Plant, Cell & Environment最新文献

Protoplast-based systems provide a powerful and versatile platform for exploring how plants sense and respond to their environment. By enabling the direct delivery of proteins, DNA, and RNA into plant cells after cell wall removal, this approach facilitates precise molecular dissection of signaling, stress adaptation, and gene regulation across both model species and economically important crops. In this review, we analyzed 1050 published articles and categorizing them by delivery methods, research focus, plant species, and tissue types. We further highlight recent advances, including the application of single-cell transcriptomics, which provides unprecedented resolution for dissecting cellular responses and offers deeper insights into the mechanisms underlying stress resilience. Importantly, protoplast regeneration is gaining renewed attention not only as a model system for studying cellular reprogramming but also as a practical platform for crop improvement. Applications of protoplast regeneration include protoplast fusion, which integrates nuclear and organellar DNA/genomes from divergent parents to accelerate breeding and enhance tolerance to both biotic and abiotic stresses. Another important application is CRISPR/Cas ribonucleoprotein (RNP)-based editing targeting stress-resilience-related genes. In asexually propagated or highly heterozygous perennial crops with limited sexual reproduction, protoplast-based RNP delivery offers a viable and regulation-compliant strategy. This approach may help address public concerns over transgenic technologies while enabling the rapid development of stress-tolerant cultivars.

The composition of the root microbiota plays a crucial role in plant responses to soil-borne pathogens. Nevertheless, the defence mechanisms underlying multi-resistant soybean cultivars ability to combat major root rot pathogens remain poorly understood. This study aimed to elucidate how a resistant soybean, Heinong 531 (HN531), mitigates root rot through root metabolite secretion, microbial interactions, and biochemical strategies. We analysed the rhizosphere microbial community, secretion of antifungal compounds, and soil enzyme activities in HN531. A "protective microbial consortium" was identified and its role in pathogen suppression was investigated. Our results indicate that this community is associated with an enrichment of beneficial microorganisms, enhanced plant defence capacity, and increased soil enzyme activity, correlating with a disease control efficacy of up to 70% against root rot. These interactions involve the secretion of antimicrobial compounds and partially reshape the rhizosphere microbial structure, forming a protective microecological barrier. Our findings provide novel molecular targets for disease-resistant soybean breeding and highlight potential microbial resources for sustainable agriculture.

Rice is a major crop in China with a strong tendency to accumulate cadmium (Cd), posing serious risks to grain safety. α-Ketoglutarate (AKG), a key tricarboxylic acid cycle intermediate, has known roles in abiotic stress responses, but its effects on Cd tolerance and accumulation in rice remain unexplored. Exogenous AKG (50 μmol/L) was applied to investigate the mechanism regulating Cd tolerance and accumulation under both hydroponic and soil conditions. Under hydroponic experiments, exogenous AKG significantly reduced Cd translocation by 56.8%-63.9%, and decreased shoot Cd accumulation by 54.9%-60.6%. It alleviated photoinhibition and oxidative damage by enhancing photosynthesis and antioxidant activities while lowering H₂O₂ and MDA. Mechanistically, multi-omics analyses showed AKG confers Cd tolerance and Cd restriction concentration dependently. Under moderate Cd stress, AKG predominantly enhances the ascorbate-glutathione cycle and flavonoid biosynthesis for antioxidative defence, while upregulating OsHMA3 and reinforcing the endodermal barrier to restrict Cd transport. Under severe Cd stress, AKG shifts to activating melatonin biosynthesis and further suppressing transporters like OsIRT1. Additionally, AKG reduced grain Cd by 40.7% and increased yield by 31.6% under soil conditions. These results demonstrated the effective role of AKG in integrated Cd mitigation and provide a novel strategy for safe rice production in Cd-contaminated soils.

