Cytoplasmic male sterility (CMS) and restorer of fertility (Rf) are important traits in F1 hybrid breeding. However, the CMS/Rf system has not been used in tomatoes because of the limited resources of Rf lines. In this study, we performed mutagenesis in tomato CMS seeds and successfully obtained 13 suppressor mutants with pollen fertility. Using bulked segregant analysis and whole-genome sequencing for each suppressor mutant, we detected mutations associated with fertility restoration in the nuclear-encoded gene for the mitochondrial RNA polymerase termed SlRPOTm in four independent mutants created through mutagenesis. Furthermore, we found that the loss of function of SlRPOTm was associated with fertility restoration in the tomato CMS line. Expression analysis of orf137, a tomato CMS-causing gene, revealed that reduced expression of orf137 was associated with fertility restoration in tomato CMS. In addition, F1 plants carrying mutations in SlRPOTm were generated, and tomato fruit formation was comparable to that of normal F1 plants. This study demonstrates for the first time that the loss of function of mitochondrial RNA polymerase contributes to fertility restoration in CMS lines. Furthermore, it is possible to replace various tomato varieties with Rf lines using genome editing technology, which will promote tomato F1 breeding in the future.
{"title":"Artificial Mutations in the Nuclear Gene Encoding Mitochondrial RNA Polymerase Restore Pollen Fertility in Cytoplasmic Male Sterile Tomato.","authors":"Kosuke Kuwabara, Rika Nakajima, Van Bosstraeten Alexis Gaetan, Kentaro Ezura, Kinya Toriyama, Tohru Ariizumi, Kenta Shirasawa","doi":"10.1111/pbi.70417","DOIUrl":"https://doi.org/10.1111/pbi.70417","url":null,"abstract":"<p><p>Cytoplasmic male sterility (CMS) and restorer of fertility (Rf) are important traits in F<sub>1</sub> hybrid breeding. However, the CMS/Rf system has not been used in tomatoes because of the limited resources of Rf lines. In this study, we performed mutagenesis in tomato CMS seeds and successfully obtained 13 suppressor mutants with pollen fertility. Using bulked segregant analysis and whole-genome sequencing for each suppressor mutant, we detected mutations associated with fertility restoration in the nuclear-encoded gene for the mitochondrial RNA polymerase termed SlRPOTm in four independent mutants created through mutagenesis. Furthermore, we found that the loss of function of SlRPOTm was associated with fertility restoration in the tomato CMS line. Expression analysis of orf137, a tomato CMS-causing gene, revealed that reduced expression of orf137 was associated with fertility restoration in tomato CMS. In addition, F<sub>1</sub> plants carrying mutations in SlRPOTm were generated, and tomato fruit formation was comparable to that of normal F<sub>1</sub> plants. This study demonstrates for the first time that the loss of function of mitochondrial RNA polymerase contributes to fertility restoration in CMS lines. Furthermore, it is possible to replace various tomato varieties with Rf lines using genome editing technology, which will promote tomato F<sub>1</sub> breeding in the future.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2025-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145342370","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}
Woody biomass is a promising source of fermentable sugars for biofuels and bio-based chemicals, but its industrial use is limited by the costly biorefinery process. A viable strategy to reduce costs involves enhancing both biomass processability and the generation of high-value co-products. Here, we report the implementation of a synthetic metabolic pathway in Populus tremula × P. alba to produce 2-pyrone-4,6-dicarboxylic acid (PDC), a key building block for biodegradable plastics and high-performance materials. This artificial pathway-comprising microbial genes AroG, QsuB, PmdA, PmdB, and PmdC-enabled de novo PDC production in the stems of transgenic poplar. Pathway expression also induced substantial metabolic reprogramming and altered cell wall composition. These include the hyperaccumulation of simple phenolics like protocatechuic acid (PCA) and vanillic acid (VA), alongside reduced levels of p-hydroxybenzoic acid. A large portion of VA was ester-linked to cell wall lignin, while PCA was incorporated into the lignin backbone, forming novel benzodioxane units; concurrently, lignin in transgenic plants exhibited a drastic reduction in guaiacyl- and syringyl-units, with a notable increase in p-hydroxyphenyl-units. Hemicellulose content, particularly xylan, was also significantly increased. Moreover, expression of the PDC-pathway led to the formation of novel VA-derived suberin aromatics, enhancing suberization in bark and roots and improving salt stress tolerance. These changes led to improved saccharification efficiency, with up to 25% more glucose and 2.5 times xylose released from woody biomass. These results demonstrate the metabolic flexibility of poplar and highlight its potential for engineering cost-effective, stress-resilient bioenergy crops with enhanced biorefinery traits.
