The development of rapeseed with high resistance against the pathogen Sclerotinia sclerotiorum is impeded by the lack of effective resistance resources within host species. Unraveling the molecular basis of nonhost resistance (NHR) holds substantial value for resistance improvement in crops. In the present study, small RNA sequencing and transcriptome sequencing were carried out between rice (a nonhost species of S. sclerotiorum) and rapeseed during infection, revealing the involvement of rice miRNAs on translation-related processes in both rice and the pathogen. Specifically, rice-specific miRNAs with potential capability for cross-kingdom RNAi against S. sclerotiorum were explored, of which Os-miR169y was selected as a representative case to elucidate its role in resistance to S. sclerotiorum. The silence of Os-miR169y decreased the resistance level of rice to S. sclerotiorum, and heterologous expression of Os-miR169y in Arabidopsis and rapeseed significantly enhanced the host resistance. The dual-luciferase reporter assay indicates that Os-miR169y targets S. sclerotiorum 60S ribosomal protein L19 (SsRPL19). Overexpressing Os-miR169y (OE_ss-miR169y) and RNAi of SsRPL19 (RNAi_ss-RPL19) in S. sclerotiorum significantly impaired the growth and pathogenicity of the pathogen, while overexpressing SsRPL19 exhibited a contrast effect. Yeast-two-hybridization revealed an interlinking role of SsRPL19 with multiple large and small ribosomal subunits, indicating its important role in translation. Proteome sequencing detected a decreased amount of proteins in transformants OE_ss-miR169y and RNAi_ss-RPL19 and significant suppression on key metabolic pathways such as carbon and nitrogen metabolisms. Collectively, this study suggests that rice can secrete specific miRNAs to suppress genes essential for S. sclerotiorum, such as Os-miR169y, which targets and suppresses SsRPL19 and thus impairs protein synthesis in the pathogen. This study sheds light on the intrinsic mechanisms of rice NHR against S. sclerotiorum, and further demonstrates the potential of using nonhost-specific “pathogen-attacking” miRNAs in improving resistance in host species.

Nitrate not only serves as the primary nitrogen source for terrestrial plants but also serves as a critical signal in regulating plant growth and development. Understanding how plant responses to nitrate availability is essential for improving nitrogen use efficiency in crops. Herein, we demonstrated that the basic helix-loop-helix (bHLH) transcription factor TabHLH489 plays a crucial negative regulatory role in wheat nitrate signaling. Overexpressing TabHLH489 significantly reduced nitrate-promoted wheat growth and grain yield. Transcriptomic analysis showed that approximately 75% of nitrate-responsive genes were no longerregulated by nitrate in the TabHLH489 overexpression lines. TabHLH489 directly interacts with TaNLP7-3A, the wheat homolog protein of NIN-like protein 7 (NLP7), a central transcription factor in nitrate signaling. This interaction impairs TaNLP7-3A's ability to bind DNA, thereby inhibiting its transcriptional activity. Moreover, TabHLH489 induces the accumulation of reactive oxygen species (ROS) to reduce the nuclear localization of TaNLP7-3A, thereby diminishing its effectiveness in regulating the plant nitrogen response. These findings highlight the intricate regulatory mechanism by which TabHLH489 modulates TaNLP7-3A activity through direct interaction and ROS-mediated inhibition of nuclear localization. Our research highlights the critical roles of TabHLH489 and TaNLP7-3A in modulating nitrate signaling, providing new gene targets for developing wheat varieties with enhanced nitrogen use efficiency.

Circular RNAs (circRNAs), a type of head-to-tail closed RNA molecules, have been implicated in various aspects of plant development and stress responses through transcriptome sequencing; however, the precise functional roles of circRNAs in plants remain poorly understood. In this study, we identified a highly expressed circular RNA, circZmMED16, derived from exon 8 of the mediator complex subunit 16 (ZmMED16) across different maize (Zea mays L.) inbred lines using circRNA-seq analysis. This circRNA is predominantly expressed in maize tassels and functions in the cytoplasm. Overexpression of circZmMED16 resulted in increased expression of ZmMED16/AtMED16 and delayed flowering in both maize and Arabidopsis thaliana, compared with that in wild-type plants. In contrast, overexpression of the parent gene ZmMED16 did not alter the flowering time of transgenic plants in Arabidopsis, suggesting that circZmMED16 plays a specific role in regulating flowering, distinct from that of linear ZmMED16. To further understand the mechanisms underlying the regulation of flowering time by circZmMED16, we performed RNA pull-down, dual-luciferase, RNA interference (RNAi), and ribonuclease protection assays (RPA). These results indicate that circZmMED16 interacts with small subunit 1 of ADP-glucose pyrophosphorylase (APS1) mRNA in both maize and Arabidopsis. The knockdown of circZmMED16 increased the expression of ZmAPS1, whereas the overexpression of circZmMED16 led to the downregulation of ZmAPS1 RNA and protein. By affecting ZmAPS1 expression, circZmMED16 reduced ADP-glucose pyrophosphorylase (AGPase) activity and led to delayed flowering. These results revealed a novel regulatory mechanism for circRNAs in flowering time and shed light on their functional and regulatory roles in plants.

