Autophagy and the ubiquitin/26S proteasome system (UPS) play critical roles in the immune defence of the host against pathogen invasion. As a countermeasure, pathogens deploy effector proteins to subvert or hijack autophagy and UPS processes. However, it is unclear whether and how a single pathogen effector coordinately modulates both proteolytic systems. Here, we identified a RING finger E3 ligase of Citrus sinensis , CsRHY1A, that directly interacts with SDE4405, an effector protein from Candidatus Liberibacter asiaticus ( C Las), the causal agent of citrus Huanglongbing (HLB). CsRHY1A ubiquitinated SDE4405 at Lys87 and Lys92, causing SDE4405 degradation via the 26S proteasome. Furthermore, SDE4405 targeted the ubiquitin‐associated (UBA) domain of the autophagic receptor NEIGHBOR OF BRCA1 (CsNBR1) and competitively disrupted CsRHY1A‐mediated degradation by decreasing the ubiquitination of SDE4405. Lys87 and Lys92 of SDE4405 were required for its interactions with CsRHY1A and CsNBR1 and were essential for CsNBR1‐dependent stabilisation of SDE4405. SDE4405 also inhibited the binding of CsNBR1 to CsATG8s, suppressing CsNBR1‐mediated selective autophagic degradation of C Las effector protein SDE1. These findings reveal the sophisticated strategy of bacteria to counteract both autophagy and proteasome‐dependent degradation, providing opportunities for developing HLB‐resistant citrus varieties.
自噬和泛素/26S蛋白酶体系统（UPS）在宿主抵御病原体入侵的免疫防御中起着至关重要的作用。作为对策，病原体部署效应蛋白来破坏或劫持自噬和UPS过程。然而，目前尚不清楚单一病原体效应是否以及如何协调调节这两个蛋白水解系统。本研究中，我们鉴定了柑橘的RING finger E3连接酶CsRHY1A，该连接酶可直接与柑橘黄龙冰（HLB）病原菌亚洲解放候选菌（Candidatus Liberibacter asiaticus, C Las）的效应蛋白SDE4405相互作用。CsRHY1A在Lys87和Lys92位点泛素化SDE4405，导致SDE4405通过26S蛋白酶体降解。此外，SDE4405靶向BRCA1自噬受体邻居（CsNBR1）的泛素相关（UBA）结构域，并通过降低SDE4405的泛素化，竞争性地破坏了CsRHY1A介导的降解。SDE4405的Lys87和Lys92是其与CsRHY1A和CsNBR1相互作用所必需的，并且是SDE4405的CsNBR1依赖性稳定所必需的。SDE4405还抑制CsNBR1与CsATG8s的结合，抑制CsNBR1介导的C Las效应蛋白SDE1的选择性自噬降解。这些发现揭示了细菌对抗自噬和蛋白酶体依赖性降解的复杂策略，为开发抗HLB柑橘品种提供了机会。
{"title":"A Bacterial Effector Hijacks NBR1 to Modulate Both Autophagy and Ubiquitination‐Mediated Degradation That Promotes Bacterial Infection","authors":"Yaqian Shi, Fang Fang, Xuejin Cui, Hongwei Shi, Zaiyu Yang, Xueyi Li, Changyong Zhou, Xuefeng Wang","doi":"10.1111/pbi.70509","DOIUrl":"https://doi.org/10.1111/pbi.70509","url":null,"abstract":"Autophagy and the ubiquitin/26S proteasome system (UPS) play critical roles in the immune defence of the host against pathogen invasion. As a countermeasure, pathogens deploy effector proteins to subvert or hijack autophagy and UPS processes. However, it is unclear whether and how a single pathogen effector coordinately modulates both proteolytic systems. Here, we identified a RING finger E3 ligase of <jats:styled-content style=\"fixed-case\"> <jats:italic>Citrus sinensis</jats:italic> </jats:styled-content> , CsRHY1A, that directly interacts with SDE4405, an effector protein from <jats:italic>Candidatus</jats:italic> Liberibacter asiaticus ( <jats:italic>C</jats:italic> Las), the causal agent of citrus Huanglongbing (HLB). CsRHY1A ubiquitinated SDE4405 at Lys87 and Lys92, causing SDE4405 degradation via the 26S proteasome. Furthermore, SDE4405 targeted the ubiquitin‐associated (UBA) domain of the autophagic receptor NEIGHBOR OF BRCA1 (CsNBR1) and competitively disrupted CsRHY1A‐mediated degradation by decreasing the ubiquitination of SDE4405. Lys87 and Lys92 of SDE4405 were required for its interactions with CsRHY1A and CsNBR1 and were essential for CsNBR1‐dependent stabilisation of SDE4405. SDE4405 also inhibited the binding of CsNBR1 to CsATG8s, suppressing CsNBR1‐mediated selective autophagic degradation of <jats:italic>C</jats:italic> Las effector protein SDE1. These findings reveal the sophisticated strategy of bacteria to counteract both autophagy and proteasome‐dependent degradation, providing opportunities for developing HLB‐resistant citrus varieties.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"4 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145765467","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}
Hiroyuki Kajiura, Kana Yamamoto, Ryo Misaki, Kazuhito Fujiyama