{"title":"Engineering 2-Pyrone-4,6-Dicarboxylic Acid Production Reveals Metabolic Plasticity of Poplar.","authors":"Nidhi Dwivedi,Pingping Ji,Yang Tian,Dasmeet Kaur,Vijaya Kumar Reddy Vulavala,Yuki Tobimatsu,Aymerick Eudes,Chang-Jun Liu","doi":"10.1111/pbi.70414","DOIUrl":"https://doi.org/10.1111/pbi.70414","url":null,"abstract":"Woody biomass is a promising source of fermentable sugars for biofuels and bio-based chemicals, but its industrial use is limited by the costly biorefinery process. A viable strategy to reduce costs involves enhancing both biomass processability and the generation of high-value co-products. Here, we report the implementation of a synthetic metabolic pathway in Populus tremula × P. alba to produce 2-pyrone-4,6-dicarboxylic acid (PDC), a key building block for biodegradable plastics and high-performance materials. This artificial pathway-comprising microbial genes AroG, QsuB, PmdA, PmdB, and PmdC-enabled de novo PDC production in the stems of transgenic poplar. Pathway expression also induced substantial metabolic reprogramming and altered cell wall composition. These include the hyperaccumulation of simple phenolics like protocatechuic acid (PCA) and vanillic acid (VA), alongside reduced levels of p-hydroxybenzoic acid. A large portion of VA was ester-linked to cell wall lignin, while PCA was incorporated into the lignin backbone, forming novel benzodioxane units; concurrently, lignin in transgenic plants exhibited a drastic reduction in guaiacyl- and syringyl-units, with a notable increase in p-hydroxyphenyl-units. Hemicellulose content, particularly xylan, was also significantly increased. Moreover, expression of the PDC-pathway led to the formation of novel VA-derived suberin aromatics, enhancing suberization in bark and roots and improving salt stress tolerance. These changes led to improved saccharification efficiency, with up to 25% more glucose and 2.5 times xylose released from woody biomass. These results demonstrate the metabolic flexibility of poplar and highlight its potential for engineering cost-effective, stress-resilient bioenergy crops with enhanced biorefinery traits.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"54 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145339091","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}
Na-Young Choi,Min-Ha Kim,Hyun-A Jang,Seung-Won Pyo,Kong-Young Park,Hyoshin Lee,Eun-Kyung Bae,Jae-Heung Ko
Narrow or upright branch angles in shoots and leaves lead to columnar, upright-growing tree architectures, as observed in various tree species such as Lombardy poplar (Populus nigra var. italica). However, the genetic mechanism underlying this unique growth habit in Lombardy poplar has not yet been elucidated. In this study, we identified a nonsense mutation in the PnTAC1-1 gene of Lombardy poplar, an ortholog of the rice TILLER ANGLE CONTROL 1 (TAC1) gene known to regulate branch angles. To confirm the functional role of TAC1 in regulating tree architecture, we generated transgenic hybrid poplar (Populus alba × Populus glandulosa, clone BH) with targeted mutations in TAC1 homologues using CRISPR/Cas9 gene editing. The resulting TAC1-CRISPR hybrid poplars exhibited a stable upright branching phenotype closely resembling that of Lombardy poplar, as confirmed by two consecutive years of living modified organism (LMO) field trials. Anatomical analysis revealed increased cell elongation specifically in the lower petiole region and significantly enhanced gravitropic responses in TAC1-CRISPR hybrid poplars compared to wild-type BH clones. RNA sequencing analysis further demonstrated that TAC1 disruption triggered extensive transcriptomic reprogramming of axillary meristem, notably altering hormonal and photomorphogenic signalling pathways, which redirected auxin accumulation toward the abaxial region and increased gibberellin biosynthesis, ultimately promoting upright growth. This research uncovers the genetic and molecular mechanisms behind columnar growth in poplar and provides a promising approach for engineering tree architecture to enhance planting density, harvest efficiency and woody biomass productivity.
