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Mycovirus Vector-Mediated RNAi for Effective Gene Knockdown in Pine Wood Nematodes. 松材线虫分枝病毒载体介导的RNAi基因敲除研究。
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-09 DOI: 10.1111/pbi.70567
Ruiling Bian, Yifan Zhang, Zhihao Zhang, Yuchi Bai, Peiqin Li, Guanghui Tang, Lihua Guo, Huan Liu, Ida Bagus Andika, Qiaoxia Shang, Liying Sun
{"title":"Mycovirus Vector-Mediated RNAi for Effective Gene Knockdown in Pine Wood Nematodes.","authors":"Ruiling Bian, Yifan Zhang, Zhihao Zhang, Yuchi Bai, Peiqin Li, Guanghui Tang, Lihua Guo, Huan Liu, Ida Bagus Andika, Qiaoxia Shang, Liying Sun","doi":"10.1111/pbi.70567","DOIUrl":"https://doi.org/10.1111/pbi.70567","url":null,"abstract":"","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146140481","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}
引用次数: 0
Synthetic Raphanobrassica Genome Reveals Functional and Evolutionary Insights Into Clubroot Resistance Genes on Chromosome R5. 合成油菜基因组揭示R5染色体上抗棍棒病基因的功能和进化见解。
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-09 DOI: 10.1111/pbi.70570
Xueqing Zhou, Meixiu Wu, Zhiquan Yang, Taihua Yang, Wenjing Li, Yingnan Liu, Yaru Shi, Aihua Wang, Peng Chen, Chao Liu, Qingyong Yang, Wenlin Yu, Chunyu Zhang

Clubroot, a severe soil-borne disease caused by Plasmodiophora brassicae, poses a severe threat to global production of Brassicaceae oilseed crops and vegetables. To date, there has been a serious lack of clubroot-resistant germplasms in Brassica napus (AACC), necessitating the urgent development of novel disease-resistant germplasm. We present a high-quality genome assembly of an artificially synthesised allotetraploid Raphanobrassica (RRCC), which exhibits broad-spectrum immunity to diverse P. brassicae pathotypes. Using a sesquidiploid RACC as a genetic bridge, we developed B. napus-R. sativus (AACC-R) monosomic addition lines and mapped a major clubroot resistance (CR) locus on chromosome R5. Comparative genomic analysis identified 30 candidate CR genes in this region. Notably, overexpression of CRR5.5.11, which encodes a receptor-like protein, via hairy root transformation conferred significant resistance in susceptible B. napus. Evolutionary and functional analyses revealed conserved homologues of CRR5.5.11 in diploid Isatis tinctoria and triploid turnip ECD04. Furthermore, the ECD04 allele CRA3.2.2 also conferred clubroot resistance, indicating that CRR5.5.11 and CRA3.2.2 have originated from the same diploid ancestor. Our study elucidates the genetic and evolutionary basis of clubroot resistance in Raphanobrassica, providing novel germplasm, gene resources and theoretical insights for clubroot-resistance breeding in rapeseed and other Brassicaceae crops.

{"title":"Synthetic Raphanobrassica Genome Reveals Functional and Evolutionary Insights Into Clubroot Resistance Genes on Chromosome R5.","authors":"Xueqing Zhou, Meixiu Wu, Zhiquan Yang, Taihua Yang, Wenjing Li, Yingnan Liu, Yaru Shi, Aihua Wang, Peng Chen, Chao Liu, Qingyong Yang, Wenlin Yu, Chunyu Zhang","doi":"10.1111/pbi.70570","DOIUrl":"https://doi.org/10.1111/pbi.70570","url":null,"abstract":"<p><p>Clubroot, a severe soil-borne disease caused by Plasmodiophora brassicae, poses a severe threat to global production of Brassicaceae oilseed crops and vegetables. To date, there has been a serious lack of clubroot-resistant germplasms in Brassica napus (AACC), necessitating the urgent development of novel disease-resistant germplasm. We present a high-quality genome assembly of an artificially synthesised allotetraploid Raphanobrassica (RRCC), which exhibits broad-spectrum immunity to diverse P. brassicae pathotypes. Using a sesquidiploid RACC as a genetic bridge, we developed B. napus-R. sativus (AACC-R) monosomic addition lines and mapped a major clubroot resistance (CR) locus on chromosome R5. Comparative genomic analysis identified 30 candidate CR genes in this region. Notably, overexpression of CRR5.5.11, which encodes a receptor-like protein, via hairy root transformation conferred significant resistance in susceptible B. napus. Evolutionary and functional analyses revealed conserved homologues of CRR5.5.11 in diploid Isatis tinctoria and triploid turnip ECD04. Furthermore, the ECD04 allele CRA3.2.2 also conferred clubroot resistance, indicating that CRR5.5.11 and CRA3.2.2 have originated from the same diploid ancestor. Our study elucidates the genetic and evolutionary basis of clubroot resistance in Raphanobrassica, providing novel germplasm, gene resources and theoretical insights for clubroot-resistance breeding in rapeseed and other Brassicaceae crops.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146140505","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}
引用次数: 0
