Comparative transcriptome provides new insights to the molecular regulation of olive trees to chilling stress.

Olive (Olea europaea. L), an economically important oil-producing crop, is sensitive to low temperature, which severely limits its productivity and geographical distribution. However, the underlying mechanism of cold tolerance in olive remains elusive. In this study, a chilling experiment (4 °C) on the living saplings of two olive cultivars revealed that O. europaea cv. Arbequina showed stronger cold tolerance with greater photosynthetic activity compared to O. europaea cv. Leccino. Transcriptome analyses revealed that early light-inducible protein 1 (ELIP1), the main regulator for chlorophyll synthesis, is dramatically induced to protect the photosynthesis at low temperatures. Furthermore, weighted gene co-expression network analysis (WGCNA), yeast one-hybrid (Y1H), and luciferase (LUC) assays demonstrated that transcription factor bHLH66 serves as an important regulator of ELIP1 transcription by binding to the G-box motif in the promoter. Taken together, our research revealed a novel transcriptional module consisting of bHLH66- ELIP1 in the adaptation of olive trees to cold stress.

An alternative 3' splice site of PeuHKT1;3 improves the response to salt stress through enhancing affinity to K+ in Populus.

Alternative splicing (AS) serves as a crucial post-transcriptional regulator in plants that contributes to the resistance to salt stress. However, the underlying mechanism is largely unknown. In this research, we identified an important AS transcript in Populus euphratica, PeuHKT1:3a, generated by alternative 3' splice site splicing mode that resulted in the removal of 252 bases at the 5' end of the first exon in PeuHKT1:3. Protein sequence comparison showed that the site of AS occurred in PeuHKT1:3 is located at a crucial Ser residue within the first pore-loop domain, which leads to inefficient K+ transport in HKT I-type transporters. Expressing PeuHKT1;3a in an axt3 mutant yeast strain can effectively compensate for the lack of intracellular K+, whereas the expression of PeuHKT1;3 cannot yield the effect. Furthermore, in transgenic Arabidopsis and poplar plants, it was observed that lines expressing PeuHKT1;3a exhibited greater salt tolerance compared to those expressing the PeuHKT1;3 strain. Analysis of ion content and flux demonstrated that the transgenic PeuHKT1;3a line exhibited significantly higher K+ content compared to the PeuHKT1;3 line, while there was no significant difference in Na+ content. In conclusion, our findings revealed that AS can give rise to novel variants of HKT I-type proteins in P. euphratica with modified K+ selectivity to keep a higher K+/Na+ ratio to enhanced salt tolerance.

Mechanical Properties and Stress-Strain Relationship of PVA-Fiber-Reinforced Engineered Geopolymer Composite.

In this study, seven Engineering Geopolymer Composite (EGC) groups with varying proportions were prepared. Rheological, compressive, flexural, and axial tensile tests of the EGC were conducted to study the effects of the water/binder ratio, the cement/sand ratio, and fiber type on its properties. Additionally, a uniaxial tension constitutive model was established. The results indicate that the EGC exhibits early strength characteristics, with the 7-day compressive strength reaching 80% to 92% of the 28-day compressive strength. The EGC demonstrates high compressive strength and tensile ductility, achieving up to 70 MPa and 4%, respectively. The mechanical properties of the EGC improved with an increase in the sand/binder ratio and decreased with an increase in the water/binder ratio. The stress-strain curve of the EGC resembles that of the ECC, displaying a strain-hardening state that can be divided into two stages: before cracking, the matrix primarily bears the stress; after cracking, the slope decreases, and the fiber predominantly bears the stress.

Self- and Situation-Focused Reappraisal are not homogeneous: Evidence from behavioral and brain networks.

