Plant Biotechnology Journal最新文献

{"title":"A Fusion Between Common Bean Phaseolin and the N-Terminal Domain of Maize 16 kDa Gamma Zein: Clues for Designing Nutritionally Improved Seed Storage Proteins","authors":"Emma Costa,&nbsp;Lucrezia Luciani,&nbsp;Mahsa Taghipour Rahimi,&nbsp;Alessandro Vitale,&nbsp;Emanuela Pedrazzini","doi":"10.1111/pbi.70719","DOIUrl":"10.1111/pbi.70719","url":null,"abstract":"<p>Increasing the nutritional value of seed storage proteins (SSPs) has been largely hindered by limited knowledge of how the features of the various SSP domains determine high accumulation in protein storage vacuoles (PSV) or as endoplasmic reticulum (ER)-located insoluble protein bodies (PB). We had previously designed zeolin, a fusion between phaseolin (PHSL, the main bean SSP, a soluble trimeric protein located in PSV) and the N-terminal domain of 27 kDa γ-zein (27γz, a major maize PB prolamin). Zeolin has a more balanced essential amino acid composition than each parental sequence and forms insoluble homomeric PB very similar to natural maize heteropolymeric PB, indicating that the zein domain has a dominant effect over PHSL. We had also determined that the paralog 16 kDa γ-zein (16γz) remains partially soluble and forms unusual threads in the ER when ectopically expressed. We now produced zeolin2 (zeo2), by fusing PHSL to the N-terminal domain of 16γz. We show that only a minor proportion of zeo2 remains in the ER, without forming PBs or threads. Zeo2 mainly forms soluble trimers with biochemical characteristics very similar to PHSL trimers, and traffics to the vacuole via the Golgi complex. This indicates that the 16γz domain fails to significantly dominate over PHSL. In transgenic arabidopsis, zeo2 accumulates to markedly higher levels than phaseolin and only slightly lower levels than zeolin. Zeo2 is also more accessible than zeolin to proteases during the early stages of seed germination. These findings provide new information relevant for the design of modified, nutritionally enhanced storage proteins.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5408-5424"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70719","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148388575","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":"Production of Functional Trastuzumab in Phaeodactylum tricornutum: Advancing Microalgae as Next-Generation Sustainable Antibody Biofactories","authors":"Charlotte Toustou,&nbsp;Manon Salvetti,&nbsp;Magalie Bénard,&nbsp;Christelle Fécourt,&nbsp;Sébastien Calbo,&nbsp;Kylian Jouie,&nbsp;France Blanchard,&nbsp;Camille N'Zué,&nbsp;Chloé Placines,&nbsp;Thomas Badou,&nbsp;Nicolas Piton,&nbsp;Patrice Lerouge,&nbsp;Ludovic Galas,&nbsp;Muriel Bardor","doi":"10.1111/pbi.70710","DOIUrl":"10.1111/pbi.70710","url":null,"abstract":"&lt;p&gt;The global demand for monoclonal antibodies (mAbs) is expanding at an unprecedented pace driven by their central role for the treatment of cancers, infectious and autoimmune diseases as well as their use in diagnostics. Access to these mAbs remains drastically uneven worldwide. Fewer than 5% of people can globally benefit from mAb-based therapies, and nearly half of the world's population lacks access to these diagnostic tools (Fleming et al. &lt;span&gt;2021&lt;/span&gt;). A major factor underlying this disparity is the high production cost of mAbs, which are predominantly manufactured in mammalian cells using Chinese Hamster Ovary (CHO) cells. The Gates Foundation has underscored that alleviating this global health inequity will require dramatic reductions in production costs. Therefore, identifying alternative low-cost expression systems represents a major challenge.