Low temperature is a critical abiotic stress that imposes major constraints on the sustainable development of the fruit tree industry. Although exogenous dopamine has been shown to enhance cold tolerance in plants, its molecular mechanisms in apple ( Malus domestica ) remain poorly understood. In this study, we systematically investigated the role of dopamine in cold stress using exogenous dopamine application, overexpression (OE), and RNA interference (RNAi) of the MdTYDC (a key enzyme in dopamine biosynthesis). Our findings demonstrate that dopamine enhances cold resistance in apple through multiple mechanisms, including reducing reactive oxygen species accumulation, improving photosynthesis and stomatal function, promoting anthocyanin biosynthesis, and upregulating CBF genes. Molecular genetic analyses further revealed that MdICE1, a central transcriptional regulator, directly binds to cis‐regulatory elements in the MdTYDC promoter, thereby activating its transcription. Notably, we identified another bHLH transcription factor, MdFAMA, which interacts with MdICE1 and facilitates its binding to the MdTYDC promoter. This interaction amplifies dopamine biosynthesis and strengthens cold resistance. Moreover, exogenous dopamine treatment synergistically induced MdICE1 and MdFAMA expression, forming a positive feedback loop. This feedback mechanism establishes a hierarchical amplification of signalling, further reinforcing tolerance to low temperatures. Collectively, this study elucidates, for the first time, the molecular framework through which the MdICE1/MdFAMA ‐MdTYDC regulatory module orchestrates dopamine‐mediated cold tolerance in apple, providing novel insights into stress adaptation in perennial fruit crops.
{"title":"The MdICE1 / MdFAMA ‐ MdTYDC Transcriptional Module Confers Cold Tolerance by Regulating Dopamine Metabolism in Apple","authors":"Kexin Tan, Xinyang Song, Ziyi Xu, Hongzhen Zhu, Ying Zhang, Shuhan Xu, Zhijun Zhang, Pengmin Li, Fengwang Ma, Chao Li","doi":"10.1111/pbi.70544","DOIUrl":"https://doi.org/10.1111/pbi.70544","url":null,"abstract":"Low temperature is a critical abiotic stress that imposes major constraints on the sustainable development of the fruit tree industry. Although exogenous dopamine has been shown to enhance cold tolerance in plants, its molecular mechanisms in apple ( <jats:styled-content style=\"fixed-case\"> <jats:italic>Malus domestica</jats:italic> </jats:styled-content> ) remain poorly understood. In this study, we systematically investigated the role of dopamine in cold stress using exogenous dopamine application, overexpression (OE), and RNA interference (RNAi) of the <jats:italic>MdTYDC</jats:italic> (a key enzyme in dopamine biosynthesis). Our findings demonstrate that dopamine enhances cold resistance in apple through multiple mechanisms, including reducing reactive oxygen species accumulation, improving photosynthesis and stomatal function, promoting anthocyanin biosynthesis, and upregulating <jats:italic>CBF</jats:italic> genes. Molecular genetic analyses further revealed that MdICE1, a central transcriptional regulator, directly binds to cis‐regulatory elements in the <jats:italic>MdTYDC</jats:italic> promoter, thereby activating its transcription. Notably, we identified another bHLH transcription factor, MdFAMA, which interacts with MdICE1 and facilitates its binding to the <jats:italic>MdTYDC</jats:italic> promoter. This interaction amplifies dopamine biosynthesis and strengthens cold resistance. Moreover, exogenous dopamine treatment synergistically induced <jats:italic>MdICE1</jats:italic> and <jats:italic>MdFAMA</jats:italic> expression, forming a positive feedback loop. This feedback mechanism establishes a hierarchical amplification of signalling, further reinforcing tolerance to low temperatures. Collectively, this study elucidates, for the first time, the molecular framework through which the MdICE1/MdFAMA <jats:italic>‐MdTYDC</jats:italic> regulatory module orchestrates dopamine‐mediated cold tolerance in apple, providing novel insights into stress adaptation in perennial fruit crops.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"17 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145986492","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}
Jongbu Lim, Keunhwa Kim, Jung Heo, Seung Mo Seo, Sungjun Choung, Hyeonjin Kim, Yuri Choi, Kyungsun Park, Hyejung Yun, Dongha Kim, Eun Song Lee, Junwoo Lee, Smita Mirsyad Warsadiharja, Saet Buyl Lee, Sunkyu Han, Soon Ju Park, Sang‐Gyu Kim
Diosgenin is a key starting material for the synthesis of steroidal drugs, such as corticosteroids and sex hormones. While the primary commercial source of diosgenin is the tubers of Dioscorea spp., identifying alternative plant hosts capable of diosgenin biosynthesis could enhance its production. In this study, we present Solanum nigrum , a widely distributed species of the Solanum genus, as a novel platform for diosgenin production. S. nigrum naturally accumulates high concentrations of steroidal glycoalkaloids (SGAs) with a closed F‐ring (spirostanol type) in green fruits and steroidal saponins (STSs) with an open F‐ring (furostanol type) in leaves. Both classes originate from cholesterol and share the early oxidation steps, followed by specific reactions that redirect distinct metabolic fluxes. In SGAs, the CYP450 enzyme SnGAME4 oxidises C26‐OH to an aldehyde, enabling subsequent transamination. In STSs, SnGAME25, a 3β‐hydroxysteroid dehydrogenase/isomerase, initiates the reduction reactions at the C5 double bond. Disruption of these two genes shifted the metabolic profiles from the native SGAs and STSs toward furostanol‐type proto‐diosgenin glycosides. However, these open F‐ring structures yield low diosgenin levels during acid hydrolysis. To overcome this limitation, we identified endogenous furostanol glycoside 26‐ O ‐β‐glucosidases and employed spontaneous fermentation to convert the furostanol structure to the spirostanol structure. Altogether, S. nigrum green fruits yielded diosgenin up to 1% of dry weight. In addition, we engineered S. nigrum to increase fruit number in combination with the SnGAME4 mutation. These results establish S. nigrum as a promising and scalable host for diosgenin production.
