Shi Liang, Weiqinlan Wang, Chengxin Tan, Lin Zhou, Zhi Ou, Yan Qu
{"title":"Multi-omics analysis reveals the potential role of MbDDC in tissue-specific alkaloid biosynthesis and distribution in Meconopsis betonicifolia","authors":"Shi Liang, Weiqinlan Wang, Chengxin Tan, Lin Zhou, Zhi Ou, Yan Qu","doi":"10.1111/tpj.70515","DOIUrl":"10.1111/tpj.70515","url":null,"abstract":"<div>\u0000 \u0000 <p>Plants of the <i>Meconopsis</i> (<i>Meconopsis</i> spp.), endemic to the Qinghai-Tibet Plateau, are prized in traditional Tibetan medicinal herbs for their bioactive alkaloids, particularly their antispasmodic and analgesic properties. To elucidate the mechanisms underlying tissue-specific alkaloid accumulation in <i>Meconopsis betonicifolia</i>, we integrated metabolomic and transcriptomic analyses across four organs (roots, stems, leaves, and flowers) and functionally characterized the rate-limiting enzyme MbDDC-3. Our results demonstrate that roots are the primary site of alkaloid accumulation, with codeinone and salutaridine identified as key intermediates in the isoquinoline pathway. Eleven differentially expressed genes (DEGs) were strongly correlated with these metabolites. Heterologous overexpression of <i>MbDDC-3</i> in tobacco (<i>Nicotiana tabacum</i>) significantly increased total alkaloid by 274% in roots (<i>P</i> < 0.05), with (S)-cis-<i>N</i>-methylstylopine and its precursors (tyramine/dopamine) significantly enriched. Notably, MbDDC-3 protein contains a non-classical nuclear localization signal (NLS)—RLKPAAIFNRKLG—located near its C-terminal region and exhibits key residue substitutions compared to lowland species, suggesting adaptive evolution under high-altitude stress. Collectively, this study reveals how <i>M. betonicifolia</i> optimizes alkaloid distribution for ecological fitness, while offering a genetic tool for metabolic engineering of medicinal alkaloids.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145231110","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}
Geoffrey Thomson, Benoit Mermaz, Cintia H. D. Sagawa, Chan-Yi Ivy Lin, Megan Tachev, Valentin Joly, Vivian F. Irish, Yannick Jacob
{"title":"Enzymatic depletion of transposable elements in sequencing libraries and its application for genotyping multiplexed CRISPR-edited plants","authors":"Geoffrey Thomson, Benoit Mermaz, Cintia H. D. Sagawa, Chan-Yi Ivy Lin, Megan Tachev, Valentin Joly, Vivian F. Irish, Yannick Jacob","doi":"10.1111/tpj.70501","DOIUrl":"https://doi.org/10.1111/tpj.70501","url":null,"abstract":"<div>\u0000 \u0000 <p>Whole-genome sequencing has become a common strategy to genotype individual plants of interest. Although a limited number of genomic regions usually need to be surveyed with this strategy, excess sequencing information is almost always generated at an appreciable financial cost. Repetitive sequences (e.g., transposons), which can account for more than 80% of the genome of some plants, are often not required in these genotyping projects. Therefore, strategies that enrich DNA coding for the protein-coding genes prior to sequencing can lower the cost to obtain sufficient sequence information. Here, we present the development and application of methylation-sensitive reduced representation sequencing (MsRR-Seq), which relies on the cytosine methylation-sensitive restriction enzyme MspJI to deplete constitutive heterochromatic DNA before library construction. By applying MsRR-Seq to citrus and maize, we show that protein-coding genes can be enriched in sequencing datasets. We then describe the application of MsRR-Seq to facilitate the identification of complex mutants from populations of citrus plants resulting from multiplex CRISPR/Cas9 editing of four genes. Overall, this work demonstrates an easy and low-cost method to enrich non-repetitive DNA in high-throughput sequencing libraries, an approach that is especially useful for large plant genomes with an excessively high proportion of methylated repetitive sequences.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145224380","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}
Jong Ho Kim, Ju Hee Kim, Geun Beom Park, Seo Young Kim, Cheol Seong Jang
