Tiancheng Xu, Yongmei Li, Xing Liu, Xue Yang, Zhejun Huang, Jianfeng Xing, Cuili Liang, Junyi Li, Yingchao Tan, Shengmin Zhang, Jiyan Qi, De Ye, Zhonghua Li, Jie Cao, Chaorong Tang, Kaiye Liu
Rubber particles (RPs) are specialized organelles for the biosynthesis and storage of natural rubber in rubber-producing plants. However, the mechanisms underlying the biogenesis and development of RPs remain unclear. In this study, two latex-specific cis-prenyltransferases (CPTs), TkCPT1 and TkCPT2, were identified in Taraxacum kok-saghyz, with almost identical orthologues retained across other Taraxacum species. For the first time, Tkcpt1 single and Tkcpt1/2 double mutants were successfully generated using the CRISPR/Cas9 system. Rubber biosynthesis was significantly depressed in Tkcpt1 mutants and completely blocked in Tkcpt1/2 mutants. The absence of RPs in the Tkcpt1/2 was confirmed using oil red O and Nile red staining, high-speed centrifugal stratification, cryo-SEM and TEM on fresh latex or laticifer cells. Transcriptomic and proteomic analyses revealed that, in the latex of Tkcpt1/2, rubber biosynthesis was blocked at the protein level, while metabolomic profiling indicated an enrichment of lipids and terpenoids. Furthermore, knockout of TkCPTL1, a latex-specific CPT-like gene that encodes a rubber transferase activator, resulted in outright disruption of rubber biosynthesis and RP ontogeny, a phenotype similar to that of Tkcpt1/2 mutants. These findings indicate that rubber biosynthesis is a driving force for the biogenesis and development of RPs, providing new insights into rubber production mechanisms.
{"title":"Rubber biosynthesis drives the biogenesis and development of rubber particles, the rubber-producing organelles","authors":"Tiancheng Xu, Yongmei Li, Xing Liu, Xue Yang, Zhejun Huang, Jianfeng Xing, Cuili Liang, Junyi Li, Yingchao Tan, Shengmin Zhang, Jiyan Qi, De Ye, Zhonghua Li, Jie Cao, Chaorong Tang, Kaiye Liu","doi":"10.1111/pbi.70052","DOIUrl":"https://doi.org/10.1111/pbi.70052","url":null,"abstract":"Rubber particles (RPs) are specialized organelles for the biosynthesis and storage of natural rubber in rubber-producing plants. However, the mechanisms underlying the biogenesis and development of RPs remain unclear. In this study, two latex-specific <i>cis</i>-prenyltransferases (CPTs), TkCPT1 and TkCPT2, were identified in <i>Taraxacum kok-saghyz</i>, with almost identical orthologues retained across other <i>Taraxacum</i> species. For the first time, <i>Tkcpt1</i> single and <i>Tkcpt1/2</i> double mutants were successfully generated using the CRISPR/Cas9 system. Rubber biosynthesis was significantly depressed in <i>Tkcpt1</i> mutants and completely blocked in <i>Tkcpt1/2</i> mutants. The absence of RPs in the <i>Tkcpt1/2</i> was confirmed using oil red O and Nile red staining, high-speed centrifugal stratification, cryo-SEM and TEM on fresh latex or laticifer cells. Transcriptomic and proteomic analyses revealed that, in the latex of <i>Tkcpt1/</i>2, rubber biosynthesis was blocked at the protein level, while metabolomic profiling indicated an enrichment of lipids and terpenoids. Furthermore, knockout of <i>TkCPTL1</i>, a latex-specific <i>CPT-like</i> gene that encodes a rubber transferase activator, resulted in outright disruption of rubber biosynthesis and RP ontogeny, a phenotype similar to that of <i>Tkcpt1/2</i> mutants. These findings indicate that rubber biosynthesis is a driving force for the biogenesis and development of RPs, providing new insights into rubber production mechanisms.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"16 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143666457","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}
Kai Feng, Jia-Lu Liu, Nan Sun, Zi-Qi Zhou, Zhi-Yuan Yang, Hui Lv, Cheng Yao, Jin-Ping Zou, Shu-Ping Zhao, Peng Wu, Liang-Jun Li
Releasing large quantities of volatiles is a defense strategy used by plants to resist herbivore attack. Oenanthe javanica, a perennial herb of the Apiaceae family, has a distinctive aroma due to volatile terpenoid accumulation. At present, the complete genome and genetic characteristics of volatile terpenoids in O. javanica remain largely unclear. Here, the telomere-to-telomere genome of O. javanica, with a size of 1012.13 Mb and a contig N50 of 49.55 Mb, was established by combining multiple sequencing technologies. Comparative genome analysis revealed that O. javanica experienced a recent species-specific whole-genome duplication event during the evolutionary process. Numerous gene family expansions were significantly enriched in the terpenoid biosynthesis process, monoterpenoid, and diterpenoid biosynthesis pathways, which resulted in abundant volatile substance accumulation in O. javanica. The volatile terpenoids of O. javanica showed repellent effects on herbivores. Terpenoid biosynthesis was activated by wounding signals under exogenous stimuli. The TPS gene family was significantly expanded in O. javanica compared to those in other species, and the members (OjTPS1, OjTPS3, OjTPS4, OjTPS5, OjTPS7, OjTPS16, OjTPS18, OjTPS30 and OjTPS58) responsible for different terpenoid biosynthesis were functionally characterized. These results reveal the genome evolution and molecular characteristics of volatile terpenoids in the process of plant–herbivore interactions. This study also provides genomic resources for genetic and molecular biology research on O. javanica and other plants.
