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NbNAC1 enhances plant immunity against TMV by regulating isochorismate synthase 1 expression and the SA pathway
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.17242
Feng Zhu, Kainan Li, Mengyao Cao, Qiping Zhang, Yangkai Zhou, Huan Chen, Maha AlKhazindar, Zhaolin Ji

Salicylic acid (SA) plays important roles in plant local and systemic resistance. Isochorismate synthase 1 (ICS1) is a key enzyme in SA synthesis. Pathogens infection triggered the ICS1 expression and induced SA production. However, the molecular regulation mechanism of ICS1 against virus infection remains unclear. Here, we employed molecular genetics and physiobiochemical approaches to confirm a transcription factor NbNAC1 from Nicotiana benthamiana is a positive regulator of resistance against tobacco mosaic virus (TMV). The pathways NbNAC1 and NbICS1 can be triggered by TMV infection. Silencing NbNAC1 accelerated TMV-induced oxidative damage and increased reactive oxygen species (ROS) production. It also weakened both local and systemic resistance against TMV and decreased the expression of NbICS1, SA signaling gene NbNPR1, and SA defense-related genes. The effects of NbNAC1-silencing were restored by overexpression of NbICS1 or foliar SA applications. Overexpressing NbNAC1 prevented oxidative damage and reduced the production of ROS, enhanced plant resistance against viral pathogen, and activated NbICS1 expression, and SA downstream signaling and defense-related genes. NbNAC1 localized in nuclear and emerged the ability of transcriptional regulation. ChIP and EMSA results indicated that NbNAC1 directly binds to a fragment containing GAAATT motif of NbICS1 promoter. Luciferase reporter assays confirmed that NbNAC1 activates NbICS1 expression. Taken together, our results demonstrate that NbNAC1 plays a critical role in plant immunity through activation of SA production.

{"title":"NbNAC1 enhances plant immunity against TMV by regulating isochorismate synthase 1 expression and the SA pathway","authors":"Feng Zhu,&nbsp;Kainan Li,&nbsp;Mengyao Cao,&nbsp;Qiping Zhang,&nbsp;Yangkai Zhou,&nbsp;Huan Chen,&nbsp;Maha AlKhazindar,&nbsp;Zhaolin Ji","doi":"10.1111/tpj.17242","DOIUrl":"https://doi.org/10.1111/tpj.17242","url":null,"abstract":"<div>\u0000 \u0000 <p>Salicylic acid (SA) plays important roles in plant local and systemic resistance. Isochorismate synthase 1 (ICS1) is a key enzyme in SA synthesis. Pathogens infection triggered the <i>ICS1</i> expression and induced SA production. However, the molecular regulation mechanism of <i>ICS1</i> against virus infection remains unclear. Here, we employed molecular genetics and physiobiochemical approaches to confirm a transcription factor <i>NbNAC1</i> from <i>Nicotiana benthamiana</i> is a positive regulator of resistance against tobacco mosaic virus (TMV). The pathways <i>NbNAC1</i> and <i>NbICS1</i> can be triggered by TMV infection. Silencing <i>NbNAC1</i> accelerated TMV-induced oxidative damage and increased reactive oxygen species (ROS) production. It also weakened both local and systemic resistance against TMV and decreased the expression of <i>NbICS1</i>, SA signaling gene <i>NbNPR1</i>, and SA defense-related genes. The effects of <i>NbNAC1</i>-silencing were restored by overexpression of <i>NbICS1</i> or foliar SA applications. Overexpressing <i>NbNAC1</i> prevented oxidative damage and reduced the production of ROS, enhanced plant resistance against viral pathogen, and activated <i>NbICS1</i> expression, and SA downstream signaling and defense-related genes. NbNAC1 localized in nuclear and emerged the ability of transcriptional regulation. ChIP and EMSA results indicated that NbNAC1 directly binds to a fragment containing GAAATT motif of <i>NbICS1</i> promoter. Luciferase reporter assays confirmed that <i>NbNAC1</i> activates <i>NbICS1</i> expression. Taken together, our results demonstrate that <i>NbNAC1</i> plays a critical role in plant immunity through activation of SA production.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Unraveling the role of OsSCL26 in transcriptional regulation in rice: Insights into grain shape, heading date, and carbohydrates
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.17268
Xiaoli Jin, Yingying Lu, Jialin Liu, Hui Liu, Nan Wu, Mei Li, Weijun Zhou