CpERF-WRI1 Manipulates Ethylene Sensing by Regulating the Expression of CpERS1 and Fruit Ripening in Papaya. cperf - wr1通过调控木瓜CpERS1的表达和果实成熟调控乙烯感知。
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-09 DOI: 10.1111/pbi.70579
Ziling Wu, Jiahui Cai, Faiz Ur Rahman, Wang Zheng, Qiunan Zhu, Weixin Chen, Xueping Li, Xiaoyang Zhu

Ethylene plays a crucial role in fruit ripening and is perceived by specialised receptor proteins embedded in the endoplasmic reticulum (ER) membrane. As an ethylene antagonist, 1-methylcyclopropene (1-MCP) binds to these receptors and delays papaya ripening, but improper use can cause ripening disorders. The expression of CpERS1 and CpERF-WRI1 is markedly upregulated during ripening yet is suppressed by inappropriate 1-MCP treatments. Similar ripening disorders and expression patterns of CpERS1 and CpERF-WRI1 were observed in immature, chilled and heat-stressed papaya fruits, indicating that these genes may play key roles in ripening disorder. CpERF-WRI1 binds to and activates the CpERS1 promoter both in vitro and in vivo. Transient overexpression of CpERF-WRI1 in papaya and ectopic expression in tomato accelerated fruit ripening, increased CpERS1 expression and upregulated ripening-related pathways, whereas virus-induced gene silencing (VIGS) of CpERF-WRI1 in papaya delayed ripening and downregulated those pathways. Similarly, overexpressing CpERS1 in papaya and tomato accelerated ripening and boosted the expression of ripening-associated genes, while VIGS-mediated silencing of CpERS1 delayed ripening and suppressed these genes. Together, these results indicate that CpERF-WRI1 regulates CpERS1 expression and modulates ethylene sensing within the system II ethylene signalling pathway to control papaya ripening.

{"title":"CpERF-WRI1 Manipulates Ethylene Sensing by Regulating the Expression of CpERS1 and Fruit Ripening in Papaya.","authors":"Ziling Wu, Jiahui Cai, Faiz Ur Rahman, Wang Zheng, Qiunan Zhu, Weixin Chen, Xueping Li, Xiaoyang Zhu","doi":"10.1111/pbi.70579","DOIUrl":"https://doi.org/10.1111/pbi.70579","url":null,"abstract":"<p><p>Ethylene plays a crucial role in fruit ripening and is perceived by specialised receptor proteins embedded in the endoplasmic reticulum (ER) membrane. As an ethylene antagonist, 1-methylcyclopropene (1-MCP) binds to these receptors and delays papaya ripening, but improper use can cause ripening disorders. The expression of CpERS1 and CpERF-WRI1 is markedly upregulated during ripening yet is suppressed by inappropriate 1-MCP treatments. Similar ripening disorders and expression patterns of CpERS1 and CpERF-WRI1 were observed in immature, chilled and heat-stressed papaya fruits, indicating that these genes may play key roles in ripening disorder. CpERF-WRI1 binds to and activates the CpERS1 promoter both in vitro and in vivo. Transient overexpression of CpERF-WRI1 in papaya and ectopic expression in tomato accelerated fruit ripening, increased CpERS1 expression and upregulated ripening-related pathways, whereas virus-induced gene silencing (VIGS) of CpERF-WRI1 in papaya delayed ripening and downregulated those pathways. Similarly, overexpressing CpERS1 in papaya and tomato accelerated ripening and boosted the expression of ripening-associated genes, while VIGS-mediated silencing of CpERS1 delayed ripening and suppressed these genes. Together, these results indicate that CpERF-WRI1 regulates CpERS1 expression and modulates ethylene sensing within the system II ethylene signalling pathway to control papaya ripening.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146140558","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}
引用次数: 0
Knocking Out Two Polyphenol Oxidase Genes Significantly Improves Recombinant Protein Purification in Nicotiana benthamiana. 敲除两个多酚氧化酶基因显著提高了本菌烟重组蛋白的纯化。
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-08 DOI: 10.1111/pbi.70575
Hai-Ping Diao, Hui-Xin Meng, Xue-Jiao Xu, Zeng-Lin Zhang, Jian-Feng Zhang, Yong-Feng Guo, Inhwan Hwang, Shi-Jian Song

Efficient purification remains a key technical challenge affecting the recovery efficiency of recombinant proteins in plant-based systems. Complex metabolites, particularly polyphenols, often cause recombinant protein aggregation during purification. In this study, we identified two key polyphenol oxidase genes, PPOa and PPOb from Nicotiana benthamiana, as responsible for these effects. Using CRISPR-Cas9, we generated two ppoa;ppob double knockout lines that significantly improved the purification of virus surface proteins like SARS-CoV-2 Spike trimer and influenza HA trimer. These lines showed reduced polyphenol-protein interactions, minimised aggregation, and higher purification yields. Our work establishes a clean, high-efficiency N. benthamiana chassis for scalable recombinant protein production.

