Temperature Stress Conditions Research Articles

Impact of high temperature and drought stress on the microbial community in wolf spiders

High temperatures and drought present significant abiotic challenges that can limit the survival of many arthropods, including wolf spiders, which are ectothermic and play a crucial role in controlling pest populations. However, the impact of these stress factors on the microbiota of spiders remains poorly understood. In this study, we utilized 16 S rRNA gene sequencing to explore the diversity and composition of bacterial communities within Pardosa pseudoannulata under conditions of high temperature and drought stress. We found that Firmicutes, Bacteroidetes, and Proteobacteria were the predominant bacterial phyla present. Analyses of alpha diversity indicated an increase in bacterial diversity under combined stress conditions, as reflected by various diversity indices such as Ace, Chao1, Shannon, and Simpson. Furthermore, co-occurrence network analysis highlighted intricate interactions among the microbial taxa (e.g., Enterobacter, Chitinophaga, and Eubacterium), revealing the adaptive complexity of the spider's microbiome to environmental stress. Functional prediction analysis suggested that combined stress conditions might enhance key metabolic pathways, particularly those related to oxidative phosphorylation and amino acid metabolism. Using Random Forest analysis, we determined that changes in three heat shock proteins were largely attributed to variations in bacterial communities, with Firmicutes being notably influential. Collectively, this in-depth analysis offers novel insights into the responses of microbial communities within spider microbiomes to combined abiotic stresses, providing valuable information for understanding extreme climate impacts and informing ecological management strategies.

Foliar spraying with amino acids and their chitosan nanocomposites as promising way to alleviate abiotic stress in iceberg lettuce grown at different temperatures

We analyzed the effects of foliar spraying with amino acids, chitosan (CHS) and nanocomposites (NCs) of chitosan with the amino acids proline, l-cysteine and glycine betaine (CHS-Pro NCs; CHS-Cys NCs, CHS-GB NCs, respectively) on the changes in the physiological and biochemical parameters of iceberg lettuce grown at the control temperature (20 °C) and under chilling conditions (4 °C). The physicochemical parameters of the phospholipid monolayers (PLs) extracted from plants showed the effects of the treatments on the properties of the monolayers, namely, the packing density and flexibility. We observed increased accumulation of proline at 4 °C, and differences in the concentrations of sugars in most of the analyzed variants were a consequence of the lowered temperature and/or the use of organic compounds. A temperature of 4 °C caused a significant increase in the l-ascorbic acid level compared with that at 20 °C. Differences were also found in glutathione (GSH) content depending on the temperature and treatment with the tested organic compounds. CHS NCs loaded with Pro and GB were effective at increasing the amount of phenols under stress temperature conditions. We noted that a significant increase in the antioxidant activity of plants at 4 °C occurred after priming with Cys, CHS-Cys NCs, Pro and CHS-Pro NCs, and the CHS nanocomposites were more effective in this respect. Both low-temperature stress and foliar spraying of lettuce with various organic compounds caused changes in the activity of antioxidant enzymes. Two forms of dismutase (SOD), iron superoxide dismutase (FeSOD) and copper/zinc superoxide dismutase (Cu/ZnSOD), were identified in extracts from the leaves of iceberg lettuce seedlings. The application of the tested organic compounds, alone or in combination with CHS, increased the amount of malondialdehyde (MDA) in plants grown under controlled temperature conditions. Chilling caused an increase in the content of MDA, but some organic compounds mitigated the impact of low temperature. Compared with that of plants subjected to 20 °C, the fresh weight of plants exposed to chilling decreased. However, the tested compounds caused a decrease in fresh weight at 4 °C compared with the corresponding control samples. An interesting exception was the use of Cys, for which the difference in the fresh weight of plants grown at 20 °C and 4 °C was not statistically significant. After Cys application, the dry weight of the chilled plants was greater than that of the chilled control plants but was also greater than that of the other treated plants in this group. To our knowledge, this is the first report demonstrating that engineered chitosan–amino acid nanocomposites could be applied as innovative protective agents to mitigate the effects of chilling stress in crop plants.

