Isoform X1 Research Articles

Obstructor, a Frankliniella occidentalis protein, promotes transmission of tomato spotted wilt orthotospovirus.

Tomato spotted wilt orthotospovirus (TSWV) causes substantial economic losses to vegetables and other crops. TSWV is mainly transmitted by thrips in a persistent and proliferative manner, and its most efficient vector is the western flower thrips, Frankliniella occidentalis (Pergande). In moving from the thrips midgut to the salivary glands in preparation for transmission, the virions must overcome multiple barriers. Although several proteins that interact with TSWV in thrips have been characterized, we hypothesized that additional thrips proteins interact with TSWV and facilitate its transmission. In the current study, 67 F.occidentalis proteins that interact with GN (a structural glycoprotein) were identified using a split-ubiquitin membrane-based yeast 2-hybrid (MbY2H) system. Three proteins, apolipoprotein-D (ApoD), orai-2-like (Orai), and obstructor-E-like isoform X2 (Obst), were selected for further study based on their high abundance and interaction strength; their interactions with GN were confirmed by MbY2H, yeast β-galactosidase and luciferase complementation assays. The relative expressions of ApoD and Orai were significantly down-regulated but that of Obst was significantly up-regulated in viruliferous thrips. When interfering with Obst in larval stage, the TSWV acquisition rate in 3 independent experiments was significantly decreased by 26%, 40%, and 35%, respectively. In addition, when Obst was silenced in adults, the virus titer was significantly decreased, and the TSWV transmission rate decreased from 66.7% to 31.9% using the leaf disk method and from 86.67% to 43.33% using the living plant method. However, the TSWV acquisition and transmission rates were not affected by interference with the ApoD or Orai gene. The results indicate that Obst may play an important role in TSWV acquisition and transmission in Frankliniella occidentalis.

Effect of dietary carbohydrate on glycometabolism of juvenile golden pompano (Trachinotus ovatus) related to growth performance, hepatic histopathology, transcriptome profiles and identification of differentially expressed genes

A hepatic transcriptome was produced through the technology of RNA sequencing and short-read assembly in golden pompano Trachinotus ovatus fed with three experimental diets respectively containing low carbohydrate diet (LCD), intermediate carbohydrate diet (MCD), and high carbohydrate diet (HCD). In total, 57,217 unigenes were acquired in the current study, with average length 1182.24 bp. 21,920, 22,514 and 24,680 unigenes were acquired from LCD, MCD and HCD groups, respectively. There were 1513 differential expressed genes in LCD and MCD groups, with 676 up- and 837 down-regulated genes in LCD group; 2369 genes were differentially expressed between HCD vs MCD, containing 1397 up and 972 down-regulated genes in HCD group. 1686 differential expressed genes were observed in the comparison of HCD and LCD, containing 1044 up- and 642 down-regulated genes in HCD group. Some differential expressed genes, such as hexokinase-1-like isoform X3, pancreatic alpha-amylase-like, hexokinase-1-like isoform X1, ADP-dependent glucokinase isoform X2, fructose-bisphosphate aldolase A-like, and acetyl-CoA carboxylase isoform X8, were related to carbohydrate metabolism, indicating these genes in the golden pompano were in response to dietary carbohydrate. In this study, long-chain-fatty-acid-CoA ligase 5 was up-regulated in comparison of HCD_vs_MCD and HCD_vs_LCD, which indicated that FACL5 may be sensitive to high dietary carbohydrate. This result coincides with the pathological changes of hepatocyte morphology in the present study. The results of transcriptome analysis in this study were confirmed by RT-PCR in 14 randomly selected unigenes. This study may provide novel data and theoretical support on hepatic transcriptome information of the golden pompano, which is a vital resource for studies of bioinformatics in golden pompano. Besides, the present study identifies some pivotal genes related to dietary carbohydrate and lipid deposition, which will accumulate data on carbohydrate and/or lipid metabolism in marine fish species to support related research.

Distinct proteomic profile of ovarian follicular fluid in ewes from small versus large developing follicles

