Two new species of the genus Kingdonella Uvarov, 1933 (Orthoptera, Acridoidea) and the first reported male of Kingdonellaqinghaiensis Zheng, 1990.

Two new species of the genus Kingdonella and the first report of a male K.qinghaiensis Zheng, 1990 are presented. The new species K.gandensis sp. nov. has similar morphological features to K.wardi Uvarov, 1933, but it differs from the latter in having 1) the hind tibia black; 2) the epiproct, in males, with a median groove in the basal 1/2 and in the apical 1/4; 3) the denticles of the male epiproct black; 4) the outside of the hind femur reddish-brown on the basal 1/4 and black on the apical 3/4; and 5) the ventral face of the hind femur black on the outer side. The second new species, K.biruensis sp. nov., is morphologically close to K.pienbaensis Zheng, 1980 but differs from the latter in having 1) the length of the middle segment (12th segment) of antennae 1.2 times longer than its width; 2) the subgenital plate sharp-cornered in males; 3) the ovipositor smooth; 4) the upper half of hind femur outside surface with two black spots; and 5) the ventral face of the hind femur black on its outer side, red on the basal 2/3, and black on the apical 1/3 of its inner side. Finally, we provide a key to all known species of Kingdonella.

Gut microorganisms of Locusta migratoria in various life stages and its possible influence on cellulose digestibility.

Locusta migratoria is an important phytophagous pest, and its gut microbial communities play an important role in cellulose degradation. In this study, the gut microbial and cellulose digestibility dynamics of Locusta migratoria were jointly analyzed using high-throughput sequencing and anthrone colorimetry. The results showed that the gut microbial diversity and cellulose digestibility across life stages were dynamically changing. The species richness of gut bacteria was significantly higher in eggs than in larvae and imago, the species richness and cellulose digestibility of gut bacteria were significantly higher in early larvae (first and second instars) than in late larvae (third to fifth instars), and the diversity of gut bacteria and cellulose digestibility were significantly higher in imago than in late larvae. There is a correlation between the dynamics of gut bacterial communities and cellulose digestibility. Enterobacter, Lactococcus, and Pseudomonas are the most abundant genera throughout all life stages. Six strains of highly efficient cellulolytic bacteria were screened, which were dominant gut bacteria. Carboxymethyl cellulase activity (CMCA) and filter paper activity (FPA) experiments revealed that Pseudomonas had the highest cellulase enzyme activity. This study provides a new way for the screening of cellulolytic bacteria and lays the foundation for developing insects with significant biomass into cellulose-degrading bioreactors. IMPORTANCE: Cellulose is the most abundant and cheapest renewable resource in nature, but its degradation is difficult, so finding efficient cellulose degradation methods is an urgent challenge. Locusta migratoria is a large group of agricultural pests, and the large number of microorganisms that inhabit their intestinal tracts play an important role in cellulose degradation. We analyzed the dynamics of Locusta migratoria gut microbial communities and cellulose digestibility using a combination of high-throughput sequencing technology and anthrone colorimetry. The results revealed that the gut microbial diversity and cellulose digestibility were dynamically changed at different life stages. In addition, we explored the intestinal bacterial community of Locusta migratoria across life stages and its correlation with cellulose digestibility. The dominant bacterial genera at different life stages of Locusta migratoria were uncovered and their carboxymethyl cellulase activity (CMCA) and filter paper activity (FPA) were determined. This study provides a new avenue for screening cellulolytic bacteria and lays the foundation for developing insects with significant biomass into cellulose-degrading bioreactors.

Comparative analysis of mitogenomes among three species of grasshoppers (Orthoptera: Acridoidea: Gomphocerinae) and their phylogenetic implications.

