Heat Shock Protein Genes Research Articles

Safe concentration, unsafe effects: Impact of BPA on antioxidant function in the hepatopancreas and ovarian gene expression in oriental river prawns (Macrobrachium nipponense)

This study investigated the effects of Bisphenol A (BPA), a common endocrine-disrupting chemical, on the antioxidant enzyme activities in the hepatopancreas and the expression of genes related to ovarian development in oriental river prawns (Macrobrachium nipponense). The 24hLC50 and 48hLC50 values for BPA were 80.59 mg/L and 63.90 mg/L, respectively, with a safe concentration of 12.06 mg/L. Prawns were exposed to low (4.85 mg/L), safe (12.06 mg/L), and high (30.00 mg/L) concentrations of BPA for 10 days to measure enzyme activities, and for 20 days followed by 7 days in BPA-free water to measure gene expression.Short-term exposure (12 h, 1d, 3d) to low concentration BPA did not significantly affect superoxide dismutase (SOD) activity in the hepatopancreas (P > 0.05), but long-term exposure (6d, 10d) significantly reduced SOD activity (P < 0.05). Catalase (CAT) activity showed no significant changes throughout the low concentration exposure period (P > 0.05). At safe and high concentrations, SOD and CAT activities significantly decreased after 12 h of exposure (P < 0.05).BPA affected heat shock protein 90 (HSP90) expression in the ovary, with low concentration BPA significantly upregulating HSP90 after 1 day (P < 0.05), but returning to normal levels after 10 and 20 days. At the safe concentration, HSP90 was significantly upregulated at all three sampling points (1d, 10d, 20d) (P < 0.05), while high concentration exposure led to significant upregulation only on day 10 (P < 0.05).Low concentration BPA had no significant effect on Cathepsin B (CB) and Cathepsin L (CL) gene expression in the ovaries (P > 0.05). However, safe concentration exposure promoted CB expression on days 1, 10, and 20 (P < 0.05), while high concentration exposure significantly increased CB expression on day 1 (P < 0.05), with levels returning to normal on days 10 and 20. CL expression significantly increased after 20 days of exposure to both safe and high concentrations (P < 0.05). Gene expression levels in the ovaries returned to normal after transfer to BPA-free water, with HSP90 and CB normalizing by day 1, and CL by day 7.These results indicate that even safe concentrations of BPA impose stress on the hepatopancreas and increase the expression of HSP90, CB, and CL genes in the ovaries, affecting ovarian development. And, these effects are reversible within a certain period after the removal of BPA.

Toxic effects of ZnO NPs on immune response and tissue pathology in Mytilus galloprovincialis

Nano-zinc oxide (ZnO NPs), as widely used nanomaterials, are inevitably released into aquatic environments, posing potential threats to aquatic organisms. Mytilus galloprovincialis is a bivalve species sensitive to changes in marine ecological environments, but there has been limited research on its toxicity response to ZnO NPs. Therefore, we selected M. galloprovincialis as the research subject and exposed them to 50 µg/L ZnO NPs for 96 h and 30 days to determine the dissolution of ZnO NPs in seawater and their distribution in M. galloprovincialis. The toxicity of ZnO NPs in M. galloprovincialis was then evaluated through gene expression, tissue pathology, and cellular immune response. The results showed that ZnO NPs could enrich Zn in various tissues of the mussel, in the order of gills > hepatopancreas > adductor muscle > mantle. Seven immune-related genes including four heat shock protein genes (HSPA12A, sHSP24.1, sHSP22, TCTP) and three apoptotic genes (Ras, p63 and Bcl-2) were altered to varying degrees. There was a downward trend in lysosomal membrane stability of M. galloprovincialis after exposure to ZnO NPs for 96 h and 30 days, while ROS and apoptosis rates increased significantly. Furthermore, the seven genes, apoptosis, LMS, and ROS were dependent on exposure time, treatment, and their interaction. Histopathological damage included disorganisation of hepatopancreas epithelial cells, gill filament swelling, and contraction of blood sinuses. These results indicated that ZnO NPs exerted toxicity in M. galloprovincialis, affecting the immune system, resulting in changes in the expression of immune-related genes and ultimately leading to histopathological changes. Our research findings could contribute to systematically understand the impact of ZnO NPs on bivalves in aquatic environments and provide a theoretical basis for marine pollution assessment.