{"title":"Rewiring Steroidal Metabolic Pathways for Diosgenin Production in Solanum nigrum","authors":"Jongbu Lim, Keunhwa Kim, Jung Heo, Seung Mo Seo, Sungjun Choung, Hyeonjin Kim, Yuri Choi, Kyungsun Park, Hyejung Yun, Dongha Kim, Eun Song Lee, Junwoo Lee, Smita Mirsyad Warsadiharja, Saet Buyl Lee, Sunkyu Han, Soon Ju Park, Sang‐Gyu Kim","doi":"10.1111/pbi.70551","DOIUrl":"https://doi.org/10.1111/pbi.70551","url":null,"abstract":"Diosgenin is a key starting material for the synthesis of steroidal drugs, such as corticosteroids and sex hormones. While the primary commercial source of diosgenin is the tubers of <jats:italic>Dioscorea</jats:italic> spp., identifying alternative plant hosts capable of diosgenin biosynthesis could enhance its production. In this study, we present <jats:italic>Solanum nigrum</jats:italic> , a widely distributed species of the <jats:italic>Solanum</jats:italic> genus, as a novel platform for diosgenin production. <jats:italic>S. nigrum</jats:italic> naturally accumulates high concentrations of steroidal glycoalkaloids (SGAs) with a closed F‐ring (spirostanol type) in green fruits and steroidal saponins (STSs) with an open F‐ring (furostanol type) in leaves. Both classes originate from cholesterol and share the early oxidation steps, followed by specific reactions that redirect distinct metabolic fluxes. In SGAs, the CYP450 enzyme SnGAME4 oxidises C26‐OH to an aldehyde, enabling subsequent transamination. In STSs, SnGAME25, a 3β‐hydroxysteroid dehydrogenase/isomerase, initiates the reduction reactions at the C5 double bond. Disruption of these two genes shifted the metabolic profiles from the native SGAs and STSs toward furostanol‐type proto‐diosgenin glycosides. However, these open F‐ring structures yield low diosgenin levels during acid hydrolysis. To overcome this limitation, we identified endogenous furostanol glycoside 26‐ <jats:italic>O</jats:italic> ‐β‐glucosidases and employed spontaneous fermentation to convert the furostanol structure to the spirostanol structure. Altogether, <jats:italic>S. nigrum</jats:italic> green fruits yielded diosgenin up to 1% of dry weight. In addition, we engineered <jats:italic>S. nigrum</jats:italic> to increase fruit number in combination with the <jats:italic>SnGAME4</jats:italic> mutation. These results establish <jats:italic>S. nigrum</jats:italic> as a promising and scalable host for diosgenin production.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"47 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145986541","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}
Jakob Weber Böhlen, Alexander Beesley, Sebastian F. Beyer, Patrick Schwinges, Alina E. Maas, Holger Schultheiss, Uwe Conrath, Caspar J. G. Langenbach
{"title":"AtMYB72 as a Biotechnological Tool to Overcome Phenylpropanoid Substrate Limitation and Enhance Coumarin Biosynthesis in Plants","authors":"Jakob Weber Böhlen, Alexander Beesley, Sebastian F. Beyer, Patrick Schwinges, Alina E. Maas, Holger Schultheiss, Uwe Conrath, Caspar J. G. Langenbach","doi":"10.1111/pbi.70503","DOIUrl":"https://doi.org/10.1111/pbi.70503","url":null,"abstract":"","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"9 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145986494","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}
Yifeng Shi,Yue Xu,Hai Li,Meng Fu,Xu Liu,Yuxiang Li,Xiaoping Hu
As global warming continues, rising temperatures significantly alter the interactions between wheat and the stripe rust pathogen Puccinia striiformis f. sp. tritici (Pst). Utilising high-temperature all-stage (HTAS) resistance to Pst is a novel strategy for breeding climate and disease resilient wheat cultivars. Cysteine-rich receptor-like kinases (CRKs) are involved in massive transduction pathways upon perception of biotic and abiotic stresses in plants. Here, we identify a CRK subfamily gene, TaCRK6, from Xiaoyan 6 (XY6), a wheat cultivar possessing non-race-specific and durable HTAS resistance to stripe rust. The expression of TaCRK6 concurrently responds to both Pst inoculation and the relatively high temperature treatment. Silencing TaCRK6 significantly attenuated HTAS resistance to Pst in XY6. Furthermore, overexpression of TaCRK6 in susceptible wheat cultivar Fielder exhibited a resistant phenotype