Sugar building blocks are crucial for the chemical diversity and biological activity of secondary metabolites. UDP-dependent glycosyltransferases (UGTs) play a pivotal role in the biosynthesis of glycosides in plants by catalysing the attachment of sugar moieties to various bioactive natural products. However, the biosynthesis of oligosaccharide-chain glycosides is often limited by the narrow substrate specificity of UGTs. In this study, we identify a regio-specific β-(1,6) glycosyltransferase, UGT94BY1, from Platycodon grandiflorum. UGT94BY1 exhibits broad substrate promiscuity and can transfer up to three sugar moieties to the C6-OH position of the glucosyl group in various triterpenoids and phenolic glycosides, thereby forming β-(1,6) oligoglucoside chains. To elucidate the mechanism underlying its substrate selectivity, we determined the crystal structure of the UGT94BY1 complex with UDP at a resolution of 2.0 Å. Molecular simulations revealed that a critical structural motif, comprising residues N84-M91, S141-L155 and R179-E186, plays a key role in recognizing sugar acceptors and facilitating chain elongation. Our study unveils a powerful glycosyltransferase for β-(1,6) oligoglucoside chain biosynthesis and highlights key regions involved in substrate recognition and sugar chain extension, providing valuable insights for designing UGTs with customized substrate specificities for biotechnological applications.
{"title":"Molecular characterization and structural basis of a promiscuous glycosyltransferase for β-(1,6) oligoglucoside chain glycosides biosynthesis","authors":"Zhennan Jiang, Nianhang Chen, Hao-Tian Wang, Yungang Tian, Xiaoyu Du, Ruibo Wu, Luqi Huang, Zi-Long Wang, Yuan Yuan","doi":"10.1111/pbi.70059","DOIUrl":"https://doi.org/10.1111/pbi.70059","url":null,"abstract":"Sugar building blocks are crucial for the chemical diversity and biological activity of secondary metabolites. UDP-dependent glycosyltransferases (UGTs) play a pivotal role in the biosynthesis of glycosides in plants by catalysing the attachment of sugar moieties to various bioactive natural products. However, the biosynthesis of oligosaccharide-chain glycosides is often limited by the narrow substrate specificity of UGTs. In this study, we identify a regio-specific <i>β</i>-(1,6) glycosyltransferase, UGT94BY1, from <i>Platycodon grandiflorum</i>. UGT94BY1 exhibits broad substrate promiscuity and can transfer up to three sugar moieties to the C6-OH position of the glucosyl group in various triterpenoids and phenolic glycosides, thereby forming <i>β</i>-(1,6) oligoglucoside chains. To elucidate the mechanism underlying its substrate selectivity, we determined the crystal structure of the UGT94BY1 complex with UDP at a resolution of 2.0 Å. Molecular simulations revealed that a critical structural motif, comprising residues N84-M91, S141-L155 and R179-E186, plays a key role in recognizing sugar acceptors and facilitating chain elongation. Our study unveils a powerful glycosyltransferase for <i>β</i>-(1,6) oligoglucoside chain biosynthesis and highlights key regions involved in substrate recognition and sugar chain extension, providing valuable insights for designing UGTs with customized substrate specificities for biotechnological applications.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"9 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143660378","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}
Pathogen infection in host plants can alter the attraction and adaptability of herbivorous insects. Female adult insects often exhibit selective behaviours based on their environmental experiences, enabling their offspring to avoid adverse conditions and ensuring healthy growth and development. However, comprehensive studies integrating both the perspectives of offspring fitness and host plant to validate the selective significance of such parental ‘Mother knows worst’ experiences remain limited. Building on our previous findings that female Conogethes punctiferalis (Yellow peach moth, YPM) adults exhibit oviposition avoidance behaviour towards corn infected with Trichoderma asperellum, we further confirmed that corn infected by T. asperellum significantly inhibits the growth and development of YPM larvae. Feeding on infected corn decreases larval gut microbiota diversity, core microbiota abundance and led to differential expression of key genes in juvenile hormone metabolic pathway. Moreover, the content of flavonoid wogonin, a secondary metabolite, was significantly increased in infected corn. In vitro feeding experiments revealed that wogonin negatively impacts YPM larval growth by causing the juvenile hormone accumulation and suppressing the abundance of core gut microbial strains. This study validates the adaptive significance of parental empiricism from the perspective of offspring, while further elucidating the mechanisms by which microbial-mediated plant resistance against insects, as well as for exploring and utilizing effective biocontrol resources against YPMs.