&lt;/p&gt;&lt;p&gt;Unicellular eukaryotic microalgae recently emerged as a promising platform for mAbs affordable biomanufacturing (Hempel et al. &lt;span&gt;2011&lt;/span&gt;, &lt;span&gt;2017&lt;/span&gt;; Hempel and Maier &lt;span&gt;2012&lt;/span&gt;; Vanier et al. &lt;span&gt;2015&lt;/span&gt;, &lt;span&gt;2018&lt;/span&gt;). Microalgae offer a unique combination of advantages: they grow on inexpensive media, are photoautotrophic and do not contain human pathogens (Banerjee and Ward &lt;span&gt;2022&lt;/span&gt;). They naturally secrete very low levels of proteins into the culture medium (Chuberre et al. &lt;span&gt;2022&lt;/span&gt;), simplifying mAbs downstream processing and quality control. Importantly, while reaching growth rates comparable to bacteria, microalgae possess the eukaryotic machinery required for protein &lt;i&gt;N&lt;/i&gt;-glycosylation, essential for proper antibody folding, assembly and functionality (Vanier et al. &lt;span&gt;2015&lt;/span&gt;). At industrial scale for the same productivity, the mAb production in microalgae will be 10 times cheaper than in CHO cells (personal data).&lt;/p&gt;&lt;p&gt;Trastuzumab is a clinically validated CHO-produced mAb, approved for the treatment of HER2-positive breast cancer. It binds the Human Epidermal Growth Factor Receptor 2 (HER2), which is overexpressed in aggressive breast and stomach tumours. The global market for HER2-targeted diagnostic tests was valued at USD 166 million in 2025 and is projected to exceed USD 387 million by 2037 (CAGR + 7%). These trends reflect a growing demand for HER2-directed antibodies. Thus, developing a cost-effective production platform for Trastuzumab could meet the demands.&lt;/p&gt;&lt;p&gt;Here, we report the production of functional Trastuzumab in the diatom &lt;i&gt;Phaeodactylum tricornutum&lt;/i&gt; and demonstrate that microalgae-derived Trastuzumab exhibits functional bioequivalence to the commercially available CHO cells-produced antibody, supporting its use for research and diagnostic applications.&lt;/p&gt;&lt;p&gt;Trastuzumab was expressed in &lt;i&gt;P. tricornutum&lt;/i&gt; and accumulated in the culture medium, from which it was purified by protein A affinity chromatography allowing recovery of several mg/L of purified antibody. SDS-PAGE analysis, un","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5405-5407"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70710","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148388536","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":"PmD479 is an Unutilized Gene for Powdery Mildew Resistance in Common Wheat","authors":"Heng Tang,&nbsp;Bo Lyu,&nbsp;Yang Yu,&nbsp;Liwen Wang,&nbsp;Xiaomei Liu,&nbsp;Xuan Zheng,&nbsp;Baiqiang Yan,&nbsp;Ziqi Li,&nbsp;Tingting Zhang,&nbsp;Xuefei Chen,&nbsp;Lin Wang,&nbsp;Jing Zhang,&nbsp;Qiaoling Luo,&nbsp;Xuancheng Hao,&nbsp;Xingfeng Li,&nbsp;Junhua Peng,&nbsp;Lynn Epstein,&nbsp;Fei Ni,&nbsp;Fei He,&nbsp;Jiajie Wu,&nbsp;Daolin Fu","doi":"10.1111/pbi.70704","DOIUrl":"10.1111/pbi.70704","url":null,"abstract":"&lt;p&gt;Powdery mildew, caused by &lt;i&gt;Blumeria graminis&lt;/i&gt; f. sp. &lt;i&gt;tritici&lt;/i&gt; (&lt;i&gt;Bgt&lt;/i&gt;), threatens global wheat production. Wild emmer wheat (&lt;i&gt;Triticum turgidum&lt;/i&gt; ssp. &lt;i&gt;dicoccoides&lt;/i&gt;, DIC) is a progenitor of common wheat (&lt;i&gt;T. aestivum&lt;/i&gt; L.) and harbours abundant powdery mildew (&lt;i&gt;Pm&lt;/i&gt;) resistance genes (Wulff and Liu &lt;span&gt;2025&lt;/span&gt;). Here, we mapped and cloned &lt;i&gt;PmD479&lt;/i&gt;, an unutilized &lt;i&gt;Pm&lt;/i&gt; gene on chromosome 2BL in the DIC accession ‘DIC479’ (Marais et al. &lt;span&gt;2005&lt;/span&gt;), and introduced &lt;i&gt;PmD479&lt;/i&gt; into common wheat. Given its broad-spectrum efficacy and current rarity in common wheat germplasm, &lt;i&gt;PmD479&lt;/i&gt; is now a valuable resource for diversifying powdery mildew resistance in wheat.