Plant Direct最新文献

{"title":"A Reference-Free Algorithm Discovers Regulation in the Plant Transcriptome.","authors":"Elisabeth Meyer, Evan V Saldivar, Marek Kokot, Bo Xue, Sebastian Deorowicz, Seung Y Rhee, Julia Salzman","doi":"10.1002/pld3.70061","DOIUrl":"10.1002/pld3.70061","url":null,"abstract":"<p><p>Most plant genomes and their (post-)transcriptional regulation remain unknown. We used SPLASH-a new, reference genome-free sequence variation detection algorithm-to analyze transcriptional and post-transcriptional regulation from RNA-seq data. We discovered allelic variation in expression during maize pollen development and imbibition-dependent cryptic splicing in Arabidopsis seeds. SPLASH enables discovery of novel regulatory mechanisms, including differential regulation of genes from parental haplotypes of hybrids, without the use of alignment to a reference genome.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 4","pages":"e70061"},"PeriodicalIF":2.3,"publicationDate":"2026-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13066497/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147675956","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional Divergence of the Arg/N-Degron Pathway Between the Crop <i>Brassica rapa</i> and the Model Plant <i>Arabidopsis thaliana</i>.","authors":"Brian C Mooney, Pablo Garcia, Shreenivas Kumar Singh, Emmanuelle Graciet","doi":"10.1002/pld3.70158","DOIUrl":"https://doi.org/10.1002/pld3.70158","url":null,"abstract":"&lt;p&gt;&lt;p&gt;The ubiquitin-dependent Arg/N-degron pathway relates the stability of a substrate protein to the nature of its N-terminal amino acid residue or its biochemical modifications, with some N-terminal residues being recognized by specific E3 ubiquitin ligases, resulting in the ubiquitylation and degradation of the substrate protein. Work in the model plant &lt;i&gt;Arabidopsis thaliana&lt;/i&gt; has shown that the Arg/N-degron pathway is a key regulator of plant responses to hypoxia, which can be either physiological or a stress in the context of waterlogging or submergence. The role of the Arg/N-degron pathway in hypoxia response is mediated via the oxygen-dependent degradation of group VII ETHYLENE RESPONSE FACTOR (ERFVII) transcription factors, which act as the master regulators of the hypoxia response program in plants. Analysis of Arabidopsis mutants for different enzymatic components of the Arg/N-degron pathway has also revealed its roles in the regulation of responses to other abiotic stresses (e.g., salt stress), as well as to pathogens. Although much has been learned from studies in Arabidopsis about the functions of the Arg/N-degron pathway, very little is known about this pathway in crops, including in Brassica crops such as oilseed rape, cabbage, or turnip. To determine functional similarities and divergence of the Arg/N-degron pathway between Arabidopsis and Brassica crops, we isolated and characterized the first Arg/N-degron pathway mutants in &lt;i&gt;Brassica rapa&lt;/i&gt; (turnip, pak choi), a diploid Brassica crop closely related to oilseed rape. We focused on two enzymatic components, namely, the arginine-transferases (&lt;i&gt;ATE&lt;/i&gt;s) and the E3 ubiquitin ligase &lt;i&gt;PROTEOLYSIS6&lt;/i&gt; (&lt;i&gt;PRT6&lt;/i&gt;). Our results show both similarities and divergence of function for these Arg/N-degron pathway components in &lt;i&gt;B. rapa&lt;/i&gt; compared to Arabidopsis. Specifically, &lt;i&gt;ATE&lt;/i&gt; mutants in &lt;i&gt;B. rapa&lt;/i&gt; arrest their development at the seedling stage, which contrasts with the mild phenotypic defects of the equivalent Arabidopsis mutants. Double mutant lines for two of the three &lt;i&gt;PRT6&lt;/i&gt; genes in &lt;i&gt;B. rapa&lt;/i&gt; indicated a constitutive activation of hypoxia response genes at the transcriptional level, as shown in the single &lt;i&gt;prt6&lt;/i&gt; mutant in Arabidopsis. However, contrary to Arabidopsis, the &lt;i&gt;B. rapa&lt;/i&gt; double mutants were more sensitive to waterlogging and hypoxia and did not show differential response to salt stress or to biotic stress compared to the wild type. The functional divergence identified likely reflects variability in each species in the substrate repertoire and/or in the regulation of pathways or targets downstream of Arg/N-degron pathway substrates. Such differences could be driven by direct selective pressures at N-termini (e.g., gain or loss of a destabilizing N-terminal residue) or by species-specific proteases that may generate destabilizing neo-N-termini after cleavage. These similarities and differences highlight the difficulties in tra","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 3","pages":"e70158"},"PeriodicalIF":2.3,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13103730/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147779073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systems analysis of leaf anatomy and transcriptome using unsupervised machine learning provides insight on photosynthesis development and evolution in non-model grasses.","authors":"Santiago Prochetto, Georgina Stegmayer, Anthony J Studer, Renata Reinheimer","doi":"10.1002/pld3.70022","DOIUrl":"https://doi.org/10.1002/pld3.70022","url":null,"abstract":"<p><p>The C₄ photosynthesis includes intriguing leaf anatomies. The current model supports the placement of C₃-C₄ intermediates as a middle point in the evolutionary trajectory from C₃ to C₄ photosynthesis. The known determinants involved in the differentiation of divergent photosynthetic leaves arose from the comparative analysis between both ends, C₃ and C₄ species. However, much more could be known if evolutionarily close species were analyzed together with intermediate species using advanced-omic approaches. In the present work, by combining leaf anatomical traits and transcriptomic data with machine learning methods, we provided insights on gene regulatory networks involved in complex leaf anatomical characteristics in non-model grasses of subtribe Otachyriinae. For that, self-organizing maps (SOMs) were developed to group genes and phenotypic traits into clusters (neurons) according to their behavior along the leaf developmental gradient. The analysis allowed us to identify a set of genes as potential enablers of key anatomical trait differentiation related to bundle sheath (BS) cell size, vein density, and the interface between mesophyll and BS cells. At the same time, we identified genes that displaced together with the adjustment of the BS cell area suggesting a possible role in the evolution of this distinctive leaf anatomical trait.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 3","pages":"e70022"},"PeriodicalIF":2.3,"publicationDate":"2026-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12976145/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147444697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Untargeted Metabolomics Reveals Organ-Specific and Extraction-Dependent Metabolite Profiles in Endemic Tajik Species <i>Ferula violacea</i> Korovin.","authors":"Sylhiya Mavlonazarova, Kenneth Acosta, Rinat Abzalimov, Saidbeg Satorov, Vyacheslav Dushenkov","doi":"10.1002/pld3.70123","DOIUrl":"10.1002/pld3.70123","url":null,"abstract":"<p><p><i>Ferula violacea</i> Korovin, an endemic Tajikistani plant with purported medicinal properties, remains understudied. This study employs untargeted metabolomics to characterize the metabolite profiles of ethanol extracts and juices from <i>F. violacea</i> roots and seeds. In total, 540 distinct metabolites are putatively identified, 419 of which are previously unreported in the <i>Ferula</i> genus, representing a substantial expansion of its known chemical diversity. The most abundant metabolites are terpenoids, amino acid derivatives, and alkaloids. A particularly abundant group of daucane sesquiterpenoids, sharing a common (6-methyl-azulen-4-yl)cyclohexanecarboxylate substructure, is identified, including known metabolites such as ferutidin and ferutinin. Comparative analysis reveals organ-specific metabolic specialization: roots are enriched in terpenoids, whereas seeds exhibit higher concentrations of alkaloids and amino acids. Additionally, processing methods influence metabolite composition, with ethanol extracts being rich in terpenoids and amino acids, and juices displaying a greater diversity of phenylpropanoid-derived compounds. These findings expand the phytochemical richness of <i>F. violacea</i> and suggest its potential as a valuable source of bioactive compounds for pharmacological exploration.