Ural Yunusbaev,Gabriela Romero Campos,Rajendran Sathishraj,Evgenii Liakh,John W Raupp,Aleksey V Zimin,Alina Akhunova,Dal-Hoe Koo,Eduard Akhunov
Aegilops geniculata Roth is a tetraploid (MgMgUgUg; 2n = 4x = 28) wild relative of wheat and a valuable source of genetic diversity for improving agronomic traits. We present a high-quality homoeolog-resolved assembly and annotation of the Ae. geniculata genome and use it to study the function, origin and evolution of the Mg and Ug genomes. Comparative genomics revealed that the Ug genome has undergone extensive structural rearrangements (SRAs), which were inherited from its diploid ancestor. Chromosomes 4Ug experienced the most extensive SRAs, including translocations from chromosomes 1, 2, 6 and 7, as well as a pericentric inversion that repositioned the centromere closer to the chromosome terminus. The Mg genome had two large-scale translocations, which likely occurred after polyploidization or in its immediate diploid ancestor. These SRAs resulted in the redistribution of genes among the homoeologous chromosomes, especially affecting the disease resistance genes. Although SRAs altered the expression and H3K4me3 marks of homoeologous genes relative to non-rearranged regions, the overall balance of homoeolog expression and active chromatin remained stable, suggesting selective pressure to maintain gene dosage balance. Population genomic analyses of Ae. geniculata and its diploid ancestors, Ae. comosa (MM) and Ae. umbellulata (UU), suggest that Ae. geniculata originated in Western Anatolia. The genomic resources developed in this study will accelerate trait discovery, gene mapping and the transfer of beneficial alleles from this wild relative into wheat.
{"title":"The Evolution and Origin of Allotetraploid Aegilops geniculata Revealed by the Homoeolog-Resolved Genome Assembly.","authors":"Ural Yunusbaev,Gabriela Romero Campos,Rajendran Sathishraj,Evgenii Liakh,John W Raupp,Aleksey V Zimin,Alina Akhunova,Dal-Hoe Koo,Eduard Akhunov","doi":"10.1111/pbi.70456","DOIUrl":"https://doi.org/10.1111/pbi.70456","url":null,"abstract":"Aegilops geniculata Roth is a tetraploid (MgMgUgUg; 2n = 4x = 28) wild relative of wheat and a valuable source of genetic diversity for improving agronomic traits. We present a high-quality homoeolog-resolved assembly and annotation of the Ae. geniculata genome and use it to study the function, origin and evolution of the Mg and Ug genomes. Comparative genomics revealed that the Ug genome has undergone extensive structural rearrangements (SRAs), which were inherited from its diploid ancestor. Chromosomes 4Ug experienced the most extensive SRAs, including translocations from chromosomes 1, 2, 6 and 7, as well as a pericentric inversion that repositioned the centromere closer to the chromosome terminus. The Mg genome had two large-scale translocations, which likely occurred after polyploidization or in its immediate diploid ancestor. These SRAs resulted in the redistribution of genes among the homoeologous chromosomes, especially affecting the disease resistance genes. Although SRAs altered the expression and H3K4me3 marks of homoeologous genes relative to non-rearranged regions, the overall balance of homoeolog expression and active chromatin remained stable, suggesting selective pressure to maintain gene dosage balance. Population genomic analyses of Ae. geniculata and its diploid ancestors, Ae. comosa (MM) and Ae. umbellulata (UU), suggest that Ae. geniculata originated in Western Anatolia. The genomic resources developed in this study will accelerate trait discovery, gene mapping and the transfer of beneficial alleles from this wild relative into wheat.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"27 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145477619","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}
Solanum tuberosum L. (potato) is a key food crop, with its tubers serving as an important food source worldwide. Tuber development is a tightly regulated process involving the transition of a hooked stolon (a modified stem) to a tuber following the perception of mobile signals within the stolon tip. While genes like FLOWERING LOCUS T homologue StSP6A and the transcription factors (TF) StPOTH1 , StBRC1b and StBEL5 have been implicated in this process, little is known about cell‐type‐specific gene expression and its regulation during tuber initiation. To further our understanding of tuber initiation and development, we generated single nuclei multi‐ome data (gene expression and chromatin accessibility from the same nucleus) from hooked stolons of tetraploid S. tuberosum cv. Atlantic. Nuclei (20079) were assigned to 27 clusters, representing 10 annotated cell types. Differential chromatin and motif enrichment analysis revealed binding sites of TF families known to play a role in cell type development in Arabidopsis that were enriched in analogous cell types in potato stolon tips. By coupling gene co‐expression and information from differential chromatin analysis, we identified novel TFs with putative roles in stolon vasculature development. Co‐accessibility analysis further uncovered putative regulatory enhancers involved in stolon/tuber development. We identified cells that metabolise starch and used gene co‐expression analysis to uncover novel TFs involved in the transition from source to sink. This dataset of cell‐type‐specific gene expression and accessible chromatin from the same nucleus is a powerful resource for discovering genes and regulatory sequences involved in the earliest stages of tuber development.