{"title":"Mother knows worst? Fungal infection enhances corn flavonoid of wogonin to inhibit Conogethes punctiferalis larval growth","authors":"Qian Li, Jiayu Li, Kaining Wu, Yue Tong, Aihuan Zhang, Yanli Du","doi":"10.1111/pbi.70051","DOIUrl":"https://doi.org/10.1111/pbi.70051","url":null,"abstract":"Pathogen infection in host plants can alter the attraction and adaptability of herbivorous insects. Female adult insects often exhibit selective behaviours based on their environmental experiences, enabling their offspring to avoid adverse conditions and ensuring healthy growth and development. However, comprehensive studies integrating both the perspectives of offspring fitness and host plant to validate the selective significance of such parental ‘Mother knows worst’ experiences remain limited. Building on our previous findings that female <i>Conogethes punctiferalis</i> (Yellow peach moth, YPM) adults exhibit oviposition avoidance behaviour towards corn infected with <i>Trichoderma asperellum</i>, we further confirmed that corn infected by <i>T. asperellum</i> significantly inhibits the growth and development of YPM larvae. Feeding on infected corn decreases larval gut microbiota diversity, core microbiota abundance and led to differential expression of key genes in juvenile hormone metabolic pathway. Moreover, the content of flavonoid wogonin, a secondary metabolite, was significantly increased in infected corn. In vitro feeding experiments revealed that wogonin negatively impacts YPM larval growth by causing the juvenile hormone accumulation and suppressing the abundance of core gut microbial strains. This study validates the adaptive significance of parental empiricism from the perspective of offspring, while further elucidating the mechanisms by which microbial-mediated plant resistance against insects, as well as for exploring and utilizing effective biocontrol resources against YPMs.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"33 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143653912","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}
Mingzhuo Li, Aditi Dey Poonam, Qirui Cui, Tzungfu Hsieh, Sumeetha Jagadeesan, Jin Xu, Wesley B. Bruce, Jonathan T. Vogel, Allen Sessions, Antonio Cabrera, Amanda C. Saville, Jean B. Ristaino, Rajesh Paul, Qingshan Wei
Crop breeding plays an essential role in addressing food security by enhancing crop yield, disease resistance and nutritional value. However, the current crop breeding process faces multiple challenges and limitations, especially in genotypic evaluations. Traditional methods for seed genotyping remain labour-intensive, time-consuming and cost-prohibitive outside of large-scale breeding programs. Here, we present a handheld microneedle (MN)-based seed DNA extraction platform for rapid, non-destructive and in-field DNA isolation from crop seeds for instant marker analysis. Using soybean seeds as a case study, we demonstrated the use of polyvinyl alcohol (PVA) MN patches for the successful extraction of DNA from softened soybean seeds. This extraction technology maintained high seed viability, showing germination rates of 82% and 79%, respectively, before and after MN sampling. The quality of MN-extracted DNA was sufficient for various genomic analyses, including PCR, LAMP and whole-genome sequencing. Importantly, this MN patch method also allowed for the identification of specific genetic differences between soybean varieties. Additionally, we designed a 3D-printed extraction device, which enabled multiplexed seed DNA extraction in a microplate format. In the future, this method could be applied at scale and in-field for crop seed DNA extraction and genotyping analysis.
作物育种通过提高作物产量、抗病性和营养价值，在解决粮食安全问题方面发挥着至关重要的作用。然而，当前的作物育种过程面临着多重挑战和限制，尤其是在基因型评估方面。传统的种子基因分型方法仍然是劳动密集型的，耗时耗力，在大规模育种计划之外成本高昂。在此，我们提出了一种基于手持微针（MN）的种子 DNA 提取平台，可快速、无损地在田间从作物种子中分离 DNA，用于即时标记分析。以大豆种子为例，我们展示了使用聚乙烯醇（PVA）微针贴片从软化的大豆种子中成功提取 DNA 的方法。这种提取技术保持了较高的种子活力，在 MN 取样前后的发芽率分别为 82% 和 79%。MN 提取的 DNA 质量足以进行各种基因组分析，包括 PCR、LAMP 和全基因组测序。重要的是，这种 MN 补丁方法还能识别大豆品种之间的特定遗传差异。此外，我们还设计了一种 3D 打印提取装置，可在微孔板格式下进行多重种子 DNA 提取。未来，这种方法可大规模应用于田间作物种子 DNA 提取和基因分型分析。
{"title":"Non-destructive seed genotyping via microneedle-based DNA extraction","authors":"Mingzhuo Li, Aditi Dey Poonam, Qirui Cui, Tzungfu Hsieh, Sumeetha Jagadeesan, Jin Xu, Wesley B. Bruce, Jonathan T. Vogel, Allen Sessions, Antonio Cabrera, Amanda C. Saville, Jean B. Ristaino, Rajesh Paul, Qingshan Wei","doi":"10.1111/pbi.70055","DOIUrl":"https://doi.org/10.1111/pbi.70055","url":null,"abstract":"Crop breeding plays an essential role in addressing food security by enhancing crop yield, disease resistance and nutritional value. However, the current crop breeding process faces multiple challenges and limitations, especially in genotypic evaluations. Traditional methods for seed genotyping remain labour-intensive, time-consuming and cost-prohibitive outside of large-scale breeding programs. Here, we present a handheld microneedle (MN)-based seed DNA extraction platform for rapid, non-destructive and in-field DNA isolation from crop seeds for instant marker analysis. Using soybean seeds as a case study, we demonstrated the use of polyvinyl alcohol (PVA) MN patches for the successful extraction of DNA from softened soybean seeds. This extraction technology maintained high seed viability, showing germination rates of 82% and 79%, respectively, before and after MN sampling. The quality of MN-extracted DNA was sufficient for various genomic analyses, including PCR, LAMP and whole-genome sequencing. Importantly, this MN patch method also allowed for the identification of specific genetic differences between soybean varieties. Additionally, we designed a 3D-printed extraction device, which enabled multiplexed seed DNA extraction in a microplate format. In the future, this method could be applied at scale and in-field for crop seed DNA extraction and genotyping analysis.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"14 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143660379","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}