Grain shape, heading date, and amylase content are pivotal traits influencing rice yield, quality, distribution, and regional adaptability. Through our investigation, we identified a mutant, characterized by slender grains, elevated amylose content, and early heading date. Histocytologic scrutiny unveiled heightened cell proliferation in the spikelet hull contributing to the slender grain morphology. The OsSCL26 gene, governing these significant traits, was meticulously cloned via fine-mapping. Phenotypic scrutiny of OsSCL26 knockout and overexpression lines validated its pivotal role in trait regulation. Further analysis disclosed a substitution in the OsSCL26 promoter region, creating a novel binding site for the transcript factor OsbZIP47, thereby modulating its expression in the osscl26 mutant. Functionally, OsSCL26, acting as a serine/arginine-rich SC35-like protein, interacted with U1-70K in vivo and in vitro. OsSCL26 exhibited direct binding to genes implicated in grain shape and carbohydrates, thereby regulating their splicing. Moreover, OsSCL26 showed direct and indirect associations with target RNAs involved in circadian rhythm. Overall, our findings elucidate the mechanism of OsSCL26, an RNA binding protein interacting with splicing factor, as a crucial member of the spliceosome, thereby impacting post-transcriptional splicing and regulating grain shape, heading date, and carbohydrates in rice.

{"title":"Unraveling the role of OsSCL26 in transcriptional regulation in rice: Insights into grain shape, heading date, and carbohydrates","authors":"Xiaoli Jin,&nbsp;Yingying Lu,&nbsp;Jialin Liu,&nbsp;Hui Liu,&nbsp;Nan Wu,&nbsp;Mei Li,&nbsp;Weijun Zhou","doi":"10.1111/tpj.17268","DOIUrl":"https://doi.org/10.1111/tpj.17268","url":null,"abstract":"<div>\u0000 \u0000 <p>Grain shape, heading date, and amylase content are pivotal traits influencing rice yield, quality, distribution, and regional adaptability. Through our investigation, we identified a mutant, characterized by slender grains, elevated amylose content, and early heading date. Histocytologic scrutiny unveiled heightened cell proliferation in the spikelet hull contributing to the slender grain morphology. The <i>OsSCL26</i> gene, governing these significant traits, was meticulously cloned via fine-mapping. Phenotypic scrutiny of <i>OsSCL26</i> knockout and overexpression lines validated its pivotal role in trait regulation. Further analysis disclosed a substitution in the <i>OsSCL26</i> promoter region, creating a novel binding site for the transcript factor OsbZIP47, thereby modulating its expression in the <i>osscl26</i> mutant. Functionally, OsSCL26, acting as a serine/arginine-rich SC35-like protein, interacted with U1-70K <i>in vivo</i> and in <i>vitro</i>. OsSCL26 exhibited direct binding to genes implicated in grain shape and carbohydrates, thereby regulating their splicing. Moreover, OsSCL26 showed direct and indirect associations with target RNAs involved in circadian rhythm. Overall, our findings elucidate the mechanism of OsSCL26, an RNA binding protein interacting with splicing factor, as a crucial member of the spliceosome, thereby impacting post-transcriptional splicing and regulating grain shape, heading date, and carbohydrates in rice.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Subterranean morphology underpins the degree of mycoheterotrophy, mycorrhizal associations, and plant vigor in a green orchid Oreorchis patens
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.70045
Kenji Suetsugu, Hidehito Okada

