Seed oil content is a key trait in soybean, yet its genetic basis remains largely unresolved. Here, we identify GmABHD6, an α/β-hydrolase domain-containing gene, as the causal gene underlying a major quantitative trait locus (QTL) for seed oil content on chromosome 16. Through fine-mapping in a chromosome segment substitution population derived from Glycine max (SN14) and wild Glycine soja (ZYD00006), we fine-mapped the QTL to a 150-kb interval. Functional analyses reveal that overexpression of GmABHD6 significantly reduces seed oil content, whereas gene editing to truncate GmABHD6 led to increased oil accumulation. These modifications also alter seed protein levels, seed size and 100-seed weight, indicating pleiotropic effects on seed composition and yield traits. Integrated transcriptomic and metabolomic profiling of transgenic lines highlighted widespread changes in lipid metabolism and energy pathways associated with GmABHD6 activity. Further functional characterisation indicates that the ERF transcription factor GmERF physically binds to the GmABHD6 promoter region to regulate its expression. Population genetic analyses show strong signatures of selection at the GmABHD6 locus, suggesting that favourable alleles are enriched during soybean domestication and improvement. Our findings establish GmABHD6 as a domestication-related gene that negatively regulates seed oil content and provide new insights into the genetic mechanisms driving soybean seed composition and evolution.
{"title":"An α/β-Hydrolase GmABHD6 Controls Seed Oil Content and Yield in Soybean","authors":"Kaixin Yu, Limin Hu, Bo Sun, Zehao Chen, Xu Li, Shiyu Zhang, Huilin Tian, Wenjing Zhang, Shimin Ren, Xue Han, Chang Xu, Siming Wei, Mingliang Yang, Fanjiang Kong, Qingshan Chen, Zhaoming Qi","doi":"10.1111/pbi.70466","DOIUrl":"https://doi.org/10.1111/pbi.70466","url":null,"abstract":"Seed oil content is a key trait in soybean, yet its genetic basis remains largely unresolved. Here, we identify <i>GmABHD6</i>, an α/β-hydrolase domain-containing gene, as the causal gene underlying a major quantitative trait locus (QTL) for seed oil content on chromosome 16. Through fine-mapping in a chromosome segment substitution population derived from <i>Glycine max</i> (SN14) and wild <i>Glycine soja</i> (ZYD00006), we fine-mapped the QTL to a 150-kb interval. Functional analyses reveal that overexpression of <i>GmABHD6</i> significantly reduces seed oil content, whereas gene editing to truncate <i>GmABHD6</i> led to increased oil accumulation. These modifications also alter seed protein levels, seed size and 100-seed weight, indicating pleiotropic effects on seed composition and yield traits. Integrated transcriptomic and metabolomic profiling of transgenic lines highlighted widespread changes in lipid metabolism and energy pathways associated with <i>GmABHD6</i> activity. Further functional characterisation indicates that the ERF transcription factor GmERF physically binds to the <i>GmABHD6</i> promoter region to regulate its expression. Population genetic analyses show strong signatures of selection at the <i>GmABHD6</i> locus, suggesting that favourable alleles are enriched during soybean domestication and improvement. Our findings establish <i>GmABHD6</i> as a domestication-related gene that negatively regulates seed oil content and provide new insights into the genetic mechanisms driving soybean seed composition and evolution.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"7 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145532044","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}
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}
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