Search Results (125)

During fruit ripening in Capsicum species, substantial amounts of carotenoids accumulate in the pericarp. While the carotenoid biosynthesis pathway in Capsicum species has been extensively investigated from various angles, the transcriptional regulation of genes encoding carotenoid biosynthetic enzymes remains less understood in this non-climacteric horticultural crop compared to tomato, a climacteric fruit. In the present study, we investigated the function of the NAM, ATAF1/2 or CUC2 81 (CaNAC81) transcription factor gene. This gene was selected through RNA-Seq co-expression analysis based on the correlation between expressed transcription factor gene profiles and those of carotenoid structural genes. To determine its role in regulating the expression of biosynthetic-related carotenogenic genes, we performed Virus-Induced Gene Silencing (VIGS) assays in the Serrano-type C. annuum ‘Tampiqueño 74’. Fruits from plants infected with a pTRV2:CaNAC81 construct (silenced fruits) exhibited altered carotenoid pigmentation accumulation, manifested as yellow-orange spots, in contrast to fruits from non-agroinfected controls (NTC) and fruits from plants infected with the empty TRV2 construct (red fruits). Quantitative real-time PCR (qPCR) assays confirmed decreased transcript levels of CaNAC81 in fruits displaying altered pigmentation, along with reduced transcription of the PSY gene, which encodes the carotenoid biosynthetic enzyme phytoene synthase (PSY). High-performance liquid chromatography (HPLC) analysis revealed a distinct carotenoid pigment accumulation pattern in fruits from plants showing silencing symptoms, characterized by low concentrations of capsanthin and zeaxanthin and trace amounts of capsorubin, compared to control plants (NTC). These findings suggest the involvement of CaNAC81 in the regulatory network of the carotenoid biosynthetic pathway in chili pepper fruits. Full article

Traditional rice varieties (TRVs), shaped by generations of adaptation to local soils and climates, are often seen as less competitive than modern rice varieties (MRVs) due to lower yields. As a result, the spread of MRVs has contributed to a global decline in TRVs. However, TRVs offer notable advantages, particularly in terms of sustainability and health benefits. In light of their gradual disappearance, this study aimed to compare the nutritional quality and consumer preferences for selected TRVs and MRVs cultivated in Kerala, India. We evaluated sensory attributes and physicochemical properties to assess their influence on consumer preference. Sensory rankings were analyzed using Kendall’s W test, while multiple linear regression was used to examine the relationship between consumer preference and various quality parameters. The study found that TRVs had significantly higher antioxidant levels, while MRVs had substantially higher protein contents. Sensory evaluations ranked TRVs more favorably, with grain appearance and taste being key drivers of preference. Physicochemical characteristics also significantly influenced consumer choice. To understand how these preferences influenced purchasing behavior, we conducted exit surveys in supermarkets and applied cluster and discriminant function analyses. The results indicated that both younger consumers and senior residents preferred TRVs in terms of purchase and consumption patterns. Full article

COVID-19 restricted the number of employees. Operational data showed that traditional methods of modeling heat consumption are not correct anymore. The aim is to model the energy demand of an office building during COVID-19 limitations and showcase improvements after a new controller or suggested alternatives are applied. After an actual heat consumption profile was simulated, energy conservation scenarios were considered: the usage of thermostatic radiator valves (TRVs); accounting impacts of solar radiation and wind; changing mass flow rates based on the indoor temperature; adopting an additional control, changing the temperature setpoint; introducing night and day setbacks. After implementing new design and operational methods, the overheating of indoor spaces was alleviated, and the average indoor temperature was reduced from 23.5 °C to 20.4 °C. The annual specific heat consumption decreased to 174 kWh/m² (20.2% lower). The methodology ensured thermal comfort and high energy-saving potential. If operating parameters were adjusted, the total saving effect in energy demand was 119.8 MWh, with an energy-saving rate of 19.8%. Employing TRV-related savings and considering thermal inertia provided more stable indoor temperatures and higher energy performance. The minimum saving effect corresponded to the optimal operation and ensuring the indoor environment by considering wind and the maximum one-to-night setbacks. The fluctuations in indoor temperature became smoother. Full article

