Plant Biotechnology Journal最新文献

{"title":"Sweet Potato Gene Clusters Control Anthocyanin Biosynthesis and Leaf Morphology","authors":"Dong An,&nbsp;Tiechen Shen,&nbsp;Shiyu Wu,&nbsp;Yanshen Li,&nbsp;Weijuan Fan,&nbsp;Mengxiao Yan,&nbsp;Yinghui Meng,&nbsp;Xinyi Wang,&nbsp;Ximing Xu,&nbsp;Zunfu Lv,&nbsp;Ling Yuan,&nbsp;Jun Yang,&nbsp;Guoquan Lu,&nbsp;Hongxia Wang","doi":"10.1111/pbi.70636","DOIUrl":"10.1111/pbi.70636","url":null,"abstract":"<p>Sweet potato (<i>Ipomoea batatas</i>) exhibits diversity in pigmentation and leaf morphology, yet the genetic architecture and regulatory organisation underlying these traits remain poorly resolved, particularly with respect to organ-specific control. We hypothesised that phenotypic variation is governed by clustered genetic modules comprising regulatory and structural genes operating in an organ-specific manner. To test this, we conducted genome-wide association studies (GWAS) using 4.6 million SNPs across 260 diverse accessions, integrated with transcriptomic, haplotype and functional analyses. GWAS identified two tandem clusters of MYB transcription factors on chromosome 5 as the primary regulators of leaf anthocyanin accumulation. Expression profiling, heterologous expression and transcriptional activation assays demonstrated that <i>IbMYB2</i> and <i>IbMYB3</i> function as key transcriptional activators and form a mutually reinforcing regulatory module. In contrast, pigmentation in storage roots was associated with a spatially distinct genomic region enriched in anthocyanin biosynthetic genes, including <i>IbAOMT</i>, <i>Ib3GGT</i> and <i>IbLDOX</i>, indicating different regulations between aerial and underground organs. Comparative genomic analysis further revealed expansion and conservation of MYB clusters in sweet potato, suggesting evolutionary selection for enhanced transcriptional control. In addition, GWAS uncovered a major locus on chromosome 7 controlling leaf shape variation. Functional analyses demonstrated that conserved developmental regulators, including <i>BEL1-like</i> (<i>g29974</i>), <i>WD40</i> (<i>g26165</i>) and <i>LMI1-like</i> (<i>g29859</i>) genes, play causal roles in leaf margin development. CRISPR/Cas9-mediated knockout of <i>g26165</i> directly reduced leaf lobing, confirming its functional importance. These findings reveal clustered regulatory and structural gene modules underlying key agronomic traits and provide insights into the genetic and evolutionary mechanisms driving phenotypic diversification in sweet potato.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4500-4527"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70636","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147506840","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":"Multisite Field Evaluation of Oil Accumulation and Agronomic Performance in Grain and Sweet Sorghums Engineered for Lipid Hyperaccumulation","authors":"Yunzhu Chen,&nbsp;Kiyoul Park,&nbsp;Chunhwa Jang,&nbsp;Jung Woo Lee,&nbsp;Mengyuan Wang,&nbsp;Hyojin Kim,&nbsp;Truyen Quach,&nbsp;Ming Guo,&nbsp;Balasaheb V. Sonawane,&nbsp;Sanbon Ch. Gosa,&nbsp;Thomas Elmo Clemente,&nbsp;Andrew D. B. Leakey,&nbsp;Edgar B. Cahoon,&nbsp;DoKyoung Lee","doi":"10.1111/pbi.70654","DOIUrl":"10.1111/pbi.70654","url":null,"abstract":"<p>Oil sorghum (OS) has been developed by engineering grain (TX430) and sweet (Ramada) genetic backgrounds to accumulate triacylglycerols (TAG) in vegetative tissues as an energy-dense feedstock for sustainable aviation fuel (SAF) and other biofuels. This study evaluated two TX430 OS lines (TxHO-2, TxHO-3) and two Ramada OS lines (RmHO-1, RmHO-2) alongside wild-type (WT) lines in NE and IL over 2 years (2023–2024) to quantify genotype × environment effects on agronomic performance and TAG accumulation. Across four environments, TX430 OS lines showed average TAG concentrations of 15.0 g kg⁻¹ in leaves and 12.8 g kg⁻¹ in stems, approximately 19-fold higher than WT. Ramada OS lines accumulated 26.1 g kg⁻¹ in leaves and 12.3 g kg⁻¹ in stems, approximately 25-fold and 13-fold increases over WT, respectively. OS lines in