Transcriptome-wide identification of the Hsp70 gene family in Pugionium cornutum and functional analysis of PcHsp70-5 under drought stress.

MAIN CONCLUSION: The PcHsp70-5 enhances drought stress tolerance in transgenic Arabidopsis thaliana by upregulating stress tolerance genes and antioxidant enzyme activities. Heat shock proteins (HSPs) constitute a class of evolutionarily conserved proteins synthesized by organisms in response to various adverse environmental stimuli such as elevated temperatures, drought, hormonal fluctuations, high salt concentrations, and mechanical stress. However, research on HSPs has predominantly focused on model plants and crops, whereas their functions in desert plants have not been well investigated. This study analyzed the transcriptome of Pugionium cornutum and identified the complete ORFs of 25 genes of the PcHsp70 family genes. Their expression levels under drought stress were investigated using existing RNA-seq data. PcHsp70-5 genes exhibited high expression levels in both roots and leaves under drought stress. Consequently, the PcHsp70-5 genes were cloned and transformed into Arabidopsis thaliana for further analysis of their roles in drought stress response. Real-time fluorescence quantitative PCR (qRT-PCR) analysis demonstrated that both, drought stress and ABA, induced PcHsp70-5 expression. Under drought conditions, transgenic Arabidopsis plants exhibited markedly enhanced growth compared to wild-type plants, as evidenced by improved survival rates, root length, fresh weight, chlorophyll content, and reduced levels of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in leaves, indicating that PcHsp70-5 overexpression mitigated growth inhibition and oxidative damage induced by drought stress. Subsequent research revealed that PcHsp70-5 overexpression significantly augmented the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and increased the proline content in transgenic Arabidopsis under drought conditions, alongside a significant increase in the expression levels of genes related to stress tolerance. This suggests that PcHsp70-5 enhances drought stress tolerance in transgenic Arabidopsis by upregulating stress tolerance genes and antioxidant enzyme activities.

Association of single nucleotide polymorphisms in heat stress protein 70 (HSP70) and 90 (HSP90) with the susceptibility of Pakistani sheep breeds to hemoparasitic infections.

The present study was designed to report the genotypic and allelic frequency of single nucleotide polymorphism (SNP) at 222 G > A in HSP70 and at ex6-7390T22G in the HSP90 gene of 204 sheep (Baluchi = 11, Kajli = 29, Latti = 06 and Mundri = 158) enrolled from District Rajanpur in Punjab and to report the susceptibility of these sheep to the blood-borne parasitic infection. The tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) approach revealed a significant variation (p < 0.001) in the genotype frequency of four enrolled sheep breeds at SNP 222 G > A in the HSP70 gene while the allelic frequency remained unaffected (p = 0.08). In all sheep breeds, GG (wild) genotype was most common. T-ARMS-PCR analysis revealed a similar trend for ex6-7390T22G in the HSP90 gene and it was observed that sheep had significantly higher wild-type (GG) (p < 0.05) at the studied SNPs. Studied epidemiological factors (sex and sampling sites) were not found associated with both SNPs. Chi-square test revealed that no specific genotype and allelic frequency at 222 G > A in HSP70 and at ex6-7390T22G in the HSP90 gene of the enrolled sheep breed was associated with the susceptibility to blood-borne parasitic infection (p > 0.05). In conclusion, we are reporting that Pakistan is blessed to have majority of sheep, from all breeds, having wild genotype at analyzed SNPs in heat stress genes. We highly recommend the genotypic screening of sheep before their selection as breeders to reduce the possibility of having sheep with polymorphic genotypes at 222 G > A in HSP70 and at 7390T22G in HSP90 genes that will improve the profitability and sustainability of animal production systems in Pakistan.

Genome-wide analysis of HSP70 gene superfamily in Pyropia yezoensis (Bangiales, Rhodophyta): identification, characterization and expression profiles in response to dehydration stress.

