Genome-Wide Identification and Characterization of HSP70 Gene Family in Aquilaria sinensis (Lour.) Gilg

The heat shock protein 70 (HSP70) gene family perform a fundamental role in protecting plants against biotic and abiotic stresses. Aquilaria sinensis is a classic stress-induced medicinal plant, producing a valuable dark resin in a wood matrix, known as agarwood, in response to environmental stresses. The HSP70 gene family has been systematic identified in many plants, but there is no comprehensive analysis at the genomic level in A. sinensis. In this study, 15 putative HSP70 genes were identified in A. sinensis through genome-wide bioinformatics analysis. Based on their phylogenetic relationships, the 15 AsHSP70 were grouped into six sub-families that with the conserved motifs and gene structures, and the genes were mapped onto six separate linkage groups. A qRT-PCR analysis showed that the relative expression levels of all the AsHSP70 genes were up-regulated by heat stress. Subcellular localization of all HSP70s was predicted, and three were verified by transiently expressed in Arabidopsis protoplasts. Based on the expression profiles in different tissues and different layers treated with Agar-Wit, we predict AsHSP70 genes are involved in different stages of agarwood formation. The systematic identification and expression analysis of HSP70s gene family imply some of them may play important roles in the formation of agarwood. Our findings not only provide a foundation for further study their biological function in the later research in A. sinensis, but also provides a reference for the analysis of HSPs in other species.

Stress Resistance Heat Shock Protein 70 (HSP70) Analysis in Sorghum (Sorghum bicolor L.) at Genome-Wide Level

Heat shock proteins (HSP70) play an important role in many biological processes. However, as typical in Sorghum bicolor, the systematic identification of the HSP70 gene is very limited, and the role of the Hsp70 gene in the evolution of Sorghum bicolor has not been described systematically a lot. To overcome the gap, Insilco analysis of HSP70 gene family was conducted.The investigation was utilizing the bioinformatics method to analyze the HSP70 gene family and it has been identified that 30 HSP70 genes from the genome sequence of Sorghum bicolor. A comprehensive analysis of these 30 identified genes undertaking the analysis of gene structure, phylogeny, and physicochemical properties, subcellular localization, and promoter region analysis. The gene structure visualization analyses revealed that 22 genes contains both 5’ and 3’ UTRS and one 5’ and one 3’ gene and 6 genes without UTR. The highest number of introns was recorded as 12 and those genes have shown that without in any intron. In the promoter region analysis, ten protein motifs are identified and characterized and 2219 cis-acting elements are identified. Among those, the promoter enhancer elements share the highest number (1411) and light-responsive elements share the next value (335). The physicochemical properties analysis revealed that 23 families have an acidic nature while four families are basic and the rests are neutral. In general, the different analyses performed disclosed their structural organization, subcellular localization, physicochemical properties, cis-acting elements, phylogenetic, and understress conditions. This study provides further information for the functional characterization of HSP70 and helps to understand the mechanisms of abiotic stress tolerance under diverse stress conditions in Sorghum bicolor.

The functional response of the Antarctic bivalve Laternula elliptica to ocean warming and acidification

<p>Marine life is currently under threat from large-scale, long-term changes to the marine environment. Anthropogenic emissions of greenhouse gases, particularly carbon dioxide (CO₂), are causing ongoing change to global marine systems, particularly through ocean warming and acidification. Greenhouse gases in the atmosphere are trapping radiation and heating the entire Earth surface, including the ocean. At the same time, oceanic uptake of CO₂ through absorption by surface waters is altering ocean chemistry, increasing acidity, reducing availability of carbonate ions (CO₃²⁻), and causing increasing dissolution of calcium carbonate (CaCO₃) structures. Because atmospheric CO₂ diffuses more readily into cold water, the Southern Ocean (SO) will experience ocean acidification in a matter of decades. Warming in the SO is also occurring rapidly and represents a comparatively greater increase in temperature than elsewhere. SO marine fauna have evolved in constant, stable, cold conditions, and as a result are stenothermal and particularly at risk from ocean warming and acidification. The large infaunal bivalve Laternula elliptica is a prevalent keystone species found throughout the Antarctic benthos in high numbers, and contributes significantly to biodeposition and bentho-pelagic coupling. This thesis examines how L. elliptica adults are affected over medium-term (5-mo) timescales by SO warming and acidification. Adult L. elliptica collected from Cape Evans in McMurdo Sound, Antarctica, were subjected to combinations of temperatures and pHs predicted for the SO by 2050 and 2100 (Temperatures: -1.4°C (control); -0.5°C; +0.5°C. pHs: pH 8.00 (control); pH 7.85; pH 7.65). L. elliptica were assessed at 5 wk and 5 mo to determine their cellular, metabolic, and whole-organism responses to temperature increase and/or pH decrease. Survival parameters such as final survival percentage, survival curves, and time to 50% survival (LD₅₀) were compared among treatments. L. elliptica survival was severely reduced by warming of only 1-2°C above summer ambient temperatures. Physical and physiological condition indices were calculated to assess health, and show changes in shell and body tissue mass. Physical condition stayed similar amongst all treatments at both time points, while physiological condition decreased significantly at 5 mo with elevated temperature. Oxygen (O₂) consumption was measured as a proxy for standard metabolic rate to show whether animals had acclimatised to conditions. O₂ consumption was significantly negatively correlated with physiological condition, and increased, becoming more variable, with both elevated temperature and lowered pH. This indicated that L. elliptica experienced increased metabolic demand in response to these conditions, and there was a general lack of acclimation to these conditions over time. Overall, pH had no significant effect on survival, metabolic rate, or condition. Heat shock protein 70 (HSP70) gene expression levels were measured to provide a preliminary indication of how the heat shock response of L. elliptica responds to both elevated temperature and reduced pH. Lowered pH appeared to stimulate an up-regulation of HSP70 gene expression at both time points, although this was smaller at 5 mo. L. elliptica did not seem to display a heat shock response at environmentally realistic levels of warming. Overall, warming resulted in lowered survival and condition loss with no sign of acclimation after 5 mo. These responses occurred at smaller degrees of warming than are typically considered lethal for L. elliptica, indicating that successful longer-term maintenance is more thermally limited than short-term survival in this species. While physical (shell) condition was maintained in undersaturated conditions under both elevated temperature and reduced pH, this maintenance occurred alongside increased O₂ demand. Maintaining the aragonitic shell in combination with increased metabolic activity may have contributed to the decline in physiological (body mass) condition observed in L. elliptica. In combination, the results of this experiment indicate that warming of the SO may be more important than ocean acidification to the survival and functioning of adult L. elliptica.</p>

