Comparison of Gut Bacterial Communities of Locusta migratoria manilensis (Meyen) Reared on Different Food Plants.

Locusts, in particular Locusta migratoria manilensis (Meyen), have been associated with major damages in agriculture, forestry, and animal husbandry in China. At present, L. migratoria manilensis has been largely domesticated, being considered an edible insect in China. Feeding variety is one of the main characteristics of L. migratoria manilensis. It has been demonstrated that microorganisms inhabiting the insect gut impact nutrition, development, defense, and reproduction of the insect host. The aim of the present study was to search for the adaptation mechanism of L. migratoria manilensis feeding on four different food plants (goosegrass, maize leaves, soybean leaves, and pakchoi) and explore changes in the gut bacterial community structure of the insect at the fifth instar nymph stage. Proteobacteria and Firmicutes were the dominant phyla, whereas Kluyvera, Enterobacter, Pseudocitrobacter, Klebsiella, Cronobacter, Citrobacter, Lactococcus, and Weissella were the dominant genera in the gut of L. migratoria manilensis. Principal component analysis and permutational multivariate analysis of variance (PERMANOVA) revealed significant differences in the gut microbiota structure of L. migratoria manilensis fed on different food plants. Moreover, functional prediction analysis revealed that metabolic and cellular processes were the most enriched categories. Within the category of metabolic processes, the most enriched pathways were carbohydrate transport and metabolism; amino acid transport and metabolism; translation, ribosomal structure, and biogenesis; cell wall/membrane/envelope biogenesis; inorganic ion transport and metabolism; and energy production and conversion. Collectively, the present results revealed that the structure of gut bacterial communities in L. migratoria manilensis fed on different food plants is impacted by food plants, which may play an essential part in the adaptation of the host.

Cellular immune responses of the yellow peach moth, Conogethes punctiferalis (Lepidoptera: Crambidae), to the entomopathogenic fungus, Beauveria bassiana (Hypocreales: Cordycipitaceae).

The yellow peach moth (YPM), Conogethes punctiferalis, is a destructive insect pest of maize in eastern China and adapts to diverse environments, especially against pathogens. In insects, innate immunity comprising both humoral and cellular defense responses, is the primary defense against invading microbial pathogens. In this study, we identified five types of circulating hemocytes from the hemolymph of YPM larvae and analyzed their alterations and functions in immune responses to the infection of Beauveria bassiana, an entomopathogenic fungus infesting many lepidopteran species. The identified hemocytes included prohemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids. Significant decreases of total and differential hemocyte counts were recorded over time in larvae, after they were injected with B. bassiana conidia. Additionally, hemocyte-mediated phagocytosis and nodulation were initiated in the hemolymph of larvae from the B. bassiana conidia challenge. The introduction of DEAE-Sepharose Fast Flow beads stained with Congo red also induced a strong encapsulation response in the larval hemolymph. Our observations unravel the occurrence of phagocytosis, nodulation and encapsulation in the hemocoel of YPM larvae to fight against the fungal infection, and offer the first insight into the YPM immune system.

Termite colonies from mid-Cretaceous Myanmar demonstrate their early eusocial lifestyle in damp wood.

Insect eusociality is characterized by cooperative brood care, reproductive division of labour and multiple generations of adults within a colony. The morphological specializations of the different termite castes from Burmese amber were recently reported, indicating the termites possessed advanced sociality in the mid-Cretaceous. Unfortunately, all the reported Cretaceous termites are individually preserved, which does not cover the behaviours of the cooperative brood care and multiple generations of adults in the nests of the Cretaceous termites. Herein, we report three eusocial aggregations from colonies of the oldest known Stolotermitidae, Cosmotermes gen. nov., in 100 Ma mid-Cretaceous Burmese amber. One large aggregation, comprising 8 soldiers, 56 workers/pseudergates and 25 immatures of different instars, additionally presents the behaviours of cooperative brood care and overlapping generations. Furthermore, taphonomic evidence indicates Cosmotermes most probably dwelled in damp/rotting wood, which provides a broader horizon of the early societies and ecology of the eusocial Cosmotermes.

New insects feeding on dinosaur feathers in mid-Cretaceous amber.

Due to a lack of Mesozoic fossil records, the origins and early evolution of feather-feeding behaviors by insects are obscure. Here, we report ten nymph specimens of a new lineage of insect, Mesophthirus engeli gen et. sp. nov. within Mesophthiridae fam. nov. from the mid-Cretaceous (ca. 100 Mya) Myanmar (Burmese) amber. This new insect clade shows a series of ectoparasitic morphological characters such as tiny wingless body, head with strong chewing mouthparts, robust and short antennae having long setae, legs with only one single tarsal claw associated with two additional long setae, etc. Most significantly, these insects are preserved with partially damaged dinosaur feathers, the damage of which was probably made by these insects' integument-feeding behaviors. This finding demonstrates that feather-feeding behaviors of insects originated at least in mid-Cretaceous, accompanying the radiation of feathered dinosaurs including early birds.

Cretaceous winged stick insects clarify the early evolution of Phasmatodea.

