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CRISPR diagnostics are effective but suffer from low signal transduction efficiency, limited sensitivity, and poor stability due to their reliance on the trans-cleavage of single-stranded nucleic acid fluorescent reporters. Here, we present CrisprAIE, which integrates CRISPR/Cas reactions with “one to more” aggregation-induced emission luminogen (AIEgen) lighting-up fluorescence generated by the trans-cleavage of Cas proteins to AIEgen-incorporated double-stranded DNA labeled with single-stranded nucleic acid linkers and Black Hole Quencher groups at both ends (Q-dsDNA/AIEgens-Q). CrisprAIE demonstrates superior performance in the clinical nucleic acid detection of norovirus and SARS-CoV-2 regardless of amplification. Moreover, the diagnostic potential of CrisprAIE is further enhanced by integrating it with spherical nucleic acid-modified AIEgens (SNA/AIEgens) and a portable cellphone-based readout device. The improved CrisprAIE system, utilizing Q-dsDNA/AIEgen-Q and SNA/AIEgen reporters, exhibits approximately 80- and 270-fold improvements in sensitivity, respectively, compared to conventional CRISPR-based diagnostics. We believe CrisprAIE can be readily extended as a universal signal generation strategy to significantly enhance the detection efficiency of almost all existing CRISPR-based diagnostics. Current CRISPR diagnostic approaches can be hampered by several limitations, including low signal transduction efficiency and limited sensitivity. Here, the authors present CrisprAIE, an approach based on CRISPR/Cas-mediated “one to more” lighting-up nucleic acid detection using aggregation-induced emission luminogens.

Abiotic stresses such as drought and heat increasingly threaten plant growth and ornamental quality, particularly in climate-sensitive floricultural crops. Arbuscular mycorrhizal fungi (AMF) are known to enhance plant resilience under such conditions, yet their role in lilies remains insufficiently explored. In this study, we used a two-tier experimental approach to evaluate AMF-mediated benefits in lilies. First, different AMF strains, namely Funneliformis mosseae (FM), Rhizophagus intraradices (RI), Rhizophagus irregularis (RIG), Claroideoglomus etunicatum (CE), Diversispora versiformis (DV), and a mixed consortium (MIX), were screened for growth-promoting effects in two Lilium species, Taiwan lily and Lilium cv. Sorbonne, under non-stress conditions. Second, a selected AMF–host combination from the screening was evaluated to improve tolerance to drought, heat, and combined drought + heat stress. Among the tested strains, DV and MIX showed the most consistent improvements across key growth traits and root colonization. In the stress experiment, stress treatments reduced growth and physiological performance, particularly under combined drought + heat. AMF inoculation enhanced plant performance by improving shoot and root biomass, improving root system architecture, and leading to a higher chlorophyll content, greater relative water content, and enhanced flower traits. Biochemical analyses further revealed that AMF mitigated stress-induced oxidative damage by reducing reactive oxygen species (ROS) accumulation, as shown by reduced O2•− and H2O2 staining. This reduction in oxidative stress was supported by increased activities of key antioxidant enzymes, indicating that AMF activate cellular defense mechanisms. These findings underscore the potential of AMF as a sustainable biotechnological tool for improving stress tolerance in lilies and enhancing floricultural productivity under climate-challenged environments.

Trehalose is a nonreducing disaccharide critical for cellular integrity and stress adaptation in plants, and its synthesis relies on trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Despite their established roles in abiotic stress responses across model plants, these gene families remain underexplored in ornamental species like Petunia hybrida. Here, TPS and TPP genes in two wild petunia progenitors, P. axillaris and P. inflata, underwent a genome-wide analysis, with 10 TPS and 8–9 TPP genes being identified in each species. According to phylogenetic analysis, petunia TPS proteins cluster into two clades, while TPP proteins were classified into three clades, showing closer evolutionary ties to tomato homologs. Cis-acting elements profiling identified hormone- and stress-responsive regulatory elements (e.g., ABRE, TC-rich repeats). Expression analysis under drought, heat, and salt stress revealed dynamic temporal regulation. For instance, PaTPS4/PaTPS9 were early responders (peak at 6 h) under drought and salt stress, while PaTPS8 exhibited sustained upregulation during salt treatment. Heat stress uniquely suppressed PaTPS1,2 and PaTPP1, contrasting with broad upregulation of other members. Notably, PaTPP3 displayed delayed induction under heat. These findings underscore the functional diversity within TPS/TPP families, with specific members governing stress-specific responses. This study provides a foundational resource for leveraging these genes to enhance stress resilience and ornamental value in petunia.

