Taraxacum kok-saghyz Rodin (TKS) is a promising alternative crop source for producing high-quality natural rubber (NR) and has become an ideal model plant for studying NR biosynthesis, regulation mechanisms, and production. So far, only a very limited number of functional genes related to NR biosynthesis have been identified in TKS. To achieve a systematic identification of its novel functional genes, we developed a mutant system denoted sense/antisense RNA expression (SARE) and have generated more than 8,000 transgenic TKS plants. A series of mutants with altered phenotypes, particularly changes in NR contents, were identified. To evaluate the efficiency of this library, we chose one mutant, c112, which exhibits a significant increase in NR content, for in-depth characterization. The c112 mutant arose from the sense insertion of a dormancy-associated gene1 (DRM1)/auxin repressed protein (ARP) gene, which we named high natural rubber content1 (HRC1). In the c112 mutant, the concentrations of NR precursors isopentenyl pyrophosphate and dimethylallyl diphosphate decreased, while geranylgeranyl diphosphate increased, suggesting that HRC1 regulates metabolic flux in NR biosynthesis. In summary, the developed TKS SARE mutant library provides valuable genetic resources for identifying key functional genes to accelerate the domestication of TKS from wild species to economic crops through molecular breeding.

Transcription factors (TFs) function as master regulators in multiple signaling pathways and govern diverse developmental and adaptive processes in plants. Some TFs identified in crop plants play critical roles in regulating yield through changes in plant architecture, including roots, stems, leaves, flowers, fruits, and grains. Although altering crop architecture can increase yields, the extent of yield enhancement is frequently hampered by diseases. Developing new crop varieties with improved yields and enhanced disease resistance remains challenging because immune system activation often impairs plant growth. Recently, approaches using TF engineering have made substantial progress in elevating both growth performance and disease resistance. However, most of these techniques rely on traditional transgenic methods. This review highlights discoveries in the last decade demonstrating improvements in growth performance, yield and immunity through TF engineering. We focus mainly on changes in plant architecture related to improved yield and disease resistance. We conclude with perspectives on the potential application of these discoveries for generating desirable crop traits by merging the most noteworthy biotechnology approaches, such as clustered regularly interspaced small palindromic repeats (CRISPR)/CRISPR-associated protein 9-mediated genome editing, with canonical molecular biology.

In plants, genetically encoded probes based on redox-sensitive green fluorescent protein (roGFP) have been used to detect hydrogen peroxide (H₂O₂) levels by fusing exogenous thiol peroxidases, such as Orp1 and Tsa2. However, the effectiveness of these thiol peroxidases compared to endogenous ones remains unexplored. Here, we develop a H₂O₂ probe by fusing roGFP2 to an endogenous H₂O₂ sensor, type II peroxiredoxin (PRXIIB), which displayed enhanced responsiveness and conversion kinetics compared to roGFP2-Orp1 in vitro and superior sensitivity to H₂O₂ in vivo. The roGFP2-PRXIIB probe allowed robust visualization of H₂O₂ production in abiotic and biotic stresses, and growing pollen tubes. We further targeted roGFP2-PRXIIB to cytosol, nuclei, mitochondria and chloroplasts to monitor H₂O₂ accumulation in real time in different subcellular compartments during immune activation, and the analyses revealed different temporal patterns of H₂O₂ accumulation during pattern- and effector-triggered immune responses in different compartments. Taken together, the work provides an ultra-sensitive probe for H₂O₂ dynamics in diverse plant biological processes.

Hidden hunger, caused by chronic micronutrient deficiencies, affects billions of people worldwide and remains a critical public health issue despite progress in food production. Biofortification offers a promising solution by enhancing nutrient levels within plant tissues through traditional breeding or advanced biotechnologies. Recent advancements in plant synthetic biology have significantly improved biofortification strategies, enabling precise and targeted nutrient enrichment. This mini-review outlines five core strategies in synthetic biology-based biofortification: overexpression of endogenous biosynthetic genes, introduction of heterologous biosynthetic pathways, expression of nutrient-specific transporters, optimization of transcriptional regulation, and protein (directed) evolution. Vitamin B₁ biofortification serves as a primary illustrative example due to its historical importance and ongoing relevance. Recent breakthroughs, particularly from Chinese research teams, are also highlighted. Together, these strategies offer transformative potential for addressing global nutritional challenges through precise, sustainable and innovative plant-based approaches.