BACKGROUND: Heat shock proteins (HSPs) perform a fundamental role in protecting plants against abiotic stresses. Individual family members have been analyzed in previous studies, but there has not yet been a comprehensive analysis of the HSP70 gene family in Pyropia yezoensis. RESULTS: We investigated 15 putative HSP70 genes in Py. yezoensis. These genes were classified into two sub-families, denoted as DnaK and Hsp110. In each sub-family, there was relative conservation of the gene structure and motif. Synteny-based analysis indicated that seven and three PyyHSP70 genes were orthologous to HSP70 genes in Pyropia haitanensis and Porphyra umbilicalis, respectively. Most PyyHSP70s showed up-regulated expression under different degrees of dehydration stress. PyyHSP70-1 and PyyHSP70-3 were expressed in higher degrees compared with other PyyHSP70s in dehydration treatments, and then expression degrees somewhat decreased in rehydration treatment. Subcellular localization showed PyyHSP70-1-GFP and PyyHSP70-3-GFP were in the cytoplasm and nucleus/cytoplasm, respectively. Similar expression patterns of paired orthologs in Py. yezoensis and Py. haitanensis suggest important roles for HSP70s in intertidal environmental adaptation during evolution. CONCLUSIONS: These findings provide insight into the evolution and modification of the PyyHSP70 gene family and will help to determine the functions of the HSP70 genes in Py. yezoensis growth and development.

In Silico Analysis of HSP70 Gene Family in Bovine Genome.

Heat shock proteins (HSPs), members of molecular chaperones families fulfill essential roles under normal conditions and provide protection and adaptation during and after stress. Among different HSPs, HSP70 kDa family of proteins is most abundant and well-studied in human and mouse but has not yet been characterized in bovines. In silico analysis was performed to characterize members of HSP70 gene family in bovine genome and a total of 17 genes of bovine HSP70 gene family were identified. The members of HSP70 family were distributed over 12 chromosomes with gene size ranging from 1911 (HSPA2) to 54,017 bp (HSPA4). Five genes were intronless, while rest of 12 genes were multiexonic. Phylogenetic analysis of HSP70 gene family distinguished them into eight major evolutionary groups wherein members of group 1 were most divergent and quite dissimilar than from rest of the HSP70 sequences. Domain structure of all bovine HSP70 genes was conserved and three signature patterns HSP70_1, HSP70_2, and HSP70_3 were identified. HSPA8, HSP9, and HSPA1A showed comparatively higher expression in majority of tissues. Like humans, bovine HSP70 family was characterized by remarkable evolutionary diversity. The analysis also suggested resemblance of bovine HSP70 family to that of human compared to mouse. Overall, the study indicates the presence of diversity for structure, function, localization, and expression in the bovine HSP70 family chaperons which could form the basis to understand thermotolerance/adaptive changes in the bovines.

The pattern of HSP70 gene expression, flight activity and temperature in Apis mellifera meda colonies.

Honey bees produce heat shock proteins (HSPs) following biotic and abiotic stressors to protect cells from damage. In the current study, the pattern of HSP70 gene transcription, foraging, and temperature inside and outside the hive and their association in Apis mellifera meda colonies during Ziziphus blooming period were investigated. Therefore, the number of bees entering the hive, temperature inside and outside the hive were recorded in six colonies at different times of the day. Entering and exiting worker bees were sampled in the front of three hives at three times during the day, morning, midday, and afternoon to evaluate HSP70 gene expression by real-time PCR. The results showed that the flight behavior was influenced by the inside and outside hive temperatures, which was lower and higher in the midday and at the end of the day, respectively. The peak amount of HSP70 gene transcription at the midday was associated with the lowest bee flight activity and highest inside and outside the hive temperatures. In addition, the peak of flight activity was associated with intermediate levels of HSP70 gene expression in the afternoon.

Biological Responses to Cadmium Stress in Liverwort Conocephalum conicum (Marchantiales).