Exploring the role of 28-homobrassinolide in regulation of temperature induced clastogenic aberrations and sugar metabolism of Brassica juncea L.

The present research primarily focuses on Brassica juncea's physiological and cytological responses to low and high temperature stress at 4 °C and 44 °C respectively, along with elucidating the protective role of 28-Homobrassinolide (28-homoBL). Cytological investigations performed in floral buds of Brassica juncea L. under temperature (24, 4, 44 °C) stress conditions depict the presence of some abnormalities associated with cytomixis such as chromosome stickiness or agglutination, pycnotic nature of chromatin, irregularities in spindle formation, disoriented chromatins, and non-synchronous chromatin material condensation in Brassicaceae family that subsisted at diploid level (2n = 36). Spindle abnormalities produce various size pollen grains such as sporads micronuclei at some stages of microsporogenesis, polyads, triads, dyads that irrupted the productiveness of pollen grains. Furthermore, sugars play an imperative role in protecting plants under stress besides being energy sources. Therefore, the present study revealed accumulation of total soluble sugars (TSS), with 28-homoBL treatment which pinpoints protective role of 28-homoBL under temperature stress. Sugar profiling was done by using high-performance liquid chromatography (HPLC) which helped in analyzing different sugars both quantitatively and qualitatively under 28-homoBL and temperature stress conditions. The results indicate that the 28-homoBL treatment substantially enhances plant tolerance to heat stress, as evident by higher mitotic indices, fewer chromosomal abnormalities, and significantly more sugar accumulation. The findings of the study acknowledge the potential of 28-homoBL in inducing temperature stress tolerance in B. juncea along with improving the metabolic stability thereby implying application of 28-homoBL in crop strengthening under variable temperature conditions.

The stability of phycocyanin extracted from Arthrospira platensis against osmotic, acid, and temperature stress conditions

The main problem of using natural blue pigment of phycocyanin produced by Arthrospira platensis (spirulina) is its instability in the food matrix because of environmental stress. This study aimed to investigate the stability of phycocyanin under simulated conditions in food formulations against osmotic, acid, and temperature stress conditions. Thermal degradation constant (Dc) and half-life (t½) of phycocyanin extracted from Arthrospira platensis were analyzed using response surface methodology followed by a first-order kinetic reaction. The stability of phycocyanin was assessed under various temperature (50–98ºC), NaCl (0–2% w/w), and pH (4–7) values. Results showed that the stability of phycocyanin extracted from Arthrospira platensis is high at neutral pH and concentration of 1% (w/w) NaCl. The stability decreased with increase in temperature at ≥75°C. The highest stability of phycocyanin (the lowest Dc and the maximum t½ were 0.011 min-1 and 54.03 min, respectively) was achieved at 66.89°C, pH = 6.6, and NaCl of 0.40% w/w. According to processing conditions, content of phycocyanin required for a food matrix is successfully calculated by the response surface method. This research showed that phycocyanin is stable at thermal shock in a neutral pH medium and low content of NaCl (0.40% w/w).

Development and validation of stability-indicating method of etrasimod by HPLC/DAD/MS/MS technique with greenness profiling

Etrasimod, a novel selective sphingosine-1-phosphate receptor modulator, was recently approved by the U.S. Food and Drug Administration and the European Medicinal Agency for the treatment of moderately to severely active ulcerative colitis in adults. In this research, the forced degradation study as an integral part of drug product and packaging development, which generates data on degradation mechanisms, is published. The development and validation of the stability-indicating method using a superior high-performance liquid chromatography technique coupled with a diode array detector and tandem mass spectrometer was performed to support the forced degradation study and monitor the formation of degradation products. Etrasimod demonstrated good stability under elevated temperature and basic stress conditions, while acidic hydrolysis, oxidative, and photolytic degradation produced eight degradation products. The knowledge of degradation products will be useful in the long-term stability study for establishing the acceptance criteria of etrasimod as a drug substance and dosage form during quality control and stability assessment. The eco-friendliness of the developed forced degradation procedure was evaluated using various greenness appraisal tools. The green metric tools showed that the forced degradation procedure obeys eco-friendly conditions.

Intraspecific variation in responses to extreme and moderate temperature stress in the wild species, Solanum carolinense (Solanaceae).

