Selenium (Se) is a vital micronutrient for humans, with important functions for health and anti‐cancer properties. Organic Se shows higher antioxidant activity and much lower toxicity compared to inorganic Se, making it safer for use. Selenomethionine (SeMet) is one of the primary forms of organic Se. OsPT4, the high‐affinity phosphate (Pi) transporter (PHT) of rice, has been investigated for its role in the transport of the different forms of Se, and its effects on the accumulation of SeMet in this study. The OsPT4 mutant and overexpression lines were used as research materials. Phenotypic analyses revealed that OsPT4 confers improved Se tolerance in shoots upon selenite exposure. Heterologous expression assays in Xenopus laevis oocytes and yeast systems and translocation assays in different transgenic lines of rice confirmed OsPT4 ‐mediated selenite and SeMet transport activity, establishing its responsibility for root‐to‐shoot Se translocation. Transcriptomic profiling, amino acid quantification and qRT‐PCR analyses further indicated that OsPT4 up‐regulates methionine (Met) biosynthesis, the direct precursor of SeMet. Notably, OsPT4 significantly increased SeMet accumulation and promoted the formation of Se‐rich micron‐sized spherical particles in seeds under Se supplementation. These findings provide mechanistic insights into OsPT4 ‐mediated SeMet trafficking and metabolism, advancing strategies for developing Se‐biofortified rice cultivars with enhanced nutritional and therapeutic value.
{"title":"OsPT4 Facilitates Selenomethionine Transport and Biosynthesis to Enhance Seed Accumulation in Rice: Molecular Mechanisms and Biotechnological Potential","authors":"Yang Yang, Lijuan Sun, Jia Wei, Fang Zhang, Shiyan Yang, Jiarui Zhang, Qin Qin, Jun Wang, Guohua Xu, Shubin Sun, Yafei Sun, Yong Xue","doi":"10.1111/pbi.70465","DOIUrl":"https://doi.org/10.1111/pbi.70465","url":null,"abstract":"Selenium (Se) is a vital micronutrient for humans, with important functions for health and anti‐cancer properties. Organic Se shows higher antioxidant activity and much lower toxicity compared to inorganic Se, making it safer for use. Selenomethionine (SeMet) is one of the primary forms of organic Se. OsPT4, the high‐affinity phosphate (Pi) transporter (PHT) of rice, has been investigated for its role in the transport of the different forms of Se, and its effects on the accumulation of SeMet in this study. The <jats:italic>OsPT4</jats:italic> mutant and overexpression lines were used as research materials. Phenotypic analyses revealed that <jats:italic>OsPT4</jats:italic> confers improved Se tolerance in shoots upon selenite exposure. Heterologous expression assays in <jats:styled-content style=\"fixed-case\"> <jats:italic>Xenopus laevis</jats:italic> </jats:styled-content> oocytes and yeast systems and translocation assays in different transgenic lines of rice confirmed <jats:italic>OsPT4</jats:italic> ‐mediated selenite and SeMet transport activity, establishing its responsibility for root‐to‐shoot Se translocation. Transcriptomic profiling, amino acid quantification and qRT‐PCR analyses further indicated that OsPT4 up‐regulates methionine (Met) biosynthesis, the direct precursor of SeMet. Notably, <jats:italic>OsPT4</jats:italic> significantly increased SeMet accumulation and promoted the formation of Se‐rich micron‐sized spherical particles in seeds under Se supplementation. These findings provide mechanistic insights into <jats:italic>OsPT4</jats:italic> ‐mediated SeMet trafficking and metabolism, advancing strategies for developing Se‐biofortified rice cultivars with enhanced nutritional and therapeutic value.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"126 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145515605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
I summarise and discuss the effects of environmental factors and culture conditions on the productivity of recombinant proteins, including biopharmaceuticals, in Nicotiana benthamiana via transient gene expression platforms. A primary focus is on controlled‐environment plant cultivation systems such as plant factories with artificial lighting. Before gene transfer, sufficient leaf biomass accumulation is suggested to contribute positively to subsequent target protein expression per unit of leaf biomass. Increasing the plant density and extending the photoperiod may be simple methods for increasing biomass productivity per unit of cultivation area; however, careful evaluation is needed to determine the effectiveness of these methods. Strictly regulated plant growth conditions should be crucial not only after but also before gene transfer to reduce batch‐to‐batch variation in target protein productivity. After gene transfer, target protein accumulation in leaves is highly sensitive to temperature. In this context, not only the air temperature but also the leaf temperature should be considered, given that the two temperatures can differ as a result of the leaf energy balance. Agroinfiltrated leaves often exhibit necrosis, which has been suggested to be triggered by endoplasmic reticulum stress, followed by oxidative stress. Lowering the air temperature or applying ascorbic acid can reduce the severity of necrosis. Light may only be necessary for specific timings after agroinfiltration, although more work is needed to draw general conclusions. A detached leaf‐based system may be a promising choice because the handling of plant materials is easier than a whole plant‐based system. Finally, current conclusions and future perspectives are provided.
