<p>Cabbage (<i>Brassica oleracea</i> L. var. <i>capitata</i>) is one of the most important vegetables grown globally (Li et al. <span>2024</span>). Currently, more than 90% of cabbage cultivars are male sterile. Male sterility-based hybridisation facilitates heterosis utilisation and yield enhancement (Han et al. <span>2023</span>). Ogura cytoplasmic male sterility (CMS) is more commonly used due to its maternal inheritance and easy transferability (Ren et al. <span>2020</span>, <span>2022</span>). However, CMS varieties cannot be self-pollinated, thereby limiting germplasm innovation and restricting genetic diversity, which can be solved by fertility restorer lines (Li et al. <span>2021</span>; Ren et al. <span>2022</span>). The <i>Rfo</i> gene exists only in radish but not in any <i>B. oleracea</i> crops. This gene has been transferred from radish into rapeseed, a related species of <i>B. oleracea</i>, providing a promising resource for creating cabbage Ogura CMS fertility restorer lines (FRLs) (Yu et al. <span>2020</span>). In our previous studies, <i>Rfo</i> was successfully transferred from rapeseed into Chinese kale and cabbage through distant hybridisation (Yu et al. <span>2020</span>; Ren et al. <span>2020</span>). However, critical knowledge gaps remain regarding the genome composition and genetic effects of the cabbage Ogura CMS restorer lines.</p> <p>Initially, the absence of <i>Rfo</i>-homozygous lines and high heterozygosity hindered <i>de novo</i> genome assembly of Ogura CMS FRLs (Table S1). In this study, BC<sub>7</sub> high-generation Ogura CMS FRLs were developed via successive backcrossing (Figure 1A). To overcome this challenge, we generated a chromosome-level genome assembly for the BC<sub>7</sub> FRL individual RFO7GH by combining Oxford Nanopore ultra-long reads, Illumina short reads for error correction and Hi-C scaffolding for chromosomal phasing (Figure S1A; Figure S2; Table S2). The <i>de novo</i> assembly yielded 40 contigs (N50 = 32.3 Mb) spanning 601.8 Mb from 59 Gb raw data. The final assembly contains more than 98.3% of the genome sequences (591.8 Mb) anchored to nine pseudochromosomes. Crucially, a 15.8-Mb alien radish genomic fragment (R09: 0–15 789 821 bp) containing the <i>Rfo</i> gene was inverted and integrated into the long-arm terminal region of C09 in RFO7GH, replacing the homologous 28.2-Mb cabbage genomic fragment (C09: 0–28 239 208 bp) (Figure 1B). Genome analysis revealed the loss of the telomere on the C09 long arm (Figure 1C). To validate this structural rearrangement, a specific primer set (GR-4F/4R) targeting the recombination breakpoint was designed, which successfully amplified a 4.6-kb junction sequence (Table S3). Sanger sequencing of the amplified products showed 100% concordance with Nanopore data, confirming the assembly accuracy.</p> <figure><picture> <source media="(min-width: 1650px)" srcset="/cms/asset/acaa9a00-9101-4b30-9f96-e267159a9b61/pbi70396-fig-0001-m.jpg"/><img alt="Details are i
白菜（芸苔甘蓝L. var. capitata）是全球种植的最重要的蔬菜之一（Li et al. 2024）。目前，超过90%的卷心菜品种是雄性不育的。基于雄性不育的杂交有利于杂种优势的利用和产量的提高（Han et al. 2023）。小谷细胞质雄性不育（Ogura cytoplasmic male infertility， CMS）因其母系遗传和易于转移而更常用（Ren et al. 2020, 2022）。但是，CMS品种不能自花授粉，从而限制了种质创新，限制了遗传多样性，这可以通过育性恢复系来解决（Li et al. 2021; Ren et al. 2022）。Rfo基因仅存在于萝卜中，不存在于任何甘蓝作物中。该基因已从萝卜转移到甘蓝的亲缘种油菜籽中，为创建白菜小白菜CMS育性恢复系（frl）提供了有希望的资源（Yu et al. 2020）。在我们之前的研究中，Rfo通过远缘杂交成功地从油菜籽转移到芥蓝和白菜中（Yu et al. 2020; Ren et al. 2020）。然而，关于小白菜CMS恢复系的基因组组成和遗传效应的关键知识差距仍然存在。最初，缺乏rfo纯合子系和高杂合性阻碍了Ogura CMS frl的从头基因组组装（表S1）。本研究通过连续回交培育BC7高代小仓CMS frl（图1A）。为了克服这一挑战，我们结合Oxford Nanopore超长reads、Illumina短reads进行错误校正和Hi-C scaffolding进行染色体分相，为BC7 FRL个体RFO7GH构建了染色体水平的基因组组装（图S1A；图S2；表S2）。