Reappraisal is an effective emotion regulation strategy which can be divided into self- and situation-focused subtypes. Previous studies have produced inconsistent findings on the moderating effects and neural mechanisms of reappraisal; thus, further research is necessary to clarify these inconsistencies. In this study, a total of 44 participants were recruited and randomly assigned to two groups. 23 participants were assigned to the self-focused group, while 21 participants were assigned to the situation-focused group. The participants' resting EEG data were collected for 6 min before the experiment began, followed by an emotional regulation task. During this task, participants were asked to view emotion-provoking images under four emotion regulation conditions (View, Watch, Increase, or Decrease). Late positive potential (LPP) was obtained when these emotional images were observed. LPP is an effective physiological indicator of emotion regulation, enabling this study to explore emotion regulation under different reappraisal strategies, as well as the functional connectivity and node efficiency within the brain. It was found that, in terms of the effect on emotion regulation, situation-focused reappraisal was significantly better than self-focused reappraisal at enhancing the valence of negative emotion, while self-focused reappraisal was significantly better than situation-focused reappraisal at increasing the arousal of negative emotion. In terms of neural mechanisms, multiple brain regions such as the anterior cingulate cortex, the frontal lobe, the parahippocampal gyrus, parts of the temporal lobe, and parts of the parietal lobe were involved in both reappraisal processes. In addition, there were some differences in brain regions associated with different forms of cognitive reappraisal. Self-focused reappraisal was associated with the posterior cingulate gyrus, fusiform gyrus, and lingual gyrus, and situation-focused reappraisal was associated with the parietal lobule, anterior central gyrus, and angular gyrus. In conclusion, this research demonstrates that self- and situation-focused reappraisal are not homogenous in terms of their effects and neural mechanisms and clarifies the uncertainties over their regulatory effects. Different types of reappraisal activate different brain regions when used, and the functional connectivity or node efficiency of these brain regions seems to be a suitable indicator for assessing the effects of different types of reappraisal.

A gene that underwent adaptive evolution, LAC2 (LACCASE), in Populus euphratica improves drought tolerance by improving water transport capacity.

Drought severely limits plant development and growth; accordingly, plants have evolved strategies to prevent water loss and adapt to water deficit conditions. However, experimental cases that corroborate these evolutionary processes are limited. The LACCASEs (LACs) family is involved in various plant development and growth processes. Here, we performed an evolutionary analysis of LACs from Populus euphratica and characterized the functions of LACs in Arabidopsis and poplar. The results showed that in PeuLACs, multiple gene duplications led to apparent functional redundancy as the result of various selective pressures. Among them, PeuLAC2 underwent strong positive selection. Heterologous expression analyses showed that the overexpression of PeuLAC2 alters the xylem structure of plants, including thickening the secondary cell wall (SCW) and increasing the fiber cell length and stem tensile strength. Altogether, these changes improve the water transport capacity of plants. The analysis of the physiological experimental results showed that PeuLAC2-OE lines exhibited a stronger antioxidant response and greater drought tolerance than WT. Three genes screened by transcriptome analysis, NAC025, BG1, and UGT, that are associated with SCW synthesis and drought stress were all upregulated in the PeuLAC2-OE lines, implying that the overexpression of PeuLAC2 thickened the SCW, improved the water transport capacity of the plant, and further enhanced its drought tolerance. Our study highlights that genes that have undergone adaptive evolution may participate in the development of adaptive traits in P. euphratica and that PeuLAC2 could be a candidate gene for molecular genetic breeding in trees.

Random topology organization and decreased visual processing of internet addiction: Evidence from a minimum spanning tree analysis.

OBJECTIVES: Internet addiction (IA) has been associated with widespread brain alterations. Functional connectivity (FC) and network analysis results related to IA are inconsistent between studies, and how network hubs change is not known. The aim of this study was to evaluate functional and topological networks using an unbiased minimum spanning tree (MST) analysis on electroencephalography (EEG) data in IA and healthy control (HC) college students. METHODS: In this study, Young's internet addiction test was used as an IA severity measure. EEG recordings were obtained in IA (n = 30) and HC participants (n = 30), matched for age and sex, during rest. The phase lag index (PLI) and MST were applied to analyze FC and network topology. We expected to obtain evidence of underlying alterations in functional and topological networks related to IA. RESULTS: IA participants showed higher delta FC between left-side frontal and parieto-occipital areas compared to the HC group (p < 0.001), global MST measures revealed a more star-like network in IA participants in the upper alpha and beta bands, and the occipital brain region was relatively less important in the IA relative to the HC group in the lower band. The correlation results were consistent with the MST results: higher IA severity correlated with higher Max degree and kappa, and lower eccentricity and diameter. CONCLUSIONS: Functional networks of the IA group were characterized by increased FC, a more random organization, and a decrease of relative functional importance of the visual processing area. Taken together, these alterations can help us understand the influence of IA to brain mechanism.

