Small Heat Shock Protein Gene Research Articles

Mitochondrial small heat shock protein and chilling tolerance in tomato fruit

Our previous report indicated that tomato (Solanum lycopersicum) fruit of two contrasting varieties in the chilling tolerance showed the opposite expression pattern of a mitochondrial small heat shock protein (M-sHSP23.8) gene after chilling stress. Thus, the fruit of the relatively tolerant variety Micro-Tom strongly accumulated M-sHSP23.8 transcripts while the susceptible var. Minitomato fruit did not. To test whether M-sHSP23.8 is involved in tomato fruit protection mechanisms against chilling stress, Minitomato fruit overexpressing M-sHSP23.8 (OE23.8) and knockdown Micro-Tom fruit with reduced levels of M-sHSP23.8 (amiR23.8) were developed. After chilling treatment, most of the amiR23.8 fruit failed to ripen normally, showed wilting and skin wrinkles, partial discoloration, and did not reach full red color. On the contrary, these chilling injury symptoms were significantly diminished in OE23.8 fruit, showing less visible deterioration after chilling. Fruit of OE23.8 and amiR23.8 showed opposite patterns of water loss, electrolyte leakage, and expression of the tomato catalase 1 gene compared to control fruit. Membrane lipidome profile evidenced that amiR23.8 fruit showed differential adjustment of extraplastidic and plastidic lipids and variations in the lipid remodeling compared to control fruit, suggesting alterations in the membrane integrity. The high sensitivity of Micro-Tom amiR23.8 fruit and the better performance of Minitomato OE23.8 fruit to chilling treatment indicate that sHSP23.8 may be crucial in the chilling stress tolerance in tomato fruit.

The Chloroplastic Small Heat Shock Protein Gene KvHSP26 Is Induced by Various Abiotic Stresses in Kosteletzkya virginica.

Small heat shock proteins (sHSPs) are a group of chaperone proteins existed in all organisms. The functions of sHSPs in heat and abiotic stress responses in many glycophyte plants have been studied. However, their possible roles in halophyte plants are still largely known. In this work, a putative sHSP gene KvHSP26 was cloned from K. virginica. Bioinformatics analyses revealed that KvHSP26 encoded a chloroplastic protein with the typical features of sHSPs. Amino acid sequence alignment and phylogenetic analysis demonstrated that KvHSP26 shared 30%-77% homology with other sHSPs from Arabidopsis, cotton, durian, salvia, and soybean. Quantitative real-time PCR (qPCR) assays exhibited that KvHSP26 was constitutively expressed in different tissues such as leaves, stems, and roots, with a relatively higher expression in leaves. Furthermore, expression of KvHSP26 was strongly induced by salt, heat, osmotic stress, and ABA in K. virginica. All these results suggest that KvHSP26 encodes a new sHSP, which is involved in multiple abiotic stress responses in K. virginica, and it has a great potential to be used as a candidate gene for the breeding of plants with improved tolerances to various abiotic stresses.

Paeoniflorin Protects Caenorhabditis elegans from Heat Stress

Paeonia lactiflora Pall., a plant of the genus Paeonia, which includes Radix paeoniae Rubra and R. paeoniae Alba has a long history in the traditional Chinese medicine for the management of a variety of ailments. These multiple therapeutic activities have been associated with paeoniflorin, an active component of the roots of R. paeoniaes. Herein, we have evaluated the effect of paeoniflorin on the survival, exercise capacity, antioxidant enzyme activity, and expression of genes hsp-16.2 and daf-16 in C. elegans, a soil-borne free-living nematode, under heat stress at 35°C. Paeoniflorin increased the ability to exercise, survival rate under heat stress, and in vivo activity of antioxidant enzymes significantly. Furthermore, treatment of transgenic C. elegans nematode (TJ375) expressing a jellyfish green fluorescent protein-tagged inducible small heat-shock protein gene with paeoniflorin led to a significant suppression of gene expression. After 2 h of 35°C heat shock exposure there was an enhanced translocation of daf-16 into the nucleus in paeoniflorin-treated nematodes. Similarly, the results of real-time polymerase chain reaction study showed that paeoniflorin promoted expression of daf-16 mRNA levels and reduced the transcription of HSP-16.2 gene under heat stress. In summary, these results show that paeoniflorin has a protective effect on heat stress in C. elegans.

