Li Chen, Keqin Chen, Jiapeng Jiang, Dan Wang, Kekun Zhang, Yulin Fang
SummaryWinter dormancy and bud break are crucial to the viability, adaptability and yield of fruit trees, but not all metabolic activities or regulatory factors involved in maintaining and breaking dormancy are known. Here, winter buds, spanning from natural dormancy to bud break, were collected from ‘Cabernet Sauvignon’ grapevines maintained outdoors or forced indoors. The transcriptomes, proteomes and plant hormone contents were analysed across several bud stages. The winter buds presented three main stages, dormancy, dormancy release and bud development, whether grown in or outdoors. Weighted Correlation Network Analysis (WGCNA) and Gene Ontology (GO) analysis of the omics data revealed that the different stages were enriched for different biological processes. Analysis of the differentially expressed genes (DEGs) identified seven candidate genes that may affect grape dormancy and bud break. Transient transformation of these seven genes showed that VvDOGL4, VvAGL65 and VvMARD could promote maintenance of winter bud dormancy in grapevine. Subcellular localization showed that these three proteins all located to the nucleus, and yeast two‐hybrid screening showed that they may interact with proteins related to plant hormone signal transduction, respiration, energy metabolism and transcription regulation to affect winter bud break in grapevine. Overall, these findings contribute to a better understanding of the regulatory dynamics of bud dormancy in a perennial fruit crop and lay a foundation for exploring key genes and regulatory mechanisms that can be manipulated to improve fruit quality and yields as the global climate shifts growing regions.
{"title":"Multi‐omics analysis of the regulatory network in winter buds of ‘Cabernet Sauvignon’ grapevine from dormancy to bud break","authors":"Li Chen, Keqin Chen, Jiapeng Jiang, Dan Wang, Kekun Zhang, Yulin Fang","doi":"10.1111/pbi.70014","DOIUrl":"https://doi.org/10.1111/pbi.70014","url":null,"abstract":"SummaryWinter dormancy and bud break are crucial to the viability, adaptability and yield of fruit trees, but not all metabolic activities or regulatory factors involved in maintaining and breaking dormancy are known. Here, winter buds, spanning from natural dormancy to bud break, were collected from ‘Cabernet Sauvignon’ grapevines maintained outdoors or forced indoors. The transcriptomes, proteomes and plant hormone contents were analysed across several bud stages. The winter buds presented three main stages, dormancy, dormancy release and bud development, whether grown in or outdoors. Weighted Correlation Network Analysis (<jats:styled-content style=\"fixed-case\">WGCNA</jats:styled-content>) and Gene Ontology (<jats:styled-content style=\"fixed-case\">GO</jats:styled-content>) analysis of the omics data revealed that the different stages were enriched for different biological processes. Analysis of the differentially expressed genes (<jats:styled-content style=\"fixed-case\">DEGs</jats:styled-content>) identified seven candidate genes that may affect grape dormancy and bud break. Transient transformation of these seven genes showed that <jats:styled-content style=\"fixed-case\"><jats:italic>VvDOGL4</jats:italic></jats:styled-content>, <jats:styled-content style=\"fixed-case\"><jats:italic>VvAGL65</jats:italic></jats:styled-content> and <jats:styled-content style=\"fixed-case\"><jats:italic>VvMARD</jats:italic></jats:styled-content> could promote maintenance of winter bud dormancy in grapevine. Subcellular localization showed that these three proteins all located to the nucleus, and yeast two‐hybrid screening showed that they may interact with proteins related to plant hormone signal transduction, respiration, energy metabolism and transcription regulation to affect winter bud break in grapevine. Overall, these findings contribute to a better understanding of the regulatory dynamics of bud dormancy in a perennial fruit crop and lay a foundation for exploring key genes and regulatory mechanisms that can be manipulated to improve fruit quality and yields as the global climate shifts growing regions.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"39 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143618479","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}
Xinyang Wang, Runfeng Wang, Xing Huo, Yueni Zhou, Muhammad J. Umer, Zihao Zheng, Weicai Jin, Lu Huang, Haifen Li, Qianxia Yu, Shaoxiong Li, Rajeev K Varshney, Wenyi Wang, Yuan Xiao, Yanbin Hong, Xiaoping Chen, Qing Lu, Hao Liu
SummaryPeanut stem is a vital organ to provide mechanical support and energy for aerial tissue development. However, the transcriptional regulatory mechanisms underlying stem development at a single‐cell resolution remain unclear. Herein, single‐nuclei isolation coupled with fluorescent‐activated cell sorting was employed to construct a cell atlas of peanut seedling stems using microdroplets‐based single‐nuclei RNA‐sequencing. This approach yielded 29 308 cells with 53 349 expressed genes underlying the identification of five cell types characterized by known marker genes. Additionally, 2053 differentially expressed genes (DEGs) were identified across different cell types. Furthermore, 3306 core‐DEGs involved in cell development trajectories were used to construct a transcription factor (TF) interaction network, providing insights into specific biological pathways and transcriptional regulation dynamics underlying cell‐type differentiation. Additionally, 1446 DEGs associated with different cell‐cycle profile were identified, revealing that peanut stem elongation and cell expansion are closely linked to auxin‐responsive pathway. This was supported by the examination of endogenous phytohormones and the identification of 10 hormone‐responsive DEGs. Moreover, AhWRKY70 was localized in the nucleus and is highly enriched in stem cortex and xylem cells and exhibits a tissue‐specific expression pattern that regulates stem growth. Overexpression of AhWRKY70 in Arabidopsis led to accelerated stem growth by modulating the phytohormone signalling pathway, influencing the expression of sixteen auxin and ethylene‐responsive genes as demonstrated by transcriptome sequencing. In conclusion, the single‐cell atlas provides a foundational dataset for understanding gene expression heterogeneity in peanut seedling stems. The elucidation of AhWRKY70 function expands our understanding of the roles of WRKY family members in peanut.
