Diabetes Mellitus is an epidemic affecting > 500 million, claiming 6-7 million lives annually. Chemically synthesised Glucagon-like peptide-1 receptor agonists (GLP-1RAs) containing artificial amino acids reduce haemoglobin A1c and obesity but are not yet affordable and require invasive injections. High dosage requirement and gastrointestinal complications are among the current limitations of oral GLP-1RAs. Therefore, we expressed codon optimised Exenatide and Lixisenatide fused with Cholera-toxin B-subunit (CTB) in lettuce chloroplasts to facilitate their oral delivery, increase affordability, and patient compliance. Site-specific integration of transgene expression cassettes into the chloroplast genome and removal of the selectable marker gene from marker-free lettuce transplastomic lines were confirmed using three sets of PCR primers. Homoplasmy in transplastomic lines was confirmed in Southern blots by the absence of untransformed genomes. CTB-Exenatide and CTB-Lixisenatide expression levels were 1.94 and 3.64 mg/g plant powder in T0 generation and increased ~31 and ~48%, respectively in marker-removed T1 lines. Maternal inheritance of transgenes was confirmed by lack of segregation when seedlings were germinated in the selection medium before removal of the antibiotic resistance gene (aadA). Monosialotetrahexosylganglioside (GM1) ELISA confirmed pentameric assembly efficiency of both CTB-fusion proteins similar to commercial CTB standards. GLP-1 receptor binding confirmed functionality of CTB-Exenatide/CTB-Lixisenatide with statistical significance (***p < 0.001 by t-test) and post-translational amidation in chloroplasts. Expression of functional CTB-Exenatide and CTB-Lixisenatide in an edible marker-free system for the first time and much lower dosage requirement for functionality than recently developed synthetic GLP-1RAs paves the way for clinical studies to advance oral delivery of these affordable biologics.
{"title":"Engineering Marker-Free Lettuce Chloroplast Genome to Express Functional Glucagon-Like Peptide-1 Receptor Agonists Exenatide and Lixisenatide.","authors":"Rahul Singh,Henry Daniell","doi":"10.1111/pbi.70554","DOIUrl":"https://doi.org/10.1111/pbi.70554","url":null,"abstract":"Diabetes Mellitus is an epidemic affecting > 500 million, claiming 6-7 million lives annually. Chemically synthesised Glucagon-like peptide-1 receptor agonists (GLP-1RAs) containing artificial amino acids reduce haemoglobin A1c and obesity but are not yet affordable and require invasive injections. High dosage requirement and gastrointestinal complications are among the current limitations of oral GLP-1RAs. Therefore, we expressed codon optimised Exenatide and Lixisenatide fused with Cholera-toxin B-subunit (CTB) in lettuce chloroplasts to facilitate their oral delivery, increase affordability, and patient compliance. Site-specific integration of transgene expression cassettes into the chloroplast genome and removal of the selectable marker gene from marker-free lettuce transplastomic lines were confirmed using three sets of PCR primers. Homoplasmy in transplastomic lines was confirmed in Southern blots by the absence of untransformed genomes. CTB-Exenatide and CTB-Lixisenatide expression levels were 1.94 and 3.64 mg/g plant powder in T0 generation and increased ~31 and ~48%, respectively in marker-removed T1 lines. Maternal inheritance of transgenes was confirmed by lack of segregation when seedlings were germinated in the selection medium before removal of the antibiotic resistance gene (aadA). Monosialotetrahexosylganglioside (GM1) ELISA confirmed pentameric assembly efficiency of both CTB-fusion proteins similar to commercial CTB standards. GLP-1 receptor binding confirmed functionality of CTB-Exenatide/CTB-Lixisenatide with statistical significance (***p < 0.001 by t-test) and post-translational amidation in chloroplasts. Expression of functional CTB-Exenatide and CTB-Lixisenatide in an edible marker-free system for the first time and much lower dosage requirement for functionality than recently developed synthetic GLP-1RAs paves the way for clinical studies to advance oral delivery of these affordable biologics.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"3 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034066","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}
