Small Heat Shock Protein Gene Research Articles

Genome-wide identification and characterization of HSP gene superfamily in whitefly (Bemisia tabaci) and expression profiling analysis under temperature stress.

Heat shock proteins (HSP) are essential molecular chaperones that play important roles in the stress stimulation of insects. Bemisia tabaci, a phloem feeder and invasive species, can cause extensive crop damage through direct feeding and transmission of plant viruses. Here we employed comprehensive genomics approaches to identity HSP superfamily members in the Middle East Asia Minor 1 whitefly genome. In total, we identified 26 Hsp genes, including three Hsp90, 17 Hsp70, one Hsp60 and five sHSP (small heat shock protein) genes. The HSP gene superfamily of whitefly is expanded compared with the other five insects surveyed here. The gene structures among the same families are relatively conserved. Meanwhile, the motif compositions and secondary structures of BtHsp proteins were predicted. In addition, quantitative polymerase chain reaction analysis showed that the expression patterns of BtHsp gene superfamily were diverse across different tissues of whiteflies. Most Hsp genes were induced or repressed by thermal stress (40°C) and cold treatment (4°C) in whitefly. Silencing the expression of BtHsp70-6 significantly decreased the survival rate of whitefly under 45°C. All the results showed the Hsps conferred thermo-tolerance or cold-tolerance to whiteflies that protect them from being affected by detrimental temperature conditions. Our observations highlighted the molecular evolutionary properties and the response mechanism to temperature assaults of Hsp genes in whitefly.

A Class II small heat shock protein OsHsp18.0 plays positive roles in both biotic and abiotic defense responses in rice

Bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most devastating diseases of rice. However, the molecular mechanism underpinning the Xoo resistance of rice is still not fully understood. Here, we report that a class II small heat shock protein gene, OsHsp18.0, whose expression was differentially induced between a resistant and a susceptible variety in response to Xoo infection, plays positive roles in both biotic and abiotic resistance. The molecular chaperone activity of OsHsp18.0 was confirmed by a bacterium-expressed glutathione S-transferase fusion protein. Overexpression of OsHsp18.0 in a susceptible rice variety significantly enhanced its resistance to multiple Xoo strains, whereas silencing of OsHsp18.0 in a resistant variety drastically increased its susceptibility. The enhanced Xoo resistance in OsHsp18.0-overexpressing lines was positively correlated with the sensitized salicylic acid-dependent defense responses. In addition to disease resistance, the OsHsp18.0 overexpressing and silencing lines exhibited enhanced and reduced tolerance, respectively, to heat and salt treatments. The subcellular localization study revealed that the green fluorescent protein-OsHsp18.0 was enriched on the nuclear envelope, suggesting a potential role of OsHsp18.0 in the nucleo-cytoplasmic trafficking. Together, our results reveal that the rice OsHsp18.0 is a positive regulator in both biotic and abiotic defense responses.

Evolutionary Origins of Pax6 Control of Crystallin Genes.

The birth of novel genes, including their cell-specific transcriptional control, is a major source of evolutionary innovation. The lens-preferred proteins, crystallins (vertebrates: α- and β/γ-crystallins), provide a gateway to study eye evolution. Diversity of crystallins was thought to originate from convergent evolution through multiple, independent formation of Pax6/PaxB-binding sites within the promoters of genes able to act as crystallins. Here, we propose that αB-crystallin arose from a duplication of small heat shock protein (Hspb1-like) gene accompanied by Pax6-site and heat shock element (HSE) formation, followed by another duplication to generate the αA-crystallin gene in which HSE was converted into another Pax6-binding site. The founding β/γ-crystallin gene arose from the ancestral Hspb1-like gene promoter inserted into a Ca2+-binding protein coding region, early in the cephalochordate/tunicate lineage. Likewise, an ancestral aldehyde dehydrogenase (Aldh) gene, through multiple gene duplications, expanded into a multigene family, with specific genes expressed in invertebrate lenses (Ω-crystallin/Aldh1a9) and both vertebrate lenses (η-crystallin/Aldh1a7 and Aldh3a1) and corneas (Aldh3a1). Collectively, the present data reconstruct the evolution of diverse crystallin gene families.