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 3","pages":"e70123"},"PeriodicalIF":2.3,"publicationDate":"2026-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12962795/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147378313","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Low-Cost Custom-Built Flow Meters for Plant Hydraulic Conductance: Validation of Accuracy, Precision, and Reproducibility.","authors":"Jessica Samson-Tshimbalanga, Catherine Périé, Morgane Urli","doi":"10.1002/pld3.70154","DOIUrl":"https://doi.org/10.1002/pld3.70154","url":null,"abstract":"<p><p>Measurement of xylem hydraulic conductance provides access to xylem hydraulic conductivity and vulnerability to cavitation, two key traits for assessing plant sensitivity to environmental stressors. We evaluated the performance of custom-built low-cost pressure drop flow meters through nearly 1200 measurements across devices, laboratories, reservoir heights (10, 25 and 45 cm, used to induce pressure head and drive water flux) and PEEK tubing of hydraulic contrasting resistances. Flow meters were interchangeable, with mean differences generally < 3.5% and never exceeding 5%, with 88.9% of comparative tests showing no significant difference. Under recommended conditions (25-45 cm pressure head, downstream-to-upstream pressure ratio ≈0.5), precision reached 1%-7% coefficient of variation. Accuracy, assessed against reference values obtained by water displacement, was also strong, with 68% of measurements deviating by < 5% from reference values and over 78% when measured at {greater than or equal to}25 cm. At 10 cm, performance declined because sensor deviations represented a larger fraction of pressure differential, and low-resistance PEEK tubing increased absolute but not relative error. Validated flow meters proved portable, affordable (≈2500 CAD), and reliable. Their low cost, open-source interface, and publicly available construction protocol make them accessible to laboratories with limited resources, enabling reproducible multi-laboratory studies of plant hydraulics and fostering international collaborations.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 2","pages":"e70154"},"PeriodicalIF":2.3,"publicationDate":"2026-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12928992/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147309430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptomic and Metabolic Insight Into Flavonoid Biosynthesis Underlying Black and Yellow Seed Coat Color Variation in Soybean (<i>Glycine max</i>).","authors":"Kahee Moon, Prakash Basnet, Seung Young Choi, Beom-Soon Choi, Grace Lachica, Nam-Il Park, Kyong-Cheul Park, Taeyoung Um, Ik-Young Choi","doi":"10.1002/pld3.70153","DOIUrl":"https://doi.org/10.1002/pld3.70153","url":null,"abstract":"<p><p>Soybean (<i>Glycine max</i>) seed coat color variation is determined by the accumulation of flavonoid-derived pigments, although the molecular mechanisms underlying this trait remain poorly understood. This study integrated RNA sequencing (RNA-Seq) and high-performance liquid chromatography (HPLC)-based metabolite measurements to investigate black and yellow seed coat soybean lines derived from the same genetic background. Metabolite analysis revealed significantly higher total phenolic content (TPC), total flavonoid content (TFC), total anthocyanin content (TAC), total proanthocyanidin content (TPAC), and antioxidant activity (DPPH, ABTS) in black seed coats, whereas yellow seed coats exhibited elevated total isoflavone content (TIC). RNA-seq at 110 days after sowing (DAS) identified differential expression of flavonoid pathway genes associated with these metabolic differences. Genes upregulated in black seed coats included flavanone 3-hydroxylase (<i>F3H</i>), anthocyanidin synthase (<i>ANS</i>), UDP-glycosyltransferases (<i>UGT78D2</i>, <i>UGT79B6</i>), and glutathione <i>S</i>-transferase (<i>GSTF11</i>), encoding enzymes reported to function in anthocyanin biosynthesis, glycosylation, and vacuolar transport, respectively. Conversely, leucoanthocyanidin reductase (<i>LAR</i>) genes showed higher expression in yellow seed coats despite lower proanthocyanidin (PA) levels, whereas <i>LAC5</i> exhibited black seed-specific expression consistent with potential PA polymerization activity. R2R3-MYB transcription factor genes along with small heat shock protein genes (<i>sHSPs</i>) were also upregulated in black seed coats, suggesting candidate regulatory roles in pigmentation and stress responses. Cytochrome P450 genes showed preferential expression in yellow seed coats, consistent with isoflavonoid pathway activation. Together, these findings elucidate the genetic and metabolic regulation of seed coat color in soybean and identify candidate genes relevant for functional breeding and genomics research.