Plants are promising next‐generation hosts for recombinant protein production; however, major challenges remain with regard to enhancing the efficiency of downstream processing, particularly in the removal of cellular residues and purification of the expressed proteins. Strategies to overcome these limitations include targeting expressed recombinant proteins within a specific organelle or directing their secretion into the extracellular space, thereby facilitating purification by collecting the target matrix. In this study, we focused on protein secretion mechanisms and identified two pathogenesis‐related proteins, glucan endo‐1,3‐β‐glucosidase (GN) and chitinase 8 (Chi8), which accumulated in the apoplast washing fluid following Agrobacterium infiltration of Nicotiana benthamiana leaves. Both proteins contained signal peptides (SPs), SP GN and SP Chi8 , respectively. Although the intracellular accumulation of GFP was comparable regardless of the expression level, fusion with either SP GN or SP Chi8 resulted in GFP accumulation within the apoplast. In contrast, in N. benthamiana , a mammalian‐derived SP was less effective in facilitating GFP secretion than the plant‐derived SPs. Additionally, replacing the SP of the mammalian‐derived protein β‐glucocerebrosidase (GCase) with SP GN or SP Chi8 enhanced the secretion of GCase into the apoplast, indicating their applicability in protein production. Moreover, SP GN and SP Chi8 directed the expressed proteins into the culture medium of N. benthamiana suspension cells. These results indicate that SP GN and SP Chi8 function as effective secretion signals and highlight the potential application of endogenous SPs for enhancing recombinant protein production in plants.
{"title":"Protein‐Derived Signal Peptides Induced by Agrobacterium Infection Promote the Secretion of Recombinant Proteins in Nicotiana benthamiana","authors":"Hiroyuki Kajiura, Kana Yamamoto, Ryo Misaki, Kazuhito Fujiyama","doi":"10.1111/pbi.70498","DOIUrl":"https://doi.org/10.1111/pbi.70498","url":null,"abstract":"Plants are promising next‐generation hosts for recombinant protein production; however, major challenges remain with regard to enhancing the efficiency of downstream processing, particularly in the removal of cellular residues and purification of the expressed proteins. Strategies to overcome these limitations include targeting expressed recombinant proteins within a specific organelle or directing their secretion into the extracellular space, thereby facilitating purification by collecting the target matrix. In this study, we focused on protein secretion mechanisms and identified two pathogenesis‐related proteins, glucan endo‐1,3‐β‐glucosidase (GN) and chitinase 8 (Chi8), which accumulated in the apoplast washing fluid following <jats:italic>Agrobacterium</jats:italic> infiltration of <jats:italic>Nicotiana benthamiana</jats:italic> leaves. Both proteins contained signal peptides (SPs), SP <jats:sub>GN</jats:sub> and SP <jats:sub>Chi8</jats:sub> , respectively. Although the intracellular accumulation of GFP was comparable regardless of the expression level, fusion with either SP <jats:sub>GN</jats:sub> or SP <jats:sub>Chi8</jats:sub> resulted in GFP accumulation within the apoplast. In contrast, in <jats:italic>N. benthamiana</jats:italic> , a mammalian‐derived SP was less effective in facilitating GFP secretion than the plant‐derived SPs. Additionally, replacing the SP of the mammalian‐derived protein β‐glucocerebrosidase (GCase) with SP <jats:sub>GN</jats:sub> or SP <jats:sub>Chi8</jats:sub> enhanced the secretion of GCase into the apoplast, indicating their applicability in protein production. Moreover, SP <jats:sub>GN</jats:sub> and SP <jats:sub>Chi8</jats:sub> directed the expressed proteins into the culture medium of <jats:italic>N. benthamiana</jats:italic> suspension cells. These results indicate that SP <jats:sub>GN</jats:sub> and SP <jats:sub>Chi8</jats:sub> function as effective secretion signals and highlight the potential application of endogenous SPs for enhancing recombinant protein production in plants.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"16 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145759512","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}
Tobamoviruses establish viral replication organelles (VROs) on the host endoplasmic reticulum (ER) for their replication, a process demanding substantial different types of lipids. However, how viruses efficiently transfer these lipids from other compartments like chloroplasts remains incompletely understood. Fibrillin (FBN) proteins are primarily localised to chloroplasts and are intimately linked with lipid metabolism and stress responses in chloroplasts. Here, we report that NbLIP1, a light‐induced FBN1 family protein in Nicotiana benthamiana , directly interacts with the coat protein (CP) of rehmannia mosaic virus (ReMV) and other related tobamoviruses (TMV, YoMV). Upon viral infection, this interaction leads to the