Characterization of Two Small Heat Shock Protein Genes (Hsp17.4 and Hs20.3) from Sitodiplosis mosellana, and Their Expression Regulation during Diapause.

Simple SummarySmall heat shock proteins (sHsps) play important roles in thermal adaptation of various organisms, and insect diapause. Sitodiplosis mosellana, a key pest of wheat worldwide, undergoes obligatory larval diapause in soil to survive adverse temperature extremes during hot summers and cold winters. The objectives of this study were to characterize two sHsp genes from S. mosellana (SmHsp17.4 and SmHsp20.3), and determine their expression in response to diapause, extreme high/low temperatures, or 20-hydroxyecdysone (20E) treatment. Expression of SmHsp17.4 was down-regulated upon entry into diapause, but up-regulated during the shift to post-diapause quiescence. In contrast, expression of SmHsp20.3 was not affected by entry into diapause, but was pronounced during summer and winter. Furthermore, transcripts of both SmHsps were highly responsive to heat (≥35 °C) and cold (≤−5 °C) during diapause, and topical application of 20E on diapausing larvae also induced SmHsp17.4 in a dose-dependent manner. Notably, the recombinant SmHsp17.4 and SmHsp20.3 exhibited significant molecular chaperone activity. In conclusion, SmHsp17.4 and SmHsp20.3 play essential roles in heat/cold adaptation, and 20E-mediated SmHsp17.4 was also likely involved in diapause termination. Results have improved our understanding of the molecular mechanism underlying diapause and related stress tolerance in S. mosellana.Sitodiplosis mosellana, a periodic but devastating wheat pest that escapes temperature extremes in summer and winter by undergoing obligatory diapause. To determine the roles of small heat shock proteins (sHsps) in diapause of S. mosellana, we characterized two sHsp genes, SmHsp17.4 and SmHsp20.3, from this species. Both SmHsps contained the conserved α-crystallin domain and the carboxy-terminal I/VXI/V motif of the sHsp family. SmHsp17.4 had one intron while SmHsp20.3 had none. Quantitative PCR revealed that SmHsp17.4 expression decreased after diapause initiation, but substantially increased during transition to post-diapause quiescence. In contrast, SmHsp20.3 expression was not affected by entry of diapause, but was clearly up-regulated during summer and winter. Short-term more severe heat-stress (≥35 °C) of over-summering larvae or cold-stress (≤−5 °C) of over-wintering larvae could stimulate higher expression of both genes, and SmHsp17.4 was more responsive to cold stress while SmHsp20.3 was more sensitive to heat stress. Notably, transcription of SmHsp17.4, but not SmHsp20.3, in diapausing larvae was inducible by 20-hydroxyecdysone (20E). Recombinant SmHsp17.4 and SmHsp20.3 proteins also displayed significant chaperone functionality. These findings suggest that both SmHsps play key roles in stress tolerance during diapause; and 20E-regulated SmHsp17.4 was also likely involved in diapause termination.

Heterologously Expressing a Wheat CI Small Heat Shock Protein Gene Enhances the Salinity Tolerance of Arabidopsis thaliana

The productivity of wheat, like that of most crop species, is adversely affected by soil salinity, so the identification of genes contributing to the salinity tolerance is an important priority for crop improvement. This study focuses on TaHSP17.6, a gene which encodes a salt and abcisic acid (ABA) inducible class I small molecular heat shock protein. Its transcription level was higher in a salinity tolerant wheat cultivar SR3 than in its less tolerant, but closely related cultivar JN177. The constitutive expression of TaHSP17.6 in Arabidopsis thaliana boosted the plants’ tolerance to salt stress and decreased their sensitivity to exogenously supplied ABA. Compared to wild type plants, the transgenic plants, when challenged with either salinity or ABA, developed a greater number of lateral roots, and their leaves accumulated more proline and have a higher activity of the antioxidant enzyme peroxidase. The transgenic leaves were superior to wild type ones with respect to their ability to maintain their hydration when detached from the plant. The product of TaHSP17.6 is proposed to contribute to the tolerance of osmotic and oxidative stress in an ABA-dependent manner. TaHSP17.6 is proposed as a candidate gene for engineering salinity tolerance in wheat.