Bohong Su, Hong Li, Ke Zhang, Haiyang Li, Caiyun Fan, Meiling Zhong, Hui Zou, Rujie Li, Liyu Chen, Jing Bo Jin, Mingkun Huang, Baohui Liu, Fanjiang Kong, Zhihui Sun
SummaryFormation of root nodules is a unique hallmark of the symbiotic interaction between legume host plants and rhizobia and is governed by a complex regulatory framework that balances the appropriate orchestration of rhizobial infection and subsequent nodule organogenesis. In contrast to prominent model species such as Medicago truncatula and Lotus japonicus, research on symbiotic signal transduction in the staple‐crop soybean Glycine max remains relatively insufficient. Here, we identified a soybean mutant with ~25% additional root nodules over wild‐type, designated as increased number of nodules 1 (inn1). Through map‐based cloning, INN1 encodes the EARLY FLOWERING 3a (ELF3a) protein component of the soybean Evening Complex, together with LUX1 and LUX2. INN1 is co‐expressed with LUX1 and LUX2 in roots, and knockout of INN1 or knockdown of LUX1 and LUX2 enhances root nodulation. The function of INN1 in negatively regulating nodulation is genetically and biochemically dependent upon LUXs, as the INN1–LUX complex binds to the promoter of the downstream pro‐nodulation target ENOD40, repressing its expression. ELF3a/INN1's repression of root‐nodule formation extends beyond its established roles in diverse above‐ground developmental and physiological processes and offers a theoretical basis for enhancing the biological‐nitrogen fixation capacity of soybean.
{"title":"Evening complex component ELF3 interacts with LUX proteins to repress soybean root nodulation","authors":"Bohong Su, Hong Li, Ke Zhang, Haiyang Li, Caiyun Fan, Meiling Zhong, Hui Zou, Rujie Li, Liyu Chen, Jing Bo Jin, Mingkun Huang, Baohui Liu, Fanjiang Kong, Zhihui Sun","doi":"10.1111/pbi.70053","DOIUrl":"https://doi.org/10.1111/pbi.70053","url":null,"abstract":"SummaryFormation of root nodules is a unique hallmark of the symbiotic interaction between legume host plants and rhizobia and is governed by a complex regulatory framework that balances the appropriate orchestration of rhizobial infection and subsequent nodule organogenesis. In contrast to prominent model species such as <jats:italic>Medicago truncatula</jats:italic> and <jats:italic>Lotus japonicus</jats:italic>, research on symbiotic signal transduction in the staple‐crop soybean <jats:italic>Glycine max</jats:italic> remains relatively insufficient. Here, we identified a soybean mutant with ~25% additional root nodules over wild‐type, designated as <jats:italic>increased number of nodules 1</jats:italic> (<jats:italic>inn1</jats:italic>). Through map‐based cloning, <jats:italic>INN1</jats:italic> encodes the EARLY FLOWERING 3a (ELF3a) protein component of the soybean Evening Complex, together with LUX1 and LUX2. <jats:italic>INN1</jats:italic> is co‐expressed with <jats:italic>LUX1</jats:italic> and <jats:italic>LUX2</jats:italic> in roots, and knockout of <jats:italic>INN1</jats:italic> or knockdown of <jats:italic>LUX1</jats:italic> and <jats:italic>LUX2</jats:italic> enhances root nodulation. The function of INN1 in negatively regulating nodulation is genetically and biochemically dependent upon LUXs, as the INN1–LUX complex binds to the promoter of the downstream pro‐nodulation target <jats:italic>ENOD40</jats:italic>, repressing its expression. ELF3a/INN1's repression of root‐nodule formation extends beyond its established roles in diverse above‐ground developmental and physiological processes and offers a theoretical basis for enhancing the biological‐nitrogen fixation capacity of soybean.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"16 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143640769","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}
SummaryFLOWERING LOCUS T (FT), a multifunctional regulator in crops, modulates multiple key agronomic traits such as flowering time or heading date and plant height; however, its role in grain development regulation is unclear. Herein, through genome‐wide association studies (GWAS), we identified TaFT‐D1, which encodes a phosphatidylethanolamine‐binding protein (PEBP), as a candidate gene for grain weight in wheat. A one‐bp insertion/deletion (InDel) (G/‐) in the third exon of TaFT‐D1, resulting in different protein lengths, was significantly associated with grain weight. TaFT‐D1 knockout via the CRISPR‐Cas9 system reduced grain size and weight, and TaFT‐D1 increased grain size by promoting cell proliferation and starch synthesis. Transcriptome analysis revealed a significant decrease in the expression of cell cycle‐ and starch synthesis‐related genes, including TaNAC019‐3A, TaSWEET15‐like‐7B, TaCYCD4;1 