Effect of propolis applied to goat kids at weaning period on heat shock protein genes

Weaning is a stressful period that affects the welfare of goat kids. During this period, goat kids lose weight due to reduced food intake, and they become susceptible to viral and bacterial infections. In recent years, many studies have been carried out on more natural and organic additives to prevent the loss of offspring in goat breeding and to promote their growth and development by limiting the use of antibiotics. Propolis has potent immunomodulatory, antioxidant, and anti-inflammatory properties. We aimed to investigate the effects of propolis on weaning stress in this study. The expression levels of the molecular chaperones which modulate the cellular stress response HSP27, HSP60, and HSP70 were examined. The Saanen kids were divided into propolis-treated (n=10) and control (non-propolis treated; n=10) groups. The propolis-treated group was given 0.4 cc of propolis once daily for 2 weeks. Expression levels were calculated by 2 -ΔΔCt using the Pfaffl method in the sample taken on the day after weaning and at the end of the 2-week propolis application. During weaning stress in kids, the expression levels of HSP27 and HSP60 were increased by 2-fold or more while HSP70 was upregulated by 3.57 fold. When 0.4 cc of propolis was administered to kids under weaning stress, there was a statistically significant down-regulation of 0.99-fold in the HSP27, 1.53-fold in the HSP60 level and 2.61-fold in HSP70 level. Our study showed that propolis application reduced stress protein levels during weaning stress.

Genetics and ontogeny are key factors influencing thermal resilience in a culturally and economically important bivalve

Increasing seawater temperatures coupled with more intense and frequent heatwaves pose an increasing threat to marine species. In this study, the New Zealand green-lipped mussel, Perna canaliculus, was used to investigate the effect of genetics and ontogeny on thermal resilience. The culturally and economically significant mussel P. canaliculus (Gmelin, 1971) has been selectively-bred in New Zealand for two decades, making it a unique biological resource to investigate genetic interactions in a temperate bivalve species. Six selectively-bred full sibling families and four different ages, from early juveniles (6, 8, 10 weeks post-fertilisation) to sub-adults (52 weeks post-fertilisation), were used for experimentation. At each age, each family was exposed to a three-hour heat challenge, followed by recovery, and survival assessments. The shell lengths of live and dead juvenile mussels were also measured. Gill tissue samples from sub-adults were collected after the thermal challenge to quantify the 70 kDa heat shock protein gene (hsp70). Results showed that genetics, ontogeny and size influence thermal resilience in P. canaliculus, with LT50 values ranging between 31.3 and 34.4 °C for all studied families and ages. Juveniles showed greater thermotolerance compared to sub-adults, while the largest individuals within each family/age class tended to be more heat sensitive than their siblings. Sub-adults differentially upregulated hsp70 in a pattern that correlated with net family survival following heat challenge, reinforcing the perceived role of inducible HSP70 protein in molluscs. This study provides insights into the complex interactions of age and genotype in determining heat tolerance of a key mussel species. As marine temperatures increase, equally complex selection pressure responses may therefore occur. Future research should focus on transcriptomic and genomic approaches for key species such as P. canaliculus to further understand and predict the effect of genetic variation and ontogeny on their survival in the context of climate change.

Genome-wide identification of the HSP70 genes in Pacific oyster Magallana gigas and their response to heat stress.