Coexpression of PalbHLH1 and PalMYB90 Genes From Populus alba Enhances Pathogen Resistance in Poplar by Increasing the Flavonoid Content.

Secondary metabolites of the flavonoid pathway participate in plant defense, and bHLH and MYB transcription factors regulate the synthesis of these metabolites. Here, we define the regulatory mechanisms in response to pathogens. Two transcription factors from Populus alba var. pyramidalis, PalbHLH1 and PalMYB90, were overexpressed together in poplar, and transcriptome analysis revealed differences in response to pathogen infection. The transgenic plants showed elevated levels of several key flavonoid pathway components: total phenols, proanthocyanidins (PAs), and anthocyanins and intermediates quercetin and kaempferol. Furthermore, PalbHLH1 and PalMYB90 overexpression in poplar enhanced antioxidase activities and H2O2 release and also increased resistance to Botrytis cinerea and Dothiorella gregaria infection. Gene expression profile analysis showed most genes involved in the flavonoid biosynthesis pathway or antioxidant response to be upregulated in MYB90/bHLH1-OE poplar, but significant differential expression occurred in response to pathogen infection. Specifically, expression of PalF3H (flavanone 3-hydroxylase), PalDFR (dihydroflavonol 4-seductase), PalANS (anthocyanin synthase), and PalANR (anthocyanin reductase), which function in initial, middle, and final steps of anthocyanin and PA biosynthesis, respectively, was significantly upregulated in D. gregaria-infected MYB90/bHLH1-OE poplar. Our results highlight that PalbHLH1 and PalMYB90 function as transcriptional activators of flavonoid pathway secondary-metabolite synthesis genes, with differential mechanisms in response to bacterial or fungal infection.

Erratum to: Subtelomeric multiplex ligation-dependent probe amplification as a supplement for rapid prenatal detection of fetal chromosomal aberrations.

[This corrects the article DOI: 10.1186/s13039-014-0096-1.].

Prenatal Diagnosis of DNA Copy Number Variations by Genomic Single-Nucleotide Polymorphism Array in Fetuses with Congenital Heart Defects.

OBJECTIVES: To evaluate the usefulness of single-nucleotide polymorphism (SNP) array for prenatal genetic diagnosis of congenital heart defect (CHD), we used this approach to detect clinically significant copy number variants (CNVs) in fetuses with CHDs. METHODS: A HumanCytoSNP-12 array was used to detect genomic samples obtained from 39 fetuses that exhibited cardiovascular abnormalities on ultrasound and had a normal karyotype. The relationship between CNVs and CHDs was identified by using genotype-phenotype comparisons and searching of chromosomal databases. All clinically significant CNVs were confirmed by real-time PCR. RESULTS: CNVs were detected in 38/39 (97.4%) fetuses: variants of unknown significance were detected in 2/39 (5.1%), and clinically significant CNVs were identified in 7/39 (17.9%). In 3 of the 7 fetuses with clinically significant CNVs, 3 rare and previously undescribed CNVs were detected, and these CNVs encompassed the CHD candidate genes FLNA (Xq28 dup), BCOR (Xp11.4 dup), and RBL2 (16q12.2 del). CONCLUSION: Compared with conventional cytogenetic genomics, SNP array analysis provides significantly improved detection of submicroscopic genomic aberrations in pregnancies with CHDs. Based on these results, we propose that genomic SNP array is an effective method which could be used in the prenatal diagnostic test to assist genetic counseling for pregnancies with CHDs.

Spontaneous electrical activities at myofascial trigger points at different stages of recovery from injury in a rat model.