Adaptation or acclimation to local temperature regimes has often been used as a proxy for predicting how plant populations will respond to impending novel conditions driven by human-caused climate change. To understand how plants may successfully respond to increasing air temperatures (extreme and moderate) in the future, we explored how temperature tolerance traits differ in populations of Solanum carolinense from northern (MN) and southern (TX) regions of the continental USA in a two-experiment study. In the first experiment, we compared the heat and cold tolerance in vegetative (sporophyte) and reproductive (male gametophyte) traits. In the second experiment, we studied if long-term heat influences plant development by examining how development in moderate heat affected reproductive structures and reproductive success. We found that temperature sensitivity differed between southern populations, which regularly experience extreme heat, and northern populations which do not. In contrast to our expectations, northern populations appeared more heat-tolerant than southern populations for vegetative traits such as chlorophyll stability and reproductive traits such as pollen germination. Our results are consistent with a heat-avoidance, rather than tolerance mechanism to mitigate extreme heat during pollen germination. In the second experiment, plants developing under the moderate heat treatment had significantly smaller reproductive structures and reduced seed production (27% fewer seeds on average than in the control treatment). Reproductive structures that developed in moderate heat were also reduced in size, particularly in the northern populations relative to populations from the south. We conclude that rising temperatures have the potential to incur substantial negative consequences for the reproductive success of individuals in this species and that some populations already mitigate stressful temperature conditions through phenotypic plasticity.

Design Implementation of a Logger-Based Digital Thermometer

LM35 temperature sensor is a widely used 3-terminal sensor known for its precise temperature measurement capabilities without the need for external calibration circuitry. With a sensitivity of 10mV/°C, it operates within a temperature range of -55°C to 150°C, with an error margin of 0.5°C. This paper presents the development and testing of a logger-based digital thermometer using LM35 sensor, integrated with a dsPIC30F4013 digital signal processing controller, an LCD, a real-time clock (DS1307), and other essential components. The methodology involves the analysis of the sensor's sensitivity and its response under both stress and non-stress temperature conditions. The findings indicate that the sensor's output closely follows the theoretical sensitivity, with a negligible offset, ensuring accurate temperature measurements. Comparative analysis with a standard laboratory thermometer suggests that the digital sensor provides superior precision, with an error margin of ±0.01°C. The results confirm LM35's suitability for accurate ambient temperature monitoring and validation of environmental temperature fluctuations.

Enhancement in electrical properties of dual-active-layer amorphous SiZnSnO/SiInZnO thin film transistors

Bi-layer thin film transistors (TFTs) have been fabricated with a channel structure comprising a dielectric layer, a semiconducting amorphous-Si-In-Zn-O (a-SIZO) layer, and a semiconducting amorphous-Si-Zn-Sn-O (a-SZTO) layer, aiming to improve field effect mobility and stability. These films were deposited using RF sputtering at room temperature. The TFTs with a bottom gate top contact, processed at 500 °C, exhibited high mobilities (>32 cm2 V−1 s−1) along with a current on/off ratio of approximately 108 and a subthreshold swing (SS) value below 0.5 V decade−1 primarily because of reduced trap density and presence of highly conducting ultrathin a-SIZO layer. Furthermore, the bi-layer TFTs demonstrated notable stability under negative and positive bias temperature stress conditions.