&lt;/p&gt;&lt;p&gt;DIC479 seedlings were resistant to 11 Chinese &lt;i&gt;Bgt&lt;/i&gt; isolates (Table S1). In response to &lt;i&gt;Bgt&lt;/i&gt; isolate E09, DIC479 produced reactive oxygen species (ROS) and exhibited localized necrosis (Figure 1A). DIC479 and Langdon (LDN, a &lt;i&gt;Pm&lt;/i&gt;-susceptible durum wheat, &lt;i&gt;T. turgidum ssp. durum&lt;/i&gt;) were used to develop an F&lt;sub&gt;2&lt;/sub&gt; population, which showed a 3:1 segregation (157 resistant vs. 47 susceptible; &lt;i&gt;χ&lt;/i&gt;&lt;sup&gt;2&lt;/sup&gt; = 0.418, &lt;i&gt;p&lt;/i&gt; &gt; 0.05). Among 83 randomly selected F&lt;sub&gt;2:3&lt;/sub&gt; families, 23 showed homozygous resistance, 39 segregated for &lt;i&gt;Pm&lt;/i&gt; resistance and 21 showed homozygous susceptibility, fitting a 1:2:1 ratio (&lt;i&gt;χ&lt;/i&gt;&lt;sup&gt;2&lt;/sup&gt; = 0.397, &lt;i&gt;p&lt;/i&gt; &gt; 0.05) (Table S2). Thus, &lt;i&gt;PmD479&lt;/i&gt; acts as a single dominant gene for &lt;i&gt;Pm&lt;/i&gt; resistance.&lt;/p&gt;&lt;p&gt;Nine &lt;i&gt;Pm&lt;/i&gt;-susceptible F&lt;sub&gt;2&lt;/sub&gt; plants were genotyped by the 90 K iSelect SNP array identified nine &lt;i&gt;PmD479&lt;/i&gt;-linked SNPs on the long arm of the homoeologous group 2 chromosome in wheat (Table S3). Sixteen molecular markers (Table S4) were developed by integrating the linked SNPs and the genomic collinearity among rice, barley, &lt;i&gt;Aegilops tauschii&lt;/i&gt; and common wheat. Using 204 F&lt;sub&gt;2&lt;/sub&gt; plants, &lt;i&gt;PmD479&lt;/i&gt; was mapped to a 2.35 cM interval between &lt;i&gt;Xsdau169&lt;/i&gt; and &lt;i&gt;Xsdau149&lt;/i&gt; on distal 2BL (Figure 1A).&lt;/p&gt;&lt;p&gt;To fine map &lt;i&gt;PmD479&lt;/i&gt;, we sequenced DIC479 with PacBio HiFi, generated 8923 contigs with an N50 of 6.05 Mb, and developed 13 additional markers (Table S4). Using 124 homozygous recombinants from 3045 F&lt;sub&gt;2:3&lt;/sub&gt; plants, &lt;i&gt;PmD479&lt;/i&gt; was precisely mapped between &lt;i&gt;Xsdau186B&lt;/i&gt; and &lt;i&gt;Xsdau193&lt;/i&gt; (&lt; 0.1 cM; Figure 1A; Tables S4 and S5). Both &lt;i&gt;Xsdau186B&lt;/i&gt; and &lt;i&gt;Xsdau193&lt;/i&gt; reside on the ptg000752l contig (3.17 Mb), delimit a 90 kb region in DIC479, and correspond to 811.78–811.93 Mb (2BL) in the Chinese Spring reference genome v2.1 (Zhu et al. &lt;span&gt;2021&lt;/span&gt;) (Figure 1A). &lt;i&gt;NLR148&lt;/i&gt;, a homologue of &lt;i&gt;TraesCS2B03G1573100&lt;/i&gt;, is the only complete gene in contig ptg000752l (Figure 1A; Table S6). &lt;i&gt;NLR148&lt;/i&gt; was constitutively expressed at 0 to 24 h post infestation (hpi) and then significantly upregulated at 48 to 96 hpi (Figure 1A). &lt;i&gt;NLR148&lt;/i&gt; encodes for a typical resistance ","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5317-5319"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70704","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148305136","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":"GhTTLL12 Coordinates With Transcriptional Regulators GhMML3 and GhMYB86 to Orchestrate Cotton Fibre Development Through Modulating Microtubule Dynamics","authors":"Xiaowei Ma,&nbsp;Xubinmiao Huang,&nbsp;Jinfang Li,&nbsp;Ting Zhao,&nbsp;Zhenfeng