Search Results (6,862)

Eurasian aspen (Populus tremula L.) is a tree species with recognised ecological and economic importance for both natural and plantation forests. For the fast cloning of selected aspen genotypes, the method of plant propagation through in vitro culture (micropropagation) is often recommended. The efficiency of this method is related to the use of shoot-inducing chemical growth regulators, among which cytokinins, a type of plant hormone, dominate. Although cytokinins can inhibit rooting, this effect is avoided by using cytokinin-free media. This study sought to identify concentrations and combinations of growth regulators that would stimulate one type of P. tremula organogenesis (either shoot or root formation) without inhibiting the other. The investigated growth regulators included cytokinin 6-benzylaminopurine (BAP), auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA), auxins indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), gibberellin biosynthesis inhibitor paclobutrazol (PBZ), and a gibberellin mixture (GA₄/₇). Both BAP and TIBA increased shoot number per P. tremula explant and decreased the number of adventitious roots, but TIBA, in contrast to BAP, did not inhibit lateral root formation. However, for the maintenance of both adventitious shoot and root formation above the control level, the combination of PBZ and GA_4/7 was shown to be especially promising. Full article

Diploid Vaccinium fuscatum is a wild blueberry species with a low chilling requirement, an evergreen growth habit, and soil adaptability to southeast US growing regions. Regardless of its potential to improve the abiotic and biotic resilience of cultivated blueberries, this species has rarely been used for blueberry breeding. One hurdle is the ploidy barrier between diploid V. fuscatum and tetraploid cultivated highbush blueberries. To overcome the ploidy barrier, vegetative shoots micro-propagated from one genotype of V. fuscatum, selected because it grew vigorously in vitro and two southern highbush cultivars, ‘Emerald’ and ‘Rebel,’ were treated with colchicine. While shoot regeneration was severely repressed in ‘Emerald’ and ‘Rebel,’ shoot production from the V. fuscatum clone was not compromised at either 500 µM or 5000 µM colchicine concentrations. Due to the high number of shoots produced in vitro via the V. fuscatum clone shoots of this clone that had an enlarged stem diameter in vitro were subjected to flow cytometer analysis to screen for induced polyploidy. Sixteen synthetic tetraploid V. fuscatum, one synthetic octoploid ‘Emerald,’ and three synthetic octoploid ‘Rebel’ were identified. Growth rates of the polyploid-induced mutants were reduced compared to their respective wildtype controls. The leaf width and length of synthetic tetraploid V. fuscatum and synthetic octoploid ‘Emerald’ was increased compared to the wildtypes, whereas the leaf width and length of synthetic octoploid ‘Rebel’ were reduced compared to the wildtype controls. Significant increases in stem thickness and stomata guard cell length were found in the polyploidy-induced mutant lines compared to the wildtypes. In the meantime, stomata density was reduced in the mutant lines. These morphological changes may improve drought tolerance and photosynthesis in these mutant lines. Synthetic tetraploid V. fuscatum can be used for interspecific hybridization with highbush blueberries to expand the genetic base of cultivated blueberries. Full article

Fritillaria unibracteata is a rare and endangered medicinal plant in the Liliaceae family, whose bulbs have been used in traditional Chinese traditional medicine for over 2000 years. The mevalonate (MVA) pathway is involved in the growth, development, response to environmental stress, and active ingredient production of plants; however, the functional characterization of MVA-pathway genes in the Liliaceae family remains poorly documented. In this study, an Acetyl-CoA C-acetyltransferase gene (FuAACT) was first cloned from F. unibracteata. It exhibited structural features of the thiolase family and showed the highest sequence identity with the Dioscorea cayenensis homolog. The K_m, V_max, and K_cat of the recombinant FuAACT were determined to be 3.035 ± 0.215 μM, 0.128 ± 0.0058 μmol/(min·mg), and 1.275 ± 0.0575 min⁻¹, respectively. The optimal catalytic conditions for FuAACT were ascertained to be 30 °C and pH 8.9. It was stable below 50 °C. His361 was confirmed to be a key amino acid residue to enzymatic catalysis by site-directed mutagenesis. Subsequent subcellular localization experiments demonstrated that FuAACT was localized in chloroplasts and cytoplasm. FuAACT-overexpressing transgenic Arabidopsis thaliana plants showed higher drought tolerance than wild-type plants. This phenotypic difference was corroborated by significant differences in seed germination rate, lateral root number, plant height, and leaf number (p < 0.05). Furthermore, the FuAACT transgenic plants resulted in the formation of a more developed fibrous root system. These results indicated that the FuAACT gene revealed substantial biological activity in vitro and in vivo, hopefully providing the basis for its further research and application in liliaceous ornamental and medicinal plants. Full article