The functional response of the Antarctic bivalve Laternula elliptica to ocean warming and acidification

<p>Marine life is currently under threat from large-scale, long-term changes to the marine environment. Anthropogenic emissions of greenhouse gases, particularly carbon dioxide (CO₂), are causing ongoing change to global marine systems, particularly through ocean warming and acidification. Greenhouse gases in the atmosphere are trapping radiation and heating the entire Earth surface, including the ocean. At the same time, oceanic uptake of CO₂ through absorption by surface waters is altering ocean chemistry, increasing acidity, reducing availability of carbonate ions (CO₃²⁻), and causing increasing dissolution of calcium carbonate (CaCO₃) structures. Because atmospheric CO₂ diffuses more readily into cold water, the Southern Ocean (SO) will experience ocean acidification in a matter of decades. Warming in the SO is also occurring rapidly and represents a comparatively greater increase in temperature than elsewhere. SO marine fauna have evolved in constant, stable, cold conditions, and as a result are stenothermal and particularly at risk from ocean warming and acidification. The large infaunal bivalve Laternula elliptica is a prevalent keystone species found throughout the Antarctic benthos in high numbers, and contributes significantly to biodeposition and bentho-pelagic coupling. This thesis examines how L. elliptica adults are affected over medium-term (5-mo) timescales by SO warming and acidification. Adult L. elliptica collected from Cape Evans in McMurdo Sound, Antarctica, were subjected to combinations of temperatures and pHs predicted for the SO by 2050 and 2100 (Temperatures: -1.4°C (control); -0.5°C; +0.5°C. pHs: pH 8.00 (control); pH 7.85; pH 7.65). L. elliptica were assessed at 5 wk and 5 mo to determine their cellular, metabolic, and whole-organism responses to temperature increase and/or pH decrease. Survival parameters such as final survival percentage, survival curves, and time to 50% survival (LD₅₀) were compared among treatments. L. elliptica survival was severely reduced by warming of only 1-2°C above summer ambient temperatures. Physical and physiological condition indices were calculated to assess health, and show changes in shell and body tissue mass. Physical condition stayed similar amongst all treatments at both time points, while physiological condition decreased significantly at 5 mo with elevated temperature. Oxygen (O₂) consumption was measured as a proxy for standard metabolic rate to show whether animals had acclimatised to conditions. O₂ consumption was significantly negatively correlated with physiological condition, and increased, becoming more variable, with both elevated temperature and lowered pH. This indicated that L. elliptica experienced increased metabolic demand in response to these conditions, and there was a general lack of acclimation to these conditions over time. Overall, pH had no significant effect on survival, metabolic rate, or condition. Heat shock protein 70 (HSP70) gene expression levels were measured to provide a preliminary indication of how the heat shock response of L. elliptica responds to both elevated temperature and reduced pH. Lowered pH appeared to stimulate an up-regulation of HSP70 gene expression at both time points, although this was smaller at 5 mo. L. elliptica did not seem to display a heat shock response at environmentally realistic levels of warming. Overall, warming resulted in lowered survival and condition loss with no sign of acclimation after 5 mo. These responses occurred at smaller degrees of warming than are typically considered lethal for L. elliptica, indicating that successful longer-term maintenance is more thermally limited than short-term survival in this species. While physical (shell) condition was maintained in undersaturated conditions under both elevated temperature and reduced pH, this maintenance occurred alongside increased O₂ demand. Maintaining the aragonitic shell in combination with increased metabolic activity may have contributed to the decline in physiological (body mass) condition observed in L. elliptica. In combination, the results of this experiment indicate that warming of the SO may be more important than ocean acidification to the survival and functioning of adult L. elliptica.</p>