{"title":"Single Cell Multiomics of Hooked Potato Stolons Reveals Parallels to Shoot Apical Meristems in Arabidopsis","authors":"Dionne Martin, Joshua C. Wood, C. Robin Buell","doi":"10.1111/pbi.70454","DOIUrl":"https://doi.org/10.1111/pbi.70454","url":null,"abstract":"<jats:styled-content style=\"fixed-case\"> <jats:italic>Solanum tuberosum</jats:italic> </jats:styled-content> L. (potato) is a key food crop, with its tubers serving as an important food source worldwide. Tuber development is a tightly regulated process involving the transition of a hooked stolon (a modified stem) to a tuber following the perception of mobile signals within the stolon tip. While genes like FLOWERING LOCUS T homologue <jats:italic>StSP6A</jats:italic> and the transcription factors (TF) <jats:italic>StPOTH1</jats:italic> , <jats:italic>StBRC1b</jats:italic> and <jats:italic>StBEL5</jats:italic> have been implicated in this process, little is known about cell‐type‐specific gene expression and its regulation during tuber initiation. To further our understanding of tuber initiation and development, we generated single nuclei multi‐ome data (gene expression and chromatin accessibility from the same nucleus) from hooked stolons of tetraploid <jats:styled-content style=\"fixed-case\"> <jats:italic>S. tuberosum</jats:italic> </jats:styled-content> cv. Atlantic. Nuclei (20079) were assigned to 27 clusters, representing 10 annotated cell types. Differential chromatin and motif enrichment analysis revealed binding sites of TF families known to play a role in cell type development in <jats:italic>Arabidopsis</jats:italic> that were enriched in analogous cell types in potato stolon tips. By coupling gene co‐expression and information from differential chromatin analysis, we identified novel TFs with putative roles in stolon vasculature development. Co‐accessibility analysis further uncovered putative regulatory enhancers involved in stolon/tuber development. We identified cells that metabolise starch and used gene co‐expression analysis to uncover novel TFs involved in the transition from source to sink. This dataset of cell‐type‐specific gene expression and accessible chromatin from the same nucleus is a powerful resource for discovering genes and regulatory sequences involved in the earliest stages of tuber development.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"43 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145461531","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}
Drought is the major abiotic stress threatening global crop yields, thus identifying potential candidates with promising breeding value has become a central goal of current breeding programmes. Here, we found that miR164b functions as a negative regulator in plant drought tolerance, whose expression is dramatically inhibited under drought stress. Overexpressing MIR164b reduced the drought tolerance, while STTM164 transgenic seedlings showed enhanced drought tolerance in foxtail millet. We further identified that NAC (NAM‐ATAF1/2‐CUC2) transcription factor SiNAC015 was a target of miR164b. The sinac015 mutants showed attenuated drought tolerance, whereas overexpressing mSiNAC015 (miR164b‐resistant version) improved drought tolerance in foxtail millet. Genetic evidence indicated that SiNAC015 could function in the same pathway as miR164b to mediate drought response by directly repressing the expression levels of SitPRX genes, which encoded peroxidase (POD) involved in reactive oxygen species (ROS) scavenging. Additionally, the superior SiNAC015Hap1 possessing higher SiNAC015 expression was found to be associated with enhanced drought tolerance in foxtail millet. Collectively, our study reveals that the miR164b‐ SiNAC015 module mediates drought stress response and provides a valuable genetic resource for drought‐resistant breeding in foxtail millet.
{"title":"The miR164b ‐ SiNAC015 Module Regulates Drought Tolerance by Scavenging Reactive Oxygen Species in Foxtail Millet","authors":"Tong Xiao, Liang Ma, Yifan Zhang, Linlin Zhang, Shixin Song, Xianmin Diao, Jingjuan Yu","doi":"10.1111/pbi.70430","DOIUrl":"https://doi.org/10.1111/pbi.70430","url":null,"abstract":"Drought is the major abiotic stress threatening global crop yields, thus identifying potential candidates with promising breeding value has become a central goal of current breeding programmes. Here, we found that miR164b functions as a negative regulator in plant drought tolerance, whose expression is dramatically inhibited under drought stress. Overexpressing <jats:italic>MIR164b</jats:italic> reduced the drought tolerance, while STTM164 transgenic seedlings showed enhanced drought tolerance in foxtail millet. We further identified that NAC (NAM‐ATAF1/2‐CUC2) transcription factor <jats:italic>SiNAC015</jats:italic> was a target of miR164b. The <jats:italic>sinac015</jats:italic> mutants showed attenuated drought tolerance, whereas overexpressing <jats:italic>mSiNAC015</jats:italic> (miR164b‐resistant version) improved drought tolerance in foxtail millet. Genetic evidence indicated that SiNAC015 could function in the same pathway as miR164b to mediate drought response by directly repressing the expression levels of <jats:italic>SitPRX</jats:italic> genes, which encoded peroxidase (POD) involved in reactive oxygen species (ROS) scavenging. Additionally, the superior <jats:italic>SiNAC015</jats:italic> <jats:sup> <jats:italic>Hap1</jats:italic> </jats:sup> possessing higher <jats:italic>SiNAC015</jats:italic> expression was found to be associated with enhanced drought tolerance in foxtail millet. Collectively, our study reveals that the miR164b‐ <jats:italic>SiNAC015</jats:italic> module mediates drought stress response and provides a valuable genetic resource for drought‐resistant breeding in foxtail millet.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"163 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145462003","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}
Yu Le, Meilin Chen, De Zhu, Zhiyong Xu, Chao Fu, Xinhui Xiong, Yuanxue Li, Ningyu Yang, Liuyang Hui, Xianlong Zhang, Zhongxu Lin
The allocation of carbon sources between cotton ovule and fibre significantly influences the yield and quality of seed and fibre. SWEET15 (Sugars Will Eventually be Exported Transporter 15) plays a key role in sucrose transport; however, the transcriptional regulation of SWEET15 in cotton remains unclear. Here, a SWEET15_A01, predominantly expressed during late ovule development (25–35 DPA), functions as a sucrose transporter between the ovule and fibre in cotton. Promoter variations between sea-island and upland cottons correlate with differential expression and cottonseed oil content. Overexpressing SWEET15_A01 in upland cotton reduces fibre length, seed size and oil content but increases lint percentage. Downregulation of SWEET15_A01 in upland cotton ovules upregulates other sugar transporters and cellulose synthase genes (CesAs) in fibres, indicating compensatory mechanisms. The R2R3-MYB transcription factor MYB44 directly binds to the SWEET15_A01 promoter, suppressing its expression, while bHLH3 interacts with MYB44 to weaken this repression. Overexpressing MYB44 increases fibre length but reduces seed size and oil content. This study reveals genetic variations for cottonseed oil improvement and elucidates how the MYB44/bHLH3-SWEET15_A01 module coordinates sugar allocation to balance seed and fibre development, offering strategies for enhancing cottonseed oil content.