TasA gene, encoding a functional amyloid protein critical for biofilm formation and antimicrobial activity, was cloned from the endophytic strain Bacillus amyloliquefaciens BS-3, isolated from rubber tree roots. This study identified the shortest functional TasA variant (483 bp, 160 aa) reported to date, featuring unique amino acid substitutions in conserved domains. Bioinformatics analysis predicted a signal peptide (1–27 aa) and transmembrane domain (7–29 aa), which were truncated to optimize heterologous expression. Two prokaryotic vectors (pET28a and pCZN1) were constructed, with pCZN1-TasA expressed solubly in Escherichia coli Arctic Express at 15 °C, while pET28a-TasA formed inclusion bodies at 37 °C. Purified recombinant TasA exhibited potent antifungal activity, achieving 98.6% ± 1.09 inhibition against Colletotrichum acutatum, 64.77% ± 1.34 against Alternaria heveae. Notably, TasA completely suppressed spore germination in C. acutatum and Oidium heveae Steinmannat 60 μg/mL. Structural analysis via AlphaFold3 revealed that truncation enhanced protein stability. These findings highlight BS-3-derived TasA as a promising biocontrol agent, providing molecular insights for developing protein-based biopesticides against rubber tree pathogens. Full article

Climate extremes threaten the resilience of Eucalyptus plantations, yet hybridization with drought-tolerant species may enhance stress tolerance. This study analyzed xylem anatomical and functional drought responses in commercial Eucalyptus grandis (GG) clones and hybrids: E. grandis × camaldulensis (GC), E. grandis × tereticornis (GT), and E. grandis × urophylla (GU1, GU2). We evaluated vessel traits (water transport), fibers (mechanical support), and wood density (D) in stems and branches. Theoretical stem hydraulic conductivity (k_Stheo), vessel lumen fraction (F), vessel composition (S), and associations with previous hydraulic and growth data were assessed. While general drought responses occurred, GC had the most distinct xylem profile. This may explain it having the highest performance in different irrigation conditions. Red gum hybrids (GC, GT) maintained k_Stheo under drought, with stable F and a narrower vessel size, especially in branches. Conversely, GG and GU2 reduced F and S; and stem k_Stheo declined for a similar F in these clones, indicating vascular reconfiguration aligning the stem with the branch xylem. Almost all clones increased D under drought in any organ, with the highest increase in red gum hybrids. These results reveal diverse anatomical adjustments to drought among clones, partially explaining their growth responses. Full article

Cotton fiber initiation determines the fiber yield, yet the genetic basis underlying lint and fuzz initiation has still not been fully uncovered. Here, map-based cloning was carried out to identify the fiberless mutant genes derived from a cross between Gossypium hirsutum acc. WT and a natural fiberless mutant, fbl_SHZ. The 12:3:1 segregation ratio in F₂ populations (including 1848 and 3100 individuals that were developed in 2016 and 2018, respectively) revealed dominant epistasis, with the fuzz gene exerting dominance over the lint gene. Genetic linkage analysis revealed that GhMYB25like_A12 controls fuzz fiber initiation, while both GhMYB25like_A12 and GhMYB25like_D12 regulate lint fiber development. Sequencing analyses showed that the fbl_SHZ mutant exhibited a K104M mutation in the R2R3 domain of GhMYB25like_A12 and a transposable element insertion in GhMYB25like_D12, leading to fiberless seeds. Knockout of GhMYB25like_A12 produced fuzzless seeds, knockout of GhMYB25like_D12 led to no obvious change in seeds, and knockout of both (GhMYB25like_A12&D12) resulted in fiberless seeds. The 12:3:1 ratio reappeared in the F₂ population developed from the GhMYB25like_A12&D12 mutated plants as female and Jin668 as the male, which further confirmed the genetic interaction observed in fbl_SHZ. RNA-seq analysis revealed that GhMYB25like regulates cotton fiber initiation through multiple pathways, especially fatty acid metabolism. This study elucidates the key genes and their genetic interaction mechanisms governing cotton fiber initiation, providing a theoretical foundation for genetic improvement of cotton fiber traits. Full article