Oxidative damage (production and localization of reactive oxygen species) and related response mechanisms (activity of antioxidant enzymes), and induction of Heat Shock Protein 70 expression, have been studied in the toxi-tolerant liverwort Conocephalum conicum (Marchantiales) in response to cadmium stress using two concentrations (36 and 360 µM CdCl2). Cadmium dose-dependent production of reactive oxygen species (ROS) and related activity of antioxidant enzymes was observed. The expression level of heat shock protein (Hsp)70, instead, was higher at 36 µM CdCl2 in comparison with the value obtained after exposure to 360 µM CdCl2, suggesting a possible inhibition of the expression of this stress gene at higher cadmium exposure doses. Biological responses were related to cadmium bioaccumulation. Since C. conicum was able to respond to cadmium stress by modifying biological parameters, we discuss the data considering the possibility of using these biological changes as biomarkers of cadmium pollution.

Molecular cloning, expression HSP70 and its response to bacterial challenge and heat stress in Microptenus salmoides.

The gene encoding HSP70 was isolated from Microptenus salmoides by homologous cloning and rapid amplification of cDNA ends (RACE). The HSP70 transcripts were 2116 bp long and contained 1953 open reading frames encoding proteins of 650 amino acids with a molecular mass of 71.2 kDa and theoretical isoelectric point of 5.22. The qRT-PCR analysis showed that the HSP70 gene was differentially expressed in various tissues under normal conditions, and the highest HSP70 level was observed in the spleen and the lowest levels in the muscle and heart. The clear time-dependent expression level of HSP70 was observed after bacterial challenge and heat stress. A significant increase in HSP70 expression level was detected and reached a maximum at 3 h and 6 h in liver, spleens and gill tissues after Aeromonas hydrophila infection and heat stress, respectively (P < 0.05). As time progressed, the expression of HSP70 transcript was downregulated and mostly dropped back to the original level at 48 h. The concentration of cortisol, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) increased as the time of stress progressed, with the highest level found on 3 h and later declined rapidly and reached to the control levels at the 48 h. Those results suggested that HSP70 was involved in the immune response to bacterial challenge and heat stress. The cloning and expression analysis of the HSP70 provide theoretical basis to further study the mechanism of anti-adverseness in Microptenus salmoides.

Distinct genetic profiles of Leishmania (Viannia) braziliensis associate with clinical variations in cutaneous-leishmaniasis patients from an endemic area in Brazil.

American tegumentary leishmaniasis (ATL) samples obtained from the lesions of patients with typical (n = 25, 29%), atypical (n = 60, 69%) or both (n = 2%) clinical manifestations were analysed by multilocus enzyme electrophoresis, hsp70 restriction-fragment length polymorphism (PCR-RFLP), hsp70 sequencing and phylogenetics methods. The hsp70 PCR-RFLP analysis revealed two different profiles whose the most samples differed from those expected for Leishmania braziliensis and the other Leishmania species tested: of 39 samples evaluated, two (5%) had a restriction profile corresponding to L. braziliensis, and 37 (95%) had a restriction profile corresponding to a variant pattern. A 1300-bp hsp70 gene fragment was sequenced to aid in parasite identification and a phylogenetic analysis was performed including 26 consensus sequences from the ATL patient's samples and comparing to other Leishmania and trypanosomatids species. The dendrogram allowed to observe a potential population structure of L. braziliensis complex in the studied region, emphasizing that the majority of clinical samples presented a variant genetic profile. Of interest, the L. braziliensis diversity was associated with different clinical manifestations whose parasites with hsp70 variant profile were associated with atypical lesions. The results may be helpful to improve the diagnosis, treatment and control measures of the ATL in endemic areas.

[Study of heat shock protein gene 70 (HSP70) expression and its polymorphous marker (+ 1267A> G) in women under exposure to a long-term stress.]