relocalisation of NbLIP1 from chloroplasts to ER‐proximal viral replication sites. Functional assays demonstrated that overexpression of NbLIP1 significantly enhanced viral replication, viral protein accumulation, and VRO formation, while silencing NbLIP1 had the opposite effects. These findings unveil a novel viral infection mechanism whereby the viral CP hijacks the host lipid transfer protein NbLIP1 and recruits it to viral replication factories to promote viral replication, potentially by modulating lipid supply or the microenvironment remodelling at the replication sites. This study not only elucidates the role of NbLIP1 as a novel pro‐viral host factor but also provides new insights into how viruses exploit host resources across cellular compartments, suggesting NbLIP1 as a potential antiviral target.
{"title":"Tobamoviruses CP Proteins Hijack Light‐Induced Protein ( NbLIP1 ) to Promote Viral Replication by Facilitating VRO Formation","authors":"Haoyu Chen, Mingjie Wu, Xiao Guo, Hongmei Xu, Chenwei Feng, Duxuan Liu, Jing Hua, Yanhong Hua, Zhen He, Peter Moffett, Kun Zhang","doi":"10.1111/pbi.70497","DOIUrl":"https://doi.org/10.1111/pbi.70497","url":null,"abstract":"Tobamoviruses establish viral replication organelles (VROs) on the host endoplasmic reticulum (ER) for their replication, a process demanding substantial different types of lipids. However, how viruses efficiently transfer these lipids from other compartments like chloroplasts remains incompletely understood. Fibrillin (FBN) proteins are primarily localised to chloroplasts and are intimately linked with lipid metabolism and stress responses in chloroplasts. Here, we report that NbLIP1, a light‐induced FBN1 family protein in <jats:italic>Nicotiana benthamiana</jats:italic> , directly interacts with the coat protein (CP) of rehmannia mosaic virus (ReMV) and other related tobamoviruses (TMV, YoMV). Upon viral infection, this interaction leads to the relocalisation of NbLIP1 from chloroplasts to ER‐proximal viral replication sites. Functional assays demonstrated that overexpression of NbLIP1 significantly enhanced viral replication, viral protein accumulation, and VRO formation, while silencing NbLIP1 had the opposite effects. These findings unveil a novel viral infection mechanism whereby the viral CP hijacks the host lipid transfer protein NbLIP1 and recruits it to viral replication factories to promote viral replication, potentially by modulating lipid supply or the microenvironment remodelling at the replication sites. This study not only elucidates the role of NbLIP1 as a novel pro‐viral host factor but also provides new insights into how viruses exploit host resources across cellular compartments, suggesting NbLIP1 as a potential antiviral target.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"10 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145759501","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}
Fabio G. Moratti, Chiara Lonoce, Stephan Obst, Xenia Kroop, Daniel Karcher, Stephanie Ruf, Ralph Bock
{"title":"A Set of Intein‐Split Selectable Marker Genes for Efficient Co‐Transformation","authors":"Fabio G. Moratti, Chiara Lonoce, Stephan Obst, Xenia Kroop, Daniel Karcher, Stephanie Ruf, Ralph Bock","doi":"10.1111/pbi.70502","DOIUrl":"https://doi.org/10.1111/pbi.70502","url":null,"abstract":"","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"77 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145759502","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}
Abiotic stresses severely constrain soybean productivity worldwide. Here we demonstrate that gmeif2b5 (eukaryotic initiation factors) mutants confer dual stress tolerance through coordinated mechanisms. Integrative RNA-Seq and protein interaction analyses revealed that gmeif2b5 mutants increase lignin deposition to increase drought resilience and balanced Na+/K+ homeostasis to enhance salt tolerance; GmeIF2B5 physically interacts with GmPRX4, a plant heme peroxidase; GmPRX4 overexpression increases drought and salt resistance in soybean; GmeIF2B5 plays the predominant role in the GmeIF2B5-GmPRX4 module, and double mutants exhibiting synergistic stress tolerance improvements. Our work uncovers a 'GmeIF2B5-GmPRX4 regulatory axis' that: mobilises lignin-based structural fortification for enhanced drought resistance and orchestrates ionic equilibrium for increased salt tolerance. This study pioneers the role of eIF2B genes in soybean stress adaptation, establishing a multi-tiered regulatory node for precision molecular design of stress-resilient crops.