Many countries around the world feature raw fish in their cuisine, which is valued for its unique flavor. However, raw fish may be easily contaminated with foodborne pathogens including Listeria monocytogenes, Salmonella, Vibrio parahaemolyticus, and Staphylococcus aureus. Herein, a method was established that integrated a multiplex polymerase chain reaction (PCR) and high-resolution melting (HRM) curve assay for the simultaneous detection of these four foodborne pathogens. The target genes of the bacteria were amplified by PCR and subsequently analyzed using HRM. Differentiation was achieved based on the melting temperature (Tm) values of their respective amplicons. The detection limit of the PCR-HRM assay was 0.02–0.1 ng/µL. In addition, the Tm remained nearly constant across various concentrations of genomic DNA derived from the target bacteria. The assay demonstrated perfect specificity (8/8) and a sensitivity of 5/5 for L. monocytogenes, 2/2 for Salmonella, 3/3 for V. parahaemolyticus, and 3/3 for S. aureus. No significant interference occurred when genomic DNA from the four target bacteria was co-extracted with DNA from eight non-target strains. Furthermore, the assay offers advantages including operational simplicity, high efficiency, accurate results, reduced detection time, and lower costs, rendering it well-suited for food safety applications in the aquatic products processing industry.

Salmonella is a type of common foodborne pathogen of global concern, seriously endangering human health. In molecular biological detection of Salmonella, the method of amplifying DNA often faces the problem of aerosol pollution. In this study, a microfluidic chip was developed to integrate loop-mediated isothermal amplification (LAMP) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system to detect Salmonella. The LAMP reaction solution was initially injected into the chamber to amplify at 65 °C for 20 min; the CRISPR/Cas12a reaction solution was subsequently injected to mix with the amplicons for fluorescent signal production at 43 °C for 30 min. Then, the results can be confirmed by naked eyes under 495 nm light or by a fluorescence immunochromatographic reader. The detection limit of this method for Salmonella DNA was 118 pg/μL. The sensitivity and specificity of this method was 100%. Furthermore, this method was used to detect Salmonella after enrichment for 4 h in salmon and chicken samples spiked with 30 CFU/25 g, and was verified to have a stable detection capability in real samples. The microfluidic chip integrated with the LAMP and CRISPR/Cas12a system not only provides a possibility of highly sensitive endpoint fluorescent visual detection of a foodborne pathogen, but also greatly eliminates the risk of aerosol contamination.

Wintersweet (Chimonanthus praecox L.) is a traditional winter-flowering plant in China and a popular cut flower in winter. Its unique flowering characteristics under cold stress may involve the regulation of a large number of proteins. Protein post-translational modification is an important regulating type for the gene function. However, little is known about the post-translational modification in wintersweet. SUMOylation is an important post-translational modification in eukaryotes. Small ubiquitin-like modifier (SUMO) E3 ligases perform multiple functional regulatory activities in plants via SUMOylation. Here, we cloned and identified a SIZ/PIAS-type SUMO E3 ligase, CpSIZ1, from wintersweet. CpSIZ1 shared high similarity with other SIZ1 proteins. Quantitative real-time PCR (qRT-PCR) indicated that CpSIZ1 was expressed in all tissues tested, with the highest expression in flower wither period of stage 6, and followed by mature leaves except for different flower development stages. The ectopic expression of CpSIZ1 in Arabidopsis, including the CpSIZ1 overexpression in siz1-2 mutant (HB line) and CpSIZ1 overexpression in WT (OE line), not only promoted vegetative growth, delayed flowering and accelerated leaf senescence, but also improve the cold tolerance in Arabidopsis. Therefore, our studies have enrich the understanding of function of SIZ1 gene in woody plant, and provide a good foundation for further research on the post-translational modification regulation mechanism in this winter-flowering plant.