There was studied the HSP70 gene and its polymorphic marker + 1267A> G (rs754888705) in the HSP70 gene expression in32 women under long-term stress, which was caused by inpatient treatment of their oncologic child. A control group included 25 women without long-term stress factors. There was studied a correlation between the expression level of the HSP70 gene in women and their high blood pressure (BP) episodes. The average duration of stress in the group of children's mothers was 7.3 (2.5-11.5) months. Anxiety level according to HADS in the main group was significantly increased and amounted to 8.7 (7-10) points, in the control group 5 (2-7) points. Depression level in the main group was significantly higher and amounted to 7.7 (7-9) points, in the control group 3.3 (1-5) points. Comparison of the HSP70 gene expression level in blood of young women of the main study group with expression level of this gene in the control group showed a statistically significant predominance of HSP70 gene expression in the main group. The episodes of high BP in women who were under stress conditions did not influence the expression level of the HSP70 gene. Analysis of the alleles of the polymorphic marker frequency in the gene HSP70-2 1267A> G (rs754888705) in the main and control groups showed a statistically significant predominance of allele A in the group of mothers under stress, and predominance of the allele G in the control group. Comparative analysis of the polymorphic marker genotype frequencies in the gene HSP70-2 1267A> G (rs754888705) showed that the AA genotype is significantly more frequent in mothers with episodes of BP increase compared to women without it. An increased level of the HSP70 gene expression in peripheral blood leukocytes in mothers with a life-threatening disease of their child indicates the damaging effect of long-lasting psychoemotional stress at the cellular level and activation of the protective reaction mediated by HSP70. Studying of the genotype AA characteristics of the polymorphic marker A1267G (rs754888705) of the HSP70-2 gene in women under long-lasting stress will let us evaluate the role of genetic factors in hypertensive reactions development.

Endocrine-related genes are altered by antibacterial agent triclosan in Chironomus riparius aquatic larvae.

Triclosan (TCS) is an antibacterial agent widely used in personal care and consumer products and commonly detected in aquatic ecosystems. In the present study, the effects of TCS on endocrine-related genes of Chironomus riparius aquatic larvae, a reference organism in aquatic toxicology, were evaluated. Twenty-four-hour in vivo exposures at 10µg/L, 100µg/L, and 1000µg/L TCS revealed that this xenobiotic was able to alter the transcriptional activity of ecdysone receptor gene (EcR), the ultraspiracle gene (usp), the estrogen-related receptor gene (ERR), and the E74 early ecdysone-inducible gene, as measured by real-time RT-PCR. Moreover, the hsp70 gene, a heat shock protein gene, was upregulated after exposure to TCS. The results of the present work provide the first evidence of the potential disruptive effects of TCS in endocrine-related genes suggesting a mode of action that mimics ecdysteroid hormones in insects.

Genetic polymorphisms of hspa1b and hspa1l in infertile men.

OBJECTIVE: To investigate the relationship between the HSP 70 genepolymorphism and primary infertility in males with normal sperm-parameters. METHODS: The case-control study was conducted in Sanliurfa, Turkey, from September 2010 to August 2011and comprised infertile males as cases and healthy fertile controls. Deoxyribonucleicacid was isolated from the blood of both groups, and polymorphisms of the HSPA1B gene (NM_005346.4, GI: 3304):c.1059G>A, (PstI G>A; dbSNP: rs1061581G>A) and HSPA1L gene (NM_005527.3, GI: 3305) c.1478C>T (NcoIC>T, dbSNP: rs2227956) were analysed with polymerase chain reaction-restriction fragment length polymorphism technique. SPSS version 11.5 was used for statistical analysis. RESULTS: Of the 140males in the study, 68(28.5%) were infertile cases and 72(51.4%) fertile controls. There was no statistically significant difference between GA (heterozygous) and AA (homozygous, polymorphic) genotypes of the c.1059G>A polymorphic point of the HSPA1B gene or between the A allele of the cases and controls (p>0.05). There was no statistically significant difference between the CT (heterozygote) and TT (homozygous, polymorphic) genotypes of the c.1478C>T polymorphic area of the HSPA1Lgene or between the T alleles of the cases and the controls (p>0.05). CONCLUSIONS: Infertility in males with normal sperm parameters was not significantly associated with HSPA1B:c.1059G>A and HSPA1L:c.1478C>T gene polymorphisms. Further prospective studies with larger sample sizes and different gene groups are required to clarify the issue.

