Genes Encoding Heat Shock Proteins Research Articles

Transcriptome Analysis Reveals Genes and Pathways Associated with Drought Tolerance of Early Stages in Sweet Potato (Ipomoea batatas (L.) Lam.).

The yield of sweet potato [Ipomoea batatas (L.) Lam] can be easily threatened by drought stress. Typically, early stages like the seedling stage and tuber-root expansion stage are more vulnerable to drought stress. In this study, a highly drought-tolerant sweet potato cultivar "WanSu 63" was subjected to drought stress at both the seedling stage (15 days after transplanting, 15 DAT) and the tuber-root expansion stage (45 DAT). Twenty-four cDNA libraries were constructed from leaf segments and root tissues at 15 and 45 DAT for Next-Generation Sequencing. A total of 663, 063, and 218 clean reads were obtained and then aligned to the reference genome with a total mapped ratio greater than 82.73%. A sum of 7119, 8811, 5463, and 930 differentially expressed genes were identified from leaves in 15 days (L15), roots in 15 days (R15), leaves in 45 days (L45), and roots in 45 days (R45), respectively, in drought stress versus control. It was found that genes encoding heat shock proteins, sporamin, LEA protein dehydrin, ABA signaling pathway protein gene NCED1, as well as a group of receptor-like protein kinases genes were enriched in differentially expressed genes. ABA content was significantly higher in drought-treated tissues than in the control. The sweet potato biomass declined sharply to nearly one-quarter after drought stress. In conclusion, this study is the first to identify the differentially expressed drought-responsive genes and signaling pathways in the leaves and roots of sweet potato at the seedling and root expansion stages. The results provide potential resources for drought resistance breeding of sweet potato.

Upgraded durian genome reveals the role of chromosome reshuffling during ancestral karyotype evolution, lignin biosynthesis regulation, and stress tolerance.

Durian (Durio zibethinus) is a tropical fruit that has a unique flavor and aroma. It occupies a significant phylogenetic position within the Malvaceae family. Extant core-eudicot plants are reported to share seven ancestral karyotypes that have undergone reshuffling, resulting in an abundant genomic diversity. However, the ancestral karyotypes of the Malvaceae family, as well as the evolution trajectory leading to the 28 chromosomes in durian, remain poorly understood. Here, we report the high-quality assembly of the durian genome with comprehensive comparative genomic analyses. By analyzing the collinear blocks between cacao and durian, we inferred 11 Malvaceae ancestral karyotypes. These blocks were present in a single-copy form in cacao and mainly in triplicates in durian, possibly resulting from a recent whole genome triplication (WGT) event that led to hexaploidization of the durian genome around 20 (17-24) million years ago. A large proportion of the duplicated genes in durian, such as those involved in the lignin biosynthesis module for phenylpropane biosynthesis, are derived directly from whole genome duplication, which makes it an important force in reshaping its genomic architecture. Transcriptome studies have revealed that genes involved in feruloyl-CoA formations were highly preferentially expressed in fruit peels, indicating that the thorns produced on durian fruit may comprise guaiacyl and syringyl lignins. Among all the analyzed transcription factors (TFs), members of the heat shock factor family (HSF) were the most significantly upregulated under heat stress. All subfamilies of genes encoding heat shock proteins (HSPs) in the durian genome appear to have undergone expansion. The potential interactions between HSF Dzi05.397 and HSPs were examined and experimentally verified. Our study provides a high-quality durian genome and reveals the reshuffling mechanism of ancestral Malvaceae chromosomes to produce the durian genome. We also provide insights into the mechanism underlying lignin biosynthesis and heat stress tolerance.

Trehalose 6-phosphate synthase gene rdtps1 contributes to thermal acclimation in Rhyzopertha dominica