TX430 exhibited an 18% reduction in biomass (8.4 vs. 9.9 Mg ha⁻¹ for WT), while Ramada OS lines had similar WT biomass (18.3 vs. 19.9 Mg ha⁻¹ for WT). Among TX430 OS lines, TxHO-2 achieved the highest TAG yield (190 kg ha⁻¹), while RmHO-1 led the Ramada lines (335 kg ha⁻¹) due to higher biomass and similar TAG concentration. Enhanced TAG accumulation increased N, P, and K removal in TX430 lines but not in Ramada lines. Structural carbohydrate and ash concentration were unaffected. Overall, results confirm vegetative lipid accumulation as a viable strategy for high-biomass sorghum, supporting its potential as a dual-purpose feedstock for SAF. Future work should focus on minimizing biomass yield penalties and improving nutrient use efficiency in oil sorghum systems.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4546-4560"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70654","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147506837","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":"StALKBH10B-Mediated RNA m6A Modification Inhibits Potato Salt Tolerance by Targeting Flavonoids and ABA Signalling Pathways","authors":"Xuanming Dong,&nbsp;Jianyu Ma,&nbsp;Chenxin Sui,&nbsp;Yan Li,&nbsp;Vadim Khassanov,&nbsp;Hongju Jian,&nbsp;Dianqiu Lv","doi":"10.1111/pbi.70642","DOIUrl":"10.1111/pbi.70642","url":null,"abstract":"<p>N⁶-methyladenosine (m⁶A) is the most prevalent methylation modification present in mRNAs, which has been confirmed to participate in many developmental and biological processes. However, the biological function and specific regulatory mechanism of m⁶A modification in relation to salt tolerance of potato remain obscure. Here, we generated a transcriptome-wide m⁶A map using salt-resistant and salt-sensitive potato varieties under salt stress conditions to uncover patterns of m⁶A methylation in the potato response to salt stress. MeRIP-seq revealed that m⁶A is significantly enriched in the CDS region in potato, by recognising the conserved motifs including RRACH and URRUAY. Numerous differential m⁶A-deposited transcripts have been identified, which were significantly enriched in ABA-signalling and flavonoids biosynthesis pathway in two potato varieties after salt stress. Notably, a positive correlation was observed between the m⁶A enrichment and mRNA abundance based on combined analysis of MeRIP-seq and mRNA-seq. <i>StALKBH10B</i> was identified as an m⁶A demethylase for decreasing m⁶A modification levels, inhibiting mRNA stability and translation efficiency of ABA signal-related genes (<i>StABF3, StAAO3</i>, and <i>StZEP7</i>) and flavonoids biosynthesis genes (<i>StPAL3</i>, <i>StCHS</i>, and <i>StFLS</i>), and overexpression of <i>StALKBH10B</i> suppressed salt resistance in potato. Collectively, we uncover a novel mechanism of post-transcriptional modification involved in affecting salt stress response in potato, via StALKBH10B-mediated m⁶A demethylation on targeted transcripts in the ABA signalling and flavonoids biosynthesis pathway, thereby providing candidate genes for the breeding of stress-tolerant potato cultivars.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4435-4453"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70642","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147478754","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":"Structural Elucidation and Engineering of the (S)-scoulerine 2-O-Methyltransferase Enabling Regioselective Epiberberine Biosynthesis in Coptis chinensis","authors":"Jun Song,&nbsp;Di Liu,&nbsp;Shi Chen,&nbsp;Yufeng Tan,&nbsp;Siyu Wang,&nbsp;Shihan Li,&nbsp;Yutong Qin,&nbsp;Guofeng Li,&nbsp;Xufang Tian,&nbsp;Lan Yang,&nbsp;Chong Yuan,&nbsp;Junbo Gou,&nbsp;Wei Huang,&nbsp;Shilin Chen,&nbsp;Yifei Liu","doi":"10.1111/pbi.70650","DOIUrl":"10.1111/pbi.70650","url":null,"abstract":"<p>Protoberberine alkaloids are a characteristic group of natural products in <i>Coptis</i> plants known for their notable pharmacological activities. However, the structural similarity and the substrate promiscuity of their biosynthetic enzymes have left the precise synthetic pathways