重新组装产生40个contigs (N50 = 32.3 Mb)，从59 Gb原始数据中跨越601.8 Mb。最终组装包含超过98.3%的基因组序列（591.8 Mb），锚定在9条假染色体上。重要的是，含有Rfo基因的15.8 mb外源萝卜基因组片段（R09: 0-15 789 821 bp）被倒置并整合到RFO7GH中C09的长臂末端区域，取代了同源的28.2 mb白菜基因组片段（C09: 0-28 239 208 bp）（图1B）。基因组分析显示C09长臂上的端粒缺失（图1C）。为了验证这种结构重排，设计了针对重组断点的特定引物集（GR-4F/4R），成功扩增了4.6 kb的连接序列（表S3）。扩增产物的Sanger测序结果与Nanopore数据100%一致，证实了组装的准确性。图1基于高代小仓CMS frl的无外来遗传背景干扰的小仓CMS育性恢复系统。(A)建立生育恢复系统和开发携带CRa基因的抗大白菜的路线图。(B) RFO7GH rfo -负单倍型（Bol_1）和rfo -正单倍型（Bol_2）与萝卜Rapsa_Xiang_V1.0参考基因组比对（http://brassicadb.org）。红框表示15.8 mb外来萝卜基因组片段比对结果。(C)正常C09（上）和rfo阳性C09（下）示意图。红框表示15.8 mb的外来萝卜基因组片段。灰色框代表卷心菜端粒，黄色框代表萝卜端粒。我们的研究不仅恢复了Ogura CMS品种的育性，而且还显示了端粒功能障碍和染色体倒转带来的三个新好处，即在细胞质替换后，无需额外的标记辅助选择，就可以快速丢失外源基因组片段：(i)传播屏障确保了外源片段的快速丢失：对285个群体（n = 51 787）的多代分析显示，Rfo基因存在严重的传播障碍。自花授粉群体的平均传粉率为8.05%，回交群体的平均传粉率为1.18%（表S1）。使用外源萝卜片段特异性的9 kb探针对减数分裂前期I细胞进行荧光原位杂交（FISH），结果显示，只有非常小比例的pmc（29/ 412,7%）携带外源基因组片段（图S3；表S3）。大的染色体倒位和至关重要的端粒丢失似乎严重破坏了基因组的稳定性，使携带15.8 mb外源片段的细胞难以进行减数分裂并产生配子，从而导致快速丢失。（ii）完整的外源片段实现一步清除：C09杂合性从BC1（63%）逐渐下降到BC4 (38%)， BC4后稳定，完全抑制重组（图S1B）。设计了26对引物（BoRa1p-BoRa26p），目标序列均匀分布在15.8 mb的外来萝卜基因组片段中（表S3）。 这些引物与断点特异性引物GR-4F/4R一起用于对所有rfo阳性个体进行基因分型。基因分型证实，15.8 mb的外源萝卜片段是所有Ogura CMS frl中与育性恢复完全共分离的最短外源片段（图S4），并且在所有rfo阳性个体中作为一个完整的单元遗传。这种完整性与传输屏障（优势i）相结合，使整个外来碎片能够迅速丢失。对5000多名rfo阴性个体的筛选证实，没有人含有任何外源萝卜片段，进一步证明了外源萝卜片段的快速丢失（图S4）。（iii）生长惩罚提供了万无一失的消除外来碎片携带者：没有外来碎片的个体表现出更强的生长潜力。rfo阳性个体外叶长（LL）和经济产量（EY）显著降低，外叶宽（LW）显著降低（图S1C，D）。虽然外源片段遗传给后代的情况很少（~1%），但由于农艺表现不佳，携带外源片段的个体在选择育种中很容易被淘汰。通过在7个携带棍棒病（CRa）、黑腐病（Xcc3）和枯萎病（Foc1）抗性基因的遗传多样性品种中恢复育性，验证了该系统的有效性。以含cra种质为例，在BC1进行育性恢复和细胞质置换后，从11个组合中获得1044个cra阳性个体（n = 2880）（表S4；图S1E）。在这些个体中，有59个个体表现出显著的生物量减少（23.4%-38.1%,p < 0.001），在选择过程中被表型淘汰（表S4）。综合筛选显示，15.8 mb的片段仅保留28/1044；所有这28个个体都是先前仅基于表型筛选而被淘汰的59个个体的一部分。这证实了该系统仅通过表型筛选清除外来片段的效率。总之，本研究通过远缘杂交和连续回交获得了高代小稻CMS frl。重新组装了高质量的FRL系RFO7GH基因组。构建了一套无外源片段干扰的小白菜育性恢复系统。小仓CMS frl可使雄性不育抗病品种转化为雄性可育种质，在白菜育种中具有一定的应用价值。
{"title":"A 15.8-Mb Alien Radish Chromosomal Fragment Inversion Drives Fertility Restoration and Telomere Loss of C09 in Brassica oleracea","authors":"Wenjing Ren, Jinchao Si, Yiliao Feng, Hailong Yu, Jiamin Li, Fengqing Han, Xing Li, Limei Yang, Mu Zhuang, Honghao Lv, Jialei Ji, Feng Cheng, Yong Wang, Yangyong Zhang","doi":"10.1111/pbi.70396","DOIUrl":"https://doi.org/10.1111/pbi.70396","url":null,"abstract":"<p>Cabbage (<i>Brassica oleracea</i> L. var. <i>capitata</i>) is one of the most important vegetables grown globally (Li et al. <span>2024</span>). Currently, more than 90% of cabbage cultivars are male sterile. Male sterility-based hybridisation facilitates heterosis utilisation and yield enhancement (Han et al. <span>2023</span>). Ogura cytoplasmic male sterility (CMS) is more commonly used due to its maternal inheritance and easy transferability (Ren et al. <span>2020</span>, <span>2022</span>). However, CMS varieties cannot be self-pollinated, thereby limiting germplasm innovation and restricting genetic diversity, which can be solved by fertility restorer lines (Li et al. <span>2021</span>; Ren et al. <span>2022</span>). The <i>Rfo</i> gene exists only in radish but not in any <i>B. oleracea</i> crops. This gene has been transferred from radish into rapeseed, a related species of <i>B. oleracea</i>, providing a promising resource for creating cabbage Ogura CMS fertility restorer lines (FRLs) (Yu et al. <span>2020</span>). In our previous studies, <i>Rfo</i> was successfully transferred from rapeseed into Chinese kale and cabbage through distant hybridisation (Yu et al. <span>2020</span>; Ren et al. <span>2020</span>). However, critical knowledge gaps remain regarding the genome composition and genetic effects of the cabbage Ogura CMS restorer lines.