In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7)M, 10(-5)M, and 10(-3)M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MTT assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmium in all assays performed. A significant reduction in the cells׳ ability to uptake neutral red and viability by MTT assay was recorded, indicating that both cadmium and copper could change the membrane permeability, inducing cellular apoptosis when the concentration of metals reached 10(-3)M. The apoptotic effect may also explain the high level of cytotoxicity found when the leukocytes were exposed to higher concentration of metals. These results demonstrated that toxic effect of copper and cadmium affect on the mechanisms of cell-mediated immunity reducing the immune defences of the organism.

[Identification and expression analysis of the HSP70 gene family under abiotic stresses in Litchi chinensis].

HSP70 protein, as an important member of the heat shock protein (HSP) family, plays an important role in plant growth, development, and response to biotic and abiotic stresses. In order to explore the role of HSP70 gene family members in Litchi chinensis under low temperature, high temperature, drought, and salt stress, bioinformatics methods were used to identify the HSP70 gene family members within the entire L. chinensis genome. The expression of these genes under various abiotic stresses was then detected using quantitative real-time PCR (qRT-PCR). The results showed that the LcHSP70 gene family consisted of 18 members, which were unevenly distributed across ten L. chinensis chromosomes. The LcHSP70 protein contained 479-851 amino acids, with isoelectric points ranging from 5.07 to 6.95, and molecular weights from 52.44 kDa to 94.07 kDa. The predicted subcellular localization showed that LcHSP70 protein was present in the nucleus, cytoplasm, endoplasmic reticulum, mitochondria, and chloroplast. Phylogenetic analysis divided the LcHSP70 proteins into five subgroups, namely Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅵ. The promoter regions of the LcHSP70 genes contained various cis-acting elements related to plant growth, development, hormone response, and stress response. Moreover, the expression of LcHSP70 genes displayed distint tissue-specific expression level, categorized into universal expression and specific expression. From the selected 6 LcHSP70 genes (i.e., LcHSP70-1, LcHSP70-5, LcHSP70-10, LcHSP70-14, LcHSP70-16, and LcHSP70-18), their relative expression levels were assessed under different abiotic stresses using qRT-PCR. The results indicated that the gene family members exhibited diverse responses to low temperature, high temperature, drought, and salt stress, with significant variations in their expression levels across different time periods. These results provide a foundation for further exploration of the function of the LcHSP70 gene family.

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.

Usefulness of Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry in the Characterization of Leishmania Strains Causing Tegumentary Leishmaniasis in Bolivia versus hsp70 Gene Sequencing.

Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is a proteomic technique with proven efficiency in the identification of microorganisms, such as bacteria, fungi, and parasites. The present study aimed to evaluate the usefulness of MALDI-TOF MS for the characterization of Leishmania species circulating in Bolivia using hsp70 gene sequencing as a reference technique. 55 Leishmania strains that were isolated from patients with tegumentary leishmaniasis were analyzed. MALDI-TOF MS identified two species of the L. braziliensis complex (L. braziliensis, n = 26; L. braziliensis outlier, n = 18), one species of the L. guyanensis complex (L. guyanensis, n = 1), one species of the L. lainsoni complex (L. lainsoni, n = 2), and two species of the L. mexicana complex (L. amazonensis, n = 5; and L. garnhami, n = 3). All of the strains were correctly identified at the subgenus, genus, and complex level, but 10 of them (18%) were misidentified as other species within the same complex by the hsp70 gene sequencing, with 7 of these corresponding to possible hybrids. Thus, one L. braziliensis corresponded to L. peruviana, two L. braziliensis corresponded to L. braziliensis/L. peruviana possible hybrids, two L. amazonensis corresponded to L. mexicana, and three L. garnhami and two L. amazonensis corresponded to L. mexicana/L. amazonensis possible hybrids. Accordingly, MALDI-TOF MS could be used as an alternative to molecular techniques for the identification of Leishmania spp., as it is low cost, simple to apply, and able to quickly produce results. In Bolivia, its application would allow for the improvement of the management of patient follow-ups, the updating of the epidemiological data of the Leishmania species, and a contribution to the control of tegumentary leishmaniasis. IMPORTANCE The objective of the study was to evaluate the usefulness of MALDI-TOF MS for the characterization of Leishmania species circulating in Bolivia, in comparison with the sequencing of the hsp70 gene. In our study, all of the isolates could be identified, and no misidentifications were observed at the complex level. Although the equipment implies a high initial investment in our context, MALDI-TOF MS can be used in different areas of microbiology and significantly reduces the cost of testing. Once the parasite culture is obtained, the technique quickly yields information by accessing a free database that is available online. This would allow for the improvement of the management of patients and follow-ups, the updating of the epidemiological data of the species, and a contribution to the control of tegumentary leishmaniasis in Bolivia. Likewise, it can be used to determine a specific treatment to be given, according to the causal species of Leishmania, when there are protocols in this regard in the area.

Genome-wide analysis of Hsp40 and Hsp70 gene family in four cotton species provides insights into their involvement in response to Verticillium dahliae and abiotic stress.

Introduction: Cotton is an important economic crop to provide natural fibers as raw materials to textile industry, and is significantly affected by biotic and abiotic stress during the whole growth stage, in which Verticillium wilt (VW) caused by Verticillium dahliae is one of the most destructive disease to lead to a significant yield reduction. Heat shock proteins (Hsps) are important molecular chaperones, and play crucial roles in plant growth, development, resistance to biotic and abiotic stress. Hsp40 and Hsp70 are two key Hsps in cell chaperone network, however, the function and regulatory mechanism of Hsp40 and Hsp70 members in VW resistance and abiotic stress in cotton are largely unknown. Methods and Results: Herein, a systematic and comprehensive analysis of Hsp40s and Hsp70s in four cotton species of Gossypium arboretum, G. raimondii, G. hirsutum, and G. barbadense were performed. A total of 291 Hsp40s and 171 Hsp70s identified in four Gossypium species. Sequence analysis revealed that all Hsp40 proteins contained J domain that provides the binding sites to Hsp70. Protein-protein interaction prediction analysis displayed that GhHsp40-55 might interact with GhHsp70-2 and GhHsp70-13, suggesting their potential function as protein complex. Promoter cis-acting element analysis demonstrated that multiple cis-elements related to disease and stress response consists in GhHsp40 and GhHsp70 promoters. Further expression analysis showed that eight GhHsp40s (Hsp40-2,4,8,11,20,23,53,55) and seven GhHsp70s (Hsp70-2,3,6,8,13,19,22) were up-regulated after V. dahliae infection. In addition, five GhHsp40s (Hsp40-2,8,11,53,55) and four GhHsp70s (Hsp70-3,6,8,13) were up-regulated after salt treatment, six GhHsp40s (Hsp40-4,11,20,23) and three GhHsp70s (Hsp70-2,8,19) were up-regulated after drought treatment, four GhHsp40s (Hsp40-2,11,20,23) and four GhHsp70s (Hsp70-3,6,19,22) were up-regulated after temperature treatment, suggesting these Hsps have possible important function in the process of abiotic stress response. Discussion: Our results lay a foundation for understanding the function of Hsp40 and Hsp70 in the resistance against V. dahliae and abiotic stress, and elucidating the regulatory mechanism of the protein complex, evolution and molecular mechanism under stress.

Transcriptome profiling reveals the strategy of thermal tolerance enhancement caused by heat-hardening in Mytilus coruscus.

