PDIL1-2 can indirectly and negatively regulate expression of the AGPL1 gene in bread wheat.

BACKGROUND: ADP-glucose pyrophosphorylase (AGPase), the key enzyme in plant starch biosynthesis, is a heterotetramer composed of two identical large subunits and two identical small subunits. AGPase has plastidial and cytosolic isoforms in higher plants, whereas it is mainly detected in the cytosol of grain endosperms in cereal crops. Our previous results have shown that the expression of the TaAGPL1 gene, encoding the cytosolic large subunit of wheat AGPase, temporally coincides with the rate of starch accumulation and that its overexpression dramatically increases wheat AGPase activity and the rate of starch accumulation, suggesting an important role. METHODS: In this study, we performed yeast one-hybrid screening using the promoter of the TaAGPL1 gene as bait and a wheat grain cDNA library as prey to screen out the upstream regulators of TaAGPL1 gene. And the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method was used to verify the functional characterization of the identified regulators in starch biosynthesis. RESULTS: Disulfide isomerase 1-2 protein (TaPDIL1-2) was screened out, and its binding to the TaAGPL1-1D promoter was further verified using another yeast one-hybrid screen. Transiently silenced wheat plants of the TaPDIL1-2 gene were obtained by using BSMV-VIGS method under field conditions. In grains of BSMV-VIGS-TaPDIL1-2-silenced wheat plants, the TaAGPL1 gene transcription levels, grain starch contents, and 1000-kernel weight also significantly increased. CONCLUSIONS: As important chaperones involved in oxidative protein folding, PDIL proteins have been reported to form hetero-dimers with some transcription factors, and thus, our results suggested that TaPDIL1-2 protein could indirectly and negatively regulate the expression of the TaAGPL1 gene and function in starch biosynthesis.

Candidate Genes Expressed in Tolerant Common Wheat With Resistant to English Grain Aphid.

The English grain aphid, Sitobion avenae (F.) (Hemiptera: Aphididae), is a common worldwide pest of wheat (Triticum aestivum L.). The use of improved resistant cultivars by the farmers is the most effective and environmentally friendly method to control this aphid in the field. The winter wheat genotypes 98-10-35 and Amigo are resistant to S. avenae. To identify genes responsible for resistance to S. avenae in these genotypes, differential-display reverse transcription-polymerase chain reaction was used to identify the corresponding differentially expressed sequences in current study. Two backcross progenies were obtained by crossing the two resistant genotypes with the susceptible genotype 1376. Six potential expected-differential bands were sequenced. Lengths of the expressed sequence tags ranged from 128 to 532 bp. Although these expressed sequences were likely associated with S. avenae resistance, there was one expressed sequence tag located on 7DL chromosome, and its potential function may associate with the ability to maintain photosynthesis in wheat. That serves as an active way for tolerant common wheat with resistant to S. avenae. Cloning the full length of these sequences would help us thoroughly understand the mechanism of wheat resistance to S. avenae and be valuable for breeding cultivars with S. avenae resistance.

The 14-3-3 Protein BdGF14a Increases the Transcriptional Regulation Activity of BdbZIP62 to Confer Drought and Salt Resistance in Tobacco.

BdGF14a, a 14-3-3 gene from Brachypodium distachyon, induced by salt, H2O2, and abscisic acid (ABA), improved tolerance to drought and salt in tobacco, with a higher survival rate and longer roots under these stresses. Additionally, physiological index analyses showed that the heterologous expression of BdGF14a induced higher expression levels of antioxidant enzymes and their activities, leading to lighter DAB and NBT staining, denoting decreased H2O2 content. Additionally, the lower MDA content and ion leakage indicated enhanced cell membrane stability. Moreover, exogenous ABA resulted in shorter roots and a lower stomatal aperture in BdGF14a transgenic plants. BdGF14a interacted with NtABF2 and regulated the expression of stress-related genes. However, adding an ABA biosynthesis inhibitor suppressed most of these changes. Furthermore, similar salt and drought resistance phenotypes and physiological indicators were characterized in tobacco plants expressing BdbZIP62, an ABRE/ABF that interacts with BdGF14a. And Y1H and LUC assays showed that BdGF14a could enhance the transcription regulation activity of NtABF2 and BdbZIP62, targeting NtNECD1 by binding to the ABRE cis-element. Thus, BdGF14a confers resistance to drought and salinity through interaction with BdbZIP62 and enhances its transcriptional regulation activity via an ABA-mediated signaling pathway. Therefore, this work offers novel target genes for breeding salt- and drought-tolerant plants.