CLAVATA3 (CLV3)/endosperm surrounding region (CLE) peptides have been reportedly involved in plant growth and development, as well as responses to abiotic stresses. However, the stress resilience of most CLE genes in cotton remains largely unknown. Here, induced expression pattern analysis showed that GhCLE25 was obviously responsive to osmotic and salt treatments, indicating that GhCLE25 was involved in abiotic stress tolerance. Furthermore, silencing GhCLE25 or the exogenous application of CLE25p effectively led to reduced and enhanced drought tolerance, respectively, as indicated by the activities of the plants’ POD, SOD, CAT, and MDA contents, as well as their height and fresh weight. We found that the knockdown of GhCLE25 promoted seedling growth and development, with a higher plant height and fresh weight in GhCLE25-silenced plants in comparison to control plants. In addition, a comparative transcriptome analysis of TRV:00 versus TRV:GhCLE25 and Mock versus CLE25p revealed that the CLE25-mediated signaling pathway is mainly involved in defense response and phytohormone signaling. Collectively, these findings indicate diverse roles of CLE25 in regulating plant growth and response to environmental stimuli and highlight the potential utilization of CLE25 to improve drought stress in modern agriculture via CLE25p spraying. Full article

Soybean (Glycine max L.) is a vital grain and oil crop, serving as a primary source of edible oil, plant-based protein, and livestock feed. Its production is crucial for ensuring global food security. However, soybean yields are severely impacted by various diseases, and the development of disease-resistant cultivars remains the most sustainable strategy for mitigating these losses. While stable genetic transformation is a common approach for studying gene function, virus-induced gene silencing (VIGS) offers a rapid and powerful alternative for functional genomics, enabling efficient screening of candidate genes. Nevertheless, the application of VIGS in soybean has been relatively limited. In this study, we established a tobacco rattle virus (TRV)-based VIGS system for soybean, utilizing Agrobacterium tumefaciens-mediated infection. The TRV vector was delivered through cotyledon nodes, facilitating systemic spread and effective silencing of endogenous genes. Our results demonstrate that this TRV–VIGS system efficiently silences target genes in soybean, inducing significant phenotypic changes with a silencing efficiency ranging from 65% to 95%. Key genes, including phytoene desaturase (GmPDS), the rust resistance gene GmRpp6907, and the defense-related gene GmRPT4, were successfully silenced, confirming the system’s robustness. This work establishes a highly efficient TRV–VIGS platform for rapid gene function validation in soybean, providing a valuable tool for future genetic and disease resistance research. Full article

Objective: To investigate the inter-method agreement between in-person consultations and video recordings when diagnosing BPPV. Methods: Two experienced vestibular clinicians (clinician A and B) evaluated patients for the presence and type of BPPV, using the TRV chair (Interacoustics, Middlefart, DK), at a tertiary referral center. During these in-person consultations, diagnostic maneuvers and eye movements were recorded, and a diagnosis was made. Both clinicians independently evaluated their cases again, during two video review sessions (Video Review 1 and Video Review 2). These sessions were conducted one month apart. Both clinicians were blinded to patient information and medical history during the analysis and did not have access to sound recordings. They were asked to provide a new diagnosis, based on the videos alone. Inter-method and intra-observer agreement for BPPV diagnoses between in-person consultations and video reviews were assessed using the percentage of agreement and Cohen’s kappa. An independent analysis of all patients’ eye movements was conducted to identify patterns that might have influenced agreement between in-person consultation diagnoses and the two video reviews by clinicians. Results: During the in-person consultations, each clinician evaluated 100 patients. Clinician A diagnosed BPPV in 40% of the cases, while clinician B diagnosed it in 19% of the cases. Considering the inter-method agreement, clinician A agreed on 81% (95% CI (73, 89)) and 77% (95% CI (69, 85)) of the cases with associated kappa coefficients of 0.67 (95% CI (0.55, 0.79)) and 0.63 (95% CI (0.51, 0.75)) between in-person consultations and Video Reviews 1 and 2, respectively. For clinician B, the percentages of agreement were, respectively, 86% (95% CI (79, 93)) and 84% (95% CI (77, 91)), with corresponding kappa coefficients of 0.55 (95% CI (0.36, 0.74)) and 0.51 (95% CI (0.32, 0.70)). As for the intra-observer agreement, clinician A achieved an intra-observer agreement of 84% (95% CI (77, 91)) with kappa = 0.74 (95% CI (0.63, 0.85)), while clinician B achieved a slightly higher intra-observer agreement of 90% (95% CI (84, 96)) with kappa = 0.67 (95% CI (0.51, 0.83)). Descriptive analysis of the eye movement revealed that both clinicians showed high diagnostic consistency for “no BPPV” in cases without provoked nystagmus (78/86, 91%) even when spontaneous nystagmus was present, and for posterior canal BPPV (37/78, 47%) when characteristic nystagmus was observed. However, disagreement was noted for horizontal canal BPPV (15 cases) and in scenarios with subjective BPPV (2 cases) or purely vertical nystagmus (11/31 cases, 35%). Conclusions: This study showed the feasibility of using video recordings when diagnosing BPPV. It demonstrates that BPPV might be reliably diagnosed in a telemedicine setting. However, careful consideration must be given to certain factors during the protocol’s design to improve the diagnostic process. Full article