with reduced Pst sporulation and increased necrosis. TaCRK6 interacts with and primarily phosphorylates the cytoplasmic kinase TaRLCK185 with the threonine residue at position 248. Notably, the MAPK signalling cascades, positioned downstream of TaRLCK185, are proved to participate in activating HTAS resistance in XY6. TaRLCK185 transduces the MAPK cascade signals by interacting with and primarily phosphorylating the serine residue of TaMAPKKK1 at position 132. TaCRK6-mediated phosphorylation of T248 alters the conformation of TaRLCK185, which in turn promotes its interaction with TaMAPKKK1, ultimately leading to activation of the downstream TaMAPKKK1-TaMAPKK9-TaMAPK6 cascade. Moreover, the TaCRK6-TaRLCK185-TaMAPKs module regulates the biosynthesis of salicylic acid (SA). These results indicate a TaCRK6-TaRLCK185-TaMAPKs module that transduces dual stress signals, coupling with the SA pathway initiation to ultimately activate HTAS resistance against Pst in XY6.
{"title":"The Wheat CRK-RLCK-MAPKs Signalling Module Confers High-Temperature All-Stage Resistance to Stripe Rust.","authors":"Yifeng Shi,Yue Xu,Hai Li,Meng Fu,Xu Liu,Yuxiang Li,Xiaoping Hu","doi":"10.1111/pbi.70537","DOIUrl":"https://doi.org/10.1111/pbi.70537","url":null,"abstract":"As global warming continues, rising temperatures significantly alter the interactions between wheat and the stripe rust pathogen Puccinia striiformis f. sp. tritici (Pst). Utilising high-temperature all-stage (HTAS) resistance to Pst is a novel strategy for breeding climate and disease resilient wheat cultivars. Cysteine-rich receptor-like kinases (CRKs) are involved in massive transduction pathways upon perception of biotic and abiotic stresses in plants. Here, we identify a CRK subfamily gene, TaCRK6, from Xiaoyan 6 (XY6), a wheat cultivar possessing non-race-specific and durable HTAS resistance to stripe rust. The expression of TaCRK6 concurrently responds to both Pst inoculation and the relatively high temperature treatment. Silencing TaCRK6 significantly attenuated HTAS resistance to Pst in XY6. Furthermore, overexpression of TaCRK6 in susceptible wheat cultivar Fielder exhibited a resistant phenotype with reduced Pst sporulation and increased necrosis. TaCRK6 interacts with and primarily phosphorylates the cytoplasmic kinase TaRLCK185 with the threonine residue at position 248. Notably, the MAPK signalling cascades, positioned downstream of TaRLCK185, are proved to participate in activating HTAS resistance in XY6. TaRLCK185 transduces the MAPK cascade signals by interacting with and primarily phosphorylating the serine residue of TaMAPKKK1 at position 132. TaCRK6-mediated phosphorylation of T248 alters the conformation of TaRLCK185, which in turn promotes its interaction with TaMAPKKK1, ultimately leading to activation of the downstream TaMAPKKK1-TaMAPKK9-TaMAPK6 cascade. Moreover, the TaCRK6-TaRLCK185-TaMAPKs module regulates the biosynthesis of salicylic acid (SA). These results indicate a TaCRK6-TaRLCK185-TaMAPKs module that transduces dual stress signals, coupling with the SA pathway initiation to ultimately activate HTAS resistance against Pst in XY6.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"4 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145961380","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}
<p>The idea of using plants as biofactories to produce medically valuable proteins was formalized some 35 years ago by Andy Hiatt, Robert Cafferkey and Katherine Bowdish in a paper reporting the successful expression of mammalian antibodies in tobacco (Hiatt et al. <span>1989</span>), soon followed by Peter Sijmons and co-workers describing the correct processing of human serum albumin in potato (Sijmons et al. <span>1990</span>). These seminal papers, followed by thousands of others over the following decades, have paved the way for the emergence of ‘plant molecular farming’, a now thriving discipline of plant biotechnology dedicated to the heterologous production of valuable proteins and organics in plant systems. Plant-based expression platforms have been developed to produce a wide array of valuable recombinant products, including vaccine antigens, therapeutic antibodies, bioactive proteins and (poly)peptides (Chaudhary et al. <span>2024</span>; Eidenberger et al. <span>2023</span>; Stander et al. <span>2022</span>), and, more recently, small organic chemicals (Golubova et al. <span>2024</span>; Liu et al. <span>2023</span>). The commercialization of plant-made recombinant protein products and the expected approval of several others for human use in the coming years have strengthened the position of plant molecular farming as a mature, viable option for the heterologous production of useful protein and organic products (Washida et al. <span>2025</span>; Anon. <span>2022</span>).