BackgroundThe lesser grain borer (Rhyzopertha dominica), a worldwide primary pest of stored grain, causes serious economic losses and threatens stored food safety. R. dominica can respond to changes in temperature, especially the adaptability to heat. In this study, transcriptome analysis of R. dominica exposed to different temperatures was performed to elucidate differences in gene expression and the underling molecular mechanism.ResultsIsoform-sequencing generated 17,721,200 raw reads and yielded 20,416 full-length transcripts. A total of 18,880 (92.48%) transcripts were annotated. We extracted RNA from R. dominica reared at 5 °C (cold stress), 15 °C (cold stress), 27 °C (ambient temperature) and 40 °C (heat stress) for RNA-seq. Compared to those of control insects reared at 27 °C, 119, 342, and 875 differentially expressed genes (DEGs) were identified at 5 °C, 15 °C, and 40 °C, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that pathways associated with “fatty acid metabolism”, “fatty acid biosynthesis”, “AMPK signaling pathway”, “neuroactive ligand receptor interaction”, and “longevity regulating pathway-multiple species” were significantly enriched. The functional annotation revealed that the genes encoding heat shock proteins (HSPs), fatty acid synthase (FAS), phospholipases (PLA), trehalose transporter (TPST), trehalose 6-phosphate synthase (TPS), and vitellogenin (Vg) were most likely involved in temperature regulation, which was also validated by RT-qPCR. Seven candidate genes (rdhsp1, rdfas1, rdpla1, rdtpst1, rdtps1, rdvg1, and rdP450) were silenced in the RNA interference (RNAi) assay. RNAi of each candidate gene suggested that inhibiting rdtps1 expression significantly decreased the trehalose level and survival rate of R. dominica at 40 °C.ConclusionsThese results indicated that trehalose contributes to the high temperature resistance of R. dominica. Our study elucidates the molecular mechanisms underlying heat tolerance and provides a potential target for the pest management in R. dominica.

The development of abamectin resistance in Liriomyza trifolii and its contribution to thermotolerance.

Liriomyza trifolii is an economically significant, invasive pest of horticultural and vegetable crops. The larvae form tunnels in foliage and hasten senescence and death. Outbreaks of L. trifolii often erupt in hot weather and are driven by thermotolerance; furthermore, the poor effectiveness of pesticides has made outbreaks more severe. But it is still unclear whether the development of insecticide tolerance will contribute to thermotolerance in L. trifolii. To explore potential synergistic relationships between insecticide exposure and thermotolerance in L. trifolii, we first generated an abamectin-resistant (AB-R) strain. Knockdown behavior, eclosion and survival rates, and expression levels of genes encoding heat shock proteins (Hsps) in L. trifolii were then examined in AB-R and abamectin-susceptible (AB-S) strains. Our results demonstrated that long-term selection pressure for abamectin resistance made L. trifolii more prone to develop cross-resistance to other insecticides containing similar ingredients. Furthermore, the AB-R strain exhibited enhanced thermotolerance and possessed an elevated critical thermal maximum temperature, and upregulated expression levels of Hsps during heat stress. Collectively, our results indicate that thermal adaptation in L. trifolii was accompanied by emerging abamectin resistance. This study provides a theoretical basis for investigating the synergistic or cross-adaptive mechanisms that insects use to cope with adversity and demonstrates the complexity of insect adaptation to environmental and chemical stress. © 2023 Society of Chemical Industry.

Alleviation of heat stress in electrostimulation assisted SBR treating thermophilic phenol wastewater: Performance, microbial community assembly, heat resistance mechanism and antibiotic resistance genes

An electrostimulation assisted sequencing batch reactor (ESBR) has excellent performance in improving phenol treatment performance and sludge characteristics under high temperature. The sludge volume index (SVI) and extracellular polymeric substances (EPS) content of ESBR were significantly reduced by 61.7% and 74.1% compared to SBR, respectively. The content of EPS was positively correlated with SVI, indicating that electrostimulation enhanced the settleability of sludge. Microbial community assembly under electrostimulation could be dominated by the deterministic process. The genes encoding heat shock proteins (Hsp15 and Hsp33) and synthesis genes encoding trehalose (TPS1 and otsA), and glycogen (GSY1) were up-regulated. The activation of the antioxidant system in ESBR inhibited the production of ROS, thereby hindering the transmission of ARGs under high temperature. The coexistence pattern of ARGs and genera indicated that the genus Ralstonia, Rhizobium, Schaalia, and Streptococcus were speculated to be the main hosts, thus electrostimulation reduced the abundance of ARGs hosts, thereby reducing the dissemination of ARGs under high temperature.