This study addresses the growing inadequacies of traditional architectural concepts and techniques in stormwater management amid the increasing frequency of extreme weather events, particularly in densely built urban micro-spaces. To tackle these challenges, we propose an integrated theoretical and practical framework applied to a case study of a small-scale urban public space in Chang’an District, Shijiazhuang City, Hebei Province, covering an area of about 2.15 hectares in North China. The framework combines Best Management Practices Planning (BMP-P) with Low Impact Development Design (LID-D). The framework optimizes sub-catchment delineation, strategically locates drainage outlets, and configures network layouts to reduce runoff path lengths, thereby reducing total runoff volume, enhancing drainage capacity, and alleviating surface water accumulation, which, in turn, informs the parametric design of LID facilities. In the BMP-P phase, four source-control measures were developed based on runoff control and stormwater retention: adjusting terrain slopes, adding or removing curbs and facilities, redistributing infiltration areas, and adjusting drainage outlet and piping layouts. By shortening runoff paths and reducing potential waterlogging areas, these measures effectively reduced total runoff volume (Trv) by 31.5% to 35.7% and peak runoff volume (Prv) by 19.4% to 32.4%. Moreover, by remodeling the stormwater network with a different layout, larger pipe diameters, and substantially increased network capacity, the total discharge (Tdv) increased by 1.8% to 50.2%, and the peak discharge rate (Pdr) increased by 100% to 550%, thus minimizing surface flooding. In the LID-D phase, we developed a Grasshopper-based parametric design program for the layout and design of LID facilities. This approach significantly reduces interdisciplinary communication costs and enhances urban planning efficiency. By integrating BMP and LID strategies, the proposed framework offers a flexible, rapid, and efficient solution for achieving resilient stormwater management in the context of urban micro-renewal. Full article

Lettuce is a globally cultivated and consumed leafy crop. Here we developed an efficient tobacco rattle virus (TRV)-based guide RNA (gRNA) delivery system for CRISPR/Cas editing in the commercial lettuce cultivar ‘Noga’. Plants stably expressing Cas9 were inoculated with TRV vectors carrying gRNAs targeting five nutrient-associated genes. The system achieved an average editing efficiency of 48.7%, with up to 78.9% of regenerated plantlets showing independent mutations. This approach eliminates the need for antibiotic selection, simplifying tissue culture processes. The system supports diverse applications, including Cas12a editing and large-fragment deletions using dual gRNA sets. Targeting the fructan 1-exohydrolase 2 (1-FEH2) gene produced knockout lines with significant increases in prebiotic dietary fibre fructan content, up to 5.2-fold, and an average rise in the degree of polymerisation by 2.15 units compared with controls. Combining 1-FEH1 and 1-FEH2 knockouts did not further increase fructan levels, revealing 1-FEH2 as the predominant isozyme in lettuce. RT-qPCR analysis showed reduced expression of the upstream biosynthetic enzyme sucrose:sucrose 1-fructosyl transferase (1-SST), suggesting potential feedback inhibition in fructan metabolism. This TRV-based gene editing approach, utilised here to increase fructan content, could be applied to improve other valuable traits in lettuce, and may inspire similar systems to enhance nutritional content of crops. Full article

