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Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) systems have been successfully used as efficient tools for genome editing in a variety of species. We used the CRISPR/Cas9 system to mutate the Gn1a (Os01g0197700), DEP1 (Os09g0441900), GS3 (Os03g0407400), and IPA1 (Os08g0509600) genes of rice cultivar Zhonghua 11, genes which have been reported to function as regulators of grain number, panicle architecture, grain size and plant architecture, respectively. Analysis of the phenotypes and frequencies of edited genes in the first generation of transformed plants (T0) showed that the CRISPR/Cas9 system was highly efficient in inducing targeted gene editing, with the desired genes being edited in 42.5% (Gn1a), 67.5% (DEP1), 57.5% (GS3), and 27.5% (IPA1) of the transformed plants. The T2 generation of the gn1a, dep1, and gs3 mutants featured enhanced grain number, dense erect panicles, and larger grain size, respectively. Furthermore, semi-dwarf, and grain with long awn, phenotypes were observed in dep1 and gs3 mutants, respectively. The ipa1 mutants showed two contrasting phenotypes, having either fewer tillers or more tillers, depending on the changes induced in the OsmiR156 target region. In addition, we found that mutants with deletions occurred more frequently than previous reports had indicated and that off-targeting had taken place in highly similar target sequences. These results proved that multiple regulators of important traits can be modified in a single cultivar by CRISPR/Cas9, and thus facilitate the dissection of complex gene regulatory networks in the same genomic background and the stacking of important traits in cultivated varieties.

To address the limitations of conventional conveyor-based systems in online detection and grading of orange soluble solids content (SSC), this study developed a novel gravity-driven detection device. Traditional systems are constrained by carrier-induced optical interference, complex mechanical structures, and large spatial requirements, limiting their application in small- and medium-sized enterprises. By introducing a gravity-driven paradigm, this research eliminates the need for fruit carriers and enables vertical spectral acquisition during gravitational descent, effectively overcoming carrier interference and spatial constraints. The integrated system comprises a synchronous-release feeding mechanism, a Vis/NIR detection module, and an intelligent grading unit. Through systematic optimization of disk rotation speed, integration time, and spot size, stable and efficient spectral acquisition was achieved, resulting in a throughput of one fruit per second. The optimized PLSR model, utilizing SG-SNV preprocessing and CARS feature selection, demonstrated excellent predictive performance, with an Rp2 of 0.8746 and an RMSEP of 0.3001 °Brix. External validation confirmed 96.6% prediction accuracy within a ±1.0 °Brix error range and an overall grading accuracy of 86.6%. This system offers a compact, cost-effective, and high-performance solution for real-time fruit quality inspection, with potential applications to various spherical fruits.

This research addresses the limitations of traditional assembly line equipment, which is costly and impractical for narrow terrains, as well as the challenges of portable devices in large-scale detection. We propose a non-destructive testing method for assessing the internal quality of Sanhua Plums using a free-fall approach that integrates near-infrared spectroscopy and images. Through analysis of models created from spectral data collected under optimal conditions (motor speed: 6.6 r/min, integration time: 14 ms, spot diameter: 20 mm), we processed near-infrared data from 120 plums. The spectral data underwent preprocessing with polynomial smoothing (SG) and Standard Normal Variate (SNV) calibration, followed by feature extraction using Competitive Adaptive Reweighted Sampling (CARS), resulting in a prediction model for soluble solid content with R2 of 0.8374 and RMSE of 0.5014. Simultaneously, a prediction model based solely on visual image data achieved an R2 of 0.3341 and RMSE of 1.0115. We developed a multi-source information fusion model that incorporated Z-score normalization, linear weighted fusion, and Partial Least Squares Regression (PLSR), resulting in an R2 of 0.8871 and RMSE of 0.4141 for the test set. This model outperformed individual spectroscopy and visual models, supporting the development of an automated non-destructive system for evaluating Sanhua Plum’s internal quality.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a vector of Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus Huanglongbing (HLB). We examine the effectiveness of spraying adhesive on new shoots of orange jasmine (Murraya paniculata) to trap ACP in laboratory and field conditions and for the monitoring of ACP population dynamics and directional preferences. After 36 h of observation, orange jasmine plants with new shoots, with and without adhesive, are significantly (p < 0.05) more attractive to ACP than plants without new shoots. In field trials, orange jasmine with new shoots attracted more ACP, particularly females, than plants without new shoots. A male-biased ACP sex ratio occurred in a near-natural population. Orange jasmine with new shoots coated with adhesive more effectively trapped ACP than yellow sticky traps, particularly during the winter and early spring, when ACP densities were low. ACP has a strong phototropic response, preferring to feed and rest in south- and east-facing positions. Adhesive trapping shows potential for attracting adult ACP, especially in citrus orchards during cooler seasons, when host trees lack new shoots, and it may be particularly effective in doing so in urban areas and unmanaged citrus refugia (the primary sources of ACP infestations for commercial groves).

