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Abstract Huanglongbing (HLB), associated with the psyllid-vectored phloem-limited bacterium, Candidatus Liberibacter asiaticus (CLas), is a disease threat to all citrus production worldwide. Currently, there are no sustainable curative or prophylactic treatments available. In this study, we utilized mass spectrometry (MS)-based metabolomics in combination with 3D molecular mapping to visualize complex chemistries within plant tissues to explore how these chemistries change in vivo in HLB-infected trees. We demonstrate how spatial information from molecular maps of branches and single leaves yields insight into the biology not accessible otherwise. In particular, we found evidence that flavonoid biosynthesis is disrupted in HLB-infected trees, and an increase in the polyamine, feruloylputrescine, is highly correlated with an increase in disease severity. Based on mechanistic details revealed by these molecular maps, followed by metabolic modeling, we formulated and tested the hypothesis that CLas infection either directly or indirectly converts the precursor compound, ferulic acid, to feruloylputrescine to suppress the antimicrobial effects of ferulic acid and biosynthetically downstream flavonoids. Using in vitro bioassays, we demonstrated that ferulic acid and bioflavonoids are indeed highly bactericidal to CLas, with the activity on par with a reference antibiotic, oxytetracycline, recently approved for HLB management. We propose these compounds should be evaluated as therapeutics alternatives to the antibiotics for HLB treatment. Overall, the utilized 3D metabolic mapping approach provides a promising methodological framework to identify pathogen-specific inhibitory compounds in planta for potential prophylactic or therapeutic applications.

Abstract Long intergenic non-coding RNAs (lincRNAs) are emerging as regulators of protein-coding genes (PCGs) in many plant and animal developmental processes and stress responses. In this study, we characterize the genome-wide lincRNAs in potatoes responsive to a vascular bacterial disease presumably caused by Candidatus Liberibacter solanacearum (CLso). Approximately 4397 lincRNAs were detected in healthy and infected potato plants at various stages of zebra chip (ZC) disease progression. Of them, ~65% (2844) were novel lincRNAs, and less than 1% (9) were orthologs of Arabidopsis and rice based on reciprocal BLAST analysis, suggesting species-specific expansion. Among the proximal lincRNAs within 50 kbp from a PCG, ~49% were transcribed from the same strand, while ~39% and ~15% followed convergent (head-to-head) and divergent (tail-to-tail) orientations, respectively. Approximately 30% (1308) were differentially expressed following CLso infection, with substantial changes occurring 21 days after infection (DAI). Weighted Gene Co-expression Network Analysis (WGCNA) of lincRNAs and PCGs identified 46 highly correlated lincRNA–PCG pairs exhibiting co-up or co-downregulation. Furthermore, overexpression of selected lincRNAs in transgenic potato hairy roots resulted in perturbation of neighboring PCG expression and conferred tolerance to CLso infection. Our results provide novel insights into potato lincRNAs’ identity, expression dynamics, and functional relevance to CLso infection.

Green synthesis methods for making nanoparticles using plant extracts have gained considerable attention in recent years, particularly because of their potential applications in nanomedicine. Plant biosynthesized nanoparticles have shown noteworthy biomedical uses in comparison. The silver nanoparticle (AgNP) synthesized using extracts from the leaves and flowers of Cymbopogon citratus Linn was investigated for its antibacterial and anticancer properties. The biosynthesized AgNPs were characterized using UV–Vis, FTIR, DLS, EDX and SEM. The presence of alcohol, alkene, and phytochemicals in the AgNPs was confirmed using FTIR analysis. The investigation using SEM with EDX spectroscopy confirmed that the AgNPs were both pure and in the form of nanocrystals. Furthermore, AgNPs demonstrated significant antibacterial activity by efficiently suppressing bacterial growth. The biosynthesized AgNPs exhibited a concentration-dependent reduction in the lung cancer-A549 cell growth. The ethanolic extracts of C. citratus flower-synthesized AgNPs contain a wide range of phytochemical constituents. These components have been shown to effectively modulate antibacterial and anticancer activities, superior to the effects of leaf extracts synthesized AgNPs. Consequently, additional research is required to investigate the potential biomedical applications of these biosynthesized AgNPs.

Summary: “Candidatus Liberibacter spp.” are insect-vectored, fastidious, and vascular-limited phytopathogens. They are the presumptive causal agents of potato zebra chip, tomato vein clearing, and the devastating citrus greening disease worldwide. There is an urgent need to develop new strategies to control them. In this study, we characterized a dual-specificity serine/tyrosine phosphatase (STP) that is well conserved among thirty-three geographically diverse “Candidatus Liberibacter spp.” and strains that infect multiple Solanaceaea and citrus spp. The STP is expressed in infected plant tissues, localized at the plant cytosol and plasma membrane, and interferes with plant cell death responses. We employed an in silico target-based molecular modeling and ligand screen to identify two small molecules with high binding affinity to STP. Efficacy studies demonstrated that the two molecules can inhibit “Candidatus Liberibacter spp.” but not unrelated pathogens and confer plant disease tolerance. The inhibitors and strategies are promising means to control “Candidatus Liberibacter spp.”