Botrytis elliptica , the causal agent of gray mold disease, poses a major threat to commercial Lilium production, limiting its ornamental value and yield. The molecular and metabolic regulation mechanisms of Lilium 's defense response to B. elliptica infection have not been completely elucidated. Here, we performed transcriptomic and metabolomic analyses of B. elliptica resistant Lilium oriental hybrid “Sorbonne” to understand the molecular basis of gray mold disease resistance in gray mold disease. A total of 115 differentially accumulated metabolites (DAMs) were detected by comparing the different temporal stages of pathogen infection. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed the differentially expressed genes (DEGs) and DAMs were enriched in the phenylpropanoid and flavonoid pathways at all stages of infection, demonstrating the prominence of these pathways in the defense response of “Sorbonne” to B. elliptica . Network analysis revealed high interconnectivity of the induced defense response. Furthermore, time-course analysis of the transcriptome and a weighted gene coexpression network analysis (WGCNA) led to the identification of a number of hub genes at different stages, revealing that jasmonic acid (JA), salicylic acid (SA), brassinolide (BR), and calcium ions (Ca 2+ ) play a crucial role in the response of “Sorbonne” to fungal infection. Our work provides a comprehensive perspective on the defense response of Lilium to B. elliptica infection, along with a potential transcriptional regulatory network underlying the defense response, thereby offering gene candidates for resistance breeding and metabolic engineering of Lilium .

Wintersweet (Chimonanthus praecox) is familiar as a garden plant and woody ornamental flower. On account of its unique flowering time and strong fragrance, it has a high ornamental and economic value. Despite a long history of human cultivation, our understanding of wintersweet genetics and molecular biology remains scant, reflecting a lack of basic genomic and transcriptomic data. In this study, we assembled three cDNA libraries, from three successive stages in flower development, designated as the flower bud with displayed petal, open flower and senescing flower stages. Using the Illumina RNA-Seq method, we obtained 21,412,928, 26,950,404, 24,912,954 qualified Illumina reads, respectively, for the three successive stages. The pooled reads from all three libraries were then assembled into 106,995 transcripts, 51,793 of which were annotated in the NCBI non-redundant protein database. Of these annotated sequences, 32,649 and 21,893 transcripts were assigned to gene ontology categories and clusters of orthologous groups, respectively. We could map 15,587 transcripts onto 312 pathways using the Kyoto Encyclopedia of Genes and Genomes pathway database. Based on these transcriptomic data, we obtained a large number of candidate genes that were differentially expressed at the open flower and senescing flower stages. An analysis of differentially expressed genes involved in plant hormone signal transduction pathways indicated that although flower opening and senescence may be independent of the ethylene signaling pathway in wintersweet, salicylic acid may be involved in the regulation of flower senescence. We also succeeded in isolating key genes of floral scent biosynthesis and proposed a biosynthetic pathway for monoterpenes and sesquiterpenes in wintersweet flowers, based on the annotated sequences. This comprehensive transcriptomic analysis presents fundamental information on the genes and pathways which are involved in flower development in wintersweet. And our data provided a useful database for further research of wintersweet and other Calycanthaceae family plants.

Lily is one of the world’s important ornamental flowers. Potted Asiatic lily is a further selected dwarf cultivar suitable for indoor or garden planting. However, there is a lack of relevant research on the cultivation adaptability of potted Asiatic lilies cultivars in the Chongqing region which in the southwest of China. This study selected five potted Asiatic lily cultivars, and the phenological period, stem and leaf characteristics, and flowering traits were assessed through statistical observation. The Asiatic lily ‘Tiny Ghost’ and ‘Tiny Double You’ are well-suited for both spring and autumn planting in Chongqing, while ‘Sugar Love’ and ‘Curitiba’ are best planted in the spring. The ‘Tiny Diamond’ is more appropriate for autumn planting due to its low tolerance to high temperature. The application of exogenous substances, including calcium chloride (CaCl2), potassium fulvic acid (PFA) and melatonin (MT), can mitigate the detrimental effects of high-temperature stress on ‘Tiny Diamond’ by regulating photosynthesis, antioxidant systems, and osmotic substance content. A comprehensive evaluation using the membership function showed that the effect of exogenous CaCl2 treatment is the best, followed by exogenous PFA treatment. CaCl2 acts as a positive regulator of heat stress tolerance in Asian lilies, with potential applications in Asian lily cultivation. This study provides reference for cultivation and application of Asian lily varieties in Chongqing region, and also laid the foundation for further research on the mechanism of exogenous substances alleviating heat stress in lilies.