Expression of potential resistance genes to the English grain aphid, Sitobion avenae, in wheat, Triticum aestivum.

The English grain aphid, Sitobion avenae (F.) (Homoptera: Aphididae), is a dominant and destructive pest in wheat, Triticum estivum L. (Poales: Poaceae), production regions in China and other grain-growing areas worldwide. Patterns of gene expression of the S. avenae-resistant synthetic wheat line 98-10-35, the S. avenae-susceptible line1376, and their hybrid population, and the differences in segments between 98-10-35/1376 F3 resistant plants and resistant parents of 98-10-35, as well as those between the F3 resistant and susceptible populations, were examined with differential display reverse transcription PCR. The results showed that five patterns of differential expression were detected between the progeny and its resistant parents: 1) The gene was silenced in one of the parents; 2) Special expression showed in the progeny; 3) Expression was consistent with the resistant parents; 4) Up expression showed in the progeny but not in the parents; 5) Down expression showed in the progeny but not in the parents. Paired t-test results were not significant; however, the probability value (0.9158) indicated that gene expression on the RNA level were consistent with resistant bands found in F3 resistant individuals and resistant parents, as well as the F3 resistant and susceptible populations. For both the F3 of 98-10-35/1376 and the parents, the total number of amplified bands was 202, with an average of 25.3 per primer. The number of differential bands was 116, with an average of 14.5 per primer amplified and a polymorphism ratio of 56.3%. In the present study, differential expression genes in the resistant line 98-10-35 were all up-regulated. Among them, gene expression of resistant groups in the F3 population was in agreement with patterns 2, 3, and 4. However, the susceptible line 1376 did not have this gene expression on the RNA level. This pattern is expected to be used to select and analyze target genes from the same F3 population and the resistant parents. The results suggest that it can be employed as a new method for molecular assisted breeding.

Phylogeography and Genetic Structure of the Bush Cricket Decma fissa (Orthoptera, Tettigoniidae) in Southern China.

Decma fissa is the most widely distributed species of the genus Decma occuring in southern China. This study presents the first phylogeographic work of D. fissa based on COI, Cytb and ITS sequence. We examined genetic diversity with ITS and mitochondrial sequence respectively, and phylogenetic work was based on the mitochondrial data. A high-level genetic diversity was revealed based on mitochondrial data but a low-level diversity was shown with ITS sequence. For the mitochondrial data, divergence time analysis displayed five lineages. Based on the Mantel test, geographic and genetic distances among D. fissa populations revealed a significant positive correlation. Bayesian skyline plot (BSP) analyses implied that none of three major lineages of D. fissa was seemingly affected by the last glacial maximum (LGM, 0.015-0.025 Mya). Ecological niche modeling was used to predict the distribution of D. fissa in four periods (LGM, Mid-Holocene, current and 2070) in China. Analysis of the ancestral area reconstruction indicated that D. fissa occurred in the South China area.

A 14-3-3 Protein-Encoding Gene, BdGF14g, Confers Better Drought Tolerance by Regulating ABA Biosynthesis and Signaling.

Abscisic acid (ABA), a phytohormone, enacts a cardinal function in coping with abiotic stress. 14-3-3 proteins can interact with ABA-responsive-element-binding transcription factors (ABFs), a chief constituent of ABA signaling, and play critical roles in the dehydration response involving ABA signaling. Meanwhile, whether and how 14-3-3 proteins regulate ABA signaling to respond to aridity stress is yet to be fully investigated. Herein, BdGF14g, a 14-3-3 gene induced by ABA, H2O2, and PEG treatments, was identified in Brachypodium distachyon (B. distachyon). Overexpression of BdGF14g improved drought stress tolerance in tobacco plants, with a higher survival rate, longer root length, enhanced cell membrane stability, and increased antioxidase activity compared with non-transgenic controls in coping with dehydration. Both drought and exogenous ABA treatments resulted in smaller stomatal apertures in BdGF14g-transgenic lines. Additionally, when an ABA biosynthesis inhibitor was added, the better growth statuses, less H2O2 accumulation, and higher activities of catalase and superoxide dismutase under mannitol stress disappeared. Moreover, BdGF14g interacted with NtABF2, upregulated the endogenous ABA content, and enhanced the transcription of ABA-related genes, including NtNCED1, a crucial ABA biosynthesis gene, under drought conditions. In conclusion, BdGF14g acts as a positive factor in the water deficiency response by affecting ABA biosynthesis and signaling in tobacco plants.