{"title":"Rice E3 ligase OsRFPH2-16 acts as a negative regulator to mediate the degradation of OsPIP1;1 under salt stress","authors":"Jong Ho Kim, Ju Hee Kim, Geun Beom Park, Seo Young Kim, Cheol Seong Jang","doi":"10.1111/tpj.70491","DOIUrl":"10.1111/tpj.70491","url":null,"abstract":"<div>\u0000 \u0000 <p>Soil salinity has a significant negative effect on rice productivity. We characterized the <i>Oryza sativa RING Finger Protein H2-type-16 gene</i> (<i>OsRFPH2-16</i>), which plays a negative role in response to salinity. The transcript levels of <i>OsRFPH2-16</i> decreased under saline conditions. OsRFPH2-16 was expressed in the ER and tonoplasts of rice protoplasts. In addition, OsRFPH2-16 exhibited E3 ligase activity in an <i>in vitro</i> ubiquitination assay, whereas the mutant OsRFPH2-16<sup>C188A</sup> E3 ligase did not exhibit any activity. We constructed <i>OsRFPH2-16</i>-overexpressing (OX-2 and OX-4) and CRISPR/Cas9-mediated <i>OsRFPH2-16</i>-knockout (KO-4 and KO-16) plants and evaluated their salt responses. Under salt stress, <i>OsRFPH2-16-</i>knockout plants exhibited improved salt tolerance, characterized by low Na<sup>+</sup> accumulation, high non-antioxidant content, and dynamic changes in the expression levels of Na<sup>+</sup> transporter genes, compared with wild-type and <i>OsRFPH2-16</i>-overexpression plants. The aquaporin OsPIP1;1, an interacting partner, was identified using yeast two-hybridization, bimolecular fluorescence complementation, and pull-down assays. Degradation of OsPIP1;1 by the E3 ligase OsRFPH2-16 via the 26S proteasome system was confirmed through an <i>in vitro</i> degradation assay with the inhibitor MG132. These findings support that the E3 ligase functions as a negative regulator, leading to reduced Na<sup>+</sup> accumulation in salt stress responses.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145228322","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}
Ignacio Cota, Silvia Moschin, Elisabetta Offer, Irene Martínez-Fernández, Francesco Magnanimi, Barbara Ambrose, Sebastiano Nigris, Barbara Baldan, Cristina Ferrándiz, Soraya Pelaz
{"title":"NGATHA carpel development genes evolved in the common ancestor of seed plants","authors":"Ignacio Cota, Silvia Moschin, Elisabetta Offer, Irene Martínez-Fernández, Francesco Magnanimi, Barbara Ambrose, Sebastiano Nigris, Barbara Baldan, Cristina Ferrándiz, Soraya Pelaz","doi":"10.1111/tpj.70488","DOIUrl":"https://doi.org/10.1111/tpj.70488","url":null,"abstract":"<div>\u0000 \u0000 <p>The evolution of the carpel, the defining feature of angiosperms, remains a fundamental question in plant biology. Understanding how this organ originated is crucial because it underpins the reproductive success and diversity of flowering plants. Here, we investigated the functional conservation between gymnosperms and angiosperms of key transcription factors involved in carpel development. We found that <i>Ginkgo biloba</i> homologs can functionally substitute for their angiosperm counterparts in stigma development. We discovered that <i>GbRAV5</i> is related to angiosperm <i>NGA</i> genes, challenging previous notions that these are exclusive to angiosperms, and we found a parallel loss of the AP2 domain in gymnosperms providing a rare snapshot of how protein families evolve. Conserved protein interactions and overlapping expression patterns of <i>GbRAV5</i> and <i>GbHEC</i> in <i>Ginkgo</i> ovules suggest that the molecular toolkit for carpel development was largely present in the last common ancestor of seed plants, offering new insights into the evolution of reproductive structures.