Identification and expression analysis of multiple small heat shock protein genes in spruce budworm, Choristoneura fumiferana (L.).

Fifteen small heat shock protein (sHSP) genes were identified from spruce budworm, Choristoneura fumiferana (L.), an important native forest pest in North America. The transcript levels of each CfHSP were measured under non-stress conditions in all life stages from egg to adult and in five different larval tissues. CfHSP transcript levels showed variation during development, with highest levels in adults and lowest in eggs. Most CfHSP transcripts are highly expressed in larval fat body and Malpighian tubules; two CfHSPs display extremely high expression in the head and epidermis. Upon heat stress, nine CfHSP genes are significantly upregulated, increasing by 50- to 2500-fold depending on developmental stage and tissue type. Upon starvation, eight CfHSPs are upregulated or downregulated, whereas six others retain constant expression. These results suggest that CfHSPs have important and multiple roles in spruce budworm development and in response to heat stress and starvation.

Characteristics of six small heat shock protein genes from Bactrocera dorsalis: Diverse expression under conditions of thermal stress and normal growth.

To explore the functions of small heat shock proteins (sHsps) in relation to thermal stress and development in Bactrocera dorsalis (Hendel), one of the most economically important pest species attacking a wide range of fruits and vegetables, six full-length cDNAs of sHsp genes (BdHsp17.7, 18.4, 20.4, 20.6, 21.6 and 23.8) were cloned, and the expression patterns in different developmental stages and tissues, as well as in response to both thermal and 20-hydroxyecdysone (20E) exposures, were examined using real time quantitative PCR. The open reading frames (ORFs) of six sHsps are 453, 489, 537, 543, 567 and 630bp in length, encoding proteins with molecular weights of 17.7, 18.4, 20.4, 20.6, 21.6 and 23.8kDa, respectively. BdHsp18.4 and BdHsp20.4 maintained lower expression levels in both eggs and larvae, whereas remarkably up-regulated after the larval-pupal transformation, suggesting that these two sHsps may be involved in metamorphosis. Significant tissue specificity exists among sHsps: the highest expression of BdHsp20.6 and BdHsp23.8 in the Malpighian tubules and ovary, respectively, versus a peak in the fat body for others. BdHsp20.4 and BdHsp20.6 were significantly up-regulated by thermal stress. In contrast, BdHsp18.4 and BdHsp23.8 reacted only to heat stress. BdHsp17.7 and BdHsp21.6 were insensitive to both heat and cold stresses. The degree of sHsps response depends on intensity of 20E treatment, i.e., dose and time. These results strongly suggest functional differentiation within the sHsp subfamily in B. dorsalis. The physiological function of sHsp members under thermal stress and normal growth remains the subjects of further investigation.

Characteristics and expression of genes encoding two small heat shock protein genes lacking introns from Chilo suppressalis.

Small heat shock proteins (sHSPs) constitute a large, diverse, and functionally uncharacterized family of heat shock proteins. To gain insight regarding the function of sHSPs in insects, we identified genes encoding two sHSPs, Cshsp22.9b and Cshsp24.3, from the rice pest Chilo suppressalis. The cDNAs of Cshsp22.9b and Cshsp24.3 encoded proteins of 206 and 216 amino acids with isoelectric points of 5.79 and 9.28, respectively. Further characterization indicated that both Cshsp22.9b and Cshsp24.3 lacked introns. Real-time quantitative PCR indicated that Cshsp22.9b and Cshsp24.3 were expressed at higher levels within the fat body as compared to other tissues (head, epidermis, foregut, midgut, hindgut, Malpighian tubules, and hemocytes). Expression of Cshsp22.9b and Cshsp24.3 was lowest in the hindgut and Malpighian tubules, respectively. Cshsp22.9b and Cshsp24.3 showed identical patterns in response to thermal stress from -11 to 43°C, and both genes were up-regulated by hot and cold temperatures. The mRNA (messenger ribonucleic acid) expression levels of Cshsp22.9b (KY701308) and Cshsp24.3 (KY701309) were highest after a 2-h exposure at 39°C and started to decline at 42°C. In response to cold temperatures, both Cshsp22.9b and Cshsp24.3 showed maximal expression after a 2-h exposure to -3°C. The two Cshsps were more responsive to hot than cold temperature stress and were not induced by mildly cold or warm temperatures. In conclusion, Cshsp22.9b and Cshsp24.3 could play a very important role in the regulation of physiological activities in C. suppressalis that are impacted by environmental stimuli.