{"title":"Integration of single‐nuclei transcriptome and bulk RNA‐seq to unravel the role of AhWRKY70 in regulating stem cell development in Arachis hypogaea L.","authors":"Xinyang Wang, Runfeng Wang, Xing Huo, Yueni Zhou, Muhammad J. Umer, Zihao Zheng, Weicai Jin, Lu Huang, Haifen Li, Qianxia Yu, Shaoxiong Li, Rajeev K Varshney, Wenyi Wang, Yuan Xiao, Yanbin Hong, Xiaoping Chen, Qing Lu, Hao Liu","doi":"10.1111/pbi.70009","DOIUrl":"https://doi.org/10.1111/pbi.70009","url":null,"abstract":"SummaryPeanut stem is a vital organ to provide mechanical support and energy for aerial tissue development. However, the transcriptional regulatory mechanisms underlying stem development at a single‐cell resolution remain unclear. Herein, single‐nuclei isolation coupled with fluorescent‐activated cell sorting was employed to construct a cell atlas of peanut seedling stems using microdroplets‐based single‐nuclei RNA‐sequencing. This approach yielded 29 308 cells with 53 349 expressed genes underlying the identification of five cell types characterized by known marker genes. Additionally, 2053 differentially expressed genes (DEGs) were identified across different cell types. Furthermore, 3306 core‐DEGs involved in cell development trajectories were used to construct a transcription factor (TF) interaction network, providing insights into specific biological pathways and transcriptional regulation dynamics underlying cell‐type differentiation. Additionally, 1446 DEGs associated with different cell‐cycle profile were identified, revealing that peanut stem elongation and cell expansion are closely linked to auxin‐responsive pathway. This was supported by the examination of endogenous phytohormones and the identification of 10 hormone‐responsive DEGs. Moreover, <jats:italic>AhWRKY70</jats:italic> was localized in the nucleus and is highly enriched in stem cortex and xylem cells and exhibits a tissue‐specific expression pattern that regulates stem growth. Overexpression of <jats:italic>AhWRKY70</jats:italic> in <jats:italic>Arabidopsis</jats:italic> led to accelerated stem growth by modulating the phytohormone signalling pathway, influencing the expression of sixteen auxin and ethylene‐responsive genes as demonstrated by transcriptome sequencing. In conclusion, the single‐cell atlas provides a foundational dataset for understanding gene expression heterogeneity in peanut seedling stems. The elucidation of <jats:italic>AhWRKY70</jats:italic> function expands our understanding of the roles of <jats:italic>WRKY</jats:italic> family members in peanut.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"8 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143618423","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}
SummarySoil salinity is a severe abiotic stress that damages plant growth and development. As an antioxidant and free radical scavenger, melatonin is well known for helping plants survive abiotic conditions, including salinity stress. Here, we report that the salt‐related gene MsSNAT1, encoding a rate‐limiting melatonin biosynthesis enzyme, is located in the chloroplast and contributes to salinity stress tolerance in alfalfa. We found that the MsSNAT1 overexpressing alfalfa lines exhibited higher endogenous melatonin levels and increased tolerance to salt stress by promoting antioxidant systems and improving ion homeostasis. Furthermore, through a combination of transcriptome sequencing, dual‐luciferase assays and transgenic analysis, we identified that the basic leucine zipper (bZIP) transcription factor, MsbZIP55, is associated with salt response and MsSNAT1 expression. EMSA analysis and ChIP‐qPCR uncovered that MsbZIP55 can recognize and directly bind to the MsSNAT1 promoter in vitro and in vivo. MsbZIP55 acts as a negative regulator of MsSNAT1 expression, thereby reducing melatonin biosynthesis. Morphological analysis revealed that overexpressing MsbZIP55 conferred salt sensitivity to transgenic alfalfa through a higher Na+/K+ ratio and lower antioxidant activities, which could be alleviated by applying exogenous melatonin. Silencing of MsbZIP55 by RNA interference in alfalfa resulted in higher expression of MsSNAT1 and promoted salt tolerance by enhancing the antioxidant system enzyme activities and ion homeostasis. Our findings indicate that the MsbZIP55‐MsSNAT1 module plays a crucial role in regulating melatonin biosynthesis in alfalfa while facilitating protection against salinity stress. These results shed light on the regulatory mechanism of melatonin biosynthesis related to the salinity stress response in alfalfa.
{"title":"MsbZIP55 regulates salinity tolerance by modulating melatonin biosynthesis in alfalfa","authors":"Tingting Wang, JiaQi Yang, JiaMin Cao, Qi Zhang, HuaYue Liu, Peng Li, YiZhi Huang, WenWu Qian, Xiaojing Bi, Hui Wang, Yunwei Zhang","doi":"10.1111/pbi.70035","DOIUrl":"https://doi.org/10.1111/pbi.70035","url":null,"abstract":"SummarySoil salinity is a severe abiotic stress that damages plant growth and development. As an antioxidant and free radical scavenger, melatonin is well known for helping plants survive abiotic conditions, including salinity stress. Here, we report that the salt‐related gene <jats:italic>MsSNAT1</jats:italic>, encoding a rate‐limiting melatonin biosynthesis enzyme, is located in the chloroplast and contributes to salinity stress tolerance in alfalfa. We found that the <jats:italic>MsSNAT1</jats:italic> overexpressing alfalfa lines exhibited higher endogenous melatonin levels and increased tolerance to salt stress by promoting antioxidant systems and improving ion homeostasis. Furthermore, through a combination of transcriptome sequencing, dual‐luciferase assays and transgenic analysis, we identified that the basic leucine zipper (bZIP) transcription factor, MsbZIP55, is associated with salt response and <jats:italic>MsSNAT1</jats:italic> expression. EMSA analysis and ChIP‐qPCR uncovered that MsbZIP55 can recognize and directly bind to the <jats:italic>MsSNAT1</jats:italic> promoter <jats:italic>in vitro</jats:italic> and <jats:italic>in vivo</jats:italic>. MsbZIP55 acts as a negative regulator of <jats:italic>MsSNAT1</jats:italic> expression, thereby reducing melatonin biosynthesis. Morphological analysis revealed that overexpressing <jats:italic>MsbZIP55</jats:italic> conferred salt sensitivity to transgenic alfalfa through a higher Na<jats:sup>+</jats:sup>/K<jats:sup>+</jats:sup> ratio and lower antioxidant activities, which could be alleviated by applying exogenous melatonin. Silencing of <jats:italic>MsbZIP55</jats:italic> by RNA interference in alfalfa resulted in higher expression of <jats:italic>MsSNAT1</jats:italic> and promoted salt tolerance by enhancing the antioxidant system enzyme activities and ion homeostasis. Our findings indicate that the MsbZIP55‐MsSNAT1 module plays a crucial role in regulating melatonin biosynthesis in alfalfa while facilitating protection against salinity stress. These results shed light on the regulatory mechanism of melatonin biosynthesis related to the salinity stress response in alfalfa.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"18 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143618424","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}
SummaryThe dissection of genetic architecture for rice root system is largely dependent on phenotyping techniques, and high‐throughput root phenotyping poses a great challenge. In this study, we established a cost‐effective root phenotyping platform capable of analysing 1680 root samples within 2 h. To efficiently process a large number of root images, we developed the root phenotyping toolbox (RPT) with an enhanced SegFormer algorithm and used it for root segmentation and root phenotypic traits. Based on this root phenotyping platform and RPT, we screened 18 candidate (quantitative trait loci) QTL regions from 219 rice recombinant inbred lines under drought stress and validated the drought‐resistant functions of gene OsIAA8 identified from these QTL regions. This study confirmed that RPT exhibited a great application potential for processing images with various sources and for mining stress‐resistance genes of rice cultivars. Our developed root phenotyping platform and RPT software significantly improved high‐throughput root phenotyping efficiency, allowing for large‐scale root trait analysis, which will promote the genetic architecture improvement of drought‐resistant cultivars and crop breeding research in the future.