Phylogeny and Integrative Taxonomy of the Genera Gymnaetoides and Pseudotachycines (Orthoptera: Rhaphidophoridae).

The genera Gymnaetoides and Pseudotachycines are endemic to China and are morphologically homogeneous. The few available diagnostic characters make species identification particularly challenging. Species cannot be classified according to the given generic diagnosis, and phylogenetic analyses have not been reported. Here, we reconstruct the phylogeny using Bayesian inference and maximum likelihood and employ four approaches to delimit species. The results suggest that both Gymnaetoides and Pseudotachycines are paraphyletic. Therefore, we revise their taxonomy based on the combination of morphological characters and molecular data. A new genus Homotachycines Zhu & Shi gen. nov. is erected, and six new combinations are proposed. Species delimitation identifies 15 new species and one new subspecies: Gymnaetoides huangshanensis, G. petalus, G. yangmingensis, G. lushanensis, Pseudotachycines procerus, P. procerus guizhouensis, P. zhengi, P. nephrus, P. sagittus, P. fengyangshanensis, Homotachycines triangulus, H. quadratus, H. baokangensis, H. fusus, H. concavus, and H. qinlingensis sp. nov. Moreover, we find that the shapes of the dorsal lateral lobes and the dorsal median lobe of the male genitalia are also important characters for identifying these genera and that the shapes of the dorsal and lateral sclerites of the male genitalia are suitable for the classifications of species.

Review of the genus Woznessenskia Gorochov, 2002 (Orthoptera: Gryllacrididae) from China.

The paper reports four new species of the genus Woznessenskia Gorochov, 2002 and gives supplementary information on Woznessenskia curvicauda(Bey-Bienko, 1962) from China. The specimens examined are deposited in the Museum of Hebei University, Baoding, China.

The Role of Feeding Characteristics in Shaping Gut Microbiota Composition and Function of Ensifera (Orthoptera).

Feeding habits were the primary factor affecting the gut bacterial communities in Ensifera. However, the interaction mechanism between the gut microbiota and feeding characteristics is not precisely understood. Here, the gut microbiota of Ensifera with diverse feeding habits was analyzed by shotgun metagenomic sequencing to further clarify the composition and function of the gut microbiota and its relationship with feeding characteristics. Our results indicate that under the influence of feeding habits, the gut microbial communities of Ensifera showed specific characteristics. Firstly, the gut microbial communities of the Ensifera with different feeding habits differed significantly, among which the gut microbial diversity of the herbivorous Mecopoda niponensis was the highest. Secondly, the functional genes related to feeding habits were in high abundance. Thirdly, the specific function of the gut microbial species in the omnivorous Gryllotalpa orientalis showed that the more diverse the feeding behavior of Ensifera, the worse the functional specificity related to the feeding characteristics of its gut microbiota. However, feeding habits were not the only factors affecting the gut microbiota of Ensifera. Some microorganisms' genes, whose functions were unrelated to feeding characteristics but were relevant to energy acquisition and nutrient absorption, were detected in high abundance. Our results were the first to report on the composition and function of the gut microbiota of Ensifera based on shotgun metagenomic sequencing and to explore the potential mechanism of the gut microbiota's association with diverse feeding habits.

Further notes on the genus Ocellarnaca Gorochov, 2004 (Orthoptera: Gryllacrididae) from China.

The paper reports three new species of the genus Ocellarnaca Gorochov, 2004 (Orthoptera: Gryllacrididae) from China, i.e. Ocellarnaca brachyptera sp. nov., Ocellarnaca nigrofemora sp. nov., Ocellarnaca nigra sp. nov. The type specimens are deposited in the Museum of Hebei University, Baoding 071002, China.

Gut bacterial communities across 12 Ensifera (Orthoptera) at different feeding habits and its prediction for the insect with contrasting feeding habits.