The AP2/ERF transcription factors play crucial roles in plant growth, development and responses to biotic and abiotic stresses. A total of 119 AP2/ERF genes (JcAP2/ERFs) have been identified in the physic nut genome; they include 16 AP2, 4 RAV, 1 Soloist, and 98 ERF genes. Phylogenetic analysis indicated that physic nut AP2 genes could be divided into 3 subgroups, while ERF genes could be classed into 11 groups or 43 subgroups. The AP2/ERF genes are non-randomly distributed across the 11 linkage groups of the physic nut genome and retain many duplicates which arose from ancient duplication events. The expression patterns of several JcAP2/ERF duplicates in the physic nut showed differences among four tissues (root, stem, leaf, and seed), and 38 JcAP2/ERF genes responded to at least one abiotic stressor (drought, salinity, phosphate starvation, and nitrogen starvation) in leaves and/or roots according to analysis of digital gene expression tag data. The expression of JcERF011 was downregulated by salinity stress in physic nut roots. Overexpression of the JcERF011 gene in rice plants increased its sensitivity to salinity stress. The increased expression levels of several salt tolerance-related genes were impaired in the JcERF011-overexpressing plants under salinity stress.

Salt stress interferes with plant growth and production. Plants have evolved a series of molecular and morphological adaptations to cope with this abiotic stress, and overexpression of salt response genes reportedly enhances the productivity of various crops. However, little is known about the salt responsive genes in the energy plant physic nut (Jatropha curcas L.). Thus, excavate salt responsive genes in this plant are informative in uncovering the molecular mechanisms for the salt response in physic nut. We applied next-generation Illumina sequencing technology to analyze global gene expression profiles of physic nut plants (roots and leaves) 2 hours, 2 days and 7 days after the onset of salt stress. A total of 1,504 and 1,115 genes were significantly up and down-regulated in roots and leaves, respectively, under salt stress condition. Gene ontology (GO) analysis of physiological process revealed that, in the physic nut, many \"biological processes\" were affected by salt stress, particular those categories belong to \"metabolic process\", such as \"primary metabolism process\", \"cellular metabolism process\" and \"macromolecule metabolism process\". The gene expression profiles indicated that the associated genes were responsible for ABA and ethylene signaling, osmotic regulation, the reactive oxygen species scavenging system and the cell structure in physic nut. The major regulated genes detected in this transcriptomic data were related to trehalose synthesis and cell wall structure modification in roots, while related to raffinose synthesis and reactive oxygen scavenger in leaves. The current study shows a comprehensive gene expression profile of physic nut under salt stress. The differential expression genes detected in this study allows the underling the salt responsive mechanism in physic nut with the aim of improving its salt resistance in the future.

Physic nut ( L.) is highly tolerant of barren environments and a significant biofuel plant. To probe mechanisms of its tolerance mechanisms, we have analyzed genome-wide transcriptional profiles of 8-week-old physic nut seedlings subjected to Pi deficiency (P-) for 2 and 16 days, and Pi-sufficient conditions (P+) controls. We identified several phosphate transporters, purple acid phosphatases, and enzymes of membrane lipid metabolism among the 272 most differentially expressed genes. Genes of the miR399/PHO2 pathway ( , miR399, and members of the SPX family) showed alterations in expression. We also found that expression of several transcription factor genes was modulated by phosphate starvation stress in physic nut seedlings, including an AP2/ERF gene ( ), which was down-regulated in both root and leaf tissues under Pi-deprivation. In -overexpressing Arabidopsis lines both numbers and lengths of first-order lateral roots were dramatically reduced, but numbers of root hairs on the primary root tip were significantly elevated, under both P+ and P- conditions. Furthermore, the transgenic plants accumulated less anthocyanin but had similar Pi contents to wild-type plants under P-deficiency conditions. Expression levels of the tested genes related to anthocyanin biosynthesis and regulation, and genes induced by low phosphate, were significantly lower in shoots of transgenic lines than in wild-type plants under P-deficiency. Our data show that down-regulation of the gene might contribute to the regulation of root system architecture and both biosynthesis and accumulation of anthocyanins in aerial tissues of plants under low Pi conditions.