Ling,&nbsp;Shengcai Huang,&nbsp;Zesheng Rui,&nbsp;Yue Shi,&nbsp;Yulin Xing,&nbsp;Lei Sun,&nbsp;Mengting Wang,&nbsp;Haoxi Song,&nbsp;Rui Chen,&nbsp;Zequan Chen,&nbsp;Lei Shao,&nbsp;Nangan Sun,&nbsp;Junduo Wang,&nbsp;Juyun Zheng,&nbsp;Lei Fang","doi":"10.1111/pbi.70706","DOIUrl":"10.1111/pbi.70706","url":null,"abstract":"<p>Microtubules (MTs) are crucial for cell division, growth, development and morphogenesis in plants. Cotton fibres are single-celled trichomes that originate from the epidermal cells of the ovule, making them an excellent model for studying plant cell differentiation and rapid elongation. However, the roles of MTs in cotton fibre development remain incompletely understood. In this study, we identified <i>GhTTLL12</i>, a tubulin-tyrosine ligase-like protein 12, as a positive regulator of fibre initiation and elongation via <i>Gh</i>-<i>Gb</i> introgression analysis. <i>GhTTLL12</i> was preferentially expressed during the rapid elongation stage of cotton fibres. Overexpression of <i>GhTTLL12</i> enhanced plant height, root length, fibre cell protrusion number and fibre length in cotton. Conversely, CRISPR/Cas9-mediated knockout of <i>GhTTLL12</i> led to opposite phenotypes, thereby significantly reducing fibre quality. MT co-sedimentation and immunofluorescence assays demonstrated that GhTTLL12 binds directly to MTs and promotes their assembly while facilitating the formation of transverse MT arrays in elongating fibres. Further investigation of the molecular mechanisms revealed that after GhTTLL12 is recruited into the nucleus by GhTUB8, it promotes mitosis in ovule epidermal cells upon activation by GhMML3, increasing the number of fibre cell protrusions. GhMYB86, a negative regulator of cotton fibre elongation, represses <i>GhTTLL12</i> transcription in the nucleus, leading to attenuated mitotic activity. Cytoplasmic GhTTLL12 modulates fibre cell elongation by regulating MT assembly and ordered arrangement. Collectively, our findings define a GhMML3/GhMYB86-GhTTLL12-GhTUB8 regulatory module that links stage-specific transcriptional regulation to MT remodelling during cotton fibre development, providing new insights into the improvement of fibre quality and yield.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5320-5339"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70706","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148305134","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":"Metabolomic Analysis Reveals Structural Diversity of Pectic Polysaccharides From Prunus persica (L.) Batsch and Their Relationship With Purees Characteristics","authors":"Meng Liu,&nbsp;Xuan Liu,&nbsp;Shaojie Fu,&nbsp;Qianhui Yang,&nbsp;Christophe Blecker,&nbsp;Jinfeng Bi","doi":"10.1111/pbi.70709","DOIUrl":"10.1111/pbi.70709","url":null,"abstract":"<p>To clarify how pectin heterogeneity relates to puree quality, this study investigated associations among pectin fractions, metabolite profiles, and physicochemical properties of peach purees from 20 cultivars with distinct viscosity, turbidity, and colour. Metabolomic analysis showed that sugars, organic acids, and phenolic compounds were closely associated with the yield of chelator-soluble pectin (CSP), indicating coordinated variation between metabolite profiles and structurally integrated pectin fractions across cultivars. CSP yield was further related to resistance to deformation, as reflected by yield stress. In contrast, rheological behaviour after yielding was more strongly associated with water-soluble pectin (WSP), whose yield and structural features were linked to viscosity, viscoelastic balance, and