{"title":"Telomere-to-telomere genome assembly reveals insights into the adaptive evolution of herbivore-defense mediated by volatile terpenoids in Oenanthe javanica","authors":"Kai Feng, Jia-Lu Liu, Nan Sun, Zi-Qi Zhou, Zhi-Yuan Yang, Hui Lv, Cheng Yao, Jin-Ping Zou, Shu-Ping Zhao, Peng Wu, Liang-Jun Li","doi":"10.1111/pbi.70062","DOIUrl":"https://doi.org/10.1111/pbi.70062","url":null,"abstract":"Releasing large quantities of volatiles is a defense strategy used by plants to resist herbivore attack. <i>Oenanthe javanica</i>, a perennial herb of the Apiaceae family, has a distinctive aroma due to volatile terpenoid accumulation. At present, the complete genome and genetic characteristics of volatile terpenoids in <i>O. javanica</i> remain largely unclear. Here, the telomere-to-telomere genome of <i>O. javanica</i>, with a size of 1012.13 Mb and a contig N50 of 49.55 Mb, was established by combining multiple sequencing technologies. Comparative genome analysis revealed that <i>O. javanica</i> experienced a recent species-specific whole-genome duplication event during the evolutionary process. Numerous gene family expansions were significantly enriched in the terpenoid biosynthesis process, monoterpenoid, and diterpenoid biosynthesis pathways, which resulted in abundant volatile substance accumulation in <i>O. javanica.</i> The volatile terpenoids of <i>O. javanica</i> showed repellent effects on herbivores. Terpenoid biosynthesis was activated by wounding signals under exogenous stimuli. The TPS gene family was significantly expanded in <i>O. javanica</i> compared to those in other species, and the members (<i>OjTPS1</i>, <i>OjTPS3</i>, <i>OjTPS4</i>, <i>OjTPS5</i>, <i>OjTPS7</i>, <i>OjTPS16</i>, <i>OjTPS18</i>, <i>OjTPS30</i> and <i>OjTPS58</i>) responsible for different terpenoid biosynthesis were functionally characterized. These results reveal the genome evolution and molecular characteristics of volatile terpenoids in the process of plant–herbivore interactions. This study also provides genomic resources for genetic and molecular biology research on <i>O. javanica</i> and other plants.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"3 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143666456","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}
Chen Feng, Muhammad Azhar Hussain, Yan Zhao, Yuning Wang, Yuyan Song, Yaxin Li, Hongtao Gao, Yan Jing, Keheng Xu, Wenping Zhang, Yonggang Zhou, Haiyan Li
Soybean is one of the most important crops in the world. However, salt stress poses a major challenge to soybean growth and productivity. Therefore, unravelling the complex mechanisms governing salt tolerance in soybean is imperative for molecular breeding of salt-tolerant varieties to improve yield. Maintaining intracellular Na+/K+ homeostasis is one of the key factors for plant salt tolerance. Although some salt tolerance mechanisms involving Na+ exclusion have been well identified in plants, few studies have been conducted on how K+ influx controls soybean salt tolerance. Here, we characterized the function of soybean K+ channel gene GmAKT1 and identified GmCBL9-GmCIPK6 complex, which modulated GmAKT1-mediated K+ uptake under salt stress. Functional studies found that soybean lines GmAKT1 overexpressing increased K+ content and promoted salt tolerance, while CRISPR/Cas9-mediated disruption of GmAKT1 soybean lines decreased the K+ content and showed salt sensitivity. Furthermore, we identified that GmCIPK6 interacted with GmAKT1 and GmCBL9 interacted with GmCIPK6. In addition, Mn2+-Phos-tag assays proved that GmCIPK6 could phosphorylate GmAKT1. This collaborative activation of the GmCBL9-GmCIPK6-GmAKT1 module promoted K+ influx and enhanced soybean salt tolerance. Our findings reveal a new molecular mechanism in soybeans under salt stress and provide insights for cultivating new salt-tolerant soybean varieties by molecular breeding.
