The Plant Journal最新文献

{"title":"Identification of UDP-glucosyltransferase involved in the biosynthesis of phloridzin in Gossypium hirsutum","authors":"Xiaomeng Zhang,&nbsp;Xinquan Tian,&nbsp;Junyu Luo,&nbsp;Xiaoyang Wang,&nbsp;Shoupu He,&nbsp;Gaofei Sun,&nbsp;Ruidan Dong,&nbsp;Panhong Dai,&nbsp;Xiao Wang,&nbsp;Zhaoe Pan,&nbsp;Baojun Chen,&nbsp;Daowu Hu,&nbsp;Liru Wang,&nbsp;Baoyin Pang,&nbsp;Aishuang Xing,&nbsp;Guoyong Fu,&nbsp;Baoquan Wang,&nbsp;Jinjie Cui,&nbsp;Lei Ma,&nbsp;Xiongming Du","doi":"10.1111/tpj.17248","DOIUrl":"https://doi.org/10.1111/tpj.17248","url":null,"abstract":"<div>\u0000 \u0000 <p>Phloridzin has various functions, including antioxidant properties and the treatment of diabetes, and has long been used in pharmaceutical and physiological research. The glycosylation of phloretin is a key step in the biosynthesis of phloridzin. In this study, a genome-wide association study (GWAS) based on phloridzin content was applied, and the key gene <i>GhUGT88F3</i> for phloridzin-specific biosynthesis was identified in cotton. A single-base deletion in <i>GhUGT88F3</i> in haplotype I caused a frameshift mutation, leading to premature translation termination and a significant reduction in phloridzin content. Molecular docking revealed important amino acid residues for GhUGT88F3's UDP-glucose transfer activity. Additionally, the transcription factor <i>GhMYB330</i> was found to positively regulate <i>GhUGT88F3</i> expression through population transcriptome analysis and LUC experiment. Moreover, phloridzin content was significantly elevated in both <i>GhUGT88F3</i> and <i>GhMYB330</i> overexpression transgenic plants. This study expands the diversity of UDP-glucosyltransferases in plants and offers a potential strategy for the sustainable production of bioactive compounds with therapeutic potential.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143389078","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":"Defining the heterogeneous composition of Arabidopsis thylakoid membrane","authors":"Andrea Trotta,&nbsp;Sanna Gunell,&nbsp;Azfar Ali Bajwa,&nbsp;Virpi Paakkarinen,&nbsp;Hiroaki Fujii,&nbsp;Eva-Mari Aro","doi":"10.1111/tpj.17259","DOIUrl":"https://doi.org/10.1111/tpj.17259","url":null,"abstract":"<p>Thylakoid membrane (TM) of land plants is organized into an appressed domain (grana), enriched in photosystem (PS) II and a non-appressed domain (stroma lamellae) enriched in PSI. This ultrastructure controls the exciton spillover from PSII to PSI. The bulky machinery required for the biogenesis and repair of TM protein complexes is located in the non-appressed membranes. Thus, the connecting domain (CD) between grana and stroma lamellae is the key player in both the structural and functional integrity of the photosynthetic machinery. In addition, both the grana domain and the stroma lamellae are highly curved at their edges due to the action of the CURVATURE1 (CURT1) proteins, forming a domain distinct from the CD, called the curvature. Here we elucidate the biochemical properties and proteome composition of different thylakoid domains. To this end, the TM of <i>Arabidopsis thaliana</i> (Arabidopsis), isolated both in the natural stacked configuration and in an artificially unstacked configuration to induce a homogeneous protein composition, was solubilized and fractionated, using the mild detergent digitonin (DIG). Using mass spectrometry-based proteomics, we characterize composition, distribution and interaction of proteins involved in TM function in grana, CD and stroma lamellae domains. We find that a subset of thylakoid protein complexes are readily solubilized into small vesicles by DIG and accumulate in a loose pellet (LP) together with CURT1. By combining an extensive biochemical and proteome characterization of the TM fractions we provide an optimized protocol and proteome maps that can be used as a basis for experimental design in photosynthesis research.