Исследование теплового воздействия фемтосекундных лазерных импульсов на эмбрионы мыши в рамках процедуры вспомогательного хетчинга

Impaired hatching of the embryo from the zona pellucida (ZP), the specialized protective shell, immediately before implantation is one of the factors of infertility. Hatching impairment is often due to the ZP hardening or thickening. In such cases, the laser assisted hatching procedure is used to overcome infertility. During this procedure a hole is drilled in the ZP facilitating the embryo release. The question of the safe use of laser for assisted hatching remains open, since laser beam can heat the environment and cause thermal shock in embryos. The study was aimed to assess safety of the mouse embryo femtosecond laser exposure during the assisted hatching procedure using the embryo viability and HSP gene expression assessment methods. A new type of pulsed laser was used in the study for the ZP dissection — the femtosecond laser. The energy of such pulses was two orders of magnitude lower than the energy of laser dissectors currently used in the clinics. To assess the femtosecond laser exposure to the embryo, the house mouse (Mus musculus) embryos were stained with fluorescence dyes, and expression of the genes encoding heat shock proteins (Hsp90aa1 and Hspa5) was assessed. The embryonic cells remained viable after the laser assisted hatching procedure involving the use of a femtosecond laser, while expression levels of the genes encoding heat shock proteins were slightly increased compared to the negative control group (p = 0.408).

Astrocyte transcriptomic changes along the spatiotemporal progression of Alzheimer’s disease

AbstractBackgroundAlzheimer’s disease (AD) is defined by widespread accumulation of Ab plaques and phospho‐tau neurofibrillary tangles throughout the brain, with the latter following a stereotypical hierarchical spatiotemporal pattern along neural networks. These AD neuropathological changes are accompanied by a dramatic loss of synapses and neurons as well as prominent morphological and functional changes of astrocytes, collectively termed reactive astrogliosis. Astrocytes also play a critical role in maintaining brain homeostasis. However, their changes along the spatiotemporal progression of AD neuropathology remain largely unknown. To investigate these astrocyte changes, we performed single‐nucleus RNA‐sequencing on 5 brain regions along the stereotypical progression of AD neuropathology from 32 donors representing the entire normal aging‐severe AD spectrum.MethodNuclei were isolated from five brain areas of n = 32 donors with increasing AD neuropathology (total n = 160 samples). The five brain areas were chosen based on their hierarchical accumulation of tau pathology: entorhinal cortex (EC)> inferior temporal gyrus (BA20) > dorsolateral prefrontal cortex (BA46) > secondary visual cortex (V2 or BA18/19) > primary visual cortex (V1 or BA17). To enrich in astrocytic nuclei, NEUN+ and OLIG2+ nuclei were separated via FACS, whereas NEUN‐/OLIG2‐ nuclei (including astrocytes) were subjected to snRNA‐seq resulting in a transcriptomic dataset of 629,755 astrocyte nuclei, which were subsequently examined for common and region‐specific AD related changes.ResultWe discovered temporal gene‐expression‐trajectories with gene sets differentially activated at various disease stages. Surprisingly, a gene set enriched in proteostasis (genes encoding heat shock proteins) and energy metabolism (e.g., ATP synthases) was upregulated at late‐stage but unexpectedly returned to baseline levels at end‐stage, suggesting exhaustion of response in “burnt‐out” astrocytes. The spatial gene‐expression‐trajectories revealed that genes related to tripartite synapses (e.g., GLUL, GRIA2, NRXN1, SLC1A2) are dysregulated in parallel to the stereotypical progression of tangle pathology across regions. In addition, we identified substantial astrocyte heterogeneity across brain regions with a continuum from homeostatic to reactive cells through “intermediate” transitional states.ConclusionOur astrocyte snRNA‐seq dataset encompassing five regions of control and AD brains revealed distinct spatial and temporal gene trajectories, suggesting complex astrocytic dysfunction with AD progression.

The miR-221/222 cluster regulates hematopoietic stem cell quiescence and multipotency by suppressing both Fos/AP-1/IEG pathway activation and stress-like differentiation to granulocytes.