{"title":"Knocking Out Two Polyphenol Oxidase Genes Significantly Improves Recombinant Protein Purification in Nicotiana benthamiana.","authors":"Hai-Ping Diao, Hui-Xin Meng, Xue-Jiao Xu, Zeng-Lin Zhang, Jian-Feng Zhang, Yong-Feng Guo, Inhwan Hwang, Shi-Jian Song","doi":"10.1111/pbi.70575","DOIUrl":"https://doi.org/10.1111/pbi.70575","url":null,"abstract":"<p><p>Efficient purification remains a key technical challenge affecting the recovery efficiency of recombinant proteins in plant-based systems. Complex metabolites, particularly polyphenols, often cause recombinant protein aggregation during purification. In this study, we identified two key polyphenol oxidase genes, PPOa and PPOb from Nicotiana benthamiana, as responsible for these effects. Using CRISPR-Cas9, we generated two ppoa;ppob double knockout lines that significantly improved the purification of virus surface proteins like SARS-CoV-2 Spike trimer and influenza HA trimer. These lines showed reduced polyphenol-protein interactions, minimised aggregation, and higher purification yields. Our work establishes a clean, high-efficiency N. benthamiana chassis for scalable recombinant protein production.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146140566","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}
引用次数: 0
Ubiquitination-Mediated Degradation of SlVPS29 by the SlHSFA7-SlRNF185 Module Enhances Tomato Pollen Thermotolerance. SlHSFA7-SlRNF185模块介导的SlVPS29泛素化降解增强番茄花粉耐热性
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-06 DOI: 10.1111/pbi.70583
Xiaolin Geng, Qinqin Yang, Hongwei Li, Xunwen Peng, Wenbo Zhang, Huihui Wang, Yushan Li, Fang Li, Quanquan Chen, Tao Lin

High temperature has posed significant challenges to global agriculture, markedly leading to reduced fertility and yield losses in tomato (Solanum lycopersicum). Therefore, thermotolerance-conferring genes and loci are needed to further improve cultivated tomatoes. Here we identified an E3 ubiquitin ligase SlRNF185, containing a C3HC4-type RING-HC domain, that confers tomato pollen thermotolerance. As a predominant ubiquitination member, SlRNF185 could degrade the SlVPS29 protein induced by heat stress to enhance thermotolerance. Mechanistically, we found that a heat shock transcription factor SlHSFA7 dramatically activates the expression of SlRNF185 under heat stress and acts as a positive regulator of tomato pollen thermotolerance. Collectively, our findings reveal that a SlHSFA7-SlRNF185 genetic module regulates ubiquitination-mediated degradation of SlVPS29 under heat stress, providing the strategy for breeding thermotolerance tomato varieties.