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17259","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143380779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Arabidopsis thaliana Zn transporter genes ZIP3 and ZIP5 provide the main Zn uptake route and act redundantly to face Zn deficiency","authors":"Valeria Ochoa Tufiño,&nbsp;Maria Almira Casellas,&nbsp;Aron van Duynhoven,&nbsp;Paulina Flis,&nbsp;David E. Salt,&nbsp;Henk Schat,&nbsp;Mark G. M. Aarts","doi":"10.1111/tpj.17251","DOIUrl":"https://doi.org/10.1111/tpj.17251","url":null,"abstract":"<p>In response to Zn deficiency, plants are thought to adjust Zn homeostasis through the coordinated expression of Zn transporters. Zn transporters are identified in the ZIP, HMA and CDF families of cation transporters, although only few are characterized. We determined gene expression over time, root-specific location of expression and phenotypes of single and double loss-of-function mutants of several <i>Arabidopsis thaliana</i> transporters, known to be induced by Zn deficiency. Transcription of Zn transporter genes is induced in the first 6 h of exposure to Zn deficiency. <i>ZIP1</i> is predominantly expressed in the endodermis and stele, <i>ZIP3</i> and <i>ZIP5</i> in the epidermis and cortex, <i>IRT3</i> from epidermis to stele and <i>HMA2</i> in xylem parenchyma. <i>ZIP3</i> and <i>ZIP5</i> act redundantly, with the double-mutant <i>zip3zip5</i> showing high sensitivity to Zn deficiency with low biomass production, expression of other transporter genes, low Zn uptake and increased metal translocation. The root expression map and timing indicate that Zn transporters act complementary in a concerted action to control Zn homeostasis. The lack of strong Zn-deficient phenotypes in single mutants suggests a high level of functional redundancy, best illustrated for <i>ZIP3</i> and <i>ZIP5</i>.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17251","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143380444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rice glycosyltransferase UGT706F1 functions in heat tolerance through glycosylating flavonoids under the regulation of transcription factor MYB61","authors":"Shuman Zhao,&nbsp;Yuqing Ma,&nbsp;Yi Ding,&nbsp;Guangrui Dong,&nbsp;Chonglin Liu,&nbsp;Xinmei Ma,&nbsp;Bingkai Hou","doi":"10.1111/tpj.17252","DOIUrl":"https://doi.org/10.1111/tpj.17252","url":null,"abstract":"<div>\u0000 \u0000 <p>Global metabolic and transcriptional reprogramming is a common event in plant abiotic stress responses, however, the relevant molecular mechanisms remain largely unknown. Here, we characterized the physiological function and molecular mechanism for the rice <i>UGT706F1</i>. We found that <i>UGT706F1</i> can be potently induced by high temperature. Its overexpression can markedly enhance the heat tolerance of rice through improving the capacity of scavenging reactive oxygen species, whereas its functional deletion results in heat sensitivity in rice. To investigate the regulatory mechanism of <i>UGT706F1</i> in response to high temperature, we carried out extensive screening of the in vitro enzymatic activity of UGT706F1 and discovered that UGT706F1 exhibits broad-spectrum activity toward flavonoid compounds. Through targeted flavonoid metabolomics analysis, we further revealed that the overexpression of <i>UGT706F1</i> elevated the content of diverse flavonoids and flavonoid glycosides in rice. Subsequently, via transcriptome analysis, we found that following heat treatment, the overexpression of <i>UGT706F1</i> was capable of enhancing the transcriptional activity of those genes including the flavonoid synthases, heat shock factors, heat shock proteins, glutathione S-transferase, and various antioxidant enzymes. Furthermore, we identified an R2R3 MYB-type transcription factor MYB61 and demonstrated that MYB61 could directly bind the promoter of <i>UGT706F1</i> and activate the transcription of <i>UGT706F1</i>. The overexpression of <i>MYB61</i> also enhanced the heat tolerance and increased flavonoid glycosides. Overall, this study unveiled a novel pathway of the plant heat tolerance response mediated by MYB61-UGT706F1 module and identified a new UGT player for the metabolic and transcriptional regulation under high-temperature circumstance.