</p>\u0000<p>Initially, the absence of <i>Rfo</i>-homozygous lines and high heterozygosity hindered <i>de novo</i> genome assembly of Ogura CMS FRLs (Table S1). In this study, BC<sub>7</sub> high-generation Ogura CMS FRLs were developed via successive backcrossing (Figure 1A). To overcome this challenge, we generated a chromosome-level genome assembly for the BC<sub>7</sub> FRL individual RFO7GH by combining Oxford Nanopore ultra-long reads, Illumina short reads for error correction and Hi-C scaffolding for chromosomal phasing (Figure S1A; Figure S2; Table S2). The <i>de novo</i> assembly yielded 40 contigs (N50 = 32.3 Mb) spanning 601.8 Mb from 59 Gb raw data. The final assembly contains more than 98.3% of the genome sequences (591.8 Mb) anchored to nine pseudochromosomes. Crucially, a 15.8-Mb alien radish genomic fragment (R09: 0–15 789 821 bp) containing the <i>Rfo</i> gene was inverted and integrated into the long-arm terminal region of C09 in RFO7GH, replacing the homologous 28.2-Mb cabbage genomic fragment (C09: 0–28 239 208 bp) (Figure 1B). Genome analysis revealed the loss of the telomere on the C09 long arm (Figure 1C). To validate this structural rearrangement, a specific primer set (GR-4F/4R) targeting the recombination breakpoint was designed, which successfully amplified a 4.6-kb junction sequence (Table S3). Sanger sequencing of the amplified products showed 100% concordance with Nanopore data, confirming the assembly accuracy.</p>\u0000<figure><picture>\u0000<source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/acaa9a00-9101-4b30-9f96-e267159a9b61/pbi70396-fig-0001-m.jpg\"/><img alt=\"Details are i","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"93 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651603","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}
Pablo Perez‐Colao, Gaetan Glauser, Jacobo Cruces, Jorge Lozano‐Juste, Manuel Rodriguez‐Concepcion
Plant biofortification with phytonutrients typically relies on metabolic engineering strategies known as ‘push’ (enhancing biosynthetic flux), ‘block’ (inhibiting competing pathways) and ‘pull’ (promoting metabolite storage). Here, we describe a novel synthetic compound, X57, that simultaneously targets biosynthesis, competition and storage to enhance leaf tocopherol content. Tocopherols protect plants against oxidative stress, have a dietary value as vitamin E and are highly appreciated antioxidants in food and cosmetic formulations. X57 exerts a primary ‘push’ effect by inducing tocopherol biosynthesis, in part by reactivating a direct pathway that reduces geranylgeranyl diphosphate (GGPP) to phytyl diphosphate, bypassing the need for chlorophyll‐derived phytol. Accordingly, X57 promotes tocopherol accumulation in etiolated seedlings and restores tocopherol synthesis in mutants deficient in phytol phosphorylation. The ‘block’ effect is mediated by down‐regulation of GGPP consumption for carotenoid synthesis. X57 also induces a ‘pull’ effect via proliferation of plastoglobules (PG), plastidial lipoprotein bodies that synthesise and store tocopherols. X57‐induced PG proliferation is driven by increased tocopherol levels and up‐regulation of genes for PG structural proteins such as fibrillins. The unveiled genetic networks simultaneously coordinating plastidial isoprenoid metabolism and plastid differentiation might only be present in higher plants, because X57 does not promote but reduces tocopherol accumulation in Marchantia polymorpha .