This study aimed to describe the proteins of the ovarian follicular fluid from ewes. The follicular fluid was taken from 294 ovaries collected from slaughterhouses and split into two groups based on follicle size: Group 1 (< 4 mm); and Group 2 (≥ 4 to 6 mm). The samples for ESI-Q-TOF MS/MS mass spectrometry were prepared through Tryptic in-gel digestion. For comparisons between groups, the data were analyzed using uni and multivariate statistical analysis, represented by principal component analysis (PCA), the measure of a variable's importance (VIP score) in the partial least squares-discriminant analysis (PLS-DA), the fold change, and t-test, using the exponentially modified protein abundance index (emPAI). Moreover, gene ontology was performed to identify molecular function, biological process, and cellular component. Fifty-five major proteins were identified in the 2 groups. The separation of the formed clusters was clearly observed in the PCA. Twelve proteins had a VIP score with α ≥ 1.5. Only 2 proteins were found in higher abundance in Group 1, the serum amyloid A-4 protein isoform X1 and trinucleotide repeat-containing gene 6c protein isoform X2. However, in Group 2, 10 proteins were in higher abundance: complement component C6, serotransferrin isoform X2, fibrinopeptide A, tetranectin, inter-alpha-trypsin inhibitor heavy chain H4 isoform X1, serpin B9 isoform X2, alpha-enolase isoform X6, serum paraoxonase/arylesterase 1, complement factor B, and complement C3 isoform X2, partial. We concluded, therefore, that a distinct profile of protein abundance is observed in small vs large growing follicles, including proteins especially related to ovulation, follicular metabolism, and oocyte development and quality.

Identification and Characterization of the Larval Settlement Pheromone Protein Components in Adult Shells of Crassostrea gigas: A Novel Function of Shell Matrix Proteins

The global decline of natural oyster populations emphasizes the need to improve our understanding of their biology. Understanding the role of chemical cues from conspecifics on how oysters occupy appropriate substrata is crucial to learning about their evolution, population dynamics, and chemical communication. Here, a novel role of a macromolecular assembly of shell matrix proteins which act as Crassostrea gigas Settlement Pheromone Protein Components in adult shells is demonstrated as the biological cue responsible for gregarious settlement on conspecifics. A bioassay-guided fractionation approach aided by biochemical and molecular analyses reveals that Gigasin-6 isoform X1 and/or X2 isolated from adult shells is the major inducing cue for larval settlement and may also play a role in postlarva–larva settlement interactions. Other isolated Stains-all-stainable acidic proteins may function as a co-factor and a scaffold/structural framework for other matrix proteins to anchor within this assembly and provide protection. Notably, conspecific cue-mediated larval settlement induction in C. gigas presents a complex system that requires an interplay of different glycans, disulfide bonds, amino acid groups, and phosphorylation crosstalk for recognition. These results may find application in the development of oyster aquacultures which could help recover declining marine species and as targets of anti-fouling agents.

Transcriptomic response of intertidal brittle star Ophiothrix exigua to seasonal variation.

Adaptation to seasonal change is essential for survival, and is especially critical for organisms living in physically harsh environments. Brittle stars (Ophiothrix), known as a keystone species, inhabiting the intertidal rocky ecosystem are affected by multiple stressors, but molecular insights into their adaptation remain poorly studied. In the present study, transcriptomic responses of Ophiothrix exigua from the intertidal habitats of the North Pacific Ocean during summer and winter are reported. A total of 12,844 differentially expressed genes (DEGs) were identified. Of these, 7102 genes were up-regulated and 5742 genes were down-regulated in summer relative to winter. One hundred fifty-two key DEGs, including 31 up-regulated and 121 down-regulated genes, were categorized into three major subcategories and seven subclasses. The key DEGs included heat shock cognate protein 70 (HSC70), toll-like receptor-2 (TLR2), cAMP-dependent protein kinase catalytic subunit beta-like isoform X2 (PKA), serine/threonine-protein kinase mTOR (MTOR), and ras-related c3 botulinum toxin substrate 1 isoform X1 (RAC1). Glutathione peroxidase-like (GPX) and tubulin superfamily members (TUBA, TUBB) were consistent across seasons. The main defense-related pathways in brittle star were phagosome, apoptosis, and glutathione metabolism. These findings would greatly enhance our understanding of the genomic basis of environmental adaptation in intertidal invertebrates.

Proteomic and parallel reaction monitoring approaches to evaluate biomarkers of mutton tenderness

Intensive fattening usually results in the changes of meat quality. Tenderness is a central attribute for mutton sensory qualities and consumers’ choice. Here, we reported that intensive fattening mutton was more tender than that of traditionally raised sheep. By proteomic approach, we found 49 differentially expressed proteins in longissimus dorsi muscle. After bioinformatics analysis, 5 cytoskeletal proteins, 3 protein binding proteins and 7 metabolic enzymes were identified as potential biomarkers for mutton tenderness. Finally, we verified the expression of these abundant proteins by parallel reaction monitoring (PRM). Collectively, our results reveal that the mutton of sheep raised by intensive fattening is more tender than that of traditionally raised sheep. Myosin-2, myosin-13, vimentin, carbonic anhydrase, carbonic anhydrase-2, Glutathione S-transferase and Microtubule-associated protein 4 isoform X1 can be candidate biomarkers for mutton tenderness. Our data also indicate a central role of cytoskeletal proteins and metabolic enzymes in determining mutton tenderness.