Casbene and neocembrene are casbene-type macrocyclic diterpenes; their derivatives play significant roles in plant defense and have pharmaceutical applications. We had previously characterized a casbene synthase, TERPENE SYNTHASE 28 (OsTPS28), in rice (Oryza sativa). However, the mechanism of neocembrene biosynthesis in rice remained unclear. In this study, we identified two genes of the TPS-a1 subfamily, OsTPS2 and OsTPS10, encoding a neocembrene synthase and sesquiterpene synthase, respectively, as supported by enzyme activity assays and determination of subcellular localization. Metabolic profiling of rice lines overexpressing either TPS confirmed the catalytic functions of OsTPS2 and OsTPS10, and suggested that OsTPS10 enhances resistance to rice bacterial blight. An evolutionary analysis revealed that OsTPS10 is conserved in monocots and first appeared in wild rice, whereas OsTPS2 and OsTPS28 sequentially evolved through gene duplication, transit peptide recruitment, and mutation of key amino acids such as H362R. In summary, this study not only deepens our understanding of the metabolic pathways and evolutionary history governing the biosynthesis of casbene-type diterpenoids in rice, representing parallel and divergent evolution within the gene family, and offers gene resources for the improvement of rice.

Investigations into the nitrogen-fixing symbiosis between legumes and rhizobia can yield innovative strategies for sustainable agriculture. Legume species of the Inverted Repeat-Lacking Clade (IRLC) and the Dalbergioids, can utilize nodule cysteine-rich (NCR) peptides, a diverse family of peptides characterized by four or six highly conserved cysteine residues, to communicate with their microbial symbionts. These peptides, many of which exhibit antimicrobial properties, induce profound differentiation of bacteroids (semi-autonomous forms of bacteria) within nodule cells. This terminal differentiation endows the bacteroids with the ability to fix nitrogen, at the expense of their reproductive capacity. Notably, a significant number of NCR peptides is expressed in the nodule fixation zone, where the bacteroids have already reached terminal differentiation. Recent discoveries, through forward genetics approaches, have revealed that the functions of NCR peptides extend beyond antimicrobial effects and the promotion of differentiation. They also play a critical role in sustaining the viability of terminally differentiated bacteroids within nodule cells. These findings underscore the multifaceted functions of NCR peptides and highlight the importance of these peptides in mediating communications between host cells and the terminally differentiated bacteroids.

Plant regeneration is the process during which differentiated tissues or cells can reverse or alter their developmental trajectory to repair damaged tissues or form new organs. In the plant regeneration process, the cell wall not only functions as a foundational barrier and scaffold supporting plant cells but also influences cell fates and identities. Cell wall remodeling involves the selective degradation of certain cell wall components or the integration of new components. Recently, accumulating evidence has underscored the importance of cell wall remodeling in plant regeneration. Wounding signals, transmitted by transcription factors, trigger the expressions of genes responsible for cell wall loosening, which is essential for tissue repair. In de novo organ regeneration and somatic embryogenesis, phytohormones orchestrate a transcriptional regulatory network to induce cell wall remodeling, which promotes cell fate reprogramming and organ formation. This review summarizes the effects of cell wall remodeling on various regenerative processes and provides novel insights into the future research of uncharacterized roles of cell wall in plant regeneration.