Heterologous Expression of theCarrot Hsp17.7 gene Increased Growth, Cell Viability, and Protein Solubility in Transformed Yeast (Saccharomyces cerevisiae) under Heat, Cold, Acid, and Osmotic Stress Conditions.

In industrial fermentation of yeast (Saccharomyces cerevisiae), culture conditions are often modified from the optimal growth conditions of the cells to maintain large-scale cultures and/or to increase recombinant protein production. However, altered growth conditions can be stressful to yeast cells resulting in reduced cell growth and viability. In this study, a small heat shock protein gene from carrot (Daucus carota L.), Hsp17.7, was inserted into the yeast genome via homologous recombination to increase tolerance to stress conditions that can occur during industrial culture. A DNA construct, Translational elongation factor gene promoter-carrot Hsp17.7 gene-Phosphoribosyl-anthranilate isomerase gene (an auxotrophic marker), was generated by a series of PCRs and introduced into the chromosome IV of the yeast genome. Immunoblot analysis showed that carrot Hsp17.7 accumulated in the transformed yeast cell lines. Growth rates and cell viability of these cell lines were higher than control cell lines under heat, cold, acid, and hyperosmotic stress conditions. Soluble protein levels were higher in the transgenic cell lines than control cell lines under heat and cold conditions, suggesting the molecular chaperone function of the recombinant Hsp17.7. This study showed that a recombinant DNA construct containing a HSP gene from carrot was successfully expressed in yeast by homologous recombination and increased tolerances to abiotic stress conditions.

Heterologous expression of three Camellia sinensis small heat shock protein genes confers temperature stress tolerance in yeast and Arabidopsis thaliana.

CsHSP17.7, CsHSP18.1, and CsHSP21.8 expressions are induced by heat and cold stresses, and CsHSP overexpression confers tolerance to heat and cold stresses in transgenic Pichia pastoris and Arabidopsis thaliana. Small heat shock proteins (sHSPs) are crucial for protecting plants against biotic and abiotic stresses, especially heat stress. However, knowledge concerning the functions of Camellia sinensis sHSP in heat and cold stresses remains poorly understood. In this study, three C. sinensis sHSP genes (i.e., CsHSP17.7, CsHSP18.1, and CsHSP21.8) were isolated and characterized using suppression subtractive hybridization (SSH) technology. The CsHSPs expression levels in C. sinensis leaves were significantly up-regulated by heat and cold stresses. Phylogenetic analyses revealed that CsHSP17.7, CsHSP18.1, and CsHSP21.8 belong to sHSP Classes I, II, and IV, respectively. Heterologous expression of the three CsHSP genes in Pichia pastoris cells enhanced heat and cold stress tolerance. When exposed to heat and cold treatments, transgenic Arabidopsis thaliana plants overexpressing CsHSP17.7, CsHSP18.1, and CsHSP21.8 had lower malondialdehyde contents, ion leakage, higher proline contents, and transcript levels of stress-related genes (e.g., AtPOD, AtAPX1, AtP5CS2, and AtProT1) compared with the control line. In addition, improved seed germination vigor was also observed in the CsHSP-overexpressing seeds under heat stress. Taken together, our results suggest that the three identified CsHSP genes play key roles in heat and cold tolerance.