Association Analysis in Rice Germplasm Lines for High Temperature Tolerance

Background: Relative to other abiotic factors like salinity and drought, breeding rice cultivars that are resistant of high temperatures has received a little attention. The purpose of this study is to find parents and genotypes that are suitable for utilizing in high temperature stress breeding program. Methods: The genetic material for this study comprised of a set of germplasm collections (293 accessions) from different sources such as International Rice Research Institute germplasm. These populations were raised at the Department of Rice, Centre for Plant Breeding and Genetics, Tamil Nadu Agricultural University, Coimbatore. The Heat tolarance Indices (TOL) viz., Heat tolerance index (HTI) and Heat susceptibility index (HSI) and association analysis were estimated among germplasm lines for yield attributes under normal and high temperature stress condition. Result: Out of 293 genotypes, sixty nine genotypes significantly exceeded for both HTI and HSI. Association studies revealed that, single plant yield had highly significant and positive association with panicle exertion, number of total tillers per plant, number of productive tillers per plant, panicle length, number of filled grains per panicle and total dry matter production. Spikelet sterility and high temperature susceptibility index had highly significant and negative association with single plant yield. Number of productive tillers had significant and positive correlation with total dry matter production, panicle length, number of filled grains per panicle, days to maturity, hundred grain weight, grain breadth and high temperature susceptible index. Hence these traits may be considered as selection indices for yield and high temperature tolerance improvement programme. Hence it may be possible to combine single plant yield and heat susceptibility index by specific breeding programme for high temperature tolerance in rice.

Identification of CEP peptides encoded by the tobacco (Nicotiana tabacum) genome and characterization of their roles in osmotic and salt stress responses

Members of the CEP (C-terminally Encoded Peptide) gene family have been shown to be involved in various developmental processes and stress responses in plants. In order to understand the roles of CEP peptides in stress response, a comprehensive bioinformatics approach was employed to identify NtCEP genes in tobacco (Nicotiana tabacum L.) and to analyze their potential roles in stress responses. Totally 21 NtCEP proteins were identified and categorized into two subgroups based on their CEP domains. Expression changes of the NtCEP genes in response to various abiotic stresses were analyzed via qRT-PCR and the results showed that a number of NtCEPs were significant up-regulated under drought, salinity, or temperature stress conditions. Furthermore, application of synthesized peptides derived from NtCEP5, NtCEP13, NtCEP14, and NtCEP17 enhanced plant tolerance to different salt stress treatments. NtCEP5, NtCEP9 and NtCEP14, and NtCEP17 peptides were able to promote osmotic tolerance of tobacco plants. The results from this study suggest that NtCEP peptides may serve as important signaling molecules in tobacco's response to abiotic stresses.

Variation in Physiological Traits of Blackgram (Vigna mungo L.) Genotypes under High Temperature Stress

Background: Blackgram is one of the important short duration pulse crop which is sensitive to high temperatures. The rising global temperatures are threatening the yield of blackgram by altering the physiological processes at cellular level. Keeping this in view, the present investigation was carried out for better understanding of genotypic variability and the physiological mechanisms governing heat stress tolerance which can help in identifying heat tolerant blackgram genotypes that can yield better under climate change scenarios. Methods: Thirty blackgram genotypes selected from temperature induction response technique were evaluated for physiological efficiency under natural high temperature conditions during summer, 2022 and 2023. Field experiment was conducted at Agricultural College Farm, Acharya N.G Ranga Agricultural University, Agricultural College, Bapatla. Physiological and yield parameters were recorded at flowering and the data were analyzed statistically and pooled. Result: Significant genetic variability was observed among the blackgram genotypes with respect to all the physiological traits under high temperature stress conditions. Among the 30 genotypes tested for thermotolerance, the genotypes TBG-129, PU-1804, LBG-1015, PU-31 and TBG-104 were found to withstand high temperature stress at reproductive stage by maintaining higher total chlorophyll, carotenoids, chlorophyll stability index, total biomass production, cooler canopies resulting into higher canopy temperature depression, less membrane injury index values and higher yield. Correlation analysis revealed a strong positive correlation for all the physiological traits with seed yield except membrane injury index under heat stress conditions. The PCA results revealed considerable variability among the traits accounting for 82.65% of total variability. The genotypes TBG-129, PU-1804, LBG-1015 and TBG-104 can be potentially used as donors in the breeding programmes for the development of heat tolerant genotypes.