Upendra Kumari Wijesundara, Agnelo Furtado, Ardashir Kharabian Masouleh, Natalie L. Dillon, Heather E. Smyth, Robert J. Henry
Mango (Mangifera indica) is one of the most popular fruits cultivated in tropical and subtropical regions of the world. The availability of reference genomes helps to identify the genetic basis of important traits. Here, we report assembled high-quality chromosome-level genomes for the Australian mango cultivar ‘Kensington Pride’ and M.laurina, a wild relative, which shows resistance to anthracnose disease. PacBio HiFi sequencing with higher genome coverage enabled the assembly of both genomes with 100% completeness. Genome sizes of ‘Kensington Pride’ and M. laurina were 367 Mb and 379 Mb, respectively, with all 20 chromosomes in both genomes having telomeres at both ends. K-mer analysis revealed that these genomes are highly heterozygous and significant structural variations were identified between ‘Kensington Pride’, M. laurina, and the recently published genome of the cultivar ‘Irwin’. Functional annotation identified presence/absence variations of key genes involved in carotenoid, anthocyanin, and terpenoid biosynthesis, responsible for fruit colour and flavour in mango. Furthermore, the presence of a SNP in β-1,3-glucanase 2 gene, previously reported to be associated with anthracnose resistance, was analysed. Whole genome duplication analysis confirmed that mangoes have undergone two polyploidization events during their evolution. Analysis revealed a conserved pattern of colinear genes, although many colinear blocks were also identified on non-homologous chromosomes.
芒果（Mangifera indica）是世界热带和亚热带地区最受欢迎的水果之一。参考基因组的可用性有助于确定重要性状的遗传基础。在这里，我们报告了澳大利亚芒果品种‘Kensington Pride’和野生亲缘种M. laurina的高质量染色体水平基因组的组装，后者显示出对炭疽病的抗性。PacBio HiFi测序具有较高的基因组覆盖率，可以100%完成两个基因组的组装。“肯辛顿骄傲”和月牙花的基因组大小分别为367 Mb和379 Mb，两个基因组的所有20条染色体两端都有端粒。K-mer分析显示，这些基因组是高度杂合的，并且在‘ Kensington Pride ’， M. laurina和最近发表的栽培品种‘ Irwin ’的基因组之间发现了显着的结构差异。功能注释确定了参与类胡萝卜素、花青素和萜类生物合成的关键基因的存在/缺失变化，这些基因负责芒果的水果颜色和味道。此外，还分析了先前报道与炭疽病抗性相关的β-1,3-葡聚糖酶2基因中SNP的存在。全基因组重复分析证实芒果在进化过程中经历了两次多倍体化事件。分析揭示了共线基因的保守模式，尽管许多共线块也在非同源染色体上被鉴定出来。
{"title":"Chromosome-Scale Haplotype Genome Assemblies for the Australian Mango ‘Kensington Pride’ and a Wild Relative, Mangifera laurina, Provide Insights Into Anthracnose-Resistance and Volatile Compound Biosynthesis Genes","authors":"Upendra Kumari Wijesundara, Agnelo Furtado, Ardashir Kharabian Masouleh, Natalie L. Dillon, Heather E. Smyth, Robert J. Henry","doi":"10.1111/pbi.70556","DOIUrl":"https://doi.org/10.1111/pbi.70556","url":null,"abstract":"Mango (<i>Mangifera indica</i>) is one of the most popular fruits cultivated in tropical and subtropical regions of the world. The availability of reference genomes helps to identify the genetic basis of important traits. Here, we report assembled high-quality chromosome-level genomes for the Australian mango cultivar <b>‘</b>Kensington Pride<b>’</b> and <i>M.</i> <i>laurina</i>, a wild relative, which shows resistance to anthracnose disease. PacBio HiFi sequencing with higher genome coverage enabled the assembly of both genomes with 100% completeness. Genome sizes of <b>‘</b>Kensington Pride<b>’</b> and <i>M. laurina</i> were 367 Mb and 379 Mb, respectively, with all 20 chromosomes in both genomes having telomeres at both ends. K-mer analysis revealed that these genomes are highly heterozygous and significant structural variations were identified between <b>‘</b>Kensington Pride<b>’</b>, <i>M. laurina</i>, and the recently published genome of the cultivar <b>‘</b>Irwin<b>’</b>. Functional annotation identified presence/absence variations of key genes involved in carotenoid, anthocyanin, and terpenoid biosynthesis, responsible for fruit colour and flavour in mango. Furthermore, the presence of a SNP in β-1,3-glucanase 2 gene, previously reported to be associated with anthracnose resistance, was analysed. Whole genome duplication analysis confirmed that mangoes have undergone two polyploidization events during their evolution. Analysis revealed a conserved pattern of colinear genes, although many colinear blocks were also identified on non-homologous chromosomes.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"31 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146034147","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}