Molecular mechanism underlying Pyropia haitanensis PhHsp22-mediated increase in the high-temperature tolerance of Chlamydomonas reinhardtii

Global warming is one of the key limiting factors affecting the cultivation of Pyropia haitanensis which is an economically important macroalgae species grown in southern China. However, the mechanism underlying the high-temperature tolerance of P. haitanensis remains largely unknown. In a previous study, we showed that the expression of the small heat shock protein 22 gene (Hsp22) is upregulated in P. haitanensis in response to high-temperature stress, but the associated regulatory mechanism was not fully elucidated. In this study, a transgenic Chlamydomonas reinhardtii expression system was used to functionally characterize P. haitanensis Hsp22. Our analyses indicated that the C-terminal of PhHsp22 is highly conserved and contains an A-crystal structure domain. A phylogenetic analysis revealed PhHsp22 is not closely related to small heat shock protein genes in other species. Additionally, PhHsp22 expression significantly increased at 3 and 6 h after initiating 33 °C treatment, which improved the survival rate of transgenic C. reinhardtii during the early stage of high-temperature treatment. The further transcriptome analysis revealed that PhHsp22 expression can promote pathways related to energy metabolism, metabolites metabolism, and protein homeostasis in transgenic C. reinhardtii cells exposed to high temperatures. Therefore, PhHsp22 may be crucial for the response of Pyropia species to high-temperature stress. Furthermore, this gene may be useful for breeding new high-temperature algal strains.

Isolation and Characterization of a Rice Mitochondrial Small Heat Shock Protein Gene

To understand the role of small heat shock protein (sHSPs) in rice plant response to various stresses such as the heat and oxidative stresses, a cDNA encoding a 24.1 kDa mitochondrial small HSP (Oshsp24.1) was isolated from rice by rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequence shows very high similarity with other plant small HSPs. DNA gel blot analysis suggests that the rice genome contains more than one copy of Oshsp24.1. High level of expression of Oshsp24.1 transcript was observed in rice seedlings in response to heat, methyl viologen, hydrogen peroxide, ozone, salt and heavy metal stresses. Recombinant OsHSP24.1 protein was produced in E. coli cells for biochemical assay. The protein formed oligomeric complex when incubated with Sulfo-EGS (ethylene glycol bis (succinimidyl succinate)). Our results shows that Oshsp24.1 has an important role in abiotic stress response and have potential for developing stress-tolerant plants.

Knock out of sHSP genes determines some modifications in the probiotic attitude of Lactiplantibacillus plantarum

ObjectiveWe investigated whether the knock out of small heat shock protein (sHSP) genes (hsp1, hsp2 and hsp3) impact on probiotic features of Lactiplantibacillus plantarum WCFS1, aiming to find specific microbial effectors involved in microbe-host interplay.ResultsThe probiotic properties of L. plantarum WCFS1 wild type, hsp1, hsp2 and hsp3 mutant clones were evaluated and compared through in vitro trials. Oro-gastro-intestinal assays pointed to significantly lower survival for hsp1 and hsp2 mutants under stomach-like conditions, and for hsp3 mutant under intestinal stress. Adhesion to human enterocyte-like cells was similar for all clones, though the hsp2 mutant exhibited higher adhesiveness. L. plantarum cells attenuated the transcriptional induction of pro-inflammatory cytokines on lipopolysaccharide-treated human macrophages, with some exception for the hsp1 mutant. Intriguingly, this clone also induced a higher IL10/IL12 ratio, which is assumed to indicate the anti-inflammatory potential of probiotics.ConclusionssHSP genes deletion determined some differences in gut stress resistance, cellular adhesion and immuno-modulation, also implying effects on in vivo interaction with the host. HSP1 might contribute to immunomodulatory mechanisms, though additional experiments are necessary to test this feature.

Differential expression of small heat shock proteins in the brain of broiler embryo; the effects of embryonic thermal manipulation.