Transcription and splicing variations of SR genes accompany with genome-wide accumulation of long-introns in pine

Most of mRNAs in Eukaryote were matured after the removal of introns in their pre-mRNA transcripts. Serine/arginine-rich (SR) proteins are a group of splicing regulators regulating the splicing processes globally. Expressions of SR proteins themselves were extensively regulated, at both transcription and splicing levels, under different environmental conditions, specially heat stress conditions. The pine genome is characterized by super-long and easily methylated introns in a large number of genes that derived from the extensive accumulation of transposons (TEs). Here, we identified and analyzed the phylogenetic characteristics of 24 SR proteins and their encoding genes from the pine genome. Then we explored transcription and pre-mRNA splicing expression patterns of SR genes in P. massoniana seedlings under normal and heat stress temperature conditions. Our results showed that the transcription patterns of SR genes in pine exhibited significant changes compared to other plant species, and these changes were not strictly correlated with the intron length and DNA methylation intensity of the SR genes. Interestingly, none of the long introns of SR genes underwent alternative splicing (AS) in our experiment. Furthermore, the intensity of AS regulation may be related to the potential DNA methylation intensity of SR genes. Taken together, this study explores for the first time the characteristics of significant variations in the transcription and splicing patterns of SR proteins in a plant species with an over-accumulation of super-long introns.

Effect of cyclic overload on deformation, crack growth behavior and crack initiation/growth lives of a C(T) specimen for 12Cr steel under creep-fatigue condition

ABSTRACT In this study, using a C(T) specimen for 12Cr steel used for a steam turbine rotor, crack growth tests under creep-fatigue conditions with and without overloading were conducted and the fundamental behaviours of deformation and crack growth were investigated experimentally. As a result, load line displacement was found to be suppressed under overload condition regardless of temperature and stress holding time. Crack initiation life under high temperature and low stress condition with overloading was delayed as compared with no overload condition. However, under low temperature and high stress condition with short stress holding time, crack initiation was accelerated remarkably. Furthermore, although crack growth life under creep-fatigue conditions with overloading was found to be shortened as compared with no overloading, the effect of overloading on crack growth life was decreased with an increase in stress holding time.

The Roles of Circadian Clock Genes in Plant Temperature Stress Responses.

Plants monitor day length and memorize changes in temperature signals throughout the day, creating circadian rhythms that support the timely control of physiological and metabolic processes. The DEHYDRATION-RESPONSE ELEMENT-BINDING PROTEIN 1/C-REPEAT BINDING FACTOR (DREB1/CBF) transcription factors are known as master regulators for the acquisition of cold stress tolerance, whereas PHYTOCHROME INTERACTING FACTOR 4 (PIF4) is involved in plant adaptation to heat stress through thermomorphogenesis. Recent studies have shown that circadian clock genes control plant responses to temperature. Temperature-responsive transcriptomes show a diurnal cycle and peak expression levels at specific times of throughout the day. Circadian clock genes play essential roles in allowing plants to maintain homeostasis by accommodating temperature changes within the normal temperature range or by altering protein properties and morphogenesis at the cellular level for plant survival and growth under temperature stress conditions. Recent studies revealed that the central oscillator genes CIRCADIAN CLOCK ASSOCIATED 1/LATE ELONGATED HYPOCOTYL (CCA1/LHY) and PSEUDO-RESPONSE REGULATOR5/7/9 (PRR5/7/9), as well as the EVENING COMPLEX (EC) genes REVEILLE4/REVEILLE8 (REV4/REV8), were involved in the DREB1 pathway of the cold signaling transcription factor and regulated the thermomorphogenesis gene PIF4. Further studies showed that another central oscillator, TIMING OF CAB EXPRESSION 1 (TOC1), and the regulatory protein ZEITLUPE (ZTL) are also involved. These studies led to attempts to utilize circadian clock genes for the acquisition of temperature-stress resistance in crops. In this review, we highlight circadian rhythm regulation and the clock genes involved in plant responses to temperature changes, as well as strategies for plant survival in a rapidly changing global climate.

Hormonal panel of various breeds of cattle under the conditions of temperature stress

The article gives the data on the content of a number of steroid and thyroid hormones in the population of Tagil, Suksun and Holstein cattle under the conditions of increasing environmental temperature resulting in moderate stress reaction, as well as on the changes in endocrine system after temperature stress. Changes in function of vascular glands were found showing high resistance of Tagil cattle, moderate resistance of Suksun cattle and low resistance of Holstein cattle to temperature stress. The most significant changes were noted in thyroid panel of Tagil cattle that was shown by increasing synthesis of iodothyronine while environmental temperature changed from high to normal values (during post-stress period). The breeds with lower resistance had the signs of deterioration of adaption mechanism expressed in stronger unbalance reaction of hormonal axis.