Jatropha curcas is a promising feedstock for biodiesel and bio-jet fuels production; however, its seed yield is constrained by limited inflorescences. SPL9 is a member of the SBP-box gene family that promotes the juvenile-to-adult phase transition. Accumulating evidence demonstrated that the miR156/SPL module plays important roles in regulating diverse plant developmental processes. Here, we reveal that JcSPL9 regulates both seed yield and oil content in Jatropha. JcSPL9 is highly expressed in fruits and upregulated with age in Jatropha. Overexpression of miR156-resistant JcSPL9 (rJcSPL9) significantly increased seed yield and oil content, whereas overexpression of JcmiR156a had the opposite effects. The highest seed yield in rJcSPL9 transgenic plants was 80.76% greater than that in the WT plants, with a concomitant 12.6% increase in seed oil content. Correspondingly, JcmiR156a transgenic plants displayed 51.67% lower seed yield and 8.28% lower seed oil content compared to WT. Additionally, seed oil fatty acid composition was significantly altered in both rJcSPL9 and JcmiR156a transgenic Jatropha and Arabidopsis, as well as in Arabidopsis spl9 mutants. The key oil biosynthesis genes, including JcWRI1, JcDGAT1, JcDGAT2, and JcOLEOSIN, were upregulated in rJcSPL9 transgenic seeds but downregulated in JcmiR156a transformants. This study provides the first evidence that the miR156/SPL9 module regulates lipid accumulation and fatty acid biosynthesis in seeds. These results highlight SPL9 as a promising target for enhancing oil yield and quality in Jatropha and other oilseed crops.
{"title":"SBP-Box Transcription Factor JcSPL9 Regulates Both Seed Yield and Oil Content in the Biofuel Plant Jatropha curcas.","authors":"Mingyong Tang,Xue Bai,Yaoping Xia,Ping Huang,Zeng-Fu Xu","doi":"10.1111/pbi.70558","DOIUrl":"https://doi.org/10.1111/pbi.70558","url":null,"abstract":"Jatropha curcas is a promising feedstock for biodiesel and bio-jet fuels production; however, its seed yield is constrained by limited inflorescences. SPL9 is a member of the SBP-box gene family that promotes the juvenile-to-adult phase transition. Accumulating evidence demonstrated that the miR156/SPL module plays important roles in regulating diverse plant developmental processes. Here, we reveal that JcSPL9 regulates both seed yield and oil content in Jatropha. JcSPL9 is highly expressed in fruits and upregulated with age in Jatropha. Overexpression of miR156-resistant JcSPL9 (rJcSPL9) significantly increased seed yield and oil content, whereas overexpression of JcmiR156a had the opposite effects. The highest seed yield in rJcSPL9 transgenic plants was 80.76% greater than that in the WT plants, with a concomitant 12.6% increase in seed oil content. Correspondingly, JcmiR156a transgenic plants displayed 51.67% lower seed yield and 8.28% lower seed oil content compared to WT. Additionally, seed oil fatty acid composition was significantly altered in both rJcSPL9 and JcmiR156a transgenic Jatropha and Arabidopsis, as well as in Arabidopsis spl9 mutants. The key oil biosynthesis genes, including JcWRI1, JcDGAT1, JcDGAT2, and JcOLEOSIN, were upregulated in rJcSPL9 transgenic seeds but downregulated in JcmiR156a transformants. This study provides the first evidence that the miR156/SPL9 module regulates lipid accumulation and fatty acid biosynthesis in seeds. These results highlight SPL9 as a promising target for enhancing oil yield and quality in Jatropha and other oilseed crops.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"142 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021634","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}
Fan Xiao,Chu-Kun Wang,Jiu-Cheng Zhang,Xin-Yue Jian,Ying Xiang,Wang-Jiang Zhang,Jin-Chao Meng,Wen-Yan Wang,Da-Gang Hu
