Physiological and transcriptional responses to heat stress in a typical phenotype of Pinellia ternata

Pinellia ternata is an important medicinal plant,and its growth and development are easily threatened by high temperature.In this study,comprehensive research on physiological,cytological and transcriptional responses to different levels of heat stress were conducted on a typical phenotype of P.ternata.First,P.ternata exhibited tolerance to the increased temperature,which was supported by normal growing leaves,as well as decreased and sustained photosynthetic parameters.Severe stress aggravated the damages,and P.ternata displayed an obvious leaf senescence phenotype,with significantly increased SOD and POD activities(46%and 213%).In addition,mesophyll cells were seriously damaged,chloroplast thylakoid was fuzzy,grana lamellae and stroma lamellae were obviously broken,and grana thylakoids were stacked,resulting in a dramatically declined photosynthetic rate(74.6%).Moreover,a total of 16808 genes were significantly differential expressed during this process,most of which were involved in photosynthesis,transmembrane transporter activity and plastid metabolism.The number of differentially expressed transcription factors in MYB and bHLH families was the largest,indicating that these genes might participate in heat stress response in P.ternata.These findings provide insight into the response to high temperature and facilitate the standardized cultivation of P.ternata.

Arabidopsis transcriptional response to extracellular Ca^(2+) depletion involves a transient rise in cytosolic Ca^(2+)

Ecological evidence indicates a worldwide trend of dramatically decreased soil Ca2＋ levels caused by increased acid deposition and massive timber harvesting. Little is known about the genetic and cellular mechanism of plants＇ responses to Ca2＋ depletion. In this study, transcriptional profiling analysis helped identify multiple extracellular Ca2＋ （[Ca2＋]ext） depletion-responsive genes in Arabidopsis thaliana L., many of which are involved in response to other environmental stresses. Interestingly, a group of genes encoding putative cytosolic Ca2＋ （[Ca2＋]cyt） sensors were significantly upregulated, implying that [Ca2＋]cyt has a role in sensing [Ca2＋]ext depletion. Consistent with this observation, [Ca2＋]ext depletion stimulated a transient rise in [Ca2＋]cyt that was negatively influenced by [K＋]ext, suggesting the involvement of a membrane potential-sensitive component. The [Ca2＋]cyt response to [Ca2＋]ext depletion was significantly desensitized after the initial treatment, which is typical of a receptor-mediated signaling event. The response was insensitive to an animal Ca2＋ sensor antagonist, but was suppressed by neomycin, an inhibitor of phospholipase C. Gd3＋, an inhibitor of Ca2＋ channels, suppressed the [Ca2＋]ext-triggered rise in [Ca2＋]cyt and downstream changes in gene expression. Taken together, this study demonstrates that [Ca2＋]cyt plays an important role in the putative receptor-mediated cellular and transcriptional response to [Ca2＋]ext depletion of plant cells.

Transcriptional activation of glucose transporter 1 in orthodontic tooth movement-associated mechanical response

The interplay between mechanoresponses and a broad range of fundamental biological processes, such as cell cycle progression,growth and differentiation, has been extensively investigated. However, metabolic regulation in mechanobiology remains largely unexplored. Here, we identified glucose transporter 1(GLUT1)—the primary glucose transporter in various cells—as a novel mechanosensitive gene in orthodontic tooth movement(OTM). Using an in vivo rat OTM model, we demonstrated the specific induction of Glut1 proteins on the compressive side of a physically strained periodontal ligament. This transcriptional activation could be recapitulated in in vitro cultured human periodontal ligament cells(PDLCs), showing a time-and dose-dependent mechanoresponse. Importantly, application of GLUT1 specific inhibitor WZB117 greatly suppressed the efficiency of orthodontic tooth movement in a mouse OTM model, and this reduction was associated with a decline in osteoclastic activities. A mechanistic study suggested that GLUT1 inhibition affected the receptor activator for nuclear factor-κ B Ligand(RANKL)/osteoprotegerin(OPG)system by impairing compressive force-mediated RANKL upregulation. Consistently, pretreatment of PDLCs with WZB117 severely impeded the osteoclastic differentiation of co-cultured RAW264.7 cells. Further biochemical analysis indicated mutual regulation between GLUT1 and the MEK/ERK cascade to relay potential communication between glucose uptake and mechanical stress response. Together, these cross-species experiments revealed the transcriptional activation of GLUT1 as a novel and conserved linkage between metabolism and bone remodelling.