Heat shock protein 70 (HSP70), the most prominent and well-characterized stress protein in animals, plays an important role in assisting animals in responding to various adverse conditions. In the present study, a total of 113 HSP70 gene family members were identified in the updated genome of Magallana gigas (designated MgHSP70) (previously known as Crassostrea gigas). There were 75, 12, 11, and 8 HSP70s located in the cytoplasm, nucleus, mitochondria, and endoplasmic reticulum, respectively, and 7 HSP70s were located in both the nucleus and cytoplasm. Among 113 MgHSP70 genes, 107 were unevenly distributed in 8chromosomes of M. gigas with the greatest number in chromosome 07 (61 genes, 57.01%). The MgHSP70 gene family members were mainly assigned intofive clusters, among which the HSPa12 subfamily underwent lineage-specific expansion, consisting of 89 members. A total of 68 MgHSP70 genes (60.18%) were tandemly duplicated and formed 30 gene pairs, among which 14 gene pairs were under strong positive selection. In general, the expression of MgHSP70s was tissue-specific, with the highest expression in labial palp and gilland the lowest expression in adductor muscle and hemocytes. There were 35, 31, and 47 significantly upregulated genes at 6, 12, and 24h after heat shock treatment (28°C), respectively. The expression patterns of different tandemly duplicated genes exhibited distinct characteristics aftershock treatment, indicating that these genes may have different functions. Nevertheless, genes within the same tandemly duplicated group exhibit similar expression patterns. Most of the tandemly duplicated HSP70 gene pairs showed the highest expression levels at 24h. This study provides a comprehensive description of the MgHSP70 gene family in M. gigas and offers valuable insights into the functions of HSP70 in the mollusc adaptation of oysters to environmental stress.

Function of unconventional T cells in oral lichen planus revealed by single-cell RNA sequencing.

We intended to map the single-cell profile of OLP, explore the molecular characteristics of unconventional T cells in OLP tissues. Buccal mucosa samples from OLP patients and healthy individuals were used to prepare single-cell suspension. Single-cell RNA sequencing was used to analyze the proportion of all the cells, and the molecular characteristics of unconventional T cells. Immunohistochemical staining was used to detect the expression of unconventional T cells marker genes. The cell clusters from buccal mucosa were categorized into immune cells, fibroblasts, endothelial cells, and epithelial cells. Unconventional T cells with phenotype of CD247+TRDC+NCAM1+ were identified. Immunohistochemical staining revealed higher expression of unconventional T cell marker genes in OLP tissue, predominantly in the lamina propria. In OLP, unconventional T cells are in a unique stress response state, exhibited enhanced NF-κB signaling and apoptosis inhibition, enhanced heat shock protein genes expression, weakened cytotoxic function. A large number of ligand-receptor pairs were found between unconventional T cells and other cells, particularly with fibroblasts and endothelial cells. This study mapped the single-cell profile of OLP, delineated the molecular characteristics of unconventional T cells in OLP, and uncovered that these unconventional T cells are in a stress response state.

Comparative transcriptome analysis provides comprehensive insight into the molecular mechanisms of heat adaption in Plutella xylostella

Plutella xylostella is one of the most destructive pests for cruciferous vegetables, and is adaptability to different environmental stressors. However, we still know little about the molecular mechanisms of how P. xylostella adapt to thermal stress. Here, the comparative transcriptome analysis was conducted from the samples of control (27 °C, CK) and heat treatment (40 °C, 40 T) P. xylostella. The results showed 1253 genes were differentially expressed, with 624 and 629 genes up- and down-regulated respectively. The annotation analysis demonstrated that “Energy production and conversion”, “Protein processing in endoplasmic reticulum”, “Peroxisome” and “Tyrosine metabolism” pathways were significantly enriched. Additionally, we found the expression levels of heat shock protein genes (Hsps), cuticle related genes and mitochondrial genes were significantly up-regulated in 40 T insects, suggesting their vital roles in improving adaption to heat stress. Importantly, the SOD activity and MDA content of P. xylostella were both identified to be increased under high temperature stress, indicating the elevated antioxidant reactions might be involved in response to heat stress. In conclusion, the present study offered us an overview of gene expression changes after 40 °C treatments, and found some critical pathways and genes of P. xylostella might play the critical roles in resisting heat stress.

Genome-wide identification of gene families related to miRNA biogenesis in Mangifera indica L. and their possible role during heat stress.