{"title":"A Chemical Probe for Increasing Leaf Tocopherol Levels by Coordinated Modulation of Biosynthesis, Competition and Storage","authors":"Pablo Perez‐Colao, Gaetan Glauser, Jacobo Cruces, Jorge Lozano‐Juste, Manuel Rodriguez‐Concepcion","doi":"10.1111/pbi.70459","DOIUrl":"https://doi.org/10.1111/pbi.70459","url":null,"abstract":"Plant biofortification with phytonutrients typically relies on metabolic engineering strategies known as ‘push’ (enhancing biosynthetic flux), ‘block’ (inhibiting competing pathways) and ‘pull’ (promoting metabolite storage). Here, we describe a novel synthetic compound, X57, that simultaneously targets biosynthesis, competition and storage to enhance leaf tocopherol content. Tocopherols protect plants against oxidative stress, have a dietary value as vitamin E and are highly appreciated antioxidants in food and cosmetic formulations. X57 exerts a primary ‘push’ effect by inducing tocopherol biosynthesis, in part by reactivating a direct pathway that reduces geranylgeranyl diphosphate (GGPP) to phytyl diphosphate, bypassing the need for chlorophyll‐derived phytol. Accordingly, X57 promotes tocopherol accumulation in etiolated seedlings and restores tocopherol synthesis in mutants deficient in phytol phosphorylation. The ‘block’ effect is mediated by down‐regulation of GGPP consumption for carotenoid synthesis. X57 also induces a ‘pull’ effect via proliferation of plastoglobules (PG), plastidial lipoprotein bodies that synthesise and store tocopherols. X57‐induced PG proliferation is driven by increased tocopherol levels and up‐regulation of genes for PG structural proteins such as fibrillins. The unveiled genetic networks simultaneously coordinating plastidial isoprenoid metabolism and plastid differentiation might only be present in higher plants, because X57 does not promote but reduces tocopherol accumulation in <jats:styled-content style=\"fixed-case\"> <jats:italic>Marchantia polymorpha</jats:italic> </jats:styled-content> .","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"55 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651185","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}
Anxiang Wang, Tian Meng, Ju Wang, Xiaomei Xu, Dan Li, Zhanqing Lv, Aiqi Yan, Mian Zhou, Bo Ding, Qiuying Yang
Tomato yellow leaf curl China virus (TYLCCNV) is a major agricultural pathogen and primary model for circular single‐stranded DNA (cssDNA) virus studies. The infectious clones of TYLCCNV and other cssDNA viruses are usually constructed as two tandem copies of the small viral genomes in Agrobacterium‐ mediated T‐DNA vectors. However, during our experiment, we observed that successive cultivation of the agrobacterial infectious clone of TYLCCNV led to a reduction or complete loss of its virulence in host plants. Further analysis revealed that the instability of this infectious clone is analogous to the mechanism of viral genome release from the dimeric infectious clones of cssDNA viruses during rolling circle replication, with key contributing factors being the activity of the viral Replication protein (Rep) and the replication origins. Unlike the infectious clones of RNA viruses, which often utilize introns to disrupt toxic protein coding sequences in order to achieve stabilization, the infectious clones of DNA viruses are unable to remove introns before releasing viral genomes. To address this challenge, we developed a micro‐homology mediated end joining (MMEJ)‐based system that disrupts the Rep coding sequences with an I‐ Sce I site flanked by microhomologous regions, stabilizing the infectious clone in prokaryotic cells. Then the transiently co‐expressed I‐ Sce I enzyme seamlessly removes the introduced I‐ Sce I site through MMEJ repair in plant hosts, resulting in efficient release of functional TYLCCNV viral genomes in planta . Theoretically, this approach can be applied to the construction of stable infectious clones for all plant DNA viruses.