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145224382","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":"GmSPX5 regulates arbuscular mycorrhizal colonization and phosphate acquisition through modifying transcription profile and microbiome in soybean","authors":"Xingqi Yang, Yuanyuan Li, Tianqi Wang, Zipei Li, Qingli Zhuang, Cuiyue Liang, Xiurong Wang, Jiang Tian","doi":"10.1111/tpj.70511","DOIUrl":"10.1111/tpj.70511","url":null,"abstract":"<div>\u0000 \u0000 <p>Symbiosis with arbuscular mycorrhizal (AM) fungi is a crucial strategy for plant adaptation to low phosphorus (P) stress. However, the mechanisms underlying how phosphate (Pi) signaling regulators participate in AM colonization remain largely unknown in soybean (<i>Glycine max</i>). In this study, the expression of <i>GmSPX5</i>, one member of the SPX (SYG1/Pho81/XPR1) family, was induced by AM fungal inoculation in soybean roots. Furthermore, the expression of <i>GmSPX5</i> seems to overlap with AM infection structures through analyzing GUS activity of transgenic soybean plants harboring <i>Pro<sub>GmSPX5</sub>:GUS</i>. Four transgenic lines with <i>GmSPX5</i> overexpression (OX8 and OX12) and suppression (Ri9 and Ri11) were subsequently used to examine the functions of <i>GmSPX5</i> on AM symbiosis and Pi acquisition. Despite no difference between Ri and wild-type (WT), the overexpression of <i>GmSPX5</i> significantly increased AM colonization as reflected by 8.4% in OX8 and 8.7% in OX12, respectively. Consistently, the dry weight and total P content of OX8 and OX12 were higher than WT. Furthermore, a total of 3483 genes were found to exhibit differential expression patterns in roots between OX12 and WT, including genes related to linolenic acid metabolism and flavonoid metabolism. Meanwhile, the composition of the bacterial community in the roots of OX12 was distinct from that in WT through β-diversity analysis. Particularly, an ASV19 (<i>Sphingomonadales</i>) was enriched in OX12 roots, which was positively related to total P content and AM fungi colonization. Taken together, these results highlight that <i>GmSPX5</i> can regulate AM symbiosis, as well as Pi acquisition in soybean. Our findings advance the understanding of SPX functions in plant–microbe interaction.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145228284","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 chromosome-level genome of Stylosanthes guianensis provides insights into genome evolution and environmental adaptation","authors":"Liangliang He, Zhihao Wu, Cuiling Liu, Jianhui Chen, Jiahao Zheng, Jieyi Li, Chao Liu, Chen Liang, Hanying Li, Ci Ren, Yihua Wang, Shu Chen","doi":"10.1111/tpj.70505","DOIUrl":"https://doi.org/10.1111/tpj.70505","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Stylosanthes guianensis</i> is a leguminous forage crop of significant economic importance, primarily distributed in tropical and subtropical regions. It exhibits strong adaptability to various stresses, yet the genetic basis underlying this trait remains unclear. In this study, we constructed the first chromosome-scale reference genome of <i>S. guianensis</i> using a combination of Nanopore and Hi-C sequencing technologies. The assembled genome size is 1254 Mb, with 10 pseudochromosomes. Using Nanopore full-length transcriptome data, we generated high-quality transcript-level gene annotations, identifying 36 585 gene models and 110 601 transcripts. The repetitive sequences in <i>S. guianensis</i> account for 79.16% of the genome, with the extensive expansion of <i>Gypsy</i> elements in long terminal repeats contributing to its genome size enlargement. Comparative genomic and transcriptomic analyses revealed that flavonoid metabolism plays a pivotal role in stress adaptation, providing new insights into the genetic basis of stress tolerance. Additionally, we generated whole-genome methylation profiles under cold treatment and control conditions, offering valuable data for future epigenomic research. These findings provide essential molecular resources for understanding stress resilience in <i>S. guianensis</i> and advancing its molecular breeding.