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143380780","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":"Extensive remodeling during Chlamydomonas reinhardtii zygote maturation leads to highly resistant zygospores","authors":"Martim Cardador,&nbsp;Stephanie Krüger,&nbsp;Susanne Dunker,&nbsp;Alexandra Brakel,&nbsp;Ralf Hoffmann,&nbsp;Raimund Nagel,&nbsp;Torsten Jakob,&nbsp;Reimund Goss,&nbsp;Severin Sasso","doi":"10.1111/tpj.17238","DOIUrl":"https://doi.org/10.1111/tpj.17238","url":null,"abstract":"<p>The unicellular soil alga <i>Chlamydomonas reinhardtii</i> forms diploid zygotes during its sexual cycle. The process of a zygote maturing into a highly resistant zygospore remains poorly understood despite its importance for survival under adverse environmental conditions. Here we describe the detailed timeline of morphological and physiological changes during zygote maturation in darkness on ammonium-free Tris-acetate-phosphate agar plates. The formation of a multilayered cell wall is primarily responsible for the increase in cell size in the first few days after zygote formation. Desiccation and freezing tolerance also develop in the period 3–7 days. Photosynthetic and respiratory activity decrease to reach minimal levels after 7–10 days, accompanied by a partial dedifferentiation of the chloroplast that includes chlorophyll degradation followed by the possible disappearance of the pyrenoid. In contrast to the decreasing concentrations of most carotenoids in the first few days after zygote formation, ketocarotenoids can first be detected after 3 days and their accumulation is completed after 10 days. Furthermore, the zygote degrades a large proportion of its starch and enriches oligosaccharides that may serve as osmoprotectants. The storage lipid triacylglycerol is accumulated at the expense of thylakoid membrane lipids, which mirrors the conversion of a metabolically active cell into a dormant spore on the metabolic level. Taken together, zygote maturation is a multifaceted process that yields mature zygospores after ~ 3 weeks. This work sheds light on the complete time course of the remodeling of a photosynthetically active eukaryotic cell into a dormant, highly resistant spore.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17238","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143379909","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OsPUB9 modulates leaf angle and grain size through the brassinosteroid signaling pathway in rice","authors":"Yonghong Xie,&nbsp;Zhupeng Fan,&nbsp;Xueyan Liang,&nbsp;Kaichong Teng,&nbsp;Zejian Huang,&nbsp;Meiyan Huang,&nbsp;Hong Zhao,&nbsp;Kaizhun Xu,&nbsp;Jianxiong Li","doi":"10.1111/tpj.17230","DOIUrl":"https://doi.org/10.1111/tpj.17230","url":null,"abstract":"<div>\u0000 \u0000 <p>Plant U-box (PUB) E3 ubiquitin ligases are well known for their diverse functions in plant growth and development through physical link to cell signaling hubs to activate regulatory networks. Brassinosteroid (BR) signaling is negatively regulated by a protein kinase GSK2, which interacts with and phosphorylates OsOFP8 (OVATE family protein 8) to regulate BR signaling. Here we identified OsPUB9, a functional E3 ubiquitin ligase, which acts as a negative factor in BR signaling to regulate leaf inclination angle and grain size in rice. OsPUB9 displays autoubiquitination activity and is degraded in response to BR treatment. Interaction with OsUBC13, a rice E2 ubiquitin-conjugating enzyme, suppresses OsPUB9 degradation. OsPUB9 interacts with GSK2, and the interaction reduces autoubiquitination of OsPUB9. Coexpression of OsPUB9 and GSK2 in rice protoplasts suppresses degradation of OsPUB9 but promotes degradation of GSK2. OsPUB9 also interacts with OsOFP8, a positive regulator in BR signaling, and the interaction suppresses degradation of OsPUB9 but facilitates OsOFP8 degradation. Our study reveals that OsPUB9, GSK2, and OsOFP8 form a regulatory network in BR signaling to mediate gene expression, leaf angle, and grain size in rice.