An interpretation of thermo-mechanical behaviour of peat under 1-D compression

The one-dimensional thermo-mechanical behaviour of two peats was investigated through consolidation tests, including incremental loadings and constant-rate-of-strain loadings, under a variety of temperature-stress conditions. As an appropriate expression of highly compressible materials, the natural strain was employed instead of conventional engineering strain, which eliminates the apparent stress dependency of the compression index and the coefficient of secondary compression. Test results suggest the applicability of an isotach approach to the two peats used in this study. The temperature dependency of the coefficient of secondary compression defined with natural strain, λ*α, is very small. All the results summarized on the effective stress-strain plane show that there exists a unique Normal Compression Line (NCL) corresponding to each combination of temperature and strain rate. The NCLs are parallel to each other. Finally, in an attempt to construct a constitutive model, the temperature dependency of the NCL was quantified by introducing a constant, λ*T. By using the two parameters, λ*α and λ*T, which describe the strain rate- and temperature-dependent characteristics, respectively, a 1-D thermo-mechanical behaviour observed in this study will be modelled in a simple manner.

The Response of Different Bread Wheat Genotypes (Triticum aestivum L.) to Temperature Changes

Increased temperature is a major yield declining factor in wheat production. Current study was intended to evaluate the impact of increased temperature on the pre and post reproductive major yield attributing biometrical traits through the use of stress tolerance indices and trait reduction ratio (%) of all the traits studied. A set of 27 genotypes including 2 check entries viz., Shriram 303 and HD 2967 was evaluated in two separate trials, timely sown (non-stress) and late sown (heat stress) conditions in RBD design with three replications during rabi 2020-21 at the agriculture research farm of Rama university, Kanpur. Significant dropping in major yield attributing traits have Such as grain yield/plant, grain yield/spike and tillers/plant exhibited 54.41%, 39.31 % and 34.22% reduction, respectively thought the estimation of trait reduction ration (%) specially for the trials conducted under the late sown (heat stress) condition as compared to the normal sown trial. Five stress tolerance indices viz., susceptibility index (HSI), mean productivity (MP), tolerance (TOL), heat tolerance index (STI) and trait stability index (TSI) have been estimated to find out the heat stress tolerance and susceptible genotypes for grain yield under high temperature stress conditions.
 Correlation estimates of different heat stress tolerance indices with grain yield in non-stress condition (timely sown) exhibited significant positive association with MP (0.766**), HTI (0.622**), TSI (0.414*), and TOL (0.284). Though, under heat stress condition (late sown), grain yield displayed a significant positive correlation with HTI (0.713**), MP (0.707**), TOL (0.517**), TSI (0.656**) and negative correlation with HSI (-0.608**). Through the correlation analysis estimates, four heat stress indices viz. HSI (heat susceptibility index), MP (mean productivity), STI (stress tolerance index) and TSI (trait stability index) have been used in assorting the heat tolerant wheat genotypes. Current estimates directed that 14 genotypes (PBW 152, HUW-206, WL-2, WL-8, WL-13, DBW-71, DBW-39, WH-1105, HUW-318, PBW-154, ALWL-5, WL-14, PBW-502 and Shriram 303) were heat tolerant genotypes while 8 genotypes (Jamuni, F 2004, Allahabad Local, Black, U. P Local, YOUDA, U. P B2425 and HD 2967) have been observed high temperature susceptible. While WH-1105 and PBW-154 have been observed high yielding genotypes under heat stress environment. Therefore, genotypes WH-1105 and PBW-154 were acknowledged as a suitable genotype for late sown trial. Further, these two genotypes could be utilize in breeding programme to develope heat tolerant varieties of wheat.