The barbel chub (Squaliobarbus curriculus) exhibits remarkable resistance to grass carp reovirus (GCRV), a devastating pathogen in aquaculture. To reveal the molecular basis of this resistance, we investigated complement factor D (DF)—a rate-limiting serine protease governing alternative complement pathway activation. Molecular cloning revealed that the barbel chub DF (ScDF) gene encodes a 1251-bp cDNA sequence translating into a 250-amino acid protein. Crucially, bioinformatic characterization identified a unique N-glycosylation site at Asn¹³⁹ in ScDF, representing a structural divergence absent in grass carp (Ctenopharyngodon idella) DF (CiDF). While retaining a conserved Tryp_SPc domain harboring the catalytic triad (His⁶¹, Asp¹⁰⁹, and Ser²⁰⁴) and substrate-binding residues (Asp¹⁹⁸, Ser²¹⁹, and Gly²²¹), sequence and phylogenetic analyses confirmed ScDF’s evolutionary conservation, displaying 94.4% amino acid identity with CiDF and clustering within the Cyprinidae. Expression profiling revealed constitutive ScDF dominance in the liver, and secondary prominence was observed in the heart. Upon GCRV challenge in S. curriculus kidney (SCK) cells, ScDF transcription surged to a 438-fold increase versus uninfected controls at 6 h post-infection (hpi; p < 0.001)—significantly preceding the 168-hpi response peak documented for CiDF in grass carp. Functional validation showed that ScDF overexpression suppressed key viral capsid genes (VP2, VP5, and VP7) and upregulated the interferon regulator IRF9. Moreover, recombinant ScDF protein incubation induced interferon pathway genes and complement C3 expression. Collectively, ScDF’s rapid early induction (peaking at 6 hpi) and multi-pathway coordination may contribute to barbel chub’s GCRV resistance. These findings may provide molecular insights into the barbel chub’s high GCRV resistance compared to grass carp and novel perspectives for anti-GCRV breeding strategies in fish. Full article

Resveratrol (RSV), a polyphenol found in a variety of berries and wines, is known for its anti-inflammatory, anticancer, and antioxidant properties. It has been suggested that RSV may play a role in the regulation of metabolic disorders, including diabetes and insulin resistance. However, in recent years, it has been reported to completely inhibit Akt kinase function in liver cells. Akt is a central protein involved in the metabolic function of insulin and is regulated by the phosphatidylinositol-3-kinase (PI3K) pathway. In this study, we examined the effect of RSV on insulin-induced insulin receptor (IR) phosphorylation and proteins involved in the PI3K/Akt pathway in a hepatic cell model, clone 9 (C9), and in hepatoma cells, Hepa 1-6 (H1-6). In both cell lines, RSV inhibited tyrosine phosphorylation of IR and insulin-induced activation of Akt. We also evaluated the effect of RSV on the activation of protein tyrosine phosphatase 1B (PTP1B), which is associated with IR dephosphorylation, and found that RSV increased PTP1B-Tyr¹⁵² phosphorylation in a time- and concentration-dependent manner. Furthermore, we found that the protein kinase C (PKC) inhibitors BIM and Gö6976 prevented the inhibition of Akt phosphorylation by RSV and increased the phosphorylation of Ser/Thr residues in IR, suggesting that PKC is involved in the inhibition of the insulin pathway by RSV. Thus, classical PKC isoforms impair the PI3K/Akt pathway at the IR and GSK3 and GS downstream levels; however, IRS-Tyr⁶³² phosphorylation remains unaffected. These results suggest that RSV can lead to insulin resistance by activating PTP1B and PKC, consequently affecting glucose homeostasis in hepatic cells. Full article