Full-Length Transcriptome Reconstruction Reveals the Genetic Mechanisms of Eyestalk Displacement and Its Potential Implications on the Interspecific Hybrid Crab (Scylla serrata ♀ × S. paramamosain ♂).

Simple SummaryThe eyestalk is a key organ in crustaceans that produces neurohormones and regulates a range of physiological functions. Eyestalk displacement was discovered in some first-generation (F1) offspring of the novel interspecific hybrid crab (Scylla serrata ♀ × S. paramamosain ♂). To uncover the genetic mechanism underlying eyestalk displacement and its potential implications, high-quality transcriptome was reconstructed using single-molecule real-time (SMRT) sequencing. A total of 37 significantly differential alternative splicing (DAS) events (17 up-regulated and 20 down-regulated) and 1475 significantly differential expressed transcripts (DETs) (492 up-regulated and 983 down-regulated) were detected in hybrid crabs with displaced eyestalks (DH). The most significant DAS events and DETs were annotated as being endoplasmic reticulum chaperone BiP and leucine-rich repeat protein lrrA-like isoform X2. In addition, the top ten significant gene ontology (GO) terms were related to the cuticle or chitin. Overall, this study highlights the underlying genetic mechanisms of eyestalk displacement and provide useful knowledge for mud crab (Scylla spp.) crossbreeding.The lack of high-quality juvenile crabs is the greatest impediment to the growth of the mud crab (Scylla paramamosain) industry. To obtain high-quality hybrid offspring, a novel hybrid mud crab (S. serrata ♀ × S. paramamosain ♂) was successfully produced in our previous study. Meanwhile, an interesting phenomenon was discovered, that some first-generation (F1) hybrid offspring’s eyestalks were displaced during the crablet stage I. To uncover the genetic mechanism underlying eyestalk displacement and its potential implications, both single-molecule real-time (SMRT) and Illumina RNA sequencing were implemented. Using a two-step collapsing strategy, three high-quality reconstructed transcriptomes were obtained from purebred mud crabs (S. paramamosain) with normal eyestalks (SPA), hybrid crabs with normal eyestalks (NH), and hybrid crabs with displaced eyestalks (DH). In total, 37 significantly differential alternative splicing (DAS) events (17 up-regulated and 20 down-regulated) and 1475 significantly differential expressed transcripts (DETs) (492 up-regulated and 983 down-regulated) were detected in DH. The most significant DAS events and DETs were annotated as being endoplasmic reticulum chaperone BiP and leucine-rich repeat protein lrrA-like isoform X2. In addition, the top ten significant GO terms were related to the cuticle or chitin. Overall, high-quality reconstructed transcriptomes were obtained for the novel interspecific hybrid crab and provided valuable insights into the genetic mechanisms of eyestalk displacement in mud crab (Scylla spp.) crossbreeding.

Phycoremediation of Copper by Chlorella protothecoides (UTEX 256): Proteomics of Protein Biosynthesis and Stress Response

Phycoremediation is an eco-friendly treatment for mining wastes. Copper at high concentrations is toxic for microalgae growth (bioremediation). Proteomics is a modern approach that can assist in elucidating, in detail, the highly complex metabolic mechanisms related to phycoremediation. Therefore, this study aimed to evaluate the effect of copper ions (Cu2+) on the metabolism of Chlorella protothecoides (UTEX 256), particularly the proteome changes. The WC culture medium supplemented with Cu2+ at 0.3, 0.6, and 0.9 mg/L showed a strict correlation to Cu2+ removal of 40, 33, and 36% of the initial content, respectively. In addition, Cu2+ concentrations did not affect microalgae growth—a very traditional approach to measuring toxicity. However, the proteomics data indicated that when compared to the control, reductions in protein levels were observed, and the 10 most scored proteins were related to the light-harvesting complex. Interestingly, C. protothecoides cultivated at 0.9 mg of Cu2+/L biosynthesized the protein Ycf3-interacting chloroplastic isoform X1 to respond to the photooxidative stress and the DNA-directed RNA polymerase III subunit RPC5 was related to the Cu2+ binding. Pre-mRNA-processing factor 19 and cytochrome c peroxidase proteins were observed only in the copper-containing treatments indicating the activation of antioxidant mechanisms by reactive oxygen species, which are potential environmental pollutant biomarkers.

The Role of Intestinal Microbial Metabolites in the Immunity of Equine Animals Infected With Horse Botflies.