and TaCYCD3;2, in the taft‐d1 knockout line. TaFT‐D1 interacted with the bZIP transcription factor TaFDL2, and the tafdl2 mutant presented relatively small grains, suggesting that TaFDL2 is a positive regulator of grain size. Moreover, TaFDL2 bound to the promoters of downstream cell cycle‐ and starch synthesis‐related genes, activating their expression, whereas TaFT‐D1 increased this activation via TaFDL2. Interaction assays demonstrated that TaFT‐D1, Ta14‐3‐3A and TaFDL2 formed a regulatory complex. Furthermore, the TaFT‐D1(G) allele was significantly correlated with greater thousand‐grain weight and earlier heading. This favourable allele has undergone strong positive selection during wheat breeding in China. Our findings provide novel insights into how TaFT‐D1 regulates grain weight and highlight its potential application for yield improvement in wheat.
{"title":"TaFT‐D1 positively regulates grain weight by acting as a coactivator of TaFDL2 in wheat","authors":"Yinhui Zhang, Haixia Liu, Yaojia Wang, Xuemei Si, Yuxue Pan, Mengjiao Guo, Meijuan Wu, Yuanhao Li, Hongxia Liu, Xueyong Zhang, Jian Hou, Tian Li, Chenyang Hao","doi":"10.1111/pbi.70032","DOIUrl":"https://doi.org/10.1111/pbi.70032","url":null,"abstract":"Summary<jats:italic>FLOWERING LOCUS T</jats:italic> (<jats:italic>FT</jats:italic>), a multifunctional regulator in crops, modulates multiple key agronomic traits such as flowering time or heading date and plant height; however, its role in grain development regulation is unclear. Herein, through genome‐wide association studies (GWAS), we identified <jats:italic>TaFT‐D1</jats:italic>, which encodes a phosphatidylethanolamine‐binding protein (PEBP), as a candidate gene for grain weight in wheat. A one‐bp insertion/deletion (InDel) (G/‐) in the third exon of <jats:italic>TaFT‐D1</jats:italic>, resulting in different protein lengths, was significantly associated with grain weight. <jats:italic>TaFT‐D1</jats:italic> knockout via the CRISPR‐Cas9 system reduced grain size and weight, and <jats:italic>TaFT‐D1</jats:italic> increased grain size by promoting cell proliferation and starch synthesis. Transcriptome analysis revealed a significant decrease in the expression of cell cycle‐ and starch synthesis‐related genes, including <jats:italic>TaNAC019‐3A</jats:italic>, <jats:italic>TaSWEET15‐like‐7B</jats:italic>, <jats:italic>TaCYCD4;1</jats:italic> and <jats:italic>TaCYCD3;2</jats:italic>, in the <jats:italic>taft‐d1</jats:italic> knockout line. TaFT‐D1 interacted with the bZIP transcription factor TaFDL2, and the <jats:italic>tafdl2</jats:italic> mutant presented relatively small grains, suggesting that TaFDL2 is a positive regulator of grain size. Moreover, TaFDL2 bound to the promoters of downstream cell cycle‐ and starch synthesis‐related genes, activating their expression, whereas TaFT‐D1 increased this activation via TaFDL2. Interaction assays demonstrated that TaFT‐D1, Ta14‐3‐3A and TaFDL2 formed a regulatory complex. Furthermore, the <jats:italic>TaFT‐D1(G)</jats:italic> allele was significantly correlated with greater thousand‐grain weight and earlier heading. This favourable allele has undergone strong positive selection during wheat breeding in China. Our findings provide novel insights into how <jats:italic>TaFT‐D1</jats:italic> regulates grain weight and highlight its potential application for yield improvement in wheat.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"55 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143653344","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}
Flower scent is a crucial determiner in pollinator attraction and a significant horticultural trait in ornamental plants. Orchids, which have long been of interest in evolutionary biology and horticulture, exhibit remarkable diversity in floral scent type and intensity. However, the mechanisms underlying floral scent biosynthesis and regulation in orchids remain largely unexplored. In this study, we focus on floral scent in Cymbidium tracyanum, a wild species known for its strong floral fragrance and as a primary breeding parent of commercial Cymbidium hybrids. We present a chromosome-level genome assembly of C. tracyanum, totaling 3.79 Gb in size. Comparative genomic analyses reveal significant expansion of gene families associated with terpenoid biosynthesis and related metabolic pathways in C. tracyanum. Integrative analysis of genomic, volatolomic and transcriptomic data identified terpenoids as the predominant volatile components in the flowers of C. tracyanum. We characterized the spatiotemporal patterns of these volatiles and identified CtTPS genes responsible for volatile terpenoid biosynthesis, validating their catalytic functions in vitro. Dual-luciferase reporter assays, yeast one-hybrid assays and EMSA experiments confirmed that CtTPS2, CtTPS3, and CtTPS8 could be activated by various transcription factors (i.e., CtAP2/ERF1, CtbZIP1, CtMYB2, CtMYB3 and CtAP2/ERF4), thereby regulating the production of corresponding monoterpenes and sesquiterpenes. Our study elucidates the biosynthetic and regulatory mechanisms of floral scent in C. tracyanum, which is of great significance for the breeding of fragrant Cymbidium varieties and understanding the ecological adaptability of orchids. This study also highlights the importance of integrating multi-omics data in deciphering key horticultural traits in orchids.