Molecular characterisation and expression analysis of a sHSP gene involved in heat shock treatment-induced chilling tolerance in highbush blueberry fruit

ABSTRACTPlant small heat shock proteins (sHSPs) are abundant and diverse proteins involved in various stresses. In this study, a small heat shock protein gene (VcHSP17.7) encoding a 160-amino acid protein was isolated from the highbush blueberry (Vaccinium corymbosum L.). Sequence alignment and phylogenetic analysis indicated that the VcHSP17.7 is a member of the plant cytosolic class II sHSPs; this finding was supported by the subcellular localisation pattern of a VcHSP17.7-YFP fusion protein. Quantitative real-time PCR (qRT-PCR) showed that the VcHSP17.7 gene was expressed in all analysed tissues. Heat shock experiments were conducted using mature blueberry fruit. The anthocyanin content was lower in heat shock-treated fruit than in control fruit after 14 days of storage at 2°C. The malondialdehyde (MDA) content was lower in heat shock-treated fruit than in control fruit during subsequent low-temperature storage, which indicates that heat shock treatment may have alleviated chilling injury (CI) and enhanced the chilling tolerance of the blueberry fruit. Meanwhile, the expression of VcHSP17.7 in blueberry fruit was induced by heat shock treatment and gradually increased during subsequent low-temperature storage. These results suggest that VcHSP17.7 might be involved in the chilling tolerance of blueberry fruit caused by heat shock treatment.

Physio-Biochemical and Molecular Responses in Transgenic Cotton under Drought Stress

Drought decreases the growth and productivity in cotton. Heat shock proteins accumulate in plants under water stress to protect the biochemical and physiological processes at the molecular level. In this study, plants of T2 segregating generation of transgenic cotton, containing small heat shock protein gene (GHSP26) was compared with wild type plants for biochemical, physiological and molecular responses under different periods of drought stress. Transgenic plants accumulated 30% higher proline content than the wild type. Lipid peroxidation activity was reduced in transgenic plants which showed that the drought tolerance efficiency has been improved. Leaf relative water content was 69% and 45% in transgenic and wild-type plants, respectively at 10-day drought stress. Similarly, transgenic plants showed better performance for photosynthesis, stomatal conductance, transpiration and osmotic potential as compared to wild type. Real-time quantitative PCR of GHSP26 and some other drought responsive genes such as Gh-POD, Gh-RuBisCO, Gh-LHCP PSII, Gh-PIP, Gh-TPS and Gh-LEA have supported the higher expression and proved drought tolerance in transgenic plants. The overexpression of GHSP26 in transgenic plants improved the biochemical such as proline content and lipid peroxidation activity and physiological parameters like photosynthesis, osmotic potential and water related attributes. Hence, this study may be extended for selection of homozygous lines and breeding to improve the drought tolerance activity in plants.

Early Heat Shock Protein Response and Selection of Reference Genes in Arabidopsis thaliana Seedlings Subjected to Marine Fuel Contamination

Strategies for management of damaged environments can benefit from understanding of how early petrochemical pollution affect living organisms. One of the general responses to environmental stress in plants is mediated by the regulatory network of heat shock proteins (HSP). Arabidopsis thaliana is a model plant for genetic studies, and laboratory experiments with this species might be informative for predicting analogous responses to toxicants in other species. Here, Arabidopsis seedlings were exposed to time-varying contamination (up to 24 h) with the water soluble fraction of MF380 marine fuel (WSF-MF380). An accurate estimation of expression differences in HSP genes was obtained by real-time quantitative polymerase chain reaction (qPCR). After a thorough selection and validation of reference genes, two gene pairs were found to be stably expressed across control and WSF-MF380-treated samples and were used as normalization factors. Next, we evaluated the normalized expression of five HSP genes in response to the time-varying WSF-MF380 contamination. Four HSPs presented a significant increase in gene expression, which suggests that they might be tested as biomarkers for early exposure to petrochemical compounds. While a nearly immediate response (3 h after contamination) was found for HSP90.1 and two small HSP genes localized in the mitochondria (sHSP23.5 and sHSP26.5), a slightly later response (20 h) was observed for a third small HSP with a cytoplasmic/nuclear localization (sHSP18.2). Overall, these expression changes suggest the existence of a genetic cross-talk between canonical regulatory networks of HSPs and the cellular response to non-heat stress factors, such as marine oil contamination.