Whole mitochondrial genomes have been widely used in phylogenetic analysis, population genetics and biogeography studies. This study sequenced and characterized three complete mitochondrial genomes (Dasyhippus peipingensis, Myrmeleotettix palpalis, Aeropedellus prominemarginis) and determined their phylogenetic position in Acrididae. The length of the mitochondrial genomes ranged from 15,621-15,629 bp and composed of 13 PCGs, 2 rRNA, 22 tRNA genes and an AT control region. The arrangement and structure of the mitochondrial genomes were similar to those of other invertebrates. Comparative genomics revealed that the three mitochondrial genomes were highly conserved in terms of gene size, structure, and codon usage, all PCGs were purified selections with an ATN start codon and a TAN stop codon. All tRNAs could be folded into the typical clover-leaf structure, except tRNA Ser (AGN) that lacked a dihydrouridine (DHU) arm. Phylogenetic analysis based on 13 PCGs of 34 Acrididae species and seven outgroup species revealed that differences in the shape of antennae within the family Acrididae should be given less weight as a taxonomic character for higher-level classification. Moreover, the divergence time estimates indicates that in Gomphocerinae, the species with clubbed antennae were formed within the nearest 18 Mya, and Pacris xizangensis is more ancient.

Isolation and characterization of intestinal bacteria associated with cellulose degradation in grasshoppers (Orthoptera).

Insect gut bacteria play an essential role in the nutritional metabolism, growth, and development of insects. Grasshoppers (Orthoptera) are cellulose-rich plant-feeding pests. Although the biological potential of grasshopper gut microorganisms to assist cellulose decomposition is well established, microbial resources for efficient degradation of cellulose biomass are still scarce and need to be developed. In this study, we used selective media to isolate cellulose-degrading bacteria from the intestines of Atractomorpha sinensis, Trilophidia annulata, Sphingonotus mongolicus, and Calliptamus abbreviatus. Phylogenetic analysis based on the maximum likelihood method using 16S rDNA sequencing sequences to identify bacteria revealed the isolation of 11 strains belonging to 3 genera, including Klebsiella, Aeromonas, and Bacillus. The degradability of the isolates to cellulose was then determined by the DNS colorimetric method, and the results showed that Bacillus had the highest degradation rate. The elucidation of microbial cellulose degradation capacity in grasshoppers not only contributes to the understanding of multiple plant-insect-microbe interactions, but also provides a valuable microbial resource for solving the biomass conversion of cellulose species problem.

Microbial gut diversity in four grasshopper species and its correlation with cellulose digestibility.

Grasshoppers are common pests, and their intestinal microbes have coevolved with them. These microorganisms have varied community structures, and they participate in the nutritional absorption and metabolism of grasshoppers. Here, we describe the gut microbiota diversity of four species of grasshoppers, Oxya chinensis, Pararcyptera microptera meridionalis, Gastrimargus marmoratus, and Calliptamus abbreviatus. We constructed a 16S rDNA gene library and analyzed the digestibility of cellulose and hemicellulose in grasshoppers using moss black phenol and anthrone colorimetry. The grasshopper with the highest microbial diversity in the gut among the four species was Oxya chinensis, and there were no significant differences in gut microbial diversity between the two geographic collections of Oxya chinensis. The most dominant phyla of the four grasshopper gut microorganisms were Proteobacteria, Bacteroidetes, and Firmicutes, and the most dominant genus was Enterobacter. The gut microbiota features of the four grasshoppers were correlated with their cellulose and hemicellulose digestibility. There was a significant positive correlation with cellulose digestibility for Pantoea. A significant negative correlation was found with cellulose digestibility for Acinetobacter, Enterococcus, Citrobacter, Serratia. A significant negative correlation was found with hemicellulose digestibility for Pantoea. This study contributes to the understanding of the structural composition of different species of grasshoppers gut microbiota, which may be useful for developing grasshopper digestive tracts as bioreactors for cellulose decomposition, improving the decomposition and utilization of agricultural straw, producing clean biomass energy, and processing biologically derived products.

Analysis of Intestinal Microbial Diversity of Four Species of Grasshoppers and Determination of Cellulose Digestibility.