{"title":"RPT: An integrated root phenotyping toolbox for segmenting and quantifying root system architecture","authors":"Jiawei Shi, Shangyuan Xie, Weikun Li, Xin Wang, Jianglin Wang, Yunyu Chen, Yongyue Chang, Qiaojun Lou, Wanneng Yang","doi":"10.1111/pbi.70040","DOIUrl":"https://doi.org/10.1111/pbi.70040","url":null,"abstract":"SummaryThe dissection of genetic architecture for rice root system is largely dependent on phenotyping techniques, and high‐throughput root phenotyping poses a great challenge. In this study, we established a cost‐effective root phenotyping platform capable of analysing 1680 root samples within 2 h. To efficiently process a large number of root images, we developed the root phenotyping toolbox (RPT) with an enhanced SegFormer algorithm and used it for root segmentation and root phenotypic traits. Based on this root phenotyping platform and RPT, we screened 18 candidate (quantitative trait loci) QTL regions from 219 rice recombinant inbred lines under drought stress and validated the drought‐resistant functions of gene <jats:italic>OsIAA8</jats:italic> identified from these QTL regions. This study confirmed that RPT exhibited a great application potential for processing images with various sources and for mining stress‐resistance genes of rice cultivars. Our developed root phenotyping platform and RPT software significantly improved high‐throughput root phenotyping efficiency, allowing for large‐scale root trait analysis, which will promote the genetic architecture improvement of drought‐resistant cultivars and crop breeding research in the future.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"16 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143607961","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}
Isobel L. Dodds, Emma C. Watts, Mariana Schuster, Pierre Buscaill, Yasin Tumas, Nicholas J. Holton, Shijian Song, Johannes Stuttmann, Matthieu H. A. J. Joosten, Tolga Bozkurt, Renier A. L. van der Hoorn
<p>The infiltration of <i>Nicotiana benthamiana</i> with <i>Agrobacterium tumefaciens</i> (agroinfiltration) has become a routine expression platform for plant science and molecular pharming, yet this platform remains to be further optimized. We recently showed that <i>N. benthamiana</i> silenced for the <i>cold shock protein</i> (<i>CSP</i>) <i>receptor</i> (<i>CORE</i>) enables 8-fold more GFP production in older, 6–8-week-old plants, which are normally not used because of low transient expression efficiencies (Dodds <i>et al</i>., <span>2023</span>). Here, we investigated whether we can also increase transient protein expression levels in routinely used younger, 5-week-old juvenile plants, by silencing immunity-related genes.</p> <p>We selected 21 immunity-related genes encoding proteins that act at different levels in the plant immune system (Table S1). Besides <i>CORE</i>, we silenced receptor-encoding genes <i>WAK1</i> (<i>Wall-associated Protein Kinase</i>), <i>CERK1</i> (<i>Chitin Elicitor Receptor Kinase-1</i>), <i>BAK1</i> (<i>BRI1-associated Receptor Kinase-1</i>), <i>SOBIR1</i> (<i>Suppressor of BIR1-1</i>) and <i>RE02</i> (<i>Receptor of SCPs</i>). We also tested silencing of immune signaling components such as <i>F-box protein ACIF1</i> (<i>Avr/Cf-induced F-box-1</i>); lipase-like proteins <i>EDS1</i> (<i>Enhanced Disease Susceptibility-1</i>) and <i>SAG101</i> (<i>Senescence-associated Gene-101</i>); <i>CDPK</i> (<i>Calcium-dependent Protein Kinase</i>), <i>MPK3/6</i> (<i>MAP protein kinases-3 and -6</i>), <i>Nod-like helper receptors NRC2/3/4</i> (<i>NLR Required for Cf Signaling</i>) and chaperone <i>CRT3a</i> (<i>Calreticulin-3a</i>). We also included genes required for stress hormone signaling, including <i>PAL</i> (<i>Phenylalanine Ammonia Lyase</i>), <i>ICS</i> (<i>Isochorismate Synthase</i>), <i>NPR1</i> (<i>Nonexpressor of PR genes-1</i>), <i>EIN2</i> (<i>Ethylene-insensitive-2</i>) and <i>WRKY</i> transcription factors. Finally, we included genes encoding <i>AHA2</i> (<i>Arabidopsis H</i><sup>+</sup>-<i>ATPase 2</i>) and <i>RBOHB</i> (<i>Respiratory Burst Oxidase Homolog B</i>). Genes encoding phytoene desaturase (<i>PDS</i>) and <i>ß-glucuronidase</i> (<i>GUS</i>) were included as positive and negative controls for silencing, respectively. We resynthesized the silencing fragments as published previously (Table S1) and selected novel fragments targeting <i>ACIF1, CDPK, CORE, ICS</i> and <i>AHA2</i> (Dodds <i>et al</i>., <span>2023</span>, Tables S2 and S3).</p> <p>Tobacco Rattle Virus (TRV) vectors, each carrying a fragment of these 21 immunity genes and the controls were agroinfiltrated into 2-week-old seedlings and plants were tested for transient expression three weeks later. Transcript levels of the targeted genes were downregulated with novel silencing fragments (Figure S1). At that stage, no strong phenotypes were observed in TRV-inoculated plants, except for photobleaching in <i>TRV::PDS</i> plants, dwarfed <i>TRV::