</p><p>One reason for the success of plant molecular farming is the development of transient expression platforms involving leaf infiltration with engineered <i>Agrobacterium tumefaciens</i> (now referred to as <i>Rhizobium radiobacter</i>) (Akher et al. <span>2025</span>) harboring expression vectors optimized for plant-based protein production. Originally used to quickly screen for functional promoter sequences and gene constructs, transient expression in agroinfiltrated leaves has since developed into the fastest and most convenient production platform for plant-made (bio)pharmaceuticals. This approach relies on the ability of the <i>A. tumefaciens</i> Ti plasmid to transfer a transcriptionally competent segment of DNA into the plant's host cell, that is then directed to the nucleus for heterologous expression. Several transient expression systems have been devised over the years, that enable the production of milligram quantities of recombinant proteins within a few days in a handful of plants.</p><p>Most of these systems make use of the wild tobacco relative <i>Nicotiana benthamiana</i> as an expression host (Bally et al. <span>2018</span>). Widely adopted as an experimental model to elucidate plant-pathogen interactions, this plant rapidly generates leaf biomass and is easily amenable to agroinfiltration for transgene introduction and high-level expression. In recent years, <i>N. benthamiana</i> has become the most widely used host for transient protein productio
大约35年前，Andy Hiatt、Robert Cafferkey和Katherine Bowdish在一篇报道在烟草中成功表达哺乳动物抗体的论文中正式提出了利用植物作为生物工厂生产具有医学价值的蛋白质的想法（Hiatt et al. 1989），随后Peter Sijmons和同事描述了在马铃薯中正确处理人类血清白蛋白（Sijmons et al. 1990）。在接下来的几十年里，这些开创性的论文以及成千上万的其他论文为“植物分子农业”的出现铺平了道路，这是一个现在蓬勃发展的植物生物技术学科，致力于在植物系统中异种生产有价值的蛋白质和有机物。基于植物的表达平台已被开发用于生产各种有价值的重组产品，包括疫苗抗原、治疗性抗体、生物活性蛋白和（多）肽（Chaudhary等人，2024；Eidenberger等人，2023；Stander等人，2022），以及最近的小有机化学品（Golubova等人，2024；Liu等人，2023）。植物重组蛋白产品的商业化以及未来几年预计将批准其他几种用于人类的产品，加强了植物分子农业作为有用蛋白质和有机产品异种生产的成熟、可行选择的地位（Washida et al. 2025; Anon. 2022）。植物分子农业成功的一个原因是瞬时表达平台的发展，包括用工程农杆菌（现在称为放射根瘤菌）（Akher et al. 2025）渗透叶片，其中含有针对植物蛋白生产优化的表达载体。最初用于快速筛选功能启动子序列和基因结构，而今在农渗叶片中的瞬时表达已发展成为最快、最方便的植物合成（生物）药物生产平台。这种方法依赖于瘤化芽孢杆菌Ti质粒将具有转录能力的DNA片段转移到植物的宿主细胞中，然后将其定向到细胞核中进行异源表达。多年来，已经设计出了几种瞬时表达系统，可以在几天内在少数植物中产生毫克级的重组蛋白。这些系统大多数使用野生烟草的亲缘本烟作为表达宿主（Bally et al. 2018）。该植物被广泛用作阐明植物与病原体相互作用的实验模型，其叶片生物量产生迅速，易于通过农业渗透进行转基因导入和高水平表达。近年来，N. benthamiana已成为最广泛使用的瞬时蛋白生产宿主，每周（如果不是每天）都会发表几篇研究论文，报道在该植物中成功表达有价值的重组蛋白或重建代谢途径以产生有用的代谢物。在过去的20年里，benthamiana在植物生物技术中日益重要，PBJ在分子农业平台的发展中发挥了重要作用，基于此，本期特刊提供了该领域当前发展前沿的最新文章。权威、深刻的综述首先讨论了与benthamiana表达平台相关的特定主题，从寄主植物对农杆菌感染的反应和转基因表达的当前策略，到重组蛋白的设计、环境控制和重组蛋白产品的下游加工。一流的初级研究论文随后会呈现关于新生物制药生产的原始数据报告，提出一种提高重组蛋白产量和/或叶组织质量的新方法，或描述一种独特的代谢工程策略来生产临床感兴趣的有机化合物。本期收录论文28篇，其中综述6篇，简要交流6篇，全文研究16篇。还将在期刊网站上发表一个补充的虚拟问题，包括2025年在常规问题上发表的相同的28篇论文和相关论文，以及仍在制作中的其他一些论文，包括一篇额外的评论。一旦完成，特刊（可通过虚拟特刊获得）将包括来自世界各地近40个实验室的约40篇高质量论文。我们要对所有积极参与这项集体工作的贡献者表示感谢和赞赏，这项工作可以说是迄今为止完全致力于benthamiana生物工厂的最全面的出版工作之一，如果不是最全面的话。 我们还要向在此过程中征聘的100多名外部审稿人表示最深切的感谢，他们的专业知识和辛勤工作直接促进了最终结果的总体质量。最后，我们感谢Wiley团队在项目过程中给予的宝贵帮助，以及PBJ主编jonathan Napier教授从开始到结束的宝贵建议和持续支持。多米尼克·米肖，PBJ高级编辑。Stephen J. Streatfield， PBJ副主编。数据共享不适用于本文，因为在当前研究中没有生成或分析数据集。