The microbiota and its metabolites play an important role in regulating the host metabolism and immunity. However, the underlying mechanism is still not well studied. Thus, we conducted the LC-MS/MS analysis and RNA-seq analysis on Equus przewalskii with and without horse botfly infestation to determine the metabolites produced by intestinal microbiota in feces and differentially expressed genes (DEGs) related to the immune response in blood and attempted to link them together. The results showed that parasite infection could change the composition of microbial metabolites. These identified metabolites could be divided into six categories, including compounds with biological roles, bioactive peptides, endocrine-disrupting compounds, pesticides, phytochemical compounds, and lipids. The three pathways involving most metabolites were lipid metabolism, amino acid metabolism, and biosynthesis of other secondary metabolites. The significant differences between the host with and without parasites were shown in 31 metabolites with known functions, which were related to physiological activities of the host. For the gene analysis, we found that parasite infection could alarm the host immune response. The gene of “cathepsin W” involved in innate and adaptive immune responses was upregulated. The two genes of the following functions were downregulated: “protein S100-A8” and “protein S100-A9-like isoform X2” involved in chemokine and cytokine production, the toll-like receptor signaling pathway, and immune and inflammatory responses. GO and KEGG analyses showed that immune-related functions of defense response and Th17 cell differentiation had significant differences between the host with and without parasites, respectively. Last, the relationship between metabolites and genes was determined in this study. The purine metabolism and pyrimidine metabolism contained the most altered metabolites and DEGs, which mainly influenced the conversion of ATP, ADP, AMP, GTP, GMP, GDP, UTP, UDP, UMP, dTTP, dTDP, dTMP, and RNA. Thus, it could be concluded that parasitic infection can change the intestinal microbial metabolic activity and enhance immune response of the host through the pathway of purine and pyrimidine metabolism. This results will be a valuable contribution to understanding the bidirectional association of the parasite, intestinal microbiota, and host.

Pharmacophore identification, virtual screening and activity verification of pedunsaponin A on target proteins PcAdv and PcnWAS of Pomacea canaliculata.

Target-protein-based pesticide screening has attracted wide-ranging attention on pesticide science. Pedunsaponin A (PA) is a compound isolated from the root of Pueraria peduncularis, and it has a strong toxic effect on Pomacea canaliculata. Previous studies found that Advlin (PcAdv) and neural Wiskott-Aldrich syndrome isoform X1(PcnWAS) are target proteins of PA when interacted with P.canaliculata. In this study, we modeled the two target proteins through I-Tasser and identified the pharmacophore of PA binding to the two target proteins by molecular docking. Furthermore, through virtual screening, potassium alginate was found to strongly bind to the target proteins in theory. In vivo bioassay showed that, similar to PA treatment, potassium alginate was able to induce typical poisoning symptoms on P.canaliculata, which were characterized by abnormal increase of excreta, weakening of climbing capacity, loss of gill cilia and decrease in hemocyanin content, and even cause death of P.canaliculata with a 13.33% mortality rate under 100 mg L-1 concentration. Furthermore, the treatment of potassium alginate also decreased the gene expression level of PcAdv and PcnWAS. These findings indicate that potassium alginate can affect the living state of P.canaliculata, and that it is feasible to develop new molluscicides based on PcAdv and PcnWAS by virtual screening. © 2022 Society of Chemical Industry.

Spatial Expression Patterning of Kelch-Like Protein Homolog 10 (klhl10), Armadillo Repeat-Containing Protein 4 Isoform X2 (armc4), and a Gamete-Specific Mitochondrial Cytochrome C Oxidase (mt-co1) during Spermatogenesis/Spermiogenesis in the Mantle of Male Blue Mussel (Mytilus edulis)

The spatial expression patterns of three genes [kelch-like protein homolog 10 (klhl10); armadillo repeat-containing protein 4 isoform X2 (armc4); mitochondria cytochrome oxidase I (mt-co1)] known to be important mechanistic components of spermatogenesis/spermiogenesis were localized in mantle tissue sections of male Mytilus edulis using nonisotopic in situ hybridization and RNA probes based on sequences derived from available mantle RNA sequencing libraries. The expression patterns showed that all three genes are primarily localized in a layer of cells adjacent to the outer wall of the testicular acini. This layer is known to contain developing stages of spermatogonia, spermatocytes, and spermatids. Kelch-like protein homolog 10 was expressed in immature spermatogonial cells/spermatocytes characterized by large round central nuclei and situated near the periphery of the testicular acini with little to no expression in adjacent populations of spermatids or mature spermatozoa at the center of the acini. Like klhl10, armc4 transcripts were also detected in the cells near the inner periphery of the outer acinar wall. In contrast, however, the armc4 expressing cells appeared to be spatially shifted more toward the inner edge of the cell layer and toward the center of the acini. The localization of cells expressing armc4 transcripts near the inner edge of the developing sperm layer in the testicular acini of M. edulis supports a role for armc4 in spermatid morphogenesis and maturation. Mitochondria-specific cytochrome c oxidase transcripts were localized in cells throughout the inner periphery of the testicular acini wall but not in association with mature spermatozoa in the center of the acini reflecting the significant level of cellular activity that occurs during early sperm development. This supports an important mitochondrial role in early and mid-spermatogenesis in M. edulis potentially related to sperm quality control. This work provides a first view into the molecular mechanisms behind spermatogenesis/spermiogenesis in M. edulis.

