aBIOTECH最新文献

N⁶-methyladenosine (m⁶A) is the most prevalent internal modification in eukaryotic mRNAs and contributes to the post-transcriptional regulation of gene expression. In plants, m⁶A modulates RNA splicing, stability, and translation, thereby influencing developmental processes and responses to environmental stimuli. This review systematically examines current advances in the understanding of m⁶A regulation in plants. We begin with an overview of the m⁶A modification and its associated regulatory machinery, including the writers (methyltransferases), erasers (demethylases), and readers (m⁶A-binding proteins) components, and discuss their roles in orchestrating RNA metabolism and determining plant phenotypes. Subsequent sections focus on the functional implications of m⁶A in economically important crops, with evidence drawn from model systems such as Arabidopsis thaliana and key species including rice (Oryza sativa), tomato (Solanum lycopersicum), and strawberry (Fragaria vesca), where m⁶A modifications have been linked to traits such as yield, maturation, and aroma. Finally, we explore emerging biotechnological strategies that harness m⁶A-mediated regulatory pathways to enhance crop quality, such as overexpression of human FTO encoding an m⁶A demethylase, quantitative m⁶A profiling at single-base resolution, CRISPR/Cas13-targeted m⁶A regulation, the application of small-molecule inhibitors, and m⁶A-driven multi-omics integration. These strategies provide a comprehensive framework for understanding the multifaceted roles of m⁶A in plant biology and underscore the potential of this modification as a target for next-generation crop improvement.

Rice (Oryza sativa) yield is a complex trait determined by multiple factors, including grain number per panicle (GPP), which is mainly genetically regulated. In this study, we identified the E3 RING DOMAIN LIGASE (RGLG) OsRGLG6 as a key regulator of GPP in rice. A yeast two-hybrid screen revealed that OsRGLG6 interacts with OsOTUB1, an ovarian tumor domain-containing ubiquitin-specific protease involved in panicle development. Further experiments confirmed the interaction between OsRGLG6 and OsOTUB1 in vitro and in vivo. OsRGLG6, which contains a vWA domain and a RING domain, is localized in the cytoplasm and endoplasmic reticulum and is highly expressed specifically in rice panicles. In vitro and in vivo ubiquitination assays demonstrated that OsRGLG6 ubiquitinates OsOTUB1 and promotes its degradation via the 26S proteasome pathway. CRISPR/Cas9-generated osrglg6 mutants exhibited significant reductions in GPP, indicating a crucial role for OsRGLG6 in the regulation of grain number. Transcriptome analysis revealed that OsRGLG6 regulates multiple downstream genes involved in plant hormone signaling, nitrogen utilization, and stress response, and the OsRGLG6–OsOTUB1 module specifically mediated drought-stress responses in a simulated drought experiment. This study provides insight into the molecular mechanisms that regulate rice GPP and yield, highlighting the potential of OsRGLG6 as a target for improvement of rice productivity.

Extreme hot weather severely limits rice (Oryza sativa) production. Rice cultivars from regions with hot weather are a valuable resource for breeding heat-tolerant rice, but the mechanisms mediating heat tolerance in these cultivars are not fully understood. Here, we investigated heat-tolerance mechanisms in rice using the well-known heat-tolerant cultivar Nagina 22 (N22) and comparing it with the less heat-tolerant cultivar 93–11. Following heat shock (HS) treatment (45 °C for 3 or 24 h), the expression of JASMONATE ZIM-DOMAIN (JAZ) genes spiked during the early stages of HS responses in N22 but not 93–11 and genes related to jasmonic acid (JA) signaling were repressed in N22. Promoting JA signaling in N22 via pretreatment with methyl JA (MeJA) impaired the heat tolerance of N22, measured as survival after HS treatment of 45 °C for 30 h, followed by a 7-d recovery. Furthermore, the N22-specific activation of JAZ genes was associated with increased histone acetylation and decreased DNA methylation. Comparing N22 to 93–11, we propose that the DNA demethylation process, rather than the hypomethylation status per se, is likely associated with JAZ activation. In summary, we revealed epigenetic mechanisms that may contribute to the heat tolerance of N22 via the JA signaling pathway; our findings have implications for improving heat tolerance in rice and other crops.