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 2","pages":"e70153"},"PeriodicalIF":2.3,"publicationDate":"2026-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921420/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147271796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome-Wide Analysis of the lncRNAs and miRNAs Involved in Flower Development in Radish.","authors":"LingJun Wu, Xiaobo Luo, Yadong Li, Yueyue Jin, Wanping Zhang","doi":"10.1002/pld3.70155","DOIUrl":"https://doi.org/10.1002/pld3.70155","url":null,"abstract":"<p><p>Radish (<i>Raphanus sativus</i> L.) is an important root vegetable in the cruciferous family. The yield and quality of radish is seriously affected by the premature bolting and flowering. Although the microRNAs (miRNAs) in regulating flower development have been established in radish, the identification and characterization of long noncoding RNAs (lncRNAs) have yet to be explored. In this study, miRNAs and lncRNAs in vegetative and flower stage were conducted by RNA-seq and small RNA sequencing, respectively. A total of 5315 differentially expressed genes (DGEs), 263 DElncRNAs, and 38 DEmiRNAs were detected in two stages. GO analysis found that many flower DGEs associated with reproductive process, response to hormone, and pollination were enriched. In total, 202 DElncRNAs and 257 DElncRNAs were found to have potential cis- and trans-regulatory effects on 572 DEmRNAs and 3902 DGEs, respectively. A total of 93 and 82 DEGs were predicted as putative targets of 31 DEmiRNAs and 29 DEmiRNAs, respectively. Five mRNA-lncRNA-miRNA regulatory pairs involved in flowering time regulation were proposed, including miRNA156a-5p, miRNA399b, miRNA novel-23, miRNA164c-5p, and miRNA165a-5p. The qRT-PCR results showed that four mRNAs, three lncRNAs, and three miRNAs were consistent with the results of RNA-seq and small RNA sequencing. Transient overexpression of miR156a-5p significantly inhibited the expression levels of <i>RsSPL10</i>, <i>RsSPL15</i>, lncRNAs RsLinc1162, and RsLinc214. The results showed that miR156 co-expressed with <i>RsSPL10</i> and <i>RsSPL15</i> significantly inhibited the luciferase activity of <i>RsSPL10</i> and <i>RsSPL15</i> genes, indicating miR156 can directly target <i>RsSPL10</i> and <i>RsSPL15</i> and inhibit their expression. These findings provide a theoretical foundation for further elucidating the molecular regulation mechanism of mRNAs, lncRNAs, and miRNAs in bolting and flowering in radish.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 2","pages":"e70155"},"PeriodicalIF":2.3,"publicationDate":"2026-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12916260/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147271810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional Interaction Between Toc75 POTRA1 Domain and Tic22-III in the Intermembrane Space During Chloroplast Protein Import.","authors":"Rajneesh Singhal, Danny J Schnell","doi":"10.1002/pld3.70138","DOIUrl":"10.1002/pld3.70138","url":null,"abstract":"<p><p>Chaperones are essential for facilitating the import of nuclear-encoded precursor proteins into chloroplasts. In the intermembrane space (IMS) of the chloroplasts, this process is mediated by the transport-associated domains (POTRA) of the translocon at the outer envelope membrane (Toc75) and translocon at the inner envelope membrane (Tic22) proteins. The present work aims to understand the interaction between the Toc75 POTRA domain and Tic22 in the IMS and determine their relationship in facilitating protein import. Expression of the POTRA1 domain deleted <i>TOC75</i> (<i>TOC75ΔP1</i>) in the <i>tic22-III</i> mutant background resulted in a more severe phenotype than the individual mutants, indicating that the two proteins functionally interact in the IMS. Using an insulin aggregation assay, we have demonstrated that Tic22-III also possesses chaperone-like activity. In vitro import experiments suggest that <i>TOC75ΔP1/tic22-III</i> plants are compromised in importing stromal and thylakoid membrane proteins. Therefore, we propose that the Toc75 POTRA domains and Tic22-III both provide chaperone activity necessary to prevent the misfolding of incoming pre-proteins, acting as chaperones and facilitating the protein import process through the IMS of the chloroplast.