Throughout life, hematopoietic stem cells (HSCs), residing in bone marrow (BM), continuously regenerate erythroid/megakaryocytic, myeloid, and lymphoid cell lineages. This steady-state hematopoiesis from HSC and multipotent progenitors (MPPs) in BM can be perturbed by stress. The molecular controls of how stress can impact hematopoietic output remain poorly understood. MicroRNAs (miRNAs) as posttranscriptional regulators of gene expression have been found to control various functions in hematopoiesis. We find that the miR-221/222 cluster, which is expressed in HSC and in MPPs differentiating from them, perturbs steady-state hematopoiesis in ways comparable to stress. We compare pool sizes and single-cell transcriptomes of HSC and MPPs in unperturbed or stress-perturbed, miR-221/222-proficient or miR-221/222-deficient states. MiR-221/222 deficiency in hematopoietic cells was induced in C57BL/6J mice by conditional vav-cre-mediated deletion of the floxed miR-221/222 gene cluster. Social stress as well as miR-221/222 deficiency, alone or in combination, reduced HSC pools 3-fold and increased MPPs 1.5-fold. It also enhanced granulopoisis in the spleen. Furthermore, combined stress and miR-221/222 deficiency increased the erythroid/myeloid/granulocytic precursor pools in BM. Differential expression analyses of single-cell RNAseq transcriptomes of unperturbed and stressed, proficient HSC and MPPs detected more than 80 genes, selectively up-regulated in stressed cells, among them immediate early genes (IEGs). The same differential single-cell transcriptome analyses of unperturbed, miR-221/222-proficient with deficient HSC and MPPs identified Fos, Jun, JunB, Klf6, Nr4a1, Ier2, Zfp36-all IEGs-as well as CD74 and Ly6a as potential miRNA targets. Three of them, Klf6, Nr4a1, and Zfp36, have previously been found to influence myelogranulopoiesis. Together with increased levels of Jun, Fos forms increased amounts of the heterodimeric activator protein-1 (AP-1), which is known to control the expression of the selectively up-regulated expression of the IEGs. The comparisons of single-cell mRNA-deep sequencing analyses of socially stressed with miR-221/222-deficient HSC identify 5 of the 7 Fos/AP-1-controlled IEGs, Ier2, Jun, Junb, Klf6, and Zfp36, as common activators of HSC from quiescence. Combined with stress, miR-221/222 deficiency enhanced the Fos/AP-1/IEG pathway, extended it to MPPs, and increased the number of granulocyte precursors in BM, inducing selective up-regulation of genes encoding heat shock proteins Hspa5 and Hspa8, tubulin-cytoskeleton-organizing proteins Tuba1b, Tubb 4b and 5, and chromatin remodeling proteins H3f3b, H2afx, H2afz, and Hmgb2. Up-regulated in HSC, MPP1, and/or MPP2, they appear as potential regulators of stress-induced, miR-221/222-dependent increased granulocyte differentiation. Finally, stress by serial transplantations of miR-221/222-deficient HSC selectively exhausted their lymphoid differentiation capacities, while retaining their ability to home to BM and to differentiate to granulocytes. Thus, miR-221/222 maintains HSC quiescence and multipotency by suppressing Fos/AP-1/IEG-mediated activation and by suppressing enhanced stress-like differentiation to granulocytes. Since miR-221/222 is also expressed in human HSC, controlled induction of miR-221/222 in HSC should improve BM transplantations.

LAMP‐based detection of Monilinia fructigena, Monilinia polystroma and Monilinia fructicola in latently infected apple fruit

AbstractBrown rot in apples is caused by fungi belonging to the genus Monilinia and occurs annually in orchards with varying severity. The symptoms of the disease on apple fruits may occur during harvest and can also develop much later in storage. Therefore, from a practical point of view, it is very important to be able to test a sample of asymptomatic apples before putting them in storage or sending them to distant markets. Thus, there is a need to develop a quick and sensitive method of detecting disease causal agents in asymptomatically infected fruit. For this purpose, a procedure for DNA isolation from the skin of apples was developed, and a set of primers for the loop‐mediated isothermal amplification (LAMP) reaction was designed based on a fragment of the hsp60 gene encoding heat shock protein 60 of Monilinia fungi that are pathogenic to apple: M. fructigena, M. fructicola and M. polystroma. The method was validated by reactions with DNA from infected apple fruit exhibiting brown rot symptoms and DNA from infected apple fruit without visible brown rot symptoms (latent infection). To increase the sensitivity of detection before the LAMP reaction, preamplification of the target DNA was carried out. The sensitivity of detection in the LAMP assay for the DNA of all three tested species of fungi was 2 pg. The DNA preamplification significantly increased the sensitivity of Monilinia DNA detection to 0.1 pg. This study shows that the developed LAMP method can be used to detect Monilinia fungi in asymptomatically infected apples.