{"title":"Ubiquitination-Mediated Degradation of SlVPS29 by the SlHSFA7-SlRNF185 Module Enhances Tomato Pollen Thermotolerance.","authors":"Xiaolin Geng, Qinqin Yang, Hongwei Li, Xunwen Peng, Wenbo Zhang, Huihui Wang, Yushan Li, Fang Li, Quanquan Chen, Tao Lin","doi":"10.1111/pbi.70583","DOIUrl":"https://doi.org/10.1111/pbi.70583","url":null,"abstract":"<p><p>High temperature has posed significant challenges to global agriculture, markedly leading to reduced fertility and yield losses in tomato (Solanum lycopersicum). Therefore, thermotolerance-conferring genes and loci are needed to further improve cultivated tomatoes. Here we identified an E3 ubiquitin ligase SlRNF185, containing a C3HC4-type RING-HC domain, that confers tomato pollen thermotolerance. As a predominant ubiquitination member, SlRNF185 could degrade the SlVPS29 protein induced by heat stress to enhance thermotolerance. Mechanistically, we found that a heat shock transcription factor SlHSFA7 dramatically activates the expression of SlRNF185 under heat stress and acts as a positive regulator of tomato pollen thermotolerance. Collectively, our findings reveal that a SlHSFA7-SlRNF185 genetic module regulates ubiquitination-mediated degradation of SlVPS29 under heat stress, providing the strategy for breeding thermotolerance tomato varieties.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146130433","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}
引用次数: 0
An Elite Haplotype of Nitrogen-Use-Efficiency Gene LHT5 Enhances Salt Tolerance in Rice. 氮素利用效率基因LHT5单倍型提高水稻耐盐性
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-06 DOI: 10.1111/pbi.70584
Saisai Wang, Xingzhou Jiang, Gaoming Chen, Wei Wu, Chen Xu, Mingyu Du, Shuji Xiang, Xinran Cheng, Yunlu Tian, Junjie Tan, Chunming Wang, Jianmin Wan

Amino acids serve as fundamental building blocks and signalling molecules in plants, orchestrating stress adaptation mechanisms against diverse biotic and abiotic environmental challenges. However, the mechanism by which plants alter their nutrient metabolism processes to coordinate nitrogen use efficiency (NUE) and salt tolerance remains elusive. Here, we identified a Lysine-Histidine-type transporter 5 (LHT5) gene through genome-wide association studies (GWAS) that enhances NUE via amino acid accumulation regulation. Further research showed that OsLHT5 also confers salt tolerance in rice by promoting proline biosynthesis through transcriptional upregulation of OsP5CS1 and OsP5CS2 genes, thereby increasing cellular proline levels for osmotic adjustment. Notably, we identified a functionally critical 30-bp deletion in the OsLHT5 coding region, designated as the elite haplotype LHT5HapA, which substantially enhances amino acid transport capacity and consequently improves both NUE and salt tolerance. Functional validation demonstrated that overexpression of LHT5HapA significantly increases amino acid content, nitrogen accumulation, grain yield and salt stress tolerance compared to the wildtype allele. This study establishes a novel molecular framework linking amino acid transport to the coordination of nutrient utilisation and stress tolerance, offering valuable genetic resources and breeding strategies for developing climate-resilient rice cultivars with enhanced productivity under both optimal and saline conditions.

{"title":"An Elite Haplotype of Nitrogen-Use-Efficiency Gene LHT5 Enhances Salt Tolerance in Rice.","authors":"Saisai Wang, Xingzhou Jiang, Gaoming Chen, Wei Wu, Chen Xu, Mingyu Du, Shuji Xiang, Xinran Cheng, Yunlu Tian, Junjie Tan, Chunming Wang, Jianmin Wan","doi":"10.1111/pbi.70584","DOIUrl":"https://doi.org/10.1111/pbi.70584","url":null,"abstract":"<p><p>Amino acids serve as fundamental building blocks and signalling molecules in plants, orchestrating stress adaptation mechanisms against diverse biotic and abiotic environmental challenges. However, the mechanism by which plants alter their nutrient metabolism processes to coordinate nitrogen use efficiency (NUE) and salt tolerance remains elusive. Here, we identified a Lysine-Histidine-type transporter 5 (LHT5) gene through genome-wide association studies (GWAS) that enhances NUE via amino acid accumulation regulation. Further research showed that OsLHT5 also confers salt tolerance in rice by promoting proline biosynthesis through transcriptional upregulation of OsP5CS1 and OsP5CS2 genes, thereby increasing cellular proline levels for osmotic adjustment. Notably, we identified a functionally critical 30-bp deletion in the OsLHT5 coding region, designated as the elite haplotype LHT5<sup>HapA</sup>, which substantially enhances amino acid transport capacity and consequently improves both NUE and salt tolerance. Functional validation demonstrated that overexpression of LHT5<sup>HapA</sup> significantly increases amino acid content, nitrogen accumulation, grain yield and salt stress tolerance compared to the wildtype allele. This study establishes a novel molecular framework linking amino acid transport to the coordination of nutrient utilisation and stress tolerance, offering valuable genetic resources and breeding strategies for developing climate-resilient rice cultivars with enhanced productivity under both optimal and saline conditions.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146130478","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}