{"title":"Stabilisation of Tomato Yellow Leaf Curl China Virus Infectious Clones Through Micro‐Homology Mediated End Joining","authors":"Anxiang Wang, Tian Meng, Ju Wang, Xiaomei Xu, Dan Li, Zhanqing Lv, Aiqi Yan, Mian Zhou, Bo Ding, Qiuying Yang","doi":"10.1111/pbi.70455","DOIUrl":"https://doi.org/10.1111/pbi.70455","url":null,"abstract":"<jats:italic>Tomato yellow leaf curl China virus</jats:italic> (TYLCCNV) is a major agricultural pathogen and primary model for circular single‐stranded DNA (cssDNA) virus studies. The infectious clones of TYLCCNV and other cssDNA viruses are usually constructed as two tandem copies of the small viral genomes in <jats:italic>Agrobacterium‐</jats:italic> mediated T‐DNA vectors. However, during our experiment, we observed that successive cultivation of the agrobacterial infectious clone of TYLCCNV led to a reduction or complete loss of its virulence in host plants. Further analysis revealed that the instability of this infectious clone is analogous to the mechanism of viral genome release from the dimeric infectious clones of cssDNA viruses during rolling circle replication, with key contributing factors being the activity of the viral Replication protein (Rep) and the replication origins. Unlike the infectious clones of RNA viruses, which often utilize introns to disrupt toxic protein coding sequences in order to achieve stabilization, the infectious clones of DNA viruses are unable to remove introns before releasing viral genomes. To address this challenge, we developed a micro‐homology mediated end joining (MMEJ)‐based system that disrupts the Rep coding sequences with an I‐ <jats:italic>Sce</jats:italic> I site flanked by microhomologous regions, stabilizing the infectious clone in prokaryotic cells. Then the transiently co‐expressed I‐ <jats:italic>Sce</jats:italic> I enzyme seamlessly removes the introduced I‐ <jats:italic>Sce</jats:italic> I site through MMEJ repair in plant hosts, resulting in efficient release of functional TYLCCNV viral genomes <jats:italic>in planta</jats:italic> . Theoretically, this approach can be applied to the construction of stable infectious clones for all plant DNA viruses.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"124 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145619427","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}
Wheat stripe rust (Puccinia striiformis f. sp. tritici, Pst) poses a catastrophic threat to global food security. While MADS-box transcription factors regulate development and abiotic stress, their roles in plant-pathogen immunity remain enigmatic. In this study, we identified TaMADS2, a Pst-induced MADS-box gene, as a positive regulator of wheat resistance. Functional analyses demonstrated that TaMADS2 overexpression significantly enhanced Pst resistance, whereas its knockdown rendered wheat more susceptible. Further investigation revealed that TaMADS2 interacts with trichome birefringence-like protein 21 (TaTBL21) to activate glycerol kinase-like (TaGKL) expression. Silencing TaGKL in wild-type or TaMADS2-overexpressing lines compromised resistance, with elevated Pst biomass. Notably, dual silencing of TaMADS2 and TaGKL further heightened susceptibility, confirming their synergistic defence role. Mechanistically, the TaMADS2-TaTBL21 complex promotes wheat resistance to stripe rust disease by upregulating TaGKL expression, leading to the accumulation of the key defence metabolites salicylic acid (SA) and glycerol-3-phosphate (G3P). Our study unveils a novel TaMADS2-TaTBL21-TaGKL module that potentiates wheat resistance against stripe rust, offering strategic targets for breeding resistant wheat.