The genetic transformation of plants has provided fundamental insights into plant biology. However, the genetic transformation systems for most horticultural plants remain incomplete. Genome editing has significantly contributed to the improvement of crop traits, but it heavily relies on effective genetic transformation. Currently, reducing costs and improving the efficiency of genetic transformation are crucial for promoting the widespread application of genome editing in plants. Here, we review the advances in plant genetic transformation research, performing analysis of three methods for plant gene function analysis that bypass tissue culture: Agrobacterium rhizogenes-mediated root genetic transformation, developmental regulators (DRs)-mediated genetic transformation, and virus-mediated genome editing. We analyzed transformation efficiency in strawberry and citrus using the A. rhizogenes infiltration method, employing GFP to label different subcellular locations to investigate the morphology of microfilaments, nuclei, and peroxisomes in strawberry cells. Sequence analysis revealed that a series of developmental regulators critical for enhancing genetic transformation efficiency in specific species are highly conserved across different plant species. Additionally, we successfully edited the endogenous Pds gene in Cas9-overexpressing transgenic tobacco using TRV and CLBV containing the gRNA module. These three methods offer the benefits of being cost-effective and time-efficient, providing valuable technical insights for the application of plant genome editing. Full article

Opioids are the most effective option for severe pain. However, it is well documented that the side effects associated with prolonged opioid use significantly constrain dosage in the clinical setting. Recently, researchers have concentrated on the development of biased opioid receptor agonists that preferentially activate the G protein signaling pathway over β-arrestin signaling. This approach is based on the hypothesis that G protein signaling mediates analgesic effects, whereas β-arrestin signaling is implicated in adverse side effects. Although certain studies have demonstrated that the absence or inhibition of β-arrestin signaling can mitigate the incidence of side effects, recent research appears to challenge these earlier findings. In-depth investigations into biased signal transduction of opioid receptor agonists have been conducted, potentially offering novel insights for the development of biased opioid receptors. Consequently, this review elucidates the contradictory roles of β-arrestin signaling in the adverse reactions associated with opioid receptor activation. Furthermore, a comparative analysis was conducted to evaluate the efficacy of the classic G protein-biased agonists, TRV130 and PZM21, relative to the traditional non-biased agonist morphine. This review aims to inform the development of novel analgesic drugs that can optimize therapeutic efficacy and safety, while minimizing adverse reactions to the greatest extent possible. Full article

Plant viruses utilize a subset of host plasmodesmata-associated proteins to establish infection in plants. In the present study, we aimed to understand the role of two plant genes, one encoding a putative plasmodesma located protein (PDLP) and a homolog of soybean gene regulated by cold 2 protein (SRC2) during Cucumber mosaic virus (CMV) infection. Virus-induced gene silencing (VIGS) was used to silence PDLP and SRC2 genes in Nicotiana benthamiana and in two related solanaceous plants, N. tabacum and Capsicum chinense Jacq. (Bhut Jolokia). Up to 50% downregulation in the expression of the PDLP gene using the TRV2-PDLP VIGS construct was observed in N. benthamiana and N. tabacum while, using the same gene construct, 30% downregulation of the target mRNA was observed in C. chinense. Similarly, using the TRV2-SRC2 VIGS construct, a 60% downregulation of the SRC2 mRNA was observed in N. benthamiana, N. tabacum, and a 40% downregulation in C. chinense as confirmed by qRT-PCR analysis. Downregulation of the PDLP gene in N. benthamiana resulted in delayed symptom appearance up to 7–12 days post inoculation with reduced CMV accumulation compared to the control plants expressing TRV2-eGFP. In contrast, SRC2-silenced plants showed enhanced susceptibility to CMV infection compared to the control plants. Our data suggest that the PDLP gene might facilitate infection of CMV, thus being a susceptibility factor, while the SRC2 gene could play a role in resistance to CMV infection in N. benthamiana. Full article

Rose is one of the most popular ornamental plants worldwide. The double-flower trait, referring to flowers with extra petals, has been a key focus in rose breeding history. However, the genetic mechanisms regulating petal number in roses are still not fully understood. Here, we identified the CUP-SHAPED COTYLEDON 3 (RhCUC3) gene in the miniature rose (Rosa hybrida ‘Eclair’). The expression of RhCUC3 was high during the petal and stamen primordium differentiation stages but declined sharply during pistil primordium development. RhCUC3 belongs to the NAM/CUC3 subgroup of NAC transcription factors and is localized in the nucleus. The transcript level of RhCUC3 increased significantly with ABA and GA treatments and was inversely down-regulated with MeJA and 6-BA treatments. Silencing RhCUC3 using virus-induced gene silencing (VIGS) in rose ‘Eclair’ significantly decreased the number of petaloid stamens and normal petals while slightly increasing the number of stamens. Additionally, the expression of RhAG and RhAGL, two MADS-box genes associated with floral organ identity, was significantly higher in TRV-RhCUC3 compared to the TRV control. These findings suggest that RhCUC3 enhances stamen petaloidy and petal number, potentially by modulating the expression of RhAG and RhAGL, providing new insights into the function of NAC transcription factors in plants. Full article