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143379910","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":"Chlamydomonas reinhardtii, Volvox carteri and related green algae accumulate ketocarotenoids not in vegetative cells but in zygospores","authors":"Sonja Schwarz,&nbsp;Matthias Bauch,&nbsp;Volker Schmitt,&nbsp;Armin Hallmann,&nbsp;Martin Lohr","doi":"10.1111/tpj.17261","DOIUrl":"https://doi.org/10.1111/tpj.17261","url":null,"abstract":"<p>Zygospores of green alga such as <i>Chlamydomonas reinhardtii</i>, <i>Volvox carteri</i> or <i>Dunaliella salina</i> display a bright orange color indicative of carotenoids, yet there have been no reports on their pigment composition. The genomes of these algae contain genes for homologs of the β-carotene ketolase (BKT) from the well-known astaxanthin producer <i>Haematococcus pluvialis</i>, that were assumed to be pseudogenes, because none of these species has been reported to accumulate astaxanthin or other ketocarotenoids. Here, we show that <i>C. reinhardtii</i> and <i>V. carteri</i> synthesize ketocarotenoids specifically in zygospores. Contrary to the vegetative aplanospores of <i>H. pluvialis</i>, the major ketocarotenoid in zygospores of <i>C. reinhardtii</i> is not astaxanthin but 4-ketolutein. Moreover, the ketocarotenoids in maturing zygospores are not synthesized de novo but from carotenoids of the photosynthetic apparatus liberated by a massive breakdown of thylakoid membranes. In line with this conclusion, incubation of zygospores at 9°C instead of 22°C resulted in a reduced thylakoid breakdown and only low amounts of ketocarotenoids, while the accumulation of storage lipids was less affected. Furthermore, we show the full-length BKT from <i>C. reinhardtii</i> to catalyze the ketolation of both α-carotene and lutein in carotenogenic bacteria. We also detected putative BKT genes in the genomes of various other green algae not yet known to synthesize ketocarotenoids, suggesting a zygospore-specific accumulation of ketocarotenoids to be common among Chlamydomonadales. Our observation that zygospores of <i>C. reinhardtii</i> accumulate ketocarotenoids together with storage lipids sheds light on the physiology of a largely unexplored algal life stage crucial for survival and propagation.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17261","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143379912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SDG8 and HUB2 depositing euchromatin histone marks play important roles in meiosis and crossing-over regulation","authors":"Amit Kumar Singh,&nbsp;K.V.S.K. Arjun Chowdary,&nbsp;Wen-Hui Shen","doi":"10.1111/tpj.17241","DOIUrl":"https://doi.org/10.1111/tpj.17241","url":null,"abstract":"<div>\u0000 \u0000 <p>Histone modifications play critical roles in plant growth and development. Crossing-over (CO) during meiosis, which constitutes a fundamental process ensuring sexual transmission of genetic material to the next generation and, meanwhile, generating diversity within species by creating new chromosome/allele combinations, occurs predominantly in euchromatin, which is enriched in active histone marks such as H3K4me3, H3K36me3, and H2Bub1. In plants, it is known that CO hotspots are correlated with H3K4me3 but the role of H3K36me3 and H2Bub1 during meiosis remains elusive so far. Here, we studied the Arabidopsis (<i>Arabidopsis thaliana</i>) <i>sdg8-1</i> and <i>hub2-2</i> mutants impeded in depositing H3K36me3 and H2Bub1, respectively. Chromosome spreading using 4′,6-diamidino-2-phenylindole (DAPI) staining indicated that male meiotic stages are defective in the <i>sdg8-1</i> mutant, and the defect increases synergistically in the <i>sdg8-1hub2-2</i> double mutant. Defects in meiosis, seed formation, and silique length were also observed by RNAi-knockdown of <i>SDG8</i> using the meiosis-specific gene <i>DMC1</i> promoter. This corroborates to support a <i>bona fide</i> role of active histone marks during meiosis and plant reproduction. Using the tetrad-based visual reporter lines and immunostaining with antibodies against HEI10 and ZYP1, it was found that synapsis and pairing of homologous chromosomes are abnormal and CO rate increases in <i>sdg8</i> mutants, pointing to a repressive role of <i>SDG8</i> in Arabidopsis male meiotic homologous recombination.