Kim N Janssen,Paul Bolay,Adrian Tüllinghoff,Jörg Toepel,Daniel Spindler,Bruno Bühler,Pia Lindberg
{"title":"Engineering Cyanobacteria for High-Yield Photosynthetic Isoprene Production With Long-Term Phenotypic Stability.","authors":"Kim N Janssen,Paul Bolay,Adrian Tüllinghoff,Jörg Toepel,Daniel Spindler,Bruno Bühler,Pia Lindberg","doi":"10.1111/pbi.70395","DOIUrl":"https://doi.org/10.1111/pbi.70395","url":null,"abstract":"In light of the looming climate crisis, a key cornerstone for a sustainable bioeconomy is photosynthetic production of chemicals and fuels from CO2 and water, powered by sunlight. Isoprene is a five-carbon volatile hydrocarbon with industrial use as a feedstock for rubber production and chemical synthesis and is, at present, generated from crude oil sources. It is, however, possible to produce isoprene using photoautotrophic microorganisms such as cyanobacteria, heterologously expressing the plant enzyme isoprene synthase. Here, we have employed diverse metabolic engineering strategies to develop new strains of the unicellular cyanobacterium Synechocystis sp. PCC 6803 capable of high-level isoprene production from CO2, and have characterised the resulting strains regarding growth, stability and productivity. The new isoprene-producing strains address several challenges in large-scale photobiotechnological production such as genetic and metabolic stability, biosafety and thermotolerance. Moreover, we tested photosynthetic terpenoid production in photobioreactors under process-relevant conditions, achieving the highest volumetric productivities reported so far for a terpene or terpenoid product in cyanobacteria, reaching 148 mg L-1 day-1. Furthermore, we identified and discussed process limitations, laying the foundation for further strain and process engineering towards highly efficient and stable cyanobacterial hydrocarbon production at large scale without selection pressure.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"158 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145246363","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}
Eliza P. I. Loo, José C. Huguet‐Tapia, Michael Selvaraj, Melissa Stiebner, Britta Killing, Marcel Buchholzer, Van Schepler‐Luu, Thomas Hartwig, Sandra P. Valdéz Gutierrez, Madlen I. Rast‐Somssich, Boris Szurek, Joe Tohme, Paul Charraviaga, Frank F. White, Bing Yang, Wolf B. Frommer
{"title":"Removal of Transgenes and Evaluation of Yield Penalties in Genome Edited Bacterial Blight Resistant Rice Varieties","authors":"Eliza P. I. Loo, José C. Huguet‐Tapia, Michael Selvaraj, Melissa Stiebner, Britta Killing, Marcel Buchholzer, Van Schepler‐Luu, Thomas Hartwig, Sandra P. Valdéz Gutierrez, Madlen I. Rast‐Somssich, Boris Szurek, Joe Tohme, Paul Charraviaga, Frank F. White, Bing Yang, Wolf B. Frommer","doi":"10.1111/pbi.70332","DOIUrl":"https://doi.org/10.1111/pbi.70332","url":null,"abstract":"Bacterial blight (BB) of rice, caused by <jats:styled-content style=\"fixed-case\"><jats:italic>Xanthomonas oryzae</jats:italic></jats:styled-content> pv. <jats:italic>oryzae</jats:italic> (Xoo), is one of the major drivers of yield losses in Africa and Asia. Xoo secretes TAL‐effectors (TALe) that induce host SWEET sucrose uniporter by binding to the effector binding element (EBE) of <jats:italic>SWEET</jats:italic> promoters, likely required for Xoo reproduction and virulence. We had multiplex edited the EBEs of three SWEET genes to prevent TALe binding, producing genome‐edited (GE'd) rice mega‐varieties (IR64, Ciherang‐Sub1 for Asia and Komboka for Africa) that were resistant to a wide spectrum of Xoo strains. Here, we report comprehensive analyses of the GE'd lines, including evaluation of agronomic performance in multi‐location multi‐season experimental field plots under different fertilisation regimes and tests for the presence/absence of foreign DNA/transgene in the offspring of GE'd lines (IR64‐BC1T6, Ciherang‐Sub1‐BC1T5, Komboka‐T3). Various strategies were evaluated, including herbicide tolerance, PCR, DNA gel blotting, whole genome sequencing (WGS), and specific tests stipulated by country‐specific biosafety guidelines. Different WGS technologies were evaluated and also used to identify the heritability of the edits, single‐nucleotide polymorphisms (SNPs), and insertions/deletions (indels) that might have resulted from somaclonal variation and potential GE‐induced off‐target mutations. Complete genome reference sequences for the parental lines IR64, Ciherang‐Sub1, and Komboka are provided. In the field experiments, the GE'd lines did not show performance defects. Together, the results indicate that select GE'd lines do not contain foreign DNA or transgene fragments and fulfil the requirements for treatment equivalent to classical breeding lines in countries such as India and Kenya.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"17 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145235381","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}
Marie‐Emilie A. Gauthier, Kylie Shand, Satomi Hayashi, Peter M. Waterhouse, Roberto A. Barrero, Felipe F. de Felippes