The ripening of climacteric fruits is characterised by a sharp increase in ethylene production, coinciding with the conversion of starch into soluble sugars. However, the regulatory interplay between ethylene and starch degradation in apple remains largely unclear. Here, we report a negative correlation between starch accumulation and ethylene levels during late fruit development. Integrated transcriptomic analysis identified the α-amylase gene MdAMY1 as a key component of the ethylene-starch regulatory pathway. Functional characterisation confirmed that MdAMY1, an ethylene-responsive gene, acts as a positive regulator of starch-to-sugar conversion. Biochemical assays showed that the basic helix-loop-helix (bHLH) transcription factor MdbHLH149 directly represses MdAMY1 transcription. Furthermore, MdERF17-a negative regulator in ethylene signalling-interacts with MdbHLH149 and synergistically enhances this repression. A combination of GUS staining, quantitative enzyme activity assays and VIGS-based transient transformation demonstrated that the MdERF17-MdbHLH149-MdAMY1 module acts downstream of ethylene signalling to control starch degradation. Collectively, these findings establish that ethylene facilitates starch degradation by negatively regulating the MdERF17-MdbHLH149-MdAMY1 repression module.
{"title":"The MdERF17-MdbHLH149 Module Mediates Ethylene-Induced Starch Degradation Through the Transcriptional Repression of α-Amylase MdAMY1 in Apple.","authors":"Fan Xiao,Chu-Kun Wang,Jiu-Cheng Zhang,Xin-Yue Jian,Ying Xiang,Wang-Jiang Zhang,Jin-Chao Meng,Wen-Yan Wang,Da-Gang Hu","doi":"10.1111/pbi.70561","DOIUrl":"https://doi.org/10.1111/pbi.70561","url":null,"abstract":"The ripening of climacteric fruits is characterised by a sharp increase in ethylene production, coinciding with the conversion of starch into soluble sugars. However, the regulatory interplay between ethylene and starch degradation in apple remains largely unclear. Here, we report a negative correlation between starch accumulation and ethylene levels during late fruit development. Integrated transcriptomic analysis identified the α-amylase gene MdAMY1 as a key component of the ethylene-starch regulatory pathway. Functional characterisation confirmed that MdAMY1, an ethylene-responsive gene, acts as a positive regulator of starch-to-sugar conversion. Biochemical assays showed that the basic helix-loop-helix (bHLH) transcription factor MdbHLH149 directly represses MdAMY1 transcription. Furthermore, MdERF17-a negative regulator in ethylene signalling-interacts with MdbHLH149 and synergistically enhances this repression. A combination of GUS staining, quantitative enzyme activity assays and VIGS-based transient transformation demonstrated that the MdERF17-MdbHLH149-MdAMY1 module acts downstream of ethylene signalling to control starch degradation. Collectively, these findings establish that ethylene facilitates starch degradation by negatively regulating the MdERF17-MdbHLH149-MdAMY1 repression module.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"88 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021641","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}
Plant viruses drive widespread crop epidemics, yet the host plant responses across different cell types, particularly how these responses are influenced by cultivars with varying genetic backgrounds, including the presence of resistance (R) genes, remain poorly understood. Using tomato brown rugose fruit virus (ToBRFV) and two tomato cultivars, 'Jinpeng No. 1' (JP) and 'Rutgers' (RG), with different genetic backgrounds, this study used single-cell RNA sequencing to explore infection dynamics and responses at the cellular level. Results showed that ToBRFV accumulated to different levels in the two cultivars, likely due to differences in their genetic backgrounds, particularly the distinct genotypes of the Tm-22 and tm-2 alleles. Following infection, the composition of cell types in tomato leaves also varied between the two cultivars. While the entry or movement of ToBRFV in the JP cultivar was not fully prevented early on, the viral accumulation in certain cell types of this cultivar was restricted. ToBRFV alters signalling pathways based on cell type and cultivars. Pseudotime analysis revealed that, in JP plants, ToBRFV reverses expression of brassinosteroid (BR) positive regulators during mesophyll cell development. Silencing positive BR regulators increased infection in JP plants, while suppressing it in RG plants, linking BR signalling to JP-dependent resistance. Exogenous BR suppressed ToBRFV in JP but enhanced it in RG plants. This study reveals the differential involvement of BR signalling during viral infection in the two cultivars, offering a framework for future studies of plant-virus interactions.