{"title":"The Nicotiana benthamiana Biofactory","authors":"Dominique Michaud, Stephen J. Streatfield","doi":"10.1111/pbi.70547","DOIUrl":"10.1111/pbi.70547","url":null,"abstract":"<p>The idea of using plants as biofactories to produce medically valuable proteins was formalized some 35 years ago by Andy Hiatt, Robert Cafferkey and Katherine Bowdish in a paper reporting the successful expression of mammalian antibodies in tobacco (Hiatt et al. <span>1989</span>), soon followed by Peter Sijmons and co-workers describing the correct processing of human serum albumin in potato (Sijmons et al. <span>1990</span>). These seminal papers, followed by thousands of others over the following decades, have paved the way for the emergence of ‘plant molecular farming’, a now thriving discipline of plant biotechnology dedicated to the heterologous production of valuable proteins and organics in plant systems. Plant-based expression platforms have been developed to produce a wide array of valuable recombinant products, including vaccine antigens, therapeutic antibodies, bioactive proteins and (poly)peptides (Chaudhary et al. <span>2024</span>; Eidenberger et al. <span>2023</span>; Stander et al. <span>2022</span>), and, more recently, small organic chemicals (Golubova et al. <span>2024</span>; Liu et al. <span>2023</span>). The commercialization of plant-made recombinant protein products and the expected approval of several others for human use in the coming years have strengthened the position of plant molecular farming as a mature, viable option for the heterologous production of useful protein and organic products (Washida et al. <span>2025</span>; Anon. <span>2022</span>).</p><p>One reason for the success of plant molecular farming is the development of transient expression platforms involving leaf infiltration with engineered <i>Agrobacterium tumefaciens</i> (now referred to as <i>Rhizobium radiobacter</i>) (Akher et al. <span>2025</span>) harboring expression vectors optimized for plant-based protein production. Originally used to quickly screen for functional promoter sequences and gene constructs, transient expression in agroinfiltrated leaves has since developed into the fastest and most convenient production platform for plant-made (bio)pharmaceuticals. This approach relies on the ability of the <i>A. tumefaciens</i> Ti plasmid to transfer a transcriptionally competent segment of DNA into the plant's host cell, that is then directed to the nucleus for heterologous expression. Several transient expression systems have been devised over the years, that enable the production of milligram quantities of recombinant proteins within a few days in a handful of plants.</p><p>Most of these systems make use of the wild tobacco relative <i>Nicotiana benthamiana</i> as an expression host (Bally et al. <span>2018</span>). Widely adopted as an experimental model to elucidate plant-pathogen interactions, this plant rapidly generates leaf biomass and is easily amenable to agroinfiltration for transgene introduction and high-level expression. In recent years, <i>N. benthamiana</i> has become the most widely used host for transient protein productio","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 1","pages":"3-4"},"PeriodicalIF":10.5,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70547","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145962151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Potato (Solanum tuberosum L.) is a globally important food crop with considerable nutritional and economic value. Heat stress significantly inhibits potato plant growth and tuber development, constraining the sustainable development of the potato industry. Currently, studies on the cellular-level mechanisms underlying heat adaptation in potato remain relatively scarce. In this study, single-nucleus RNA sequencing was employed to construct single-cell transcriptomic maps of potato leaves under normal and heat stress conditions, yielding 77 344 high-quality nuclei and identifying six major cell types. The results indicated that epidermal cells represented the key cell type in heat-stress response, exhibiting the highest number of differentially expressed genes, whereas vascular cells were positioned in the transition zone of the pseudo-time trajectory and may have been involved in cell differentiation processes. By integrating bulk RNA-seq data, a heat stress response co-expression network was constructed, identifying 12 core transcription factors, with StPIF4 appearing repeatedly. Experimental validation confirmed that heat stress strongly induced StPIF4 expression. Functional studies demonstrated that StPIF4 significantly enhanced potato heat tolerance by improving reactive oxygen species scavenging capacity. This study provided cellular-level insights into the mechanisms underlying potato adaptation to heat stress.