Transgenic and gene-editing technologies are essential for gene functional analysis and crop improvement. However, the pleiotropic effects and unknown mechanisms of morphogenic genes have hindered their broader application. In this study, we employed the one-step de novo shoot organogenesis (DNSO) method, and demonstrated that overexpression of the morphogenic gene Arabidopsis thanalia GROWTH-REGULATING FACTOR 5 (AtGRF5) significantly enhanced genetic transformation efficiency in cucurbit crops by promoting callus proliferation and increasing dense cells during regeneration. High-resolution time-series transcriptomics and single-cell RNA sequencing revealed that AtGRF5 overexpression induced auxin-related genes and expanded stem cell populations during cucumber DNSO. Using DNA-affinity purification sequencing (DAP-seq) in combination with spatiotemporal differential gene expression analysis, we identified CsIAA19 as a key downstream target of AtGRF5, with its modulation playing a pivotal role in regeneration. Rescuing CsIAA19 in AtGRF5-overexpressing explant reversed the enhanced callus proliferation and regeneration. To address growth defects caused by AtGRF5 overexpression, we developed an abscisic acid-inducible AtGRF5 expression system, significantly improving transformation and gene-editing efficiency across diverse genotypes while minimizing pleiotropic effects. In summary, this research provides mechanistic insights into AtGRF5-mediated transformation and offers a practical solution to overcome challenges in cucurbit crop genetic modification.

Seedlessness is a most valuable trait in fruit crops for fresh consumption and processing. The mutations in essential meiosis genes are known to confer sterility and seed abortion in plants. However, defects in meiosis have rarely been reported in fruit crops. Here, we found meiosis defects caused sterility in a seedless citrus bud sport cultivar, with massive unpaired univalents during diakinesis, indicating a disruption in crossover formation. A non-functional CrMER3A^{-103 bp} allele with a 103-bp deletion in the gene body, together with the other non-functional CrMER3a allele with a T deletion in exon, were identified in the seedless cultivar. The CrMER3 protein was undetectable at meiotic prophase I in the seedless cultivar, and knock out of CrMER3 resulted in sterility in precocious Mini-citrus. Therefore, the natural variation in CrMER3 is responsible for sterility and seedlessness in this bud sport cultivar. The CrMER3a allele originated from the primitive wild mandarin and was passed to cultivated mandarins. A Kompetitive Allele-Specific PCR (KASP) marker was developed to identify citrus germplasm with CrMER3a allele and to screen potential sterile and seedless hybrids in citrus cross breeding. Uncovering the natural mutations responsible for meiosis defects in citrus enhances our understanding of mechanisms controlling seedlessness in fruit crops and facilitates breeding of seedless varieties.

Rice production is severely impacted by pathogens such as Magnaporthe oryzae and the rice stripe virus (RSV). Ineffectiveness in controlling viruses and the excessive use of fungicides have proven traditional chemical pesticides increasingly inadequate. RNA interference (RNAi) represents a cutting-edge approach for combating crop diseases, especially in rice. This study addresses the critical gap in scalable, effective RNAi-based rice disease management by exploring the potential of spray-applied small RNA (sRNA) and double-stranded RNA (dsRNA) molecules. We utilized dsRNAs produced by in vitro transcription and bacterial expression systems and employed layered double hydroxides (LDH) to enhance RNA stability, absorption, and efficacy. Our research demonstrated that modified sRNAs could effectively penetrate M. oryzae cell membranes and inhibit conidial germination and appressorium formation, while LDH-conjugated dsRNAs provided prolonged and enhanced protection against both rice blast and rice stripe diseases. Most importantly, dsRNA treatments resulted in improved agronomic traits or increased crop yields by protecting against blast and stripe diseases. This study also validated the compatibility of these RNA molecules with industrial production methods, highlighting their potential as a scalable and eco-friendly option for managing crop diseases at the gene level. This work not only offers a new direction for rice disease control but also provides a foundation for the broader application of RNAi technology in agricultural pest management.