Potato zebra chip (ZC) disease, associated with the uncultured phloem-limited bacterium, Candidatus Liberibacter solanacearum (CLso), is transmitted by the potato psyllid Bactericera cockerelli. Potato ZC disease poses a significant threat to potato production worldwide. Current management practices mainly rely on the control of the psyllid to limit the spread of CLso. The present study investigated new sources of ZC resistance among wild Solanum species. A taxonomically diverse collection of tuber-bearing Solanum species was screened; one ZC-resistant accession and three ZC-tolerant accessions were identified among the 52 screened accessions. Further characterization of the resistant accession showed that the resistance was primarily associated with antibiosis effects due to differences in leaf trichome density and morphology of the wild accession, which could limit the psyllid feeding and oviposition. This germplasm offers a good resource for further understanding ZC and psyllid resistance mechanisms, contributing to potato breeding efforts to develop ZC resistance cultivars. Alternatively, it could be used as a potential trap crop to manage psyllid and control ZC disease.

The putative causal agent of citrus greening Candidatus Liberibacter asiaticus (CLas) cannot be cultured, which hampers finding new therapies to control this devastating disease. Here, the authors show that hairy roots support CLas propagation and enable high throughput antimicrobial screening.

Sugarcane (Saccharum spp.) is a prominent source of sugar and serves as bioenergy/biomass feedstock globally. Multiple biotic and abiotic stresses, including drought, salinity, and cold, adversely affect sugarcane yield. G-protein-coupled receptors (GPCRs) are components of G-protein-mediated signaling affecting plant growth, development, and stress responses. Here, we identified a GPCR-like protein (ShGPCR1) from sugarcane and energy cane (Saccharum spp. hybrids) and characterized its function in conferring tolerance to multiple abiotic stresses. ShGPCR1 protein sequence contained nine predicted transmembrane (TM) domains connected by four extracellular and four intracellular loops, which could interact with various ligands and heterotrimeric G proteins in the cells. ShGPCR1 sequence displayed other signature features of a GPCR, such as a putative guanidine triphosphate (GTP)-binding domain, as well as multiple myristoylation and protein phosphorylation sites, presumably important for its biochemical function. Expression of ShGPCR1 was upregulated by drought, salinity, and cold stresses. Subcellular imaging and calcium (Ca2+) measurements revealed that ShGPCR1 predominantly localized to the plasma membrane and enhanced intracellular Ca2+ levels in response to GTP, respectively. Furthermore, constitutive overexpression of ShGPCR1 in sugarcane conferred tolerance to the three stressors. The stress-tolerance phenotype of the transgenic lines corresponded with activation of multiple drought-, salinity-, and cold-stress marker genes, such as Saccharum spp. LATE EMBRYOGENESIS ABUNDANT, DEHYDRIN, DROUGHT RESPONSIVE 4, GALACTINOL SYNTHASE, ETHYLENE RESPONSIVE FACTOR 3, SALT OVERLY SENSITIVE 1, VACUOLAR Na+/H+ ANTIPORTER 1, NAM/ATAF1/2/CUC2, COLD RESPONSIVE FACTOR 2, and ALCOHOL DEHYDROGENASE 3. We suggest that ShGPCR1 plays a key role in conferring tolerance to multiple abiotic stresses, and the engineered lines may be useful to enhance sugarcane production in marginal environments with fewer resources.

Citrus greening, also known as Huanglongbing (HLB), is caused by the unculturable bacterium Candidatus Liberibacter spp. (e.g., CLas), and has caused a devastating decline in citrus production in many areas of the world. As of yet, there are no definitive treatments for controlling the disease. Antimicrobial peptides (AMPs) that have the potential to block secretion-dependent effector proteins at the outer-membrane domains were screened in silico. Predictions of drug-receptor interactions were built using multiple in silico techniques, including molecular docking analysis, molecular dynamics, molecular mechanics generalized Born surface area analysis, and principal component analysis. The efflux pump TolC of the Type 1 secretion system interacted with natural bacteriocin plantaricin JLA-9, blocking the β barrel. The trajectory-based principal component analysis revealed the possible binding mechanism of the peptides. Furthermore, in vitro assays using two closely related culturable surrogates of CLas (Liberibacter crescens and Rhizobium spp.) showed that Plantaricin JLA-9 and two other screened AMPs inhibited bacterial growth and caused mortality. The findings contribute to designing effective therapies to manage plant diseases associated with Candidatus Liberibacter spp.