{"title":"Dissecting the Cell-Type-Specific Response to an Emerging Tobamovirus in Tomato Reveals Cultivar-Dependent Involvement of Brassinosteroid Signalling.","authors":"Yuhong Zhang,Shan Bu,Yuxin Nie,Luyou Wang,Jiayi Liu,Junchen Xu,Jiejun Peng,Fei Yan,Jian Wu","doi":"10.1111/pbi.70559","DOIUrl":"https://doi.org/10.1111/pbi.70559","url":null,"abstract":"Plant viruses drive widespread crop epidemics, yet the host plant responses across different cell types, particularly how these responses are influenced by cultivars with varying genetic backgrounds, including the presence of resistance (R) genes, remain poorly understood. Using tomato brown rugose fruit virus (ToBRFV) and two tomato cultivars, 'Jinpeng No. 1' (JP) and 'Rutgers' (RG), with different genetic backgrounds, this study used single-cell RNA sequencing to explore infection dynamics and responses at the cellular level. Results showed that ToBRFV accumulated to different levels in the two cultivars, likely due to differences in their genetic backgrounds, particularly the distinct genotypes of the Tm-22 and tm-2 alleles. Following infection, the composition of cell types in tomato leaves also varied between the two cultivars. While the entry or movement of ToBRFV in the JP cultivar was not fully prevented early on, the viral accumulation in certain cell types of this cultivar was restricted. ToBRFV alters signalling pathways based on cell type and cultivars. Pseudotime analysis revealed that, in JP plants, ToBRFV reverses expression of brassinosteroid (BR) positive regulators during mesophyll cell development. Silencing positive BR regulators increased infection in JP plants, while suppressing it in RG plants, linking BR signalling to JP-dependent resistance. Exogenous BR suppressed ToBRFV in JP but enhanced it in RG plants. This study reveals the differential involvement of BR signalling during viral infection in the two cultivars, offering a framework for future studies of plant-virus interactions.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"276 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146005349","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}
Sea Island cotton (Gossypium barbadense) produces premium-quality fibres, yet the genetic basis underlying its fibre development remains elusive. Here, we identify two key non-synonymous single nucleotide polymorphisms (SNPs, G/C and G/A) in the gene Gbar_D13G024080, which encodes the TRANSMEMBRANE PROTEIN 209 (TMEM209). These SNPs resulted in amino acid changes (V/L and R/K), and are significantly correlated with the fibre length in Sea Island cotton. CRISPR-Cas9-mediated knockout of GbTMEM209 significantly enhanced fibre length and fibre strength in both G. hirsutum and G. barbadense. Conversely, overexpression of GbTMEM209 in G. hirsutum led to reduced fibre length. Further mechanistic investigation revealed that GbTMEM209 competitively interacts with GbHOX3 to impair its transcriptional activation on cell wall-loosening genes GbEXPA1 and GbRDL1. Moreover, during the elongation stage of the fibres, GbTMEM209 and GbHOX3 exhibit an antagonistic relationship, which jointly regulate the development of cotton fibres. Virus-induced gene silencing (VIGS) of GbHOX3, GbEXPA1, or GbRDL1 consistently resulted in shortened fibres in Sea Island cotton, validating their critical roles in fibre development. Our findings establish GbTMEM209 as a novel negative regulator of fibre elongation and uncover a protein competition-mediated transcriptional control mechanism in cotton fibre morphogenesis. These findings provide valuable genetic targets and conceptual insights for molecular breeding programs aimed at improving cotton fibre quality.