The evolution of full heterotrophy is a fascinating topic in plant evolution, with recent studies suggesting that partial mycoheterotrophy (mixotrophy) serves as a transitional stage toward full mycoheterotrophy in orchids. However, the adaptive significance of fungal-derived carbon in mixotrophic plants remains largely unexplored. In this study, we investigated the photosynthetic orchid Oreorchis patens, a species related to the leafless genus Corallorhiza within the subtribe Calypsoinae. Using high-throughput DNA sequencing, 13C and 15N isotopic analyses, and phenotypic evaluations, we explored the role of coralloid rhizomes – a feature common in fully mycoheterotrophic orchids – in fungal partnerships, the degree of mycoheterotrophy, and plant vigor. Our findings reveal that O. patens plants with coralloid rhizomes predominantly associate with saprotrophic Psathyrellaceae fungi, whereas those without coralloid rhizomes also partner with rhizoctonias and other potentially orchid mycorrhizal fungi. Notably, plants with coralloid rhizomes exhibited enriched 13C signatures, indicating a greater reliance on fungal-derived carbon. These plants also demonstrated more vigorous flowering scapes and produced a higher number of flowers, suggesting that mycoheterotrophy significantly enhances plant vigor. This study provides rare insights into the adaptive significance of mycoheterotrophy. Recent research suggests that some partially mycoheterotrophic orchids can adjust their heterotrophic status to optimize carbon resource use under specific conditions, such as low-light environments. However, an increased proportion of fungal-derived carbon may sometimes merely reflect reduced photosynthesis in such conditions, thereby amplifying the apparent contribution of fungal-derived carbon. Our findings offer more direct evidence that carbon acquisition via mycoheterotrophy is beneficial for partially mycoheterotrophic orchids.

{"title":"Subterranean morphology underpins the degree of mycoheterotrophy, mycorrhizal associations, and plant vigor in a green orchid Oreorchis patens","authors":"Kenji Suetsugu,&nbsp;Hidehito Okada","doi":"10.1111/tpj.70045","DOIUrl":"https://doi.org/10.1111/tpj.70045","url":null,"abstract":"<p>The evolution of full heterotrophy is a fascinating topic in plant evolution, with recent studies suggesting that partial mycoheterotrophy (mixotrophy) serves as a transitional stage toward full mycoheterotrophy in orchids. However, the adaptive significance of fungal-derived carbon in mixotrophic plants remains largely unexplored. In this study, we investigated the photosynthetic orchid <i>Oreorchis patens</i>, a species related to the leafless genus <i>Corallorhiza</i> within the subtribe Calypsoinae. Using high-throughput DNA sequencing, <sup>13</sup>C and <sup>15</sup>N isotopic analyses, and phenotypic evaluations, we explored the role of coralloid rhizomes – a feature common in fully mycoheterotrophic orchids – in fungal partnerships, the degree of mycoheterotrophy, and plant vigor. Our findings reveal that <i>O. patens</i> plants with coralloid rhizomes predominantly associate with saprotrophic Psathyrellaceae fungi, whereas those without coralloid rhizomes also partner with rhizoctonias and other potentially orchid mycorrhizal fungi. Notably, plants with coralloid rhizomes exhibited enriched <sup>13</sup>C signatures, indicating a greater reliance on fungal-derived carbon. These plants also demonstrated more vigorous flowering scapes and produced a higher number of flowers, suggesting that mycoheterotrophy significantly enhances plant vigor. This study provides rare insights into the adaptive significance of mycoheterotrophy. Recent research suggests that some partially mycoheterotrophic orchids can adjust their heterotrophic status to optimize carbon resource use under specific conditions, such as low-light environments. However, an increased proportion of fungal-derived carbon may sometimes merely reflect reduced photosynthesis in such conditions, thereby amplifying the apparent contribution of fungal-derived carbon. Our findings offer more direct evidence that carbon acquisition via mycoheterotrophy is beneficial for partially mycoheterotrophic orchids.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.70045","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Genome editing, an opportunity to revive soybean cultivation in Europe
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.17266
Manon Monfort, Julia Buitink, Frank Roeber, Fabien Nogué