{"title":"A GmeIF2B5-GmPRX4 Regulatory Axis Divergently Governs Drought-Lignin and Salt-Ion Homeostasis in Soybean.","authors":"Juan Liu,Yanzhong Huang,Xiaowan Fang,Han Gou,Huidong Xuan,Sushuang Deng,Lu Li,Yanjia Wang,Xiushuai Wang,Ling Gan,Nannan Zhang,Haoran Luo,Yaolan Bai,Qin Liu,Han Xing,Jinming Zhao,Na Guo","doi":"10.1111/pbi.70507","DOIUrl":"https://doi.org/10.1111/pbi.70507","url":null,"abstract":"Abiotic stresses severely constrain soybean productivity worldwide. Here we demonstrate that gmeif2b5 (eukaryotic initiation factors) mutants confer dual stress tolerance through coordinated mechanisms. Integrative RNA-Seq and protein interaction analyses revealed that gmeif2b5 mutants increase lignin deposition to increase drought resilience and balanced Na+/K+ homeostasis to enhance salt tolerance; GmeIF2B5 physically interacts with GmPRX4, a plant heme peroxidase; GmPRX4 overexpression increases drought and salt resistance in soybean; GmeIF2B5 plays the predominant role in the GmeIF2B5-GmPRX4 module, and double mutants exhibiting synergistic stress tolerance improvements. Our work uncovers a 'GmeIF2B5-GmPRX4 regulatory axis' that: mobilises lignin-based structural fortification for enhanced drought resistance and orchestrates ionic equilibrium for increased salt tolerance. This study pioneers the role of eIF2B genes in soybean stress adaptation, establishing a multi-tiered regulatory node for precision molecular design of stress-resilient crops.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"230 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145752798","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}
Jieyu Chen, Chuanhezi Quan, Yang Zhao, Imani L. D. S. Kalumith, Zhangjun Fei, Leon V. Kochian, William J. Lucas, Byung‐Kook Ham
Phosphorus (P) is an essential macronutrient for various biological processes in plant growth. Modern agricultural science has advanced the knowledge of regulatory mechanisms underlying phosphorus starvation responses (PSRs), aiming to develop phosphate‐efficient crops with sustainable production under reduced Pi fertilizer application. However, information regarding coordinated shoot and root adaptations in response to combined nutrient stresses is limited. This study investigated the role of Phloem Phosphate Stress Repressed 1 (PPSR1) in modulating PSRs and other nutrient deficiency adaptations. The Arabidopsis functional homologue of Cucumis sativus PPSR1 (CsPPSR1), designated AtPPSR1, was identified. AtPPSR1 encodes a glycine‐rich domain‐containing protein, and its ectopic expression confers enhanced growth performance to plants. Transcriptomic analyses revealed AtPPSR1 as a regulatory mediator of PSRs, photosynthesis, and root development. AtPPSR1 interacted with PHOSPHATE STARVATION RESPONSE 1 (PHR1) to regulate PHR1‐target genes for adaptive root development in response to Pi‐starvation stress. Additionally, AtPPSR1 was graft‐transmissible, and shoot‐borne AtPPSR1 played a role in restoring the root phenotype of the ppsr1 mutant. Physiological analyses revealed that enhanced AtPPSR1 expression enabled resilience to nitrogen (N) and potassium (K)‐starvation, as well as to Pi‐deficiency. Furthermore, we identified homologues of CsPPSR1 and AtPPSR1 in Brassica napus (canola), which displayed similar expression patterns in response to Pi‐starvation stress. Overexpression of PPSR1 , identified from Arabidopsis, cucumber, and canola, improved growth performance and seed production in canola under N‐, Pi‐, or K‐deficient conditions, within the controlled environment. These findings provide novel insights into PPSR1‐mediated molecular coordination to enhance plant resilience to mineral nutrient deficiency.