{"title":"Integrative omics reveals mechanisms of biosynthesis and regulation of floral scent in Cymbidium tracyanum","authors":"Mengling Tu, Ningyawen Liu, Zheng-Shan He, Xiu-Mei Dong, Tian-Yang Gao, Andan Zhu, Jun-Bo Yang, Shi-Bao Zhang","doi":"10.1111/pbi.70025","DOIUrl":"https://doi.org/10.1111/pbi.70025","url":null,"abstract":"Flower scent is a crucial determiner in pollinator attraction and a significant horticultural trait in ornamental plants. Orchids, which have long been of interest in evolutionary biology and horticulture, exhibit remarkable diversity in floral scent type and intensity. However, the mechanisms underlying floral scent biosynthesis and regulation in orchids remain largely unexplored. In this study, we focus on floral scent in <i>Cymbidium tracyanum</i>, a wild species known for its strong floral fragrance and as a primary breeding parent of commercial <i>Cymbidium</i> hybrids. We present a chromosome-level genome assembly of <i>C. tracyanum</i>, totaling 3.79 Gb in size. Comparative genomic analyses reveal significant expansion of gene families associated with terpenoid biosynthesis and related metabolic pathways in <i>C. tracyanum</i>. Integrative analysis of genomic, volatolomic and transcriptomic data identified terpenoids as the predominant volatile components in the flowers of <i>C. tracyanum</i>. We characterized the spatiotemporal patterns of these volatiles and identified <i>CtTPS</i> genes responsible for volatile terpenoid biosynthesis, validating their catalytic functions <i>in vitro</i>. Dual-luciferase reporter assays, yeast one-hybrid assays and EMSA experiments confirmed that <i>CtTPS2</i>, <i>CtTPS3,</i> and <i>CtTPS8</i> could be activated by various transcription factors (i.e., CtAP2/ERF1, CtbZIP1, CtMYB2, CtMYB3 and CtAP2/ERF4), thereby regulating the production of corresponding monoterpenes and sesquiterpenes. Our study elucidates the biosynthetic and regulatory mechanisms of floral scent in <i>C. tracyanum</i>, which is of great significance for the breeding of fragrant <i>Cymbidium</i> varieties and understanding the ecological adaptability of orchids. This study also highlights the importance of integrating multi-omics data in deciphering key horticultural traits in orchids.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"24 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143635088","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}
Zhuonan Dai, Jiantao Guan, Han Miao, Diane M. Beckles, Xiaoping Liu, Xingfang Gu, Shaoyun Dong, Shengping Zhang
Vitamin A is a crucial yet scarce vitamin essential for maintaining normal metabolism and bodily functions in humans and can only be obtained from food. Carotenoids represent a diverse group of functional pigments that act as precursors for vitamins, hormones, aroma volatiles and antioxidants. As a vital vegetable in the world, elevated carotenoid levels in cucumber fruit produce yellow flesh, enhancing both visual appeal and nutritional value. However, the genetic mechanisms and regulatory networks governing yellow flesh in cucumbers remain inadequately characterized. In this study, we employed map-based cloning to identify a Carotenoid Cleavage Dioxygenase 1 (CsCCD1) as a key genetic factor influencing yellow flesh in cucumbers. A causal single nucleotide polymorphism (SNP) in the eighth intron of CsCCD1 led to aberrant splicing, resulting in a truncated transcript. The truncated protein has significantly decreased enzyme activity and increased carotenoid accumulation in the fruit. CRISPR/Cas9-generated CsCCD1 knockout mutants exhibited yellow flesh and significantly higher carotenoid content compared to wild-type cucumbers. Metabolic profiling indicated a marked accumulation of β-cryptoxanthin in the flesh of these knockout mutants. The intronic SNP was shown to perfectly segregate with yellow flesh in 159 diverse cucumber germplasms, particularly within the semi-wild ecotype Xishuangbanna, known for its substantial carotenoid accumulation. Furthermore, transient overexpression of CsCCD1 in yellow-fleshed Xishuangbanna cucumbers restored a white flesh phenotype, underscoring the critical role of CsCCD1 in determining flesh colour in both cultivated and semi-wild cucumbers. These findings lay a theoretical foundation for breeding high-nutrient yellow-fleshed cucumber varieties.