Broilers are more vulnerable to high temperatures than mammals due to the feather cover, lack of sweat glands, fast growth and intensive breeding in commercial systems. Thermal stresses affect the function of various organs and change the expression profiles of hundreds of genes in the different tissues of broilers. Thermal manipulation (TM) during embryogenesis can increase heat tolerance in growing broilers. Small heat shock proteins (SHSPs) are a group of HSPs which participate in many cellular functions like response to different stressors. However, their role in the thermotolerance has not been fully elucidated. Ninety fertilized eggs were randomly divided into three groups (30 eggs/group; 10 eggs/replicate). Normal control (NC) eggs were incubated at 37.5°C throughout the incubation period whereas heat stress (HS) and cold stress (CS) groups were kept at 41°C and 33°C from 15 to 17th day of incubation for 3h each day, respectively. On day 20, samples from the cerebrums were harvested for histopathology and mRNA expression analyses of HSPB1, HSPB5, HSPB8, and HSPB9. There were no significant differences in survivability, defected embryos, hatchability, and body weight among treatments. TM had no major deleterious effects on the cerebral tissue except for mild degeneration in the HS group. HSPB1, HSPB5, HSPB8, and HSPB9 were expressed in the presence and absence of TM. All SHSP genes tested were downregulated in response to TM except for HSPB9 which was upregulated in the HS group. The highest change in gene expression due to TM observed for HSPB1. This study presents a broader understanding of mechanisms underlying response to TM in broilers. The results suggest that HSPB1, HSPB5, HSPB8, and HSPB9 are involved in thermotolerance in broilers and SHSPs could be involved in the gene expression profiling of TM. It may propose the use of nutritional supplements in the poultry industry to modulate SHSPs.

Ethylene and RIPENING INHIBITOR Modulate Expression of SlHSP17.7A, B Class I Small Heat Shock Protein Genes During Tomato Fruit Ripening.

Heat shock proteins (HSPs) are ubiquitous and highly conserved in nature. Heat stress upregulates their gene expression and now it is known that they are also developmentally regulated. We have studied regulation of small HSP genes during ripening of tomato fruit. In this study, we identify two small HSP genes, SlHSP17.7A and SlHSP17.7B, localized on tomato Chr.6 and Chr.9, respectively. Each gene encodes proteins constituting 154 amino acids and has characteristic domains as in other sHSP genes. We found that SlHSP17.7A and SlHSP17.7B gene expression is low in the vegetative tissues as compared to that in the fruit. These sHSP genes are characteristically expressed in a fruit-ripening fashion, being upregulated during the ripening transition of mature green to breaker stage. Their expression patterns mirror that of the rate-limiting ethylene biosynthesis gene ACC (1-aminocyclopropane-1-carboxylic acid) synthase, SlACS2, and its regulator SlMADS-RIN. Exogenous application of ethylene to either mature green tomato fruit or tomato leaves suppressed the expression of both the SlHSP17.7A, B genes. Notably and characteristically, a transgenic tomato line silenced for SlACS2 gene and whose fruits produce ~50% less ethylene in vivo, had higher expression of both the sHSP genes at the fruit ripening transition stages [breaker (BR) and BR+3] than the control fruit. Moreover, differential gene expression of SlHSP17.7A versus SlHSP17.7B gene was apparent in the tomato ripening mutants—rin/rin, nor/nor, and Nr/Nr, with the expression of SlHSP17.7A being significantly reduced but that of SlHSP17.7B significantly upregulated as compared to the wild type (WT). These data indicate that ethylene negatively regulates transcriptional abundance of both these sHSPs. Transient overexpression of the ripening regulator SlMADS-RIN in WT and ACS2-AS mature green tomato fruits suppressed the expression of SlHSP17.7A but not that of SlHSP17.7B. Thus, ethylene directly or in tune with SlMADS-RIN regulates the transcript abundance of both these sHSP genes.

Identifications, Characteristics, and Expression Patterns of Small Heat Shock Protein Genes in a Major Mulberry Pest, Glyphodes pyloalis (Lepidoptera: Pyralidae)