Wingless and shorter winged stick insects are very common today, but most known extinct stick insects had fully developed wings, leading to contentious affinities among the extinct winged and extant groups. We report herein three male winged stick insects, assigned to Pterophasmatidae fam. nov., from mid-Cretaceous Myanmar (Burmese) amber. Pterophasmatidae fam. nov. are regarded as transitional taxa from extinct winged to modern wingless and shorter winged stick insects based on their similar tegmina venation with extinct Susumanioidea and some body features the same as extant Phasmatodea. However, their symmetric phallic organs comprising two consistent phallomeres are different from those of all living groups. Phylogenetic analyses suggest that the extinct winged taxa, including the new family, are the stem groups of modern stick and leaf insects, and all of them constitute the clade of Phasmatodea. New findings indicate winged and wingless stick insects' morphologies diversified significantly during or before the mid-Cretaceous.

Stick insect in Burmese amber reveals an early evolution of lateral lamellae in the Mesozoic.

Extant stick and leaf insects commonly imitate twigs or leaves, with lateral lamellae used to enhance crypsis or achieve mimicry for protection. However, the origin and early evolution of such lateral expansions among Phasmatodea are unknown, because all known Mesozoic phasmatodeans hitherto lack preserved evidence of such structures. We report here the first Mesozoic stick insect, Elasmophasma stictum gen. et sp. nov., with well-preserved, thin, lateral lamellae on the thoracic pleura, the terga of abdominal segments I-X and the ventrolateral margins of all femora. This new species, from the mid-Cretaceous amber of northern Myanmar, has a clear, stick-like body and is assigned to Euphasmatodea. The abdominal structures of E. stictum exhibit traces of multiple expansions of the terga, suggesting that such structure might have been an early development of body expansions used to improve crypsis for stick or leaf insects when they sprawled on twigs or leaves.

Four new species of the genus Otacilia Thorell, 1897 from Hunan Province, China (Araneae, Phrurolithidae).

Four new species of the genus Otacilia Thorell, 1897 are reported from Hunan Province, China: Otacilia hippocampasp. n., Otacilia yangmingensissp. n., Otacilia curvatasp. n., and Otacilia submicrostomasp. n. All new species are described based on both sexes. In addition, the 55 known Otacilia species are divided into four species groups.

The complete mitochondrial genome of Humphaplotropis culaishanensis sp. nov. (Orthoptera: Acridoidea: Pamphagidae: Pamphaginae).

The complete mitochondrial genome of Humphaplotropis culaishanensis sp. nov., which was collected from the Shandong Province of China, is reported here. It was 15,659 bp in length and contained 72.3% AT. All Humphaplotropis culaishanensis protein-coding sequences started with a typical ATN codon, excluding cox1 and nad6. The usual termination codon (TAN) and incomplete stop codons (T, TA) were found from 13 protein-coding genes. All tRNA genes could be folded into the typical cloverleaf secondary structure, excluding trnS(AGN) which forms another structure. The sizes of the large and small ribosomal RNA genes were 1322 and 852 bp, respectively. The AT content of the A+T-rich region was 81.3%.

A new species of the genus Utivarachna Kishida, 1940 (Araneae: Trachelidae) from China, with the first description of the male of U. fabaria Zhao & Peng, 2014 and a redescription of U. gui (Zhu, Song & Kim, 1998).

A new species belonging to the kinabaluensis group of the trachelid genus Utivarachna Kishida, 1940, U. lata sp. nov., is described from China. The male of U. fabaria Zhao & Peng, 2014 is described here for the first time based on specimens collected from Yunnan Province, China. The type material of U. gui (Zhu, Song & Kim, 1998) is re-examined and the species is redescribed.

The complete mitochondrial genome of Asiotmethis jubatus (Uvarov, 1926) (Orthoptera: Acridoidea: Pamphagidae).

The complete mitochondrial genome of Asiotmethis jubatus (Uvarov, 1926), which was collected from the Xinjiang Uygur Autonomous Region of China, is reported here. It is 15,669 bp in length and contains 72.3% AT. All Asiotmethis jubatus protein-coding sequences start with a typical ATN codon, excluding cox1 and nad6. The usual termination codon (TAN) and incomplete stop codons (T) were found from 13 protein-coding genes. All tRNA genes could be folded into the typical cloverleaf secondary structure, excluding trnS(AGN) which forms another structure. The sizes of the large and small ribosomal RNA genes are 1315 and 852 bp, respectively. The AT content of the A + T-rich region is 81.9%.

The complete mitochondrial genome of Filchnerella beicki Ramme, 1931 (Orthoptera: Acridoidea: Pamphagidae).

Abstract The complete mitochondrial genome of Filchnerella beicki Ramme, 1931, which was collected from the Ningxia Hui Autonomous Region of China, is reported here. It is 15,658 bp in length and contains 72.3% AT. All Filchnerella beicki protein-coding sequences start with a typical ATN codon, excluding cox1. The usual termination codon (TAN) and incomplete stop codons (T) were found from 13 protein-coding genes. All tRNA genes could be folded into the typical cloverleaf secondary structure, excluding trnS(AGN) which forms another structure. The sizes of the large and small ribosomal RNA genes are 1315 and 853 bp, respectively. The AT content of the A+T-rich region is 83.1%.