Soybean (Glycine max Merr.) is the world's most important oilseed crop and its ability to fix atmospheric nitrogen makes it a cornerstone of sustainable agriculture. Despite its importance, Europe relies heavily on imports, leading to environmental and economic vulnerabilities. To address these challenges, the European Union has implemented policies to boost local soybean production, emphasizing sustainable practices and reduced dependency on imports. However, conventional breeding methods are time-consuming and may not keep pace with the rapid environmental and consumer habit changes. Genome-editing technologies, such as CRISPR-Cas, offer precise and efficient tools for developing soybean varieties tailored to European conditions. These technologies can enhance traits related to precocity, stress responses, yield and quality that are essential for adapting to climate change and promoting ecological sustainability. This review explores the integration of genome editing (GE) in soybean breeding, highlighting its potential in advancing the agroecological transition in Europe. By having a clear regulation and enhancing breeding efforts, GE can significantly contribute to developing resilient and sustainable soybean varieties, fostering a competitive and environmentally friendly European agriculture.

{"title":"Genome editing, an opportunity to revive soybean cultivation in Europe","authors":"Manon Monfort,&nbsp;Julia Buitink,&nbsp;Frank Roeber,&nbsp;Fabien Nogué","doi":"10.1111/tpj.17266","DOIUrl":"https://doi.org/10.1111/tpj.17266","url":null,"abstract":"<p>Soybean (<i>Glycine max</i> Merr.) is the world's most important oilseed crop and its ability to fix atmospheric nitrogen makes it a cornerstone of sustainable agriculture. Despite its importance, Europe relies heavily on imports, leading to environmental and economic vulnerabilities. To address these challenges, the European Union has implemented policies to boost local soybean production, emphasizing sustainable practices and reduced dependency on imports. However, conventional breeding methods are time-consuming and may not keep pace with the rapid environmental and consumer habit changes. Genome-editing technologies, such as CRISPR-Cas, offer precise and efficient tools for developing soybean varieties tailored to European conditions. These technologies can enhance traits related to precocity, stress responses, yield and quality that are essential for adapting to climate change and promoting ecological sustainability. This review explores the integration of genome editing (GE) in soybean breeding, highlighting its potential in advancing the agroecological transition in Europe. By having a clear regulation and enhancing breeding efforts, GE can significantly contribute to developing resilient and sustainable soybean varieties, fostering a competitive and environmentally friendly European agriculture.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17266","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439067","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A cell fractionation and quantitative proteomics pipeline to enable functional analyses of cotton fiber development
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.17246
Youngwoo Lee, Heena Rani, Eileen L. Mallery, Daniel B. Szymanski

Cotton fibers are aerial trichoblasts that employ a highly polarized diffuse growth mechanism to emerge from the developing ovule epidermis. After executing a complicated morphogenetic program, the cells reach lengths over 2 cm and serve as the foundation of a multi-billion-dollar textile industry. Important traits such as fiber diameter, length, and strength are defined by the growth patterns and cell wall properties of individual cells. At present, the ability to engineer fiber traits is limited by our lack of understanding regarding the primary controls governing the rate, duration, and patterns of cell growth. To gain insights into the compartmentalized functions of proteins in cotton fiber cells, we developed a label-free liquid chromatography mass spectrometry method for systems-level analyses of fiber proteome. Purified fibers from a single locule were used to fractionate the fiber proteome into apoplast (APOT), membrane-associated (p200), and crude cytosolic (s200) fractions. Subsequently, proteins were identified, and their localizations and potential functions were analyzed using combinations of size exclusion chromatography, statistical and bioinformatic analyses. This method had good coverage of the p200 and APOT fractions, the latter of which was dominated by proteins associated with particulate membrane-enclosed compartments. The apoplastic proteome was diverse, the proteins were not degraded, and some displayed distinct multimerization states compared to their cytosolic pool. This quantitative proteomic pipeline can be used to improve coverage and functional analyses of the cotton fiber proteome as a function of developmental time or differing genotypes.