{"title":"GmAKT1-mediated K+ absorption positively modulates soybean salt tolerance by GmCBL9-GmCIPK6 complex","authors":"Chen Feng, Muhammad Azhar Hussain, Yan Zhao, Yuning Wang, Yuyan Song, Yaxin Li, Hongtao Gao, Yan Jing, Keheng Xu, Wenping Zhang, Yonggang Zhou, Haiyan Li","doi":"10.1111/pbi.70042","DOIUrl":"https://doi.org/10.1111/pbi.70042","url":null,"abstract":"Soybean is one of the most important crops in the world. However, salt stress poses a major challenge to soybean growth and productivity. Therefore, unravelling the complex mechanisms governing salt tolerance in soybean is imperative for molecular breeding of salt-tolerant varieties to improve yield. Maintaining intracellular Na<sup>+</sup>/K<sup>+</sup> homeostasis is one of the key factors for plant salt tolerance. Although some salt tolerance mechanisms involving Na<sup>+</sup> exclusion have been well identified in plants, few studies have been conducted on how K<sup>+</sup> influx controls soybean salt tolerance. Here, we characterized the function of soybean K<sup>+</sup> channel gene <i>GmAKT1</i> and identified GmCBL9-GmCIPK6 complex, which modulated GmAKT1-mediated K<sup>+</sup> uptake under salt stress. Functional studies found that soybean lines <i>GmAKT1</i> overexpressing increased K<sup>+</sup> content and promoted salt tolerance, while CRISPR/Cas9-mediated disruption of <i>GmAKT1</i> soybean lines decreased the K<sup>+</sup> content and showed salt sensitivity. Furthermore, we identified that GmCIPK6 interacted with GmAKT1 and GmCBL9 interacted with GmCIPK6. In addition, Mn<sup>2+</sup>-Phos-tag assays proved that GmCIPK6 could phosphorylate GmAKT1. This collaborative activation of the GmCBL9-GmCIPK6-GmAKT1 module promoted K<sup>+</sup> influx and enhanced soybean salt tolerance. Our findings reveal a new molecular mechanism in soybeans under salt stress and provide insights for cultivating new salt-tolerant soybean varieties by molecular breeding.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"15 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143666458","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}
Liyong Zhang, Venkatesh Bollina, Peng Gao, Isobel A. P. Parkin
Camelina sativa is an important polyploid oilseed crop with multiple favourable agronomic traits. Capturing the leaf transcriptome of 48 accessions of C. sativa suggests allelic variation for gene expression levels and notably sub-genome dominance, both of which could provide opportunities for crop improvement. Flowering time (FT) is a crucial factor affecting the overall yield of crops. However, our understanding of the molecular mechanisms underlying FT regulation in C. sativa are still limited, partly due to its complex allohexaploid genome. In this study, weighted gene co-expression network analysis (WGCNA), expression quantitative trait loci (eQTL) analysis and transcriptome-wide association study (TWAS) were employed to explore the FT diversity among 48 C. sativa accessions and dissect the underlying molecular basis. Our results revealed a FT-related co-expressed gene module highly enriched with SOC1 and SOC1-like genes and identified 10 significant FT-associated single nucleotide polymorphisms (SNPs) defining three haplotype groups; thus providing a molecular basis for future genetic improvements in C. sativa breeding.