Poly(ADP-ribosyl)ation of Apoptosis Antagonizing Transcription Factor Involved in Hydroquinone-Induced DNA Damage Response

The molecular mechanism of DNA damage induced by hydroquinone （HQ） remains unclear. Poly（ADP-ribose） polymerase-1 （PARP-1） usually works as a DNA damage sensor, and hence, it is possible that PARP-1 is involved in the DNA damage response induced by HQ. In TK6 cells treated with HQ, PARP activity as well as the expression of apoptosis antagonizing transcription factor （AATF）, PARP-1, and phosphorylated H2AX （v-H2AX） were maximum at 0.5 h, 6 h, 3 h, and 3 h, respectively. To explore the detailed mechanisms underlying the prompt DNA repair reaction, the above indicators were investigated in PARP-l-silenced cells. PARP activity and expression of AATF and PARP-1 decreased to 36%, 32%, and 33%, respectively, in the cells; however, y-H2AX expression increased to 265%. Co-immunoprecipitation （co-IP） assays were employed to determine whether PARP-1 and AATF formed protein complexes. The interaction between these proteins together with the results from IP assays and confocal microscopy indicated that poly（ADP-ribosyl）ation {PARylation） regulated AATF expression, in conclusion, PARP-1 was involved in the DNA damage repair induced by HQ via increasing the accumulation of AATF through PARylation.

Edaravone protects against oxygen-glucose-serum deprivation/restoration-induced apoptosis in spinal cord astrocytes by inhibiting integrated stress response

We previously found that oxygen-glucose-serum deprivation/restoration（OGSD/R） induces apoptosis of spinal cord astrocytes, possibly via caspase-12 and the integrated stress response, which involves protein kinase R-like endoplasmic reticulum kinase（PERK）, eukaryotic initiation factor 2-alpha（eIF2α） and activating transcription factor 4（ATF4）. We hypothesized that edaravone, a low molecular weight, lipophilic free radical scavenger, would reduce OGSD/R-induced apoptosis of spinal cord astrocytes. To test this, we established primary cultures of rat astrocytes, and exposed them to 8 hours/6 hours of OGSD/R with or without edaravone（0.1, 1, 10, 100 μM） treatment. We found that 100 μM of edaravone significantly suppressed astrocyte apoptosis and inhibited the release of reactive oxygen species. It also inhibited the activation of caspase-12 and caspase-3, and reduced the expression of homologous CCAAT/enhancer binding protein, phosphorylated（p）-PERK, p-eIF2α, and ATF4. These results point to a new use of an established drug in the prevention of OGSD/R-mediated spinal cord astrocyte apoptosis via the integrated stress response.

Comparative transcriptome analysis of atmospheric pressure cold plasma enhanced early seedling growth in Arabidopsis thaliana

The stimulatory effects of atmospheric pressure cold plasma(APCP)on plant growth have attracted much attention due to its great potential as a new approach to increase crop growth and production.However,the transcriptome changes of plants induced by APCP treatment are unknown.Herein,the comparative transcriptome analysis was performed to identify the transcriptional response of Arabidopsis thaliana seedlings to APCP.Results showed that APCP exhibited a dual effect(stimulation or inhibition)on Arabidopsis seedling growth dependent on the treatment time and the maximum stimulatory effects were achieved by 1 min APCP treatment.The metabolic analysis of amino acid,glutathione(GSH)and phytohormone demonstrated that 1 min APCP treatment decreased most amino acids concentrations in Arabidopsis seedling,while the accumulations of GSH,gibberellins and cytokinin were significantly increased.The RNA-Seq analysis showed that a total of218 differentially expressed genes(DEGs)were identified in 1 min APCP-treated seedlings versus the control,including 20 up-regulated and 198 down-regulated genes.The DEGs were enriched in pathways related to GSH metabolism,mitogen-activated protein kinase(MAPK)signaling transduction and plant resistance against pathogens.Moreover,most of the DEGs were defense,stimuli or stressresponsive genes and encoded proteins with oxidoreductase activity.Expression determination of six randomly selected DEGs by quantitative real-time PCR demonstrated similar pattern with the RNASeq data.These results indicated that the moderate APCP treatment may regulate the expression of stimuli/stress-responsive genes involved in GSH,phytohormone/amino metabolism and plant defense against pathogens via MAPK signal transduction pathway,accordingly enhance Arabidopsis seedling growth.This study provides a theoretical basis for the application of APCP in agriculture.