{"title":"The Impacts of Environmental Factors and Culture Conditions on the Transient Expression of Recombinant Protein in Nicotiana benthamiana Under Controlled Environments","authors":"Ryo Matsuda","doi":"10.1111/pbi.70453","DOIUrl":"https://doi.org/10.1111/pbi.70453","url":null,"abstract":"I summarise and discuss the effects of environmental factors and culture conditions on the productivity of recombinant proteins, including biopharmaceuticals, in <jats:italic>Nicotiana benthamiana</jats:italic> via transient gene expression platforms. A primary focus is on controlled‐environment plant cultivation systems such as plant factories with artificial lighting. Before gene transfer, sufficient leaf biomass accumulation is suggested to contribute positively to subsequent target protein expression per unit of leaf biomass. Increasing the plant density and extending the photoperiod may be simple methods for increasing biomass productivity per unit of cultivation area; however, careful evaluation is needed to determine the effectiveness of these methods. Strictly regulated plant growth conditions should be crucial not only after but also before gene transfer to reduce batch‐to‐batch variation in target protein productivity. After gene transfer, target protein accumulation in leaves is highly sensitive to temperature. In this context, not only the air temperature but also the leaf temperature should be considered, given that the two temperatures can differ as a result of the leaf energy balance. Agroinfiltrated leaves often exhibit necrosis, which has been suggested to be triggered by endoplasmic reticulum stress, followed by oxidative stress. Lowering the air temperature or applying ascorbic acid can reduce the severity of necrosis. Light may only be necessary for specific timings after agroinfiltration, although more work is needed to draw general conclusions. A detached leaf‐based system may be a promising choice because the handling of plant materials is easier than a whole plant‐based system. Finally, current conclusions and future perspectives are provided.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"60 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145515606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiao‐Wen Wang, Li Ling, Ke‐Zheng Zhao, Jiangqi Wen, Zhaolin Ji, Xiao‐Ren Chen, Feng Zhu
Plant viruses are so harmful to crops. It is an urgent need to develop modern, environment‐friendly, and sustainable plant viral epidemic‐management strategies that are safe for both human health and the environment. The field of nanotechnology is gaining increased interest in plant science. Magnesium oxide nanoparticles (MgONPs) have typical physical and chemical characteristics of nanomaterials. Hence, in this study, we systematically investigated the molecular mechanism of MgONPs triggering the plant immunity against viral pathogens. Foliar treatments allow MgONPs to enter Nicotiana benthamiana leaves through stomata and distribute within the intracellular space around chloroplasts through penetrating cell walls. MgONPs elevate plant growth and trigger dose‐dependent plant immunity against viral pathogens. Application of MgONPs triggers glutamate‐like receptors‐dependent Ca 2+ flux and Ca 2+ sensors. Exogenous application of MgONPs does not trigger resistance in Ca 2+ channel‐blocked plants, and knockout of NbGLR3.3 weakens the resistance induced by MgONPs. MgONPs induce early ROS bursts but reduce oxidative damage and accumulation of ROS after TMV infection at late stages. MgONPs activate Ca 2+ ‐dependent SA‐, JA‐, and ET‐mediated signalling pathways, and the absence of SA‐, JA‐, or ET signals weakens the MgONPs‐triggered resistance. However, MgONPs fail to induce resistance to viral pathogens in plants simultaneously lacking SA, JA, and ET. Safety evaluation showed that MgONPs have desirable biocompatibility and biosafety for plants, as well as satisfactory biosafety for the aquatic environment. Overall, our discoveries point to a new direction for MgONPs as effective, non‐drug‐resistant, non‐toxic, sustainable, residual‐free, and eco‐friendly antiviral agents to simultaneously prevent diverse viral diseases.