Insect microbial symbioses play a critical role in insect lifecycle, and insect gut microbiome could be influenced by many factors. Studies have shown that host diet and taxonomy have a strong influence on insect gut microbial community. In this study, we performed sequencing of V3-V4 region of 16S rRNA gene to compare the composition and diversity of 12 Ensifera from 6 provinces of China. Moreover, the influences of feeding habits and taxonomic status of insects on their gut bacterial community were evaluated, which might provide reference for further application research. The results showed that Proteobacteria (45.66%), Firmicutes (34.25%) and Cyanobacteria (7.7%) were the predominant bacterial phyla in Ensifera. Moreover, the gut bacterial community composition of samples with different feeding habits was significantly different, which was irrespective of their taxa. The highest diversity of gut bacteria was found in the omnivorous Ensifera. Furthermore, common and unique bacteria with biomarkers were found based on the dietary characteristics of the samples. However, the bacterial community structure of the Ensifera samples was significantly different from that of Caelifera. Therefore, we concluded that feeding habits and taxonomic status jointly affect the gut bacterial community composition of the samples from Orthoptera. However, the influence of feeding habit dominates when taxonomy category below the suborder level. In addition, the dominant, common and unique bacterial community structure could be used to predict the contrastic feeding habits of insects belonging to Ensifera.

Remarks on the genus Tachycines Adelung, 1902 (Orthoptera: Rhaphidophoridae: Aemodogryllinae) with description of eight new species from caves in southern China.

This paper describes eight new cave-dwelling species of the genus Tachycines from Guizhou, Hunan, Guangxi and Sichuan, i.e. Tachycines (Gymnaeta) liboensis sp. nov., Tachycines (Gymnaeta) bifolius sp. nov., Tachycines (Gymnaeta) tuberus sp. nov., Tachycines (Gymnaeta) paradoxus sp. nov., Tachycines (Gymnaeta) umbellus sp. nov., Tachycines (Gymnaeta) nulliscleritus sp. nov., Tachycines (Gymnaeta) taenus sp. nov. and Tachycines (Gymnaeta) latiliconcavus sp. nov. The females of T. (G.) ferecaecus (Gorochov, Rampini Di Russo, 2006) and T. (G.) proximus (Gorochov, Rampini Di Russo, 2006) are first recorded. Moreover, we consider T. (G.) aspes (Rampini Di Russo, 2008) as a valid species instead of a synonym of T. (G.) proximus (Gorochov, Rampini Di Russo, 2006) according to obviously different reduction of fastigium verticis and eyes, higher number of spines on the hind tibia and the shape of male genitalia. Furthermore, Eutachycines crenatus (Gorochov, Rampini Di Russo, 2006) should be retransferred to the subgenus Tachycines (Gymnaeta) due to the diagnosis of genus Eutachycines: dorsal sclerite of male genitalia with a protuberance in the upper part. Therefore, it should still belong to the subgenus Tachycines (Gymnaeta). In addition, morphological photographs of the eight new species and five known species are provided, including eyeless species.

Supplement of the genus Diestramima Storozhenko, 1990 (Orthoptera: Rhaphidophoridae: Aemodogryllinae) from China.

This paper describes two new species of the genus Diestramima from China, i.e. Diestramima arbora sp. nov. and Diestramima lamina sp. nov. The morphological photographs of the two species are provided. All specimens are preserved in the Museum of Hebei University.

Review of the genus Diestramima Storozhenko, 1990 (Orthoptera: Rhaphidophoridae: Aemodogryllinae) from China.

In this paper, 6 new species, 1 new subspecies and 1 newly recorded species of the genus Diestramima from China are described, i.e. Diestramima acutiapicis sp. nov., Diestramima bina sp. nov., Diestramima cycla sp. nov., Diestramima truncata sp. nov., Diestramima subtilis sp. nov., Diestramima eurya sp. nov., Diestramima austrosinensis quaterna ssp. nov. and Diestramima major Gorochov, 1998. The morphological photographs of these and 8 known species are provided. All specimens examined are preserved in the Museum of Hebei University.

Remarks on the genus Tamdaotettix Gorochov, 1998 (Orthoptera: Rhaphidophoridae: Aemodogryllinae) from China.

In this paper, a new species of the genus Tamdaotettix from China, Tamdaotettix (Tamdaotettix) longituberus sp. nov., is described, with a new recorded species, Tamdaotettix (Tamdaotettix) dilutus Gorochov, 1998 from China. The morphological photographs of Tamdaotettix (Tamdaotettix) longituberus sp. nov., Tamdaotettix (Tamdaotettix) dilutus Gorochov, 1998, Tamdaotettix (Tamdaotettix) tridenticulatus Qin, Liu Li, 2016 and the distribution data of the genus Tamdaotettix from China are provided. All specimens examined are preserved in the Museum of Hebei University.

The genus Mimadiestra Storozhenko Dawwrueng, 2014 (Orthoptera: Rhaphidophoridae) from China, with description of two new species.