{"title":"An intronic SNP in the Carotenoid Cleavage Dioxygenase 1 (CsCCD1) controls yellow flesh formation in cucumber fruit (Cucumis sativus L.)","authors":"Zhuonan Dai, Jiantao Guan, Han Miao, Diane M. Beckles, Xiaoping Liu, Xingfang Gu, Shaoyun Dong, Shengping Zhang","doi":"10.1111/pbi.70034","DOIUrl":"https://doi.org/10.1111/pbi.70034","url":null,"abstract":"Vitamin A is a crucial yet scarce vitamin essential for maintaining normal metabolism and bodily functions in humans and can only be obtained from food. Carotenoids represent a diverse group of functional pigments that act as precursors for vitamins, hormones, aroma volatiles and antioxidants. As a vital vegetable in the world, elevated carotenoid levels in cucumber fruit produce yellow flesh, enhancing both visual appeal and nutritional value. However, the genetic mechanisms and regulatory networks governing yellow flesh in cucumbers remain inadequately characterized. In this study, we employed map-based cloning to identify a <i>Carotenoid Cleavage Dioxygenase 1</i> (<i>CsCCD1</i>) as a key genetic factor influencing yellow flesh in cucumbers. A causal single nucleotide polymorphism (SNP) in the eighth intron of <i>CsCCD1</i> led to aberrant splicing, resulting in a truncated transcript. The truncated protein has significantly decreased enzyme activity and increased carotenoid accumulation in the fruit. CRISPR/Cas9-generated <i>CsCCD1</i> knockout mutants exhibited yellow flesh and significantly higher carotenoid content compared to wild-type cucumbers. Metabolic profiling indicated a marked accumulation of β-cryptoxanthin in the flesh of these knockout mutants. The intronic SNP was shown to perfectly segregate with yellow flesh in 159 diverse cucumber germplasms, particularly within the semi-wild ecotype Xishuangbanna, known for its substantial carotenoid accumulation. Furthermore, transient overexpression of <i>CsCCD1</i> in yellow-fleshed Xishuangbanna cucumbers restored a white flesh phenotype, underscoring the critical role of <i>CsCCD1</i> in determining flesh colour in both cultivated and semi-wild cucumbers. These findings lay a theoretical foundation for breeding high-nutrient yellow-fleshed cucumber varieties.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"9 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143635085","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}
María Verónica Rodríguez, Diego Hernán Sánchez, Nicolás Glison, Cristian Damián Ríos, Patricia Verónica Demkura, Cristian Camilo Álvarez Correa, Luis Germán Fernández, Carla Valeria Filippi, Ruth Heinz, Pedro Pardo, Santiago Rentería, Lisandro Guillaumet, Roberto Luis Benech-Arnold
Grain sorghum (Sorghum bicolor L. moench) stands as a globally significant cereal crop but the adversity of pre-harvest sprouting (PHS) caused by reduced grain dormancy and moist conditions prior to harvest remains unsolved. Here, we identified a dormancy QTL using a Redlan×IS9530 RIL population, where parent lines are low in tannins and early flowering but otherwise contrasting in grain dormancy and plant height. We phenotyped this population in 2 years with informative PHS-related traits (grain germination index, embryo sensitivity to abscisic acid and in one year the actual natural sprouting), revealing a robust dormancy QTL in chromosome 9 (qDOR-9). This signal overlapped with associations found for plant height (caused by the dw1 locus, used for decades in sorghum improvement) and time to flowering. The effect of qDOR-9 was validated with independent near isogenic lines carrying the IS9530 “dormant” allele while maintaining the Redlan dw1 “short” allele. Additional analyses on Yellow Milo, from which the dw1 allele originated, implied that a low dormancy allele close to dw1 was introduced to Redlan—as well as to many other currently productive lines—by breeding efforts aimed at decreasing plant height, thus illustrating a new instance of genome erosion canalised by crop breeding. However, the introgression of qDOR-9 could enhance PHS tolerance in cultivated dw1-carrying backgrounds without affecting plant stature.