Association of SSCP Polymorphisms of HSP70 Gene with Physiological, Production and Reproduction Performance in Sahiwal and Crossbred Cows

Background: Cellular tolerance to heat stress is mediated by heat shock proteins (HSPs). The HSPs act as molecular chaperones and are transcribed in response to stress. Among different families of these proteins, HSP70 is considered to be related to the development of temperature tolerance. Unraveling polymorphism in heat shock protein genes could be a step towards the identification of genetic markers for selecting heat-tolerant cattle. Methods: The present study was carried out in Sahiwal (n=50) and Crossbred cows (n=50) with the objective to identify polymorphisms in HSP70 gene. Two fragments (295 and 220 bp) of HSP70 gene were subjected to Polymerase Chain Reaction-Single-Strand Conformation Polymorphism (PCR-SSCP) technique. Statistical analysis was performed to study the association of each SSCP genotype on physiological, production and reproduction traits in Sahiwal and crossbred cows using the univariate GLM model of SPSS 25. Result: The PCR-SSCP of 295 bp fragment of HSP70 gene revealed two genotypes AA and AB in Sahiwal cows and two genotypes AA and AC in crossbred cows. The association analysis revealed that genotype AA had higher peak milk yield in Sahiwal cows while the same genotype had higher total lactation milk yield, lower service period and calving interval in crossbred cows. The 220 bp fragment was found to be monomorphic in both Sahiwal and crossbred cows.

Assessment of Responses of Common Carp, Cyprinus carpio, Exposed to Sub-Lethal Concentrations of Lead, by Molecular and Mucosal Biomarkers

Heavy metals are one of the most persistent pollutants in waters, and molecular biomarkers as bioindicators could uncover early warning signals of stress suffered by organisms exposed to these pollutants. This research was conducted to explore the impact of lead contaminant on expression levels of hsp70 gene as well as protein pattern and innate immune indicators of skin mucus in Common carp as a biological model. The fish were exposed to different concentrations of Pb(NO3)2 as a source of lead (0, 7.875, 15.75, and 31.5 mg/L of lead) for 14 days. After the completion of the experiment, the liver and gill tissue plus mucus samples were collected from the fish. Next, the effects of exposure to sublethal doses of lead were investigated on expression levels of hsp70 gene, protein pattern, and innate immune indicators of skin mucus. The results indicated that the expression level of hsp70 gene significantly increased (at 7.5% & 30% LC50) in the liver tissue compared to the control group (P<0.05). The protein pattern of treatment samples was different from that of the control group. Also, in the mucus innate immune parameters, the levels of alkaline phosphatase increased significantly at high concentrations of lead contamination in comparison with the control group (P<0.05). The protein levels first revealed an ascending trend (P<0.05), and then a descending trend (at 30% LC50), though it was not a significant downward trend compared with the control group (P>0.05). The results suggested that these physiological indicators in Cyprinus carpio could be used as molecular and mucosal biomarkers for ecotoxicological studies.

Molecular Characterization of the Coding Region and 5’ UTR of HSP70 Gene in Indian Riverine Buffalo Breeds

Background: HSP70 (Heat Shock Protein 70), plays a crucial role in nascent protein folding; the added challenges due to physiological factors demand stringent role-playing of such chaperones for tropical livestock such as water buffalo (Bubalus bubalis). Therefore to evaluate the variations at nucleotide level in HSP70 that could potentially unravel the molecular basis of thermal adaptation in the riverine buffalo breeds of India, the current study was targeted to sequence the CDS (Coding Sequence) and UTR (Untranslated Region) of the gene in a panel of 16 Indian riverine buffalo breeds. Methods: Blood samples were collected and genomic DNA was isolated followed by PCR standardized for the amplification of different fragments of the HSP70 gene using different sets of primer pairs covering the entire coding region and 5’UTR. Multiple amplicons generated to cover the entire gene were sequenced. Sequences were further analyzed manually for the identification of heterozygous animals to detect the polymorphic nucleotide sites and variation between breeds documented. Result: The HSP70 results suggest, the highly conserved nature of gene in buffalo. The only non-synonymous polymorphic site was found in the Toda buffalo breed (g.SNPC greater than T at position 14), resulting in amino acid change 5M greater than T. A total of 7 polymorphic sites were found in the 5’UTR flanking region. Additionally, two insertion/deletions (INDEL) of 30 and 1 nucleotide length were found in the 5’UTR.

Local adaptation of invasive plant, Synedrella nodiflora, in urban tropical lowland landscape Universitas Indonesia

Synedrella nodiflora is an invasive species originated from tropical America and now has spread throughout Indonesia. We analysed the ability of Synedrella nodiflora from the level of HSP70 gene expression at different heat stress in urban tropical lowland landscape Universitas Indonesia. We used the qPCR to quantify the level of HSP70 gene expression and analysed using Pfaffl model. We found the level of HSP70 gene expression got higher related to elevated temperature from 29oC to 39oC with a range of fold from 123.1 to 1676.9. This ability reflects the adaptive plasticity of Synedrella nodiflora in the course of the invasion process.