{"title":"The TaMADS2-TaTBL21 Module Enhances Wheat Resistance to Stripe Rust by Activating TaGKL-Mediated Immunity.","authors":"Shijia Zhao,Hao Chu,Yanan Lu,Jiaqi Chen,Xu Wang,Guangxiang Tian,Mingjing Zhang,Pengyu Song,Yanhui Zhang,Guihua Bai,Gangming Zhan,Zhensheng Kang,Wenming Zheng,ZhengQing Fu,Na Liu","doi":"10.1111/pbi.70476","DOIUrl":"https://doi.org/10.1111/pbi.70476","url":null,"abstract":"Wheat stripe rust (Puccinia striiformis f. sp. tritici, Pst) poses a catastrophic threat to global food security. While MADS-box transcription factors regulate development and abiotic stress, their roles in plant-pathogen immunity remain enigmatic. In this study, we identified TaMADS2, a Pst-induced MADS-box gene, as a positive regulator of wheat resistance. Functional analyses demonstrated that TaMADS2 overexpression significantly enhanced Pst resistance, whereas its knockdown rendered wheat more susceptible. Further investigation revealed that TaMADS2 interacts with trichome birefringence-like protein 21 (TaTBL21) to activate glycerol kinase-like (TaGKL) expression. Silencing TaGKL in wild-type or TaMADS2-overexpressing lines compromised resistance, with elevated Pst biomass. Notably, dual silencing of TaMADS2 and TaGKL further heightened susceptibility, confirming their synergistic defence role. Mechanistically, the TaMADS2-TaTBL21 complex promotes wheat resistance to stripe rust disease by upregulating TaGKL expression, leading to the accumulation of the key defence metabolites salicylic acid (SA) and glycerol-3-phosphate (G3P). Our study unveils a novel TaMADS2-TaTBL21-TaGKL module that potentiates wheat resistance against stripe rust, offering strategic targets for breeding resistant wheat.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"10 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145613202","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}
Siqin Wang,Jiying Li,Zhang Wang,Miao Yu,Chang Liu,Dongye Liu,Ruohan Wang
The high demand for energy during floral thermogenesis drives the synergistic operation of multiple energy substrates for the rapid temperature rise and successful reproduction of flowers. However, how thermogenic plants precisely regulate substrate supply and metabolic pathways within a short time to support large-scale energy and heat release remains a mystery. This study revealed the elaborate synergistic supply mechanism of multi-source substrates in Magnolia denudata during thermogenesis. Transcriptome analysis showed that genes related to the tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) were significantly upregulated during the thermogenic stage (S2). Mitochondrial feeding assays using isotopically labelled substrates revealed that during the thermogenic stage, both the amount of pyruvate imported via the mitochondrial pyruvate carrier (MPC) and NAD-malic enzyme (NAD-ME) increased, and their synergistic effect accelerated the metabolic flow of the TCA cycle. Targeted lipidomics analysis indicated that the content of 63.6% fatty acids in the fatty acid degradation pathway decreased, while the key enzyme genes involved in triacylglycerol lipase (TGL) and fatty acid β-oxidation pathways were highly expressed during the thermogenic stage. In addition, enhanced expression of genes related to alanine aminotransferase (AlaAT) and glutamate dehydrogenase (GDH) suggested that amino acid metabolism might provide additional substrates for thermogenesis. This study clarifies the synergistic energy supply of carbohydrate, fatty acid and amino acid metabolism during thermogenesis in M. denudata, providing new evidence for understanding the metabolic regulatory flexibility of floral thermogenesis in plants.