Characterization of genes encoding heat shock proteins reveals a differential response to temperature in two geographic populations of Liriomyza trifolii (Diptera: Agromyzidae)

Liriomyza trifolii is a significant, invasive pest that damages horticultural crops and vegetables. The distribution of L. trifolii is influenced by temperature, and prior research has demonstrated that variations in thermal adaptability differ among geographic populations of the insect. Heat shock proteins (Hsps) are involved in adaptation to temperatures; however, the underlying molecular mechanism for thermal adaption in different L. trifolii populations remains unclear. This study examines the temperature adaptability of two L. trifolii populations from Hainan (HN) and Jiangsu (JS) provinces. The results indicate that the HN population has a higher survival rate and a higher critical thermal maximum (CTmax) than the JS population under high temperature stress. Transcriptome data at 42 °C revealed that the JS population has more differentially expressed genes (DEGs) than the HN population, while the HN population has more upregulated DEGs. The two populations were similar in functional annotation of DEGs, and a large number of Hsps were upregulated. However, the HN population had larger numbers and higher expression levels of Hsps during heat stress as compared to the JS population. Additionally, the expression patterns of differentially expressed Hsps varied between the HN and JS populations in response to different elevated temperatures. Notably, the transcription levels of Hsp70s were higher in the HN population as compared to the JS population, while the expression level of genes encoding small heat shock proteins was higher in the JS population. These findings have significant scientific value in understanding the underlying mechanism of temperature adaption in L. trifolii and provide a fresh perspective on the distribution of this invasive pest.

Involvement of RpoN in regulating stress response, biofilm formation and virulence of Vibrio mimicus

RpoN, an alternative sigma factor, plays a crucial role in response to changes in environmental cues and pathogenicity. Vibrio mimicus, a serious pathogen in both fresh and seawater aquaculture industries, has garnered considerable attention. Nevertheless, reports of rpoN in V. mimicus remain extremely scarce. In this study, we constructed the rpoN-RNAi strain to investigate the role of rpoN in V. mimicus using phenotypic and RNA-seq analysis. The data indicated that rpoN positively regulated the resistance of V. mimicus to H2O2, ethanol and acid, while negatively regulating the resistance to alkali and heat, and had no influence on the resistance to NaCl. Over the period of 12 to 72 h, the biofilm formation ability of rpoN-RNAi strain was decreased by 0.29 to 0.84-fold compared to wild-type strain. Compared to wild strain, the siderophore production was decreased from 85.9% to 60.3% in rpoN-RNAi strain. Furthermore, the LD50 increased from 6.16 × 105 to 3.92 × 106 CFU/mL in Macrobrachium nipponense through artificial infection test after rpoN silencing. Transcriptome analysis revealed that several well-known virulence-related genes were down-regulated in rpoN-RNAi strain. These genes included the type VI secretion system genes (vipB, vipA, impI, and vasA), type IV pilus assembly protein (flpB-G), outer membrane proteins (ompU and ompA), and hemolysin. Additionally, stress-protective genes encoding heat shock protein, oxidoreductase, aldehyde dehydrogenase, quorum sensing system, etc., were also down-regulated by rpoN-RNAi. Transcriptome analysis was consistent with the observed phenotypes in rpoN-RNAi strain. These results revealed that rpoN is a critical determinant for bacterial fitness and pathogenicity, laying the groundwork for further characterization of RpoN regulatory networks.

Transcriptome profiling brings new insights into the ethanol stress responses of Spathaspora passalidarum.

Spathaspora passalidarum is a xylose-fermenting microorganism promising for the fermentation of lignocellulosic hydrolysates. This yeast is more sensitive to ethanol than Saccharomyces cerevisiae for unclear reasons. An RNA-seq experiment was performed to identify transcriptional changes in S. passalidarum in response to ethanol and gain insights into this phenotype. The results showed the upregulation of genes associated with translation and the downregulation of genes encoding proteins involved in lipid metabolism, transporters, and enzymes from glycolysis and fermentation pathways. Our results also revealed that genes encoding heat-shock proteins and involved in antioxidant response were upregulated, whereas the osmotic stress response of S. passalidarum appears impaired under ethanol stress. A pseudohyphal morphology of S. passalidarum colonies was observed in response to ethanol stress, which suggests that ethanol induces a misperception of nitrogen availability in the environment. Changes in the yeast fatty acid profile were observed only after 12 h of ethanol exposure, coinciding with the recovery of the yeast xylose consumption ability. These findings suggest that the lack of fast membrane lipid adjustments, the halt in nutrient absorption and cellular metabolism, and the failure to induce the expression of osmotic stress-responsive genes are the main aspects underlying the low ethanol tolerance of S. passalidarum. KEY POINTS: • Ethanol stress halts Spathaspora passalidarum metabolism and fermentation • Genes encoding nutrient transporters showed downregulation under ethanol stress • Ethanol induces a pseudohyphal cell shape, suggesting a misperception of nutrients.