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 2","pages":"e70138"},"PeriodicalIF":2.3,"publicationDate":"2026-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12895209/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146202314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combined Analysis of Transcriptome and Metabolome Explains the Differences in Pulp Color in Jackfruit.","authors":"Jianjun Liang, Xiangwei Ma, Chenxin Yi, Hailan Zhou, Zhuangmin Wei, Xiuguan Tang, Weiyan Ye, Hailing Tang, Pengjin Zhu","doi":"10.1002/pld3.70127","DOIUrl":"10.1002/pld3.70127","url":null,"abstract":"<p><p>The pulp color of jackfruit reflects variations in its nutritional composition and influences market preference. We investigated the mechanism underlying pulp coloration through an integrated transcriptomic and metabolomic analysis of three jackfruit cultivars-light yellow \"THA\", yellow \"GTM\", and orange \"YNH\". Twenty-five differentially accumulated flavonoids and 32 differentially accumulated carotenoids were identified. Naringenin chalcone, eriodictyol, taxifolin, zeaxanthin, and lutein were identified as the key flavonoids and carotenoids associated with the light-yellow tone of THA pulp. Phlorizin and lutein were associated with the yellow tone of GTM pulp, whereas apigenin, luteolin, zeaxanthin, and violaxanthin dipalmitate were the key pigments regulating the orange tone of YNH pulp. Differentially expressed genes involved in flavonoid and carotenoid biosynthesis included <i>PAL</i>, <i>C4H</i>, <i>4CL</i>, <i>F3H</i>, <i>FLS</i>, <i>ANS</i>, <i>ANR</i>, <i>PSY</i>, <i>PDS</i>, <i>ZDS</i>, <i>LCYE</i>, <i>LUT5</i>, <i>ZEP</i>, and <i>VDE</i>. This study provides a foundation for elucidating the molecular mechanisms underlying jackfruit pulp coloration at transcriptional and metabolic levels.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 2","pages":"e70127"},"PeriodicalIF":2.3,"publicationDate":"2026-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12894421/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146202311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting dCas9-SunTag to a Susceptibility Gene Promoter Is Sufficient for CRISPR Interference.","authors":"Zuh-Jyh Daniel Lin, Gabriela L Hernandez, Myia K Stanton, Xingguo Zheng, Kerrigan B Gilbert, Kira M Veley, Greg Jensen, Marisa Yoder, Suhua Feng, Basudev Ghoshal, Jason Gardiner, Ming Wang, Steven E Jacobsen, James C Carrington, Rebecca S Bart","doi":"10.1002/pld3.70128","DOIUrl":"10.1002/pld3.70128","url":null,"abstract":"<p><p>Cassava production in sub-Saharan Africa is severely impacted by diseases. Most pathogens require interaction with host susceptibility factors to complete their life cycles and cause disease. Targeted DNA methylation is an epigenetic strategy to alter gene expression in plants, and we previously reported that a zinc-finger fused to DMS3 could establish methylation at the promoter of <i>MeSWEET10a</i>, a bacterial susceptibility gene, and this resulted in decreased disease. Here, we attempt a similar strategy for cassava brown streak disease. This disease is caused by the ipomoviruses CBSV and UCBSV. These viruses belong to the family <i>Potyviridae</i>, which has been shown extensively to require host eIF4E-family proteins to infect plants and cause disease. We previously found that cassava plants with simultaneous knockout mutations in two <i>eIF4E</i> genes, <i>nCBP-1 and nCBP-2</i>, resulted in decreased susceptibility to CBSD. Here, we report successful simultaneous targeting of both promoters with methylation using a dCas9-DRMcd-SunTag system. However, in contrast to our previous work with <i>MeSWEET10a,</i> controls indicate that CRISPR interference is occurring in these lines and is sufficient for the reduction of gene expression. Future research will use genetic crosses to segregate away the DNA methylation reagents and, if DNA methylation proves heritable, assess whether methylation alone is sufficient to increase resistance to CBSD.</p>","PeriodicalId":20230,"journal":{"name":"Plant Direct","volume":"10 2","pages":"e70128"},"PeriodicalIF":2.3,"publicationDate":"2026-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12877719/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146143306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}