remain unclarified, posing challenges to regulate product formation. In this study, we identified CcOMT8, a key enzyme responsible for C2-methoxylation in the biosynthesis of epiberberine in <i>C. chinensis</i>, through methyl jasmonate elicitation analysis and comparative genomics-based microsynteny analysis. Functional characterisation demonstrated that CcOMT8 specifically catalyses 2-<i>O</i>-methylation of (<i>S</i>)-scoulerine, as verified by heterologous expression in both microbial and plant systems. Its lack of activity toward (<i>S</i>)-cheilanthifoline further confirmed the specific route for epiberberine biosynthesis. Structural investigations of CcOMT8 and its complexes revealed key aspects of substrate recognition and a catalytic mechanism mediated by the His253-Asp254-Glu312 triad. Comparative structural analysis with 9-<i>O</i>-methyltransferases indicated that hydrophilic residues and reduced steric hindrance in the substrate binding pocket govern the regioselectivity of CcOMT8. Using focused rational iterative site-specific mutagenesis (FRISM), we developed an optimised mutant, S109L/C250A/L300A, with 4.88-fold enhanced catalytic efficiency. This study elucidates the biosynthetic pathway of epiberberine in <i>Coptis</i>, clarifies the molecular basis of enzyme-directed metabolic flux, and provides efficient biocatalysts for the synthetic biosynthesis of protoberberine alkaloids.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4528-4545"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70650","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147506850","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":"Two Aquaporins Mitigate Growth-Defence Trade-Offs by Facilitating CO2 and H2O2 Transport in Wheat","authors":"Kai Lu,&nbsp;Shuo Qi,&nbsp;Hui Qian,&nbsp;Xiaohan Fu,&nbsp;Ziyang An,&nbsp;Xiaochen Chen,&nbsp;Liyuan Zhang,&nbsp;Shenshen Zou,&nbsp;Lei Chen,&nbsp;Hansong Dong","doi":"10.1111/pbi.70648","DOIUrl":"10.1111/pbi.70648","url":null,"abstract":"<p>The growth-defence trade-offs pose a major challenge to breeding high-yield and disease-resistant crops. Aquaporins are membrane channels that facilitate the transport of water and other small compounds, therefore regulating the growth-defence trade-offs. However, the molecular mechanism that governs the function of aquaporins in the trade-offs remains unclear. Here, we report that <i>Triticum aestivum</i> TaPIP1;6 and TaPIP2;10, aquaporins of the plasma membrane intrinsic protein (PIP) family, function as dual substrate channels that concurrently enhance plant growth and resistance to powdery mildew and English grain aphid. In wheat plants growing under normal conditions, TaPIP1;6 and TaPIP2;10 facilitate CO₂ transport from the atmosphere into wheat cells and promote photosynthesis, which leads to growth enhancement and grain yield increase. In wheat plants under attack by powdery mildew pathogen or English grain aphid, TaPIP1;6 and TaPIP2;10 function as concurrent H₂O₂ transport channels, mediating the influx of apoplastic H₂O₂ into the cytoplasm. In turn, the transported H₂O₂ activates innate immunity, including the MAPK cascade, callose deposition and defence gene expression, and thereby enhances wheat resistance to powdery mildew and the English grain aphid. In essence, co-overexpression of <i>TaPIP1;6</i> and <i>TaPIP2;10</i> exhibits synergistic effects on CO₂ and H₂O₂ transports, further amplifying both yield and resistance traits. Taken together, our results suggest that TaPIP1;6 and TaPIP2;10 function as dual-substrate transporting channels to promote growth by regulating CO₂ transport and to enhance resistance against pathogens and insects via H₂O₂-mediated immune responses. This finding provides crucial genetic targets for breeding crop varieties combining high yield with resistance traits.