{"title":"Multi-Omics Combined With Mitochondrial Feeding Assays Reveal a Novel Energy Metabolism Strategy for Floral Thermogenesis in Magnolia Driven by Synergistic Supply of Multiple Substrates.","authors":"Siqin Wang,Jiying Li,Zhang Wang,Miao Yu,Chang Liu,Dongye Liu,Ruohan Wang","doi":"10.1111/pbi.70479","DOIUrl":"https://doi.org/10.1111/pbi.70479","url":null,"abstract":"The high demand for energy during floral thermogenesis drives the synergistic operation of multiple energy substrates for the rapid temperature rise and successful reproduction of flowers. However, how thermogenic plants precisely regulate substrate supply and metabolic pathways within a short time to support large-scale energy and heat release remains a mystery. This study revealed the elaborate synergistic supply mechanism of multi-source substrates in Magnolia denudata during thermogenesis. Transcriptome analysis showed that genes related to the tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) were significantly upregulated during the thermogenic stage (S2). Mitochondrial feeding assays using isotopically labelled substrates revealed that during the thermogenic stage, both the amount of pyruvate imported via the mitochondrial pyruvate carrier (MPC) and NAD-malic enzyme (NAD-ME) increased, and their synergistic effect accelerated the metabolic flow of the TCA cycle. Targeted lipidomics analysis indicated that the content of 63.6% fatty acids in the fatty acid degradation pathway decreased, while the key enzyme genes involved in triacylglycerol lipase (TGL) and fatty acid β-oxidation pathways were highly expressed during the thermogenic stage. In addition, enhanced expression of genes related to alanine aminotransferase (AlaAT) and glutamate dehydrogenase (GDH) suggested that amino acid metabolism might provide additional substrates for thermogenesis. This study clarifies the synergistic energy supply of carbohydrate, fatty acid and amino acid metabolism during thermogenesis in M. denudata, providing new evidence for understanding the metabolic regulatory flexibility of floral thermogenesis in plants.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"71 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145613203","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}
Zhiyang Han, Buquan Zhao, Ming Fan, Shuangxi Zhang, Hao Peng, Xi Li, Surong Wang, Weihong Huang, Xingyu Yao, Chunwu Yang, Ke Wang, Xingguo Ye, Huali Tang
Wheat is one of the globally important food crops, but its genetically modified application lags behind other staple crops due to its complex genome and low transformation efficiency. Normally, traditional transgenic and gene‐edited wheat requires multiple selfing generations for obtaining homozygous lines, which delayed the development of genetically modified wheat varieties. In this study, we crossed a haploid inducer line with purple embryos (HIPE) as a male parent with a wheat cultivar Fielder, and then chose the haploid immature embryos without purple colour to be transformed with the Agrobacterium cells carrying the expression vectors with GUS , RUBY , and Cas9 and sgRNAs for targeting TaWaxy for generating both transgenic and gene editing wheat plants. Consequently, 139 haploid transgenic wheat plants were obtained with transformation efficiencies ranging from 66.7% to 78.4% according to molecular analysis, histochemical staining assay and colour observation. After chromosome doubling treatment to the haploid transgenic plantlets, homozygous transgenic and gene‐edited wheat lines were produced in T 0 generation. Genetic analysis of T 1 generation plants confirmed the stable inheritance of the transgenes and edited alleles without segregation. Finally, we established, for the first time, an efficient genetic transformation system based on the use of wheat haploid immature embryos, enabling direct acquisition of homozygous transgenic and gene‐edited wheat in T 0 generation. This approach can significantly shorten the wheat transgenic and gene‐edited breeding cycle and will provide a novel strategy for the genetic improvement of other polyploid crops.