The complete mitochondrial genome of Gomphocerus tibetanus Uvarov, 1935 (Orthoptera: Acrididae: Gomphocerinae).

The complete nucleotide sequence of the mitochondrial genome (mitogenome) of Gomphocerus tibetanus Uvarov, 1935 (Orthoptera: Acrididae: Gomphocerinae) was determined. It is 15,571 bp in length and contains 74.8% A+T. All Gomphocerus tibetanus protein-coding sequences start with a typical ATN codon. The usual termination codons (TAA and TAG) were found from 13 PCGs except COI and COII which took incomplete codon T as termination codons. All tRNA genes could be folded into the typical cloverleaf secondary structure, except tRNA(Ser(AGN)) lacking of dihydrouridine (D) arm. The sizes of the large and small ribosomal RNA genes are 1313 and 822bp, respectively. The A+T content of the A+T-rich region is 82.3%. A preliminary analysis on characteristics of Gomphocerinae mitogenome was made by comparision among three Gomphocerinae mitogenomes and Locusta migratoria.

Initial analysis of the hemocyanin subunit type 1 (Hc1 gene) from Locusta migratoria manilensis.

Hemocyanins are copper-containing (Cu(+)) proteins that transport oxygen in many arthropods hemolymph. We characterized Hc1 gene from the grasshopper species Locusta migratoria manilensis. In particular, we cloned and sequenced the corresponding cDNAs and studied their expression at different developmental stages. The cDNA of Hc1 gene (GenBank accession no.:HQ213937) is 2271 bp in length and the open reading frame is 2016 bp, which encodes a 672 amino acids protein with a calculated molecular mass of 77.9 kD and the isoelectric point of 6.06. Sequence alignment analysis result showed that this gene shares 94.7% identity with Schistocerca americana EHP. In addition, analysis of quantitative RT-PCR indicated that, LmiHc1 was expressed in the embyro (24, 39, 62, 86, 144, and 193 h after hatch), nymphs (1st instar, 2nd instar, 3rd instar, 4th instar and 5th instar) and in adult. These results showed that Hc1 plays an important role in grasshopper, which may be related to an enhanced oxygen supply. Phylogenetic analysis of insecta based on Hc1 are basically consistent with the morphology.

The complete mitochondrial genome of Thrinchus schrenkii (Orthoptera: Caelifera, Acridoidea, Pamphagidae).

Complete nucleotide sequences of mitochondrial genome (mitogenome) of Thrinchus schrenkii (Orthoptera: Acridoidea: Pamphagidae) were determined. It is 15672 bp in length and contains 71.2% A+T. All T. schrenkii protein-coding sequences except for the cytochrome oxidase subunit I (COI) start with a typical ATN codon. Instead, CCG, which is a rare but possible initiation codon, is located at the initiation context of COI. The usual termination codons (TAA and TAG) were found from 12 PCGs. However, the ND5 had incomplete termination codon (T). All tRNA genes could be folded into the typical cloverleaf secondary structure, excluding tRNASer(AGN) which forms another structure according to the Steinberg-Cedergren tertiary structure. The sizes of the large and small ribosomal RNA genes are 1319 and 848 bp, respectively. The A+T content of the A+T-rich region is 78.7%, which is the lowest among the known mitogenome of Acridoidea.

[Molecular phylogeny of some species of the acridoidea based on 16S rDNA].

Based on the 16S rDNA of 8 species (belonging to 8 families of Acridoidea) from China and keyed to 8 relative species of Orthoptera from GenBank, the homologus sequences were compared. The used frequency of nucleotide was calculated and the molecular phylogenetic tree constructed by MEGA microsoft. In the 480 bp fragment of mitochondial 16S rDNA, A + T was about 70.7% and G + C only 29.3%. The sequence data revealed considerable variation in 188 nucleotide sites among the analyzed individuals from 8 different families. The variation rate of transversion was larger than or approach to transition. The results showed that the ranges of the 16S rDNA nucleotide divergence within species, among species of a genus, among genera of a family, and among the same family and suborder were 1.5%, 3.5%-3.6%, 4.8%-15.8% and 15.2%-25.6%, respectively. The members of the same genus and family were grouped together. The phylogenetic relationships of 8 families were Pyrgomorphidae-->Chrotogonidae-->Pamphagidae-->Oedipodidae-->Acrididae--> Arcypteridae-->Gomphoceridae-->Catantopidae. Pyrgomorphidae had closer relationships with Chrotogonidae as well as being the most ancestral group. Arcypteridae and Gomphoceridae were the sister groups and had closer relationships with most evolutionary Catantopidae. Tridactylidae was divided firstly as a suborder, which differs from the recent taxonomy distinctly. Haglidae (Tettigonoidea) and Gryllidae were grouped into suborder Ensifera, Tetrigidae and 8 families of Acridoidea were grouped into suborder Caelifera, as same as recent taxonomy.