In this paper, the genus Mimadiestra Storozhenko Dawwrueng, 2014 is reported from China and two new species are described, i.e. Mimadiestra gracila sp. nov. and Mimadiestra dicha sp. nov. Morphological photographs of the two species are provided. All type specimens are preserved in the Museum of Hebei University.

One new species and one new recorded species of the genus Kuzicus (Tettigoniidae: Meconematinae: Meconematini) from China.

In this paper, a new species of the genus Kuzicus from China, Kuzicus (Kuzicus) leptocercus sp. nov., is described, with a new recorded species, Kuzicus (Kuzicus) koeppeli Sänger & Helfert, 2004 from China. The distribution data and morphological photographs of Kuzicus (Kuzicus) leptocercus sp. nov. and Kuzicus (Kuzicus) koeppeli Sänger & Helfert, 2004 are provided. All specimens are preserved in the Museum of Hebei University.

De Novo Assembly and Transcriptome Analysis of Wheat with Male Sterility Induced by the Chemical Hybridizing Agent SQ-1.

Wheat (Triticum aestivum L.), one of the world's most important food crops, is a strictly autogamous (self-pollinating) species with exclusively perfect flowers. Male sterility induced by chemical hybridizing agents has increasingly attracted attention as a tool for hybrid seed production in wheat; however, the molecular mechanisms of male sterility induced by the agent SQ-1 remain poorly understood due to limited whole transcriptome data. Therefore, a comparative analysis of wheat anther transcriptomes for male fertile wheat and SQ-1-induced male sterile wheat was carried out using next-generation sequencing technology. In all, 42,634,123 sequence reads were generated and were assembled into 82,356 high-quality unigenes with an average length of 724 bp. Of these, 1,088 unigenes were significantly differentially expressed in the fertile and sterile wheat anthers, including 643 up-regulated unigenes and 445 down-regulated unigenes. The differentially expressed unigenes with functional annotations were mapped onto 60 pathways using the Kyoto Encyclopedia of Genes and Genomes database. They were mainly involved in coding for the components of ribosomes, photosynthesis, respiration, purine and pyrimidine metabolism, amino acid metabolism, glutathione metabolism, RNA transport and signal transduction, reactive oxygen species metabolism, mRNA surveillance pathways, protein processing in the endoplasmic reticulum, protein export, and ubiquitin-mediated proteolysis. This study is the first to provide a systematic overview comparing wheat anther transcriptomes of male fertile wheat with those of SQ-1-induced male sterile wheat and is a valuable source of data for future research in SQ-1-induced wheat male sterility.

PDIL1-2 can indirectly and negatively regulate expression of the AGPL1 gene in bread wheat

BACKGROUND: ADP-glucose pyrophosphorylase (AGPase), the key enzyme in plant starch biosynthesis, is a heterotetramer composed of two identical large subunits and two identical small subunits. AGPase has plastidial and cytosolic isoforms in higher plants, whereas it is mainly detected in the cytosol of grain endosperms in cereal crops. Our previous results have shown that the expression of the TaAGPL1 gene, encoding the cytosolic large subunit of wheat AGPase, temporally coincides with the rate of starch accumulation and that its overexpression dramatically increases wheat AGPase activity and the rate of starch accumulation, suggesting an important role. METHODS: In this study, we performed yeast one-hybrid screening using the promoter of the TaAGPL1 gene as bait and a wheat grain cDNA library as prey to screen out the upstream regulators of TaAGPL1 gene. And the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method was used to verify the functional characterization of the identified regulators in starch biosynthesis. RESULTS: Disulfide isomerase 1-2 protein (TaPDIL1-2) was screened out, and its binding to the TaAGPL1-1D promoter was further verified using another yeast one-hybrid screen. Transiently silenced wheat plants of the TaPDIL1-2 gene were obtained by using BSMV-VIGS method under field conditions. In grains of BSMV-VIGS-TaPDIL1-2-silenced wheat plants, the TaAGPL1 gene transcription levels, grain starch contents, and 1000-kernel weight also significantly increased. CONCLUSIONS: As important chaperones involved in oxidative protein folding, PDIL proteins have been reported to form hetero-dimers with some transcription factors, and thus, our results suggested that TaPDIL1-2 protein could indirectly and negatively regulate the expression of the TaAGPL1 gene and function in starch biosynthesis.

[Analysis of three wheat cytoplasmic male sterile lines mitochondrial DNA by AFLP].

Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.