引用次数: 0
Tuning Rice Gene Expression via In Situ Promoter Truncations Using a Prime Editing Library. 利用引体编辑文库原位截断启动子调控水稻基因表达。
IF 10.5 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-06 DOI: 10.1111/pbi.70587
Yuying Li, Birong Xu, Xuanchang Gao, Ying Wang, Xiaoshuang Liu, Rongfang Xu, Juan Li, Pengcheng Wei, Ruiying Qin

Promoter engineering holds immense potential for fine-tuning gene expression and optimising agronomic traits, yet conventional genome-editing tools face limitations in precision, scalability, and risk mitigation. Here, we develop Prime Editing-mediated Promoter Engineering (PEPE), a DSB-free platform integrating bidirectional Protospacer adjacent motif (PAM) recognition (NGG/CCN) with combinatorial duo-pegRNA strategies to achieve tiled, overlapping deletions across entire plant promoters. Applying PEPE to the 1.8-kb rice D53 promoter, we generated a mutant library with stepwise deletions. Edited alleles showed stable inheritance, and dual-method validation confirmed the precision at junctions. Quantitative profiling revealed functional modularity: core-region deletions suppressed D53 expression by 70%-85%, while a distal deletion (D53-G9C10) paradoxically upregulated transcription 2.2-fold, uncovering a cryptic repressor element. Phenotypic variation corresponded with transcriptional changes, establishing a direct link between cis-regulatory diversity and agronomic traits. By circumventing DSBs and enabling kilobase-scale CRE manipulation, PEPE establishes a robust framework for decoding promoter logic and accelerating trait pyramiding in crops. This study advances plant genome editing by merging precision with scalability, offering transformative potential for breeding climate-resilient, high-yield cultivars.

{"title":"Tuning Rice Gene Expression via In Situ Promoter Truncations Using a Prime Editing Library.","authors":"Yuying Li, Birong Xu, Xuanchang Gao, Ying Wang, Xiaoshuang Liu, Rongfang Xu, Juan Li, Pengcheng Wei, Ruiying Qin","doi":"10.1111/pbi.70587","DOIUrl":"https://doi.org/10.1111/pbi.70587","url":null,"abstract":"<p><p>Promoter engineering holds immense potential for fine-tuning gene expression and optimising agronomic traits, yet conventional genome-editing tools face limitations in precision, scalability, and risk mitigation. Here, we develop Prime Editing-mediated Promoter Engineering (PEPE), a DSB-free platform integrating bidirectional Protospacer adjacent motif (PAM) recognition (NGG/CCN) with combinatorial duo-pegRNA strategies to achieve tiled, overlapping deletions across entire plant promoters. Applying PEPE to the 1.8-kb rice D53 promoter, we generated a mutant library with stepwise deletions. Edited alleles showed stable inheritance, and dual-method validation confirmed the precision at junctions. Quantitative profiling revealed functional modularity: core-region deletions suppressed D53 expression by 70%-85%, while a distal deletion (D53-G9C10) paradoxically upregulated transcription 2.2-fold, uncovering a cryptic repressor element. Phenotypic variation corresponded with transcriptional changes, establishing a direct link between cis-regulatory diversity and agronomic traits. By circumventing DSBs and enabling kilobase-scale CRE manipulation, PEPE establishes a robust framework for decoding promoter logic and accelerating trait pyramiding in crops. This study advances plant genome editing by merging precision with scalability, offering transformative potential for breeding climate-resilient, high-yield cultivars.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.5,"publicationDate":"2026-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146130414","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}
引用次数: 0
GmRPS5 Promoter‐Driven CRISPR/LbCas12a Efficiently Generates Soybean Sextuple Mutants GmRPS5启动子驱动CRISPR/LbCas12a高效生成大豆六重突变体
IF 13.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-05 DOI: 10.1111/pbi.70588
Xiangchao Kong, Kexin Fan, Cuiping Xin, Minhui Lu, Yunlu Shi, Jie Jin, Qi‐Jun Chen