{"title":"Join the Green and Sustainable Team: Magnesium Oxide Nanoparticles Boost Broad‐Spectrum Viral Resistance in Solanaceae Plants","authors":"Xiao‐Wen Wang, Li Ling, Ke‐Zheng Zhao, Jiangqi Wen, Zhaolin Ji, Xiao‐Ren Chen, Feng Zhu","doi":"10.1111/pbi.70461","DOIUrl":"https://doi.org/10.1111/pbi.70461","url":null,"abstract":"Plant viruses are so harmful to crops. It is an urgent need to develop modern, environment‐friendly, and sustainable plant viral epidemic‐management strategies that are safe for both human health and the environment. The field of nanotechnology is gaining increased interest in plant science. Magnesium oxide nanoparticles (MgONPs) have typical physical and chemical characteristics of nanomaterials. Hence, in this study, we systematically investigated the molecular mechanism of MgONPs triggering the plant immunity against viral pathogens. Foliar treatments allow MgONPs to enter <jats:italic>Nicotiana benthamiana</jats:italic> leaves through stomata and distribute within the intracellular space around chloroplasts through penetrating cell walls. MgONPs elevate plant growth and trigger dose‐dependent plant immunity against viral pathogens. Application of MgONPs triggers glutamate‐like receptors‐dependent Ca <jats:sup>2+</jats:sup> flux and Ca <jats:sup>2+</jats:sup> sensors. Exogenous application of MgONPs does not trigger resistance in Ca <jats:sup>2+</jats:sup> channel‐blocked plants, and knockout of <jats:italic>NbGLR3.3</jats:italic> weakens the resistance induced by MgONPs. MgONPs induce early ROS bursts but reduce oxidative damage and accumulation of ROS after TMV infection at late stages. MgONPs activate Ca <jats:sup>2+</jats:sup> ‐dependent SA‐, JA‐, and ET‐mediated signalling pathways, and the absence of SA‐, JA‐, or ET signals weakens the MgONPs‐triggered resistance. However, MgONPs fail to induce resistance to viral pathogens in plants simultaneously lacking SA, JA, and ET. Safety evaluation showed that MgONPs have desirable biocompatibility and biosafety for plants, as well as satisfactory biosafety for the aquatic environment. Overall, our discoveries point to a new direction for MgONPs as effective, non‐drug‐resistant, non‐toxic, sustainable, residual‐free, and eco‐friendly antiviral agents to simultaneously prevent diverse viral diseases.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"39 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145509423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Diosmin is often used as a dietary supplement, and it is sometimes used to make phlebotropic drugs for venous diseases. Traditionally, diosmin is obtained by the dehydrogenation of the flavanone glycoside hesperidin through chemical semisynthesis. In this study, we identified and characterised a flavonoid‐O‐methyltransferase (F4′OMT) and two glucosyltransferases (F7GlcTs) and completed the biosynthetic pathway of diosmin and its analogue, linarin and isorhoifolin, in combination with the previously found flavone synthase (FNSII) and rhamnosyltransferase (RhaT). We developed an isocaudarner‐based gene stacking strategy to integrate FNS, OMT, GlcT and RhaT into a multi‐gene vector that was transformed into Solanum lycopersicum cv. ‘MicroTom’ and Nicotiana tabacum . The introduction of the flavone pathway enabled the production of diosmin at 474 ng/g DW in the peel of transgenic tomato and 20.5 ng/g DW in the leaves of tobacco. In addition, linarin with a 299.5 ng/g level and isorhoifolin with a 2.8 ng/g level can be found in the leaves of tobacco. How the endogenous enzymes and metabolic flux affected the production of the specialised compounds was discussed. Our results represent the first attempt in the heterologous biosynthesis of O‐methylated and rhamnosylated flavone di‐glucosides in flavonol‐producing plants.