{"title":"Dissecting the molecular basis of variability for flowering time in Camelina sativa","authors":"Liyong Zhang, Venkatesh Bollina, Peng Gao, Isobel A. P. Parkin","doi":"10.1111/pbi.70049","DOIUrl":"https://doi.org/10.1111/pbi.70049","url":null,"abstract":"<i>Camelina sativa</i> is an important polyploid oilseed crop with multiple favourable agronomic traits. Capturing the leaf transcriptome of 48 accessions of <i>C. sativa</i> suggests allelic variation for gene expression levels and notably sub-genome dominance, both of which could provide opportunities for crop improvement. Flowering time (FT) is a crucial factor affecting the overall yield of crops. However, our understanding of the molecular mechanisms underlying FT regulation in <i>C. sativa</i> are still limited, partly due to its complex allohexaploid genome. In this study, weighted gene co-expression network analysis (WGCNA), expression quantitative trait loci (eQTL) analysis and transcriptome-wide association study (TWAS) were employed to explore the FT diversity among 48 <i>C. sativa</i> accessions and dissect the underlying molecular basis. Our results revealed a FT-related co-expressed gene module highly enriched with <i>SOC1</i> and <i>SOC1</i>-like genes and identified 10 significant FT-associated single nucleotide polymorphisms (SNPs) defining three haplotype groups; thus providing a molecular basis for future genetic improvements in <i>C. sativa</i> breeding.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"14 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143660375","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}
Yijie Zhan, Yumei Xia, Yao Wang, Siqing Liu, XiuLi Zhang, Shuo Xiong, Qiming Lv, Mengliang Cao
Significant progress in apomictic hybrid rice development faces challenges like achieving high induction rates and seed-setting efficiencies, and distinguishing clonal from zygotic embryos. To address the challenge of selecting clonal seeds, we developed a dual-fluorescence labelling gene switch system using the recombinase Cre/LoxP + FRT. Initially, this system was tested in callus tissue under a constitutive promoter; then, we replaced the promoter with a pollen-specific one to develop the pollen-specific gene switch (PSGS) system. The effectiveness of PSGS in rice pollen was subsequently validated. After confirming its functionality, we co-transformed the PSGS vectors with apomixis vectors in hybrid rice Yongyou 2640 (YE) and Yongyou 4949 (YS) using Agrobacterium-mediated transformation. Finally, we identified 18 MiMe mutants carrying the PSGS; the progeny of 16 lines were all red fluorescence seeds (zygotic embryo). Surprisingly, line L47-4 and L151-1 yielded 418 (n = 418) and 218 (n = 1279) non-fluorescent seeds in the T1 generation, respectively. The ploidy detection of non-fluorescent seeds showed that 57 (n = 68) and 64 (n = 72) were diploid in Line L47-4 and L151-1, individually. This phenomenon was reproducible in the T2 generation; 97 (n = 121) and 164 (n = 187) non-fluorescent seeds were diploid from line L47-4 and L151-1, respectively. This study demonstrates the ability of PSGS to distinguish between clonal seeds and zygotic seeds, with a sorting accuracy rate ranging from 80.2% to 88.9%, which is essential for improving clonal seed purity and advancing apomixis in rice cultivation.
{"title":"Efficient clonal seeds sorting for apomictic hybrid rice using a pollen-specific gene switch system","authors":"Yijie Zhan, Yumei Xia, Yao Wang, Siqing Liu, XiuLi Zhang, Shuo Xiong, Qiming Lv, Mengliang Cao","doi":"10.1111/pbi.70031","DOIUrl":"https://doi.org/10.1111/pbi.70031","url":null,"abstract":"Significant progress in apomictic hybrid rice development faces challenges like achieving high induction rates and seed-setting efficiencies, and distinguishing clonal from zygotic embryos. To address the challenge of selecting clonal seeds, we developed a dual-fluorescence labelling gene switch system using the recombinase <i>Cre/LoxP + FRT</i>. Initially, this system was tested in callus tissue under a constitutive promoter; then, we replaced the promoter with a pollen-specific one to develop the pollen-specific gene switch (PSGS) system. The effectiveness of PSGS in rice pollen was subsequently validated. After confirming its functionality, we co-transformed the PSGS vectors with apomixis vectors in hybrid rice Yongyou 2640 (YE) and Yongyou 4949 (YS) using <i>Agrobacterium</i>-mediated transformation. Finally, we identified 18 <i>MiMe</i> mutants carrying the PSGS; the progeny of 16 lines were all red fluorescence seeds (zygotic embryo). Surprisingly, line L47-4 and L151-1 yielded 418 (<i>n</i> = 418) and 218 (<i>n</i> = 1279) non-fluorescent seeds in the T<sub>1</sub> generation, respectively. The ploidy detection of non-fluorescent seeds showed that 57 (<i>n</i> = 68) and 64 (<i>n</i> = 72) were diploid in Line L47-4 and L151-1, individually. This phenomenon was reproducible in the T<sub>2</sub> generation; 97 (<i>n</i> = 121) and 164 (<i>n</i> = 187) non-fluorescent seeds were diploid from line L47-4 and L151-1, respectively. This study demonstrates the ability of PSGS to distinguish between clonal seeds and zygotic seeds, with a sorting accuracy rate ranging from 80.2% to 88.9%, which is essential for improving clonal seed purity and advancing apomixis in rice cultivation.