flow behaviour. Phenolic compounds such as taxifolin and phlorizin chalcone were associated with puree colour. Collectively, these results establish an applied evaluation framework linking metabolite variation, pectin fraction distribution, and cultivar-dependent quality traits. By connecting micro-biochemical compositions (metabolite profiles and pectin fractions) with macroscopic processing-related properties, this framework provides insights into cell wall-related characteristics relevant to peach puree quality and processing performance.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5425-5439"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70709","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148400715","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":"Insights Into the Origin and Local Adaptation Evolution of the Cultivated Sesame With Telomere-to-Telomere High-Quality Genome","authors":"Weifei Yang,&nbsp;Hengchun Cao,&nbsp;Hui Guo,&nbsp;Hongmei Miao,&nbsp;Guiting Li,&nbsp;Yinghui Duan,&nbsp;Ming Ju,&nbsp;Qiuzhen Tian,&nbsp;Qin Ma,&nbsp;Pengjie Chang,&nbsp;Zhanyou Zhang,&nbsp;Cong Mu,&nbsp;Xiaoxu Feng,&nbsp;Libin Wei,&nbsp;Weixiu Hou,&nbsp;Andrew H. Paterson,&nbsp;Haiyang Zhang","doi":"10.1111/pbi.70714","DOIUrl":"10.1111/pbi.70714","url":null,"abstract":"<p>Sesame (<i>Sesamum indicum</i> L., 2<i>n</i> = 26) is one of the oldest oilseed crops and is often called the ‘queen of oilseeds’ due to its high content of unsaturated fatty acids and natural antioxidants. Despite its long history, the origin and global spread of cultivated sesame remain unresolved. We assembled a telomere-to-telomere (T2T), high-quality reference genome of sesame (cv. Yuzhi11) to investigate sequence differences between genomes and its origin and the local adaptation evolution of flowering time (DF). We generated a 305 Mb T2T sesame reference genome (cv. Yuzhi11) with &gt; 99.99% base-level accuracy, identifying 31 063 protein-coding genes. Repetitive elements accounted for 52.03% of the genome. Population genomic analysis of 927 accessions from 14 regions identified four major groups. Integrative analyses of linkage disequilibrium decay (LD), nucleotide diversity (π), and fixation index (<i>F</i>_ST) support East Africa as the center of origin, with subsequent migration through the Middle East, to South Asia, South-East Asia, East Asia and ultimately to other parts of the world. Genome-wide association studies (GWAS) and selection scans identified 30 genes associated with flowering time. <i>SiUBP16</i> is a candidate associated with 7.6% of DF variation. Early-flowering accessions carried up to 225 favourable alleles. A flowering time prediction model for high-latitude regions achieved 96% accuracy. We present a high-quality T2T reference genome for cultivated sesame, shedding light on its origin, evolutionary history, and regional flowering time adaptation. This genome insights valuable tools for breeding programs aimed at improving yield and environmental adaptation in sesame and related crops.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5372-5391"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70714","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148379893","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":"Prime Editing of Phytoene Synthase 1 in Rice for Seed Carotenoid Biofortification","authors":"Byeong-Hoon Kim,&nbsp;Yeo Jin Lee,&nbsp;Yong Jin Choi,&nbsp;Sooyeon Lim,&nbsp;Young Jin Park,&nbsp;Jae Kwang Kim,&nbsp;Sun-Hwa Ha","doi":"10.1111/pbi.70697","DOIUrl":"10.1111/pbi.70697","url":null,"abstract":"&lt;p&gt;Carotenoids