Role of the Structural and Functional Genes Encoding Heat Shock Proteins in Biosurfactant Synthesis by Rhodococcus pyridinivorans 5Ap

Role of the Structural and Functional Genes Encoding Heat Shock Proteins in Biosurfactant Synthesis by Rhodococcus pyridinivorans 5Ap

Histological and transcriptomic analysis of muscular atrophy associated with depleted flesh pigmentation in Atlantic salmon (Salmo salar) exposed to elevated seawater temperatures

Tasmania is experiencing increasing seawater temperatures during the summer period which often leads to thermal stress-induced starvation events in farmed Atlantic salmon, with consequent flesh pigment depletion. Our previous transcriptomic studies found a link between flesh pigmentation and the expression of genes regulating lipid metabolism accompanied by feeding behavior in the hindgut. However, the impact of prolonged exposure to elevated water temperature on muscle structural integrity and molecular mechanisms in muscle underlying pigment variation has not been elucidated to date. In this study, we investigated the effect of prolonged exposure to elevated water temperature on the farmed salmon flesh pigmentation and structural integrity, using muscle histological and transcriptomic analysis. On April 2019, after the end of the summer, two muscle regions of the fish fillet, front dorsal and back central (usually the most and least affected by depletion, respectively), were sampled from fifteen fish (weighing approximately 2 kg and belonging to the same commercial population split in two cages). The fish represented three flesh color intensity groups (n = 5 fish per group) categorized according to general level of pigmentation and presence of banding (i.e. difference in color between the two regions of interest) as follows: high red color-no banding (HN), high red color-banded (HB) and Pale fish. Histological analysis showed a distinction between the flesh color intensity phenotypes in both muscle regions. Muscle fibers in the HB fish were partly degraded, while they were atrophied and smaller in size in Pale fish compared to HN fish. In the Pale fish, interstitial spaces between muscle fibers were also enlarged. Transcriptomic analysis showed that in the front dorsal region of the HN fish, genes encoding collagens, calcium ion binding and metabolic processes were upregulated while genes related to lipid and fatty acid metabolism were downregulated when compared to HB fish. When comparing the back central region of the three phenotypes, actin alpha skeletal muscle and myosin genes were upregulated in the HN and HB fish, while tropomyosin genes were upregulated in the Pale fish. Also, genes encoding heat shock proteins were upregulated in the HN fish, while genes involving lipid metabolism and proteolysis were upregulated in the Pale fish. Starvation, likely caused by thermal stress during prolonged periods of elevated summer water temperatures, negatively affects energy metabolism to different extents, leading to localized or almost complete flesh color depletion in farmed Atlantic salmon. Based on our results, we conclude that thermal stress is responsible not only for flesh discoloration but also for loss of muscle integrity, which likely plays a key role in pigment depletion.

Graphene Oxides (GOs) with Different Lateral Dimensions and Thicknesses Affect the Molecular Response in Chironomus riparius.

Graphene oxide (GO) materials possess physicochemical properties that facilitate their application in the industrial and medical sectors. The use of graphene may pose a threat to biota, especially aquatic life. In addition, the properties of nanomaterials can differentially affect cell and molecular responses. Therefore, it is essential to study and define the possible genotoxicity of GO materials to aquatic organisms and their ecosystems. In this study, we investigated the changes in the expression of 11 genes in the aquatic organism Chironomus riparius after 96 h of exposure to small GOs (sGO), large GOs (lGO) and monolayer GOs (mlGO) at 50, 500 and 3000 μg/L. Results showed that the different genes encoding heat shock proteins (hsp90, hsp70 and hsp27) were overexpressed after exposure to these nanomaterials. In addition, ATM and NLK-the genes involved in DNA repair mechanisms-were altered at the transcriptional level. DECAY, an apoptotic caspase, was only activated by larger size GO materials, mlGO and lGO. Finally, the gene encoding manganese superoxide dismutase (MnSOD) showed higher expression in the mlG O-treated larvae. The lGO and mlGO treatments indicated high mRNA levels of a developmental gene (FKBP39) and an endocrine pathway-related gene (DRONC). These two genes were only activated by the larger GO materials. The results indicate that larger and thicker GO nanomaterials alter the transcription of genes involved in cellular stress, oxidative stress, DNA damage, apoptosis, endocrine and development in C. riparius. This shows that various cellular processes are modified and affected, providing some of the first evidence for the action mechanisms of GOs in invertebrates. In short, the alterations produced by graphene materials should be further studied to evaluate their effect on the biota to show a more realistic scenario of what is happening at the molecular level.