地奥司明常被用作膳食补充剂，有时也被用于制造静脉疾病的促静脉药物。传统上，薯蓣皂苷是由黄酮苷橙皮苷通过化学半合成脱氢得到的。在这项研究中，我们鉴定并鉴定了一种类黄酮- O -甲基转移酶（F4'OMT）和两种葡萄糖基转移酶（F7GlcTs），并与先前发现的黄酮合成酶（FNSII）和鼠李糖基转移酶（RhaT）结合，完成了二元草明及其类似物亚麻素和异花叶素的生物合成途径。我们开发了一种基于异源性的基因堆叠策略，将FNS、OMT、GlcT和RhaT整合到一个多基因载体中，并将其转化为番茄植株。“MicroTom”和烟草。黄酮途径的引入使转基因番茄果皮中薯蓣皂苷的产量为474 ng/g DW，烟草叶片中薯蓣皂苷的产量为20.5 ng/g DW。此外，在烟草叶片中还可发现亚麻素含量为299.5 ng/g，异油烟素含量为2.8 ng/g。讨论了内源性酶和代谢通量如何影响特异性化合物的产生。我们的研究结果是在黄酮醇产生植物中异种生物合成O -甲基化和鼠李糖化黄酮二糖苷的第一次尝试。
{"title":"Engineering of the Flavone Rutinoside Biosynthetic Pathway Enables Production of Diosmin, a Venoactive Compound in Solanum lycopersicum and Nicotiana tabacum ","authors":"Qianle Zhang, Qingwen Wu, Wenjing Shi, Yinai Deng, Ruoting Zhan, Dongming Ma","doi":"10.1111/pbi.70426","DOIUrl":"https://doi.org/10.1111/pbi.70426","url":null,"abstract":"Diosmin is often used as a dietary supplement, and it is sometimes used to make phlebotropic drugs for venous diseases. Traditionally, diosmin is obtained by the dehydrogenation of the flavanone glycoside hesperidin through chemical semisynthesis. In this study, we identified and characterised a flavonoid‐O‐methyltransferase (F4′OMT) and two glucosyltransferases (F7GlcTs) and completed the biosynthetic pathway of diosmin and its analogue, linarin and isorhoifolin, in combination with the previously found flavone synthase (FNSII) and rhamnosyltransferase (RhaT). We developed an isocaudarner‐based gene stacking strategy to integrate FNS, OMT, GlcT and RhaT into a multi‐gene vector that was transformed into <jats:styled-content style=\"fixed-case\"> <jats:italic>Solanum lycopersicum</jats:italic> </jats:styled-content> cv. ‘MicroTom’ and <jats:styled-content style=\"fixed-case\"> <jats:italic>Nicotiana tabacum</jats:italic> </jats:styled-content> . The introduction of the flavone pathway enabled the production of diosmin at 474 ng/g DW in the peel of transgenic tomato and 20.5 ng/g DW in the leaves of tobacco. In addition, linarin with a 299.5 ng/g level and isorhoifolin with a 2.8 ng/g level can be found in the leaves of tobacco. How the endogenous enzymes and metabolic flux affected the production of the specialised compounds was discussed. Our results represent the first attempt in the heterologous biosynthesis of O‐methylated and rhamnosylated flavone di‐glucosides in flavonol‐producing plants.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"2 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145498388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Philip Alexander Heine, Tetyana Nosenko, Sarah Kistner, Kevin Dennis Oliphant, Manuel Hanke‐Uhe, Afsheen Shahid, Bin Hu, Martin Kucklick, Nina Lehmler, Marlies Becker, Nicole Goerke, Janin Korn, Tanja Linke, Doris Meier, Asta Perl, Saskia Polten, Valeska Priess, Dorina Schäckermann, Maren Schubert, Jörg Schumacher, Jana Barbro Winkler, Susanne Engelmann, Heinz Rennenberg, Jörg‐Peter Schnitzler, Stefan Dübel, Michael Hust, Robert Hänsch, David Kaufholdt
Plant defensins (PDFs) are cysteine‐rich antimicrobial peptides (AMPs) that are important components of plant immunity. They occur constitutively in various plant tissues but are also upregulated upon stress. Therefore, these molecules are of great interest as markers for the diagnosis of early forest stress response in plants at the molecular level. PDFs are small peptides (~5 kDa) with a compact tertiary structure, requiring specific protocols and dedicated antibodies for detection by quantitative ELISA. We developed monoclonal recombinant antibodies using phage display in solution against the correctly folded antigen defensin Fs PDF2 from beech ( Fagus sylvatica ) and analysed the antibody–antigen interaction in silico with AlphaFold 3. In a proof‐of‐principle study, we investigated the Fs PDF2 stress response to abiotic (drought) and biotic (gall midge) stresses. Notably, we established an assay for defensin quantification in crude plant extract, detecting for the first time natively folded proteins in a specific sandwich ELISA. Our antibody generation strategy can be transferred by practitioners to other small antimicrobial peptides (AMP), paving the way to study this group of proteins and their corresponding stress response comprehensively.