在不同的树种中，如伦巴第白杨（Populus nigra var. italica），可以观察到枝条的狭窄或垂直角度导致柱状、直立生长的树木结构。然而，伦巴第杨树这种独特生长习性的遗传机制尚未阐明。在这项研究中，我们在伦巴第杨树的PnTAC1-1基因中发现了一个无义突变，该基因是水稻分蘖角度控制1 （TAC1）基因的同源物，已知该基因调节分枝角度。为了证实TAC1在调控树型中的功能作用，我们利用CRISPR/Cas9基因编辑技术，构建了TAC1同源基因靶向突变的转基因杂交杨树（Populus alba × Populus glandulosa，克隆BH）。经连续两年的活改性生物（LMO）田间试验证实，TAC1-CRISPR杂交杨树具有稳定的直立分枝表型，与伦巴第杨树非常相似。解剖分析显示，与野生型BH克隆相比，TAC1-CRISPR杂交杨树的下叶柄区细胞伸长明显增加，向地性反应显著增强。RNA测序分析进一步表明，TAC1的破坏引发了腋窝分生组织广泛的转录组重编程，显著改变了激素和光形态形成信号通路，将生长素的积累重定向到腋窝区域，增加了赤霉素的生物合成，最终促进直立生长。该研究揭示了杨树柱状生长的遗传和分子机制，为提高树木种植密度、收获效率和木材生物量生产力提供了有希望的途径。
{"title":"Elucidating the Genetic Basis of Columnar Upright Architecture in Populus Through CRISPR Disruption of TILLER ANGLE CONTROL1.","authors":"Na-Young Choi,Min-Ha Kim,Hyun-A Jang,Seung-Won Pyo,Kong-Young Park,Hyoshin Lee,Eun-Kyung Bae,Jae-Heung Ko","doi":"10.1111/pbi.70415","DOIUrl":"https://doi.org/10.1111/pbi.70415","url":null,"abstract":"Narrow or upright branch angles in shoots and leaves lead to columnar, upright-growing tree architectures, as observed in various tree species such as Lombardy poplar (Populus nigra var. italica). However, the genetic mechanism underlying this unique growth habit in Lombardy poplar has not yet been elucidated. In this study, we identified a nonsense mutation in the PnTAC1-1 gene of Lombardy poplar, an ortholog of the rice TILLER ANGLE CONTROL 1 (TAC1) gene known to regulate branch angles. To confirm the functional role of TAC1 in regulating tree architecture, we generated transgenic hybrid poplar (Populus alba × Populus glandulosa, clone BH) with targeted mutations in TAC1 homologues using CRISPR/Cas9 gene editing. The resulting TAC1-CRISPR hybrid poplars exhibited a stable upright branching phenotype closely resembling that of Lombardy poplar, as confirmed by two consecutive years of living modified organism (LMO) field trials. Anatomical analysis revealed increased cell elongation specifically in the lower petiole region and significantly enhanced gravitropic responses in TAC1-CRISPR hybrid poplars compared to wild-type BH clones. RNA sequencing analysis further demonstrated that TAC1 disruption triggered extensive transcriptomic reprogramming of axillary meristem, notably altering hormonal and photomorphogenic signalling pathways, which redirected auxin accumulation toward the abaxial region and increased gibberellin biosynthesis, ultimately promoting upright growth. This research uncovers the genetic and molecular mechanisms behind columnar growth in poplar and provides a promising approach for engineering tree architecture to enhance planting density, harvest efficiency and woody biomass productivity.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"78 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145338890","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}
Ty Shitanaka,Yu Wang,Sally Do,Julia Yuson,Samir Kumar Khanal,Krzysztof Zienkiewicz,Zhi-Yan Du
Microalgae are increasingly recognised as powerful platforms for the sustainable production of lipids and terpenoids, with expanding applications in the food, fuel and biomanufacturing industries. In this updated review, we consolidate and critically assess the most recent advances in synthetic biology and metabolic engineering of key microalgal models, including Chlamydomonas reinhardtii, Nannochloropsis spp. and Phaeodactylum tricornutum. We focus on developments that have emerged in the latest waves of research, emphasising novel genetic toolkits that accelerate the Design-Build-Test-Learn (DBTL) cycle, breakthroughs in genome-scale metabolic modelling, and innovative strategies for organelle-targeted biosynthesis of high-value compounds. Recent case studies are compared to highlight trends in successful engineering approaches. By capturing these up-to-date insights, this review outlines the current trajectory of microalgal biotechnology toward scalable, carbon-neutral biofactories for polyunsaturated fatty acids (PUFAs) and diverse terpenoids, reinforcing their role in global sustainability and the circular bioeconomy.