Mango is a popular tropical fruit that requires quarantine hot water treatment (QHWT) for postharvest sanitation, which can cause abiotic stress. Plants have various defense mechanisms to cope with stress; miRNAs mainly regulate the expression of these defense responses. Proteins involved in the biogenesis of miRNAs include DICER-like (DCL), ARGONAUTE (AGO), HYPONASTIC LEAVES 1 (HYL1), SERRATE (SE), HUA ENHANCER1 (HEN1), HASTY (HST), and HEAT-SHOCK PROTEIN 90 (HSP90), among others. According to our analysis, the mango genome contains five DCL, thirteen AGO, six HYL, two SE, one HEN1, one HST, and five putative HSP90 genes. Gene structure prediction and domain identification indicate that sequences contain key domains for their respective gene families, including the RNase III domain in DCL and PAZ and PIWI domains for AGOs. In addition, phylogenetic analysis indicates the formation of clades that include the mango sequences and their respective orthologs in other flowering plant species, supporting the idea these are functional orthologs. The analysis of cis-regulatory elements of these genes allowed the identification of MYB, ABRE, GARE, MYC, and MeJA-responsive elements involved in stress responses. Gene expression analysis showed that most genes are induced between 3 to 6 h after QHWT, supporting the early role of miRNAs in stress response. Interestingly, our results suggest that mango rapidly induces the production of miRNAs after heat stress. This research will enable us to investigate further the regulation of gene expression and its effects on commercially cultivated fruits, such as mango, while maintaining sanitary standards.

Genome-Wide Identification and Characterization of RdHSP Genes Related to High Temperature in Rhododendron delavayi.

Heat shock proteins (HSPs) are molecular chaperones that play essential roles in plant development and in response to various environmental stresses. Understanding R. delavayi HSP genes is of great importance since R. delavayi is severely affected by heat stress. In the present study, a total of 76 RdHSP genes were identified in the R. delavayi genome, which were divided into five subfamilies based on molecular weight and domain composition. Analyses of the chromosome distribution, gene structure, and conserved motif of the RdHSP family genes were conducted using bioinformatics analysis methods. Gene duplication analysis showed that 15 and 8 RdHSP genes were obtained and retained from the WGD/segmental duplication and tandem duplication, respectively. Cis-element analysis revealed the importance of RdHSP genes in plant adaptations to the environment. Moreover, the expression patterns of RdHSP family genes were investigated in R. delavayi treated with high temperature based on our RNA-seq data, which were further verified by qRT-PCR. Further analysis revealed that nine candidate genes, including six RdHSP20 subfamily genes (RdHSP20.4, RdHSP20.8, RdHSP20.6, RdHSP20.3, RdHSP20.10, and RdHSP20.15) and three RdHSP70 subfamily genes (RdHSP70.15, RdHSP70.21, and RdHSP70.16), might be involved in enhancing the heat stress tolerance. The subcellular localization of two candidate RdHSP genes (RdHSP20.8 and RdHSP20.6) showed that two candidate RdHSPs were expressed and function in the chloroplast and nucleus, respectively. These results provide a basis for the functional characterization of HSP genes and investigations on the molecular mechanisms of heat stress response in R. delavayi.

Effects of heat shocks, heat waves, and sustained warming on solitary bees

Along with higher average temperatures, global climate change is expected to lead to more frequent and intense extreme heat events, and these different types of warming are likely to differ in their effects on bees. Although solitary bees comprise &amp;gt;75% of bee species, and despite their ecological and economic value as pollinators, a literature search revealed that only 8% of studies on bee responses to warming involve solitary bees. Here we review studies that have addressed how solitary bees are affected by three main types of warming that vary in magnitude and duration: heat shocks, heat waves, and sustained warming. We focus on direct physiological and behavioral effects of warming on solitary bees, rather than the underlying mechanisms. We find that heat shocks have received little attention in solitary bees both in terms of number of studies and relative to social bees, and all of those studies examine the effects of heat shocks on a single genus, Megachile. This work has shown that heat-shocked eggs, larvae, and pupae tend to upregulate heat shock protein genes, while heat shock at the adult stage can increase mortality in male bees, potentially altering population sex ratios. We find that solitary bee responses to heat waves have received even less study, but the few studies suggest that these events can increase larval mortality and slow development time, and that bees may not be able to physiologically acclimate to heat wave conditions by increasing their critical thermal maxima. Finally, sustained warming, which has been relatively well-studied in solitary bees, can speed development rate, reduce body mass, increase mortality, and alter foraging behavior. Our review reveals knowledge gaps in the effects of heat shocks and heat waves on solitary bees and, more broadly, in the responses of unmanaged solitary bees to warming. To improve our ability to anticipate the consequences of climate change for these critical pollinators, we encourage research on solitary bee thermal responses that examines short-term, extreme warming and incorporates greater ecological realism and complexity.