Global climate change challenges agricultural production, as extreme temperature fluctuations negatively affect crop growth and yield. Low temperature (LT) stress impedes photosynthesis, disrupts metabolic processes, and compromises the integrity of cell membranes, ultimately resulting in diminished yield and quality. Notably, many tropical or subtropical horticultural plants are particularly susceptible to LT stress. To address these challenges, it is imperative to understand the mechanisms underlying cold tolerance in horticultural crops. This review summarizes recent advances in the physiological and molecular mechanisms that enable horticultural crops to withstand LT stress, emphasizing discrepancies between horticultural crops and model systems. These mechanisms include C-repeat binding factor-dependent transcriptional regulation, post-translational modifications, epigenetic control, and metabolic regulation. Reactive oxygen species, plant hormones, and light signaling pathways are integrated into the cold response network. Furthermore, technical advances for improving cold tolerance are highlighted, including genetic improvement, the application of light-emitting diodes, the utility of novel plant growth regulators, and grafting. Finally, prospective directions for fundamental research and practical applications to boost cold tolerance are discussed.

Leaf senescence can be triggered by various abiotic stresses. Among these, heat stress emerges as a pivotal environmental factor, particularly in light of the predicted rise in global temperatures. However, the molecular mechanism underlying heat-induced leaf senescence remains largely unexplored. As a cool-season grass species, tall fescue (Festuca arundinacea) is an ideal and imperative material for investigating heat-induced leaf senescence because heat stress easily triggers leaf senescence to influence its forage yield and turf quality. Here, we investigated the role of FaNAC047 in heat-induced leaf senescence. Overexpression of FaNAC047 promoted heat-induced leaf senescence in transgenic tall fescue that was evidenced by a more seriously destructive photosystem and higher accumulation of reactive oxygen species (ROS), whereas knockdown of FaNAC047 delayed leaf senescence. Further protein-DNA interaction assays indicated that FaNAC047 directly activated the transcriptions of NON-YELLOW COLORING 1 (FaNYC1), NYC1-like (FaNOL), and STAY-GREEN (FaSGR) but directly inhibited Catalases 2 (FaCAT2) expression, thereby promoting chlorophyll degradation and ROS accumulation. Subsequently, protein-protein interaction assays revealed that FaNAC047 physically interacted with FaNAC058 to enhance its regulatory effect on FaNYC1, FaNOL, FaSGR, and FaCAT2. Additionally, FaNAC047 could transcriptionally activate FaNAC058 expression to form a regulatory cascade, driving senescence progression. Consistently, the knockdown of FaNAC058 significantly delayed heat-induced leaf senescence. Collectively, our results reveal that FaNAC047-FaNAC058 module coordinately mediates chlorophyll degradation and ROS production to positively regulate heat-induced leaf senescence. The findings illustrate the molecular network of heat-induced leaf senescence for breeding heat-resistant plants.

Excessive exposure to high light can lead to photoinhibition, which impairs photosynthetic efficiency and causes oxidative damage in plants, such as sunburn in grapevines. This study investigates the role of resveratrol (Res), a stilbenoid with antioxidant properties, in protecting plants from high light damage. We found that exposure to high light increased reactive oxygen species (ROS) accumulation and induced photoinhibition in grapevine leaves. In response, Res biosynthesis was upregulated, along with an increase in stilbene synthase (VvSTS) expression. Application of exogenous Res alleviated ROS accumulation and improved photosynthetic efficiency. Further analysis revealed that the VvHY5-VvBEE1 regulatory module plays a pivotal role in regulating VvSTS expression under high light conditions. Specifically, VvHY5 activated VvSTS expression, while VvBEE1 repressed it. Transgenic analysis showed that overexpression of VvHY5 enhanced Res production and photoprotection, whereas overexpression of VvBEE1 reduced Res levels and exacerbated light-induced damage. VvHY5 and VvBEE1 competed for binding to the VvSTS promoter, with brassinosteroids (BRs) modulating their interaction. Our findings reveal the interplay between light signaling and brassinosteroid pathways in regulating Res biosynthesis, providing insights for protecting grapevines from sunburn.