{"title":"Immunity gene silencing increases transient protein expression in Nicotiana benthamiana","authors":"Isobel L. Dodds, Emma C. Watts, Mariana Schuster, Pierre Buscaill, Yasin Tumas, Nicholas J. Holton, Shijian Song, Johannes Stuttmann, Matthieu H. A. J. Joosten, Tolga Bozkurt, Renier A. L. van der Hoorn","doi":"10.1111/pbi.70005","DOIUrl":"https://doi.org/10.1111/pbi.70005","url":null,"abstract":"<p>The infiltration of <i>Nicotiana benthamiana</i> with <i>Agrobacterium tumefaciens</i> (agroinfiltration) has become a routine expression platform for plant science and molecular pharming, yet this platform remains to be further optimized. We recently showed that <i>N. benthamiana</i> silenced for the <i>cold shock protein</i> (<i>CSP</i>) <i>receptor</i> (<i>CORE</i>) enables 8-fold more GFP production in older, 6–8-week-old plants, which are normally not used because of low transient expression efficiencies (Dodds <i>et al</i>., <span>2023</span>). Here, we investigated whether we can also increase transient protein expression levels in routinely used younger, 5-week-old juvenile plants, by silencing immunity-related genes.</p>\u0000<p>We selected 21 immunity-related genes encoding proteins that act at different levels in the plant immune system (Table S1). Besides <i>CORE</i>, we silenced receptor-encoding genes <i>WAK1</i> (<i>Wall-associated Protein Kinase</i>), <i>CERK1</i> (<i>Chitin Elicitor Receptor Kinase-1</i>), <i>BAK1</i> (<i>BRI1-associated Receptor Kinase-1</i>), <i>SOBIR1</i> (<i>Suppressor of BIR1-1</i>) and <i>RE02</i> (<i>Receptor of SCPs</i>). We also tested silencing of immune signaling components such as <i>F-box protein ACIF1</i> (<i>Avr/Cf-induced F-box-1</i>); lipase-like proteins <i>EDS1</i> (<i>Enhanced Disease Susceptibility-1</i>) and <i>SAG101</i> (<i>Senescence-associated Gene-101</i>); <i>CDPK</i> (<i>Calcium-dependent Protein Kinase</i>), <i>MPK3/6</i> (<i>MAP protein kinases-3 and -6</i>), <i>Nod-like helper receptors NRC2/3/4</i> (<i>NLR Required for Cf Signaling</i>) and chaperone <i>CRT3a</i> (<i>Calreticulin-3a</i>). We also included genes required for stress hormone signaling, including <i>PAL</i> (<i>Phenylalanine Ammonia Lyase</i>), <i>ICS</i> (<i>Isochorismate Synthase</i>), <i>NPR1</i> (<i>Nonexpressor of PR genes-1</i>), <i>EIN2</i> (<i>Ethylene-insensitive-2</i>) and <i>WRKY</i> transcription factors. Finally, we included genes encoding <i>AHA2</i> (<i>Arabidopsis H</i><sup>+</sup>-<i>ATPase 2</i>) and <i>RBOHB</i> (<i>Respiratory Burst Oxidase Homolog B</i>). Genes encoding phytoene desaturase (<i>PDS</i>) and <i>ß-glucuronidase</i> (<i>GUS</i>) were included as positive and negative controls for silencing, respectively. We resynthesized the silencing fragments as published previously (Table S1) and selected novel fragments targeting <i>ACIF1, CDPK, CORE, ICS</i> and <i>AHA2</i> (Dodds <i>et al</i>., <span>2023</span>, Tables S2 and S3).</p>\u0000<p>Tobacco Rattle Virus (TRV) vectors, each carrying a fragment of these 21 immunity genes and the controls were agroinfiltrated into 2-week-old seedlings and plants were tested for transient expression three weeks later. Transcript levels of the targeted genes were downregulated with novel silencing fragments (Figure S1). At that stage, no strong phenotypes were observed in TRV-inoculated plants, except for photobleaching in <i>TRV::PDS</i> plants, dwarfed <i>TRV::","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"89 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143618997","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}
Dan Su, Mengbo Wu, Hsihua Wang, Peng Shu, Haiyan Song, Heng Deng, Shizhe Yu, Pedro Garcia‐Caparros, Mondher Bouzayen, Yang Zhang, Mingchun Liu
SummaryFlavonoids are polyphenolic secondary metabolites in tomato fruit with important roles in nutritional quality. Dissecting the transcriptional regulatory network modulating flavonoid metabolism is the first step to improve the nutritional quality of tomato fruits through molecular breeding technology. In this study, we identified a transcription factor SlbHLH95 as a key regulator in flavonoid metabolism through analysis of the MicroTom Metabolic Network (MMN) data set. Functional analyses revealed that knockout of SlbHLH95 increased the accumulation of naringenin, while the levels of rutin and nictoflorin decreased. Conversely, overexpression of SlbHLH95 resulted in an opposite pattern of accumulation of flavonoids. Transactivation assays showed that SlbHLH95 positively activated the expression of SlF3H and SlFLS, two key enzyme‐encoding genes in the flavonoid pathway, while repressing the expression of SlCHS1. Electrophoretic mobility shift assays (EMSA) demonstrated that SlbHLH95 could directly bind to the promoters of SlF3H and SlFLS, although it could not bind to the promoter of SlCHS1. Furthermore, SlbHLH95 interacted with the transcription factor SlMYB12 and coordinately regulated the expression of SlF3H and SlFLS. Beyond its role in flavonoid metabolism, SlbHLH95 positively regulated the grey mould resistance in tomato fruits by repressing SlBG10. Overall, our findings revealed the important role of bi‐functional SlbHLH95 in flavonoid metabolism and grey mould resistance in tomato fruits by acting as both a transcriptional activator and a repressor. This study provides new insights into strategies for improving fruit quality and enhancing fruit disease resistance through targeted genetic modulation.