{"title":"PPSR1 Protein Functions as an Important Regulator to Enhance Plant Growth Performance Under N, P, and K Deficient Stress Conditions","authors":"Jieyu Chen, Chuanhezi Quan, Yang Zhao, Imani L. D. S. Kalumith, Zhangjun Fei, Leon V. Kochian, William J. Lucas, Byung‐Kook Ham","doi":"10.1111/pbi.70496","DOIUrl":"https://doi.org/10.1111/pbi.70496","url":null,"abstract":"Phosphorus (P) is an essential macronutrient for various biological processes in plant growth. Modern agricultural science has advanced the knowledge of regulatory mechanisms underlying phosphorus starvation responses (PSRs), aiming to develop phosphate‐efficient crops with sustainable production under reduced Pi fertilizer application. However, information regarding coordinated shoot and root adaptations in response to combined nutrient stresses is limited. This study investigated the role of Phloem Phosphate Stress Repressed 1 (PPSR1) in modulating PSRs and other nutrient deficiency adaptations. The Arabidopsis functional homologue of <jats:styled-content style=\"fixed-case\"> <jats:italic>Cucumis sativus</jats:italic> </jats:styled-content> PPSR1 (CsPPSR1), designated AtPPSR1, was identified. AtPPSR1 encodes a glycine‐rich domain‐containing protein, and its ectopic expression confers enhanced growth performance to plants. Transcriptomic analyses revealed AtPPSR1 as a regulatory mediator of PSRs, photosynthesis, and root development. AtPPSR1 interacted with PHOSPHATE STARVATION RESPONSE 1 (PHR1) to regulate PHR1‐target genes for adaptive root development in response to Pi‐starvation stress. Additionally, AtPPSR1 was graft‐transmissible, and shoot‐borne AtPPSR1 played a role in restoring the root phenotype of the <jats:italic>ppsr1</jats:italic> mutant. Physiological analyses revealed that enhanced <jats:italic>AtPPSR1</jats:italic> expression enabled resilience to nitrogen (N) and potassium (K)‐starvation, as well as to Pi‐deficiency. Furthermore, we identified homologues of CsPPSR1 and AtPPSR1 in <jats:styled-content style=\"fixed-case\"> <jats:italic>Brassica napus</jats:italic> </jats:styled-content> (canola), which displayed similar expression patterns in response to Pi‐starvation stress. Overexpression of <jats:italic>PPSR1</jats:italic> , identified from Arabidopsis, cucumber, and canola, improved growth performance and seed production in canola under N‐, Pi‐, or K‐deficient conditions, within the controlled environment. These findings provide novel insights into PPSR1‐mediated molecular coordination to enhance plant resilience to mineral nutrient deficiency.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"12 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145731165","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}
Daniel Lunn, Alayna Trejo, Baskaran Kannan, Amandine Germon, Alistair Leverett, Tom E. Clemente, Fredy Altpeter, Andrew D. B. Leakey
Stomata are microscopic pores that regulate the exchange of CO 2 and water vapour, making them a major target for engineering plants with improved intrinsic water use efficiency (iWUE). Proof‐of‐concept studies have demonstrated the potential to increase iWUE by reducing stomatal density (SD) and stomatal conductance (g sw ) by ubiquitously expressing EPIDERMAL PATTERNING FACTOR (EPF) family genes. However, unwanted effects on leaf, stem and reproductive traits are often observed when EPFs are misexpressed in this fashion. We sought to test if these effects result from pleiotropy and to identify a targeted promoter that can circumvent the side effects while retaining the desired reduction in SD. A previously reported synthetic EPF (EPF syn ) was expressed in sugarcane ( Saccharum spp.) using two putatively tissue‐specific promoters from Brachypodium distachyon (BdCESA7p and BdSPCH2p) and a ubiquitous control from Zea mays (ZmUBI4p). BdSPCH2p control reduced SD to statistically equivalent levels as ZmUBI4p on the abaxial (23%) and adaxial (23%) leaf surfaces. ZmUB4p and BdCESA7p induce expression in four tissue types often associated with pleiotropic effects in EPF‐expressing low SD plants. Transgenic plants carrying either the BdCESA7p or ZmUBI4p EPF syn cassettes displayed leaf chlorosis, reduced leaf nitrogen and chlorophyll content, and altered stem architecture. However, transgenic events harboring the BdSPCH2p EPF syn cassette restricted EPF syn expression to the stomatal development zone and leaf nodal tissues and produced transgenic plants without the associated pleiotropic effects. These results represent an important step toward engineering low‐SD crops since they show that targeted gene expression can engineer stomatal patterning without impairing agronomically important traits.