Six candidate sHSP genes were identified from the Glyphodes pyloalis transcriptome. All sHSP genes included full-length open reading frames and shared high similarity with the sequences of other lepidopteran species. These sHSP genes encoded 175–191 amino acid residues, and the predicted proteins had a molecular weight from 19.5 to 21.8 kDa. All GpsHSPs were expressed at lower levels at larval stages. All GpsHSPs were expressed at higher levels at diapaused, prepupal, or pupal stages, suggesting that sHSPs may be involved in metamorphosis in G. pyloalis. In addition to the developmental stage, extreme temperatures can induce variations in the expression of sHSPs genes. All GpsHSPs were significantly upregulated in larvae following exposure to heat shock, except GpHSP21.4 which downregulated at 4 h following exposure to the cold shock treatment. Furthermore, Starvation influenced the expression patterns of GpsHSPs as a function of the duration of food deprivation. Four GpsHSPs increased their expression with time of starvation until reaching to the peak level at 6 d of starvation. Finally, parasitism by the endoparasitoid Aulacocentrum confusum He et van Achterberg (Hymenoptera: Braconidae)-induced fluctuations in the expression of all GpsHSPs, and the expression varied with time after parasitization. Our results from this study strongly suggest functional differentiation within the sHSPs subfamily in G. pyloalis. The present study would provide further insight into the roles of sHSPs in G. pyloalis and novel avenues for promoting integrated management of this pest.

A small heat shock protein gene (RcHSP17.8) from Chinese rose confers resistance to various abiotic stresses in transgenic tobacco

RcHSP17.8 isolated from Chinese rose is an induced cytoplasmic class I small heat shock protein, first reported in Chinese rose. To study its physiological function, we inserted the open reading frame of RcHSP17.8 into tobacco and positive transgenic plants were obtained. The transgenic tobacco plants were treated with various stress conditions such as high temperature, drought, osmotic stress, and high salt, and their morphology, water loss rate, electrical conductivity, proline content, and expression patterns of endogenous and exogenous genes were investigated. The results of these physio-biochemical and molecular biological analyses showed that the transgenic tobacco plants exhibited a remarkable resistance phenotype compared with the control (empty vector-transformed tobacco plants). The electrical conductivity was lower in the transgenic tobacco plants compared with the control, while the proline content was substantially increased. Additionally, the transgenic tobacco plants had a reduced water loss rate and higher expression of the endogenous P5CS gene compared with the controls. These results indicated that the RcHSP17.8-transformed tobacco seedlings exhibited remarkable resistance to high temperature and osmotic stresses compared with the control group. This study provides a theoretical basis for genetic engineering research for quality improvement of Chinese rose and other ornamental flowers. RcHSP17.8 was high-expressed in heat-tolerant but not in non-heat-tolerant varieties after heat shock. The tolerance of recombinant strains and transgenic tobacco to abiotic stresses was enhanced.

Small heat shock protein genes are developmentally regulated during stress and non-stress conditions in Blastocladiella emersonii

Small heat shock proteins (sHsps) are molecular chaperones of low molecular weight involved in an early association with misfolded proteins. In response to heat shock, B. emersonii induces the synthesis of a number of proteins. As sHsps are still poorly studied in B. emersonii and in fungi overall, the aim of this work was to carry out a in-depth characterization of sHsps during B. emersonni life cycle, as well as in response to thermal stress. We verified a strong induction of the hsp17 gene in cells exposed to heat shock both in germination and sporulation stages, and that Hsp17 protein levels show the same pattern of variation of its mRNA. Unlike hsp17 and hsp30, hsp16 gene is not significantly induced during heat shock, in germination or sporulation cells. However, at normal temperatures, the hsp16 gene presents high mRNA levels in sporulation cells, whereas the hsp30 gene presents high mRNA levels in germination cells. Interestingly, heat shock mRNA levels for hsp17 and hsp30 genes are 10 times higher in germination cells than in sporulation cells. Thus, our data show that the expression of these sHsp genes is quite distinct, both under normal temperature as during heat shock.

Effects of heat stress on ovarian development and the expression of HSP genes in mice

Heat stress reduces oocyte competence, thereby causing lower fertility in animals. Chronic and acute heat stresses cause extensive morphological damage in animals, but few reports have focused on the effects of chronic and acute heat stresses on ovarian function and heat shock protein (HSP) gene expression during ovarian injury. In this study, we subjected female mice to chronic and acute heat stresses; we then calculated the ovary index, examined ovary microstructure, and measured the expression of multiple HSP family genes. Chronic heat stress reduced whole-body and ovarian growth but had little effect on the ovarian index; acute heat stress did not alter whole-body or ovarian weight. Both chronic and acute heat stresses impaired ovary function by causing the dysfunction of granular cells. Small HSP genes increased rapidly after heat treatment, and members of the HSP40, HSP70, and HSP90 families were co-expressed to function in the regulation of the heat stress response. We suggest that the HSP chaperone machinery may regulate the response to heat stress in the mouse ovary.