Retrograde transport from endosomes to the trans-Golgi network (TGN) is essential for intracellular trafficking, yet its molecular mechanism remains poorly understood. In Fusarium graminearum, 10 Rab GTPases associated with the Golgi-associated retrograde protein (GARP) complex were identified through immunoprecipitation followed by mass spectrometry (IP-MS). Among these, only the deletion of FgRAB6 disrupted the proper localisation of the GARP complex to the TGN. FgRab6 directly interacts with the GARP subunit FgVps52 via a conserved Q73 residue, which is critical for fungal growth and pathogenicity. Notably, this Q73-dependent interaction is evolutionarily conserved across eukaryotic species. Upon GTP activation, FgRab6 recruits FgVps52 to the TGN, thereby facilitating the assembly of the GARP complex through the sequential recruitment of additional subunits, including FgVps51, FgVps53 and FgVps54. The fully assembled GARP complex subsequently recruits the retromer complex and ensures the precise localisation of the SNARE proteins FgSnc1, FgTlg1 and FgTlg2 at the endosomes and the TGN. Disruption of this pathway severely compromises fungal development and virulence. Collectively, these findings identify a FgRab6-GARP-retromer-coordinated vesicle trafficking pathway that mediates the retrograde transport of SNARE proteins, which is critical for the pathogenicity of F. graminearum. This work provides new mechanistic insights into vesicular transport and highlights potential targets for antifungal intervention.

Citrus Huanglongbing (HLB), caused by 'Candidatus Liberibacter asiaticus' (CaLas), is the most devastating disease affecting the global citrus industry. Here, we reported that the CaLas effector SDE70 promotes HLB pathogenicity by targeting the citrus ubiquitination pathway. Transgenic expression of SDE70 in Wanjincheng orange (Citrus sinensis Osbeck) accelerated early CaLas proliferation, aggravated HLB symptoms, and increased susceptibility to citrus canker induced by Xanthomonas citri subsp. citri (Xcc). These results demonstrate that SDE70 functions as a broad-spectrum suppressor of citrus immunity. Mechanistically, SDE70 physically interacts with CsRUB2, a citrus ubiquitin-related protein. Furthermore, CsRUB2 overexpression in Wanjincheng oranges reduced resistance to HLB but enhanced resistance to citrus canker. Both SDE70 and CsRUB2 elevated salicylic acid (SA) and hydrogen peroxide (H₂O₂) levels in transgenic plants while lowering methyl salicylate (MeSA) levels. CsRUB2 also decreased jasmonic acid (JA). In contrast to the suppressive effect of SDE70, CsRUB2 enhanced the transcription of citrus immunity genes. Transient expression assays further demonstrated that the SDE70-CsRUB2 interaction dysregulates citrus immunity by perturbing SA, MeSA, JA, and H₂O₂ signals. These findings provide a theoretical basis for understanding citrus-CaLas interactions and breeding citrus varieties with broad-spectrum resistance to both HLB and citrus canker.

Freeze-thaw injury is a major cause of winter mortality in woody plants; however, the molecular mechanisms linking freeze-thaw stress to DNA damage and repair remain poorly defined. Here, we investigated the physiological thresholds of freeze-thaw injury in poplar and identified key regulatory components that enhance cold tolerance through improved DNA damage repair. Field temperature monitoring and differential scanning calorimetry revealed an effective freeze-thaw threshold of approximately 12°C, beyond which cumulative intracellular damage occurs despite the absence of extreme low temperatures. Integrated lncRNA, miRNA and mRNA sequencing demonstrated coordinated regulation of a DNA replication gene, PySLD5, by two long non-coding RNAs (MSTRG.19225.8 and MSTRG.19233.11) and the microRNA ptc-miR6476a. Functional assays, including pull-down, dual-luciferase and structural modelling, validated direct interactions among these RNAs and PySLD5. Overexpression of PySLD5 conferred enhanced cold tolerance, reduced electrolyte leakage and lower DNA fragmentation after freeze-thaw stress, whereas knockout lines showed severe cold sensitivity, disease susceptibility and reduced survival. Comet assays confirmed that repeated freeze-thaw cycles caused cumulative DNA damage. Together, these findings support a DNA damage accumulation model in which coordinated RNA regulation of PySLD5 promotes DNA repair, stabilizes replication forks and enhances overwintering survival.