{"title":"A cell fractionation and quantitative proteomics pipeline to enable functional analyses of cotton fiber development","authors":"Youngwoo Lee,&nbsp;Heena Rani,&nbsp;Eileen L. Mallery,&nbsp;Daniel B. Szymanski","doi":"10.1111/tpj.17246","DOIUrl":"https://doi.org/10.1111/tpj.17246","url":null,"abstract":"<p>Cotton fibers are aerial trichoblasts that employ a highly polarized diffuse growth mechanism to emerge from the developing ovule epidermis. After executing a complicated morphogenetic program, the cells reach lengths over 2 cm and serve as the foundation of a multi-billion-dollar textile industry. Important traits such as fiber diameter, length, and strength are defined by the growth patterns and cell wall properties of individual cells. At present, the ability to engineer fiber traits is limited by our lack of understanding regarding the primary controls governing the rate, duration, and patterns of cell growth. To gain insights into the compartmentalized functions of proteins in cotton fiber cells, we developed a label-free liquid chromatography mass spectrometry method for systems-level analyses of fiber proteome. Purified fibers from a single locule were used to fractionate the fiber proteome into apoplast (APO<sub>T</sub>), membrane-associated (p200), and crude cytosolic (s200) fractions. Subsequently, proteins were identified, and their localizations and potential functions were analyzed using combinations of size exclusion chromatography, statistical and bioinformatic analyses. This method had good coverage of the p200 and APO<sub>T</sub> fractions, the latter of which was dominated by proteins associated with particulate membrane-enclosed compartments. The apoplastic proteome was diverse, the proteins were not degraded, and some displayed distinct multimerization states compared to their cytosolic pool. This quantitative proteomic pipeline can be used to improve coverage and functional analyses of the cotton fiber proteome as a function of developmental time or differing genotypes.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.17246","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Ecological divergence of sibling allopolyploid marsh orchids is associated with species specific plasticity and distinct fungal communities
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.70001
Katie Emelianova, Anna-Sophie Hawranek, Mimmi C. Eriksson, Thomas M. Wolfe, Ovidiu Paun

Phenotypic plasticity, the dynamic adjustment of traits to environmental variations, is crucial for enabling species to exploit broader niches and withstand suboptimal conditions. This adaptability is particularly relevant for newly formed allopolyploids, which possess redundant gene copies and must become established in diverse environments distinct from their parents and other relatives. By evaluating gene expression and root mycobiome among two ecologically divergent sibling allopolyploid marsh orchids (Dactylorhiza majalis and D. traunsteineri) in reciprocal transplants at localities where both species are native, we aimed to understand the drivers of species persistence in the face of interspecific gene flow. Despite consistent abiotic differences characterising the alternative environments at each locality, the majority of gene expression differences between the allopolyploids appears to be plastic. Ecologically relevant processes, such as photosynthesis and transmembrane transport, include some genes that are differentially expressed between the two orchids regardless of the environment, while others change their activity plastically in one species or the other. This suggests that although plasticity helps define the specific ecological range of each sibling allopolyploid, it also mediates gene flow between them, thereby preventing differentiation. Extending our investigations to the root mycobiome, we uncover more diverse fungal communities for either species when grown in the environment with nutrient-poor soils, indicating that both abiotic and biotic factors drive the distribution of sibling marsh orchids. Altogether, our results indicate that plasticity can simultaneously promote diversification and homogenisation of lineages, influencing the establishment and persistence of recurrently formed allopolyploid species.