{"title":"Transcriptional Regulation of SWEET15_A01 by MYB44/bHLH3 Modulates Carbon Allocation in Cotton Ovule and Fibre to Affect Seed and Fibre Traits","authors":"Yu Le, Meilin Chen, De Zhu, Zhiyong Xu, Chao Fu, Xinhui Xiong, Yuanxue Li, Ningyu Yang, Liuyang Hui, Xianlong Zhang, Zhongxu Lin","doi":"10.1111/pbi.70449","DOIUrl":"https://doi.org/10.1111/pbi.70449","url":null,"abstract":"The allocation of carbon sources between cotton ovule and fibre significantly influences the yield and quality of seed and fibre. SWEET15 (Sugars Will Eventually be Exported Transporter 15) plays a key role in sucrose transport; however, the transcriptional regulation of <i>SWEET15</i> in cotton remains unclear. Here, a <i>SWEET15_A01</i>, predominantly expressed during late ovule development (25–35 DPA), functions as a sucrose transporter between the ovule and fibre in cotton. Promoter variations between sea-island and upland cottons correlate with differential expression and cottonseed oil content. Overexpressing <i>SWEET15_A01</i> in upland cotton reduces fibre length, seed size and oil content but increases lint percentage. Downregulation of <i>SWEET15_A01</i> in upland cotton ovules upregulates other sugar transporters and cellulose synthase genes (<i>CesA</i>s) in fibres, indicating compensatory mechanisms. The R2R3-MYB transcription factor MYB44 directly binds to the <i>SWEET15_A01</i> promoter, suppressing its expression, while bHLH3 interacts with MYB44 to weaken this repression. Overexpressing MYB44 increases fibre length but reduces seed size and oil content. This study reveals genetic variations for cottonseed oil improvement and elucidates how the MYB44/bHLH3-SWEET15_A01 module coordinates sugar allocation to balance seed and fibre development, offering strategies for enhancing cottonseed oil content.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"77 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145454725","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}
Shoujian Zang, Dongjiao Wang, Liqian Qin, Shijiang Cui, Guran Wu, Kaisheng Liu, Qiugang Ding, Qianlong Hui, Tingting Sun, Yachun Su, Yingfang Zhu, Qibin Wu, Youxiong Que
<p>Sugarcane (<i>Saccharum</i> spp.) is essential for global sugar and bioenergy production, but its yield and quality are severely threatened by fungal diseases (Ling et al. <span>2025</span>). Plant defence against pathogens is primarily regulated by transcription factors (TFs) (Buscaill and Rivas <span>2014</span>), among which WRKYs can act as positive or negative immune regulators (Huang et al. <span>2022</span>). We previously reported that ScWRKY4 interacts with ScJAZ13 to suppress JA signalling and immune gene expression, increasing susceptibility to pathogens (Wang et al. <span>2024</span>). More recently, we found that ScWRKY2 reduces resistance to smut disease by interacting with the chloroplast protein ScPsbP and inducing ROS scavenging genes (Wang et al. <span>2025</span>). These findings indicate that WRKY TFs play diverse roles in sugarcane immunity. However, their contribution to immune homeostasis during fungal infection remains unclear.</p> <p>Here, we identified the sugarcane ScWRKY6, a class II-d WRKY TF, which contains two nuclear localization signals (NLSs), two nuclear export signals (NESs), a conserved zinc finger motif, and a WRKY domain (Figure 1A,B; Figure S1A–C; Table S1). Its expression is markedly induced by smut, pokkah boeng, and brown stripe diseases, suggesting its potential role in the broad-spectrum antifungal response of sugarcane (Figure 1C). Notably, ScWRKY6 was revealed as a nuclear protein that promotes intracellular ROS accumulation, as indicated by elevated ROS-scavenging gene expression and stronger H<sub>2</sub>DCF-DA (2′, 7′-dichlorodihydrofluorescein diacetate) fluorescence (Figure 1D,E; Figure S1D). These results suggest that ScWRKY6 may function as a regulator in sugarcane response to fungal pathogens.