BACKGROUND: Spontaneous electrical activity (SEA) is a feature of myofascial trigger points (MTrPs), which can either be latent or active. However, SEA at different stages of recovery from MTrPs remains unclear. OBJECTIVE: To investigate the temporal changes in the nature of SEA after generation of MTrPs in a rat model. METHODS: 32 rats were divided into four groups: 24 rats were assigned to experimental groups (EGs), which underwent the MTrP modelling intervention and 8 were allocated to a control group (CG). All EG rats received a blunt strike to the left vastus medialis combined with eccentric exercise for 8 weeks. After modelling, the EG rats were subdivided into three groups with total recovery times of 4, 8 and 12 weeks (EG-4w, EG-8w and EG-12w, respectively). Taut bands (TBs) with and without the presence of active MTrPs were identified in the left hind limb muscles of all rats, verified by SEA and further examined with electromyography recordings. Myoelectrical signals were also categorised into one of five types. RESULTS: CG rats had fewer TBs than EG rats and EGs showed variable frequencies of SEA. SEA frequencies were higher in EG-4w than in EG-8w and EG-12w groups (240.57±72.9 vs 168.14±64.5 and 151.63±65.4, respectively, p<0.05) and were significantly greater in all EGs than in the CG (55.75±21.9). Relative to CG rats, amplitudes and durations of electrical potentials in the EG were only increased in the EG-8w and EG-12w groups. Types IV and V myoelectrical signals were never seen in latent MTrPs and type V signals did not occur in EG-4w rats. CONCLUSIONS: Increasing recovery periods following a MTrP modelling intervention in rats are characterised by different frequencies and amplitudes of SEA from TBs. TRIAL REGISTRATION NUMBER: 2014012.

Subtelomeric multiplex ligation-dependent probe amplification as a supplement for rapid prenatal detection of fetal chromosomal aberrations.

BACKGROUND: Pregnant women with high-risk indications are highly suspected of fetal chromosomal aberrations. To determine whether Multiplex Ligation-dependent Probe Amplification (MLPA) using subtelomeric probe mixes (P036-E2 and P070-B2) is a reliable method for rapid detection of fetal chromosomal aberrations. The subtelomeric MLPA probe mixes were used to evaluate 50 blood samples from healthy individuals. 168 amniocytes and 182 umbilical cord blood samples from high-risk fetuses were analyzed using the same subtelomeric MLPA probe sets. Karyotyping was also performed in all cases of high-risk pregnancies, and single nucleotide polymorphism array analysis was used to confirm submicroscopic and ambiguous results from MLPA/karyotyping. RESULTS: Subtelomeric MLPA analysis of normal samples showed normal result in all cases by use of P036-E2 probe mix, while P070-B2 probe mix gave normal results for all but one case. In one normal control case P070-B2 produced a duplicated signal of probe for 13q34. In the high-risk group, totally 44 chromosomal abnormalities were found by karyotyping and MLPA, including 23 aneuploidies and 21 rearrangements or mosaics. MLPA detected all 23 aneuploidies, 12 rearrangements and 1 mosaic. Importantly, MLPA revealed 4 chromosomal translocations, 2 small supernumerary marker chromosomes (sSMCs), and 3 subtelomeric imbalances that were not well characterized or not detectable by karyotyping. However, MLPA showed negetive results for the remaining 8 rearrangements or mosaics, including 3 low mosaic aneuploidies, 1 inherited sSMC, and 4 paracentric inversions. CONCLUSIONS: Results suggest that combined use of subtelomeric MLPA and karyotyping may be an alternative method for using karyotype analyses alone in rapid detection of aneuploidies, rearrangements, and sSMCs.

Myoelectrical activity and muscle morphology in a rat model of myofascial trigger points induced by blunt trauma to the vastus medialis.

OBJECTIVES: To explore myoelectrical activity and muscle morphology of myofascial trigger points (MTrPs) in an injury model of rats. METHODS: A total of 24 male SD rats were randomly divided into a control group (group A) and model group (group B). A blunt striking injury and eccentric exercise were applied to the vastus medialis (VM) of rats in group B for 8 weeks. Later, the palpable taut band (TB), local twitch response, myoelectrical activities and morphology in the two groups were examined. RESULTS: An average of 2.5 (30/12) palpable TBs were detected in the VM in group B compared with none in group A. The MTrPs had two types of abnormal potential. Their amplitudes were significantly higher than those in the control group (p<0.01) but their durations showed no significant differences. A series of reflex contractions appeared in groups A and B in response to external stimulation to the ear. Their amplitude and duration in group B were significantly lower than those in group A. A series of lower fibrillation potentials repeatedly occurred in model MTrPs in group B. The morphology of MTrPs showed abnormal muscle fibres with large round or ellipse shapes in cross-section and enlarged tapering shapes in longitudinal section. CONCLUSIONS: Active MTrPs can be provoked by repeated blunt injury. Active MTrPs are a group of muscle fibres with abnormal shapes and abnormal myoelectrical potentials. External stimulation provokes low-voltage responses in MTrPs, which is different from the response of normal muscle fibres.