The thick-shell mussel Mytilus coruscus serves as a common sessile intertidal species and holds economic significance as an aquatic organism. M. coruscus often endure higher temperatures than their ideal range during consecutive low tides in the spring. This exposure to elevated temperatures provides them with a thermal tolerance boost, enabling them to adapt to high-temperature events caused by extreme low tides and adverse weather conditions. This phenomenon is referred to as heat-hardening. Some related studies showed the phenomenon of heat-hardening in sessile intertidal species but not reported at the mechanism level based on transcriptome so far. In this study, physiological experiments, gene family identification and transcriptome sequencing were performed to confirm the thermotolerance enhancement based on heat-hardening and explore the mechanism in M. coruscus. A total of 2935 DEGs were identified and the results of the KEGG enrichment showed that seven heat-hardening relative pathways were enriched, including Toll-like receptor signal pathway, Arachidonic acid metabolism, and others. Then, 24 HSP70 members and 36 CYP2 members, were identified, and the up-regulated members are correlated with increasing thermotolerance. Finally, we concluded that the heat-hardening M. coruscus have a better thermotolerance because of the capability of maintaining the integrity and the phenomenon of vasodilation of the gill under thermal stress. Further, the physiological experiments yielded the same conclusions. Overall, this study confirms the thermotolerance enhancement caused by heat-hardening and reveals the survival strategy in M. coruscus. In addition, the conclusion provides a new reference for studying the intertidal species' heat resistance mechanisms to combat extreme heat events and the strategies for dealing with extreme weather in aquaculture under the global warming trend.

Genetic profiling of HSP70 gene in local Iraqi goats.

Animals display numerous physiological and behavioral responses that reduce the effects of heat stress. Moreover, genetic variance is strongly associated with responses to heat stress, including variants of heat shock proteins (HSPs) that are necessary for thermoregulation and stress resistance. Herein, we performed the molecular profiling of the HSP70 gene, and its polymorphism was demonstrated as a possible factor in the stress tolerance of local Iraqi goats. A number of different mutations were found owing to seven main polymorphisms. Results indicated the occurrence of silent and missense mutations in sequences obtained for Iraqi local goats. Genetic diversity was observed in the HSP70 gene of Iraqi local goats on the basis of phylogenetic-tree analysis as some mutations occurred once whereas others occurred multiple times. The polymorphisms LC616787, LC616788, and LC616791 were combined with the reference gene in the same branch, whereas polymorphisms (LC616785 and LC616786) and (LC616789 and LC616790) met in different branches, respectively. Moreover, all studied proteins had mismatches in their three-dimensional structures. Therefore, the presence of specific genetic differences within the HSP70 gene in Iraqi goats can increase the possibility of selecting animals more suitable to various levels of stress.

Os animais apresentam uma série de respostas fisiológicas e comportamentais que reduzem os efeitos do estresse térmico. Além disso, a variância genética está fortemente associada às respostas ao estresse térmico, incluindo variantes de proteínas de choque térmico (HSPs) que também são necessárias para a termorregulação e resistência ao estresse. O perfil molecular do gene HSP70 foi realizado neste estudo e o polimorfismo desse gene foi demonstrado como um possível fator na tolerância ao estresse de caprinos iraquianos. Várias mutações diferentes foram encontradas devido a sete polimorfismos principais. Os resultados indicam a ocorrência de mutações silenciosas e sem sentido em sequências obtidas para caprinos iraquianos. A diversidade genética pode ser vista no gene HSP70 de cabras locais iraquianas com base na análise da árvore filogenética, já que algumas mutações ocorreram uma vez, enquanto outras ocorreram várias vezes. Os polimorfismos LC616787, LC616788 e LC616791 foram combinados com o gene de referência no mesmo ramo, enquanto os polimorfismos (LC616785 e LC616786) e (LC616789 e LC616790) se encontraram em diferentes ramos, respectivamente. O estudo também revelou que todas as proteínas estudadas tinham incompatibilidade sem suas estruturas tridimensionais. De acordo com nossas descobertas, a presença de diferenças genéticas específicas dentro do gene HSP70 em caprinos iraquianas aumentaria a possibilidade de seleção de animais mais adequados a vários níveis de estresse.

