperda was naturally parasitized by O. sinensis. The results of this study are of great significance for potential biological control of S. frugiperda by indigenous natural beneficial organisms, i.e. O. sinensis within an integrated pest management system.

New or little-known Diamesinae (Diptera: Chironomidae) from Oriental China.

Morphological description for adult male of a new species Diamesa qiangi sp. nov. and redescription for adult males of the little-known species Linevitshia prima Makarchenko and Sasayusurika nigatana (Tokunaga) of subfamily Diamesinae from the Oriental Region of China are given.

The symbiotic bacteria Alcaligenes faecalis of the entomopathogenic nematodes Oscheius spp. exhibit potential biocontrol of plant- and entomopathogenic fungi.

Soil-dwelling entomopathogenic nematodes (EPNs) kill arthropod hosts by injecting their symbiotic bacteria into the host hemolymph and feed on the bacteria and the tissue of the dying host for several generations cycles until the arthropod cadaver is completely depleted. The EPN-bacteria-arthropod cadaver complex represents a rich energy source for the surrounding opportunistic soil fungal biota and other competitors. We hypothesized that EPNs need to protect their food source until depletion and that the EPN symbiotic bacteria produce volatile and non-volatile exudations that deter different soil fungal groups in the soil. We isolated the symbiotic bacteria species (Alcaligenes faecalis) from the EPN Oscheius spp. and ran infectivity bioassays against entomopathogenic fungi (EPF) as well as against plant pathogenic fungi (PPF). We found that both volatile and non-volatile symbiotic bacterial exudations had negative effects on both EPF and PPF. Such deterrent function on functionally different fungal strains suggests a common mode of action of A. faecalis bacterial exudates, which has the potential to influence the structure of soil microbial communities, and could be integrated into pest management programs for increasing crop protection against fungal pathogens.

Exploring the utility of DNA barcoding in species delimitation of Polypedilum (Tripodura) non-biting midges (Diptera: Chironomidae).

In this study, we tested the utility of the mitochondrial gene cytochrome c oxidase subunit 1 (CO1) as the barcode region to deal with taxonomical problems of Polypedilum (Tripodura) non-biting midges (Diptera: Chironomidae). The 114 DNA barcodes representing 27 morphospecies are divided into 33 well separated clusters based on both Neighbor Joining and Maximum Likelihood methods. DNA barcodes revealed an 82% success rate in matching with morphospecies. The selected DNA barcode data support 37-64 operational taxonomic units (OTUs) based on the methods of Automatic Barcode Gap Discovery (ABGD) and Poisson Tree Process (PTP). Furthermore, a priori species based on consistent phenotypic variations were attested by molecular analysis, and a taxonomical misidentification of barcode sequences from GenBank was found. We could not observe a distinct barcode gap but an overlap ranged from 9-12%. Our results supported DNA barcoding as an ideal method to detect cryptic species, delimit sibling species, and associate different life stages in non-biting midges.

Two new species and one new record of Parakiefferiella Thienemann, 1936 from China (Diptera, Chironomidae).

Two new species of the genus Parakiefferiella Thienemann, 1936 (Parakiefferiella fasciata Liu & Wang, sp. n. and Parakiefferiella liupanensis Liu & Wang, sp. n.) are described as adult males from China. Parakiefferiella tamatriangulata Sasa is recorded from China for the first time. A key to the known adult males of Parakiefferiella from China is presented. The generic diagnosis of male Parakiefferiella given by Cranston et al. (1989) is emended.