{"title":"Phage Display Derived Antibodies Against Antimicrobial Peptide Fs PDF2 Reveal Stress Response in European Beech","authors":"Philip Alexander Heine, Tetyana Nosenko, Sarah Kistner, Kevin Dennis Oliphant, Manuel Hanke‐Uhe, Afsheen Shahid, Bin Hu, Martin Kucklick, Nina Lehmler, Marlies Becker, Nicole Goerke, Janin Korn, Tanja Linke, Doris Meier, Asta Perl, Saskia Polten, Valeska Priess, Dorina Schäckermann, Maren Schubert, Jörg Schumacher, Jana Barbro Winkler, Susanne Engelmann, Heinz Rennenberg, Jörg‐Peter Schnitzler, Stefan Dübel, Michael Hust, Robert Hänsch, David Kaufholdt","doi":"10.1111/pbi.70431","DOIUrl":"https://doi.org/10.1111/pbi.70431","url":null,"abstract":"Plant defensins (PDFs) are cysteine‐rich antimicrobial peptides (AMPs) that are important components of plant immunity. They occur constitutively in various plant tissues but are also upregulated upon stress. Therefore, these molecules are of great interest as markers for the diagnosis of early forest stress response in plants at the molecular level. PDFs are small peptides (~5 kDa) with a compact tertiary structure, requiring specific protocols and dedicated antibodies for detection by quantitative ELISA. We developed monoclonal recombinant antibodies using phage display in solution against the correctly folded antigen defensin <jats:italic>Fs</jats:italic> PDF2 from beech ( <jats:styled-content style=\"fixed-case\"> <jats:italic>Fagus sylvatica</jats:italic> </jats:styled-content> ) and analysed the antibody–antigen interaction in silico with AlphaFold 3. In a proof‐of‐principle study, we investigated the <jats:italic>Fs</jats:italic> PDF2 stress response to abiotic (drought) and biotic (gall midge) stresses. Notably, we established an assay for defensin quantification in crude plant extract, detecting for the first time natively folded proteins in a specific sandwich ELISA. Our antibody generation strategy can be transferred by practitioners to other small antimicrobial peptides (AMP), paving the way to study this group of proteins and their corresponding stress response comprehensively.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"144 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145498383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Oomycete pathogens secrete hundreds of RXLR effectors into plant cells to modulate host immunity by targeting diverse plant proteins. Here, we report that the Peronophythora litchii RXLR effector PlAvh133 acts as a virulence factor and targets the litchi glycolate oxidase (GLO) LcGLO1, a key enzyme in photorespiration, thereby suppressing plant immunity. PlAvh133 localises to the plasma membrane (PM) in planta, and its first α-helix is vital for both its LcGLO1-binding activity and proper PM localisation. LcGLO1 is mainly confined to the peroxisomes, and its overexpression significantly enhanced resistance to downy blight in litchi. Conversely, silencing the Nicotiana benthamiana homologue of LcGLO1 increases plant susceptibility to the oomycete pathogen. Critically, PlAvh133 causes the relocation of LcGLO1 from peroxisomes to the PM and inhibits its enzymatic activity, leading to increased plant susceptibility. PM-localised LcGLO1 cooperates with catalase (CAT) LcCATB to suppress reactive oxygen species (ROS) burst. Meanwhile, PM-localised LcGLO1 destabilises respiratory burst oxidase homologue (RBOH) LcRBOHD by interacting with calcium-dependent protein kinase (CPK) LcCPK5, further reducing ROS production. Taken together, our findings unveil an unprecedented virulence mechanism by which a pathogen effector relocalises and inhibits host GLO1 activity, thereby simultaneously diminishing ROS production from both the peroxisomes and PM-localised RBOHD.