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4487-4499"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70648","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147502299","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":"Redundant Glycerol-3-Phosphate Acyltransferases Regulate Thermo-Sensitive Genic Male Sterility in Rice","authors":"Siqi Cheng,&nbsp;Hai Zheng,&nbsp;Yunlu Tian,&nbsp;Chaolong Wang,&nbsp;Xiaowen Yu,&nbsp;Kun Dong,&nbsp;Jian Wang,&nbsp;Jiayu Lu,&nbsp;Keyi Chen,&nbsp;Xiaodong He,&nbsp;Kun Shao,&nbsp;Junwen Gao,&nbsp;Bowen Yao,&nbsp;Yang Hu,&nbsp;Zhenwei Xie,&nbsp;Ling He,&nbsp;Zhao Xie,&nbsp;Yuantao Zhu,&nbsp;Song Guo,&nbsp;Dekun Lei,&nbsp;Anqi Jian,&nbsp;Xufei Zhu,&nbsp;Hao Yu,&nbsp;Shihao Zhang,&nbsp;Minrui Chen,&nbsp;Yun Chen,&nbsp;Ling Jiang,&nbsp;Yulong Ren,&nbsp;Xiuping Guo,&nbsp;Shijia Liu,&nbsp;Xi Liu,&nbsp;Shanshan Zhu,&nbsp;Zhigang Zhao,&nbsp;Jianmin Wan","doi":"10.1111/pbi.70649","DOIUrl":"10.1111/pbi.70649","url":null,"abstract":"<p>Thermo-sensitive genic male sterility (TGMS) lines play a pivotal role in two-line hybrid rice breeding. However, the availability of elite TGMS germplasm remains limited, and the molecular mechanisms underlying fertility transition of TGMS lines are still poorly understood. Here, we identified a novel TGMS line, <i>Osgpat6.1</i>, which exhibits temperature-dependent fertility transition, with male sterility under high-temperature (HT) and restored fertility under low-temperature (LT). Cytological observations revealed that the TGMS line exhibits disrupted programmed cell death (PCD) of the tapetum under HT conditions. This disruption leads to delayed tapetum degradation and defective pollen exine formation. Map-based cloning identified a premature termination mutation in glycerol-3-phosphate acyltransferase 6.1 (<i>OsGPAT6.1</i>), which is highly expressed in anthers and essential for lipid biosynthesis. Lipidomic profiling revealed that disruption of <i>OsGPAT6.1</i> perturbed lipid metabolism during pollen development. Genetic studies further revealed that male sterility in <i>Osgpat6.1</i> mutant was restored through functional redundancy with its homologues <i>OsGPAT6.2</i> and <i>OsGPAT6.3</i> under LT. Overexpression of <i>OsGPAT6.2</i> or <i>OsGPAT6.3</i> partially restored the fertility of <i>Osgpat6.1</i> mutant under HT. However, simultaneous disruption of all three homologues abolished fertility restoration even under LT. Notably, introgression of the <i>Osgpat6.1</i> allele into diverse rice cultivars produced stable TGMS lines without compromising hybrid vigor. Collectively, these findings demonstrate that dysfunction of GPAT confers TGMS in rice by disrupting glycerolipid metabolism and provides TGMS germplasm to facilitate two-line hybrid rice breeding.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4689-4703"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70649","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147584074","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":"Beyond Starch: Towards a Scalable Potato Platform for Molecular Farming","authors":"Izabela Anna Chincinska,&nbsp;Dorota Sołtys-Kalina,&nbsp;Audrey Y.-H. Teh","doi":"10.1111/pbi.70616","DOIUrl":"10.1111/pbi.70616","url":null,"abstract":"<p>Thirty-five years after the first recombinant protein was produced in potato and 30 years after clinical trials of edible vaccines from its tubers, the crop is being reconsidered as a molecular farming chassis. Potatoes can accumulate recombinant proteins in tubers, enabling long-term storage and simplified logistics. Clonal propagation, access to minitubers and microtubers, and an established production infrastructure further distinguish the platform. Limited pollen dispersal and reliance on vegetative propagation also provide biosafety advantages. Despite these features, potato lost ground to other hosts due to low expression levels, high downstream processing (DSP) costs in water- and starch-rich tissues and limited scalability relative to seed-propagated crops. We review the technical and economic factors behind this decline and assess new opportunities to overcome them. Recent advances include refined expression cassettes, ER and secretory pathway engineering (including targeted glycoengineering), multigene stacking, genome editing and enzyme-assisted DSP. Seed-propagated diploid potatoes and alternative <i>Solanum</i> germplasm offer additional chassis options. Together with the emergence of start-ups revisiting potato as a production platform, these advances point to practical routes for re-evaluating its role in plant molecular farming. We argue that a rationally engineered, diploid-based ‘bioreactor potato’ could complement existing hosts and re-establish relevance in specific niches of next-generation biomanufacturing.