Grasshoppers (Insecta, Orthoptera, Acridoidea) are a large group of agricultural and animal husbandry pests. They have a large food intake with high utilization of plants fibers. However, the composition of the grasshopper gut microbial community, especially the relationship between gut microbial community and cellulose digestibility, remains unclear. In this research, 16S rRNA gene sequences were used to determine the intestinal microbial diversity of Acrida cinerea, Trilophidia annulata, Atractomorpha sinensis and Sphingonotus mongolicus, and Spearman correlation analysis was performed between the intestinal microbes of grasshoppers and the digestibility of cellulose and hemicellulose. The results showed that Proteobacteria was the dominant phylum and Klebsiella was the dominant genus in the guts of the four species of grasshoppers; there was no significant difference in the species composition of the gut microbes of the four species of grasshoppers. Spearman correlation analysis showed that Brevibacterium and Stenotrophomonas were significantly correlated with cellulose digestibility. Brevibacterium, Clavibacter, Microbacterium and Stenotrophomonas were significantly associated with hemicellulose digestibility. Our results confirmed that the gut microbes of grasshoppers were correlated with the digestibility of cellulose and hemicellulose, and indicated that grasshoppers may have the potential to develop into bioreactors, which can be applied to improve straw utilization efficiency in the future.

The potential application of vegetable oils in the D-xylose and L-cysteine Maillard reaction system for meaty aroma production.

Vegetable oils with different unsaturated degrees (corn > peanut > olive > palm > coconut) and corresponding enzymatic hydrolysis-thermal oxidation (ENTH) oils were added into D-xylose and L-cysteine Maillard reaction (MR) system, respectively, for potential meaty aroma production. Results indicated that coconut oil and palm oil had lower peroxide, p-anisidine, total antioxidant values, and malondialdehyde content after ENTH and MR. Corn, peanut, and olive oils after ENTH could significantly promote MR because they caused the lower pH, higher reactants depletion, and higher browning degree in the water phase separated from the oil-Maillard reaction system. Additionally, the volatiles were mainly formed from oil phase. Corn, peanut, and olive oils were beneficial to volatiles formation in the oil-MR system due to fatty acid oxidation, especially oleic acid and linoleic acid. Meanwhile, compared to raw oils, adding ENTH oils to the MR system could reduce the content of oxygen, nitrogen, and sulfur-containing heterocycles.

Comparative Analysis of Mitogenomes among Five Species of Filchnerella (Orthoptera: Acridoidea: Pamphagidae) and Their Phylogenetic and Taxonomic Implications.

Mitogenomes have been widely used for exploring phylogenetic analysis and taxonomic diagnosis. In this study, the complete mitogenomes of five species of Filchnerella were sequenced, annotated and analyzed. Then, combined with other seven mitogenomes of Filchnerella and four of Pamphagidae, the phylogenetic relationships were reconstructed by maximum likelihood (ML) and Bayesian (BI) methods based on PCGs+rRNAs. The sizes of the five complete mitogenomes are Filchnerella sunanensis 15,656 bp, Filchnerella amplivertica 15,657 bp, Filchnerella nigritibia 15,661 bp, Filchnerella pamphagoides 15,661 bp and Filchnerella dingxiensis 15,666 bp. The nucleotide composition of mitogenomes is biased toward A+T. All tRNAs could be folded into the typical clover-leaf structure, except that tRNA Ser (AGN) lacked a dihydrouridine (DHU) arm. The phylogenetic relationships of Filchnerella species based on mitogenome data revealed a general pattern of wing evolution from long wing to increasingly shortened wing.

A new species of the genus Sulcohumpacris Yin, Yin et Cao, 2017 (Orthoptera, Acridoidea, Pamphagidae) from China.