{"title":"Synthetic Biology and Metabolic Engineering of Microalgae for Sustainable Lipid and Terpenoid Production: An Updated Perspective.","authors":"Ty Shitanaka,Yu Wang,Sally Do,Julia Yuson,Samir Kumar Khanal,Krzysztof Zienkiewicz,Zhi-Yan Du","doi":"10.1111/pbi.70405","DOIUrl":"https://doi.org/10.1111/pbi.70405","url":null,"abstract":"Microalgae are increasingly recognised as powerful platforms for the sustainable production of lipids and terpenoids, with expanding applications in the food, fuel and biomanufacturing industries. In this updated review, we consolidate and critically assess the most recent advances in synthetic biology and metabolic engineering of key microalgal models, including Chlamydomonas reinhardtii, Nannochloropsis spp. and Phaeodactylum tricornutum. We focus on developments that have emerged in the latest waves of research, emphasising novel genetic toolkits that accelerate the Design-Build-Test-Learn (DBTL) cycle, breakthroughs in genome-scale metabolic modelling, and innovative strategies for organelle-targeted biosynthesis of high-value compounds. Recent case studies are compared to highlight trends in successful engineering approaches. By capturing these up-to-date insights, this review outlines the current trajectory of microalgal biotechnology toward scalable, carbon-neutral biofactories for polyunsaturated fatty acids (PUFAs) and diverse terpenoids, reinforcing their role in global sustainability and the circular bioeconomy.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"41 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145331616","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}
Ying Liu, Irene Merino, Mareike Gutensohn, Annika I. Johansson, Kalle Johansson, Mariette Andersson, Per Hofvander, Folke Sitbon
Steroidal glycoalkaloids (SGAs) are toxic cholesterol‐derived secondary metabolites present in several Solanaceae species. In potato, tuber SGA levels are for reasons of toxicity of concern in both table and starch cultivars. In the latter, SGAs bind to proteins and fibres in starch production side‐streams and prevent their further uses as food and feed. To enable more sustainable uses of starch by‐products, we have here reduced SGA biosynthesis in a starch potato cultivar using DNA‐free CRISPR/Cas9. Six SGA genes were targeted, encoding enzymes acting either before cholesterol (SMO1‐L, DWF1‐L, DWF7‐L), or after (16DOX, CYP88B1, TAMiso2). Editing efficiencies varied between 20% and 49%, and generated mutants were investigated under greenhouse and field conditions. Target mass‐spectrometric analyses confirmed reduced SGA levels and alterations of sterol metabolism in mutated events. Plant height and tuber yield were reduced in several events, although this was not correlated to low SGA levels. Several knockout mutants had almost SGA‐free leaves and tubers, the latter also under two SGA‐inducing conditions. Similarly, both fibre and protein fractions isolated from side‐streams in the starch production process from mutant tubers had very low SGA levels. By contrast, the corresponding wild‐type SGA levels were almost 10‐fold and, respectively, 40‐fold higher than the recommended upper safe limit. The results demonstrate that glycoalkaloid‐free mutants can be generated and grown with moderate yield reductions under both greenhouse and field conditions. This suggests a potential for sustainable production of high‐value products, e.g., food‐grade protein and fibre, from starch production side‐streams of SGA knockout tubers.
{"title":"Glycoalkaloid‐Free Starch Potatoes Generated by CRISPR/Cas9‐Mediated Mutations of Genes in the Glycoalkaloid Biosynthesis Pathway Enable More Sustainable Uses of By‐Products From Starch Production","authors":"Ying Liu, Irene Merino, Mareike Gutensohn, Annika I. Johansson, Kalle Johansson, Mariette Andersson, Per Hofvander, Folke Sitbon","doi":"10.1111/pbi.70412","DOIUrl":"https://doi.org/10.1111/pbi.70412","url":null,"abstract":"Steroidal glycoalkaloids (SGAs) are toxic cholesterol‐derived secondary metabolites present in several Solanaceae species. In potato, tuber SGA levels are for reasons of toxicity of concern in both table and starch cultivars. In the latter, SGAs bind to proteins and fibres in starch production side‐streams and prevent their further uses as food and feed. To enable more sustainable uses of starch by‐products, we have here reduced SGA biosynthesis in a starch potato cultivar using DNA‐free CRISPR/Cas9. Six SGA genes were targeted, encoding enzymes acting either before cholesterol (<jats:italic>SMO1‐L</jats:italic>, <jats:italic>DWF1‐L</jats:italic>, <jats:italic>DWF7‐L</jats:italic>), or after (<jats:italic>16DOX</jats:italic>, <jats:italic>CYP88B1</jats:italic>, <jats:italic>TAMiso2</jats:italic>). Editing efficiencies varied between 20% and 49%, and generated mutants were investigated under greenhouse and field conditions. Target mass‐spectrometric analyses confirmed reduced SGA levels and alterations of sterol metabolism in mutated events. Plant height and tuber yield were reduced in several events, although this was not correlated to low SGA levels. Several knockout mutants had almost SGA‐free leaves and tubers, the latter also under two SGA‐inducing conditions. Similarly, both fibre and protein fractions isolated from side‐streams in the starch production process from mutant tubers had very low SGA levels. By contrast, the corresponding wild‐type SGA levels were almost 10‐fold and, respectively, 40‐fold higher than the recommended upper safe limit. The results demonstrate that glycoalkaloid‐free mutants can be generated and grown with moderate yield reductions under both greenhouse and field conditions. This suggests a potential for sustainable production of high‐value products, e.g., food‐grade protein and fibre, from starch production side‐streams of SGA knockout tubers.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145311346","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}