Genome-wide identification and expression analysis of the heat shock protein gene superfamily in Hylurgus ligniperda

Hylurgus ligniperda belongs to Hylurgus Latreille, Curculionidae, Coleoptera. It primarily damages the base and roots of the trunk of pine plants. Short-term treatment at 42 °C can damage Hylurgus ligniperda; therefore, temperature is a vital factor limiting its spread. Heat shock proteins (HSPs) can protect, remove, and repair proteins to help H. ligniperda withstand high temperatures. However, information on HSP genes in H. ligniperda remains limited. In the study, we considered H. ligniperda as the focus of research and identified 56 HligHSP genes at the genome-wide level. These genes were mapped to the cytoplasm or nucleus. An identical subfamily exhibited a closely similar distribution of conserved domains. Combined with the transcriptome data collected in previous studies, we screened six candidate genes, namely HligsHSP-3, HligsHSP-4, HligHSP60-16, HligHSP70-3, HligHSP70-4, and HligHSP90-1, which are specifically expressed during different high-temperature treatments. A quantitative polymerase chain reaction was performed to measure the expression of these six HligHSPs in seven temperature treatment conditions. These genes may be involved in the heat resistance mechanism in adults. Our findings provided a foundation for further studying the heat resistance mechanism in H. ligniperda.

JRK binds satellite III DNA and is necessary for the heat shock response.

JRK is a DNA-binding protein of the pogo superfamily of transposons, which includes the well-known centromere binding protein B (CENP-B). Jrk null mice exhibit epilepsy, and growth and reproductive disorders, consistent with its relatively high expression in the brain and reproductive tissues. Human JRK DNA variants and gene expression levels are implicated in cancers and neuropsychiatric disorders. JRK protein modulates β-catenin-TCF activity but little is known of its cellular functions. Based on its homology to CENP-B, we determined whether JRK binds centromeric or other satellite DNAs. We show that human JRK binds satellite III DNA, which is abundant at the chromosome 9q12 juxtacentromeric region and on Yq12, both sites of nuclear stress body assembly. Human JRK-GFP overexpressed in HeLa cells strongly localises to 9q12. Using an anti-JRK antiserum we show that endogenous JRK co-localises with a subset of centromeres in non-stressed cells, and with heat shock factor 1 following heat shock. Knockdown of JRK in HeLa cells proportionately reduces heat shock protein gene expression in heat-shocked cells. A role for JRK in regulating the heat shock response is consistent with the mouse Jrk null phenotype and suggests that human JRK may act as a modifier of diseases with a cellular stress component.

Effect of seven days heat stress on feed and water intake, milk characteristics, blood parameters, physiological indicators, and gene expression in Holstein dairy cows

This study examined the effects of 7 days of heat stress on eight early lactating Holstein cows in climate-controlled chambers. The early lactating Holstein cows (42 ± 2 days in milk, 29.27 ± 0.38 kg/day milk yield, 1.21 ± 0.05 parity) were subjected to two 14-day periods, each consisting of 7 days of adaptation and 7 days of heat stress. Conditions were set to 22 °C and 50% humidity during adaptation, followed by heat stress periods with low-temperature, low-humidity (LTLH, 71 THI) and high-temperature, high-humidity (HTHH, 86 THI) treatments. Data from the last 7 days were analyzed using a mixed procedure in SAS. In the study, the HTHH group displayed marked physiological and biochemical changes on 14 days of heat stress exposure compared to the LTLH group. Firstly, the HTHH group's dry matter intake decreased by approximately 12% while their water intake increased by about 23%. Secondly, both milk yield and milk protein production in the HTHH group decreased by 10% and 20%, respectively. Thirdly, there was a reduction in white blood cells, hemoglobin, mean corpuscular hemoglobin, and platelets in the HTHH group, with concurrent increases in glucose, non-esterified fatty acids, and albumin concentrations. Additionally, the HTHH group exhibited elevated plasma concentrations of cortisol and haptoglobin. Moreover, the gene expression of heat shock protein 70 and heat shock protein 90 was significantly upregulated in the HTHH group's peripheral blood mononuclear cells. Lastly, key physiological indicators such as rectal temperature, heart rate, and skin temperature showed substantial elevations in the HTHH group. Considering the enormous negative effects observed in the analyzed blood metabolites, milk yield and compositions, and heat shock protein gene expression, early lactating Holstein cows were found to be more vulnerable to HTHH than LTLH over a 7 days exposure to heat stress.