{"title":"Bi‐functional transcription factor SlbHLH95 regulates fruits flavonoid metabolism and grey mould resistance in tomato","authors":"Dan Su, Mengbo Wu, Hsihua Wang, Peng Shu, Haiyan Song, Heng Deng, Shizhe Yu, Pedro Garcia‐Caparros, Mondher Bouzayen, Yang Zhang, Mingchun Liu","doi":"10.1111/pbi.70033","DOIUrl":"https://doi.org/10.1111/pbi.70033","url":null,"abstract":"SummaryFlavonoids are polyphenolic secondary metabolites in tomato fruit with important roles in nutritional quality. Dissecting the transcriptional regulatory network modulating flavonoid metabolism is the first step to improve the nutritional quality of tomato fruits through molecular breeding technology. In this study, we identified a transcription factor SlbHLH95 as a key regulator in flavonoid metabolism through analysis of the MicroTom Metabolic Network (MMN) data set. Functional analyses revealed that knockout of <jats:italic>SlbHLH95</jats:italic> increased the accumulation of naringenin, while the levels of rutin and nictoflorin decreased. Conversely, overexpression of <jats:italic>SlbHLH95</jats:italic> resulted in an opposite pattern of accumulation of flavonoids. Transactivation assays showed that SlbHLH95 positively activated the expression of <jats:italic>SlF3H</jats:italic> and <jats:italic>SlFLS</jats:italic>, two key enzyme‐encoding genes in the flavonoid pathway, while repressing the expression of <jats:italic>SlCHS1</jats:italic>. Electrophoretic mobility shift assays (EMSA) demonstrated that SlbHLH95 could directly bind to the promoters of <jats:italic>SlF3H</jats:italic> and <jats:italic>SlFLS</jats:italic>, although it could not bind to the promoter of <jats:italic>SlCHS1</jats:italic>. Furthermore, SlbHLH95 interacted with the transcription factor SlMYB12 and coordinately regulated the expression of <jats:italic>SlF3H</jats:italic> and <jats:italic>SlFLS</jats:italic>. Beyond its role in flavonoid metabolism, SlbHLH95 positively regulated the grey mould resistance in tomato fruits by repressing <jats:italic>SlBG10</jats:italic>. Overall, our findings revealed the important role of bi‐functional SlbHLH95 in flavonoid metabolism and grey mould resistance in tomato fruits by acting as both a transcriptional activator and a repressor. This study provides new insights into strategies for improving fruit quality and enhancing fruit disease resistance through targeted genetic modulation.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"39 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143599835","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}
Chaohui Yan, Wandi Liu, Ruimin Li, Guotian Liu, Yuejin Wang
Grapes, as one of the world's oldest economic crops, are severely affected by grape powdery mildew, causing significant economic losses. As a phytoalexin against powdery mildew, stilbenes and their key synthetic gene, stilbene synthase (STS), are highly sought after by researchers. In our previous research, a new gene, VqNSTS2, was identified from Vitis quinquangularis accession 'Danfeng-2' through transcriptomic analysis. However, the function and molecular mechanism of VqNSTS2 gene remain unknown. Here, by characterization and transient overexpression of VqNSTS2, we demonstrated that its expression product, stilbenes, can be detected in the model plant tobacco, which does not inherently contain STSs. After artificially inoculating transgenic Arabidopsis lines overexpressing VqNSTS2 with Erysiphe necator, it was found that VqNSTS2 actively moved to the pathogen's haustorium after responding to the pathogen, recognized and enveloped the haustorium, blocking the pathogen's infection and invasion and exhibited disease resistance. Furthermore, Agrobacterium-mediated stable overexpression of VqNSTS2 promoted stilbene accumulation and enhanced resistance of the V. vinifera susceptible cultivar 'Thompson Seedless' to E. necator. Additionally, through screening and identification, a transcription factor, VqERF062, was found to directly bind to the DRE and RAA motifs on ProVqNSTS2, positively regulating VqNSTS2 expression. Moreover, VqERF062 directly interacted with VqERF1B to promote the transcription of VqNSTS2 in addition to forming a homodimer with itself. Taken together, our findings reveal that the VqERF1B-VqERF062- module is required for grape resistance to E. necator and providing insights into the regulatory mechanism of stilbenes biosynthesis.