New Insight on Vitality Differences for the Penaeid Shrimp, Fenneropenaeus chinensis, in Low Salinity Environment Through Transcriptomics

Excessive rainfall changes salinity in shrimp farming ponds in short period and exerts low salinity stress on the outdoor breeding shrimp under global warming. Fenneropenaeus chinensis can have different performance on vitality in low salinity environments. To reveal mechanisms of vitality difference in shrimp living in low saline environments. This study based on the normal and moribund F. chinensis in 10 ppt salinity environment using high-throughput sequencing identifies 1,429 differentially expressed genes (DEGs), 586 of which are upregulated, while 843 of which are downregulated in the normal group (FCN10) as compared to the moribund group (FCM10). Meanwhile, another transcriptomic analysis is conducted on the normal and moribund shrimp from 25 ppt (FCN25 vs. FCM25) salinity environment as the control, in which 1,311 DEGs (upregulated: 327 genes, downregulated: 984 genes) are identified. In this study, intersective pathways, GO (Gene Ontology) categories and DEGs from the two groups of comparative transcriptome are investigated. The two intersective pathways (Metabolism of xenobiotics by cytochrome P450, Pentose, and glucuronate interconversions) significantly enriched by DEGs are related to detoxification. In these two pathways, there is one vitality regulation-related gene (VRRG), the Dhdh (dihydrodiol dehydrogenase), which is upregulated in both the groups of FCN10 and FCN25 as compared to the groups of FCM10 and FCM25, respectively. Similarly, in the 25 top intersective GO categories, four VRRGs are revealed. Three of them are upregulated (Itgbl, kielin/chordin-like protein, Slc2a8, solute carrier family 2, facilitated glucose transporter member 8-like protein and Cyp3a30, cytochrome P450 3A30-like protein); one of them is downregulated (Slc6a9, sodium-dependent nutrient amino acid transporter 1-like protein isoform X2). These GO categories are related to transmembrane transporter activity of substance, enzyme inhibitor activity, monooxygenase activity. RT-qPCR analysis further verifies the VRRGs. The study gives new insight into understanding the vitality differences for F. chinensis, in low salinity environment. The pathways and DEGs in response to low salinity stress in modulating the vitality of F. chinensis that could serve as tools in future genetic studies and molecular breeding.

Overexpression of the Salix matsudana SmAP2-17 gene improves Arabidopsis salinity tolerance by enhancing the expression of SOS3 and ABI5

BackgroundSalix matsudana (Koidz.) is a widely planted ornamental allotetraploid tree species. Genetic engineering can be used to enhance the tolerance of this species to soil salinization, endowing varieties with the ability to grow along coastlines, thereby mitigating afforestation and protecting the environment. The AP2/ERF family of transcription factors (TFs) plays multidimensional roles in plant biotic/abiotic stress tolerance and plant development. In this study, we cloned the SmAP2-17 gene and performed functional analysis of its role in salt tolerance. This study aims to identify key genes for future breeding of stress-resistant varieties of Salix matsudana.ResultsSmAP2-17 was predicted to be a homolog of AP2-like ethylene-responsive transcription factor ANT isoform X2 from Arabidopsis, with a predicted ORF of 2058 bp encoding an estimated protein of 685 amino acids containing two conserved AP2 domains (PF00847.20). SmAP2-17 had a constitutive expression pattern and was localized to the nucleus. The overexpression of the native SmAP2-17 CDS sequence in Arabidopsis did not increase salt tolerance because of the reduced expression level of ectopic SmAP2-17, potentially caused by salt-induced RNAi. Transgenic lines with high expression of optimized SmAP2-17 CDS under salt stress showed enhanced tolerance to salt. Moreover, the expression of general stress marker genes and important salt stress signaling genes, including RD29A, ABI5, SOS3, AtHKT1, and RBohF, were upregulated in SmAP2-17-overexpressed lines, with expression levels consistent with that of SmAP2-17 or optimized SmAP2-17. Promoter activity analysis using dual luciferase analysis showed that SmAP2-17 could bind the promoters of SOS3 and ABI5 to activate their expression, which plays a key role in regulating salt tolerance.ConclusionsThe SmAP2-17 gene isolated from Salix matsudana (Koidz.) is a positive regulator that improves the resistance of transgenic plants to salt stress by upregulating SOS3 and ABI5 genes. This study provides a potential functional gene resource for future generation of salt-resistant Salix lines by genetic engineering.