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145224381","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":"Deubiquitinase ZmUBP5 is essential for maize kernel development","authors":"Haixiao Dong, Li Zhang, Hao Chen, Yuan Jiang, Pingping Wang, Chaoyue Wang, Shengzhong Su, Xiaohui Shan, Shipeng Li, Hongkui Liu, Zecheng Zuo, Yaping Yuan","doi":"10.1111/tpj.70482","DOIUrl":"https://doi.org/10.1111/tpj.70482","url":null,"abstract":"<div>\u0000 \u0000 <p>Ubiquitination and deubiquitination are critical post-translational modifications in eukaryotes, playing essential roles in various aspects of plant growth and development. However, their involvement in maize kernel development remains largely unexplored. In this study, we characterized a novel maize kernel mutant (<i>ubp5-ems#1</i>) induced by ethyl methanesulfonate, which exhibited a smaller size, arrested embryo, smaller endosperm, and pigment deficiencies. Through bulked segregant analysis, we identified a stop-gained mutation in the <i>ZmUBP5</i> gene as the cause of these phenotypic abnormalities. This was further validated by allelic confirmation and CRISPR/Cas9 knockout. ZmUBP5, identified as a ubiquitin-specific protease with deubiquitinase (DUB) activity, was localized to the nucleus, and the stop-gained mutation disrupted these functions. Protein interaction and degradation assays revealed that ZmUBP5 interacts with and may deubiquitinate ZmEMB140. The stop-gained mutations in <i>ZmEMB140</i> resulted in smaller kernels with reduced embryo and endosperm size, as well as lethal seedlings. ZmEMB140 is likely a spliceosome-associated factor, interacting with six other proteins involved in pre-mRNA processing. Overall, this study underscores the critical roles of the DUB ZmUBP5 and its potential substrate ZmEMB140 in maize kernel development.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145224272","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":"BnaC04.bZIP16 can be phosphorylated and inhibited by BnaA06.SnRK2 and negatively regulates the accumulation of fatty acids in Brassica napus","authors":"Wei Hui, Shuangshuang Li, Jinhao Ding, Qianru Li, Yuhong Chen, Yanhui Wang, Xupeng Guo, Chengming Fan, Zanmin Hu","doi":"10.1111/tpj.70506","DOIUrl":"https://doi.org/10.1111/tpj.70506","url":null,"abstract":"<p><i>Brassica napus</i> is an important oil crop with a significant amount of fatty acid in its seeds. However, the molecular mechanisms of fatty acid synthesis and accumulation are still poorly understood. In this study, we identified a transcription factor, <i>BnaC04.bZIP16</i>, which belongs to the G subfamily of the bZIP family, that negatively regulates the accumulation of fatty acid in <i>B. napus</i>. BnaC04.bZIP16 directly binds to the promoters of <i>BnaA08.OLEOSIN1</i> and <i>BnaC03.PDH-E1 BETA</i> to regulate seed fatty acid content. Furthermore, BnaC04.bZIP16 can be phosphorylated by the BnaA06.SnRK2. BnaA06.SnRK2-dependent phosphorylation can weaken BnaC04.bZIP16 protein stability as well as the DNA-binding ability. Interestingly, overexpression of <i>BnaA06.SnRK2</i> can significantly promote the accumulation of fatty acid. Our study identified a new function of <i>BnaC04.bZIP16</i> in regulating the accumulation of fatty acids in <i>B. napus</i> and added it to the G subfamily of the bZIP family. Furthermore, our study proposed a regulatory pathway, ‘BnaA06.SnRK2-BnaC04.bZIP16-<i>BnaA08.OLEOSIN1</i>/<i>BnaC03.PDH-E1 BETA</i>’, which provides novel insight into fatty acid biosynthesis.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 1","pages":""},"PeriodicalIF":5.7,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.70506","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145224273","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}
Patrick R Hickland, Katrin Geisler, Shelby Newsad, Marcel Llavero Pasquina, A Caroline Faessler, Gonzalo I Mendoza-Ochoa, Andre Holzer, Payam Mehrshahi, Alison G Smith
{"title":"A new tool for engineering Phaeodactylum tricornutum: the METE promoter drives both high expression and B<sub>12</sub>-tuneable regulation of transgenes.","authors":"Patrick R Hickland, Katrin Geisler, Shelby Newsad, Marcel Llavero Pasquina, A Caroline Faessler, Gonzalo I Mendoza-Ochoa, Andre Holzer, Payam Mehrshahi, Alison G Smith","doi":"10.1111/tpj.70210","DOIUrl":"10.1111/tpj.70210","url":null,"abstract":"<p><p>For advanced metabolic engineering strategies, it is crucial to be able to regulate transgene expression, to prevent potential deleterious effects in the host organism during growth and allow optimisation of production levels. Here, we identified vitamin B<sub>12</sub> (cobalamin)-responsive promoters