{"title":"VqERF1B-VqERF062-VqNSTS2 transcriptional cascade enhances stilbene biosynthesis and resistance to powdery mildew in grapevine","authors":"Chaohui Yan, Wandi Liu, Ruimin Li, Guotian Liu, Yuejin Wang","doi":"10.1111/pbi.70041","DOIUrl":"https://doi.org/10.1111/pbi.70041","url":null,"abstract":"Grapes, as one of the world's oldest economic crops, are severely affected by grape powdery mildew, causing significant economic losses. As a phytoalexin against powdery mildew, stilbenes and their key synthetic gene, <i>stilbene synthase</i> (<i>STS</i>), are highly sought after by researchers. In our previous research, a new gene, <i>VqNSTS2</i>, was identified from <i>Vitis quinquangularis</i> accession 'Danfeng-2' through transcriptomic analysis. However, the function and molecular mechanism of <i>VqNSTS2</i> gene remain unknown. Here, by characterization and transient overexpression of <i>VqNSTS2</i>, we demonstrated that its expression product, stilbenes, can be detected in the model plant tobacco, which does not inherently contain <i>STSs</i>. After artificially inoculating transgenic Arabidopsis lines overexpressing <i>VqNSTS2</i> with <i>Erysiphe necator</i>, it was found that <i>VqNSTS2</i> actively moved to the pathogen's haustorium after responding to the pathogen, recognized and enveloped the haustorium, blocking the pathogen's infection and invasion and exhibited disease resistance. Furthermore, <i>Agrobacterium</i>-mediated stable overexpression of <i>VqNSTS2</i> promoted stilbene accumulation and enhanced resistance of the <i>V. vinifera</i> susceptible cultivar 'Thompson Seedless' to <i>E. necator</i>. Additionally, through screening and identification, a transcription factor, VqERF062, was found to directly bind to the DRE and RAA motifs on ProVqNSTS2, positively regulating <i>VqNSTS2</i> expression. Moreover, VqERF062 directly interacted with VqERF1B to promote the transcription of <i>VqNSTS2</i> in addition to forming a homodimer with itself. Taken together, our findings reveal that the VqERF1B-VqERF062- module is required for grape resistance to <i>E. necator</i> and providing insights into the regulatory mechanism of stilbenes biosynthesis.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"192 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143582641","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}
Friedrich Kauder, Gabor Gyetvai, Klaus Schmidt, Daniel Stirnweis, Tobias Haehre, Kai Prenzler, Anja Maeser, Christine Klapprodt, Florian Tiller, Jens Lübeck, Dietmar J. Stahl
Late blight resistance of potato was improved by the co-expression of the potato resistance gene R3a and the pathogen-inducible avirulence gene Avr3a of Phytopthora infestans. The synthetic pathogen-inducible promoter 2xS-4xD-NpCABEcore, which is composed of the cis-acting elements S and D and the core promoter of the NpCABE gene, was developed for potato. By analysis of 20 core promoters from Solanacea species synthetic promoters of the 2xS-2xD-type were generated which differ in their background activity, strength and promoter inducibility. These data showed that the core promoter plays an important role for the architecture of a synthetic promoter and influences the specificity and strength beside the cis-acting element. The 2xS-2xD-NpCABEcore promoter was further improved by increasing the number of the cis-acting elements resulting in the 2xS-4xD-NpCABEcore promoter. Modified Avr3a alleles, which triggered less cell death than the Avr3aKI allele, were expressed with the optimized synthetic promoter in transgenic potatoes with an R3a gene. The transgenic lines showed less late blight symptoms and up to 60% reduction of sporangia in detached leaf assays. The absence of a negative plant phenotype in the greenhouse demonstrated that the balanced co-expression of a modified Avr3a gene under the control of an optimized synthetic promoter is a promising strategy to increase late blight resistance of potatoes. This concept might be as well applied to other crops since the co-expression of the R3a and Avr3aKI gene induced cell death in leaves of corn, wheat and soybean in a transient assay.
{"title":"Expression of a modified Avr3a gene under the control of a synthetic pathogen-inducible promoter leads to Phytophthora infestans resistance in potato","authors":"Friedrich Kauder, Gabor Gyetvai, Klaus Schmidt, Daniel Stirnweis, Tobias Haehre, Kai Prenzler, Anja Maeser, Christine Klapprodt, Florian Tiller, Jens Lübeck, Dietmar J. Stahl","doi":"10.1111/pbi.14615","DOIUrl":"https://doi.org/10.1111/pbi.14615","url":null,"abstract":"Late blight resistance of potato was improved by the co-expression of the potato resistance gene <i>R3a</i> and the pathogen-inducible avirulence gene <i>Avr3a</i> of <i>Phytopthora infestans</i>. The synthetic pathogen-inducible promoter 2xS-4xD-NpCABE<sub>core</sub>, which is composed of the <i>cis</i>-acting elements S and D and the core promoter of the <i>NpCABE</i> gene, was developed for potato. By analysis of 20 core promoters from Solanacea species synthetic promoters of the 2xS-2xD-type were generated which differ in their background activity, strength and promoter inducibility. These data showed that the core promoter plays an important role for the architecture of a synthetic promoter and influences the specificity and strength beside the <i>cis</i>-acting element. The 2xS-2xD-NpCABE<sub>core</sub> promoter was further improved by increasing the number of the <i>cis</i>-acting elements resulting in the 2xS-4xD-NpCABE<sub>core</sub> promoter. Modified <i>Avr3a</i> alleles, which triggered less cell death than the <i>Avr3a</i><sup>KI</sup> allele, were expressed with the optimized synthetic promoter in transgenic potatoes with an <i>R3a</i> gene. The transgenic lines showed less late blight symptoms and up to 60% reduction of sporangia in detached leaf assays. The absence of a negative plant phenotype in the greenhouse demonstrated that the balanced co-expression of a modified <i>Avr3a</i> gene under the control of an optimized synthetic promoter is a promising strategy to increase late blight resistance of potatoes. This concept might be as well applied to other crops since the co-expression of the <i>R3a</i> and <i>Avr3a</i><sup><i>KI</i></sup> gene induced cell death in leaves of corn, wheat and soybean in a transient assay.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"19 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143582642","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}