{"title":"Effector-Mediated Spatial Reprogramming of Glycolate Oxidase Subverts Peroxisomal and Membrane-Associated ROS Defences.","authors":"Junjian Situ,Zijing Zhang,Yi Shao,Jiaying Feng,Feiteng Zhong,Muran Xiong,Wen Li,Peng Li,Xiaofan Zhou,Guibing Hu,Jietang Zhao,Minhui Li,Pinggen Xi,Xinxiang Peng,Zide Jiang,Guanghui Kong","doi":"10.1111/pbi.70457","DOIUrl":"https://doi.org/10.1111/pbi.70457","url":null,"abstract":"Oomycete pathogens secrete hundreds of RXLR effectors into plant cells to modulate host immunity by targeting diverse plant proteins. Here, we report that the Peronophythora litchii RXLR effector PlAvh133 acts as a virulence factor and targets the litchi glycolate oxidase (GLO) LcGLO1, a key enzyme in photorespiration, thereby suppressing plant immunity. PlAvh133 localises to the plasma membrane (PM) in planta, and its first α-helix is vital for both its LcGLO1-binding activity and proper PM localisation. LcGLO1 is mainly confined to the peroxisomes, and its overexpression significantly enhanced resistance to downy blight in litchi. Conversely, silencing the Nicotiana benthamiana homologue of LcGLO1 increases plant susceptibility to the oomycete pathogen. Critically, PlAvh133 causes the relocation of LcGLO1 from peroxisomes to the PM and inhibits its enzymatic activity, leading to increased plant susceptibility. PM-localised LcGLO1 cooperates with catalase (CAT) LcCATB to suppress reactive oxygen species (ROS) burst. Meanwhile, PM-localised LcGLO1 destabilises respiratory burst oxidase homologue (RBOH) LcRBOHD by interacting with calcium-dependent protein kinase (CPK) LcCPK5, further reducing ROS production. Taken together, our findings unveil an unprecedented virulence mechanism by which a pathogen effector relocalises and inhibits host GLO1 activity, thereby simultaneously diminishing ROS production from both the peroxisomes and PM-localised RBOHD.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145491580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yonglu Wei, Zengyu Lin, Qi Xie, Jie Gao, Jianpeng Jin, Jie Li, Chuqiao Lu, Guangying Ye, Wenkang Li, Chuanfeng Huang, Dengqi Yang, Qi Liu, Genfa Zhu, Fengxi Yang
The Orchidaceae family, with its unparalleled species diversity among angiosperms, is integral to ornamental, medicinal, cultural, and ecological value. Multi‐omics techniques have proven invaluable for the identification of candidate genes and the advancement of functional genomics research. Nevertheless, the application of these technologies in Orchidaceae remains severely limited due to the lack of effective platforms that can integrate and analyze multi‐omics data, especially in understanding the mechanisms underlying key traits such as distinctive floral morphology. In this study, we present OrchidMD, the Orchid Multi‐omics Database ( www.orchidcomics.com ), a resource platform that integrates data from five omics layers: genomics, transcriptomics, proteomics, metabolomics, and phenomics, encompassing a total of 213 species. OrchidMD is equipped with 18 specialized statistical and analytical tools, and features a user‐friendly interface that facilitates efficient gene mining, multi‐omics data exploration, and integrative interactive analysis. A case study on the comprehensive identification of the pan‐ARF gene family across Orchidaceae species demonstrates the effectiveness and convenience of OrchidMD. Furthermore, experimental validation further shows that transgenic overexpression of CsiARF04 promotes the differentiation and budding of orchid rhizomes. In addition, another case study using gene editing in orchids, CRISPR Design was employed to predict the CsiPDS target site in Cymbidium sinense . Effective editing was subsequently achieved via Agrobacterium ‐mediated delivery of the CRISPR/Cas9 vector into leaves. These results underscore OrchidMD‘s formidable capacity to discern candidate genes associated with salient traits and elucidate their regulatory mechanisms. Thus, OrchidMD serves as a pivotal platform advancing multi‐dimensional biological research and functional genomics in orchids.
{"title":"OrchidMD: An Integrated and User‐Interactive Orchid Multi‐Omics Database for Mining Genes and Biological Research","authors":"Yonglu Wei, Zengyu Lin, Qi Xie, Jie Gao, Jianpeng Jin, Jie Li, Chuqiao Lu, Guangying Ye, Wenkang Li, Chuanfeng Huang, Dengqi Yang, Qi Liu, Genfa Zhu, Fengxi Yang","doi":"10.1111/pbi.70445","DOIUrl":"https://doi.org/10.1111/pbi.70445","url":null,"abstract":"The Orchidaceae family, with its unparalleled species diversity among angiosperms, is integral to ornamental, medicinal, cultural, and ecological value. Multi‐omics techniques have proven invaluable for the identification of candidate genes and the advancement of functional genomics research. Nevertheless, the application of these technologies in Orchidaceae remains severely limited due to the lack of effective platforms that can integrate and analyze multi‐omics data, especially in understanding the mechanisms underlying key traits such as distinctive floral morphology. In this study, we present OrchidMD, the Orchid Multi‐omics