{"title":"MicroRNA‐Induced Gene Silencing (MIGS): A Tool for Multi‐Gene Silencing and Targeting Viruses in Plants","authors":"Marie‐Emilie A. Gauthier, Kylie Shand, Satomi Hayashi, Peter M. Waterhouse, Roberto A. Barrero, Felipe F. de Felippes","doi":"10.1111/pbi.70401","DOIUrl":"https://doi.org/10.1111/pbi.70401","url":null,"abstract":"Since its discovery, RNA interference (RNAi, also known as gene silencing) has been a key tool to downregulate gene expression in plants for a range of applications, including protection against viruses. Many of these applications require the silencing of multiple genes concomitantly. Multi‐gene silencing, however, can be difficult to achieve owing to challenges in generating single RNAi constructs targeting unrelated genes or due to molecular instability linked to those constructs. Here, we show that microRNA‐induced gene silencing (MIGS) can overcome many of these limitations and can be an important tool for multi‐gene silencing in plants. We demonstrate how MIGS targeting several genes enhances the RNAi‐based inhibition of one or more viruses. We also define several key features for optimising the use of MIGS in plants, including modular design, effective targeting length and phased first‐base composition.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"107 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145235380","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}
Laming Pei,Yaqing Yang,Zhe Wang,Wencheng Duan,Ning Liu,Zhaohua Ding,Hui Li
{"title":"ZmmiR1432-ZmCML21-ZmPMA2 Module Affects Maize Low Phosphate Tolerance via Regulating Organic Acid Secretion.","authors":"Laming Pei,Yaqing Yang,Zhe Wang,Wencheng Duan,Ning Liu,Zhaohua Ding,Hui Li","doi":"10.1111/pbi.70385","DOIUrl":"https://doi.org/10.1111/pbi.70385","url":null,"abstract":"Phosphorus is indispensable in agricultural production. The growing global demand for food necessitates the development of crops with enhanced phosphorus utilisation efficiency. However, the molecular mechanisms coordinating phosphorus utilisation efficiency in plants remain incompletely characterised. MicroRNAs, pivotal regulators of plant developmental and physiological processes, have emerged as key targets for deciphering the regulatory networks underlying low phosphate (Pi) tolerance. Herein, we delineate the regulatory role of ZmmiR1432 in maize and elucidate its mechanistic basis in conferring low Pi tolerance. Suppression of ZmmiR1432 markedly improved tolerance to Pi deficiency via enhanced organic acid exudation, whereas its overexpression had the opposite effect. It is also indicated that ZmmiR1432 regulates low Pi tolerance through direct modulation of its target gene, ZmCML21, a calmodulin-like protein coding gene that also plays a key role in organic acid secretion and Pi-deficiency response. Metabolomic and transcriptomic analyses revealed that overexpression of ZmCML21 severely affected organic acid secretion and altered the expression of genes involved in the citrate cycle (TCA cycle). Furthermore, it is demonstrated that ZmCML21 directly interacts with plasma membrane H+-ATPase (ZmPMA2). Overexpression of ZmPMA2 phenocopied the ZmmiR1432 knockdown plants and ZmCML21 overexpression plants. Collectively, our findings uncover a ZmmiR1432-ZmCML21 regulatory module that governs low Pi tolerance by modulating ZmPMA2 activity, thereby influencing organic acid secretion and ultimately determining Pi use efficiency. These results provide mechanistic insights and actionable genetic targets for improving Pi use efficiency in maize through molecular breeding and genetic engineering.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145229080","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}
{"title":"The RNA m6A Methyltransferase PvFIP37 Activates Vacuole Transport and Glutathione Detoxification Pathways to Increase Switchgrass Cd Tolerance.","authors":"Mengzhuo Lin,Lei Wang,Bowen Liu,Huayue Liu,Wenwu Qian,Die Zhou,Hui Zang,Binglin Hong,Yunwei Zhang,Jishan Jiang","doi":"10.1111/pbi.70377","DOIUrl":"https://doi.org/10.1111/pbi.70377","url":null,"abstract":"Cadmium (Cd) pollution is a growing global issue that poses significant threats to ecosystems and human health. N6-methyladenosine (m6A) plays an essential role in regulating plant growth and stress responses, but the biological functions and regulatory mechanisms of m6A core enzymes under Cd stress remain largely unexplored. In this study, we demonstrate that PvFIP37 (WTAP human homologue protein) enhances Cd tolerance in switchgrass by activating PvYSL7, PvYSL17, PvABCC4 and PvABCC9 to transport Cd to the vacuoles. Additionally, PvFIP37 stabilises PvGST3 and PvGSTU6 within the glutathione S-transferase detoxification pathway, leading to increased GST activity and a higher GSH/GSSG ratio, thus alleviating oxidative damage in switchgrass under Cd stress. We have further shown that PvMTA interacts with PvFIP37 and targets the same set of genes as PvFIP37 to enhance switchgrass Cd tolerance. In summary, our study illustrates a novel mechanism of m6A methyltransferase-mediated Cd tolerance in switchgrass and provides valuable genetic resources for breeding Cd tolerant plants.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"81 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145235668","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}