Synovial fluid proteome profile of surgical versus chemical induced osteoarthritis in rabbits.

BackgroundAnimal models are significant for understanding human osteoarthritis (OA). This study compared the synovial fluid proteomics changes in surgical and chemical induced OA models.MethodsThirty rabbits either had anterior cruciate ligament transection (ACLT) procedure or injected intra-articularly with monosodium iodoacetate (MIA, 8 mg) into the right knee. The joints were anatomically assessed, and the synovial fluid proteins analyzed using two-dimensional polyacrylamide gel electrophoresis (2DGE) and MALDI TOF/TOF mass spectrometry analysis at 4, 8 and 12 weeks. The proteins’ upregulation and downregulation were compared with control healthy knees.ResultsSeven proteins (histidine-rich glycoprotein, beta-actin-like protein 2 isoform X1, retinol-binding protein-4, alpha-1-antiproteinase, gelsolin isoform, serotransferrin, immunoglobulin kappa-b4 chain-C-region) were significantly expressed by the surgical induction. They characterized cellular process (27%), organization of cellular components or biogenesis (27%), localization (27%) and biological regulation (18%), which related to synovitis, increased cellularity, and subsequently cartilage damage. Three proteins (apolipoprotein I-IV precursor, serpin peptidase inhibitor and haptoglobin precursor) were significantly modified by the chemical induction. They characterized stimulus responses (23%), immune responses (15%), biological regulations (15%), metabolism (15%), organization of cellular components or biogenesis (8%), cellular process (8%), biological adhesions (8%) and localization (8%), which related to chondrocytes glycolysis/death, neovascularization, subchondral bone necrosis/collapse and inflammation.ConclusionsThe surgical induced OA model showed a wider range of protein changes, which were most upregulated at week 12. The biological process proteins expressions showed the chemical induced joints had slower OA progression compared to surgical induced joints. The chemical induced OA joints showed early inflammatory changes, which later decreased.

ITRAQ Proteomic Analysis of Interactions Between 20E and Phospholipase C in Apolygus lucorum (Meyer-Dür).

Polyphagous Apolygus lucorum has become the dominant insect in Bacillus thuringiensis (Bt) cotton fields. Hormone 20-hydroxyecdysone (20E) regulates multiple insect development and physiology events. 20E responses are controlled by pathways triggered by phospholipase C (PLC)-associated proteins. However, 20E-modulated genes and related proteins that can be affected by PLC still remain unknown. Here, isobaric tag for relative and absolute quantitation (iTRAQ) and immunoblotting techniques were used to compare differentially expressed proteins (DEPs) in A. lucorum in response to the treatment of 20E and the PLC inhibitor U73122 as well as their combination. A total of 1,624 non-redundant proteins and 97, 248, 266 DEPs were identified in the 20E/control, U73122/control, and 20E + U73122/control groups, respectively. Only 8 DEPs, including pathogenesis-related protein 5-like, cuticle protein 19.8, trans-sialidase, larval cuticle protein A2B-like, cathepsin L1, hemolymph juvenile hormone-binding protein, ATP-dependent RNA helicase p62-like, and myosin-9 isoform X1, were detected in all three groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the DEPs were involved in diverse signaling pathways. The results were validated by immunoblotting, which highlighted the reliability of proteomics analysis. These findings provided novel insights into the function of PLC in 20E signaling pathway in A. lucorum.

Proteomic analysis suggests that monoterpenes in lemongrass disrupt Ca2+ homeostasis in Haemaphysalis longicornis leading to mitochondrial depolarization and cytotoxicity.

Complex mixtures of bioactive ingredients in plant essential oils present complex chemistries which involve different modes of action. An increasing body of scientific reports has recently focused on the acaricidal activities of plant essential oils attributed to their monoterpene components, but information about their underlying molecular mechanism of action is scarce. Here, after the chemical analysis of lemongrass oil, a proteomic analysis of the ovary, salivary gland, and midgut of Haemaphysalis longicornis exposed to Cymbopogon citratus (lemongrass) essential oil was performed via data-independent acquisition mass spectrometry (DIA-MS) technology to further elucidate the molecular mechanisms involved. Pathway analysis reveals the activation of metabolic pathways mediated by oxidoreductases and transferases. Furthermore, the upregulation of various calcium-associated proteins and the upregulation of cytochrome c1, cytochrome c oxidase polypeptide IV, and programmed cell death protein 6-like isoform X1 suggest a cytotoxic mode of action via the formation of reactive oxygen species (ROS), mitochondrial Ca2+ overload, mitochondrial uncoupling, and depolarization, and ATP depletion leading to either apoptotic or necrotic death. Morphological alterations observed after the RNAi of a major detoxification enzyme (glutathione S-transferase) merit further investigation. Hence, the cytotoxic mode of action exhibited by C. citratus oil could be vital for the development of eco-friendly acaricide.