Rice DST transcription factor negatively regulates heat tolerance through ROS-mediated stomatal movement and heat-responsive gene expression.

Plants are frequently subjected to a broad spectrum of abiotic stresses including drought, salinity and extreme temperatures and have evolved both common and stress-specific responses to promote fitness and survival. Understanding the components and mechanisms that underlie both common and stress-specific responses can enable development of crop plants tolerant to different stresses. Here, we report a rice heat stress-tolerant 1 (hst1) mutant with increased heat tolerance. HST1 encodes the DST transcription factor, which also regulates drought and salinity tolerance. Increased heat tolerance of hst1 was associated with suppressed expression of reactive oxygen species (ROS)-scavenging peroxidases and increased ROS levels, which reduced water loss by decreasing stomatal aperture under heat stress. In addition, increased ROS levels enhanced expression of genes encoding heat shock protein (HSPs) including HSP80, HSP74, HSP58 and small HSPs. HSPs promote stabilization of proteins and protein refolding under heat stress and accordingly mutation of HST1 also improved reproductive traits including pollen viability and seed setting under high temperature. These results broaden the negative roles of DST in abiotic stress tolerance and provide important new insights into DST-regulated tolerance to diverse abiotic stresses through both shared and stress-specific mechanisms.

Comparative transcriptome analysis of Callosobruchus chinensis (L.) (Coleoptera: Chrysomelidae-Bruchinae) after heat and cold stress exposure

Callosobruchus chinensis is regarded as one of the cosmopolitan pests of legume crops and can cause tremendous losses to a variety of beans. This study focused on comparative transcriptome analyses of C. chinensis exposed to 45 °C (heat stress), 27 °C (ambient temperature) and −3 °C (cold stress) for 3 h to investigate the gene differences and underlying molecular mechanisms. There were 402 and 111 differentially expressed genes (DEGs) identified in the heat and cold stress treatments, respectively. “cell process”, “cell” and “binding” were the main enriched functions and biological processes revealed by gene ontology (GO) analysis. The clusters of orthologous genes (COG) showed that DEGs were assigned to the categories: “posttranslational modification, protein turnover, chaperones”, “lipid transport and metabolism”, and “general function prediction only”. With respect to the Kyoto Encyclopedia of Genes and Genomes (KEGG), the “longevity regulating pathway-multiple species”, “carbon metabolism”, “peroxisome”, “protein processing in endoplasmic”, “glyoxylate and dicarboxylate metabolism” pathways were significantly enriched. The annotation and enrichment analysis revealed that genes encoding heat shock proteins (Hsps) and cuticular proteins were significantly upregulated under high and low-temperature stresses, respectively. In addition, some DEGs encoding “Protein lethal essential for life”, “Reverse transcriptase”, “DnaJ domain”, “Cytochrome” and “Zinc finger protein” were also upregulated to varying degrees. Transcriptomic data were validated using qRT‒PCR, which confirmed that they were consistent. In this paper, the temperature tolerance of C. chinensis adults was evaluated and the results showed that female adults were more sensitive to heat and cold stress than males, and the upregulation of heat shock protein and epidermal protein was the largest in DEGs after heat and cold stress, respectively. These findings provide a reference for further understanding the biological characteristics of C. chinensis adults and the molecular mechanisms underlying the response to low and high temperatures.

RNA-sequencing Reveals Differentially Expressed Genes of Laying Hens Fed Baihu Decoction Under Heat Shock.