{"title":"Ecological divergence of sibling allopolyploid marsh orchids is associated with species specific plasticity and distinct fungal communities","authors":"Katie Emelianova,&nbsp;Anna-Sophie Hawranek,&nbsp;Mimmi C. Eriksson,&nbsp;Thomas M. Wolfe,&nbsp;Ovidiu Paun","doi":"10.1111/tpj.70001","DOIUrl":"https://doi.org/10.1111/tpj.70001","url":null,"abstract":"<p>Phenotypic plasticity, the dynamic adjustment of traits to environmental variations, is crucial for enabling species to exploit broader niches and withstand suboptimal conditions. This adaptability is particularly relevant for newly formed allopolyploids, which possess redundant gene copies and must become established in diverse environments distinct from their parents and other relatives. By evaluating gene expression and root mycobiome among two ecologically divergent sibling allopolyploid marsh orchids (<i>Dactylorhiza majalis</i> and <i>D. traunsteineri</i>) in reciprocal transplants at localities where both species are native, we aimed to understand the drivers of species persistence in the face of interspecific gene flow. Despite consistent abiotic differences characterising the alternative environments at each locality, the majority of gene expression differences between the allopolyploids appears to be plastic. Ecologically relevant processes, such as photosynthesis and transmembrane transport, include some genes that are differentially expressed between the two orchids regardless of the environment, while others change their activity plastically in one species or the other. This suggests that although plasticity helps define the specific ecological range of each sibling allopolyploid, it also mediates gene flow between them, thereby preventing differentiation. Extending our investigations to the root mycobiome, we uncover more diverse fungal communities for either species when grown in the environment with nutrient-poor soils, indicating that both abiotic and biotic factors drive the distribution of sibling marsh orchids. Altogether, our results indicate that plasticity can simultaneously promote diversification and homogenisation of lineages, influencing the establishment and persistence of recurrently formed allopolyploid species.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.70001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Two O-acyltransferases from the diterpene biosynthetic gene cluster of Euphorbia lathyris contribute to the structural diversity of medicinal macrocyclic diterpenoid esters biosynthesis
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-19 DOI: 10.1111/tpj.70003
Wanli Zhao, Fan Wang, Pirui Li, Linwei Li, Lukas Ernst, Long Huang, Mei Tian, Wei Lv, Shu Xu, Fei Liu, Guyin Lin, Hui Lyu, Christian Paetz, Xu Feng, Yu Chen

Macrocyclic diterpenoid esters from Euphorbiaceae plants hold significant medicinal value owing to their structural diversity and for attributing structural uniqueness and biological efficacy. However, the responsible enzymes for the acylation of macrocyclic diterpenoids remain unknown. We identified two macrocyclic diterpenoid O-acyltransferases, ElBAHD16 and ElBAHD35, from the diterpene biosynthetic gene cluster of Euphorbia lathyris. ElBAHD16 and ElBAHD35 were characterized both in vitro (using Escherichia coli) and in vivo (using Nicotiana benthamiana and E. lathyris) and exhibited mono-acylation activities toward the hydroxy groups of their substrates, 7-hydroxylathyrol and lathyrol. ElBAHD16 showed not only regioselectivity toward the 7-OH group of 7-hydroxylathyrol but also donor promiscuity, thereby producing three different mono-acylation products. Conversely, ElBAHD35 demonstrated specific recognition for the 5-OH group of 7-hydroxylathyrol and lathyrol, thereby mediating mono-acetylation reactions with acetyl-CoA, showing donor specificity. Site-directed mutagenesis revealed that residues H154 and T363 in ElBAHD16 are critical for its catalytic activity. Notably, the Q35 residue enhanced the efficiency of ElBAHD16, while the M296, N292, and F394 residues were crucial for its donor promiscuity. These findings elucidate the last step in the biosynthesis of macrocyclic diterpenoid esters and highlight the contribution of acyltransferases to the structural diversity of diterpenoids.