Comparative transcriptomics analyses and revealing candidate networks and genes involved in lordosis of the Yunlong grouper (Epinephelus moara ♀ × Epinephelus lanceolatus ♂)

Grouper is an economically important fish in China. However, it exhibits a high frequency of skeletal abnormalities, particularly vertebral deformities. The molecular mechanisms underlying fish vertebral deformities are still poorly understood. In this study, a HiSeq™ 4000 platform (Illumina) was used to analyze the transcriptomic profiles of the brain, pituitary, and vertebrae from normal fish (NF) and fish with lordosis (LF) of Yunlong grouper. A total of 87,888 unigenes were assembled with lengths that varied from 201 to 28,922 bp and a N50 length of 2670 bp. A total of 36,268 unigenes were functionally annotated by BLAST alignments. A total of 2875 significantly differentially expressed genes (DEGs) were identified between the NF group and the LF group, including 706 upregulated unigenes and 2169 downregulated unigenes in LF. GO and KEGG pathway enrichment analyses showed that DNA binding, transmembrane receptor activity, cytokine receptor interaction, neuroactive ligand-receptor interaction, calcium signaling pathway and ECM-receptor interaction HIF-1 signaling pathway, and mineral absorption may be involved in the formation of vertebral deformities. Furthermore, weighted gene co-expression network analyses, including three modules (turquoise, yellow, and blue), significantly positively corrected with vertebral deformities. A network map that included these three modules enabled the identification of a series of hub genes, including claudin-22-like (cldn22), fibronectin type III domain-containing protein 1 isoform X2 (fndc1l2), E3 ubiquitin-protein ligase NRDP1-like (rnf41), and Catenin alpha-2 (ctnna1). We found that the levels of most genes in the blue module were closely related to the expression of parvalbumin, thymic CPV3-like isoform X2 (ocm), platelet glycoprotein Ib alpha chain (gp1ba), and matrix metalloproteinase-9 (mmp9), suggesting that this module is associated with skeletal development. Some uncharacterized genes associated with known bone-related genes, including Unigene0067643, Unigene0056862, and Unigene0059867, were detected by a weighted gene co-expression network analysis. A detailed functional investigation of these networks and genes will further improve our understanding of the molecular mechanisms that underlie the formation of lordosis in fish.

Weighted gene coexpression correlation network analysis reveals a potential molecular regulatory mechanism of anthocyanin accumulation under different storage temperatures in \u2018Friar\u2019 plum

BackgroundFlesh is prone to accumulate more anthocyanin in postharvest ‘Friar’ plum (Prunus salicina Lindl.) fruit stored at an intermediate temperature. However, little is known about the molecular mechanism of anthocyanin accumulation regulated by storage temperature in postharvest plum fruit.ResultsTo reveal the potential molecular regulation mechanism of anthocyanin accumulation in postharvest ‘Friar’ plum fruit stored at different temperatures (0 °C, 10 °C and 25 °C), the fruit quality, metabolite profile and transcriptome of its flesh were investigated. Compared to the plum fruit stored at 0 °C and 25 °C, the fruit stored at 10 °C showed lower fruit firmness after 14 days and reduced the soluble solids content after 21 days of storage. The metabolite analysis indicated that the fruit stored at 10 °C had higher contents of anthocyanins (pelargonidin-3-O-glucoside, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside and quercetin-3-O-rutinose), quercetin and sucrose in the flesh. According to the results of weighted gene coexpression correlation network analysis (WGCNA), the turquoise module was positively correlated with the content of anthocyanin components, and flavanone 3-hydroxylase (F3H) and chalcone synthase (CHS) were considered hub genes. Moreover, MYB family transcription factor APL (APL), MYB10 transcription factor (MYB10), ethylene-responsive transcription factor WIN1 (WIN1), basic leucine zipper 43-like (bZIP43) and transcription factor bHLH111-like isoform X2 (bHLH111) were closely related to these hub genes. Further qRT–PCR analysis verified that these transcription factors were specifically more highly expressed in plum flesh stored at 10 °C, and their expression profiles were significantly positively correlated with the structural genes of anthocyanin synthesis as well as the content of anthocyanin components. In addition, the sucrose biosynthesis-associated gene sucrose synthase (SS) was upregulated at 10 °C, which was also closely related to the anthocyanin content of plum fruit stored at 10 °C.ConclusionsThe present results suggest that the transcription factors APL, MYB10, WIN1, bZIP43 and bHLH111 may participate in the accumulation of anthocyanin in ‘Friar’ plum flesh during intermediate storage temperatures by regulating the expression of anthocyanin biosynthetic structural genes. In addition, the SS gene may play a role in anthocyanin accumulation in plum flesh by regulating sucrose biosynthesis.