Isolation and allelic characterization of finger millet (Eleusine coracana L.) small heat shock protein EcHSP17.8 for stress tolerance

Finger millet (Eleusine coracana L.) commonly known as ragi is an orphan cereal crop having significant climatesmart stress adaptation system. Apart from various abiotic stresses worldwide, heat stress is the major limiting constraint for crop yield and the loss of crop yield due to heat stress is increasing every year. Hence, to generate better heat tolerant crop plants there is a need to mine superior alleles. In this regard, efforts were made to isolate and characterize role of small heat shock protein gene HSP17.8 from finger millet. Here, we obtained an EST of having Hsp20 domain based on the expression analysis under heat stress and identified complete gene using homology-based analysis. The obtained gene was named as EcHSP17.8 based on its molecular weight. The complete Open Reading Frame (ORF) of EcHSP17.8 is of 489 bp with peptide length of 162 aa. Expression analysis of this gene in finger millet revealed its maximum expression in roots tissue. This gene was found to be induced by heat (42°C), desiccation, NaCl (250 mM), mannitol (200 mM) and oxidative stress; however maximum upregulation was observed under heat stress up to 40 folds. Expression analysis of this gene in 16 finger millet genotypes under heat stress revealed its maximum expression after 2h which remained upregulated up to 4h showing it as early responsive gene under heat stress. As this gene is well characterized for stress tolerance in other crops, the alleles of HSP17.8 from crop like finger millet can be used as better candidate for generating heat tolerant crop plants.

Maternal loading of a small heat shock protein increases embryo thermal tolerance in Drosophila melanogaster.

Maternal investment is likely to have direct effects on offspring survival. In oviparous animals whose embryos are exposed to the external environment, maternal provisioning of molecular factors like mRNAs and proteins may help embryos cope with sudden changes in the environment. Here, we sought to modify the maternal mRNA contribution to offspring embryos and test for maternal effects on acute thermal tolerance in early embryos of Drosophila melanogaster We drove in vivo overexpression of a small heat shock protein gene (Hsp23) in female ovaries and measured the effects of acute thermal stress on offspring embryonic survival and larval development. We report that overexpression of the Hsp23 gene in female ovaries produced offspring embryos with increased thermal tolerance. We also found that brief heat stress in the early embryonic stage (0-1 h old) caused decreased larval performance later in life (5-10 days old), as indexed by pupation height. Maternal overexpression of Hsp23 protected embryos against this heat-induced defect in larval performance. Our data demonstrate that transient products of single genes have large and lasting effects on whole-organism environmental tolerance. Further, our results suggest that maternal effects have a profound impact on offspring survival in the context of thermal variability.

Cadmium alters the expression of small heat shock protein genes in the aquatic midge Chironomus riparius

Cadmium (Cd) is a widespread and highly toxic heavy metal of particular ecotoxicological relevance for aquatic ecosystems. It occurs naturally in the environment but is also an industrial pollutant with extensively researched carcinogenic potentials. Heat shock proteins (HSPs) are chaperones that play an important role in maintaining protein homeostasis under stress conditions. Small heat shock proteins (sHSPs) comprise the most diverse group of the HSPs family. They are expressed both constitutively and by stress-induction. The midge Chironomus riparius is widely used as a test species in aquatic toxicology. In the present study, Reverse Transcription Polymerase Chain Reaction (RT-PCR) was used to evaluate the effects of acute Cd exposure to the expression profile of seven shsp genes (hsp17, hsp21, hsp22, hsp23, hsp24, hsp27, and hsp34) in C. riparius larvae. Results show a specific pattern of response with a rapid response by hsp27, which was downregulated at 2–6 h, while the rest of the shsp genes remained unaltered except for hsp17 at 2 h, which was upregulated. However, at 24 h of exposure are observed high levels of hsp23, hsp24, hsp27, and hsp34 transcription while hsp22 mRNA levels were downregulated and hsp17 and hsp21 remained unaltered. These changes in gene expression suggest a functional diversity between the sHSPs in the cellular response to heavy metal stress. The differential pattern in comparison with heat shock supports a specific profile depending on the stress supporting the use of shsp genes as suitable biomarkers for ecotoxicological studies on aquatic systems.