{"title":"Transmembrane Protein GbTMEM209 Inhibits Fibre Elongation via Competitive Interaction With GbHOX3 in Gossypium barbadense.","authors":"Kaiyun Jiang,Nan Zhao,Jie Kong,Anhui Guo,Yuqi Liu,Meng Wang,Zixin Zhou,Mengling Sun,Weiran Wang,Jiahui Zhu,Daojun Yuan,Zhuanxia Pan,Pengbo Li,Suen Liu,Baosheng Guo,Huijing Li,Jingrou Zhang,Bin Li,Zhanghao Xia,Beibei Lv,Junyi Geng,Baoliang Wang,Alifu Aierxi,Xianlong Zhang,Lili Tu,Jinping Hua","doi":"10.1111/pbi.70557","DOIUrl":"https://doi.org/10.1111/pbi.70557","url":null,"abstract":"Sea Island cotton (Gossypium barbadense) produces premium-quality fibres, yet the genetic basis underlying its fibre development remains elusive. Here, we identify two key non-synonymous single nucleotide polymorphisms (SNPs, G/C and G/A) in the gene Gbar_D13G024080, which encodes the TRANSMEMBRANE PROTEIN 209 (TMEM209). These SNPs resulted in amino acid changes (V/L and R/K), and are significantly correlated with the fibre length in Sea Island cotton. CRISPR-Cas9-mediated knockout of GbTMEM209 significantly enhanced fibre length and fibre strength in both G. hirsutum and G. barbadense. Conversely, overexpression of GbTMEM209 in G. hirsutum led to reduced fibre length. Further mechanistic investigation revealed that GbTMEM209 competitively interacts with GbHOX3 to impair its transcriptional activation on cell wall-loosening genes GbEXPA1 and GbRDL1. Moreover, during the elongation stage of the fibres, GbTMEM209 and GbHOX3 exhibit an antagonistic relationship, which jointly regulate the development of cotton fibres. Virus-induced gene silencing (VIGS) of GbHOX3, GbEXPA1, or GbRDL1 consistently resulted in shortened fibres in Sea Island cotton, validating their critical roles in fibre development. Our findings establish GbTMEM209 as a novel negative regulator of fibre elongation and uncover a protein competition-mediated transcriptional control mechanism in cotton fibre morphogenesis. These findings provide valuable genetic targets and conceptual insights for molecular breeding programs aimed at improving cotton fibre quality.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"65 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146005343","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}
Tongxiao Xu,Teng Wang,Cong Zhang,Yuan Cao,Xiaoyun He
Agricultural sustainability faces serious challenges from population growth, climate change and ecological degradation. Genetic modification (GM) technology can be regarded as a precise extension of the Green Revolution, aiming to balance yield enhancement with ecological integrity through biotechnology. To systematically examine global trend, this study conducts a bibliometric analysis using worldwide literature data from 1994 to 2024. The findings reveal a dual-core structure of international collaboration, centered on China and the United States. The United States is closely connected with Korea, Japan and the United Kingdom, forming a high-density cluster, while China engages with emerging regions in Southeast Asia and Africa through the Belt and Road Initiative. This initiative is intended to strengthen China's influence and is accompanied by the proliferation of technology in countries less endowed with resources. The technology lifecycle has been evolved through three distinct phases. Initially, the process of Agrobacterium-mediated transformation in tobacco plants was carried out, marking the beginning of transgenic development. This was followed by the implementation of RNA interference (RNAi) technology to silence multiple genes. Finally, a breakthrough happened through the development of CRISPR-Cas9 genome editing technologies. The analyses conducted in this study demonstrate the preponderance of CRISPR in contemporary research, thus suggesting that the industry places a premium on technological refinement. Hence, the future technological trajectory is predicted to focus on germplasm digitization, multi-gene editing, intelligent breeding and synthetic biology. Transgenic technology will serve as a foundational support for achieving sustainable food security in the forthcoming second green revolution.