{"title":"Brachypodium SPEECHLESS2 Promoter Drives Expression of a Synthetic EPF to Reduce Stomatal Density in Sugarcane Without Pleiotropic Effects","authors":"Daniel Lunn, Alayna Trejo, Baskaran Kannan, Amandine Germon, Alistair Leverett, Tom E. Clemente, Fredy Altpeter, Andrew D. B. Leakey","doi":"10.1111/pbi.70495","DOIUrl":"https://doi.org/10.1111/pbi.70495","url":null,"abstract":"Stomata are microscopic pores that regulate the exchange of CO <jats:sub>2</jats:sub> and water vapour, making them a major target for engineering plants with improved intrinsic water use efficiency (iWUE). Proof‐of‐concept studies have demonstrated the potential to increase iWUE by reducing stomatal density (SD) and stomatal conductance (g <jats:sub>sw</jats:sub> ) by ubiquitously expressing EPIDERMAL PATTERNING FACTOR (EPF) family genes. However, unwanted effects on leaf, stem and reproductive traits are often observed when EPFs are misexpressed in this fashion. We sought to test if these effects result from pleiotropy and to identify a targeted promoter that can circumvent the side effects while retaining the desired reduction in SD. A previously reported synthetic EPF (EPF <jats:sub>syn</jats:sub> ) was expressed in sugarcane ( <jats:italic>Saccharum</jats:italic> spp.) using two putatively tissue‐specific promoters from <jats:italic>Brachypodium distachyon</jats:italic> (BdCESA7p and BdSPCH2p) and a ubiquitous control from <jats:italic>Zea mays</jats:italic> (ZmUBI4p). BdSPCH2p control reduced SD to statistically equivalent levels as ZmUBI4p on the abaxial (23%) and adaxial (23%) leaf surfaces. ZmUB4p and BdCESA7p induce expression in four tissue types often associated with pleiotropic effects in EPF‐expressing low SD plants. Transgenic plants carrying either the BdCESA7p or ZmUBI4p EPF <jats:sub>syn</jats:sub> cassettes displayed leaf chlorosis, reduced leaf nitrogen and chlorophyll content, and altered stem architecture. However, transgenic events harboring the BdSPCH2p EPF <jats:sub>syn</jats:sub> cassette restricted EPF <jats:sub>syn</jats:sub> expression to the stomatal development zone and leaf nodal tissues and produced transgenic plants without the associated pleiotropic effects. These results represent an important step toward engineering low‐SD crops since they show that targeted gene expression can engineer stomatal patterning without impairing agronomically important traits.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"145 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145731166","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}
Yinghui Dong, Fei Su, Hailin Xie, Sinan Zhang, Meng Jia, Chunyu Zou, Mugui Wang, Jian‐Kang Zhu
{"title":"A Rice Endogenous Small RNA ‐Binding Protein Improves Prime Editing for Precise Sequence Insertion and Replacement","authors":"Yinghui Dong, Fei Su, Hailin Xie, Sinan Zhang, Meng Jia, Chunyu Zou, Mugui Wang, Jian‐Kang Zhu","doi":"10.1111/pbi.70468","DOIUrl":"https://doi.org/10.1111/pbi.70468","url":null,"abstract":"","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"7 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145731161","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}
Colletotrichum spp., hemibiotrophic fungal pathogens, threaten global strawberry production. Jasmonate (JA) regulates plant‐ Colletotrichum interactions, but its mechanisms remain unclear. Here we demonstrate that both exogenous methyl jasmonate (MeJA) treatment and elevated endogenous MeJA levels increase strawberry susceptibility to anthracnose. Two key JA biosynthesis genes, FveAOS2 and FveAOC3 , were identified as contributors to Colletotrichum ‐induced susceptibility. Further analysis revealed that the FveSnRK2.1–FveWRKY50 phosphorylation module functions as an