{"title":"GmRPS5 Promoter‐Driven CRISPR/LbCas12a Efficiently Generates Soybean Sextuple Mutants","authors":"Xiangchao Kong, Kexin Fan, Cuiping Xin, Minhui Lu, Yunlu Shi, Jie Jin, Qi‐Jun Chen","doi":"10.1111/pbi.70588","DOIUrl":"https://doi.org/10.1111/pbi.70588","url":null,"abstract":"","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"56 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146122080","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}
引用次数: 0
OsWRKY53‐OsGT1 Module Regulates Rice Tiller Development and Is Involved in Fine‐Tuning Strigolactone Signaling OsWRKY53‐OsGT1模块调控水稻分蘖发育并参与孤孤内酯信号的微调
IF 13.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-05 DOI: 10.1111/pbi.70578
Jiaqi Tang, Guilong Zhao, Jiangli Yang, Zhuo Chen, Zhipeng Hong, Xin Jin, Ziming Qiu, Zhenyu Wang, Xiufeng Li, Jijun Yan, Changhua Liu, Weiqiang Li, Jinfang Chu, Yuanhu Xuan, Xiaojie Tian, Qingyun Bu
Plant architecture, including plant height, tiller number, and leaf angle, is a critical determinant of rice yield. However, few genes have been identified that simultaneously regulate these traits and hold breeding value. We have previously shown that OsWRKY53 regulates the plant height and leaf angle via BR signalling. Here, we establish OsWRKY53 as a novel negative regulator of tillering in rice. The oswrky53 mutant exhibits a semi‐dwarf stature coupled with increased tiller number, representing a promising agronomic combination. Genetic and molecular analyses reveal that OsWRKY53 acts as a direct transcriptional activator of OsTB1 , thereby suppressing tiller formation. In addition, we found OsWRKY53 physically interacts with OsGT1, and their cooperative action synergistically enhances OsTB1 expression and suppresses tiller number. Intriguingly, the oswrky53 mutant exhibits reduced sensitivity to Strigolactone (SL) and increased SL contents. We further demonstrate that SL promotes degradation of OsWRKY53, and D53 interacts with and stabilises the OsWRKY53. Simultaneously, OsWRKY53 negatively regulates SL biosynthesis, enabling OsWRKY53 to function as a fine‐tuning regulator in the SL signalling pathway. Furthermore, OsGT1 exhibits subspecies‐specific regulation, with indica accessions carrying the OsGT1 581T allele showing significantly enhanced tillering capacity compared to japonica varieties. These findings collectively reveal the mechanism by which OsWRKY53 regulates the formation of tillers in rice, providing new genetic targets for semi‐dwarf and high‐tillering rice breeding.
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引用次数: 0
A Fusarium sacchari Glycoside Hydrolase 12 Protein FsEG1 Is a Major Virulence Factor During Sugarcane Infection and Confers Resistance to Pokkah Boeng Disease via the HIGS Strategy 镰刀菌糖苷水解酶12蛋白FsEG1是甘蔗感染过程中的一个主要毒力因子,并通过HIGS策略赋予甘蔗对Pokkah Boeng病的抗性
IF 13.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-02-04 DOI: 10.1111/pbi.70577
Deng Wu, Tianshu Hong, Lulu Wang, Qianqian Ren, Shichao Wang, Yixue Bao, Muqing Zhang, Wei Yao, Qin Hu
Pokkah Boeng disease (PBD), caused by Fusarium sacchari , has severely impacted the yield and quality of sugarcane, resulting in significant economic losses. However, the molecular interaction mechanisms between F. sacchari and sugarcane remain poorly understood. In this study, we identified the GH12 family protein FsEG1, secreted by F. sacchari , as a critical virulence factor. Further analysis demonstrates that the hydrolase activity of FsEG1 is essential for the full virulence of F. sacchari . The enhanced immune responses and cell death induced by FsEG1 in N. benthamiana depend on the recognition of oligosaccharide elicitors derived from the degradation of host cell walls by FsEG1, which are detected by membrane‐localised receptors NbWAKs and NbCERK1, and this process also necessitates RAR1 and MAP3Kα to facilitate intracellular signal transduction. Consequently, FsEG1 activates DTI (DAMP‐triggered immunity) rather than the conventional PTI. The stable transgenic sugarcane plants carrying the FsEG1 RNAi hairpin construct displayed high levels of resistance to F. sacchari and decreased FsEG1 expression, production of specific FsEG1 siRNA in transgenic HIGS sugarcane plants was confirmed by stem‐loop qRT‐PCR; at the same time, the stable transgenic sugarcane plants with ectopic expression of FsEG1 also showed enhanced PBD resistance with activated expression of defence‐related genes. Overall, these findings establish a foundational basis for investigating the molecular mechanisms that govern the interactions between F. sacchari and sugarcane and offering valuable insights into enhancing sugarcane's resistance to PBD.