<p>With climate change increasing the frequency of extreme weather events, waterlogging has become a significant threat to agricultural production, especially in maize-growing regions. Waterlogging induces hypoxic conditions in the root zone, limiting maize growth and yield (Liang <i>et al</i>., <span>2020</span>; Pedersen <i>et al</i>., <span>2017</span>). Plants have evolved adaptive mechanisms, such as adventitious root (AR) formation and enhanced antioxidant activity, to cope with waterlogging stress (Pedersen <i>et al</i>., <span>2021</span>; Yamauchi <i>et al</i>., <span>2018</span>). However, the regulatory mechanisms in maize remain poorly understood.</p> <p>Group VII ethylene response factor proteins (ERFVIIs) are key regulators of waterlogging tolerance in model plants (Hartman <i>et al</i>., <span>2021</span>). Our previous work showed that <i>ZmEREB180</i>, a maize ERFVII, promotes waterlogging tolerance by enhancing AR formation and modulating antioxidant levels (Yu <i>et al</i>., <span>2019</span>). In this study, we cloned the full-length coding sequence of <i>ZmEREB180</i> and inserted it into the pM999 vector. The recombinants and empty vector were transiently expressed in isolated B73 leaf protoplasts, followed by a transient and simplified cleavage under targets and tag-mentation (tsCUT&Tag) assay (Liang <i>et al</i>., <span>2024</span>). A total of 4720 confident peaks corresponding to 3335 genes were identified (Table S1). Notably, 70.15% of these peaks were located in promoter regions, with 68.67% found in promoters less than 1 kb upstream (Figure 1a). The highest enrichment was observed at the transcription start site (Figure 1b). Motif analysis revealed the GCC-box (GCCGCC) as the highest scoring motif (E-value = 5.7 × 10<sup>−10</sup>). Compared with RNA-Seq data (Yu <i>et al</i>., <span>2019</span>) identified 421 genes that were differentially expressed in the <i>ZmEREB180</i> overexpression lines, under waterlogged conditions, and were directly bound by ZmEREB180 (Figure 1c; Table S2). We focused on genes involved in root development and antioxidant pathways. Lateral organ boundaries domain (LBD) proteins play pivotal roles in organ development. Two LBD genes, <i>ZmLBD5</i> and <i>ZmLBD38</i> (Table S2), were up-regulated in an overexpression line and under waterlogging conditions, in which <i>ZmLBD5</i> has been shown to promote AR formation (Feng <i>et al</i>., <span>2022</span>). Four antioxidant genes, including two glutathione-S-transferases (GST, <i>ZmGST8</i> and <i>ZmGST31</i>) and two peroxidases (POD, <i>ZmPOD12</i> and <i>ZmPOD55</i>), exhibited similar expression profiles (Table S2). The tsCUT&Tag data revealed significant peaks in the promoter of these genes (Figure 1d). Additionally, GCC-box motifs were located in these regions, suggesting direct regulation by ZmEREB180 under waterlogging conditions.</p> <figure><picture> <source media="(min-width: 1650px)" srcset="/cms/asset/e7ef2f07-4842-485e-aa
{"title":"ZmEREB180 modulates waterlogging tolerance in maize by regulating root development and antioxidant gene expression","authors":"Huanhuan Qi, Jing Wang, Xin Wang, Kun Liang, Meicheng Ke, Xueqing Zheng, Wenbin Tang, Ziyun Chen, Yinggen Ke, Pingfang Yang, Fazhan Qiu, Feng Yu","doi":"10.1111/pbi.70030","DOIUrl":"https://doi.org/10.1111/pbi.70030","url":null,"abstract":"<p>With climate change increasing the frequency of extreme weather events, waterlogging has become a significant threat to agricultural production, especially in maize-growing regions. Waterlogging induces hypoxic conditions in the root zone, limiting maize growth and yield (Liang <i>et al</i>., <span>2020</span>; Pedersen <i>et al</i>., <span>2017</span>). Plants have evolved adaptive mechanisms, such as adventitious root (AR) formation and enhanced antioxidant activity, to cope with waterlogging stress (Pedersen <i>et al</i>., <span>2021</span>; Yamauchi <i>et al</i>., <span>2018</span>). However, the regulatory mechanisms in maize remain poorly understood.</p>\u0000<p>Group VII ethylene response factor proteins (ERFVIIs) are key regulators of waterlogging tolerance in model plants (Hartman <i>et al</i>., <span>2021</span>). Our previous work showed that <i>ZmEREB180</i>, a maize ERFVII, promotes waterlogging tolerance by enhancing AR formation and modulating antioxidant levels (Yu <i>et al</i>., <span>2019</span>). In this study, we cloned the full-length coding sequence of <i>ZmEREB180</i> and inserted it into the pM999 vector. The recombinants and empty vector were transiently expressed in isolated B73 leaf protoplasts, followed by a transient and simplified cleavage under targets and tag-mentation (tsCUT&Tag) assay (Liang <i>et al</i>., <span>2024</span>). A total of 4720 confident peaks corresponding to 3335 genes were identified (Table S1). Notably, 70.15% of these peaks were located in promoter regions, with 68.67% found in promoters less than 1 kb upstream (Figure 1a). The highest enrichment was observed at the transcription start site (Figure 1b). Motif analysis revealed the GCC-box (GCCGCC) as the highest scoring motif (E-value = 5.7 × 10<sup>−10</sup>). Compared with RNA-Seq data (Yu <i>et al</i>., <span>2019</span>) identified 421 genes that were differentially expressed in the <i>ZmEREB180</i> overexpression lines, under waterlogged conditions, and were directly bound by ZmEREB180 (Figure 1c; Table S2). We focused on genes involved in root development and antioxidant pathways. Lateral organ boundaries domain (LBD) proteins play pivotal roles in organ development. Two LBD genes, <i>ZmLBD5</i> and <i>ZmLBD38</i> (Table S2), were up-regulated in an overexpression line and under waterlogging conditions, in which <i>ZmLBD5</i> has been shown to promote AR formation (Feng <i>et al</i>., <span>2022</span>). Four antioxidant genes, including two glutathione-S-transferases (GST, <i>ZmGST8</i> and <i>ZmGST31</i>) and two peroxidases (POD, <i>ZmPOD12</i> and <i>ZmPOD55</i>), exhibited similar expression profiles (Table S2). The tsCUT&Tag data revealed significant peaks in the promoter of these genes (Figure 1d). Additionally, GCC-box motifs were located in these regions, suggesting direct regulation by ZmEREB180 under waterlogging conditions.</p>\u0000<figure><picture>\u0000<source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/e7ef2f07-4842-485e-aa","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"86 2 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143582647","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}