Lipid droplets (LDs) serve as the primary storage site for neutral lipids in plant cells, with growing evidence supporting many additional biological roles, such as in lipid homeostasis, signalling, trafficking, inflammatory responses and inter‐organelle communication. While the biogenesis and structure of LDs in seeds and other plant tissues have been well‐documented, the full range of their functions has yet to be elucidated. Plant LDs encapsulate a hydrophobic neutral lipid core, enveloped by a phospholipid monolayer embedded with specific proteins. Despite their tissue‐specific diversity, a range of methods for LD isolation from plant materials has been established, facilitating lipidomic and proteomic characterisation. This knowledge has facilitated studies into the potential applications of LDs, particularly in pharmaceutical biotechnology. This review explores the multifunctional nature and biogenesis of plant LDs, highlights recent advances in LD fractioning from plant materials, explores factors affecting their stability, and discusses the potential of mimicking natural LDs using artificial lipid nano‐droplets (ALNDs) and similar synthetic lipid‐based formulations. It also underscores the significance of LD‐based delivery systems in pharmaceutical applications, emphasising their emerging potential in enhancing drug solubility, bioavailability and targeted delivery. Finally, future research directions are highlighted, focusing on scaling up LD isolation, optimising ALND and other formulations, and investigating their pharmacokinetics and long‐term stability for more widespread clinical applications.
{"title":"Plant Lipid Droplets and Derived Lipidic Nano‐Assemblies: Structure, Biogenesis and Pharmaceutical Applications","authors":"Abdulsamie Hanano, Amal Yousfan, Denis J. Murphy","doi":"10.1111/pbi.70411","DOIUrl":"https://doi.org/10.1111/pbi.70411","url":null,"abstract":"Lipid droplets (LDs) serve as the primary storage site for neutral lipids in plant cells, with growing evidence supporting many additional biological roles, such as in lipid homeostasis, signalling, trafficking, inflammatory responses and inter‐organelle communication. While the biogenesis and structure of LDs in seeds and other plant tissues have been well‐documented, the full range of their functions has yet to be elucidated. Plant LDs encapsulate a hydrophobic neutral lipid core, enveloped by a phospholipid monolayer embedded with specific proteins. Despite their tissue‐specific diversity, a range of methods for LD isolation from plant materials has been established, facilitating lipidomic and proteomic characterisation. This knowledge has facilitated studies into the potential applications of LDs, particularly in pharmaceutical biotechnology. This review explores the multifunctional nature and biogenesis of plant LDs, highlights recent advances in LD fractioning from plant materials, explores factors affecting their stability, and discusses the potential of mimicking natural LDs using artificial lipid nano‐droplets (ALNDs) and similar synthetic lipid‐based formulations. It also underscores the significance of LD‐based delivery systems in pharmaceutical applications, emphasising their emerging potential in enhancing drug solubility, bioavailability and targeted delivery. Finally, future research directions are highlighted, focusing on scaling up LD isolation, optimising ALND and other formulations, and investigating their pharmacokinetics and long‐term stability for more widespread clinical applications.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"35 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145311309","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}
Phytopathogenic fungi secrete a great number of effector proteins into various organelles of host plants and suppress plant immunity through different mechanisms. In this study, we identify SCRE7 as a unique nuclear effector that is essential for U. virens infection. SCRE7 interacts with a transcription factor OsLBD11/12 in rice nuclei. Interestingly, defence responses and rice resistance against bacterial and fungal diseases are negatively modulated by SCRE7, but positively regulated by OsLBD11/12. Consistently, transcriptome analyses revealed that SCRE7 and OsLBD11/12 oppositely regulate multiple immune-related pathways in rice. Furthermore, we demonstrated that OsLBD11/12 activates the transcription of OsCPS2 encoding a positive immune regulator through specifically binding to the cis-element in the OsCPS2 promoter. SCRE7 attenuates the binding of OsLBD11/12 to the cis-element and thereby inhibits the transcriptional activity of the transcription factor and rice immunity. The findings not only represent an unidentified infection strategy in U. virens, but also provide candidate gene targets for the creation of disease-resistant germplasms in rice.