Database ( <jats:ext-link xmlns:xlink=\"http://www.w3.org/1999/xlink\" xlink:href=\"http://www.orchidcomics.com\">www.orchidcomics.com</jats:ext-link> ), a resource platform that integrates data from five omics layers: genomics, transcriptomics, proteomics, metabolomics, and phenomics, encompassing a total of 213 species. OrchidMD is equipped with 18 specialized statistical and analytical tools, and features a user‐friendly interface that facilitates efficient gene mining, multi‐omics data exploration, and integrative interactive analysis. A case study on the comprehensive identification of the pan‐ARF gene family across Orchidaceae species demonstrates the effectiveness and convenience of OrchidMD. Furthermore, experimental validation further shows that transgenic overexpression of <jats:italic>CsiARF04</jats:italic> promotes the differentiation and budding of orchid rhizomes. In addition, another case study using gene editing in orchids, CRISPR Design was employed to predict the <jats:italic>CsiPDS</jats:italic> target site in <jats:italic>Cymbidium sinense</jats:italic> . Effective editing was subsequently achieved via <jats:italic>Agrobacterium</jats:italic> ‐mediated delivery of the CRISPR/Cas9 vector into leaves. These results underscore OrchidMD‘s formidable capacity to discern candidate genes associated with salient traits and elucidate their regulatory mechanisms. Thus, OrchidMD serves as a pivotal platform advancing multi‐dimensional biological research and functional genomics in orchids.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"377 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145484761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
High‐temperature stress severely threatens rice ( Oryza sativa L.) growth and productivity, particularly during the seedling stage, making heat tolerance an essential breeding target. In this study, we identified the heat shock transcription factor OsHsfc1a as a positive regulator of thermotolerance in rice seedlings. OsHsfc1a expression was strongly induced by heat stress, and overexpression lines exhibited enhanced heat tolerance, whereas knockout mutants were hypersensitive. Integrative transcriptome and DAP‐seq analyses revealed that OsHsfc1a directly binds to and represses the transcription of OsMFT1 , a gene previously implicated in stress responses. Molecular and genetic assays confirmed that OsHsfc1a‐mediated suppression of OsMFT1 helps maintain the chloroplast structural integrity, reduces reactive oxygen species accumulation, and alleviates programmed cell death under heat stress. In contrast, OsMFT1 overexpression impaired chloroplast organization and decreased seedling survival, whereas its knockout enhanced heat tolerance. Furthermore, comparative transcriptome analysis revealed that the OsHsfc1a‐ OsMFT1 regulatory module maintains the chloroplast structural integrity during heat stress by modulating the expression of multiple chloroplast structure‐related genes. Analysis of natural haplotypes revealed that OsHsfc1a allelic variation is associated with rice subspecies differentiation, providing useful genetic resources for breeding heat‐resilient varieties. In summary, we demonstrate that OsHsfc1a enhances heat tolerance by repressing OsMFT1 to preserve chloroplast stability, providing both mechanistic insights and practical genetic resources for rice improvement under global warming.
{"title":"Heat Shock Transcription Factor OsHsfc1a Enhances Rice Seedling Thermotolerance by Regulating OsMFT1 and Preserving Chloroplast Structure Under Heat Stress","authors":"Jingqin Lu, Junyu Chen, Hong Chen, Ziqiang Fan, Lin Lin, Chuanhe Liu, Xiaoxian Wu, Jilei Huang, Zhenlan Liu, Jing Li, Chuxiong Zhuang, Shaoyan Zheng","doi":"10.1111/pbi.70458","DOIUrl":"https://doi.org/10.1111/pbi.70458","url":null,"abstract":"High‐temperature stress severely threatens rice ( <jats:styled-content style=\"fixed-case\"> <jats:italic>Oryza sativa</jats:italic> </jats:styled-content> L.) growth and productivity, particularly during the seedling stage, making heat tolerance an essential breeding target. In this study, we identified the heat shock transcription factor OsHsfc1a as a positive regulator of thermotolerance in rice seedlings. <jats:italic>OsHsfc1a</jats:italic> expression was strongly induced by heat stress, and overexpression lines exhibited enhanced heat tolerance, whereas knockout mutants were hypersensitive. Integrative transcriptome and DAP‐seq analyses revealed that OsHsfc1a directly binds to and represses the transcription of <jats:italic>OsMFT1</jats:italic> , a gene previously implicated in stress responses. Molecular and genetic assays confirmed that OsHsfc1a‐mediated suppression of <jats:italic>OsMFT1</jats:italic> helps maintain the chloroplast structural integrity, reduces reactive oxygen species accumulation, and alleviates programmed cell death under heat stress. In contrast, <jats:italic>OsMFT1</jats:italic> overexpression impaired chloroplast organization and decreased seedling survival, whereas its knockout enhanced heat tolerance. Furthermore, comparative transcriptome analysis revealed that the OsHsfc1a‐ <jats:italic>OsMFT1</jats:italic> regulatory module maintains the chloroplast structural integrity during heat stress by modulating the expression of multiple chloroplast structure‐related genes. Analysis of natural haplotypes revealed that <jats:italic>OsHsfc1a</jats:italic> allelic variation is associated with rice subspecies differentiation, providing useful genetic resources for breeding heat‐resilient varieties. In summary, we demonstrate that OsHsfc1a enhances heat tolerance by repressing <jats:italic>OsMFT1</jats:italic> to preserve chloroplast stability, providing both mechanistic insights and practical genetic resources for rice improvement under global warming.