Effect of long term heat stress and dietary restriction on the expression of small heat shock protein (sHSP) genes in rat liver tissue

Effect of long term heat stress and dietary restriction on the expression of small heat shock protein (sHSP) genes in rat liver tissue

DFOXO Activates Large and Small Heat Shock Protein Genes in Response to Oxidative Stress to Maintain Proteostasis in Drosophila

Maintaining protein homeostasis is critical for survival at the cellular and organismal level (Morimoto, R. I. (2011) Cold Spring Harb. Symp. Quant. Biol. 76, 91-99). Cells express a family of molecular chaperones, the heat shock proteins, during times of oxidative stress to protect against proteotoxicity. We have identified a second stress responsive transcription factor, dFOXO, that works alongside the heat shock transcription factor to activate transcription of both the small heat shock protein and the large heat shock protein genes. This expression likely protects cells from protein misfolding associated with oxidative stress. Here we identify the regions of the Hsp70 promoter essential for FOXO-dependent transcription using in vitro methods and find a physiological role for FOXO-dependent expression of heat shock proteins in vivo.

Expression analysis of two reverse duplicated small heat shock protein genes in rice (Oryza sativa L.)

The small heat shock protein (sHSP) chaperones are required for protecting cellular proteins from damage, as well as re-folding denatured proteins. This study was carried out to investigate the temporal-spatial expression patterns of two sHSP genes in rice. These two genes, named as Os16.9A and Os16.9B, are reverse duplicated genes that adjacently located on chromosome 1 and probably share the same or overlapping DNA region as a promoter. The interval sequence between the start codons of the two genes which are transcribed in opposite directions is only about 2.6 kb. Semi-quantitative RT-PCR was carried out to detect the expression of the two genes under normal growth conditions and different stress conditions. The expression patterns of the two genes were in detail investigated by using β-glucuronidase (GUS) reporter gene fusion system. Results showed that heat shock stress can induce high level expression of the two genes. Under normal growth conditions, Os16.9A and Os16.9B expressed in vegetative organs and young panicles. GUS staining combined with cytological observations showed that the two genes expressed mainly in the vascular tissues of roots, stems and young panicles, implicating that Os16.9A and Os16.9B play important roles not only for heat shock response, but also for normal development in rice.

Comparative analysis of nine different small heat shock protein gene promoters in Oryza sativa L. subsp. indica

Small heat shock proteins (sHSPs), the intra cellular chaperons respond to high temperatures and other stresses, are expressed under the control of the heat shock factors, which recognize highly conserved cis-regulatory elements (CREs). The present study was designed to identify and compare nine different sHSP gene promoters from rice on the basis of their location on chromosomes, sequence similarities, evolutionary relationship and putative binding sites of transcription factors. Additionally, they were analyzed for their functional importance in the regulation of sHSP genes. For this, 5′ upstream region of nine sHSP genes from Oryza sativa was amplified, sequenced and analyzed. RFLP mapping approach using molecular linkage maps of three-way cross-population showed that OsHSP 16.9A, OsHSP 16.9B, OsHSP 16.9C and OsHSP C-II genes were clustered on the short arm of chromosome 1 with a 1238-bp overlapped sequence between OsHSP 16.9A and OsHSP 16.9B, whereas OsHSP 17.3, OsHSP 17.7, OsHSP 17.9, OsHSP 18.0 and OsHSP 26 genes were clustered on chromosome 3 with a 420-bp overlapped sequence between OsHSP 17.3 and OsHSP 18.0. Evolutionary analysis revealed that sHSPs that clustered on different chromosomes in the same specie showed high sequence similarity and evolved prior to the divergence of their localization. Eighteen CREs remained conserved among all sHSP gene promoters during evolution. These common regulatory elements are related to disease resistance, hormonal responses, light responses, seed storage proteins, tissue-specific expression, pollen development and other environmental stresses. Moreover, CCAATBOX1 was also found in all sHSP gene promoters involved in the regulation of heat shock response. Conclusively, promoter region of sHSP genes seems to be capable of driving genes expression during stress and genetic manipulation might be helpful in producing better quality rice.