The development of effective genetic transformation tools is crucial for advancing molecular breeding in peanut (Arachis hypogaea). In this study, we identified and characterized a Ubiquitin (UBQ) promoter from peanut and evaluated its potential utility in transgenic research. Using sequence similarity–based identification and transcriptome analysis, we selected a highly expressed UBQ gene, arahy.E356RC, designated AhUBQ4, from which we cloned a 973-bp fragment of the promoter region. To assess its activity, we used this AhUBQ4 promoter fragment to drive expression of the GUS and Ruby reporter genes in transient and stable expression assays in Nicotiana benthamiana and peanut tissues. Compared to the commonly used CaMV 35S promoter, the AhUBQ4 promoter had stable and high transcriptional activity across multiple tissues. Furthermore, we replaced the traditional 35S promoter with the AhUBQ4 promoter in a CRISPR/Cas9 system, enabling efficient gene editing in peanut. Using a peanut hairy root transformation system, we induced site-specific mutations in HY5-HOMOLOG, confirming stable Cas9 expression from the AhUBQ4 promoter for genome editing applications. Our findings highlight the potential of the AhUBQ4 promoter as a valuable genetic tool for improving transformation efficiency and gene expression stability in peanut, paving the way for enhanced functional genomics studies and molecular breeding efforts.

Protein post-translational modifications such as phosphorylation and S-nitrosylation regulate protein functions and cellular programs in eukaryotes. Moreover, extensive evidence suggests crosstalk between these modifications. However, we lack a comprehensive method for the simultaneous detection and analysis of multiple post-translational modifications. Here, we present an optimized workflow that identifies phosphorylation and S-nitrosylation sites using a novel phosphate affinity tag switch technique. Validation with model proteins and complex biological samples confirmed the high sensitivity, coverage, and reproducibility of this method. Applying this method to Arabidopsis thaliana seedlings revealed 12,552 phosphorylation sites and 6,108 S-nitrosylation sites, representing the largest single-study dataset of S-nitrosylation sites to date. This approach enhances our understanding of post-translational modification dynamics in plant signaling, stress responses, and metabolism.

Plant-pathogenic fungi significantly affect crop yield and quality. Understanding pathogenic mechanisms and reducing yield losses from plant diseases are therefore crucial for global food security. Epigenetics has become a central focus in fungal biology research, and recent refinements in high-throughput sequencing technologies have drawn attention to the role of histone methylation in fungal pathogenicity. Due to their diversity and complexity, histone methylations play crucial roles in epigenetic and transcriptional regulation. In this review, we summarize recent progress in understanding histone methylation in plant-pathogenic fungi and examine how these modifications influence fungal pathogenicity. Ultimately, we aim to offer insight for creating fungal disease control strategies through the lens of histone methylation.

Peanut (Arachis hypogaea) is a widely cultivated legume crop that can fix nitrogen by forming root nodules with compatible rhizobia. The initiation and formation of these nodules require complex molecular communication between legumes and rhizobia, involving the precise regulation of multiple legume genes. However, the mechanism underlying nodulation in peanuts remains poorly understood. In this study, we identified a gene associated with nodulation in peanuts, named Peanut unique gene for nodulation 1.1 (AhPUGN1.1). Multiple lines of evidence indicate that AhPUGN1.1 is primarily expressed in peanut nodules. Silencing or knocking out AhPUGN1.1 in peanut resulted in fewer nodules, as well as lower fresh weight and nitrogenase activity, while overexpressing AhPUGN1.1 significantly enhanced nodulation ability and nitrogenase activity. Modulating the expression of AhPUGN1.1 also influenced the expression levels of genes associated with the Nod factor signaling pathway and infection via crack entry. Comparative transcriptome analysis revealed that AhPUGN1.1 likely regulates peanut nodulation by affecting the expression of genes involved in the cytokinin and calcium signaling pathways. Our data thus show that AhPUGN1.1 acts as a crucial regulator promoting symbiotic nodulation in peanuts.