Background The ‘Gonggan’ mandarin, an elite local cultivar from Zhaoqing City, Guangdong Province, combines the qualities of mandarin and sweet orange. A leaf-variegated mutant enhances its ornamental and economic value, providing an excellent model for studying chloroplast development and photosynthetic pigment metabolism in citrus. Results We found that, in this variegated mutant, chloroplasts are severely deficient or absent in mesophyll cells. Physiological assessments revealed lower levels of chlorophyll, carotenoids, net photosynthetic rate ( Pn ), and stomatal conductance ( Gs ), alongside significantly higher non-photochemical quenching (NPQ) and the non-photochemical quenching coefficient ( qN ), reflecting increased photoprotective energy dissipation. To uncover the molecular basis of leaf variegation, high-quality genome assemblies and transcriptomes were generated for both the normal and variegated ‘Gonggan’ mandarin, enabling comparative multi-omics analysis. Key genes involved in chloroplast development, such as TOC159 , PDV2 , THA8 , and SIG5 , were downregulated in the variegated leaves. Similarly, structural genes linked to chlorophyll degradation, including CLH2 , SGR , NOL , and NYC1 , exhibited altered expression. Downregulation of transcription factors GLK, GNC, and GNC-LIKE (GNL), known regulators of chloroplast development and chlorophyll biosynthesis, was also observed. Conclusions These findings suggest that disrupted expression of critical genes impacts chloroplast development and pigment metabolism, causing the leaf variegation phenotype. Overall, this study lays a foundation for functional genomics research and potential germplasm improvement of ‘Gonggan’ mandarin, and provides new insights into the mechanisms driving color variation in citrus.

The plant microbiome plays important roles in plant growth and resistance, but its assembly and affecting factors have not been fully studied for most of the agricultural plants. In this study, the endophytic mycobiota of the leaves and roots and the rhizosphere soils of five pummelo varieties were profiled based on the amplicon sequencing of the fungal internal transcribed spacer (ITS). The fungal richness and diversity were significantly different among the compartments, but not among the pummelo varieties. The composition and structure of the endophytic mycobiota of the compartments were significantly different across all five pummelo varieties. These suggest that the variety effect is weaker than the compartment effect, but still significant in shaping the pummelo mycobiota. Specifically, the dominant leaf endophytic fungal taxa (e.g., Fusarium and Zasmidium), and the root selection of fungal genera from the rhizosphere soils, were significantly different among the varieties. And also, the variety effect is more significant in shaping the leaf endophytic mycobiota than those of the roots. Finally, the pummelo varieties also showed some consistent alterations on the endophytic mycobiota, such as the root enrichment of Exophiala species. Our study indicates that the endophytic mycobiota of pummelos is significantly and interactively affected by plant variety and compartment effects, and suggests some fungi of interest for further tests.

Exocarpium Citri Grandis (ECG) is a distinctive medicinal and edible product originating from southern China and is often covered with a layer of characteristic “white frost” (WF). This study investigated the composition, formation mechanism, microbial safety, and anti-inflammatory activity of the WF. Multi-technique analyses revealed that WF mainly consisted of crystalline naringin (~80% of total mass). Drying-induced shrinkage and rupture of oil glands on ECG suggested metabolite migration and surface crystallization as the key mechanisms for WF formation. Microbial profiling revealed no significant differences in fungal and bacterial communities between WF and non-frost (NF) samples, and none of eight common mycotoxins was detected, confirming its microbial safety. Brewing tests demonstrated that water boiling for 30 min achieved efficient extraction of naringin, with higher yields in WF samples than in NF samples. In RAW264.7 cells, both WF and NF extract significantly inhibited lipopolysaccharide-induced NO production as well as the secretion and transcription of TNF-α, IL-6, IL-1β, iNOS, and NF-κB, with WF extract showing a stronger effect. Overall, these findings indicate that WF originates from endogenous flavonoid crystallization rather than microbial contamination and enhances the anti-inflammatory activity. This study provides a scientific basis for quality evaluation, processing optimization, and standardization of ECG products.