Molecular cloning and characterization of the MsHSP17.7 gene from Medicago sativa L.

Heat shock proteins (HSPs) are ubiquitous protective proteins that play crucial roles in plant development and adaptation to stress, and the aim of this study is to characterize the HSP gene in alfalfa. Here we isolated a small heat shock protein gene (MsHSP17.7) from alfalfa by homology-based cloning. MsHSP17.7 contains a 477-bp open reading frame and encodes a protein of 17.70-kDa. The amino acid sequence shares high identity with MtHSP (93.98 %), PsHSP17.1 (83.13 %), GmHSP17.9 (74.10 %) and SlHSP17.6 (79.25 %). Phylogenetic analysis revealed that MsHSP17.7 belongs to the group of cytosolic class II small heat shock proteins (sHSP), and likely localizes to the cytoplasm. Quantitative RT-PCR indicated that MsHSP17.7 was induced by heat shock, high salinity, peroxide and drought stress. Prokaryotic expression indicated that the salt and peroxide tolerance of Escherichia coli was remarkably enhanced. Transgenic Arabidopsis plants overexpressing MsHSP17.7 exhibited increased root length of transgenic Arabidopsis lines under salt stress compared to the wild-type line. The malondialdehyde (MDA) levels in the transgenic lines were significantly lower than in wild-type, although proline levels were similar between transgenic and wild-type lines. MsHSP17.7 was induced by heat shock, high salinity, oxidative stress and drought stress. Overexpression analysis suggests that MsHSP17.7 might play a key role in response to high salinity stress.Electronic supplementary materialThe online version of this article (doi:10.1007/s11033-016-4008-9) contains supplementary material, which is available to authorized users.

Transcriptome analysis reveals translational regulation in barley microspore-derived embryogenic callus under salt stress.

Transcriptome analysis of barley embryogenic callus from isolated microspore culture under salt stress uncovered a role of translation inhibition and selective activation of stress-specific proteins in cellular defense. Soil salinity is one of the major abiotic stresses which constrains the plant growth and reduces the productivity of field crops. In this study, it was observed that the salt stress in barley isolated microspore culture impacted not only on the quantity of embryogenic callus but also on the quality for later differentiation. The barley microspore-derived embryogenic callus, a transient intermediate form linked cells and plants, was employed for a global transcriptome analysis by RNA sequencing to provide new insights into the cellular adaptation or acclimation to stress. A total of 596 differentially expressed genes (DEGs) were identified, in which 123 DEGs were up-regulated and 473 DEGs were down-regulated in the embryogenic callus produced from microspore culture under salt stress as compared to the control conditions. KEGG pathway analysis identified 'translation' (27 DEGs; 12.56%) as the largest group and followed by 'folding, sorting and degradation' (25 DEGs; 11.63%) in 215 mapped metabolic pathways. The results of RNA-Seq data and quantitative real-time polymerase chain reaction validation showed that the genes related to translation regulation (such as eIF1A, RPLP0, RPLP2, VARS) were down-regulated to control general protein synthesis, and the genes related to endoplasmic reticulum stress response (such as small heat shock protein genes) were selectively up-regulated against protein denaturing during microspore embryogenesis under continuous salt stress. These transcriptional remodeling might affect the essential protein synthesis for the cell development to fulfill totipotency under salt stress.