{"title":"Bibliometric-Based Analysis of Global Trends and Collaborative Networks in Plant Genetic Engineering (1994-2024).","authors":"Tongxiao Xu,Teng Wang,Cong Zhang,Yuan Cao,Xiaoyun He","doi":"10.1111/pbi.70550","DOIUrl":"https://doi.org/10.1111/pbi.70550","url":null,"abstract":"Agricultural sustainability faces serious challenges from population growth, climate change and ecological degradation. Genetic modification (GM) technology can be regarded as a precise extension of the Green Revolution, aiming to balance yield enhancement with ecological integrity through biotechnology. To systematically examine global trend, this study conducts a bibliometric analysis using worldwide literature data from 1994 to 2024. The findings reveal a dual-core structure of international collaboration, centered on China and the United States. The United States is closely connected with Korea, Japan and the United Kingdom, forming a high-density cluster, while China engages with emerging regions in Southeast Asia and Africa through the Belt and Road Initiative. This initiative is intended to strengthen China's influence and is accompanied by the proliferation of technology in countries less endowed with resources. The technology lifecycle has been evolved through three distinct phases. Initially, the process of Agrobacterium-mediated transformation in tobacco plants was carried out, marking the beginning of transgenic development. This was followed by the implementation of RNA interference (RNAi) technology to silence multiple genes. Finally, a breakthrough happened through the development of CRISPR-Cas9 genome editing technologies. The analyses conducted in this study demonstrate the preponderance of CRISPR in contemporary research, thus suggesting that the industry places a premium on technological refinement. Hence, the future technological trajectory is predicted to focus on germplasm digitization, multi-gene editing, intelligent breeding and synthetic biology. Transgenic technology will serve as a foundational support for achieving sustainable food security in the forthcoming second green revolution.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"30 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145994961","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}
Reactive oxygen species (ROS) are signalling molecules that promote programmed cell death in animal and plant systems. However, their role in rice (Oryza sativa L.) anther development is unclear. In this study, we show that lower transcript levels of the metallothionein gene OsMT2b in japonica rice plants obtained by RNA interference (RNAi) resulted in a serious reduction in the seed setting rate. Observations of semi-thin sections of anthers indicated that tapetum degradation initiates early and ends late in OsMT2b-RNAi plants relative to the wild type (WT). Nitroblue tetrazolium staining and measurements of hydrogen peroxide contents showed that ROS contents are higher in OsMT2b-RNAi plants than in WT. Terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assays showed that abnormal programmed cell death in the tapetum results in sterile microspores. In addition, the OsMT2b-RNAi plants were sensitive to photoperiod; they were sterile under natural long-day conditions but almost fully fertile under natural short-day conditions, indicating that OsMT2b integrates photoperiod information into pollen development. The discovery of this rice material may enrich germplasm resources for two-line hybrid rice breeding, and further research may enable its application in two-line hybrid rice breeding.
{"title":"OsMT2b Regulates Pollen Development and ROS Homeostasis in a Photoperiod-Dependent Manner.","authors":"Ying He,Can Wang,Zilong Luo,Mingyang Ding,Yunyi Wen,Jiao Liu,Weiting Chen,Jing Li,Chuxiong Zhuang,Dagang Jiang","doi":"10.1111/pbi.70549","DOIUrl":"https://doi.org/10.1111/pbi.70549","url":null,"abstract":"Reactive oxygen species (ROS) are signalling molecules that promote programmed cell death in animal and plant systems. However, their role in rice (Oryza sativa L.) anther development is unclear. In this study, we show that lower transcript levels of the metallothionein gene OsMT2b in japonica rice plants obtained by RNA interference (RNAi) resulted in a serious reduction in the seed setting rate. Observations of semi-thin sections of anthers indicated that tapetum degradation initiates early and ends late in OsMT2b-RNAi plants relative to the wild type (WT). Nitroblue tetrazolium staining and measurements of hydrogen peroxide contents showed that ROS contents are higher in OsMT2b-RNAi plants than in WT. Terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assays showed that abnormal programmed cell death in the tapetum results in sterile microspores. In addition, the OsMT2b-RNAi plants were sensitive to photoperiod; they were sterile under natural long-day conditions but almost fully fertile under natural short-day conditions, indicating that OsMT2b integrates photoperiod information into pollen development. The discovery of this rice material may enrich germplasm resources for two-line hybrid rice breeding, and further research may enable its application in two-line hybrid rice breeding.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"22 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145994858","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}