Potato (Solanum tuberosum L.) is an important food crop worldwide. As the demand for fresh and processed potato products is increasing globally, there is a need to manage and control devastating diseases such as zebra chip (ZC). ZC disease causes major yield losses in many potato-growing regions and is associated with the fastidious, phloem-limited bacterium Candidatus Liberibacter solanacearum (CLso) that is vectored by the potato-tomato psyllid (Bactericera cockerelli Šulc). Current management measures for ZC disease mainly focus on chemical control and integrated pest management strategies of the psyllid vector to limit the spread of CLso, however, they add to the costs of potato production. Identification and deployment of CLso and/or the psyllid resistant cultivars, in combination with integrated pest management, may provide a sustainable long-term strategy to control ZC. In this review, we provide a brief overview of the ZC disease, epidemiology, current management strategies, and potential new approaches to manage ZC disease in the future.

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) system is widely used as a genome-editing tool in various organisms, including plants, to elucidate the fundamental understanding of gene function, disease diagnostics, and crop improvement. Among the CRISPR/Cas systems, Cas9 is one of the widely used nucleases for DNA modifications, but manipulation of RNA at the post-transcriptional level is limited. The recently identified type VI CRISPR/Cas systems provide a platform for precise RNA manipulation without permanent changes to the genome. Several studies reported efficient application of Cas13 in RNA studies, such as viral interference, RNA knockdown, and RNA detection in various organisms. Cas13 was also used to produce virus resistance in plants, as most plant viruses are RNA viruses. However, the application of CRISPR/Cas13 to studies of plant RNA biology is still in its infancy. This review discusses the current and prospective applications of CRISPR/Cas13-based RNA editing technologies in plants.

Automation has been growing in recent years for the manufacturing industries to increase productivity. Multiple robotic arms are used to handle materials for lifting in flexible directions. The vertical rotation of a 360 degree single arm is considered in this research on a position servo drive with brushless DC motor. The load torque of an arm varies depending upon the angular displacement due to gravity, so it requires four-quadrant operation of the drive with a robust feedback controller. This paper deals with the design and performance comparison of a conventional PID feedback controller with a fuzzy-based PID controller and suggests the most suitable controller. The design was implemented in real time through the dSPACE DS1104 controller environment to verify the dynamic behaviors of the arm.

Aim: To evaluate and compare the variations in the inclination of occlusal plane of casts mounted on Artex articulator using a facebow with a fixed value and customized nasion indicator. Materials and Methods: Twenty two subjects were selected for this investigation. Two maxillary impressions were made, and casts poured. For each of the twenty two subjects, the facebow records were made with, Artex face-bow using a fixed value nasion indicator and customized nasion indicator and mounted. The angle between the occlusal plane of upper cast and the upper articulator arm was measured with a Universal bevel protractor and compared with the gold standard cephalometric value. Results: It shows that, when angle was measured between maxillary occlusal plane and upper member of the articulator, on the mounted cast using a customized nasion indicator and fixed value nasion indicator against the gold standard cephalometric value as a whole, it was found to be not significant. But, if each patient were evaluated individually, there found to be the difference in the angle. Discussion and Conclusion: Variation in occlusal plane was very minimal and close to the cephalometric value when using customized nasion indicator compared to fixed value nasion indicator on the Artex.

Advent of osseointegration has rapidly led to use of dental implants over recent years. Implant complications are often inadvertent sequelae of improper diagnosis, treatment planning, surgical method, and placement. This can be overcome by using surgical guides for implant positioning. Although conventionally made surgical guide are used, the clinical outcome is often unpredictable, and even if the implants are well placed, the location and deviation of the implants may not meet the optimal prosthodontic requirements. High accuracy in planning and execution of surgical procedures is important in securing a high success rate without causing iatrogenic damage. This can be achieved by computed tomography, 3D implant planning software, image-guided template production techniques, and computer-aided surgery. This article evaluates about the various systems of conventionally made surgical guide using radiograph and also the newer computer generated surgical guide in detail.

Background: The study was to identify the effects of ethanolic extract of Peltophorum pterocarpum leaves and bark on lipid profile, liver and carbohydrate enzymes on isoniazid and rifampicin induced Wistar rats. Materials and Methods: Ethanolic leaf and bark extract of P. pterocarpum freshly suspended in sterile water (300mg/kg BW), were administered to rats post-orally as a single dosage orally by intubation early morning for each day of the experimental period. Rats were separated into 10 groups with 6 rats in each group and maintained in isolated cages with proper ventilation. The samples were introduced to find the following parameters Total Cholesterol (TC), Triglycerides (TG), Low-Density Lipoprotein (LDL) cholesterol, High-Density Lipoprotein (HDL) cholesterol, Very-Low-Density Lipoprotein (VLDL) cholesterol, Phospholipids (PL), Free Fatty Acid (FFA), Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), Alkaline Phosphatase (ALP), glucose, glucose-6-phosphatase, glucose-6-phosphate dehydrogenase and hexokinase, kidney markers such as urea, uric acid and creatinine. Results: The results highlighted that the leaves and bark extract of P. pterocarpum had a potential hepatoprotective effect against the toxin treated rats and may alter lipid profile, liver and carbohydrate metabolizing enzymes levels. Conclusion: This effect proves the hepatoprotective nature of both the leaves and barks of P. pterocarpum. [ABSTRACT FROM AUTHOR]