PeHSFA3 is essential for the heat-stress response of Populus × euramericana

Heat shock transcription factors (HSFs) are crucial regulators in the responses of plants to heat stress, but their mechanisms of action in trees are unclear. We identified an HSFA gene, PeHSFA3, in Populus × euramericana (Populus deltoides × Populus nigra), which was highly responsive to heat. Its protein product was in both the cytoplasm and the nucleus. Stably overexpressing PeHSFA3 in poplar enhanced heat tolerance, improved photosynthetic capacity, increased antioxidant enzyme activities, reduced reactive oxygen species (ROS) levels, and upregulated antioxidant-related genes such as SODF2, SODCP, PER46–1, PER46–2, and PER31 under heat stress. Conversely, suppressing the PeHSFA3 gene by Virus-induced gene silencing (VIGS) in Populus × euramericana decreased its heat tolerance. qRT-PCR analysis demonstrated that PeHSFA3 enhances the expression of several heat-protective genes, including small heat-shock protein genes (HSP22C, HSP21–5, HSP18.1, HSP16A, HSP16B, HSP26.5–1, and HSP26.5–2) and ATG8. Transcriptomic analysis suggested that the activation of glutathione metabolic pathways contributes to PeHSFA3-mediated heat tolerance. Our findings reveal that PeHSFA3 mediates resistance to heat stress in poplar and could be used to enhance the thermotolerance of poplar.

Early Socialization Triggered ROS-Mediated Activation of Canonical NF-κB Pathway Leading to Inflammation of Spleen in Suckling Piglets

Early socialization during lactation is advocated as a feeding strategy to reduce the weaning stress of piglets. However, early socialization has often been accompanied by more frequent aggression between individuals, and its effect on the immune system of piglets has yet to be evaluated. In this study, 89 piglets were raised separately under conventional feeding and early socialization environments. Based on differences in the aggressive behavior of the piglets in different environments during lactation, we further investigated the effects of early socialization on oxidative stress in the spleen of the piglets and the inflammatory responses involved in the canonical nuclear factor kappa-B (NF-κB) signaling pathway. The results revealed that early socialization led to a higher aggression level between individuals (p &lt; 0.01), increased malondialdehyde (MDA) and H2O2 levels and inducible nitric oxide synthase (iNOS) activity, and inhibited glutathione (GSH) levels and the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX) in the piglet spleens (p &lt; 0.05). The mRNA expression levels of the protein kinase A (PKA), inhibitor of kappa B kinase-α (IKK-α), inhibitor of kappa B kinase-β (IKK-β), inhibitor of NF-κB-α (IκB-α), NF-κB(p65), interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX2), iNOS, and heat shock protein (HSP) genes were significantly up-regulated, as well as the protein levels of P-p65, IKK-β, P-IkB-α, pro-IL-1β, and TNF-α. In summary, early socialization caused oxidative stress and inflammatory responses in the spleen of the piglets by inducing ROS production and the activation of the canonical NF-κB pathway. Our study revealed that early socialization significantly increased the ROS level in the piglet spleens and activated the canonical NF-κB signaling pathway, which induced a high expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and COX2) and HSP genes regulated by NF-κB signaling, leading to oxidative stress and the inflammatory response.