Low temperature is a critical abiotic stress that imposes major constraints on the sustainable development of the fruit tree industry. Although exogenous dopamine has been shown to enhance cold tolerance in plants, its molecular mechanisms in apple ( Malus domestica ) remain poorly understood. In this study, we systematically investigated the role of dopamine in cold stress using exogenous dopamine application, overexpression (OE), and RNA interference (RNAi) of the MdTYDC (a key enzyme in dopamine biosynthesis). Our findings demonstrate that dopamine enhances cold resistance in apple through multiple mechanisms, including reducing reactive oxygen species accumulation, improving photosynthesis and stomatal function, promoting anthocyanin biosynthesis, and upregulating CBF genes. Molecular genetic analyses further revealed that MdICE1, a central transcriptional regulator, directly binds to cis‐regulatory elements in the MdTYDC promoter, thereby activating its transcription. Notably, we identified another bHLH transcription factor, MdFAMA, which interacts with MdICE1 and facilitates its binding to the MdTYDC promoter. This interaction amplifies dopamine biosynthesis and strengthens cold resistance. Moreover, exogenous dopamine treatment synergistically induced MdICE1 and MdFAMA expression, forming a positive feedback loop. This feedback mechanism establishes a hierarchical amplification of signalling, further reinforcing tolerance to low temperatures. Collectively, this study elucidates, for the first time, the molecular framework through which the MdICE1/MdFAMA ‐MdTYDC regulatory module orchestrates dopamine‐mediated cold tolerance in apple, providing novel insights into stress adaptation in perennial fruit crops.
{"title":"The MdICE1 / MdFAMA ‐ MdTYDC Transcriptional Module Confers Cold Tolerance by Regulating Dopamine Metabolism in Apple","authors":"Kexin Tan, Xinyang Song, Ziyi Xu, Hongzhen Zhu, Ying Zhang, Shuhan Xu, Zhijun Zhang, Pengmin Li, Fengwang Ma, Chao Li","doi":"10.1111/pbi.70544","DOIUrl":"https://doi.org/10.1111/pbi.70544","url":null,"abstract":"Low temperature is a critical abiotic stress that imposes major constraints on the sustainable development of the fruit tree industry. Although exogenous dopamine has been shown to enhance cold tolerance in plants, its molecular mechanisms in apple ( <jats:styled-content style=\"fixed-case\"> <jats:italic>Malus domestica</jats:italic> </jats:styled-content> ) remain poorly understood. In this study, we systematically investigated the role of dopamine in cold stress using exogenous dopamine application, overexpression (OE), and RNA interference (RNAi) of the <jats:italic>MdTYDC</jats:italic> (a key enzyme in dopamine biosynthesis). Our findings demonstrate that dopamine enhances cold resistance in apple through multiple mechanisms, including reducing reactive oxygen species accumulation, improving photosynthesis and stomatal function, promoting anthocyanin biosynthesis, and upregulating <jats:italic>CBF</jats:italic> genes. Molecular genetic analyses further revealed that MdICE1, a central transcriptional regulator, directly binds to cis‐regulatory elements in the <jats:italic>MdTYDC</jats:italic> promoter, thereby activating its transcription. Notably, we identified another bHLH transcription factor, MdFAMA, which interacts with MdICE1 and facilitates its binding to the <jats:italic>MdTYDC</jats:italic> promoter. This interaction amplifies dopamine biosynthesis and strengthens cold resistance. Moreover, exogenous dopamine treatment synergistically induced <jats:italic>MdICE1</jats:italic> and <jats:italic>MdFAMA</jats:italic> expression, forming a positive feedback loop. This feedback mechanism establishes a hierarchical amplification of signalling, further reinforcing tolerance to low temperatures. Collectively, this study elucidates, for the first time, the molecular framework through which the MdICE1/MdFAMA <jats:italic>‐MdTYDC</jats:italic> regulatory module orchestrates dopamine‐mediated cold tolerance in apple, providing novel insights into stress adaptation in perennial fruit crops.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"17 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145986492","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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