Enhancing the specificity and applicability of PCR-based genome-walking methods is highly desirable. A new and universal genome-walking tool, called center degenerated walking-primer PCR (CDWP-PCR), is presented in this study. CDWP-PCR involves adopting a center degenerated walking primer (cdWP) in the secondary/tertiary round of amplification. This cdWP is generated by degenerating the seven central nucleotides of the normal walking primer (nWP) used in primary PCR to NNNNNNN (where N includes the bases A, T, C, and G). Clearly, a partially complementary structure is formed between the two primers. Accordingly, the primary CDWP-PCR non-target products defined by the nWP are diluted in secondary/tertiary CDWP-PCR, as these non-targets have difficulty in annealing with the cdWP; conversely, the primary target product can still be efficiently amplified. The working performance of the proposed CDWP-PCR is verified through cloning of the unknown flanks of three known genes. All the clear DNA bands in the tertiary CDWP-PCRs are confirmed to be correct, and the largest DNA band is 8.0 kb. Overall, CDWP-PCR can be considered as a reliable supplement to existing genome-walking methods. Full article

Molecular chaperones, especially heat shock proteins (HSPs) have vital functions in cells’ responses to stress. Here, we cloned and sequenced the complete complementary DNA encoding HSP90 (MjHSP90) from the shrimp Marsupenaeus japonicus. The MjHSP90 cDNA comprised 3162 bp, including a 2172 bp coding region encoding a 724 amino acid-protein (predicted molecular mass = 83.12 kDa). Homology and phylogenetic analyses showed that MjHSP90 was highly conserved and most homologous to Litopenaeus vannamei HSP90. MjHSP90 is expressed in all tested tissues, with high expression in gill tissue and the hepatopancreas. Cold stress significantly upregulated MjHSP90 expression in the gill and hepatopancreas (p < 0.05). Following RNA interference knockdown of MjHSP90, the cold stress-related death rate of the shrimp increased significantly, accompanied by significantly upregulated expression of apoptosis-related genes Mjcaspase-3 and Mjbcl-2 (p < 0.05) and an increase in the number of apoptotic cells. The results indicated that MjHSP90 might play a pivotal role in the shrimp’s immune response to cold stress. Full article

As voice cloning technology rapidly advances, the risk of personal voices being misused by malicious actors for fraud or other illegal activities has significantly increased, making the collection of speech data increasingly challenging. To address this issue, this study proposes a data augmentation method based on XText-to-Speech (XTTS) synthesis to tackle the challenges of small-sample, multi-class speech recognition, using profanity as a case study to achieve high-accuracy keyword recognition. Two models were therefore evaluated: a CNN model (Proposed-I) and a CNN-Transformer hybrid model (Proposed-II). Proposed-I leverages local feature extraction, improving accuracy on a real human speech (RHS) test set from 55.35% without augmentation to 80.36% with XTTS-enhanced data. Proposed-II integrates CNN’s local feature extraction with Transformer’s long-range dependency modeling, further boosting test set accuracy to 88.90% while reducing the parameter count by approximately 41%, significantly enhancing computational efficiency. Compared to a previously proposed incremental architecture, the Proposed-II model achieves an 8.49% higher accuracy while reducing parameters by about 98.81% and MACs by about 98.97%, demonstrating exceptional resource efficiency. By utilizing XTTS and public corpora to generate a novel keyword speech dataset, this study enhances sample diversity and reduces reliance on large-scale original speech data. Experimental analysis reveals that an optimal synthetic-to-real speech ratio of 1:5 significantly improves the overall system accuracy, effectively addressing data scarcity. Additionally, the Proposed-I and Proposed-II models achieve accuracies of 97.54% and 98.66%, respectively, in distinguishing real from synthetic speech, demonstrating their strong potential for speech security and anti-spoofing applications. Full article