Identification of HSF1 Target Genes Involved in Thermal Stress in the Pacific Oyster Crassostrea gigas by ChIP-seq.

The Pacific oyster Crassostrea gigas, a commercially important species inhabiting the intertidal zone, facing enormous temperature fluctuations. Therefore, it is important to identify candidate genes and key regulatory relationships associated with thermal tolerance, which can aid the molecular breeding of oysters. Heat shock transcription factor 1 (HSF1) plays an important role in the thermal stress resistance. However, the regulatory relationship between the expansion of heat shock protein (HSP) HSP 70 and HSF1 is not yet clear in C. gigas. In this study, we analyzed genes regulated by HSF1 in response to heat shock by chromatin immunoprecipitation followed by sequencing (ChIP-seq), determined the expression patterns of target genes by qRT-PCR, and validated the regulatory relationship between one HSP70 and HSF1. We found 916 peaks corresponding to HSF1 binding sites, and these peaks were annotated to the nearest genes. In Gene Ontology analysis, HSF1 target genes were related to signal transduction, energy production, and response to biotic stimulus. Four HSP70 genes, two HSP40 genes, and one small HSP gene exhibited binding to HSF1. One HSP70 with a binding site in the promoter region was validated to be regulated by HSF1 under heat shock. These results provide a basis for future studies aimed at determining the mechanisms underlying thermal tolerance and provide insights into gene regulation in the Pacific oyster.

Detection and molecular characterization of Cryptosporidium species in wild-caught pet spiny-tailed lizards.

Uromastyx is a genus of the herbivorous agamid lizards, also known as spiny-tailed lizards or mastigures, which are found in parts of Africa and the Middle East. Currently, several species of this genus are available in the international pet trade in Japan. In this study, two imported wild-caught spiny-tailed lizards (Arabian blue mastigure, Uromastyx ornata philbyi, and Sudan mastigure, Uromastyx dispar flavifasciata) were diagnosed with a Cryptosporidium (Apicomplexa: Cryptosporidiidae) infection based on the presence of the oocysts in the rectal feces using sucrose flotation and light microscopy examination at a local animal hospital in Tokyo, Japan. One of the lizards had died, and histopathological examination revealed enteritis with the Cryptosporidium parasite. Sequence analyses using the small subunit ribosomal RNA, actin, and 70-kDa heat shock protein genes indicated that the lizards had contracted a novel variant of C. avium that commonly infects avian species.

Genome-wide identification of small heat-shock protein (HSP20) gene family in grape and expression profile during berry development

BackgroundStudies have shown that HSP20 (heat-shock protein 20) genes play important roles in regulating plant growth, development, and stress response. However, the grape HSP20 gene family has not been well studied.ResultsA total of 48 VvHSP20 genes were identified from the grape genome, which were divided into 11 subfamilies (CI, CII, CIII, CV, CVI, CVII, MI, MII, ER, CP and PX/Po) based on a phylogenetic analysis and subcellular localization. Further structural analysis showed that most of the VvHSP20 genes (93.8%) had no intron or only one intron, while genes that clustered together based on a phylogenetic tree had similar motifs and evolutionarily conserved structures. The HSP20s share a conservedα-crystalline domain (ACD) and the different components of the ACD domain suggest the functional diversity of VvHSP20s. In addition, the 48 VvHSP20 genes were distributed on 12 grape chromosomes and the majority of VvHSP20 genes were located at the proximal or distal ends of chromosomes. Chromosome mapping indicated that four groups of VvHSP20 genes were identified as tandem duplication genes. Phytohormone responsive, abiotic and biotic stress-responsive, and plant development-related cis-elements were identified from the cis-regulatory elements analysis of VvHSP20s. The expression profiles of VvHSP20s genes (VvHSP20–1, 11, 14, 17, 18, 19, 20, 24, 25, 28, 31, 39, 42, and 43) were largely similar between RNA-Seq and qRT-PCR analysis after hydrogen peroxide (H2O2) treatment. The results showed that most VvHSP20s were down-regulated by H2O2 treatment during fruit development. VvHSP20s genes were indeed found to be involved in the grape berry development and differences in their transcriptional levels may be the result of functional differentiation during evolution.ConclusionsOur results provide valuable information on the evolutionary relationship of genes in the VvHSP20 family, which is useful for future studies on the functional characteristics of VvHSP20 genes in grape.