</p> <figure><picture> <source media="(min-width: 1650px)" srcset="/cms/asset/64187c1e-d3a9-46a5-bb54-307519710bab/pbi70444-fig-0001-m.jpg"/><img alt="Details are in the caption following the image" data-lg-src="/cms/asset/64187c1e-d3a9-46a5-bb54-307519710bab/pbi70444-fig-0001-m.jpg" loading="lazy" src="/cms/asset/6bfb6b74-8a84-4fc8-aba0-ee2e0c1a524b/pbi70444-fig-0001-m.png" title="Details are in the caption following the image"/></picture><figcaption> <div><strong>FIGURE 1<span style="font-weight:normal"></span></strong><div>Open in figure viewer<i aria-hidden="true"></i><span>PowerPoint</span></div> </div> <div>ScWRKY6-ScSAG39 module regulates <i>ScPR1</i> expression and immune homeostasis. (A) Conserved domains of the ScWRKY6 protein and its predicted NLS and NES. (B) 3D structural diagram of the ScWRKY6 protein. (C) Expression patterns of the <i>ScWRKY6</i> gene in response to smut, pokkah boeng, and brown stripe diseases. (D) Subcellular localization of the ScWRKY6-GFP fusion protein in sugarcane protoplasts. (E) The expression levels of genes involved in ROS scavenging after protoplast transfection (Student's <i>t</i>-test, <i>p</i> < 0.01). (F) Determination of the rice blast resistance of t
甘蔗（Saccharum spp.）对全球糖和生物能源生产至关重要，但其产量和质量受到真菌病害的严重威胁（Ling et al. 2025）。植物对病原体的防御主要由转录因子（transcription factors, TFs）调控（Buscaill and Rivas 2014），其中WRKYs可作为阳性或阴性免疫调节剂（Huang et al. 2022）。我们之前报道过ScWRKY4与ScJAZ13相互作用抑制JA信号和免疫基因表达，增加对病原体的易感性（Wang et al. 2024）。最近，我们发现ScWRKY2通过与叶绿体蛋白ScPsbP相互作用并诱导ROS清除基因来降低对黑穗病的抗性（Wang et al. 2025）。这些发现表明WRKY TFs在甘蔗免疫中发挥着不同的作用。然而，它们在真菌感染期间对免疫稳态的贡献尚不清楚。在这里，我们鉴定了甘蔗ScWRKY6，一个II-d类WRKY TF，它包含两个核定位信号（NLSs），两个核输出信号（NESs），一个保守的锌指基序和一个WRKY结构域（图1A，B；图S1A-C；表S1）。它的表达受黑穗病、白斑病和褐条病的显著诱导，这表明它在甘蔗的广谱抗真菌反应中具有潜在作用（图1C）。值得注意的是，ScWRKY6是一种促进细胞内ROS积累的核蛋白，ROS清除基因表达升高，H2DCF-DA（2 ', 7 ' -二氯二氢荧光素双乙酸酯）荧光增强（图1D，E；图S1D）。这些结果表明ScWRKY6可能在甘蔗对真菌病原体的反应中起调节作用。powerpointscwrky6 - scsag39模块调节ScPR1表达和免疫稳态。(A) ScWRKY6蛋白的保守结构域及其预测的NLS和NES。(B) ScWRKY6蛋白三维结构图。(C) ScWRKY6基因在黑穗病、白烟病和褐条病中的表达模式。(D) ScWRKY6-GFP融合蛋白在甘蔗原生质体中的亚细胞定位。(E)转染原生质体后参与活性氧清除的基因表达水平（学生t检验，p < 0.01）。(F)喷雾法测定scwrky6过表达系的稻瘟病抗性。(G) WT和scwrky6过表达植株的疾病反应和H2O2积累。(H) WT和scwrky6过表达植株病斑传播长度的统计分析（学生t检验，p < 0.05, n = 3）。(1) M. oryzae感染前后WT和scwrky6过表达植株中OsPR1的表达水平。不同字母表示差异显著（学生t检验，p < 0.05, n = 3）。(J)在EMSA分析中，ScWRKY6的WRKY结构域直接与W-box元件的探针1结合，而不与探针2结合。(K)在酵母单杂交实验中，ScWRKY6与ScPR1启动子P1片段直接相关。(L)双荧光素酶表达分析显示ScWRKY6转录激活了ScPR1启动子活性。(M)不同浓度ScSAG39-myc对ScWRKY6-GFP亚细胞定位的影响。(N)不同浓度ScSAG39-myc表达scwrky6 - gfp细胞的核数定量（学生t检验，****p < 0.0001）。(O)不同浓度ScWRKY6-GFP共浸润ScSAG39-myc后蛋白水平的测定。(P) EMSA实验中ScSAG39蛋白对ScWRKY6与ScPR1启动子结合能力的影响。(Q) ScSAG39抑制scwrky6介导的ScPR1转录激活。(R)该模型说明了ScWRKY6与ScSAG39相互作用调节免疫应答的机制。随后，我们建立了水稻(Oryza sativa) -Magnaporthe oryzae系统，利用遗传和生化方法研究ScWRKY6在真菌抗病中的调节作用（图S1E-G）。有趣的是，接种M. oryzae菌株Guy11后，ScWRKY6-OE转基因植株比野生型（WT）植株的病变更少，H2O2积累显著增加（图1F，G）。在穿孔接种试验中，ScWRKY6-OE植株的损伤长度明显短于WT植株（图1G，H）。此外，接种后48 h，防御相关基因OsPR1在ScWRKY6- oe植株中的表达水平显著升高（图1I），表明ScWRKY6可以正向调节水稻对M. oryzae的抗性。为了阐明ScWRKY6在水稻抗稻瘟病中的作用，我们对接种(T)或未接种M. oryzae （CK）的WT和ScWRKY6- oe1植株的12个cDNA文库进行了rna测序（图S2A；表S2）。在WT-CK_vs_WT-T和ScWRKY6-CK_vs_ScWRKY6-T组中分别鉴定出4947和1838个deg（图S2B-E）。 其中，WT-CK_vs_WT-T上调基因主要与基础代谢和生物合成途径相关。相比之下，ScWRKY6-CK_vs_ScWRKY6-T中上调的基因主要富集于与植物免疫应答相关的通路中（图S2F、G；表S3）。有趣的是，ScWRKY6过表达激活了苯丙素生物合成途径，大多数相关的deg上调，包括PAL、C4H和4CL等关键酶（图S3）。令人惊讶但合理的是，许多TFs和抗性(R)基因，如NLR-、RLK-和wak型基因，在ScWRKY6- oe系中特异性上调（图S4；表S4和表S5），表明ScWRKY6通过促进次生代谢和R基因表达来增强水稻对M. oryzae的防御。此外，我们发现ScWRKY6可以上调OsPR1的表达，介导植物的免疫应答（图1I）。为了确定其是否直接调控PR1，我们从甘蔗中克隆了具有转录活性并可被甘蔗孢菌感染诱导的同源基因ScPR1的启动子（图S5A-C）。该启动子包含两个W-box基序，其中一个（probe1）被ScWRKY6特异性结合（图1J，K；图S5D）。双荧光素酶和ChIP-qPCR进一步证实ScWRKY6可以激活ScPR1的转录（图1L；图S6）。这些结果表明，ScWRKY6直接与ScPR1启动子结合，很可能参与了病原体诱导的ScPR1表达。为了鉴定参与疾病应答的潜在scwrky6相互作用蛋白，使用来自黑穗病感染芽的甘蔗cDNA文库进行酵母双杂交（Y2H）筛选（图S7A）。它们主要参与阿拉伯糖代谢、蛋白质降解和半胱氨酸蛋白酶活性（图S7B）。其中，我们选择ScSAG39进行进一步的研究，ScSAG39是一种半胱氨酸蛋白酶基因，该基因在S. scitamineum感染后下调（图S8A-C）。ScSAG39在烟叶中的过表达加重了疾病症状，减少了H2O2的积累，暗示在植物防御中起负作用。ROS-和hr相关基因的表达改变进一步支持了这一点（图S8D-F）。我们还发现ScSAG39是一种内质网相关蛋白（图S8G）。Y2H、BiFC和Co-IP检测证实了其与ScWRKY6的相互作用（图S9）。值得注意的是，共表达实验显示ScSAG39丰度的变化改变了ScWRKY6的核定位（图1M-O）。ScSAG39限制了ScWRKY6的核输入，降低ScSAG39水平允许更多的ScWRKY6在细胞核中积累（图1N；图S9A）。此外，ScSAG39还通过竞争性阻断ScWRKY6与W-box元件的结合来抑制ScPR1的激活（图1P），其与ScWRKY6的共表达导致ScPR1启动子驱动的荧光素酶活性显著降低（图1Q；图S10）。此外，ScSAG39和W-