Analysis of heat-shock protein genes and their expression in Lactiplantibacillus plantarum SU-KC1a

Abstract. Julyanto CMP, Dosan R, Victor H, Sugata M, Tan TJ. 2024. Analysis of heat?shock protein genes and their expression in Lactiplantibacillus plantarum SU-KC1a. Biodiversitas 25: 2494-2499. Lactiplantibacillus plantarum is lactic acid bacteria commonly used as probiotics. Throughout the manufacturing and processing stages for incorporation into functional food products, probiotics encounter different stress conditions, including heat stress, which can affect their viability. In this study, the ability of L. plantarum SU-KC1a to withstand heat stress conditions was assessed by measuring cell viability after incubation at 37, 42, 47, and 52? for one hour. Additionally, the growth of SU-KC1a in liquid cultures under various high temperatures was monitored hourly for three hours. Total RNA was extracted from each culture, converted into cDNA, and amplified using hsp3 gene-specific primers. The results indicated a slower growth rate of SU-KC1a at higher temperatures, with 42? as the maximum temperature supporting growth. Visualization of hsp3 amplicon showed increased intensity following incubation at higher temperatures, indicating upregulation of hsp3 gene expression. To identify the presence of heat stress-related genes, whole genome sequence annotation data of SU-KC1a was compared to that of L. plantarum SK151, F75, and WCFS1. Lactiplantibacillus plantarum was found to possess two systems involved in protein repair in response to heat stress damage: chaperone and protease. In summary, L. plantarum SU-KC1a harbors heat shock protein genes, and their expression is upregulated following exposure to high temperatures.

Molecular characterization and expression analysis of six small heat shock protein genes in Trogoderma granarium during cold and starvation-induced larval diapause

The khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae), is a highly invasive pest infesting stored grains, cereal products, and is recognised as a quarantine pest in many countries. Various stressors can instigate prolonged diapause among insect larvae, increasing the challenge of managing infestation. When the environmental conditions become conducive to development, diapause ceases, and the larva progresses on its developmental course. Members of the Heat Shock Protein (HSP) family play a significant role in protecting insects from the impacts of environmental stressors. In this study, six small heat shock proteins (TsHSP19.8, TsHSP20.5, TsHSP20.6, TsHSP20.8, TsHSP21.2, and TsHSP21.3) were identified and characterized in T. granarium. Transcript abundances of TsHSPs were analysed across distinct phases of larval diapause induced by both cold exposure and starvation stress treatments, alongside various developmental stages. The gene expression patterns of TsHSP20.5 and TsHSP21.2 exhibited peak mRNA levels during pre-diapause following the cold treatment, while the greatest abundances of TsHSP19.8, TsHSP20.6, TsHSP20.8, and TsHSP21.3 were detected in the diapause phases. The highest mRNA abundances of TsHSP20.5, TsHSP20.8, TsHSP21.2, and TsHSP21.3 were observed during the diapause phase following the starvation treatment, while it was noted on the first day of the post-diapause phase for TsHSP19.8 and TsHSP20.6. The remarkable levels of TsHSPs during diapause might imply their putative role in cold and starvation survival. Significant expression of TsHSP19.8, TsHSP20.6, TsHSP20.8, and TsHSP21.3 transcripts was observed in adults, suggesting their potential involvement in developmental processes. Understanding the genetic mechanisms involved in the diapause physiology of T. granarium could offer additional insights for the integrated management of this pest.

MaHsf24, a novel negative modulator, regulates cold tolerance in banana fruits by repressing the expression of HSPs and antioxidant enzyme genes.