Identification of Differentially Expressed Genes and Pathways Involved in Growth and Development of Mesona chinensis Benth Under Red- and Blue-Light Conditions.

Mesona chinensis Benth (MCB) is an important Chinese herbal medicine. The plant factories might be one of the ways to solve the shortage of MCB supply. In this study, the MCB seedlings were treated under the red (R) and blue (B) lights in the plant factory. Results showed that the red light promoted the growth and development of MCB in comparison with the blue light. Under the red-light condition, the biomass, plant height, and root characteristics were significantly higher than those under blue-light condition, while the soil and plant analyzer development (SPAD) under the red-light treatment was significantly lower than that under the blue-light treatment. Red light also significantly promoted the content of soluble sugar and pectin of MCB compared with blue light. Transcriptome analysis showed that a total of 4,165 differentially expressed genes (DEGs) were detected including 2,034 upregulated and 2,131 downregulated. Of these, 1,112 DEGs including 410 upregulated and 702 downregulated genes were associated with 111 pathways. Moreover, a total of 8,723 differentially expressed transcription factors (TFs) were identified in R vs. B, and these TFs were distributed in 56 gene families. Metabonomic results revealed that a total of 184 metabolites and 99 differentially expressed metabolites (DEMs) (42 upregulated and 57 downregulated) were identified in the red- and blue-light treatments. Integrative analysis of transcriptome and metabolome unveiled that a total of 24 pathways included 70 compounds (metabolites) and were associated with 28 unigenes. In particular, these pathways included starch and sucrose metabolism, phenylpropanoid biosynthesis, cysteine and methionine metabolism, glycolysis/gluconeogenesis, and pentose and glucuronate interconversions. The unigenes included asparagine synthetase (AS), thymidine kinase (TK), alpha, alpha-trehalose-phosphate synthase (TPS), phosphatase IMPL1 (IMPL1), dihydroflavonol 4-reductase (D4R), and 4-coumarate-CoA ligase-like 6 (4CL6), bifunctional aspartokinase-homoserine dehydrogenase 1 (thrA), and abscisic acid 8′-hydroxylase 2 isoform X1 (ABA8). It was indicated that these pathways and genes might play important roles in the growth and development of MCB. This study laid a foundation for the future research of MCB.

Effects of dietary trehalose on growth, trehalose content, non-specific immunity, gene expression and desiccation resistance of juvenile red claw crayfish (Cherax quadricarinatus)

This study was performed to investigate the effects of dietary trehalose on growth, muscle composition, non-specific immune responses, gene expression and desiccation resistance of juvenile red claw crayfish (Cherax quadricarinatus). A total of 540 (body weight of 0.41 ± 0.05) crayfish were randomly divided into six groups for a feeding experiment. Six diets with trehalose levels at 0 (Diet 1), 1 (Diet 2), 2 (Diet 3), 5 (Diet 4), 10 (Diet 5) and 15 (Diet 6) g kg−1 were prepared to feed juvenile red claw crayfish for 8 weeks. The results showed that the weight gain rate (WGR) and specific growth rate (SGR) of crayfish in Diet 4, Diet 5 and Diet 6 groups were significantly improved compared with the control group (Diet 1). Muscle crude protein contents of crayfish fed Diet 4, Diet 5 and Diet 6 were significantly higher than those of the control group. The activities of superoxide dismutase (SOD) and alkaline phosphatase (AKP) in hepatopancreas and hemolymph of crayfish for Diet 4, Diet 5, and Diet 6 groups were significantly increased while malondialdehyde (MDA) content was significantly reduced when compared with the control. The total antioxidant capacity (T-AOC), catalase (CAT) and glutathione peroxidase (GPx) activities in the hepatopancreas and hemolymph of crayfish fed Diet 5 and Diet 6 were significantly higher than those in the control group. However, acid phosphatase (ACP) activity was not significantly different among all experimental groups. The hepatopancreas and intestine trehalose contents of crayfish showed an upward trend with the increase of dietary trehalose levels. Compared with the control group, supplementation of 5–15 g kg−1 trehalose in the feed up-regulated the expression levels of GPx, C-type lysozyme (C-LZM), antilipolysacchride factor (ALF), facilitated trehalose transporter homolog isoform X2 (Tret1-2) and facilitated trehalose transporter isoform X4 (Tret1-4) mRNA. In addition, supplementation of 5–15 g kg−1 trehalose in the feed could improve the survival rate of red claw crayfish under desiccation stress. These results suggested that supplementation of 5–15 g kg−1 trehalose in feed could significantly improve the growth performance, muscle protein, non-specific immunity and desiccation resistance of juvenile red claw crayfish.