{"title":"ScWRKY6 Interacts With ScSAG39 to Regulate Immune Homeostasis by Transcriptional Control of ScPR1","authors":"Shoujian Zang, Dongjiao Wang, Liqian Qin, Shijiang Cui, Guran Wu, Kaisheng Liu, Qiugang Ding, Qianlong Hui, Tingting Sun, Yachun Su, Yingfang Zhu, Qibin Wu, Youxiong Que","doi":"10.1111/pbi.70444","DOIUrl":"https://doi.org/10.1111/pbi.70444","url":null,"abstract":"<p>Sugarcane (<i>Saccharum</i> spp.) is essential for global sugar and bioenergy production, but its yield and quality are severely threatened by fungal diseases (Ling et al. <span>2025</span>). Plant defence against pathogens is primarily regulated by transcription factors (TFs) (Buscaill and Rivas <span>2014</span>), among which WRKYs can act as positive or negative immune regulators (Huang et al. <span>2022</span>). We previously reported that ScWRKY4 interacts with ScJAZ13 to suppress JA signalling and immune gene expression, increasing susceptibility to pathogens (Wang et al. <span>2024</span>). More recently, we found that ScWRKY2 reduces resistance to smut disease by interacting with the chloroplast protein ScPsbP and inducing ROS scavenging genes (Wang et al. <span>2025</span>). These findings indicate that WRKY TFs play diverse roles in sugarcane immunity. However, their contribution to immune homeostasis during fungal infection remains unclear.</p>\u0000<p>Here, we identified the sugarcane ScWRKY6, a class II-d WRKY TF, which contains two nuclear localization signals (NLSs), two nuclear export signals (NESs), a conserved zinc finger motif, and a WRKY domain (Figure 1A,B; Figure S1A–C; Table S1). Its expression is markedly induced by smut, pokkah boeng, and brown stripe diseases, suggesting its potential role in the broad-spectrum antifungal response of sugarcane (Figure 1C). Notably, ScWRKY6 was revealed as a nuclear protein that promotes intracellular ROS accumulation, as indicated by elevated ROS-scavenging gene expression and stronger H<sub>2</sub>DCF-DA (2′, 7′-dichlorodihydrofluorescein diacetate) fluorescence (Figure 1D,E; Figure S1D). These results suggest that ScWRKY6 may function as a regulator in sugarcane response to fungal pathogens.</p>\u0000<figure><picture>\u0000<source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/64187c1e-d3a9-46a5-bb54-307519710bab/pbi70444-fig-0001-m.jpg\"/><img alt=\"Details are in the caption following the image\" data-lg-src=\"/cms/asset/64187c1e-d3a9-46a5-bb54-307519710bab/pbi70444-fig-0001-m.jpg\" loading=\"lazy\" src=\"/cms/asset/6bfb6b74-8a84-4fc8-aba0-ee2e0c1a524b/pbi70444-fig-0001-m.png\" title=\"Details are in the caption following the image\"/></picture><figcaption>\u0000<div><strong>FIGURE 1<span style=\"font-weight:normal\"></span></strong><div>Open in figure viewer<i aria-hidden=\"true\"></i><span>PowerPoint</span></div>\u0000</div>\u0000<div>ScWRKY6-ScSAG39 module regulates <i>ScPR1</i> expression and immune homeostasis. (A) Conserved domains of the ScWRKY6 protein and its predicted NLS and NES. (B) 3D structural diagram of the ScWRKY6 protein. (C) Expression patterns of the <i>ScWRKY6</i> gene in response to smut, pokkah boeng, and brown stripe diseases. (D) Subcellular localization of the ScWRKY6-GFP fusion protein in sugarcane protoplasts. (E) The expression levels of genes involved in ROS scavenging after protoplast transfection (Student's <i>t</i>-test, <i>p</i> < 0.01). (F) Determination of the rice blast resistance of t","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"109 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145454724","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}
In bread wheat (Triticum aestivum L.), cysteine (Cys) residues within the N- and C-termini of high-molecular-weight glutenin subunits (HMW-GSs) critically influence dough quality. However, the functional significance of Cys residue in their central repeat domain (CRD) remains unclear. Using site-directed mutagenesis (SDM), we introduced Cys residues near the N-terminus (m1) and/or C-terminus (m2) of 1Dx2 CRD, generating variants 1Dx2m1, 1Dx2m2 and 1Dx2m1/2. Transgenic lines expressing the variants exhibited superior dough properties, increased loaf volume, and elevated glutenin macropolymers (GMPs) content, attributable to enhanced disulfide bond formation and upregulation of associated genes. Notably, the two Cys residues introduced variant 1Dx2m1/2 demonstrated additive improvements, indicating synergistical effects of Cys residues at both positions. Field trials confirmed these modifications did not compromise key agronomic traits. Our study provides the experimental evidence for the role of CRD-located Cys residues in HMW-GSs on dough quality and offers valuable genetic resources for improving end-use quality without yield penalties in wheat breeding.