{"title":"Introgression of dwarfing allele dw1 reduced seed dormancy and increased pre-harvest sprouting susceptibility in grain sorghum converted lines","authors":"María Verónica Rodríguez, Diego Hernán Sánchez, Nicolás Glison, Cristian Damián Ríos, Patricia Verónica Demkura, Cristian Camilo Álvarez Correa, Luis Germán Fernández, Carla Valeria Filippi, Ruth Heinz, Pedro Pardo, Santiago Rentería, Lisandro Guillaumet, Roberto Luis Benech-Arnold","doi":"10.1111/pbi.70007","DOIUrl":"https://doi.org/10.1111/pbi.70007","url":null,"abstract":"Grain sorghum (<i>Sorghum bicolor</i> L. moench) stands as a globally significant cereal crop but the adversity of pre-harvest sprouting (PHS) caused by reduced grain dormancy and moist conditions prior to harvest remains unsolved. Here, we identified a dormancy QTL using a Redlan×IS9530 RIL population, where parent lines are low in tannins and early flowering but otherwise contrasting in grain dormancy and plant height. We phenotyped this population in 2 years with informative PHS-related traits (grain germination index, embryo sensitivity to abscisic acid and in one year the actual natural sprouting), revealing a robust dormancy QTL in chromosome 9 (qDOR-9). This signal overlapped with associations found for plant height (caused by the <i>dw1</i> locus, used for decades in sorghum improvement) and time to flowering. The effect of qDOR-9 was validated with independent near isogenic lines carrying the IS9530 “dormant” allele while maintaining the Redlan <i>dw1</i> “short” allele. Additional analyses on Yellow Milo, from which the <i>dw1</i> allele originated, implied that a low dormancy allele close to <i>dw1</i> was introduced to Redlan—as well as to many other currently productive lines—by breeding efforts aimed at decreasing plant height, thus illustrating a new instance of genome erosion canalised by crop breeding. However, the introgression of qDOR-9 could enhance PHS tolerance in cultivated <i>dw1</i>-carrying backgrounds without affecting plant stature.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"1 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143631251","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}
Erfeng Kou, Zhongxu Luo, Jingyi Ye, Xu Chen, Dan Lu, Markita P. Landry, Honglu Zhang, Huan Zhang
Global food production faces persistent threats from environmental challenges and pathogenic attacks, leading to significant yield losses. Conventional strategies to combat pathogens, such as fungicides and disease-resistant breeding, are limited by environmental contamination and emergence of pathogen resistance. Herein, we engineered sunlight-sensitive and biodegradable carbon dots (CDs) capable of generating reactive oxygen species (ROS), offering a novel and sustainable approach for plant protection. Our study demonstrates that CDs function as dual-purpose materials: priming plant immune responses and serving as broad-spectrum antifungal agents. Foliar application of CDs generated ROS under light, and the ROS could damage the plant cell wall and trigger cell wall-mediated immunity. Immune activation enhanced plant resistance against pathogens without compromising photosynthetic efficiency or yield. Specifically, spray treatment with CDs at 240 mg/L (2 mL per plant) reduced the incidence of grey mould in N. benthamiana and tomato leaves by 44% and 12%, respectively, and late blight in tomato leaves by 31%. Moreover, CDs (480 mg/L, 1 mL) combined with continuous sunlight irradiation (simulated by xenon lamp, 9.4 × 105 lux) showed a broad-spectrum antifungal activity. The inhibition ratios for mycelium growth were 66.5% for P. capsici, 8% for S. sclerotiorum and 100% for B. cinerea, respectively. Mechanistic studies revealed that CDs effectively inhibited mycelium growth by damaging hyphae and spore structures, thereby disrupting the propagation and vitality of pathogens. These findings suggest that CDs offer a promising, eco-friendly strategy for sustainable crop protection, with potential for practical agricultural applications that maintain crop yields and minimize environmental impact.