{"title":"An Ustilaginoidea virens Nuclear Effector SCRE7 Inhibits Rice Immunity via Suppressing OsLBD11/12-Promoted Transcription of OsCPS2.","authors":"Chunquan Jiang,Xiaoai Li,Anfei Fang,Yu Fu,Caifeng Li,Haojie Dong,Siwen Yu,Shaoqi Zhang,Dayong Li,Nan Nan,Juan Zhang,Reuben J Peters,Dan Zhao,Wenxian Sun","doi":"10.1111/pbi.70413","DOIUrl":"https://doi.org/10.1111/pbi.70413","url":null,"abstract":"Phytopathogenic fungi secrete a great number of effector proteins into various organelles of host plants and suppress plant immunity through different mechanisms. In this study, we identify SCRE7 as a unique nuclear effector that is essential for U. virens infection. SCRE7 interacts with a transcription factor OsLBD11/12 in rice nuclei. Interestingly, defence responses and rice resistance against bacterial and fungal diseases are negatively modulated by SCRE7, but positively regulated by OsLBD11/12. Consistently, transcriptome analyses revealed that SCRE7 and OsLBD11/12 oppositely regulate multiple immune-related pathways in rice. Furthermore, we demonstrated that OsLBD11/12 activates the transcription of OsCPS2 encoding a positive immune regulator through specifically binding to the cis-element in the OsCPS2 promoter. SCRE7 attenuates the binding of OsLBD11/12 to the cis-element and thereby inhibits the transcriptional activity of the transcription factor and rice immunity. The findings not only represent an unidentified infection strategy in U. virens, but also provide candidate gene targets for the creation of disease-resistant germplasms in rice.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"90 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145305712","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}
Drought priming is a critical agronomic strategy for enhancing plant drought tolerance, yet the optimal priming intensity and transcriptional regulatory mechanisms underlying subsequent drought responses in the tea plant (Camellia sinensis) remain poorly characterised. In this study, we systematically evaluated tea plants exposed to recurrent drought stress under varying priming intensities. Results demonstrated that moderate drought priming specifically conferred superior drought tolerance compared to non-primed controls. Integrated metabolomic and transcriptomic profiling identified flavonoid biosynthesis as the key pathway associated with priming-induced drought resilience. Exogenous flavonoid application and overexpression of six biosynthesis genes (CsCHS, CsCHI, CsFLS, CsDFR, CsANS and CsANR) functionally validated flavonoids' role in drought adaptation. Notably, transcriptional regulators CsYABBY1 and CsMYB114 were identified as hub transcription factors demonstrating transcriptional activation potential towards flavonoid biosynthesis. Combinatorial transient overexpression and silencing assays revealed that both CsYABBY1 and CsMYB114 coordinately upregulate flavonoid biosynthesis genes, redirecting metabolic flux towards flavonoid accumulation to enhance drought tolerance. Multimodal validation through yeast one-hybrid assays, dual-luciferase reporter systems and electrophoretic mobility shift assays, as well as molecular docking, confirmed or simulated direct binding of CsYABBY1 and CsMYB114 to promoter regions of flavonoid biosynthesis genes for transcriptional activation. These findings establish a synergistic regulatory model where CsYABBY1 and CsMYB114 cooperatively enhance flavonoid accumulation through transcriptional reprogramming, thereby conferring acquired drought tolerance. This study provides mechanistic insights for developing adaptive cultivation practices and advances molecular breeding strategies for drought-resilient tea cultivars.