{"title":"Improving End-Use Quality Through Introducing Cysteines Into the Central Repeat Domain of High-Molecular-Weight Glutenin Subunit 1Dx2 in Bread Wheat","authors":"Bo Wei, Lujun Zhang, Renchun Fan, Qi Zhang, Jiazhu Sun, Changbin Yin, Dongcheng Liu, Xiukun Liu, Aifeng Liu, Yuling Jiao, Caixia Gao, Xu Jia, Xianping Wang, Aimin Zhang, Xinyou Cao, Daowen Wang, Xiangqi Zhang","doi":"10.1111/pbi.70451","DOIUrl":"https://doi.org/10.1111/pbi.70451","url":null,"abstract":"In bread wheat (<i>Triticum aestivum</i> L<i>.</i>), cysteine (Cys) residues within the N- and C-termini of high-molecular-weight glutenin subunits (HMW-GSs) critically influence dough quality. However, the functional significance of Cys residue in their central repeat domain (CRD) remains unclear. Using site-directed mutagenesis (SDM), we introduced Cys residues near the N-terminus (m1) and/or C-terminus (m2) of 1Dx2 CRD, generating variants <i>1Dx2m1</i>, <i>1Dx2m2</i> and <i>1Dx2m1/2</i>. Transgenic lines expressing the variants exhibited superior dough properties, increased loaf volume, and elevated glutenin macropolymers (GMPs) content, attributable to enhanced disulfide bond formation and upregulation of associated genes. Notably, the two Cys residues introduced variant <i>1Dx2m1/2</i> demonstrated additive improvements, indicating synergistical effects of Cys residues at both positions. Field trials confirmed these modifications did not compromise key agronomic traits. Our study provides the experimental evidence for the role of CRD-located Cys residues in HMW-GSs on dough quality and offers valuable genetic resources for improving end-use quality without yield penalties in wheat breeding.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145455193","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}
To develop salt‐tolerant transgenic soybeans, ScHAL1 from Saccharomyces cerevisiae (yeast), which increases salt tolerance by maintaining high intracellular K + concentrations and decreasing intracellular Na + during salt stress, was introduced into soybean ( Glycine max L. Merr.) through Agrobacterium tumefaciens ‐mediated transformation. Among the 373 transgenic plants generated, three lines (TL1, TL2 and TL3) exhibiting stable and enhanced salt tolerance via ScHAL1‐mediated ion homeostasis were selected. Molecular analyses of the transgenic lines by polymerase chain reaction (PCR), Southern blotting, reverse transcription‐PCR (RT‐PCR), quantitative reverse‐transcription polymerase chain reaction (qRT‐PCR), western blotting and enzyme‐linked immunosorbent assay (ELISA) showed that ScHAL1 and bar transgenes were stably inherited and expressed across multiple generations (T4, T5 and T6). Under 300 mM NaCl salt stress, the transgenic lines TL1, TL2 and TL3 exhibited enhanced salt tolerance, which was stably inherited by their progenies, as well as improved agronomic traits under salt stress, with an average yield reduction of only 8.61%, compared with 34.8% in non‐transgenic (NT) control plants. Under salt stress, physiological indices of the transgenic lines showed that ScHAL1 expressed in soybean synergizes with analogous ion transporters to stabilise cytoplasmic K + /Na + ratios by reducing Na + influx and promoting K + retention to limit cytosolic Na + toxicity. TL1, exhibiting stable and enhanced salt tolerance through ScHAL1 overexpression, has been approved for environmental release and is currently undergoing biosafety assessment in pre‐production field trials. The ScHAL1 ‐overexpressing lines show potential for incorporation into commercial soybean breeding programs aimed at improving salinity tolerance in elite cultivars.
为了培育耐盐转基因大豆，通过农杆菌介导的转化，将来自酿酒酵母（Saccharomyces cerevisiae，酵母）的ScHAL1基因导入大豆（Glycine max L. Merr.），该基因在盐胁迫下通过维持高细胞内K +浓度和降低细胞内Na +浓度来提高耐盐性。在373个转基因植株中，选择了3个通过ScHAL1介导的离子稳态表现出稳定和增强的耐盐性的株系（TL1、TL2和TL3）。通过聚合酶链反应（PCR）、Southern blotting、逆转录- PCR （RT - PCR）、定量逆转录-聚合酶链反应（qRT - PCR）、western blotting和酶联免疫吸附试验（ELISA）对转基因系进行分子分析，结果表明ScHAL1和bar基因在多代（T4、T5和T6）中稳定遗传和表达。在300 mM NaCl盐胁迫下，转基因株系TL1、TL2和TL3表现出较强的耐盐性，并稳定地遗传给后代，在盐胁迫下的农艺性状得到改善，平均产量仅下降8.61%，而非转基因对照株系（NT）平均产量下降34.8%。盐胁迫下，转基因大豆株系的生理指标表明，大豆中表达的ScHAL1与类似的离子转运蛋白协同作用，通过减少Na +内流和促进K +滞留来稳定胞质K + /Na +比率，从而限制胞质Na +毒性。TL1通过ScHAL1过表达表现出稳定和增强的耐盐性，已被批准环境释放，目前正在进行生产前田间试验的生物安全性评估。这些过表达ScHAL1‐的品系显示了将其纳入旨在提高优良品种耐盐性的商业大豆育种计划的潜力。
{"title":"ScHAL1 ‐Mediated Enhancement of Salt Tolerance in Soybean: From Stable Transgenic Inheritance to Field Trial Validation","authors":"Zhijing Yu, Jia Wei, Qinan Cai, Yuanyu Zhang, Qianqian Zhao, Lu Niu, Pengfei Ren, Lixia Shang, Xiangdong Yang, Yingshan Dong, Rui Ma","doi":"10.1111/pbi.70452","DOIUrl":"https://doi.org/10.1111/pbi.70452","url":null,"abstract":"To develop salt‐tolerant transgenic soybeans, <jats:italic>ScHAL1</jats:italic> from <jats:styled-content style=\"fixed-case\"> <jats:italic>Saccharomyces cerevisiae</jats:italic> </jats:styled-content> (yeast), which increases salt tolerance by maintaining high intracellular K <jats:sup>+</jats:sup> concentrations and decreasing intracellular Na <jats:sup>+</jats:sup> during salt stress, was introduced into soybean ( <jats:styled-content style=\"fixed-case\"> <jats:italic>Glycine max</jats:italic> </jats:styled-content> L. Merr.) through <jats:styled-content style=\"fixed-case\"> <jats:italic>Agrobacterium tumefaciens</jats:italic> </jats:styled-content> ‐mediated transformation. Among the 373 transgenic plants generated, three lines (TL1, TL2 and TL3) exhibiting stable and enhanced salt tolerance via ScHAL1‐mediated ion homeostasis were selected. Molecular analyses of the transgenic lines by polymerase chain reaction (PCR), Southern blotting, reverse transcription‐PCR (RT‐PCR), quantitative reverse‐transcription polymerase chain reaction (qRT‐PCR), western blotting and enzyme‐linked immunosorbent assay (ELISA) showed that <jats:italic>ScHAL1</jats:italic> and <jats:italic>bar</jats:italic> transgenes were stably inherited and expressed across multiple generations (T4, T5 and T6). Under 300 mM NaCl salt stress, the transgenic lines TL1, TL2 and TL3 exhibited enhanced salt tolerance, which was stably inherited by their progenies, as well as improved agronomic