{"title":"Two O-acyltransferases from the diterpene biosynthetic gene cluster of Euphorbia lathyris contribute to the structural diversity of medicinal macrocyclic diterpenoid esters biosynthesis","authors":"Wanli Zhao,&nbsp;Fan Wang,&nbsp;Pirui Li,&nbsp;Linwei Li,&nbsp;Lukas Ernst,&nbsp;Long Huang,&nbsp;Mei Tian,&nbsp;Wei Lv,&nbsp;Shu Xu,&nbsp;Fei Liu,&nbsp;Guyin Lin,&nbsp;Hui Lyu,&nbsp;Christian Paetz,&nbsp;Xu Feng,&nbsp;Yu Chen","doi":"10.1111/tpj.70003","DOIUrl":"https://doi.org/10.1111/tpj.70003","url":null,"abstract":"<div>\u0000 \u0000 <p>Macrocyclic diterpenoid esters from Euphorbiaceae plants hold significant medicinal value owing to their structural diversity and for attributing structural uniqueness and biological efficacy. However, the responsible enzymes for the acylation of macrocyclic diterpenoids remain unknown. We identified two macrocyclic diterpenoid <i>O</i>-acyltransferases, ElBAHD16 and ElBAHD35, from the diterpene biosynthetic gene cluster of <i>Euphorbia lathyris</i>. ElBAHD16 and ElBAHD35 were characterized both <i>in vitro</i> (using <i>Escherichia coli</i>) and <i>in vivo</i> (using <i>Nicotiana benthamiana</i> and <i>E. lathyris</i>) and exhibited mono-acylation activities toward the hydroxy groups of their substrates, 7-hydroxylathyrol and lathyrol. ElBAHD16 showed not only regioselectivity toward the 7-OH group of 7-hydroxylathyrol but also donor promiscuity, thereby producing three different mono-acylation products. Conversely, ElBAHD35 demonstrated specific recognition for the 5-OH group of 7-hydroxylathyrol and lathyrol, thereby mediating mono-acetylation reactions with acetyl-CoA, showing donor specificity. Site-directed mutagenesis revealed that residues H154 and T363 in ElBAHD16 are critical for its catalytic activity. Notably, the Q35 residue enhanced the efficiency of ElBAHD16, while the M296, N292, and F394 residues were crucial for its donor promiscuity. These findings elucidate the last step in the biosynthesis of macrocyclic diterpenoid esters and highlight the contribution of acyltransferases to the structural diversity of diterpenoids.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143439071","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Hydrogen sulfide interferes with ethylene biosynthesis and signaling pathway in tomato by the mediation of SlERF.D2 persulfidation
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-17 DOI: 10.1111/tpj.70000
Ying Sun, Min Zhang, Meihui Geng, Zhikun Geng, Zixu Lu, Nannan Liu, Zhanmin Liu, Dexin Zeng, Gaifang Yao, Kangdi Hu, Hua Zhang

Hydrogen sulfide (H2S), as a signaling molecule, is found to delay fruit ripening and senescence by antagonizing the biosynthesis and signaling of ethylene, whereas the mechanism remains unclear. In the current work, exogenous H2S fumigation could alleviate tomato fruit ripening and an ethylene response factor SlERF.D2 was found to be persulfidated at Cys35 by mass spectrometry analysis. Meanwhile, ethylene biosynthesis related genes SlACS1 and SlACO3 were significantly downregulated at gene expression level in H2S-treated fruit. By CRISPR/Cas9 and gene overexpression, we showed that overexpression of SlERF.D2 promoted fruit ripening by accelerating chlorophyll degradation and carotenoid accumulation and upregulating the expression of ripening related genes SlPAO, SlPPH, SlSGR1, SlACS1, SlACS2, SlACS4, SlEIN2, SlACO1, and SlACO3, while the mutation of slerf.d2 delayed fruit ripening. Additionally, slerf.d2 mutant showed delayed ethylene production during tomato fruit ripening. Moreover, SlERF.D2 was found to interact with the kinase SlMAPK4 and was phosphorylated at Ser42 by yeast two-hybrid screening, pull down and LC–MS/MS. By cis-element analysis, electrophoretic mobility shift assay and dual-luciferase assay, SlERF.D2 could activate the transcription of the ethylene pathway-associated gene SlACO3 and SlEIN2. Besides, we provided evidence that SlERF.D2 persulfidation weakened the transcriptional activity of SlERF.D2 on the target gene SlACO3 and SlEIN2. In contrast, SlMAPK4-mediated phosphorylation enhanced SlERF.D2's transcriptional activation activity on SlACO3 and SlEIN2. Therefore, the present research provides insights into the mechanism of H2S in antagonizing the biosynthesis and signaling transduction of ethylene and reveals the importance of SlERF.D2 persulfidation and phosphorylation in dynamically regulating tomato fruit ripening.