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4619-4639"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70616","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147536459","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":"Single-Cell CRISPR: An Efficient Strategy for Decoding Plant Cis-Regulatory Complexity","authors":"Yuanhang Zhang,&nbsp;Xuanxuan Luo,&nbsp;Hongbin Li,&nbsp;Shuangxia Jin,&nbsp;Xianlong Zhang,&nbsp;Maojun Wang","doi":"10.1111/pbi.70647","DOIUrl":"10.1111/pbi.70647","url":null,"abstract":"<p>The generation of complex traits involves the coordinated interplay of multiple gene networks. Elucidating the function of transcriptional <i>cis</i>-regulatory elements (CREs) in regulating gene expression is crucial for understanding complex regulatory pathways and improving our ability to modify macro-phenotypes. While traditional bulk sequencing approaches rely on tissue or cell population aggregates, single-cell transcriptomics provides a more precise perspective by capturing cell-type-specific information. The integration of single-cell technology with genome-wide genetic screening, particularly through the single-cell CRISPR (scCRISPR) system, enables the identification of critical regulatory elements and provides novel insights into gene-expression control mechanisms. Here, we summarise recent advances in diverse strategies for functional genome analysis using the scCRISPR system, with an emphasis on its potential to revolutionise single-cell genetic screening of CREs. We also explore the challenges and opportunities for applying these approaches in plant research.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4454-4467"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70647","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147483659","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":"Tuning Xylan Polymerisation Enhanced Fibre Digestibility Without Biomass Loss in Sheepgrass (Leymus chinensis)","authors":"Mengjie Zhao,&nbsp;Zhuolin Liu,&nbsp;Jianli Wang,&nbsp;Lei Chen,&nbsp;Shanjie Tang,&nbsp;Zhelong Lin,&nbsp;Tong Li,&nbsp;Fengqi Cheng,&nbsp;Linlin Mu,&nbsp;Dongmei Zhang,&nbsp;Xu Zhuang,&nbsp;Duofeng Pan,&nbsp;Zhongbao Shen,&nbsp;Dengpan Bu,&nbsp;Xianwei Song,&nbsp;Yihua Zhou,&nbsp;Shuaibin Zhang,&nbsp;Baocai Zhang,&nbsp;Xiaofeng Cao","doi":"10.1111/pbi.70651","DOIUrl":"10.1111/pbi.70651","url":null,"abstract":"<p><i>Leymus chinensis</i> (sheepgrass), a dominant perennial grass of the Eurasian Steppe, is a crucial source of carbohydrates and energy for ruminants. However, the lignocellulose recalcitrance severely limits its digestibility. Here, we targeted xylan, a major hemicellulose interacting with cellulose and lignin in cell wall. To improve digestibility, we knocked out two <i>IRREGULAR XYLEM 14-LIKE (LcIRX14L)</i> genes, which encode key enzymes for xylan biosynthesis. The <i>Lcirx14L</i> knockout mutants exhibited reduced xylan molecular weight but produced similar amounts of biomass to wild-type plants. Simon's staining and cellulase binding assays indicated enhanced cellulose accessibility in the <i>Lcirx14L</i> mutant. In addition, the <i>Lcirx14L</i> plant showed increased saccharification efficiency, dry matter and NDF digestibility, and production of volatile fatty acids by <i>in vitro</i> fermentation. This study thus provides novel insight for engineering forage grasses with both high yield and quality by modifying xylan polymerization.