in the diatom Phaeodactylum tricornutum, a promising biotechnological chassis that readily absorbs this metabolite with minimal physiological impact. Using promoter-reporter constructs, the promoters of the cobalamin acquisition protein 1 (CBA1) and the B<sub>12</sub>-independent form of methionine synthase (METE) were shown to regulate transgene expression in a B<sub>12</sub>-dependent manner. Further characterisation of the METE promoter (P<sub>METE</sub>) demonstrated that it exhibited significantly higher expression levels than several previously characterised promoters, but could be repressed by nanomolar amounts of B<sub>12</sub>, with a dynamic range >100-fold. Tight regulation was demonstrated by the suppression of the lethal ribonuclease, barnase at 1 μg L<sup>-1</sup> B<sub>12</sub>. Reporter expression was doubled when P<sub>METE</sub> was paired with its cognate terminator, compared with the widely used FCPA terminator. Promoter truncations resulted in decreased expression, but no loss of B<sub>12</sub> regulation. A 14 nucleotide motif, present in four copies in P<sub>METE</sub>, was found to be necessary for expression, and when fused to the constitutive FCPA promoter, enhanced expression levels. Transgenic lines expressing the heterologous diterpenoid enzyme, casbene synthase, produced casbene titres of approximately 2 mg L<sup>-1</sup> and this was tuneable by B<sub>12</sub>. This demonstrates the utility of P<sub>METE</sub> in efforts to establish P. tricornutum as an industrial biotechnology production platform.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 2","pages":"e70210"},"PeriodicalIF":5.7,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12538271/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336136","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}
Atheer Balobaid, Wanda M Waterworth, Sophya F Vila Nova, Barry Causier, Vinay Sharma, Madeline R Park, Manish K Pandey, Christopher E West
{"title":"Arabidopsis thaliana FANCONI ANAEMIA I (FANCI) has roles in the repair of interstrand crosslinks and CRISPR-Cas9 induced DNA double strand breaks.","authors":"Atheer Balobaid, Wanda M Waterworth, Sophya F Vila Nova, Barry Causier, Vinay Sharma, Madeline R Park, Manish K Pandey, Christopher E West","doi":"10.1111/tpj.70533","DOIUrl":"10.1111/tpj.70533","url":null,"abstract":"<p><p>DNA repair is crucial for genome stability, in particular for plants which are exposed to high levels of damage arising from UV irradiation, soil pollutants and reactive oxygen species. Damage that affects both strands of the DNA duplex is harder to repair due to both the lack of a template strand and the potential for physical separation of fragmented chromosomes. As such, DNA double-strand breaks (DSBs) and interstrand DNA crosslinks (ICL) are particularly cytotoxic forms of damage. Here we report the functions of FANCONI ANAEMIA I (FANCI), an Arabidopsis thaliana homologue of the mammalian ICL repair protein. We show that in plant cells, as in mammals, FANCI forms a nuclear localised complex with FANCD2. Genetic analysis of plants lacking FANCI displays significant hypersensitivity to the DNA crosslinking reagent mitomycin C. Furthermore, mutation of FANCI in combination with mutations in a second ICL repair factor, METHYL METHANESULFONATE AND UV-SENSITIVE PROTEIN 81 (MUS81), results in increased levels of programmed cell death compared to the corresponding single mutants, revealing roles in maintaining plant genome stability. Sequence analysis of mutational repair of CRISPR-Cas9-induced DSBs revealed that FANCI promotes single nucleotide insertions and reduces longer deletions. This pattern of mutations may reflect roles for FA proteins in replication-coupled repair of a subset of DSBs. Taken together, this analysis finds evidence for multiple roles for FANCI in the maintenance of plant genome stability.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"124 2","pages":"e70533"},"PeriodicalIF":5.7,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12541362/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145342414","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}