{"title":"Rapid Genetic Stabilisation of Transgenic and Edited Wheat Through Transformation of Immature Haploid Embryos","authors":"Zhiyang Han, Buquan Zhao, Ming Fan, Shuangxi Zhang, Hao Peng, Xi Li, Surong Wang, Weihong Huang, Xingyu Yao, Chunwu Yang, Ke Wang, Xingguo Ye, Huali Tang","doi":"10.1111/pbi.70477","DOIUrl":"https://doi.org/10.1111/pbi.70477","url":null,"abstract":"Wheat is one of the globally important food crops, but its genetically modified application lags behind other staple crops due to its complex genome and low transformation efficiency. Normally, traditional transgenic and gene‐edited wheat requires multiple selfing generations for obtaining homozygous lines, which delayed the development of genetically modified wheat varieties. In this study, we crossed a haploid inducer line with purple embryos (HIPE) as a male parent with a wheat cultivar Fielder, and then chose the haploid immature embryos without purple colour to be transformed with the <jats:italic>Agrobacterium</jats:italic> cells carrying the expression vectors with <jats:italic>GUS</jats:italic> , <jats:italic>RUBY</jats:italic> , and <jats:italic>Cas9</jats:italic> and <jats:italic>sgRNAs</jats:italic> for targeting <jats:italic>TaWaxy</jats:italic> for generating both transgenic and gene editing wheat plants. Consequently, 139 haploid transgenic wheat plants were obtained with transformation efficiencies ranging from 66.7% to 78.4% according to molecular analysis, histochemical staining assay and colour observation. After chromosome doubling treatment to the haploid transgenic plantlets, homozygous transgenic and gene‐edited wheat lines were produced in T <jats:sub>0</jats:sub> generation. Genetic analysis of T <jats:sub>1</jats:sub> generation plants confirmed the stable inheritance of the transgenes and edited alleles without segregation. Finally, we established, for the first time, an efficient genetic transformation system based on the use of wheat haploid immature embryos, enabling direct acquisition of homozygous transgenic and gene‐edited wheat in T <jats:sub>0</jats:sub> generation. This approach can significantly shorten the wheat transgenic and gene‐edited breeding cycle and will provide a novel strategy for the genetic improvement of other polyploid crops.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"36 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145610902","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}
Yuan Zhong, Su Chen, Bo Sun, Min Liu, Zhenying Shi, Xuexia Miao, Haichao Li
The incidence of pests and diseases seriously impacts rice production, and NLR genes play a crucial role in the regulation of immune signalling in rice. Here, we identified an NLR gene OsRGA3 that positively regulates rice resistance to brown planthopper (BPH) and rice blast disease (RBD). The mutant OsRGA3 D605V as an auto‐activated form of OsRGA3 can form a resistosome and exhibit Ca 2+ permeable channel activity, triggering a hypersensitive response (HR) and providing rice with enhanced resistance to BPH, RBD and bacterial leaf blight (BLB). Biochemistry experiments confirmed that a Raf‐like MAPKKK OsILA1 interacts with OsRGA3 by Y2H, LCA, Pull‐down and BLI assay. OsILA1 negatively regulates rice resistance to BPH, RBD and BLB. Further study discovered that OsILA1 inhibits HR triggered by OsRGA3 D605V through phosphorylation of Y15 and Y138 sites to avoid an excessive immune response of plants. This study discovered a new MAPK‐NLR module with triple bio‐stress resistance that can be self‐activated and then deactivated by phosphorylation.
{"title":"An Auto‐Activated NLR ‐Protein OsRGA3 D605V Confers Rice Triple Resistance and Deactivates Resistance After Phosphorylation by OsILA1","authors":"Yuan Zhong, Su Chen, Bo Sun, Min Liu, Zhenying Shi, Xuexia Miao, Haichao Li","doi":"10.1111/pbi.70471","DOIUrl":"https://doi.org/10.1111/pbi.70471","url":null,"abstract":"The incidence of pests and diseases seriously impacts rice production, and <jats:italic>NLR</jats:italic> genes play a crucial role in the regulation of immune signalling in rice. Here, we identified an <jats:italic>NLR</jats:italic> gene <jats:italic>OsRGA3</jats:italic> that positively regulates rice resistance to brown planthopper (BPH) and rice blast disease (RBD). The mutant OsRGA3 <jats:sup>D605V</jats:sup> as an auto‐activated form of OsRGA3 can form a resistosome and exhibit Ca <jats:sup>2+</jats:sup> permeable channel activity, triggering a hypersensitive response (HR) and providing rice with enhanced resistance to BPH, RBD and bacterial leaf blight (BLB). Biochemistry experiments confirmed that a Raf‐like MAPKKK OsILA1 interacts with OsRGA3 by Y2H, LCA, Pull‐down and BLI assay. OsILA1 negatively regulates rice resistance to BPH, RBD and BLB. Further study discovered that OsILA1 inhibits HR triggered by OsRGA3 <jats:sup>D605V</jats:sup> through phosphorylation of Y15 and Y138 sites to avoid an excessive immune response of plants. This study discovered a new MAPK‐NLR module with triple bio‐stress resistance that can be self‐activated and then deactivated by phosphorylation.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"7 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145609195","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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