traits under salt stress, with an average yield reduction of only 8.61%, compared with 34.8% in non‐transgenic (NT) control plants. Under salt stress, physiological indices of the transgenic lines showed that ScHAL1 expressed in soybean synergizes with analogous ion transporters to stabilise cytoplasmic K <jats:sup>+</jats:sup> /Na <jats:sup>+</jats:sup> ratios by reducing Na <jats:sup>+</jats:sup> influx and promoting K <jats:sup>+</jats:sup> retention to limit cytosolic Na <jats:sup>+</jats:sup> toxicity. TL1, exhibiting stable and enhanced salt tolerance through <jats:italic>ScHAL1</jats:italic> overexpression, has been approved for environmental release and is currently undergoing biosafety assessment in pre‐production field trials. The <jats:italic>ScHAL1</jats:italic> ‐overexpressing lines show potential for incorporation into commercial soybean breeding programs aimed at improving salinity tolerance in elite cultivars.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"2 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145447186","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}
Su‐Kyoung Lee, Woo‐Jong Hong, Eui‐Jung Kim, Choonseok Lee, Sunok Moon, Sun‐Hwa Ha, Ki‐Hong Jung
Efficient production of hybrid rice seeds requires male sterility systems to overcome the challenges of self‐pollination. In this study, we identified male gametic transfer defect ( GTrD ) 5 and GTrD9 as essential genes for the process of male gamete transmission in rice. Mutations in these genes cause partial male sterility by impairing pollen tube elongation, thereby reducing the fertilisation rate. Using CRISPR/Cas9 technology, we developed gtrd5flo5 and gtrd9flo5 double mutants that combined the gtrd5 and gtrd9 genes responsible for partial male sterility with the floury endosperm (FLO)5 gene, whose defects result in seeds with an opaque endosperm, allowing us to easily differentiate between hybrid and self‐pollinated seeds. This two‐line hybrid system demonstrated a high rate of hybrid seed production with significantly reduced self‐pollination ratios. The hybrid seeds resulted in plants with increased height, panicle size and grain yield compared with those obtained from the parental lines and also displayed heterosis. Unlike the current two‐line hybrid systems based on photoperiod‐ and thermosensitive genic male‐sterile lines, our approach is independent of environmental factors, ensuring a stable and reliable hybrid seed production. This novel method simplifies seed production, enhances efficiency and offers a cost‐effective and environmentally sustainable solution for hybrid rice breeding.
{"title":"Developing an Efficient System for Hybrid Rice Seed Production Using Partial Male Sterility","authors":"Su‐Kyoung Lee, Woo‐Jong Hong, Eui‐Jung Kim, Choonseok Lee, Sunok Moon, Sun‐Hwa Ha, Ki‐Hong Jung","doi":"10.1111/pbi.70448","DOIUrl":"https://doi.org/10.1111/pbi.70448","url":null,"abstract":"Efficient production of hybrid rice seeds requires male sterility systems to overcome the challenges of self‐pollination. In this study, we identified male <jats:italic>gametic transfer defect</jats:italic> ( <jats:italic>GTrD</jats:italic> ) <jats:italic>5</jats:italic> and <jats:italic>GTrD9</jats:italic> as essential genes for the process of male gamete transmission in rice. Mutations in these genes cause partial male sterility by impairing pollen tube elongation, thereby reducing the fertilisation rate. Using CRISPR/Cas9 technology, we developed <jats:italic>gtrd5flo5</jats:italic> and <jats:italic>gtrd9flo5</jats:italic> double mutants that combined the <jats:italic>gtrd5</jats:italic> and <jats:italic>gtrd9</jats:italic> genes responsible for partial male sterility with the <jats:italic>floury endosperm (FLO)5</jats:italic> gene, whose defects result in seeds with an opaque endosperm, allowing us to easily differentiate between hybrid and self‐pollinated seeds. This two‐line hybrid system demonstrated a high rate of hybrid seed production with significantly reduced self‐pollination ratios. The hybrid seeds resulted in plants with increased height, panicle size and grain yield compared with those obtained from the parental lines and also displayed heterosis. Unlike the current two‐line hybrid systems based on photoperiod‐ and thermosensitive genic male‐sterile lines, our approach is independent of environmental factors, ensuring a stable and reliable hybrid seed production. This novel method simplifies seed production, enhances efficiency and offers a cost‐effective and environmentally sustainable solution for hybrid rice breeding.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"39 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145447187","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}
Menglong Fan, Hong Jiang, Yuxiao Qu, Ying Zhang, Xinlei Li, Yan Wang
The role of transposable elements (TEs) in genome evolution and phenotypic diversification in Camellia remains poorly understood. Here, we present an integrated analysis of genome resequencing data from 237 Camellia accessions and 11 de novo genome assemblies representing all major floral colour types. We constructed a comprehensive phylogenetic framework for the genus and suggest that the most recent common ancestor likely had white flowers. Comparative genomic analyses reveal structural variants across species that overlap with numerous transposable elements and contribute to genome content divergence. Using a graph‐based genome to characterise these structural variants, we find that lineage‐specific TE amplifications drive the regulatory network rewiring, which modulates homoeologous gene expression, influencing flower colour diversification. Further experimental validation identifies a lineage‐specific, high‐frequency presence variation mediated by a TIR transposon that regulates MYB60 expression, suppressing anthocyanin biosynthesis and leading to large‐scale floral colour divergence. Therefore, these findings highlight the central role of TE‐mediated regulatory innovation in the evolution of flower colour in Camellia and offer broader insights into the molecular mechanisms driving phenotypic diversification in plants.