Van‐Hung Bui, Joshua C. Wood, Brieanne Vaillancourt, John P. Hamilton, Lemor H. Carlton, Thu‐Thuy T. Dang, C. Robin Buell, Chenxin Li
{"title":"Single Cell Multi‐Omics Reveals Rare Biosynthetic Cell Types in the Medicinal Tree Camptotheca acuminata","authors":"Van‐Hung Bui, Joshua C. Wood, Brieanne Vaillancourt, John P. Hamilton, Lemor H. Carlton, Thu‐Thuy T. Dang, C. Robin Buell, Chenxin Li","doi":"10.1111/pbi.70386","DOIUrl":"https://doi.org/10.1111/pbi.70386","url":null,"abstract":"","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"20 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226597","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}
{"title":"Photoluminescence Properties of Lignin With a Genetically Introduced Luminophore in a Transgenic Hybrid Aspen That Overproduces Feruloyl-CoA 6'-Hydroxylase.","authors":"Masatsugu Takada,Shota Horinouchi,Naning Wang,Mikiko Uesugi,Shinya Kajita","doi":"10.1111/pbi.70390","DOIUrl":"https://doi.org/10.1111/pbi.70390","url":null,"abstract":"Lignin, a major cell-wall component of woody biomass, exhibits photoluminescent (PL) properties. Controlling the intensity and colour of the PL is essential for producing lignin-based value-added materials. Herein, we modify the PL properties of lignin via genetic engineering of novel luminophore structures. Feruloyl-CoA 6'-hydroxylase (F6'H1) is a 2-oxoglutarate-dependent dioxygenase that catalyses the conversion of feruloyl-CoA, an intermediate of the biosynthesis pathway of monolignol, into 6'-hydroxyferuloyl-CoA, the precursor of scopoletin. To modify the lignin PL properties, the F6'H1 gene (F6'H1) from Arabidopsis thaliana is overexpressed in the hybrid aspen (Populus tremula × tremuloides T89), incorporating scopoletin into the lignin molecule. Cellulolytic enzyme lignin (CEL) was isolated from transgenic aspens with different overexpression levels of F6'H1 and evaluated for its PL properties. In N.N-dimethylformamide solution, CEL from the F6'H1-overexpressed aspen emitted clear PL with higher intensity and a longer wavelength than the wild-type CEL. Size exclusion chromatography revealed a wide molar mass distribution of the chromophore. Interestingly, the PL of the CEL from the F6'H1 transgenic lines was limitedly quenched in low polar solvents and at high concentrations. The CEL from F6'H1 emitted obvious PL not only in solution but also in polymer film. Furthermore, the CEL of F6'H1 lines exhibited a reversible photodimerisation reaction characteristic of coumarins. These results suggest that genetic engineering can incorporate new luminophores such as scopoletin into lignin, thus producing value-added materials.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"214 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145209298","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}
Yiming Zhang, Peng Zhu, Long Wu, Zhaohui Li, Yongshun Huang, Mingyu Wang, Kaiying Gu, Peng Wang, Jun Li, Huihua Wan, Hailong Zhang, Jianghua Chen, Ruikun He, Wei Sun, Lixin Li
{"title":"Unveiling the catalytical roles of three glycosyltransferases in triterpenoid saponins in quinoa.","authors":"Yiming Zhang, Peng Zhu, Long Wu, Zhaohui Li, Yongshun Huang, Mingyu Wang, Kaiying Gu, Peng Wang, Jun Li, Huihua Wan, Hailong Zhang, Jianghua Chen, Ruikun He, Wei Sun, Lixin Li","doi":"10.1111/pbi.70214","DOIUrl":"10.1111/pbi.70214","url":null,"abstract":"<p><p>Saponins constitute a class of characteristic compounds within quinoa grains. In this study, metabolomic analysis identified 20 triterpene saponins, the majority of which were exclusively localized within the pericarp. Furthermore, three crucial glycosyltransferases were identified in quinoa. CqCsl2 was characterized as a 3-O-glucuronosyltransferase, CqUGT73FG1 as a 2'-O-glucosyltransferase and CqUGT74BB3 as a C-28 carboxyl glucosyltransferase involved in triterpenoid glycosylation. To elucidate the catalytic mechanisms of CqUGT74BB3, molecular docking simulations and site-directed mutagenesis experiments revealed His13 and Asp104 as critical catalytic residues. Moreover, C-28 carboxyl glycosylation of oleanolic acid was observed to occur only when the C-3 position was modified with either mono-glucuronidation or di-O-glycosylation. This specific modification strategically positions the C-28 carboxyl group in close proximity to the catalytic centre of CqUGT74BB3 (His13-Asp104), enabling the glycosylation reaction. We successfully synthesized key triterpene saponins, exemplified by ginsenoside Ro, in tobacco. These findings have significant implications for future metabolic engineering endeavours aimed at developing quinoa varieties with enhanced nutritional and medicinal attributes.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":"4214-4227"},"PeriodicalIF":10.5,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144504320","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}