Transcriptional regulation mechanisms underlying chilling injury (CI) development have been widely investigated in model plants and cold-sensitive fruits, such as banana (Musa acuminata). However, unlike the well-known NAC and WRKY transcription factors (TFs), the function and deciphering mechanism of heat shock factors (HSFs) involving in cold response are still fragmented. Here, we showed that hot water treatment (HWT) alleviated CI in harvested banana fruits accomplishing with reduced reactive oxygen species (ROS) accumulation and increased antioxidant enzyme activities. A cold-inducible but HWT-inhibited HSF, MaHsf24, was identified. Using DNA affinity purification sequencing (DAP-seq) combined with RNA-seq analyses, we found three heat shock protein (HSP) genes (MaHSP23.6, MaHSP70-1.1 and MaHSP70-1.2) and three antioxidant enzyme genes (MaAPX1, MaMDAR4 and MaGSTZ1) were the potential targets of MaHsf24. Subsequent electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) and dual-luciferase reporter (DLR) analyses demonstrated that MaHsf24 repressed the transcription of these six targets via directly binding to their promoters. Moreover, stably overexpressing MaHsf24 in tomatoes increased cold sensitivity by suppressing the expressions of HSPs and antioxidant enzyme genes, while HWT could recover cold tolerance, maintaining higher levels of HSPs and antioxidant enzyme genes, and activities of antioxidant enzymes. In contrast, transiently silencing MaHsf24 by virus-induced gene silencing (VIGS) in banana peels conferred cold resistance with the upregulation of MaHSPs and antioxidant enzyme genes. Collectively, our findings support the negative role of MaHsf24 in cold tolerance, and unravel a novel regulatory network controlling bananas CI occurrence, concerning MaHsf24-exerted inhibition of MaHSPs and antioxidant enzyme genes.

Climate change induces shifts in coastal Baltic Sea surface water microorganism stress and photosynthesis gene expression.

The world's oceans are challenged by climate change linked warming with typically highly populated coastal areas being particularly susceptible to these effects. Many studies of climate change on the marine environment use large, short-term temperature manipulations that neglect factors such as long-term adaptation and seasonal cycles. In this study, a Baltic Sea 'heated' bay influenced by thermal discharge since the 1970s from a nuclear reactor (in relation to an unaffected nearby 'control' bay) was used to investigate how elevated temperature impacts surface water microbial communities and activities. 16S rRNA gene amplicon based microbial diversity and population structure showed no difference in alpha diversity in surface water microbial communities, while the beta diversity showed a dissimilarity between the bays. Amplicon sequencing variant relative abundances between the bays showed statistically higher values for, e.g., Ilumatobacteraceae and Burkholderiaceae in the heated and control bays, respectively. RNA transcript-derived activities followed a similar pattern in alpha and beta diversity with no effect on Shannon's H diversity but a significant difference in the beta diversity between the bays. The RNA data further showed more elevated transcript counts assigned to stress related genes in the heated bay that included heat shock protein genes dnaKJ, the co-chaperonin groS, and the nucleotide exchange factor heat shock protein grpE. The RNA data also showed elevated oxidative phosphorylation transcripts in the heated (e.g., atpHG) compared to control (e.g., atpAEFB) bay. Furthermore, genes related to photosynthesis had generally higher transcript numbers in the control bay, such as photosystem I (psaAC) and II genes (psbABCEH). These increased stress gene responses in the heated bay will likely have additional cascading effects on marine carbon cycling and ecosystem services.

The class B heat shock factor RcHsf17 from Rosa chinensis enhances basal thermotolerance in Rosa rugosa

Rosa rugosa, a species of notable economic importance, faces cultivation challenges attributed to its vulnerability to thermal stress. In this study, we report the molecular cloning and functional characterization of RcHsf17, a heat shock transcription factor (Hsf) from group B2, derived from the stress-resilient cultivar of Chinese rose, R. chinensis ‘Slater's Crimson China’. RcHsf17, which localizes to the nucleus, is responsive to heat stress and various stress-related stimuli, such as abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), and hydrogen peroxide (H2O2). The overexpression of RcHsf17 in transgenic Arabidopsis thaliana and R. rugosa significantly enhanced their thermotolerance. In R. rugosa, overexpression of RcHsf17 led to cell wall thickening, sustained photosynthetic efficiency, and decreased levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Transcriptome analysis revealed that RcHsf17 upregulated the expression of other Hsfs such as HsfA3 and HsfB1, small heat shock protein genes (sHSPs), and genes involved in cell wall biosynthesis under normal conditions. Under heat stress condition, there is a significant upregulation of HsfA6B, gene associated with photosynthesis, and genes responsible for encoding antioxidant enzymes. These findings collectively emphasize the pivotal role of RcHsf17 in conferring thermotolerance to R. rugosa, through mechanisms that include strengthening cell wall integrity, alleviating heat-induced photosynthetic decline, and enhancing the efficiency of ROS detoxification pathways.