Despite rapid advances in robot learning, the coordination of closed-loop manipulation in cluttered environments remains a challenging and relatively underexplored problem. We present a novel two-level hierarchical architecture for a depth vision-equipped robotic arm that integrates pushing, grasping, and high-level decision making. Central to our approach is a prioritised action–selection mechanism that facilitates efficient early-stage learning via behaviour cloning (BC), while enabling scalable exploration through reinforcement learning (RL). A high-level decision neural network (DNN) selects between grasping and pushing actions, and two low-level action neural networks (ANNs) execute the selected primitive. The DNN is trained with RL, while the ANNs follow a hybrid learning scheme combining BC and RL. Notably, we introduce an automated demonstration generator based on oriented bounding boxes, eliminating the need for manual data collection and enabling precise, reproducible BC training signals. We evaluate our method on a challenging manipulation task involving five closely packed cubic objects. Our system achieves a completion rate (CR) of 100%, an average grasping success (AGS) of 93.1% per completion, and only 7.8 average decisions taken for completion (DTC). Comparative analysis against three baselines—a grasping-only policy, a fixed grasp-then-push sequence, and a cloned demonstration policy—highlights the necessity of dynamic decision making and the efficiency of our hierarchical design. In particular, the baselines yield lower AGS (86.6%) and higher DTC (10.6 and 11.4) scores, underscoring the advantages of content-aware, closed-loop control. These results demonstrate that our architecture supports robust, adaptive manipulation and scalable learning, offering a promising direction for autonomous skill coordination in complex environments. Full article

Radio Frequency Identification (RFID) technology is widely applied in various scenarios, including logistics tracking, supply chain management, and target monitoring. In these contexts, the malicious cloning of legitimate tag information can lead to sensitive data leakage and disrupt the normal acquisition of tag information by readers, thereby threatening personal privacy and corporate security and incurring significant economic losses. Although some efforts have been made to detect cloning attacks, the presence of missing tags in RFID systems can obscure cloned ones, resulting in a significant reduction in identification efficiency and accuracy. To address these problems, we propose the block-based cloned tag identification (BCTI) protocol for identifying cloning attacks in the presence of missing tags. First, we introduce a block indicator to sort all tags systematically and design a block mechanism that enables tags to respond repeatedly within a block with minimal time overhead. Then, we design a superposition strategy to further reduce the number of verification times, thereby decreasing the execution overhead. Through an in-depth analysis of potential tag response patterns, we develop a precise method to identify cloning attacks and mitigate interference from missing tags in probabilistic cloning attack scenarios. Moreover, we perform parameter optimization of the BCTI protocol and validate its performance across diverse operational scenarios. Extensive simulation results demonstrate that the BCTI protocol meets the required identification reliability threshold and achieves an average improvement of $24.01\%$ in identification efficiency compared to state-of-the-art solutions. Full article

Cryotherapy could stimulate immune responses and induce abscopal effects. A novel technique was developed for treating spinal bone tumors involving the use of frozen tumor-containing autologous bone grafts for anterior spinal reconstruction following total en-bloc spondylectomy, with the aim of activating cryoimmunity. This study focused on analyzing changes in the T-cell receptor (TCR) repertoire after surgery to evaluate T-cell diversity. Blood samples were collected pre- and post-operatively, with subsequent RNA extraction and immunosequencing. Compared to pre-surgery samples, the diversity and abundance of the Complementarity-Determining Region 3, regions of the TCR α and β chains decreased, suggesting that more selective clones may have emerged and influenced immune responses. Through TCR repertoire analysis, this study demonstrated that transplantation of frozen tumor-containing autologous bone impacted the immune system. This study is expected to provide a foundation for developing treatments that may enhance immune activation. Full article

An increase in invasive group A streptococcal (iGAS) infections among children under 15 years of age was reported in several countries between late 2022 and early 2023. This retrospective study aims to describe the epidemiology and clinical features of iGAS infections in children in Cyprus during the same period. Medical records of patients under 16 years old admitted with iGAS infection to the Archbishop Makarios Hospital, the only tertiary paediatric referral centre in Cyprus, between 1 January 2021 and 30 June 2024, were reviewed. Twenty-two cases were identified, of which twenty were classified as confirmed and two as probable. Half of the cases occurred in children aged 0–4 years, and 59% were recorded between December 2022 and April 2023. Scarlet fever was diagnosed in six children, five of whom developed pneumonia and empyema. Streptococcal toxic shock syndrome (STSS) was observed in five patients, resulting in two deaths and one case requiring prolonged extracorporeal membrane oxygenation (ECMO). The overall case fatality rate was 9.1%. Emm 1, belonging to the M1UK clone, was the predominant strain (66.6%). The findings underscore the severity of iGAS infections, particularly in younger children, and highlight the importance of timely diagnosis, appropriate management and continued epidemiological surveillance. Full article