important molecular switch in regulating disease susceptibility. Specifically, Colletotrichum infection or MeJA application activates FveSnRK2.1, which phosphorylates FveWRKY50 at serine residue 88 (S88). This phosphorylation enhances the stability and transcriptional activity of FveWRKY50, leading to increased expression of FveAOS2 and FveAOC3 , higher MeJA accumulation and enhanced susceptibility. Notably, the strawberry JASMONATE‐ZIM DOMAIN (JAZ) protein FveJAZ5 suppresses susceptibility by directly interacting with FveWRKY50, thereby preventing its interaction with FveSnRK2.1 and inhibiting the activation of FveAOS2 and FveAOC3 . Upon pathogen attack or MeJA signalling, FveJAZ5 is degraded, thereby releasing FveWRKY50 from suppression. The study elucidates a Colletotrichum ‐induced ‘JA signaling – JA biosynthesis’ positive feedback loop that drives strawberry susceptibility. Knocking out FveWRKY50 and overexpressing FveJAZ5 generated anthracnose‐resistant germplasms. These findings deepen understanding of plant‐ Colletotrichum interactions and provide genes for resistant strawberry breeding.
{"title":"Jasmonate Modulates Strawberry Susceptibility to Anthracnose by Activating SnRK2.1 to Regulate the WRKY50‐JAZ5 Module","authors":"Chuang Liu, Zhen Liu, Xia Li, Yating Chen, Ronghui Sun, Peijie Li, Qianqian Feng, Yuanhua Wang, Jie Ren, Qian Li, Bingbing Li","doi":"10.1111/pbi.70492","DOIUrl":"https://doi.org/10.1111/pbi.70492","url":null,"abstract":"<jats:italic>Colletotrichum</jats:italic> spp., hemibiotrophic fungal pathogens, threaten global strawberry production. Jasmonate (JA) regulates plant‐ <jats:italic>Colletotrichum</jats:italic> interactions, but its mechanisms remain unclear. Here we demonstrate that both exogenous methyl jasmonate (MeJA) treatment and elevated endogenous MeJA levels increase strawberry susceptibility to anthracnose. Two key JA biosynthesis genes, <jats:italic>FveAOS2</jats:italic> and <jats:italic>FveAOC3</jats:italic> , were identified as contributors to <jats:italic>Colletotrichum</jats:italic> ‐induced susceptibility. Further analysis revealed that the FveSnRK2.1–FveWRKY50 phosphorylation module functions as an important molecular switch in regulating disease susceptibility. Specifically, <jats:italic>Colletotrichum</jats:italic> infection or MeJA application activates FveSnRK2.1, which phosphorylates FveWRKY50 at serine residue 88 (S88). This phosphorylation enhances the stability and transcriptional activity of FveWRKY50, leading to increased expression of <jats:italic>FveAOS2</jats:italic> and <jats:italic>FveAOC3</jats:italic> , higher MeJA accumulation and enhanced susceptibility. Notably, the strawberry JASMONATE‐ZIM DOMAIN (JAZ) protein FveJAZ5 suppresses susceptibility by directly interacting with FveWRKY50, thereby preventing its interaction with FveSnRK2.1 and inhibiting the activation of <jats:italic>FveAOS2</jats:italic> and <jats:italic>FveAOC3</jats:italic> . Upon pathogen attack or MeJA signalling, FveJAZ5 is degraded, thereby releasing FveWRKY50 from suppression. The study elucidates a <jats:italic>Colletotrichum</jats:italic> ‐induced ‘JA signaling – JA biosynthesis’ positive feedback loop that drives strawberry susceptibility. Knocking out <jats:italic>FveWRKY50</jats:italic> and overexpressing <jats:italic>FveJAZ5</jats:italic> generated anthracnose‐resistant germplasms. These findings deepen understanding of plant‐ <jats:italic>Colletotrichum</jats:italic> interactions and provide genes for resistant strawberry breeding.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"144 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145731164","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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