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"203 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145484692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Amélie A Kelly,Martin Fulda,Merle Aden,Ilka N Abreu,Kirstin Feussner,Ivo Feussner
Sinapine (O-sinapoyl choline) is the major phenolic metabolite typically found in the oil-rich seeds of Brassicaceae such as Camelina sativa and Brassica napus. It imparts a bitter taste to the seeds as a defence mechanism against herbivores, but it also renders them less palatable to livestock. To improve Camelina flour for human consumption or as animal feed, we reduced sinapine content through CRISPR/Cas9-based genome editing of REF1 (REDUCED EPIDERMAL FLUORESCENCE1), which encodes the NADP+-dependent coniferaldehyde/sinapaldehyde dehydrogenase (CALDH/SALDH), a key enzyme for sinapine biosynthesis in Arabidopsis thaliana and B. napus. Inactivation of all three homoeologues found in C. sativa lowered the sinapine content in seeds by an overall 56% in two cultivars indicating the presence of a REF1-independent pathway for sinapine biosynthesis. Most importantly however, crucial agronomic seed traits such as total lipid or protein content of the seeds, seed weight or germination were not affected. Hence, the ref1 mutant lines produced here provide a valuable trait, that can be combined with other traits through gene stacking to obtain crops with significantly improved product quality. Furthermore, metabolite fingerprinting by ultra-performance liquid chromatography-electrospray ionisation-quadrupole time-of-flight mass spectrometry of ref1 mutant lines revealed a contrasting phenylpropanoid profile in seeds and leaves, indicating that REF1 oxidises sinapaldehyde to sinapate in seeds and coniferyl aldehyde to ferulate in leaves. In contrast to Arabidopsis however, Camelina accumulates no comparable levels of sinapoyl malate, but substantial amounts of chlorogenic acid, of which an additional chlorogenic acid isomer distinguishes the two different Camelina cultivars as a metabolite marker.
{"title":"Reducing the Sinapine Levels of Camelina sativa Seeds Through Targeted Genome Editing of REF1.","authors":"Amélie A Kelly,Martin Fulda,Merle Aden,Ilka N Abreu,Kirstin Feussner,Ivo Feussner","doi":"10.1111/pbi.70450","DOIUrl":"https://doi.org/10.1111/pbi.70450","url":null,"abstract":"Sinapine (O-sinapoyl choline) is the major phenolic metabolite typically found in the oil-rich seeds of Brassicaceae such as Camelina sativa and Brassica napus. It imparts a bitter taste to the seeds as a defence mechanism against herbivores, but it also renders them less palatable to livestock. To improve Camelina flour for human consumption or as animal feed, we reduced sinapine content through CRISPR/Cas9-based genome editing of REF1 (REDUCED EPIDERMAL FLUORESCENCE1), which encodes the NADP+-dependent coniferaldehyde/sinapaldehyde dehydrogenase (CALDH/SALDH), a key enzyme for sinapine biosynthesis in Arabidopsis thaliana and B. napus. Inactivation of all three homoeologues found in C. sativa lowered the sinapine content in seeds by an overall 56% in two cultivars indicating the presence of a REF1-independent pathway for sinapine biosynthesis. Most importantly however, crucial agronomic seed traits such as total lipid or protein content of the seeds, seed weight or germination were not affected. Hence, the ref1 mutant lines produced here provide a valuable trait, that can be combined with other traits through gene stacking to obtain crops with significantly improved product quality. Furthermore, metabolite fingerprinting by ultra-performance liquid chromatography-electrospray ionisation-quadrupole time-of-flight mass spectrometry of ref1 mutant lines revealed a contrasting phenylpropanoid profile in seeds and leaves, indicating that REF1 oxidises sinapaldehyde to sinapate in seeds and coniferyl aldehyde to ferulate in leaves. In contrast to Arabidopsis however, Camelina accumulates no comparable levels of sinapoyl malate, but substantial amounts of chlorogenic acid, of which an additional chlorogenic acid isomer distinguishes the two different Camelina cultivars as a metabolite marker.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"99 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145477620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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