{"title":"CsYABBY1 and CsMYB114 Enhance Acquired Drought Tolerance by Mediating Flavonoid Biosynthesis in Camellia sinensis.","authors":"Caiyun Tian,Chengzhe Zhou,Shengjing Wen,Cheng Zhang,Anru Zheng,Lele Jiang,Niannian Yang,Zhuo Tang,Xiaowen Hu,Zhendong Zhang,Jiaxin Fang,Zhong Wang,Yuqiong Guo","doi":"10.1111/pbi.70399","DOIUrl":"https://doi.org/10.1111/pbi.70399","url":null,"abstract":"Drought priming is a critical agronomic strategy for enhancing plant drought tolerance, yet the optimal priming intensity and transcriptional regulatory mechanisms underlying subsequent drought responses in the tea plant (Camellia sinensis) remain poorly characterised. In this study, we systematically evaluated tea plants exposed to recurrent drought stress under varying priming intensities. Results demonstrated that moderate drought priming specifically conferred superior drought tolerance compared to non-primed controls. Integrated metabolomic and transcriptomic profiling identified flavonoid biosynthesis as the key pathway associated with priming-induced drought resilience. Exogenous flavonoid application and overexpression of six biosynthesis genes (CsCHS, CsCHI, CsFLS, CsDFR, CsANS and CsANR) functionally validated flavonoids' role in drought adaptation. Notably, transcriptional regulators CsYABBY1 and CsMYB114 were identified as hub transcription factors demonstrating transcriptional activation potential towards flavonoid biosynthesis. Combinatorial transient overexpression and silencing assays revealed that both CsYABBY1 and CsMYB114 coordinately upregulate flavonoid biosynthesis genes, redirecting metabolic flux towards flavonoid accumulation to enhance drought tolerance. Multimodal validation through yeast one-hybrid assays, dual-luciferase reporter systems and electrophoretic mobility shift assays, as well as molecular docking, confirmed or simulated direct binding of CsYABBY1 and CsMYB114 to promoter regions of flavonoid biosynthesis genes for transcriptional activation. These findings establish a synergistic regulatory model where CsYABBY1 and CsMYB114 cooperatively enhance flavonoid accumulation through transcriptional reprogramming, thereby conferring acquired drought tolerance. This study provides mechanistic insights for developing adaptive cultivation practices and advances molecular breeding strategies for drought-resilient tea cultivars.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145305710","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}
Heat stress (HS) has become an increasing threat to wheat productivity under global warming. However, the genetic loci for thermotolerance and the underlying molecular mechanisms remain largely unknown. In this study, genetic mapping identified a thermotolerance locus, QMpe.cau‐2D, encoding fatty acid desaturase 8 (FAD8), with the transposable element (TE) insertions present in the promoter region in the thermotolerant cultivar. The expression of TaFAD8‐D was negatively associated with thermotolerance. Loss‐of‐function mutations in TaFAD8 enhanced photosynthetic efficiency, seedling survival rate, and thousand‐grain weight under HS. Transcriptome, fatty acid, and lipid profiling analyses showed that TaFAD8 mutation affected the expression of genes involved in lipid biosynthesis and metabolism to mediate the fatty acid composition and lipid remodelling, thereby maintaining chloroplast membrane fluidity and integrity under HS. TaWRKY71 negatively regulated the transcription of TaFAD8 by binding to its promoter, and mutation of TaWRKY71 reduced photosynthetic efficiency under HS. Our findings identify a beneficial TaFAD8‐D haplotype, uncover its molecular mechanism and regulatory pathways in heat response, and provide a strategy for breeding climate‐resilient wheat varieties.
{"title":"Natural Variation in TaFAD8‐D Promoter Enhances Thermotolerance in Wheat Through Fatty Acid and Lipid Remodelling","authors":"Hongjian Yu, Tianyu Lan, Weiwei Mao, Yongfa Wang, Xiaoyu Zhang, Mengsi Ma, Shuo Chen, Guang Chen, Qiang Li, Zhaorong Hu, Mingming Xin, Yingyin Yao, Weilong Guo, Zhongfu Ni, Qixin Sun, Huiru Peng","doi":"10.1111/pbi.70397","DOIUrl":"https://doi.org/10.1111/pbi.70397","url":null,"abstract":"Heat stress (HS) has become an increasing threat to wheat productivity under global warming. However, the genetic loci for thermotolerance and the underlying molecular mechanisms remain largely unknown. In this study, genetic mapping identified a thermotolerance locus, <jats:italic>QMpe.cau‐2D</jats:italic>, encoding fatty acid desaturase 8 (FAD8), with the transposable element (TE) insertions present in the promoter region in the thermotolerant cultivar. The expression of <jats:italic>TaFAD8‐D</jats:italic> was negatively associated with thermotolerance. Loss‐of‐function mutations in <jats:italic>TaFAD8</jats:italic> enhanced photosynthetic efficiency, seedling survival rate, and thousand‐grain weight under