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143362829","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":"Association analyses reveal both anthropic and environmental selective events during eggplant domestication","authors":"Emmanuel Omondi,&nbsp;Lorenzo Barchi,&nbsp;Luciana Gaccione,&nbsp;Ezio Portis,&nbsp;Laura Toppino,&nbsp;Maria Rosaria Tassone,&nbsp;David Alonso,&nbsp;Jaime Prohens,&nbsp;Giuseppe Leonardo Rotino,&nbsp;Roland Schafleitner,&nbsp;Maarten van Zonneveld,&nbsp;Giovanni Giuliano","doi":"10.1111/tpj.17229","DOIUrl":"https://doi.org/10.1111/tpj.17229","url":null,"abstract":"<p>Eggplant (<i>Solanum melongena</i>) is one of the four most important Solanaceous crops, widely cultivated and consumed in Asia, the Mediterranean basin, and Southeast Europe. We studied the genome-wide association of historical genebank phenotypic data on a genotyped worldwide collection of 3449 eggplant accessions. Overall, 334 significant associations for key agronomic traits were detected. Significant correlations were obtained between different types of phenotypic data, some of which were not obvious, such as between fruit size/yield and fruit color components, suggesting simultaneous anthropic selection for genetically unrelated traits. Anthropic selection of traits like leaf prickles, fruit color, and yield, acted on distinct genomic regions in the two domestication centers (India and Southeast Asia), further confirming the multiple domestication of eggplant. To discriminate anthropic from environmental selection in domestication centers, we conducted a genotype–environment association (GEA) on a subset of georeferenced accessions from the Indian subcontinent. The population structure in this area revealed four genetic clusters, corresponding to a latitudinal gradient, and environmental factors explained 31% of the population structure when the effect of spatial distances was removed. GEA and outlier association identified 305 candidate regions under environmental selection, containing genes for abiotic stress responses, plant development, and flowering transition. Finally, in the Indian domestication center anthropic and environmental selection acted largely independently, and on different genomic regions. These data allow a better understanding of the different effects of environmental and anthropic selection during domestication of a crop, and the different world regions where some traits were initially selected by humans.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17229","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143362623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeted introduction of premature stop codon in plant mitochondrial mRNA by a designer pentatricopeptide repeat protein with C-to-U editing function","authors":"Nikolay Manavski,&nbsp;Eslam Abdel-Salam,&nbsp;Serena Schwenkert,&nbsp;Hans-Henning Kunz,&nbsp;Andreas Brachmann,&nbsp;Dario Leister,&nbsp;Jörg Meurer","doi":"10.1111/tpj.17247","DOIUrl":"https://doi.org/10.1111/tpj.17247","url":null,"abstract":"<p>RNA editing is a crucial post-transcriptional modification in endosymbiotic plant organelles, predominantly involving C-to-U conversions. Pentatricopeptide repeat (PPR) proteins play a key role in this process. To establish a system for gene expression manipulation in genetically inaccessible mitochondria, we engineered a synthetic PPR protein, dPPR-<i>nad7</i>-DYW, to induce <i>de novo</i> C-to-U editing in the <i>NADH dehydrogenase subunit 7</i> (<i>nad7</i>) mRNA of <i>Arabidopsis thaliana</i>, thereby creating a premature stop codon. This designer protein, composed of 13 P-type PPR domains, was fused with the DYW-type cytidine deaminase domain from <i>Physcomitrium patens</i> PpPPR_56 and programmed to bind a specific <i>nad7</i> mRNA segment. <i>In vitro</i> binding assays confirmed the specificity of dPPR-<i>nad7</i>-DYW for its target sequence. When expressed in Arabidopsis plants, dPPR-<i>nad7</i>-DYW achieved up to 85% editing efficiency at the target site, successfully introducing a premature stop codon in <i>nad7</i> mRNA. This resulted in reduced polysome loading of <i>nad7</i> transcripts and a phenotype characteristic of mitochondrial complex I dysfunction. RNA-sequencing revealed potential off-target editing events, albeit at lower frequencies. Our study demonstrates the successful application of an editing factor with a synthetic P-type PPR tract targeting a <i>de novo</i> editing site in plant mitochondria, achieving high editing efficiency. This approach opens new avenues for manipulating organellar gene expression and studying mitochondrial gene function in plants and other eukaryotes.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 3","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17247","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143362827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}