A new species of the genus Sulcohumpacris Yin, Yin et Cao, 2017 i.e. Sulcohumpacris wuanensis sp. nov. is described in this paper from China. The new species is similar to Sulcohumpacris hebeiensis Yin, Yin et Cao, 2017 but differs from latter by median keel of pronotum cut by hind sulcus slightly, tegmen of male cover 2/3 tympanum, length of Krauss' organ 1.8 times of width in female, apex of subgenital plate of male point, cercus of male narrow at base, lower margin of Epiphallus with large projection in the middle and hind knee of female not curved at lower margin. Type specimens are deposited in the College of Life Sciences, Hebei University, Baoding, China.

Diversity of the gut microbiome in three grasshopper species using 16S rRNA and determination of cellulose digestibility.

BACKGROUND: Grasshoppers are typical phytophagous pests, and they have large appetites with high utilization of plants fibers, the digestion of which may depend on the microorganisms in their intestines. Grasshoppers have the potential to be utilized in bioreactors, which could improve straw utilization efficiency in the future. In this study, we describe the gut microbiome in three species of grasshoppers, Oedaleus decorus asiaticus, Aiolopus tamulus and Shirakiacris shirakii, by constructing a 16S rDNA gene library and analyzed the digestibility of cellulose and hemicellulose in the grasshoppers by using moss black phenol colorimetry and anthrone colorimetry. RESULTS: There were 509,436 bacterial OTUs (Operational Taxonomic Units) detected in the guts of all the grasshoppers sampled. Among them, Proteobacteria and Firmicutes were the most common, Aiolopus tamulus had the highest bacterial diversity, and Shirakiacris shirakii had the highest bacterial species richness. The intestinal microflora structure varied between the different species of grasshopper, with Aiolopus tamulus and Shirakiacris shirakii being the most similar. Meanwhile, the time at which grasshopper specimens were collected also led to changes in the intestinal microflora structure in the same species of grasshoppers. Klebsiella may form the core elements of the microflora in the grasshopper intestinal tract. The digestibility of cellulose/hemicellulose among the three species grasshoppers varied (38.01/24.99%, 43.95/17.21% and 44.12/47.62%). LEfSe analysis and Spearman correlation coefficients showed that the hemicellulosic digestibility of Shirakiacris shirakii was significantly higher than that of the other two species of grasshopper, which may be related to the presence of Pseudomonas, Stenotrophomonas, Glutamicibacter, Corynebacterium, and Brachybacterium in Shirakiacris shirakii intestinal tract. CONCLUSION: The intestinal microbial communities of the three grasshoppers species are similar on phylum level, but the dominant genera of different species grasshoppers are different. The cellulose digestibility of the three species of grasshoppers is relatively high, which may be correlated with the presence of some gut microbiome. Increasing the understanding of the structure and function of the grasshopper intestinal microflora will facilitate further research and the utilization of intestinal microorganisms in the future.

Comparative analysis of mitogenomes among three species of Haplotropidini grasshoppers and a new species of the genus Sulcotropis (Orthoptera: Acridoidea: Pamphagidae) from China.

The complete mitochondrial genomes of three species of Haplotropidini were sequenced, annotated and analyzed. Then, combined with 18 species mitogenomes of Acridoidea and 1 species of Tridactyloidea, the phylogenetic relationships were reconstructed by maximum likelihood (ML) and Bayesian (BI) methods based on PCGs. The phylogenetic relationship tree showing that Sulcotropis Yin et Chou is a valid genus and not a synonym of Haplotropis Saussure, 1888. A new species Sulcotropis xiaowutaiensis sp. nov. is described in this paper from China, it is allied to Sulcotropis cyanipes Yin et Chou, 1979, but differs from latter by median carina of pronotum cut by posterior transverse sulcus slightly, epiphallus with middle part equal both sides in high, cercus of male gradually widened at base, interspace of mesosternum narrowed in the base slightly and subgenital plate of female oblong, hind margin with small acute angle in the middle. Type specimens are deposited in the College of Life Sciences, Hebei University, Baoding, Hebei, China.

Comparative analysis of mitogenomes among six species of grasshoppers (Orthoptera: Acridoidea: Catantopidae) and their phylogenetic implications in wing-type evolution.