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4484-4486"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70651","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147502298","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":"Precise Creation of Elite Multilocular Germplasm Using a CBE NG System in Brassica napus","authors":"Huailin Li,&nbsp;Limin Hu,&nbsp;Yang Yu,&nbsp;Sukanta Bala,&nbsp;Yungu Zhai,&nbsp;Yang Yang,&nbsp;Xiaoxiao Shen,&nbsp;Hanzi He,&nbsp;Chuchuan Fan","doi":"10.1111/pbi.70658","DOIUrl":"10.1111/pbi.70658","url":null,"abstract":"&lt;p&gt;Multilocular silique is a desirable agricultural trait that has great potential in developing high-yield &lt;i&gt;Brassica&lt;/i&gt; crops due to increased seeds per silique and enhanced shatter resistance (Fan et al. &lt;span&gt;2014&lt;/span&gt;). A few multilocular lines of &lt;i&gt;Brassica&lt;/i&gt; crops have been identified in nature, such as the multilocular ‘Yellow Sarson’ in &lt;i&gt;B. rapa&lt;/i&gt; and ‘Duoshi’ in &lt;i&gt;B. juncea&lt;/i&gt; (Liu &lt;span&gt;2000&lt;/span&gt;). The trait is conferred by recessive nuclear genes. In &lt;i&gt;B. rapa&lt;/i&gt;, a causal C-to-T nucleotide substitution in a &lt;i&gt;CLAVATA3&lt;/i&gt; (&lt;i&gt;CLV3&lt;/i&gt;) homologue, causing a Pro9-to-Leu change in the CLE motif, confers the multilocular phenotype (Fan et al. &lt;span&gt;2014&lt;/span&gt;), accompanied by a significant improvement in plant yield (Figure S1). However, no stable natural multilocular germplasm has been identified in the important oilseed species &lt;i&gt;B. napus&lt;/i&gt;, hindering breeding.&lt;/p&gt;&lt;p&gt;Previously, we pioneered the application of CRISPR/Cas9 for generating knockout mutants of &lt;i&gt;BnaCLV3&lt;/i&gt; homologues in &lt;i&gt;B. napus&lt;/i&gt; (Yang et al. &lt;span&gt;2018&lt;/span&gt;). The resulting double homozygous mutation of &lt;i&gt;BnA04.CLV3&lt;/i&gt; and &lt;i&gt;BnC04.CLV3&lt;/i&gt; (&lt;i&gt;a&lt;/i&gt;&lt;sup&gt;&lt;i&gt;n&lt;/i&gt;&lt;/sup&gt;&lt;i&gt;a&lt;/i&gt;&lt;sup&gt;&lt;i&gt;n&lt;/i&gt;&lt;/sup&gt;&lt;i&gt;c&lt;/i&gt;&lt;sup&gt;&lt;i&gt;n&lt;/i&gt;&lt;/sup&gt;&lt;i&gt;c&lt;/i&gt;&lt;sup&gt;&lt;i&gt;n&lt;/i&gt;&lt;/sup&gt;) developed multilocular siliques with more seeds than wild-type (WT). However, these lines exhibited pleiotropic developmental defects (excessive leaves, flattened stems, enlarged inflorescence apices, distorted siliques), limiting breeding utility (Figure 1). The specific amino acid substitution in &lt;i&gt;B. rapa&lt;/i&gt; likely offers a better strategy. Cytidine base editors (CBEs), enabling precise C-to-T conversions without double-strand breaks, provide a promising tool for creating optimised germplasm (Hu et al. &lt;span&gt;2020&lt;/span&gt;; Wu et al. &lt;span&gt;2020&lt;/span&gt;).&lt;/p&gt;&lt;p&gt;To develop high-yielding multilocular rapeseed, this study employed CBE systems to achieve precise single-base substitution from P&lt;sub&gt;CCT&lt;/sub&gt; to L&lt;sub&gt;CTT&lt;/sub&gt; at the ninth amino acid of the BnaCLV3 CLE domain. However, the target site lacked available NGG-PAM. We therefore engineered a CBE-nCas9 system (CBE-Cas9NG) recognising NG-PAM. The base editor (Anc689 APOBEC, nCas9NG, and UGI) from Anc689BE4max-nCas9NG (Wang et al. &lt;span&gt;2019&lt;/span&gt;) replaced the Cas9 in the pYLCRISPR/Cas9P35S-H (Yang et al. &lt;span&gt;2018&lt;/span&gt;), creating pBGE21. A universal sgRNA1 was designed for targeting both &lt;i&gt;BnaCLV3&lt;/i&gt; copies was cloned with AtU6-26-driven sgRNA or enhanced sgRNA (esgRNA) cassettes into pBGE21, yielding pBGE21a/b (Figure 1A).&lt;/p&gt;&lt;p&gt;The vectors were introduced into J9707 via &lt;i&gt;Agrobacterium&lt;/i&gt;-mediated hypocotyl transformation, yielding 163 positive transgenic seedlings (Figure 1B). Average mutation rates of 7.05% for sgRNA and 4.10% for esgRNA indicate that the CBE-Cas9NG system combined with the sgRNA expression cassette achieved satisfactory editing efficiency (Figure 1B). Genotyping further confirmed that not only the intended C6 ","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 7","pages":"4716-4718"},"PeriodicalIF":12.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70658","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147598150","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}