are vital for human health as provitamin A sources and antioxidants, yet their levels remain low in staple crops. The development of ‘Golden Rice’ set a milestone in carotenoid biofortification through endosperm-specific expression of &lt;i&gt;PSY&lt;/i&gt; and &lt;i&gt;CrtI&lt;/i&gt;, with subsequent efforts focusing on PSY due to its rate-limiting role in carotenoid biosynthesis (Giuliano &lt;span&gt;2017&lt;/span&gt;; Zhou et al. &lt;span&gt;2022&lt;/span&gt;). However, CRISPR-Cas-based gain-of-function strategies targeting PSY remain limited in cereal crops. Here, we evaluated natural PSY variants for enhanced enzymatic activity and introduced a mutation into the &lt;i&gt;OsPSY1&lt;/i&gt; locus via prime editing, resulting in increased seed carotenoid accumulation with Golden Rice-like traits.&lt;/p&gt;&lt;p&gt;Previous genome editing efforts for gain-of-function modification of &lt;i&gt;PSY&lt;/i&gt; include adenine base editing of &lt;i&gt;ClPSY1&lt;/i&gt; introducing the K149E allele in watermelon, the suggested to affect substrate-binding affinity (Tian et al. &lt;span&gt;2026&lt;/span&gt;). To enhance PSY activity via genome editing, we selected two natural variants and one artificial mutant previously associated with increased carotenoid biosynthesis: MePSY2&lt;sup&gt;A191D&lt;/sup&gt; from a yellow-rooted cassava, ZmPSY1&lt;sup&gt;P257T&lt;/sup&gt; from predominantly yellow endosperm maize, and SlPSY1&lt;sup&gt;N136Y&lt;/sup&gt; from active tomato SlPSY2 (Welsch et al. &lt;span&gt;2010&lt;/span&gt;; Shumskaya et al. &lt;span&gt;2012&lt;/span&gt;; Cao et al. &lt;span&gt;2019&lt;/span&gt;). Finally, A184D and P267T were selected as candidate golden SNPs (Figure S1a), following protein sequence alignment that identified corresponding the residues in rice as OsPSY1&lt;sup&gt;A184&lt;/sup&gt;, OsPSY1&lt;sup&gt;P267&lt;/sup&gt;, and OsPSY1&lt;sup&gt;Y144&lt;/sup&gt; (Figure S1b).&lt;/p&gt;&lt;p&gt;To assess their effects on carotenoid biosynthesis, three &lt;i&gt;OsPSY1&lt;/i&gt; variants—&lt;i&gt;OsPSY1&lt;/i&gt;&lt;sup&gt;A184D&lt;/sup&gt;, &lt;i&gt;OsPSY1&lt;/i&gt;&lt;sup&gt;P267T&lt;/sup&gt;, and &lt;i&gt;OsPSY1&lt;/i&gt;&lt;sup&gt;A184D/P267T&lt;/sup&gt;—were generated by site-directed mutagenesis based on &lt;i&gt;OsPSY1&lt;/i&gt;&lt;sup&gt;WT&lt;/sup&gt; (Table S1). After removing the transit peptide (TP) predicted by UniProt (Table S2), &lt;i&gt;E. coli&lt;/i&gt; harbouring &lt;i&gt;pAC-85b&lt;/i&gt; (lacking &lt;i&gt;CrtB&lt;/i&gt;) was transformed with each truncated variant, with A184D-containing variants showing increased β-carotene production (~1.3-fold) and enhanced yellow pigmentation compared to the wild-type and P267T variant; this trend was recapitulated in rice calli expressing full-length variants, which exhibited ~4-fold higher β-carotene levels, &lt;i&gt;de novo&lt;/i&gt; α-carotene production, and distinct orange coloration (Figure 1a). Positioned near the Mg2+ binding D-rich motif (Figure S1), the A184D substitution did not alter the overall 3D structures of OsPSY1 but increased polar contacts with T181 from two to four (Figure 1b). Molecular dynamics simulations further indicated enhanced local stability under substrate-free conditions and stabilization of the active-site lid in the presence of GGPP, as evidenced by reduced RMSF (Figure S2b,d). Consistently, in vitro kinetic ass","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5245-5247"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13398656/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148203467","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":"A TAF10-ERF109 