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"200 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143660377","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}
Haomiao Yu, Jinqiu Liao, Yuanyuan Jiang, Mingzhi Zhong, Shan Tao, Songyue Chai, Long Wang, Li Lin, Ruiwu Yang, Xuexue Deng, Yunsong Zhang, Xiang Pu, Moyang Liu, Li Zhang
Salvia miltiorrhiza Bunge, a renowned medicinal herb in traditional Chinese medicine, displays distinctive root texture and high phenolic acid content, traits influenced by genetic and environmental factors. However, the underlying regulatory networks remain unclear. Here, we performed multi-omics analyses on ecotypes from four major Chinese regions, focusing on environmental impacts on root structure, phenolic acid accumulation and lignin composition. Lower temperatures and increased UV-B radiation were associated with elevated rosmarinic acid (RA) and salvianolic acid B (SAB) levels, particularly in the Sichuan ecotype. Structural models indicated that the radial arrangement of xylem conduits contributes to greater root hardness. Genomic assembly and comparative analysis of the Sichuan ecotype revealed a unique phenolic acid metabolism gene cluster, including SmWRKY40, a WRKY transcription factor essential for RA and SAB biosynthesis. Overexpression of SmWRKY40 enhanced phenolic acid levels and lignin content, whereas its knockout reduced root hardness. Integrating high-throughput (DNA affinity purification sequencing) and point-to-point (Yeast One-Hybrid, Dual-Luciferase and Electrophoretic Mobility Shift Assay) protein-DNA interaction detection platform further identified SmWRKY40 binding sites across ecotypes, revealing specific regulatory networks. Our findings provide insights into the molecular basis of root texture and bioactive compound accumulation, advancing breeding strategies for quality improvement in S. miltiorrhiza.
{"title":"Ecotype-specific phenolic acid accumulation and root softness in Salvia miltiorrhiza are driven by environmental and genetic factors","authors":"Haomiao Yu, Jinqiu Liao, Yuanyuan Jiang, Mingzhi Zhong, Shan Tao, Songyue Chai, Long Wang, Li Lin, Ruiwu Yang, Xuexue Deng, Yunsong Zhang, Xiang Pu, Moyang Liu, Li Zhang","doi":"10.1111/pbi.70048","DOIUrl":"https://doi.org/10.1111/pbi.70048","url":null,"abstract":"<i>Salvia miltiorrhiza</i> Bunge, a renowned medicinal herb in traditional Chinese medicine, displays distinctive root texture and high phenolic acid content, traits influenced by genetic and environmental factors. However, the underlying regulatory networks remain unclear. Here, we performed multi-omics analyses on ecotypes from four major Chinese regions, focusing on environmental impacts on root structure, phenolic acid accumulation and lignin composition. Lower temperatures and increased UV-B radiation were associated with elevated rosmarinic acid (RA) and salvianolic acid B (SAB) levels, particularly in the Sichuan ecotype. Structural models indicated that the radial arrangement of xylem conduits contributes to greater root hardness. Genomic assembly and comparative analysis of the Sichuan ecotype revealed a unique phenolic acid metabolism gene cluster, including <i>SmWRKY40</i>, a WRKY transcription factor essential for RA and SAB biosynthesis. Overexpression of <i>SmWRKY40</i> enhanced phenolic acid levels and lignin content, whereas its knockout reduced root hardness. Integrating high-throughput (DNA affinity purification sequencing) and point-to-point (Yeast One-Hybrid, Dual-Luciferase and Electrophoretic Mobility Shift Assay) protein-DNA interaction detection platform further identified <i>SmWRKY40</i> binding sites across ecotypes, revealing specific regulatory networks. Our findings provide insights into the molecular basis of root texture and bioactive compound accumulation, advancing breeding strategies for quality improvement in <i>S. miltiorrhiza</i>.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"43 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143660376","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}
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}
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