HS. Transcriptome, fatty acid, and lipid profiling analyses showed that <jats:italic>TaFAD8</jats:italic> mutation affected the expression of genes involved in lipid biosynthesis and metabolism to mediate the fatty acid composition and lipid remodelling, thereby maintaining chloroplast membrane fluidity and integrity under HS. TaWRKY71 negatively regulated the transcription of <jats:italic>TaFAD8</jats:italic> by binding to its promoter, and mutation of <jats:italic>TaWRKY71</jats:italic> reduced photosynthetic efficiency under HS. Our findings identify a beneficial <jats:italic>TaFAD8‐D</jats:italic> haplotype, uncover its molecular mechanism and regulatory pathways in heat response, and provide a strategy for breeding climate‐resilient wheat varieties.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"4 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145282786","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}
Ariel H. Tomassi, María Juárez‐Molina, Adriana E. Cisneros, Ana Alarcia, Francesca Orlando, Sara Toledano‐Franco, Silvia Presa, Antonio Granell, Alberto Carbonell
RNA interference (RNAi) is a highly conserved gene silencing mechanism regulating gene expression at transcriptional and post‐transcriptional levels in plants. Synthetic trans‐acting small interfering RNAs (syn‐tasiRNAs) have emerged as powerful tools for highly specific and efficient gene silencing. However, their application in crops has been constrained by the need for transgene integration and the relatively long length of TAS‐derived precursors. Here, we developed a novel syn‐tasiRNA platform for Solanum lycopersicum (tomato) based on minimal precursors targeted by endogenous SlmiR482b microRNA. These minimal precursors, comprising only a 22‐nt miRNA target site, an 11‐nt spacer, and the syn‐tasiRNA sequence(s), effectively produced functional syn‐tasiRNAs in both transgenic and transient virus‐induced gene silencing (syn‐tasiR‐VIGS) systems. To facilitate their broader application, we engineered a series of vectors for high‐throughput cloning and efficient syn‐tasiRNA expression from SlmiR482b‐based minimal precursors in tomato. Our results show that minimal precursors induce robust gene silencing of endogenous tomato genes and confer antiviral resistance to the economically important tomato spotted wilt virus. Furthermore, we show that syn‐tasiR‐VIGS can be applied in a transgene‐free manner through crude extract delivery, leading to efficient silencing of endogenous genes. This study establishes minimal syn‐tasiRNA precursors as a versatile and efficient tool for precision RNAi in tomato, with applications in functional genomics and crop improvement.
{"title":"Precision RNAi in Tomato Using Synthetic Trans‐Acting Small Interfering RNAs Derived From Minimal Precursors","authors":"Ariel H. Tomassi, María Juárez‐Molina, Adriana E. Cisneros, Ana Alarcia, Francesca Orlando, Sara Toledano‐Franco, Silvia Presa, Antonio Granell, Alberto Carbonell","doi":"10.1111/pbi.70410","DOIUrl":"https://doi.org/10.1111/pbi.70410","url":null,"abstract":"RNA interference (RNAi) is a highly conserved gene silencing mechanism regulating gene expression at transcriptional and post‐transcriptional levels in plants. Synthetic trans‐acting small interfering RNAs (syn‐tasiRNAs) have emerged as powerful tools for highly specific and efficient gene silencing. However, their application in crops has been constrained by the need for transgene integration and the relatively long length of <jats:italic>TAS</jats:italic>‐derived precursors. Here, we developed a novel syn‐tasiRNA platform for <jats:styled-content style=\"fixed-case\"><jats:italic>Solanum lycopersicum</jats:italic></jats:styled-content> (tomato) based on minimal precursors targeted by endogenous SlmiR482b microRNA. These minimal precursors, comprising only a 22‐nt miRNA target site, an 11‐nt spacer, and the syn‐tasiRNA sequence(s), effectively produced functional syn‐tasiRNAs in both transgenic and transient virus‐induced gene silencing (syn‐tasiR‐VIGS) systems. To facilitate their broader application, we engineered a series of vectors for high‐throughput cloning and efficient syn‐tasiRNA expression from SlmiR482b‐based minimal precursors in tomato. Our results show that minimal precursors induce robust gene silencing of endogenous tomato genes and confer antiviral resistance to the economically important tomato spotted wilt virus. Furthermore, we show that syn‐tasiR‐VIGS can be applied in a transgene‐free manner through crude extract delivery, leading to efficient silencing of endogenous genes. This study establishes minimal syn‐tasiRNA precursors as a versatile and efficient tool for precision RNAi in tomato, with applications in functional genomics and crop improvement.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145282785","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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