Improvement of Copper-inducible Gene Expression System for Plant

The copper-regulated gene expression system has been developed to control spacial and temporal expression of transgene in plant. It comprises two parts: (1) ace I gene encoding copper-responsive transcription factor under the control of a constitutive or organ-specific promoter, and (2) a gene of interest under the control of a chimeric promoter consisting of the CaMV 35S (-90 to +8) promoter linked to the metal responsive element (MRE) carrying activating copper-metallothionein expression (ACE1)-binding sites. Here, the effectiveness of two different ACE1-binding cis -elements which derive from 5'-regulatory region of yeast metallothionein gene was investigated in transgenic tobacco (Nicotiana tabacum L. cv. W38). The results revealed that the MRE (-210 to -126) could increase the system inducibility by 50% - 100% compared with the previously reported MRE (-148 to -105). It is potential to use the copper-inducible system to control valuable gene traits in plant biotechnology.

STOP1 regulates CCX1-mediated Ca^(2+)homeostasis for plant adaptation to Ca^(2+)deprivation

Calcium(Ca)is essential for plant growth and stress adaptation,yet its availability is often limited in acidic soils,posing a major threat to crop production.Understanding the intricate mechanisms orchestrating plant adaptation to Ca deficiency remains elusive.Here,we show that the Ca deficiency-enhanced nuclear accumulation of the transcription factor SENSITIVE TO PROTON RHIZOTOXICITY 1(STOP1)in Arabidopsis thaliana confers tolerance to Ca deprivation,with the global transcriptional responses triggered by Ca deprivation largely impaired in the stop1 mutant.Notably,STOP1 activates the Ca deprivation-induced expression of CATION/Ca^(2+)EXCHANGER 1(CCX1)by directly binding to its promoter region,which facilitates Ca^(2+)efflux from endoplasmic reticulum to cytosol to maintain Ca homeostasis.Consequently,the constitutive expression of CCX1 in the stop1 mutant partially rescues the Ca deficiency phenotype by increasing Ca content in the shoots.These findings uncover the pivotal role of the STOP1-CCX1 axis in plant adaptation to low Ca,offering alternative manipulating strategies to improve plant Ca nutrition in acidic soils and extending our understanding of the multifaceted role of STOP1.

WRKY transcription factors in plant responses to stresses

The WRKY gene family is among the largest families of transcription factors （TFs） in higher plants. By regulating the plant hormone signal transduction pathway, these TFs play critical roles in some plant processes in response to biotic and abiotic stress, Various bodies of research have demonstrated the important biological functions of WRKY TFs in plant response to different kinds of biotic and abiotic stresses and working mecha- nisms. However, very li2ttle summarization has been done to review their research progress. Not iust important TFs function in plant response to biotic and abiotic stresses, WRKY also participates in carbohydrate synthesis, senes- cence, development, and secondary metabolites synthesis. WRKY proteins can bind to W-box （TGACC （A/T）） in the promoter of its target genes and activate or repress the expression of downstream genes to regulate their stress response. Moreover, WRKY proteins can interact with other TFs to regulate plant defensive responses. In the present review, we focus on the structural characteristics of WRKY TFs and the research progress on their functions in plant responses to a variety of stresses.