Egg production, an important economic trait in the poultry industry, is sensitive to heat stress. The hypothalamus is a crucial center for thermoregulation by detecting temperature changes and regulating the autonomic nervous system in poultry. Baihu decoction (BH), which contains four ingredients (Rhizoma Anemarrhenae, Gypsum Fibrosum, Radix Glycyrrhizae, and Semen Oryzae Nonglutinosae), is a traditional Chinese medicinal formula for clearing heat. Our study aimed to investigate the changes in gene transcription levels in the hypothalamus of laying hens treated with heat stress with and without BH using RNA sequencing. A total of 223 differentially expressed genes (DEGs) were identified in the heat-treated group compared with the control group and 613 DEGs were identified in the BH group compared with the heat-treated group. Heat shock led to significant changes in the expression of multiple genes involved in the "neuroactive ligand-receptor interaction" pathway. Moreover, feeding BH led to significant upregulation in the expression of eight genes encoding heat shock proteins (HSPs), which were highlighted as candidates to control the "protein processing in the endoplasmic reticulum (ER)" pathway. These results provide the novel insight that BH responds to heat stress by participating in regulation of the ER signaling pathway and HSPs expression.

Mechanistic effects of microwave radiation on pupal emergence in the leafminer fly, Liriomyza trifolii.

Liriomyza trifolii is a significant pest of vegetable and ornamental crops across the globe. Microwave radiation has been used for controlling pests in stored products; however, there are few reports on the use of microwaves for eradicating agricultural pests such as L. trifolii, and its effects on pests at the molecular level is unclear. In this study, we show that microwave radiation inhibited the emergence of L. trifolii pupae. Transcriptomic studies of L. trifolii indicated significant enrichment of differentially expressed genes (DEGs) in 'post-translational modification, protein turnover, chaperones', 'sensory perception of pain/transcription repressor complex/zinc ion binding' and 'insulin signaling pathway' when analyzed with the Clusters of Orthologous Groups, Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes databases, respectively. The top DEGs were related to reproduction, immunity and development and were significantly expressed after microwave radiation. Interestingly, there was no significant difference in the expression of genes encoding heat shock proteins or antioxidant enzymes in L. trifolii treated with microwave radiation as compared to the untreated control. The expression of DEGs encoding cuticular protein and protein takeout were silenced by RNA interference, and the results showed that knockdown of these two DEGs reduced the survival of L. trifolii exposed to microwave radiation. The results of this study help elucidate the molecular response of L. trifolii exposed to microwave radiation and provide novel ideas for control.

Molecular effects of polystyrene nanoplastics toxicity in zebrafish embryos (Daniorerio)

Plastics pose a health hazard to living beings and the environment. Plastic degradation produces nano-sized plastic particles (NPs) that end up in terrestrial and aquatic ecosystems, including oceans, rivers, and lakes. Their presence in air, drinking water, sediments, food, and personal care products leads to a variety of exposure routes for living beings, including humans. The toxicity mechanisms of these nanomaterials (NMs) in living organisms and ecosystems are currently unknown, making it a priority to understand their effects at the molecular and cellular levels. The zebrafish (Zf) (Danio rerio) is a model organism which has a high homology with humans and has been widely used to assess the hazard of different xenobiotics. In this study, the expression changes of different genes in 120 hpf Zf embryos (Zfe) after exposure to polystyrene (PS) NPs (30 nm) at concentrations of 0.1, 0.5 and 3 ppm were investigated. The results showed that the gene encoding heat shock protein (hsp70) was down-regulated in a dose-dependent manner. The genes encoding superoxide dismutase (SOD 1 and SOD 2), apoptotic genes (cas 1 and cas 8) and interleukin 1-β (il1β) were activated at the concentration of 3 ppm PS NP, while the anti-apoptotic gene Bcl2α was inhibited at 0.5 and 3 ppm. In addition, the neurotransmitter-related gene Acetyl-Cholinesterase (ache) was significantly inhibited and the DNA repair genes (gadd45α and rad51) were also down-regulated. In contrast, the mitochondrial metabolism-related gene cox1 did not alter its expression in any of the treatments. Most of the changes in gene expression occurred at the highest concentration of NPs. Overall, the results indicated that NPs generated cellular stress that caused certain alterations in normal gene expression (oxidative stress, apoptotic and inflammatory processes, neurotoxicity and anti-apoptotic proteins), but did not cause any mortality after 120 hpf exposure at the three concentrations assayed. These results highlight the need for further studies investigating the effects, at the molecular level, of these materials in humans and other living organisms.