Genome-wide identification and transcriptional profiling of small heat shock protein gene family under diverse abiotic stress conditions in Sorghum bicolor (L.)

The small heat shock proteins (sHsps/Hsp20s) are the molecular chaperones that maintain proper folding, trafficking and disaggregation of proteins under diverse abiotic stress conditions. In the present investigation, a genome-wide scan revealed the presence of a total of 47 sHsps in Sorghum bicolor (SbsHsps), distributed across 10 subfamilies, the major subfamily being P (plastid) group with 17 genes. Chromosomes 1 and 3 appear as the hot spot regions for SbsHsps, and majority of them were found acidic, hydrophilic, unstable and intron less. Interestingly, promoter analysis indicated that they are associated with both biotic and abiotic stresses, as well as plant development. Sorghum sHsps exhibited 15 paralogous and 20 orthologous duplications. Expression analysis of 15 genes selected from different subfamilies showed high transcript levels in roots and leaves implying that they are likely to participate in the developmental processes. SbsHsp genes were highly induced by diverse abiotic stresses inferring their critical role in mediating the environmental stress responses. Gene expression data revealed that SbsHsp-02 is a candidate gene expressed in all the tissues under varied stress conditions tested. Our results contribute to the understanding of the complexity of SbsHsp genes and help to analyse them further for functional validation.

Molecular Cloning and Characterization of Small Heat Shock Protein Genes in the Invasive Leaf Miner Fly, Liriomyza trifolii.

Small heat shock proteins (sHSPs) comprise numerous proteins with diverse structure and function. As molecular chaperones, they play essential roles in various biological processes, especially under thermal stresses. In this study, we identified three sHSP-encoding genes, LtHSP19.5, LtHSP20.8 and LtHSP21.7b from Liriomyza trifolii, an important insect pest of ornamental and vegetable crops worldwide. Putative proteins encoded by these genes all contain a conserved α-crystallin domain that is typical of the sHSP family. Their expression patterns during temperature stresses and at different insect development stages were studied by reverse-transcription quantitative PCR (RT-qPCR). In addition, the expression patterns were compared with those of LtHSP21.3 and LtHSP21.7, two previously published sHSPs. When pupae were exposed to temperatures ranging from −20 to 45 °C for 1 h, all LtsHSPs were strongly induced by either heat or cold stresses, but the magnitude was lower under the low temperature range than high temperatures. Developmentally regulated differential expression was also detected, with pupae and prepupae featuring the highest expression of sHSPs. Results suggest that LtsHSPs play a role in the development of the invasive leaf miner fly and may facilitate insect adaptation to climate change.

Identification and Expression Analysis of Four Small Heat Shock Protein Genes in Cigarette Beetle, Lasioderma serricorne (Fabricius).

Small heat shock proteins (sHsps) are molecular chaperones that play crucial roles in the stress adaption of insects. In this study, we identified and characterized four sHsp genes (LsHsp19.4, 20.2, 20.3, and 22.2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The four cDNAs encoded proteins of 169, 180, 181, and 194 amino acids with molecular weights of 19.4, 20.2, 20.3, and 22.2 kDa, respectively. The four LsHsp sequences possessed a typical sHsp domain structure. Quantitative real-time PCR analyses revealed that LsHsp19.4 and 20.3 transcripts were most abundant in pupae, whereas the transcript levels of LsHsp20.2 and 22.2 were highest in adults. Transcripts of three LsHsp genes were highly expressed in the larval fat body, whereas LsHsp20.2 displayed an extremely high expression level in the gut. Expression of the four LsHsp genes was dramatically upregulated in larvae exposed to 20-hydroxyecdysone. The majority of the LsHsp genes were significantly upregulated in response to heat and cold treatments, while LsHsp19.4 was insensitive to cold stress. The four genes were upregulated when challenged by immune triggers (peptidoglycan isolated from Staphylococcus aureus and from Escherichia coli 0111:B4). Exposure to CO2 increased LsHsp20.2 and 20.3 transcript levels, but the LsHsp19.4 transcript level declined. The results suggest that different LsHsp genes play important and distinct regulatory roles in L. serricorne development and in response to diverse stresses.