{"title":"Transcription Profiling of Potato Leaves in Response to Heat Stress at Single-Cell Resolution.","authors":"Shiqi Wen,Ke Wang,Wenqian Liang,Rongrong Liu,Zihan Li,Xinlong Chen,Yan Li,Dianqiu Lv,Hongju Jian","doi":"10.1111/pbi.70546","DOIUrl":"https://doi.org/10.1111/pbi.70546","url":null,"abstract":"Potato (Solanum tuberosum L.) is a globally important food crop with considerable nutritional and economic value. Heat stress significantly inhibits potato plant growth and tuber development, constraining the sustainable development of the potato industry. Currently, studies on the cellular-level mechanisms underlying heat adaptation in potato remain relatively scarce. In this study, single-nucleus RNA sequencing was employed to construct single-cell transcriptomic maps of potato leaves under normal and heat stress conditions, yielding 77 344 high-quality nuclei and identifying six major cell types. The results indicated that epidermal cells represented the key cell type in heat-stress response, exhibiting the highest number of differentially expressed genes, whereas vascular cells were positioned in the transition zone of the pseudo-time trajectory and may have been involved in cell differentiation processes. By integrating bulk RNA-seq data, a heat stress response co-expression network was constructed, identifying 12 core transcription factors, with StPIF4 appearing repeatedly. Experimental validation confirmed that heat stress strongly induced StPIF4 expression. Functional studies demonstrated that StPIF4 significantly enhanced potato heat tolerance by improving reactive oxygen species scavenging capacity. This study provided cellular-level insights into the mechanisms underlying potato adaptation to heat stress.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"49 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145955945","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}
Cuifang Zhu,Hongjun Yu,Caili Zhao,Hongyang Wu,Xiaoyang Wan,Tao Lu,Yang Li,Weijie Jiang,Qiang Li
In the context of declining arable land, the development of plant architectures that maximise the use of finite resources is crucial for addressing food security. This study collected yield data, along with aboveground and root traits, from 263 cucumber varieties. Machine learning models and scenario simulations were utilised with the goal of identifying a high-yielding cucumber architecture suitable for greenhouse cultivation. Our findings indicate that cucumber yields can be predicted using aboveground and root phenotypes, such as the position of the first female flower node, leaf width, stem diameter, and root angle, with the combination of GBDT and SVM algorithms yielding the most accurate results (R2 = 0.6155, RMSE = 0.2601). Analysis of 157 464 phenotypic combinations revealed antagonistic interactions between robust aboveground structures and fine root systems, and synergistic interactions between slender aboveground parts and broad root systems. Yields were up to 20% higher in phenotypes that combined a compact, robust aboveground structure with a narrow yet larger-diameter and shallower root system, reflecting additive effects rather than synergistic ones. Additionally, this study proposes a reference range for high-yielding phenotypes. Overall, this research provides a theoretical foundation for optimising cucumber plant structures under greenhouse environments by predicting yields and investigating phenotypic interactions through modelling.