甘蔗赤霉病(Pokkah Boeng disease, PBD)是由甘蔗镰刀菌(Fusarium sacchari)引起的一种病害,严重影响甘蔗产量和品质,造成重大经济损失。然而,糖酵母菌与甘蔗之间的分子相互作用机制尚不清楚。在这项研究中,我们发现由F. sacchari分泌的GH12家族蛋白FsEG1是一个关键的毒力因子。进一步分析表明,FsEG1的水解酶活性对糖酵母菌的完全毒力至关重要。benthamiana中FsEG1诱导的免疫应答增强和细胞死亡依赖于FsEG1对宿主细胞壁降解产生的寡糖激发子的识别,这些诱导子被膜定位受体NbWAKs和NbCERK1检测到,并且这一过程还需要RAR1和MAP3Kα来促进细胞内信号转导。因此,FsEG1激活DTI (DAMP触发的免疫)而不是传统的PTI。携带FsEG1 RNAi发夹构建体的稳定转基因甘蔗植株对F. sacchari表现出高水平的抗性,FsEG1表达降低,通过茎环qRT - PCR证实了转基因甘蔗植株中FsEG1特异性siRNA的产生;同时,异位表达FsEG1的稳定转基因甘蔗植株也表现出抗性增强,防御相关基因的表达被激活。综上所述,这些研究结果为研究糖精酵母与甘蔗相互作用的分子机制奠定了基础,并为提高甘蔗对PBD的抗性提供了有价值的见解。
{"title":"A Fusarium sacchari Glycoside Hydrolase 12 Protein FsEG1 Is a Major Virulence Factor During Sugarcane Infection and Confers Resistance to Pokkah Boeng Disease via the HIGS Strategy","authors":"Deng Wu, Tianshu Hong, Lulu Wang, Qianqian Ren, Shichao Wang, Yixue Bao, Muqing Zhang, Wei Yao, Qin Hu","doi":"10.1111/pbi.70577","DOIUrl":"https://doi.org/10.1111/pbi.70577","url":null,"abstract":"Pokkah Boeng disease (PBD), caused by <jats:italic>Fusarium sacchari</jats:italic> , has severely impacted the yield and quality of sugarcane, resulting in significant economic losses. However, the molecular interaction mechanisms between <jats:italic>F. sacchari</jats:italic> and sugarcane remain poorly understood. In this study, we identified the GH12 family protein FsEG1, secreted by <jats:italic>F. sacchari</jats:italic> , as a critical virulence factor. Further analysis demonstrates that the hydrolase activity of FsEG1 is essential for the full virulence of <jats:italic>F. sacchari</jats:italic> . The enhanced immune responses and cell death induced by FsEG1 in <jats:italic>N. benthamiana</jats:italic> depend on the recognition of oligosaccharide elicitors derived from the degradation of host cell walls by FsEG1, which are detected by membrane‐localised receptors NbWAKs and NbCERK1, and this process also necessitates RAR1 and MAP3Kα to facilitate intracellular signal transduction. Consequently, FsEG1 activates DTI (DAMP‐triggered immunity) rather than the conventional PTI. The stable transgenic sugarcane plants carrying the FsEG1 RNAi hairpin construct displayed high levels of resistance to <jats:italic>F. sacchari</jats:italic> and decreased FsEG1 expression, production of specific FsEG1 siRNA in transgenic HIGS sugarcane plants was confirmed by stem‐loop qRT‐PCR; at the same time, the stable transgenic sugarcane plants with ectopic expression of FsEG1 also showed enhanced PBD resistance with activated expression of defence‐related genes. Overall, these findings establish a foundational basis for investigating the molecular mechanisms that govern the interactions between <jats:italic>F. sacchari</jats:italic> and sugarcane and offering valuable insights into enhancing sugarcane's resistance to PBD.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"18 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2026-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146115587","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}
引用次数: 0
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Plant Biotechnology Journal
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