MYB (myeloblastosis) is one of the most abundant transcription factors in plants which regulates various biological processes. The molecular characteristics and function of R2R3-MYB transcription factors in amaranth remain unclear. In this study, 73 R2R3-MYB members were identified from the amaranth genome database and we further analyzed their chromosome position, conserved motifs, physiological and biochemical features, collinearity relationships, gene structure, phylogeny and cis-acting element. Based on the phylogenetic and expression pattern analysis, 14 candidate R2R3-MYB genes might be involved in the betalain synthesis. Amongst the 14 candidate R2R3-MYB genes, the expression level of AtrMYB72 was higher in ‘Suxian No.1’ than ‘Suxian No.2’, and also higher in the red section than in the green section of the same leaf in Amaranthus. The overexpression vector pCambia1301-AtrMYB72-GUS and VIGS (virus-induced gene silencing) vector pTRV2- AtrMYB72 were transferred into leaves of ‘Suxian No.1’ via an Agrobacterium-mediated method. The results showed that AtrMYB72 overexpression could promote betalain synthesis. A yeast one-hybrid assay and dual luciferase reporter gene assay demonstrated that AtrMYB72 could bind to the AtrCYP76AD1 promoter to promote betalain synthesis. These results indicated that AtrMYB72 promoted betalain biosynthesis in amaranth by activating the AtrCYP76AD1 transcription. Our results could provide new insights into the betalain biosynthesis in amaranth. Full article

Lily (Lilium Asiatica Hybrida) is a globally known perennial herbaceous bulbous flower, popular for its large, colourful flowers and high economic and ornamental value. However, pollen generation is a severe issue that reduces the cosmetic value of lilies. In this study, the MADS transcription factor LiAG1 was isolated and identified from the Lilium Asiatica Hybrida ‘Ice Pink Queen’, a male-sterile variety obtained through several years of hybridisation in our laboratory. qRT-PCR revealed that LiAG1 expression was greater in lily anthers, especially during the half-opening stage. The transient expression in tobacco demonstrated that LiAG1 was located in the nucleus. In the ‘Ice Pink Queen’ lily, suppression of LiAG1 using TRV-VIGS (tobacco-rattle-virus-mediated virus-induced gene silencing) resulted in the disappearance of most of the tapetum layer and the absence of the microsporangia. Overexpression of LiAG1 in transgenic Arabidopsis and tobacco resulted in narrower and more involute leaves, plant dwarfing, earlier blooming, and better pollen viability. Overall, our results suggested that LiAG1 might play an important role in flower development, especially anther development, of Lilium Asiatica Hybrida ‘Ice Pink Queen’. Full article

Compared to wheeled vehicles, tracked robotic vehicles have less ground pressure, greater traction adhesion, and stronger climbing and obstacle crossing capabilities, making them suitable for agricultural production in hilly areas. Good steering performance directly relates to the mobility performance and operating efficiency of tracked robotic vehicles. Affected by the ground slope, the ground pressure distribution of the vehicle’s two tracks is uneven, leading to changes in its steering performance. Therefore, analyzing and researching the steering performance of a tracked robotic vehicle under sloped conditions is of great significance. This study establishes a slope steering model for tracked robotic vehicles based on a ground pressure model of the multi-peak varying amplitude cosine distribution and the shearing displacement relationship between the track and the ground, and analyzes the impact of vehicle structural parameters, road surface parameters, and steering parameters on steering performance. To verify the proposed theoretical model, multi-body dynamics software is utilized for simulation modeling and analysis. Turning tests on different slopes are conducted on a “soil–machine–crop” integrated experimental platform. The relative error between the numerical analysis results and the virtual simulation software’s results is less than 12%, and the relative error between the numerical analysis results and the experimental results is less than 10.3%; the good consistency between the theoretical results and the simulation’s results and the experimental results indicates that the model is, indeed, correct and effective. The established steering model can provide a theoretical basis for the design and control of new steering mechanisms for agricultural tracked robotic vehicles. Full article