Transcriptional activation of Nlp by estrogen-ERa in breast cancer

Estrogen Receptor-a （ERa） is the key transcription factor that regulates cell proliferation and homeosta- sis. In this pathway, estrogen plays an important role in genomic instability and cell cycle regulation pro- cesses and the mechanisms of its action are multifaceted. In this study, we showed that estrogen regulates genomic instability through promoting the expression of Nip, a BRCAl-associated centrosomal protein which is involved in microtubule nucleation, spindle formation, chromosomal missegregation and abnormal cytokinesis. We demonstrated that the expression of Nip is strongly associated with ERa and FOXAI level in clinical breast cancer samples with poor clinical outcomes to breast cancer patients. Addition of estrogen in the ER-positive breast cancer cells resulted in elevation of NLP mRNA. Significantly, we identified that estrogen-ERa is capable of regulating Nlp expression through specifically binding ERa to the proximal region and the Estrogen Responsive Elements （ERE） enhancer in the distal region of NLP gene. Reporter assays demonstrated that estrogen directly activated Nlp promoter. ChIP assay results showed that E2-ERa directly bound to the EREs of Nip. Therefore, overexpression of Nip in breast cancer exhibits a hormone-dependent pattern, and estrogen participates in the regulation of genome instability and cell cycle in breast cancer cells partially through transcriptional activation of NLP gene. Overexpression of Nlp enhances the malignant progression of ERa-positive breast cancer cells in vitro, whereas knockdown of Nip suppresses this biological effects in ERa-positive breast cancer ceils. ERa/NIp axis may serve as a promising target against breast cancer.

OsDREB4 Genes in Rice Encode AP2-Containing Proteins that Bind Specifically to the Dehydration-Responsive Element

Most dehydration-responsive element-binding (DREB) factors interact specifically with the dehydration-responsive element (DRE) and control the expression of many stress-inducible genes in Arabidopsis. In rice (Oryza sativa L. cv. Lansheng), we cloned three DREB homologs: OsDREB1-1, OsDREB4- 1, and OsDREB4-2. The deduced amino acid sequences revealed that each protein contained a potential nuclear localization signal, an AP2 DNA-binding domain, and a possible acidic activation domain. The yeast one-hybrid assay indicated that both OsDREB4-1 and OsDREB4-2 proteins specifically bound to DRE and activated expression of the dual reporter genes of histidine (HIS3) and galactosidase (LacZ). In rice seedlings,expression of OsDREB4-1 was induced by dehydration and high salt, whereas OsDREB1-1 and OsDREB4-2 were expressed constitutively. Under normal growth conditions, OsDREB1-1 was expressed strongly in the leaf, sheath, and spike, was expressed relatively weak in the stem and only faintly expressed in the roots,whereas expression of transcripts of OsDREB4-1 and OsDREB4-2 was higher in the roots, stem, and spike,lower in the leaf, and undetectable in the sheath. Together, these results imply that expression of the OsDREB genes could be controlled by specific aspects of differentiation or development. Thus, OsDREB4-1 could function as a trans-acting factor in the DRE/DREB regulated stress-responsive pathway.

Integrated biomarkers in wild crucian carp for early warning of water quality in Hun River, North China

Metabolizing enzymes play important roles in the detoxification of various pollutants in aquatic organisms, thereby they can also be used to provide early-warning signals of environmental risks. Real-time quantitative reverse-transcription polymerase chain reaction assays were developed to quantify cytochrome P450 1A （CYP1A）, superoxide dismutase （SOD）, glutathione peroxidase （GPx）, catalase （CAT）, and glutathione-S-transferase （GST） in crucian carp （Carassius auratus）. The methods were then used to detect the respective mRNA expression levels in liver tissue in wild crucian carp from the Hun River, North China. CYP1A mRNA expression was significantly up-regulated in fish from stations $5, $6, and $8 （p 〈 0.05）. SOD mRNA expression was significantly down-regulated in downstream areas relative to fish from upstream sites （p 〈 0.05）; GPx and CAT mRNA expression levels were also down-regulated at $9 （p 〈 0.05）. In contrast, GST mRNA expression showed no obvious change between fish collected from up- or downstream areas of the river. Finally, an integrated biomarker response was used to evaluate the integrated impact of pollutants in the Hun River and allow better comprehension of the real toxicological risk of these investigated sites.