Agriculture has become one of the largest users of non-renewable resources in the world and contributes heavily to resource depletion, environmental pollution, and climate change. Solutions to these problems are in dire need and these can partially be found in the inter-organismal interactions surrounding the rhizosphere of our crops. The rhizosphere is a highly complex ecosystem, serving as a habitat for a diverse array of beneficial and pathogenic organisms. Here, we review how plants are performing a balancing act, in which they employ chemical communication—through the exudation of chemicals from their roots—to recruit beneficial organisms, while keeping at the same time, pathogenic ones at bay. These metabolites released by roots are incredibly chemically diverse. Among them, isoprenoids, one of the most diverse metabolite classes, containing many, highly bioactive, molecules, are the focus of this review. A better insight into the chemical communication occurring between the root, the soil, and micro-organisms, will allow harnessing of the beneficial relationships and suppression of the harmful ones. Further, this will enable us to establish knowledge-based changes in how we perform agriculture, how we use chemical inputs, how we should breed more resilient crops and can bring back resilience to our agricultural soils.

Plant architecture is a major factor affecting crop management and yield. The erect leaf phenotype is a key trait for improving light capture, reducing water loss, optimizing space utilization, and facilitating the chemical and biological control of arthropods and pathogens, especially those infesting/infecting abaxial leaf surfaces. This phenotype has been associated with Tiller Angle Control 1 (TAC1)-like genes across many herbaceous and tree species. Our previous genomic and genetic analyses of the erect leaf phenotype in tomato (Solanum lycopersicum) indicated that this trait is controlled by a semi-dominant locus, Erl, on chromosome 10. We discovered that this phenotype was in tight linkage with a candidate loss-of-function mutation in Solyc10g009320, an ortholog of TAC1-like genes. Therefore, editing this gene might confirm its function and enable the fine-tuned manipulation of aboveground tomato plant architecture. Here, we utilized a CRISPR/Cas9 genome editing system to confirm the complete genetic association of the erect leaf phenotype in tomato by knocking out Solyc10g009320 in tomato cultivar ‘Micro-Tom’. In addition, we analyzed the effects of editing this gene on the overall plant phenotype as well as physiological and agronomic performance. Editing Solyc10g009320 alleles in tomato lays the foundation for the large-scale generation of superior genotypes, paving the way for the development of elite cultivars with an erect leaf phenotype.

Varieties with a semi-dwarf compact plant architecture may increase yield per unit area in rapeseed (Brassica napus) by allowing high-density cultivation and mechanical harvesting while conferring lodging resistance. Mutation of ERECTA (ER), which encodes a receptor-like protein kinase, generates a compact and upright plant architecture in Arabidopsis thaliana; however, there have been no reports on the roles of the ER family (ERf) in B. napus. In this study, we used the CRISPR/Cas9 system to generate mutants in each of the two homoeologs of B. napus ERf members BnaER and ER-Like 1 (BnaERL1), and in the single BnaERL2 gene, resulting in the homozygous mutants BnaA09.er/BnaC08.er, BnaA06.erl1/BnaC03.erl1, and BnaA10.erl2. Under greenhouse conditions, BnaA09.er/BnaC08.er plants were shorter than the wild type, with a compact inflorescence and shorter siliques. In addition, BnaA09.er/BnaC08.er plants produced significantly more branches and total siliques than the wild type, with no significant changes in the number of ovules per silique or thousand-seed weight. Under field conditions, the BnaA09.er/BnaC08.er mutant plant showed a phenotype comparable to that under greenhouse conditions, but with a notable drop in thousand-seed weight. These results indicate that the BnaA09.er/BnaC08.er mutant offers a valuable germplasm resource for breeding rapeseed with ideal plant architecture.