The degree of wing development has a close relationship with insects' movement ability and range, and it should also be closely related to mitochondrial-related genes. The complete mitochondrial genomes of six species of Catantopidae were sequenced, annotated and analyzed. Then, combined with 37 mitogenomes of grasshoppers, the ratio of nonsynonymous substitution to synonymous substitution (Ka/Ks) of the combined sequences of protein coding genes (PCGs) was calculated by DnaSP5, and the phylogenetic relationships were reconstructed by maximum likelihood (ML) and Bayesian (BI) methods based on PCGs+rRNAs. The results showed that the sizes of the six complete mitogenomes are Stenocatantops mistshenkoi Willemse F., 1968, 15,573 bp; Traulia lofaoshana Tinkham, 1940, 15,645 bp; Sinopodisma rostellocerca You, 1980, 15,622 bp; Anapodisma miramae Dovnar-Zapolskij, 1932, 15,189 bp; Qinlingacris elaeodes Yin & Chou, 1979, 15,221 bp; and Eozubovskya planicaudata Zhang & Jin, 1985, 15,830 bp; their structures are the same as those of Acridoidea. The AT bias of the wing-degenerated group (lobiform and apterous) is higher than that of the longipennate group, and more nonsynonymous substitutions accumulated in the wing-degenerated group than in the longipennate group (P = 0.000), which indicates that the wing-degenerated group has undergone weaker evolutionary selection than the longipinnate group. The phylogenetic tree shows that the wing-degenerated group in the Catantopidae are multiorigin and present parallel evolution.

A new species of the genus Criotettix Bolivar (Orthoptera:Tetrigoidea:Scelimenidae) from Zhejiang, China.

A new species of the genus is described from Zhejiang, China in this paper. The new species Criotettix zhejiangensis sp. nov. is similar to Criotettix strivertexoides Zheng, Wei Li, 2009, but differs in antennae placed the lower margin of eye, head exserted above the pronotal surface, lateral keels in prozona slightly contracted backward, upper margin of pronotum not waved in lateral view, width of tegmen 1.2 times width of mid leg femur and lower side of hind femur not black. The new species is also similar to Criotettix strivertex Zheng, Wei Li, 2009, it differs from latter by head exserted above the pronotal surface, upper margin of pronotum not waved in lateral view, lower side of mid femur straight, width of tegmen 1.2 times width of mid leg femur and length of hind femur 3.1 times its width. The type specimens are deposited in the College of Life Sciences, Hebei University, Baoding, China.

A new species and key to all known species of Confusacris Yin Li, 1987 (Orthoptera: Acrididae) from China.

A new species of the genus Confusacris Yin Li, 1987, namely i.e. Confusacris shanxiensis sp. nov. is described in this paper from Shanxi province of China. The new species is similar to Confusacris brachypterus Yin Li, 1987 but differs from the latter by tegmen of male extending over middle of hind femur distinctly, maximum width of cubital vein area 1.5 times median vein area, furculae large, epiphallus without acute projection on both sides and length of interspace of mesosternum1.6 times minimum width in female. A key to known species of the genus Confusacris Yin Li, 1987 is given in this paper. The type specimens are deposited in the College of Life Sciences, Hebei University, Baoding, China.

Application of DNA Barcoding in the Classification of Grasshoppers (Orthoptera: Acridoidea)-A Case Study of grasshoppers from Hebei Province, China.