{"title":"Machine Learning-Driven Construction of High-Yielding Cucumber Plant Architectures in Greenhouse Environments.","authors":"Cuifang Zhu,Hongjun Yu,Caili Zhao,Hongyang Wu,Xiaoyang Wan,Tao Lu,Yang Li,Weijie Jiang,Qiang Li","doi":"10.1111/pbi.70539","DOIUrl":"https://doi.org/10.1111/pbi.70539","url":null,"abstract":"In the context of declining arable land, the development of plant architectures that maximise the use of finite resources is crucial for addressing food security. This study collected yield data, along with aboveground and root traits, from 263 cucumber varieties. Machine learning models and scenario simulations were utilised with the goal of identifying a high-yielding cucumber architecture suitable for greenhouse cultivation. Our findings indicate that cucumber yields can be predicted using aboveground and root phenotypes, such as the position of the first female flower node, leaf width, stem diameter, and root angle, with the combination of GBDT and SVM algorithms yielding the most accurate results (R2 = 0.6155, RMSE = 0.2601). Analysis of 157 464 phenotypic combinations revealed antagonistic interactions between robust aboveground structures and fine root systems, and synergistic interactions between slender aboveground parts and broad root systems. Yields were up to 20% higher in phenotypes that combined a compact, robust aboveground structure with a narrow yet larger-diameter and shallower root system, reflecting additive effects rather than synergistic ones. Additionally, this study proposes a reference range for high-yielding phenotypes. Overall, this research provides a theoretical foundation for optimising cucumber plant structures under greenhouse environments by predicting yields and investigating phenotypic interactions through modelling.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"46 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145949544","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}
Aoyu Ling,Yijia Jin,Yufei Xia,Shenxiu Jiang,Jianghai Shu,Xiaotong Hu,Kang Du,Pingdong Zhang,Xiangyang Kang
Base excision repair (BER) is a critical pathway for repairing damaged DNA bases in cells; however, the mechanisms of protein recruitment and interaction in this pathway remain largely unexplored in higher plants. In this study, we used '84K' poplar (Populus alba × P. glandulosa) as the experimental system and applied a low concentration of 5-aminouracil (5-AU) to induce DNA base lesions. Through transcriptome analysis and weighted gene co-expression network analysis (WGCNA), we identified two key BER-responsive genes: the DNA glycosylase family gene PagDMG6341 and the DNA polymerase δ subunit PagPOLD4. PagDMG6341 was significantly upregulated during the arrest phase of 5-AU treatment, whereas PagPOLD4 expression peaked during the subsequent release phase. RNA interference (RNAi) lines for each gene resulted in impaired growth and increased susceptibility to 5-AU in '84K' poplar, supporting their functional roles in DNA repair and development. To further investigate their potential interaction network, we performed yeast two-hybrid (Y2H) screening, AlphaFold3-based structural modelling, confirmatory Y2H, bimolecular fluorescence complementation (BiFC) assays, and luciferase complementation imaging (LCI) assays. These experiments demonstrated that a Transducin/WD40-repeat-like scaffold protein (PagWD40) interacts independently with both PagDMG6341 and PagPOLD4. The yeast three-hybrid (Y3H) assay further showed that PagWD40 functions as a molecular scaffold, linking PagDMG6341 and PagPOLD4 to form a functional complex. This study reveals a new mechanism in which PagWD40 functions as a scaffold protein linking a DNA glycosylase with DNA polymerase δ in the plant BER pathway, thereby providing new insights into the organisation of plant DNA damage repair networks.
碱基切除修复（BER）是修复细胞中受损DNA碱基的重要途径；然而，在高等植物中，这一途径中的蛋白质募集和相互作用机制仍未得到充分研究。本研究以84K杨树(Populus alba × P。并应用低浓度的5-氨基尿嘧啶（5-AU）诱导DNA碱基病变。通过转录组分析和加权基因共表达网络分析（WGCNA），我们确定了两个关键的ber响应基因：DNA糖基化酶家族基因PagDMG6341和DNA聚合酶δ亚基PagPOLD4。PagDMG6341在5-AU处理的阻滞期显著上调，而PagPOLD4的表达在随后的释放期达到峰值。每个基因的RNA干扰（RNAi）系导致84K杨树生长受损和对5-AU的易感性增加，支持它们在DNA修复和发育中的功能作用。为了进一步研究它们潜在的相互作用网络，我们进行了酵母双杂交（Y2H）筛选、基于alphafold3的结构建模、验证性Y2H、双分子荧光互补（BiFC）检测和荧光素酶互补成像（LCI）检测。这些实验表明，转导蛋白/ wd40 -重复样支架蛋白（PagWD40）与PagDMG6341和PagPOLD4独立相互作用。酵母三杂交（Y3H）实验进一步表明，PagWD40作为分子支架，连接PagDMG6341和PagPOLD4形成功能复合物。该研究揭示了植物BER通路中PagWD40作为连接DNA糖基酶和DNA聚合酶δ的支架蛋白的新机制，从而为植物DNA损伤修复网络的组织提供了新的见解。
{"title":"The PagDMG6341-PagWD40-PagPOLD4 Module Coordinates Base Excision Repair in '84K' Poplar (Populus alba × P. glandulosa).","authors":"Aoyu Ling,Yijia Jin,Yufei Xia,Shenxiu Jiang,Jianghai Shu,Xiaotong Hu,Kang Du,Pingdong Zhang,Xiangyang Kang","doi":"10.1111/pbi.70543","DOIUrl":"https://doi.org/10.1111/pbi.70543","url":null,"abstract":"Base excision repair (BER) is a critical pathway for repairing damaged DNA bases in cells; however, the mechanisms of protein recruitment and interaction in this pathway remain largely unexplored in higher plants. In this study, we used '84K' poplar (Populus alba × P. glandulosa) as the experimental system and applied a low concentration of 5-aminouracil (5-AU) to induce DNA base lesions. Through transcriptome analysis and weighted gene co-expression network analysis (WGCNA), we identified two key BER-responsive genes: the DNA glycosylase family gene PagDMG6341 and the DNA polymerase δ subunit PagPOLD4. PagDMG6341 was significantly upregulated during the arrest