Tegumentary leishmaniasis by Leishmania braziliensis complex in Cochabamba, Bolivia including the presence of L. braziliensis outlier: Tegumentary leishmaniasis in Cochabamba, Bolivia: Tegumentary leishmaniasis in Cochabamba, Bolivia.

Leishmaniasis is caused by protozoans of the Leishmania genus, which includes more than 20 species capable of infecting humans worldwide. In the Americas, the most widespread specie is L. braziliensis, present in 18 countries including Bolivia. The taxonomic position of the L. braziliensis complex has been a subject of controversy, complicated further by the recent identification of a particular subpopulation named L. braziliensis atypical or outlier. The aim of this study was to carry out a systematic analysis of the L. braziliensis complex in Bolivia and to describe the associated clinical characteristics. Forty-one strains were analyzed by sequencing an amplified 1245 bp fragment of the hsp70 gene, which allowed its identification as: 24 (59%) L. braziliensis, 16 (39%) L. braziliensis outlier, and one (2%) L. peruviana. In a dendrogram constructed, L. braziliensis and L. peruviana are grouped in the same cluster, whilst L. braziliensis outlier appears in a separate branch. Sequence alignment allowed the identification of five non-polymorphic nucleotide positions (288, 297, 642, 993, and 1213) that discriminate L. braziliensis and L. peruviana from L. braziliensis outlier. Moreover, nucleotide positions 51 and 561 enable L. peruviana to be discriminated from the other two taxa. A greater diversity was observed in L. braziliensis outlier than in L. braziliensis-L. peruviana. The 41 strains came from 32 patients with tegumentary leishmaniasis, among which 22 patients (69%) presented cutaneous lesions (11 caused by L. braziliensis and 11 by L. braziliensis outlier) and 10 patients (31%) mucocutaneous lesions (eight caused by L. braziliensis, one by L. braziliensis outlier, and one by L. peruviana). Nine patients (28%) simultaneously provided two isolates, each from a separate lesion, and in each case the same genotype was identified in both. Treatment failure was observed in six patients infected with L. braziliensis and one patient with L. peruviana.

Genome-wide identification and phylogenetic relationships of the Hsp70 gene family of Aegilops tauschii, wild emmer wheat (Triticum dicoccoides) and bread wheat (Triticum aestivum).

Heat shock protein 70 (Hsp70) plays an important role in plant development. It is closely related to the physiological process of cell development and the response to abiotic and biological stress. However, the classification and evolution of Hsp70 genes in bread wheat, wild emmer wheat and Aegilops tauschii are still unclear. Therefore, this study conducted a comprehensive bioinformatics analysis of Hsp70 gene in three species. Among these three species, 113, 79 and 36 Hsp70 genes were identified. They are divided into six subfamilies. Group vi-1 is different from Arabidopsis thaliana. It may be the result of early evolutionary segregation. The number of exons in different subfamilies (from 1 to 13) was different, but the distribution patterns of exons / introns in the same subfamily were similar. The results of Hsp70 promoter region analysis showed that the cis-regulatory elements of A. tauschii and wild emmer wheat were different from those of wheat. In addition, CpG island proportion of wild emmer Hsp70 was higher than that of wheat, which may be the molecular basis of heat resistance of wild wheat relative to cultivated wheat. Further comprehensive analysis of chromosome location and repeat events of Hsp70 gene showed that whole-genome duplication and tandem duplication events contributed to the evolution and expansion of Hsp70 gene in wheat. The results of non-synonymous substitution and synonymous substitution analysis showed that Hsp70 genes of three species had undergone purification selection. The expression profile analysis showed that Hsp70 gene was highly expressed in the roots during the vegetative growth period. In addition, TaHsp70 gene was highly expressed under various stress. The identification, classification and evolution of Hsp70 in wheat and its relatives provided a basis for further research on its evolution and its molecular mechanism in response to stress. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02639-5.