Grasshoppers (Orthoptera: Acridoidea) are the main pests in agriculture, animal husbandry and forestry, and some species of grasshoppers can cause serious disaster. Taxonomy is the basis of pest control. Traditional morphological identification is time-consuming and laborious. It may be due to the existence of cryptic species or the limited number of morphologists, making the identification extremely unstable. In recent years, with the development of molecular systematics, DNA barcoding technology has been applied to environment, ecology, quarantine and so on. This study focuses on testing the feasibility of DNA barcoding in the species identification for superfamily Acridoidea. Sequences of the cox1 gene were obtained from 245 individuals of 43 species of Acridoidea and one species of Tetrigoidea as outgroup from Hebei Province. Phylogenetic, genetic distance and sequence difference threshold analyses using the Maximum Likelihood (ML), Automatic Barcode Gap Discovery (ABGD) and Molecular Defined Operational Taxonomic Units (MOTU) methods, respectively, were performed for obtained sequences and the 139 additional sequences of 21 species downloaded from GenBank. The results have shown that 40, 33, and 35 species among the 48 species are consistent with the traditional morphological classification based on the phylogenetic tree, ABGD and MOTU results, respectively and the DNA barcoding technology is very efficient and helpful for identifying the species of the superfamily Acridoidea; however, the morphological approach is still playing a key role in the species identifications. It also indicates that the cox1 gene is suitable for the phylogeny of genera and species level, but it is not suitable for the phylogenetic relationship of the advanced taxa such as families.

A new species and key to all known species of the genus Filchnerella Karny, 1908 from China (Orthoptera: Acridoidea, Pamphagidae).

The new species i.e. Filchnerella wuhaiensis sp. nov is described from Inner Mongolia Autonomous Region, China. The new species is similar to Filchnerella helanshanensis Zheng, 1992, but differs from the latter by 21-segmented antennae, prozona of pronotum higher than metazona, length of tegmina 2.0 times maximum width, width of interspace of mesosternum 1.2 times length and lower margin of hind femur not red in male. A key to all species of the genus Filchnerella Karny, 1908 is given in this paper. Two genera Pseudotmethis Bey-Bienko, 1948 and Paratmethis Zheng He, 1996 are valid genus and not a synonym of Filchnerella Karny, 1908. Type specimens are deposited in the College of Life Sciences, Hebei University, Baoding, Hebei, China.

Two new species of the genus Haplotropis Saussure, 1888 (Orthoptera, Acridoidea, Pamphagidae) from China.

Two new species of the genus Haplotropis Saussure, 1888 from China are described in this paper. The new species Haplotropis xiai sp. nov. is similar to Haplotropis brunneriana Saussure, 1888, but differs from latter by frontal ridge of male widened at median ocellus; tegmina narrower, cover 2/5 tympanum; cercus of male apical half part gently tapering; lower margin of epiphallus with high projection in the middle; anterior margin of pronotum in female with distinct acute angular in middle; length of subgenital plate shorter than width in female. The Haplotropis zhuoluensis sp. nov. is similar to Haplotropis xiai sp. nov., but differs from latter by anterior margin of pronotum reaching hind margin of eyes; length of temina is 1.6 times in male and 1.3 times in female of width; length of interspace shorter than narrowest in mesosternum of male; ancorae of epiphallus oblique inward distinctly, lower margin with high projection in the middle; length of subgenital plate longer than width in female. Type specimens are deposited in the College of Life Sciences, Hebei University, Baoding, China.

A review of the genus Eotmethis (Orthoptera, Acridoidea, Pamphagidae) with two new species and key to known species of the genus from China.

Two new species of the genus Eotmethis Bey-Bienko, 1948, namely Eotmethis inditibialis sp. nov. and Eotmethis shizuishanensis sp. nov. from the Ningxia Hui Autonomous Region are described in this paper. The new species Eotmethis inditibialis sp. nov. differs from all known species of the genus by inner side of hind tibia purple in the middle. The new species Eotmethis shizuishanensis sp. nov. is similar to Eotmethis xiai Ye, Shi & Zhi, 2015, differs from latter by tegmina of male shorter, reaching to 4th abdominal tergite, R vein without branch, M vein with one branch, Cu vein without branch and lower knee-lobe of hind femur yellow on inner side. The type specimens are deposited in College of Life Sciences, Hebei University, Baoding, China. Key to known species of the genus Eotmethis Bey-Bienko, 1948 is given.