Physiology of carrot seeds treated with red seaweed biostimulant and exposed to different temperatures and salinity

Carrot (Daucus carota L.) is a horticulture crop species with economic importance. The performance of this crop can be improved with the use of seaweed extracts, which are a sustainable alternative to chemical fertilizers and biostimulants. Few studies have been performed aiming to determine the effects of algae-based biostimulants, especially red algae of the genus Solieria, on seed treatment and physiology. This study aimed to evaluate the effect of seed treatment with Solieria filiformis algal biostimulant for improving the physiological potential of carrot seeds of Brasilia Irecê and Nantes cultivars. Carrot seeds of both cultivars were treated with 0, 1, 2, and 4 mL L−1 of the biostimulant, after which they were subjected to germination and seedling growth evaluation at different temperatures and salinities. The results were analyzed by analysis of variance, Tukey’s test, and regression. It was observed that 30 °C temperature reduced carrot germination and seedling growth for the cultivars. No dose of red algae biostimulant improved germination of carrot seeds under both adequate and stress conditions of temperature and salinity for the cultivars analyzed.

Genome-Wide Identification, Evolution, and Expression Analysis of the MAPK Gene Family in Rosaceae Plants

Mitogen-activated protein kinases (MAPKs) are crucial regulators in coping with abiotic and biotic stresses, including drought, salinity, fungi, and pathogens. However, little is known about the characteristics, evolution process, and functional divergence of the MAPK gene family in Rosaceae plants. A total of 97 MAPK members were identified in six Rosaceae species, including 12 genes in Fragaria vesca, 22 genes in Malus domestica, 23 genes in Pyrus bretschneideri, 12 genes in Prunus mume, 14 genes in Prunus persica, and 14 genes in Rosa chinensis. All MAPK members of six Rosaceae plants were categorized into four clusters by the phylogenetic relationship analysis. Collinearity analysis discovered that both segmental duplication and tandem duplication contributed to the expansion of MAPK family genes in Rosaceae plants. And the analysis of motifs and gene structures indicated that the evolution of the MAPK gene family was highly conserved among phylogenetic clusters in Rosaceae species. In addition, the dN/dS rates of MAPK paralogous gene pairs were below one, suggesting the MAPK gene family in Rosaceae was driven by purifying selective pressure. Furthermore, functional divergence analysis discovered that 14 amino acid residues were detected as potentially key sites for functional divergence of MAPK family genes between different cluster pairs, specifically Type I functional divergence. The analysis of functional distance indicated that cluster C retained more of the original functional features, while cluster B exhibited functional specialization. Moreover, the expression profiles revealed that PmMAPK8, PmMAPK9, and PmMAPK10 were both highly expressed under drought stress and low temperature conditions. This study aims to comprehensively analyze the evolutionary process and functional analyses of the MAPK gene family in Rosaceae plants, which will lay the foundation for future studies into MAPK genes of Rosaceae in response to drought and cold stress.

Infliximab aggregates produced in severe and mild elevated temperature stress conditions induce an extended specific CD4 T-cell response

Aggregation has been widely described as a factor contributing to therapeutic antibody immunogenicity. Although production of high-affinity anti-drug antibodies depends on the activation of CD4 T lymphocytes, little is known about the T-cell response induced by antibody aggregates, especially for aggregates produced in mild conditions resulting from minor handling errors of vials. Large insoluble infliximab (IFX) aggregates produced in severe elevated temperature stress conditions have been previously shown to induce human monocyte-derived dendritic cell (moDC) maturation. We here showed that large IFX aggregates recruit in vitro a significantly higher number of CD4 T-cells compared to native IFX. Moreover, a larger array of T-cell epitopes encompassing the entire variable regions was evidenced compared to the native antibody. We then compared the responses of moDCs to different types of aggregates generated by submitting IFX to mild conditions of various times of incubation at an elevated temperature. Decreasing stress duration reduced aggregate size and quantity, and subsequently altered moDC activation. Of importance, IFX aggregates generated in mild conditions and not altering moDC phenotype generated an in vitro T-cell response with a higher frequency of CD4 T cells compared to native IFX. Moreover, cross-reactivity studies of aggregate-specific T cells showed that some T cells could recognize both native and aggregated IFX, while others responded only to IFX aggregates. Taken together, our results suggest that aggregation of antibodies in mild elevated temperature stress conditions is sufficient to alter moDC phenotype in a dose-dependent manner and to increase T-cell response.