Transcriptional Module Directs Flavonoid-Based Stress Resilience and Yield Enhancement in Foxtail Millet and Wheat","authors":"Meng Zhang,&nbsp;Kangwei Wang,&nbsp;Jiayin Fan,&nbsp;Yue Zheng,&nbsp;Qi Zhang,&nbsp;Haoyang Du,&nbsp;Shenghui Xiao,&nbsp;Jinguang Huang,&nbsp;Kang Yan,&nbsp;Shizhong Zhang,&nbsp;Qiang He,&nbsp;Guanqing Jia,&nbsp;Chengchao Zheng,&nbsp;Xianmin Diao,&nbsp;Guodong Yang,&nbsp;Changai Wu","doi":"10.1111/pbi.70711","DOIUrl":"10.1111/pbi.70711","url":null,"abstract":"<p>To ensure sustainable agricultural production under escalating environmental constraints such as drought and salinity, innovative strategies are urgently needed. Here, we reveal in foxtail millet (<i>Setaria italica</i>) that transcription factors TAF10 and ERF109 form a functional complex which co-activates the expression of key flavonoid biosynthesis genes (<i>SiPAL</i>, <i>SiF3′H</i>, <i>SiFLS</i>, <i>SiF3H</i>), thereby enhancing abiotic stress tolerance. Application of specific flavonoids (apigenin, naringenin, hesperetin) rescues stress-sensitive phenotypes in <i>ERF109-RNAi</i> and <i>taf10</i> lines. Importantly, field-based application protocols for apigenin were established in both foxtail millet and wheat (<i>Triticum aestivum</i>), leading to yield increases of 12.3%–19.6% in wheat and 15.8%–23.9% in foxtail millet under drought and saline field conditions. Our study not only delineates a conserved transcriptional mechanism regulating flavonoid-mediated stress resilience but also delivers a translatable, eco-friendly biostimulant strategy to enhance crop productivity in stress-affected environments.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5352-5368"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70711","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148321703","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":"Assessing the Impact of Multigene Engineering on the Proteome: Omega-3 Camelina as a Case Study","authors":"Hongtao Zhang,&nbsp;Johnathan A. Napier,&nbsp;Frederica L. Theodoulou","doi":"10.1111/pbi.70712","DOIUrl":"10.1111/pbi.70712","url":null,"abstract":"&lt;p&gt;&lt;i&gt;Camelina sativa&lt;/i&gt; (Camelina) has emerged as a valuable platform for lipid metabolic engineering owing to its short life cycle, favourable seed oil profile and amenability to genetic transformation (Haslam et al. &lt;span&gt;2025&lt;/span&gt;). Previous studies have demonstrated the successful installation of non-native pathways for the biosynthesis of the omega-3 very-long-chain polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in seeds (Han et al. &lt;span&gt;2020&lt;/span&gt;, &lt;span&gt;2022&lt;/span&gt;; Figure S1). However, the broader consequences of introducing such a complex, multi-step pathway on endogenous processes remain largely uncharacterised.&lt;/p&gt;&lt;p&gt;In common with other lipid metabolic engineering studies, EPA and DHA production is associated with a reduction in seed oil content- the so-called oil yield penalty, which is poorly understood (Bates et al. &lt;span&gt;2014&lt;/span&gt;; Han et al. &lt;span&gt;2020&lt;/span&gt;). Moreover, studies in other engineered crops suggest that manipulating even a single gene can lead to substantial proteome reprogramming (Alvarez et al. &lt;span&gt;2015&lt;/span&gt;), highlighting the potential for both targeted and unintended effects on the proteome. Understanding these impacts is essential for evaluating the stability, performance and commercial viability of engineered crops. Quantifying transgene-encoded proteins is also important for understanding pathway performance.