RNA interference (RNAi) is increasingly used for plant protection against pathogens and pests. However, the traditional delivery method causes plant tissue damage, is affected by environmental factors, and faces difficulties in penetrating the barriers of cell walls and the limitations of plant species, ultimately leading to low delivery efficiency. With advances in nanotechnology, nanomaterials (NMs) have been identified as effective carriers for nucleic acid delivery because of their ability to operate independently of external mechanical forces, prevent degradation by bioenzymes, exhibit good biocompatibility, and offer high loading capacity. This review summarizes the application of NM-mediated RNAi against plant pathogens and pests, focusing on how different NMs break through the cell barriers of plants, pathogens, and pests according to their size, morphology, and charge characteristics. Furthermore, we discuss the advantages and improvement strategies of NMs as nucleic acid delivery carriers, alongside assessing their potential application for the management of plant pathogens and pests.

Carbon assimilation is a crucial part of the photosynthetic process, wherein inorganic carbon, typically in the form of CO₂, is converted into organic compounds by living organisms, including plants, algae, and a subset of bacteria. Although several carbon fixation pathways have been elucidated, the Calvin-Benson-Bassham (CBB) cycle remains fundamental to carbon metabolism, playing a pivotal role in the biosynthesis of starch and sucrose in plants, algae, and cyanobacteria. However, Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the key carboxylase enzyme of the CBB cycle, exhibits low kinetic efficiency, low substrate specificity, and high temperature sensitivity, all of which have the potential to limit flux through this pathway. Consequently, RuBisCO needs to be present at very high concentrations, which is one of the factors contributing to its status as the most prevalent protein on Earth. Numerous attempts have been made to optimize the catalytic efficiency of RuBisCO and thereby promote plant growth. Furthermore, the limitations of this process highlight the potential benefits of engineering or discovering more efficient carbon fixation mechanisms, either by improving RuBisCO itself or by introducing alternative pathways. Here, we review advances in artificial carbon assimilation engineering, including the integration of synthetic biology, genetic engineering, metabolic pathway optimization, and artificial intelligence in order to create plants capable of performing more efficient photosynthesis. We additionally provide a perspective of current challenges and potential solutions alongside a personal opinion of the most promising future directions of this emerging field.

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.

The past decade has witnessed rapid developments in gene discovery, biological big data (BBD), artificial intelligence (AI)-aided technologies, and molecular breeding. These advancements are expected to accelerate crop breeding under the pressure of increasing demands for food. Here, we first summarize current breeding methods and discuss the need for new ways to support breeding efforts. Then, we review how to combine BBD and AI technologies for genetic dissection, exploring functional genes, predicting regulatory elements and functional domains, and phenotypic prediction. Finally, we propose the concept of intelligent precision design breeding (IPDB) driven by AI technology and offer ideas about how to implement IPDB. We hope that IPDB will enhance the predictability, efficiency, and cost of crop breeding compared with current technologies. As an example of IPDB, we explore the possibilities offered by CropGPT, which combines biological techniques, bioinformatics, and breeding art from breeders, and presents an open, shareable, and cooperative breeding system. IPDB provides integrated services and communication platforms for biologists, bioinformatics experts, germplasm resource specialists, breeders, dealers, and farmers, and should be well suited for future breeding.

In plants, proteins are fundamental to virtually all biological processes, such as photosynthesis, signal transduction, metabolic regulation, and stress responses. Studying protein distribution, function, modifications, and interactions at the cellular and tissue levels is critical for unraveling the complexities of these biological pathways. Protein abundance and localization are highly dynamic and vary widely across the proteome, presenting a challenge for global protein quantification and analysis. Mass spectrometry-based proteomics approaches have proven to be powerful tools for addressing this complex issue. In this review, we summarize recent advancements in proteomics research and their applications in plant biology, with an emphasis on the current state and challenges of studying post-translational modifications, single-cell proteomics, and protein–protein interactions. Additionally, we discuss future prospects for plant proteomics, highlighting potential opportunities that proteomics technologies offer in advancing plant biology research.

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.

The modern cultivated tomato (Solanum lycopersicum) was domesticated from Solanum pimpinellifolium native to the Andes Mountains of South America through a “two-step domestication” process. It was introduced to Europe in the 16th century and later widely cultivated worldwide. Since the late 19th century, breeders, guided by modern genetics, breeding science, and statistical theory, have improved tomatoes into an important fruit and vegetable crop that serves both fresh consumption and processing needs, satisfying diverse consumer demands. Over the past three decades, advancements in modern crop molecular breeding technologies, represented by molecular marker technology, genome sequencing, and genome editing, have significantly transformed tomato breeding paradigms. This article reviews the research progress in the field of tomato molecular breeding, encompassing genome sequencing of germplasm resources, the identification of functional genes for agronomic traits, and the development of key molecular breeding technologies. Based on these advancements, we also discuss the major challenges and perspectives in this field.