In C₃ plants, photorespiration is an energy expensive pathway that competes with photosynthetic CO₂ assimilation and releases CO₂ into the atmosphere, potentially reducing C₃ plant productivity by 20%-50%. Consequently, reducing the flux through photorespiration has been recognized as a major way to improve C₃ crop photosynthetic carbon fixation and productivity. While current research efforts in engineering photorespiration are mainly based on the modification of chloroplast glycolate metabolic steps, only limited studies have explored optimizations in other photorespiratory metabolic steps. Here, we engineered an imGS bypass within the rice mitochondria to bypass the photorespiratory glycine toward glycine betaine, thereby, improving the photosynthetic carbon fixation in rice. The imGS transgenic rice plants exhibited significant accumulation of glycine betaine, reduced photorespiration, and elevated photosynthesis and photosynthate levels. Additionally, the introduction of imGS bypass into rice leads to an increase in the number of branches and grains per panicle which may be related to cytokinin and gibberellin signaling pathways. Taken together, these results suggest diverting mitochondrial glycine from photorespiration toward glycine betaine synthesis can effectively enhance carbon fixation and panicle architecture in rice, offering a promising strategy for developing functional mitochondrial photorespiratory bypasses with the potential to enhance plant productivity.

Photosynthetic organisms have developed various light-harvesting antenna systems to capture light and transfer energy to reaction centers (RCs). Simultaneous utilization of the integral membrane light-harvesting antenna (LH complex) and the extrinsic antenna (chlorosomes) makes the phototrophic bacterium Chloroflexus (Cfx.) aurantiacus an ideal model for studying filamentous anoxygenic phototrophs (FAPs). Here, we determined the structure of a minimal RC-LH photocomplex from Cfx. aurantiacus J-10-fl (CaRC-LH) at 3.05-Å resolution. The CaRC-LH binds only to seven LH subunits, which form a crescent-shaped antenna surrounding the movable menaquinone-10 (Q_B) binding site of CaRC. In this complex with minimal LH units, an extra antenna is required to ensure sufficient light-gathering, providing a clear explanation for the presence of chlorosomes in Cfx. aurantiacus. More importantly, the semicircle of the antenna represents a novel RC-LH assembly pattern. Our structure provides a basis for understanding the existence of chlorosomes in Cfx. aurantiacus and the possible assembly pattern of RC-LH.

Diatoms rely on fucoxanthin chlorophyll a/c-binding proteins (FCPs) for light harvesting and energy quenching under marine environments. Here we report two cryo-electron microscopic structures of photosystem I (PSI) with either 13 or five fucoxanthin chlorophyll a/c-binding protein Is (FCPIs) at 2.78 and 3.20 Å resolutions from Thalassiosira pseudonana grown under high light (HL) conditions. Among them, five FCPIs are stably associated with the PSI core, these include Lhcr3, RedCAP, Lhcq8, Lhcf10, and FCP3. The eight additional Lhcr-type FCPIs are loosely associated with the PSI core and detached under the present purification conditions. The pigments of this centric diatom showed a higher proportion of chlorophylls a, diadinoxanthins, and diatoxanthins; some of the chlorophyll as and diadinoxanthins occupy the locations of fucoxanthins found in the huge PSI-FCPI from another centric diatom Chaetoceros gracilis grown under low-light conditions. These additional chlorophyll as may form more energy transfer pathways and additional diadinoxanthins may form more energy dissipation sites relying on the diadinoxanthin-diatoxanthin cycle. These results reveal the assembly mechanism of FCPIs and corresponding light-adaptive strategies of T. pseudonana PSI-FCPI, as well as the convergent evolution of the diatom PSI-FCPI structures.

RNA interference (RNAi)-based control technologies are gaining popularity as potential alternatives to synthetic fungicides in the ongoing effort to manage plant pathogenic fungi. Among these methods, spray-induced gene silencing (SIGS) emerges as particularly promising due to its convenience and feasibility for development. This approach is a new technology for plant disease management, in which double-stranded RNAs (dsRNAs) targeting essential or virulence genes are applied to plants or plant products and subsequently absorbed by plant pathogens, triggering a gene silencing effect and the inhibition of the infection process. Spray-induced gene silencing has demonstrated efficacy in laboratory settings against various fungal pathogens. However, as research progressed from the laboratory to the greenhouse and field environments, novel challenges arose, such as ensuring the stability of dsRNAs and their effective delivery to fungal targets. Here, we provide a practical guide to SIGS for the control of plant pathogenic fungi. This guide outlines the essential steps and considerations needed for designing and assessing dsRNA molecules. It also addresses key challenges inherent to SIGS, including delivery and stability of dsRNA molecules, and how nanoencapsulation of dsRNAs can aid in overcoming these obstacles. Additionally, the guide underscores existing knowledge gaps that warrant further research and aims to provide assistance to researchers, especially those new to the field, encouraging the advancement of SIGS for the control of a broad range of fungal pathogens.