{"title":"Transposable Element‐Mediated Structural Variation Drives Flower Colour Diversification in Camellia","authors":"Menglong Fan, Hong Jiang, Yuxiao Qu, Ying Zhang, Xinlei Li, Yan Wang","doi":"10.1111/pbi.70442","DOIUrl":"https://doi.org/10.1111/pbi.70442","url":null,"abstract":"The role of transposable elements (TEs) in genome evolution and phenotypic diversification in <jats:italic>Camellia</jats:italic> remains poorly understood. Here, we present an integrated analysis of genome resequencing data from 237 <jats:italic>Camellia</jats:italic> accessions and 11 de novo genome assemblies representing all major floral colour types. We constructed a comprehensive phylogenetic framework for the genus and suggest that the most recent common ancestor likely had white flowers. Comparative genomic analyses reveal structural variants across species that overlap with numerous transposable elements and contribute to genome content divergence. Using a graph‐based genome to characterise these structural variants, we find that lineage‐specific TE amplifications drive the regulatory network rewiring, which modulates homoeologous gene expression, influencing flower colour diversification. Further experimental validation identifies a lineage‐specific, high‐frequency presence variation mediated by a TIR transposon that regulates <jats:italic>MYB60</jats:italic> expression, suppressing anthocyanin biosynthesis and leading to large‐scale floral colour divergence. Therefore, these findings highlight the central role of TE‐mediated regulatory innovation in the evolution of flower colour in <jats:italic>Camellia</jats:italic> and offer broader insights into the molecular mechanisms driving phenotypic diversification in plants.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"38 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145447185","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}
Marissa K. Simon, Li Yuan, Ping Che, Kevin Day, Todd Jones, Ian D. Godwin, Anna M. G. Koltunow, Marc C. Albertsen
Induction of apomixis, or clonal reproduction through seed, could economise commercial hybrid seed production and enable smallholder farmers to save and sow hybrid seed. Here, we demonstrate the synthetic induction of apomixis in two sorghum hybrids and show that the clonal hybrid seed can be maintained across multiple seed generations. This was achieved through the combination of avoidance of meiosis and induced parthenogenesis. Meiotic avoidance was generated by CRISPR/Cas9 knockout of the sorghum meiosis genes Spo11, Rec8, OsdL1 and OsdL3. Parthenogenesis was induced in the resultant diploid egg cell by the expression of the Cenchrus ASGR-BBML2 gene coding sequence. Two different strategies were used to combine these components to induce synthetic apomixis in two sorghum hybrids. Each hybrid used Tx623 as a female parent and either Tx430 or the African landrace Macia as a male parent. Seed yields in both apomictic hybrids were consistent and stable for multiple generations following self-pollination but reduced relative to the sexual hybrids. Sorghum contains two copies of the Osd1 gene that function in meiotic non-reduction. CRISPR/Cas9 knockout of both OsdL1 and OsdL3 loci was sufficient to produce clonal hybrid progeny in conjunction with the other components, but this led to a reduction in seed set. By contrast, a single in-frame edited allele of either OsdL1 or OsdL3 significantly improved seed set of clonal hybrid progeny. Fine-tuning OsdL activity appears to be essential to optimising fertility; however, additional improvements are required to unlock the agronomic potential of synthetically induced apomictic sorghum in the field.
{"title":"Induction of Synthetic Apomixis in Two Sorghum Hybrids Enables Seed Yield and Genotype Preservation Over Multiple Generations","authors":"Marissa K. Simon, Li Yuan, Ping Che, Kevin Day, Todd Jones, Ian D. Godwin, Anna M. G. Koltunow, Marc C. Albertsen","doi":"10.1111/pbi.70441","DOIUrl":"https://doi.org/10.1111/pbi.70441","url":null,"abstract":"Induction of apomixis, or clonal reproduction through seed, could economise commercial hybrid seed production and enable smallholder farmers to save and sow hybrid seed. Here, we demonstrate the synthetic induction of apomixis in two sorghum hybrids and show that the clonal hybrid seed can be maintained across multiple seed generations. This was achieved through the combination of avoidance of meiosis and induced parthenogenesis. Meiotic avoidance was generated by CRISPR/Cas9 knockout of the sorghum meiosis genes <i>Spo11</i>, <i>Rec8</i>, <i>OsdL1</i> and <i>OsdL3</i>. Parthenogenesis was induced in the resultant diploid egg cell by the expression of the <i>Cenchrus ASGR-BBML2</i> gene coding sequence. Two different strategies were used to combine these components to induce synthetic apomixis in two sorghum hybrids. Each hybrid used Tx623 as a female parent and either Tx430 or the African landrace Macia as a male parent. Seed yields in both apomictic hybrids were consistent and stable for multiple generations following self-pollination but reduced relative to the sexual hybrids. Sorghum contains two copies of the <i>Osd1</i> gene that function in meiotic non-reduction. CRISPR/Cas9 knockout of both <i>OsdL1</i> and <i>OsdL3</i> loci was sufficient to produce clonal hybrid progeny in conjunction with the other components, but this led to a reduction in seed set. By contrast, a single in-frame edited allele of either <i>OsdL1</i> or <i>OsdL3</i> significantly improved seed set of clonal hybrid progeny. Fine-tuning <i>OsdL</i> activity appears to be essential to optimising fertility; however, additional improvements are required to unlock the agronomic potential of synthetically induced apomictic sorghum in the field.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"90 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145440904","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}
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