&lt;/p&gt;&lt;p&gt;Tandem mass tag (TMT)-based multiplexing and data independent acquisition (DIA)-based label-free quantification (LFQ) offer different advantages for profiling both endogenous and heterologous proteins: TMT workflows provide precise cross sample relative quantification, while DIA plus intensity-based absolute quantification (iBAQ) provides broader coverage and approximate absolute abundance estimates, allowing robust cross validation of differential abundance and pathway enrichment as well as stoichiometric insight. Here, we apply these complementary strategies to characterise the seed proteomes of elite EPA- and DHA-producing Camelina lines which differ in their oil yield penalty. DHA1 and EPA8 typically exhibit an oil yield penalty of 8 and 5 percentage points, respectively (Han et al. &lt;span&gt;2020&lt;/span&gt;, &lt;span&gt;2022&lt;/span&gt;; Figure S1). Our goals were to assess global proteomic changes associated with multigene engineering, to identify proteomic signatures associated with the differing oil yield penalties observed in EPA and DHA lines, and to quantify the relative abundance of transgene-encoded enzymes.&lt;/p&gt;&lt;p&gt;We profiled proteomes of wild type and engineered seeds (WT, DHA1 and EPA8 lines; Figure S1) at the mid-seed filling stage, 25 days after pollination, when the oil yield penalty is already established (Han et al. &lt;span&gt;2020&lt;/span&gt;). Gel electrophoresis revealed distinct protein profiles, with markedly reduced seed storage protein abundance in the engineered lines, particularly DHA1 (Figure 1a). TMTpro16-plex-based analysis (Figure S2) identif","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5480-5482"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/pbi.70712","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148429145","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":"Iterative Genome Engineering Platform Enables Efficient Sucrose Biosynthesis From CO2 in Photosynthetic Synechococcus elongatus UTEX 2973","authors":"Shubin Li,&nbsp;Tao Sun,&nbsp;Dailin Liu,&nbsp;Tong Zhang,&nbsp;Lei Chen,&nbsp;Weiwen Zhang","doi":"10.1111/pbi.70702","DOIUrl":"10.1111/pbi.70702","url":null,"abstract":"<p>The single crossover occurring via homologous recombination is a common phenomenon existing among microbes like <i>Escherichia coli</i>, <i>Bacillus subtilis</i>, <i>Vibrio natriegens</i>, <i>Gluconobacter oxydans</i> and most cyanobacteria species, threatening the stability of engineered strains and challenging iterative genetic engineering. Among them, we take the fast-growing cyanobacterium <i>Synechococcus elongatus</i> UTEX 2973 (Syn2973) as a representative study due to its promising roles for CO₂ fixation and bioconversion. We established three marker-free platforms to achieve stable genome recombination: (i) T4CROSS, which employs two plasmids and four rounds of single crossover; (ii) TRIPLEARM, which uses a single plasmid containing three homologous arms for three rounds of single crossover; and (iii) CRISPRARM, which integrates CRISPR/Cpf1-mediated genome editing with homologous recombination. As proof of concept, we employed the CRISPRARM platform for a three-step sequential engineering of the sucrose biosynthetic pathway. The final engineered strain produced 7.12 g L⁻¹ of sucrose within 4 days.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 10","pages":"5284-5299"},"PeriodicalIF":12.8,"publicationDate":"2026-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13398943/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148256539","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}