phase of 5-AU treatment, whereas PagPOLD4 expression peaked during the subsequent release phase. RNA interference (RNAi) lines for each gene resulted in impaired growth and increased susceptibility to 5-AU in '84K' poplar, supporting their functional roles in DNA repair and development. To further investigate their potential interaction network, we performed yeast two-hybrid (Y2H) screening, AlphaFold3-based structural modelling, confirmatory Y2H, bimolecular fluorescence complementation (BiFC) assays, and luciferase complementation imaging (LCI) assays. These experiments demonstrated that a Transducin/WD40-repeat-like scaffold protein (PagWD40) interacts independently with both PagDMG6341 and PagPOLD4. The yeast three-hybrid (Y3H) assay further showed that PagWD40 functions as a molecular scaffold, linking PagDMG6341 and PagPOLD4 to form a functional complex. This study reveals a new mechanism in which PagWD40 functions as a scaffold protein linking a DNA glycosylase with DNA polymerase δ in the plant BER pathway, thereby providing new insights into the organisation of plant DNA damage repair networks.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"30 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145949545","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}
Crops are continually challenged by biotic stresses, including fungal, bacterial and viral pathogens and insect pests, which cause substantial yield and quality losses worldwide. WRKY transcription factors constitute a plant-specific and functionally diverse family that is central to immune regulation. Recent advances in genomic resources and multi-omics approaches have accelerated the identification and functional characterisation of WRKYs in crops. This review summarises the structural features and classification of WRKY genes and their genome-wide distribution across crop species. It also synthesises WRKY-centred regulatory modules that mediate resistance to major classes of biotic stress. In antifungal defence, WRKYs reinforce pattern- and effector-triggered immunity, modulate protein stability and reprogramme secondary metabolism. In antibacterial immunity, they link bacterial perception to cell wall remodelling and hormone and redox signalling. WRKYs also activate PR gene expression, cell wall fortification, RNA interference and programmed cell death to combat oomycete and viral pathogens and insect pests. Overall, WRKYs function as context-dependent transcriptional hubs. They integrate immune signalling with hormonal crosstalk, remodel defence gene networks, and redirect secondary metabolism, thereby shaping resistance outcomes under biotic stress. The review examines WRKY-mediated defence-growth trade-offs and explores opportunities to harness WRKY-centred networks for breeding and engineering broad-spectrum, durable disease and pest resistance. It also highlights how integrating multi-omics with precision genome editing, synthetic biology, gene-drive technologies and artificial intelligence could establish WRKYs as central molecular targets for improving crop resilience and performance.
{"title":"WRKY Transcription Factors: Integral Regulators of Defence Responses to Biotic Stress in Crops.","authors":"Dongjiao Wang,Ruize Zhang,Wenhui Zou,Yuanyuan Zhang,Wanying Zhao,Tingting Sun,Qibin Wu,Zheng Qing Fu,Youxiong Que","doi":"10.1111/pbi.70542","DOIUrl":"https://doi.org/10.1111/pbi.70542","url":null,"abstract":"Crops are continually challenged by biotic stresses, including fungal, bacterial and viral pathogens and insect pests, which cause substantial yield and quality losses worldwide. WRKY transcription factors constitute a plant-specific and functionally diverse family that is central to immune regulation. Recent advances in genomic resources and multi-omics approaches have accelerated the identification and functional characterisation of WRKYs in crops. This review summarises the structural features and classification of WRKY genes and their genome-wide distribution across crop species. It also synthesises WRKY-centred regulatory modules that mediate resistance to major classes of biotic stress. In antifungal defence, WRKYs reinforce pattern- and effector-triggered immunity, modulate protein stability and reprogramme secondary metabolism. In antibacterial immunity, they link bacterial perception to cell wall remodelling and hormone and redox signalling. WRKYs also activate PR gene expression, cell wall fortification, RNA interference and programmed cell death to combat oomycete and viral pathogens and insect pests. Overall, WRKYs function as context-dependent transcriptional hubs. They integrate immune signalling with hormonal crosstalk, remodel defence gene networks, and redirect secondary metabolism, thereby shaping resistance outcomes under biotic stress. The review examines WRKY-mediated defence-growth trade-offs and explores opportunities to harness WRKY-centred networks for breeding and engineering broad-spectrum, durable disease and pest resistance. It also highlights how integrating multi-omics with precision genome editing, synthetic biology, gene-drive technologies and artificial intelligence could establish WRKYs as central molecular targets for improving crop resilience and performance.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"81 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145949547","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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