The utilization of metabolomics approaches to explore the metabolic mechanisms underlying plant fitness and adaptation to dynamic environments is growing, highlighting the need for an efficient and user-friendly toolkit tailored for analyzing the extensive datasets generated by metabolomics studies. Current protocols for metabolome data analysis often struggle with handling large-scale datasets or require programming skills. To address this, we present MetMiner (https://github.com/ShawnWx2019/MetMiner), a user-friendly, full-functionality pipeline specifically designed for plant metabolomics data analysis. Built on R shiny, MetMiner can be deployed on servers to utilize additional computational resources for processing large-scale datasets. MetMiner ensures transparency, traceability, and reproducibility throughout the analytical process. Its intuitive interface provides robust data interaction and graphical capabilities, enabling users without prior programming skills to engage deeply in data analysis. Additionally, we constructed and integrated a plant-specific mass spectrometry database into the MetMiner pipeline to optimize metabolite annotation. We have also developed MDAtoolkits, which include a complete set of tools for statistical analysis, metabolite classification, and enrichment analysis, to facilitate the mining of biological meaning from the datasets. Moreover, we propose an iterative weighted gene co-expression network analysis strategy for efficient biomarker metabolite screening in large-scale metabolomics data mining. In two case studies, we validated MetMiner's efficiency in data mining and robustness in metabolite annotation. Together, the MetMiner pipeline represents a promising solution for plant metabolomics analysis, providing a valuable tool for the scientific community to use with ease.

Halorhodospira (Hlr.) halochloris is a triply extremophilic phototrophic purple sulfur bacterium, as it is thermophilic, alkaliphilic, and extremely halophilic. The light-harvesting-reaction center (LH1-RC) core complex of this bacterium displays an LH1-Q_y transition at 1,016 nm, which is the lowest-energy wavelength absorption among all known phototrophs. Here we report the cryo-EM structure of the LH1-RC at 2.42 Å resolution. The LH1 complex forms a tricyclic ring structure composed of 16 αβγ-polypeptides and one αβ-heterodimer around the RC. From the cryo-EM density map, two previously unrecognized integral membrane proteins, referred to as protein G and protein Q, were identified. Both of these proteins are single transmembrane-spanning helices located between the LH1 ring and the RC L- subunit and are absent from the LH1-RC complexes of all other purple bacteria of which the structures have been determined so far. Besides bacteriochlorophyll b molecules (B1020) located on the periplasmic side of the Hlr. halochloris membrane, there are also two arrays of bacteriochlorophyll b molecules (B800 and B820) located on the cytoplasmic side. Only a single copy of a carotenoid (lycopene) was resolved in the Hlr. halochloris LH1-α3β3 and this was positioned within the complex. The potential quinone channel should be the space between the LH1-α3β3 that accommodates the single lycopene but does not contain a γ-polypeptide, B800 and B820. Our results provide a structural explanation for the unusual Q_y red shift and carotenoid absorption in the Hlr. halochloris spectrum and reveal new insights into photosynthetic mechanisms employed by a species that thrives under the harshest conditions of any phototrophic microorganism known.

By 2050, the global population is projected to reach 9 billion, underscoring the imperative for innovative solutions to increase grain yield and enhance food security. Nanotechnology has emerged as a powerful tool, providing unique solutions to this challenge. Nanoparticles (NPs) can improve plant growth and nutrition under normal conditions through their high surface-to-volume ratio and unique physical and chemical properties. Moreover, they can be used to monitor crop health status and augment plant resilience against abiotic stresses (such as salinity, drought, heavy metals, and extreme temperatures) that endanger global agriculture. Application of NPs can enhance stress tolerance mechanisms in plants, minimizing potential yield losses and underscoring the potential of NPs to raise crop yield and quality. This review highlights the need for a comprehensive exploration of the environmental implications and safety of nanomaterials and provides valuable guidelines for researchers, policymakers, and agricultural practitioners. With thoughtful stewardship, nanotechnology holds immense promise in shaping environmentally sustainable agriculture amid escalating environmental challenges.