<p>In rice agronomy, hybridization is a crucial method to augment crop productivity. The cornerstone of hybrid crop breeding is the utilization of male-sterile lines. Compared to the traditional three-line breeding system, the two-line breeding strategy, which leverages thermo-sensitive genic male sterility (TGMS) and photoperiod-sensitive genic male sterility (PGMS), offers significant benefits by expanding the genetic reservoir available for breeding programs. Currently, two-line hybrid rice occupies 44% of the total hybrid rice cultivation area. However, the availability of TGMS and PGMS germplasm and genetic resources remains severely restricted. Notably, TGMS lines originating from mutations at the <i>tms5</i> locus account for at least 83.8% of the two-line hybrid rice varieties in China (Zhang <i>et al</i>., <span>2022</span>).</p> <p>The leucine-rich repeat receptor kinase EMS1 and its ligand, TPD1 peptide, form a critical receptor–ligand complex indispensable for the proper development of the anther tapetum. Mutations in <i>ems1</i> and <i>tpd1</i> both result in sterility characterized by an absence of pollen. Previous research has elucidated that EMS1 and the brassinosteroid receptor BRI1 utilize common downstream signalling pathways, allowing for the functional substitution of their kinase domains (Zheng <i>et al</i>., <span>2019</span>). Among the various BRI1 mutant alleles, <i>bri1-301</i> is particularly notable due to the G-989-I substitution, which almost completely eliminates kinase activity both <i>in vitro</i> and <i>in vivo</i>; yet surprisingly, it only produces a mild dwarf phenotype compared to more severe or null <i>bri1</i> alleles (Xu <i>et al</i>., <span>2008</span>). The phenotypic severity and protein accumulation of <i>bri1-301</i> are modulated by temperature (Figure 1a), with accelerated degradation occurring at elevated temperatures through an unidentified pathway. At 22°C, bri1-301 protein accumulates normally, whereas at 29°C, its accumulation is markedly compromised (Figure 1b; Figure S1) (Lv <i>et al</i>., <span>2018</span>; Zhang <i>et al</i>., <span>2018</span>). Transgenic expression of GFP-tagged bri1-301 retains its sensitivity to high temperatures (Figure 1c). Comparative evaluation of in vitro autophosphorylation activities reveals that bri1-301 loses most of its autophosphorylation capability, and EMS1 demonstrates significantly weaker autophosphorylation activity compared to BRI1 (Figure 1d). Consequently, we attempted to introduce the bri1-301 mutation site into a chimeric EMS1-BRI1 receptor to preserve biological activity while imparting temperature sensitivity (Figure 1e). By employing the <i>EMS1</i> promoter to drive the expression of the EMS1-BRI1* construct in the <i>ems1</i> mutant background, we achieved notable phenotypic restoration at 22°C, characterized by the generation of pollen and fertile siliques. Contrarily, this phenotypic amelioration was unattainable at 29°C (Figure 1f,g). Prot
{"title":"Creation of thermosensitive male sterility line in rice via a temperature-sensitive mutation in receptor kinase","authors":"Qunwei Bai, Fenghua Li, Jiajia Zhang, Aixia Huang, Chenyu Shi, Hongyan Ren, Bowen Zheng","doi":"10.1111/pbi.70027","DOIUrl":"https://doi.org/10.1111/pbi.70027","url":null,"abstract":"<p>In rice agronomy, hybridization is a crucial method to augment crop productivity. The cornerstone of hybrid crop breeding is the utilization of male-sterile lines. Compared to the traditional three-line breeding system, the two-line breeding strategy, which leverages thermo-sensitive genic male sterility (TGMS) and photoperiod-sensitive genic male sterility (PGMS), offers significant benefits by expanding the genetic reservoir available for breeding programs. Currently, two-line hybrid rice occupies 44% of the total hybrid rice cultivation area. However, the availability of TGMS and PGMS germplasm and genetic resources remains severely restricted. Notably, TGMS lines originating from mutations at the <i>tms5</i> locus account for at least 83.8% of the two-line hybrid rice varieties in China (Zhang <i>et al</i>., <span>2022</span>).</p>\u0000<p>The leucine-rich repeat receptor kinase EMS1 and its ligand, TPD1 peptide, form a critical receptor–ligand complex indispensable for the proper development of the anther tapetum. Mutations in <i>ems1</i> and <i>tpd1</i> both result in sterility characterized by an absence of pollen. Previous research has elucidated that EMS1 and the brassinosteroid receptor BRI1 utilize common downstream signalling pathways, allowing for the functional substitution of their kinase domains (Zheng <i>et al</i>., <span>2019</span>). Among the various BRI1 mutant alleles, <i>bri1-301</i> is particularly notable due to the G-989-I substitution, which almost completely eliminates kinase activity both <i>in vitro</i> and <i>in vivo</i>; yet surprisingly, it only produces a mild dwarf phenotype compared to more severe or null <i>bri1</i> alleles (Xu <i>et al</i>., <span>2008</span>). The phenotypic severity and protein accumulation of <i>bri1-301</i> are modulated by temperature (Figure 1a), with accelerated degradation occurring at elevated temperatures through an unidentified pathway. At 22°C, bri1-301 protein accumulates normally, whereas at 29°C, its accumulation is markedly compromised (Figure 1b; Figure S1) (Lv <i>et al</i>., <span>2018</span>; Zhang <i>et al</i>., <span>2018</span>). Transgenic expression of GFP-tagged bri1-301 retains its sensitivity to high temperatures (Figure 1c). Comparative evaluation of in vitro autophosphorylation activities reveals that bri1-301 loses most of its autophosphorylation capability, and EMS1 demonstrates significantly weaker autophosphorylation activity compared to BRI1 (Figure 1d). Consequently, we attempted to introduce the bri1-301 mutation site into a chimeric EMS1-BRI1 receptor to preserve biological activity while imparting temperature sensitivity (Figure 1e). By employing the <i>EMS1</i> promoter to drive the expression of the EMS1-BRI1* construct in the <i>ems1</i> mutant background, we achieved notable phenotypic restoration at 22°C, characterized by the generation of pollen and fertile siliques. Contrarily, this phenotypic amelioration was unattainable at 29°C (Figure 1f,g). Prot","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"25 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143582645","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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