{"title":"Sunlight-sensitive carbon dots for plant immunity priming and pathogen defence","authors":"Erfeng Kou, Zhongxu Luo, Jingyi Ye, Xu Chen, Dan Lu, Markita P. Landry, Honglu Zhang, Huan Zhang","doi":"10.1111/pbi.70050","DOIUrl":"https://doi.org/10.1111/pbi.70050","url":null,"abstract":"Global food production faces persistent threats from environmental challenges and pathogenic attacks, leading to significant yield losses. Conventional strategies to combat pathogens, such as fungicides and disease-resistant breeding, are limited by environmental contamination and emergence of pathogen resistance. Herein, we engineered sunlight-sensitive and biodegradable carbon dots (CDs) capable of generating reactive oxygen species (ROS), offering a novel and sustainable approach for plant protection. Our study demonstrates that CDs function as dual-purpose materials: priming plant immune responses and serving as broad-spectrum antifungal agents. Foliar application of CDs generated ROS under light, and the ROS could damage the plant cell wall and trigger cell wall-mediated immunity. Immune activation enhanced plant resistance against pathogens without compromising photosynthetic efficiency or yield. Specifically, spray treatment with CDs at 240 mg/L (2 mL per plant) reduced the incidence of grey mould in <i>N. benthamiana</i> and tomato leaves by 44% and 12%, respectively, and late blight in tomato leaves by 31%. Moreover, CDs (480 mg/L, 1 mL) combined with continuous sunlight irradiation (simulated by xenon lamp, 9.4 × 10<sup>5</sup> lux) showed a broad-spectrum antifungal activity. The inhibition ratios for mycelium growth were 66.5% for <i>P. capsici</i>, 8% for <i>S. sclerotiorum</i> and 100% for <i>B. cinerea</i>, respectively. Mechanistic studies revealed that CDs effectively inhibited mycelium growth by damaging hyphae and spore structures, thereby disrupting the propagation and vitality of pathogens. These findings suggest that CDs offer a promising, eco-friendly strategy for sustainable crop protection, with potential for practical agricultural applications that maintain crop yields and minimize environmental impact.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"37 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143631373","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}
Taiyu Chen, Marta Hojka, Philip Davey, Yaqi Sun, Fei Zhou, Tracy Lawson, Peter J. Nixon, Yongjun Lin, Lu‐Ning Liu
SummaryAlthough Rubisco is the most abundant enzyme globally, it is inefficient for carbon fixation because of its low turnover rate and limited ability to distinguish CO2 and O2, especially under high O2 conditions. To address these limitations, phytoplankton, including cyanobacteria and algae, have evolved CO2‐concentrating mechanisms (CCM) that involve compartmentalizing Rubisco within specific structures, such as carboxysomes in cyanobacteria or pyrenoids in algae. Engineering plant chloroplasts to establish similar structures for compartmentalizing Rubisco has attracted increasing interest for improving photosynthesis and carbon assimilation in crop plants. Here, we present a method to effectively induce the condensation of endogenous Rubisco within tobacco (Nicotiana tabacum) chloroplasts by genetically fusing superfolder green fluorescent protein (sfGFP) to the tobacco Rubisco large subunit (RbcL). By leveraging the intrinsic oligomerization feature of sfGFP, we successfully created pyrenoid‐like Rubisco condensates that display dynamic, liquid‐like properties within chloroplasts without affecting Rubisco assembly and catalytic function. The transgenic tobacco plants demonstrated comparable autotrophic growth rates and full life cycles in ambient air relative to the wild‐type plants. Our study offers a promising strategy for modulating endogenous Rubisco assembly and spatial organization in plant chloroplasts via phase separation, which provides the foundation for generating synthetic organelle‐like structures for carbon fixation, such as carboxysomes and pyrenoids, to optimize photosynthetic efficiency.
{"title":"Engineering Rubisco condensation in chloroplasts to manipulate plant photosynthesis","authors":"Taiyu Chen, Marta Hojka, Philip Davey, Yaqi Sun, Fei Zhou, Tracy Lawson, Peter J. Nixon, Yongjun Lin, Lu‐Ning Liu","doi":"10.1111/pbi.70047","DOIUrl":"https://doi.org/10.1111/pbi.70047","url":null,"abstract":"SummaryAlthough Rubisco is the most abundant enzyme globally, it is inefficient for carbon fixation because of its low turnover rate and limited ability to distinguish CO<jats:sub>2</jats:sub> and O<jats:sub>2</jats:sub>, especially under high O<jats:sub>2</jats:sub> conditions. To address these limitations, phytoplankton, including cyanobacteria and algae, have evolved CO<jats:sub>2</jats:sub>‐concentrating mechanisms (CCM) that involve compartmentalizing Rubisco within specific structures, such as carboxysomes in cyanobacteria or pyrenoids in algae. Engineering plant chloroplasts to establish similar structures for compartmentalizing Rubisco has attracted increasing interest for improving photosynthesis and carbon assimilation in crop plants. Here, we present a method to effectively induce the condensation of endogenous Rubisco within tobacco (<jats:italic>Nicotiana tabacum</jats:italic>) chloroplasts by genetically fusing superfolder green fluorescent protein (sfGFP) to the tobacco Rubisco large subunit (RbcL). By leveraging the intrinsic oligomerization feature of sfGFP, we successfully created pyrenoid‐like Rubisco condensates that display dynamic, liquid‐like properties within chloroplasts without affecting Rubisco assembly and catalytic function. The transgenic tobacco plants demonstrated comparable autotrophic growth rates and full life cycles in ambient air relative to the wild‐type plants. Our study offers a promising strategy for modulating endogenous Rubisco assembly and spatial organization in plant chloroplasts via phase separation, which provides the foundation for generating synthetic organelle‐like structures for carbon fixation, such as carboxysomes and pyrenoids, to optimize photosynthetic efficiency.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"53 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143627535","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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