{"title":"Hydrogen sulfide interferes with ethylene biosynthesis and signaling pathway in tomato by the mediation of SlERF.D2 persulfidation","authors":"Ying Sun,&nbsp;Min Zhang,&nbsp;Meihui Geng,&nbsp;Zhikun Geng,&nbsp;Zixu Lu,&nbsp;Nannan Liu,&nbsp;Zhanmin Liu,&nbsp;Dexin Zeng,&nbsp;Gaifang Yao,&nbsp;Kangdi Hu,&nbsp;Hua Zhang","doi":"10.1111/tpj.70000","DOIUrl":"https://doi.org/10.1111/tpj.70000","url":null,"abstract":"<div>\u0000 \u0000 <p>Hydrogen sulfide (H<sub>2</sub>S), as a signaling molecule, is found to delay fruit ripening and senescence by antagonizing the biosynthesis and signaling of ethylene, whereas the mechanism remains unclear. In the current work, exogenous H<sub>2</sub>S fumigation could alleviate tomato fruit ripening and an ethylene response factor SlERF.D2 was found to be persulfidated at Cys35 by mass spectrometry analysis. Meanwhile, ethylene biosynthesis related genes <i>SlACS1</i> and <i>SlACO3</i> were significantly downregulated at gene expression level in H<sub>2</sub>S-treated fruit. By CRISPR/Cas9 and gene overexpression, we showed that overexpression of <i>SlERF.D2</i> promoted fruit ripening by accelerating chlorophyll degradation and carotenoid accumulation and upregulating the expression of ripening related genes <i>SlPAO</i>, <i>SlPPH</i>, <i>SlSGR1</i>, <i>SlACS1</i>, <i>SlACS2</i>, <i>SlACS4</i>, <i>SlEIN2</i>, <i>SlACO1</i>, and <i>SlACO3</i>, while the mutation of <i>slerf.d2</i> delayed fruit ripening. Additionally, <i>slerf.d2</i> mutant showed delayed ethylene production during tomato fruit ripening. Moreover, SlERF.D2 was found to interact with the kinase SlMAPK4 and was phosphorylated at Ser42 by yeast two-hybrid screening, pull down and LC–MS/MS. By cis-element analysis, electrophoretic mobility shift assay and dual-luciferase assay, SlERF.D2 could activate the transcription of the ethylene pathway-associated gene <i>SlACO3</i> and <i>SlEIN2</i>. Besides, we provided evidence that SlERF.D2 persulfidation weakened the transcriptional activity of SlERF.D2 on the target gene <i>SlACO3</i> and <i>SlEIN2</i>. In contrast, SlMAPK4-mediated phosphorylation enhanced SlERF.D2's transcriptional activation activity on <i>SlACO3</i> and <i>SlEIN2</i>. Therefore, the present research provides insights into the mechanism of H<sub>2</sub>S in antagonizing the biosynthesis and signaling transduction of ethylene and reveals the importance of SlERF.D2 persulfidation and phosphorylation in dynamically regulating tomato fruit ripening.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143431506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Molecular chaperon HSP70-9 is required for clathrin-mediated endocytosis in Arabidopsis
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-17 DOI: 10.1111/tpj.17265
Tingting Shen, Zimeng Liu, Jiaoteng Bai, Xiaoting Zhai, Shanshan Wei, Wenyan Xu, Ning Jia, Bing Li

Clathrin-mediated endocytosis (CME) plays a key role in the internalization of plasma membrane-localized proteins, lipids and extracellular substances; however, the regulatory mechanism of CME in plants is unclear. In this work, we demonstrated that molecular chaperon HSP70-9 is required for CME in Arabidopsis thaliana. Knocking out the HSP70-9 gene led to auxin-related phenotypes and the accumulation of auxin efflux carrier PINs in root cells, indicating that HSP70-9 is involved in the PIN trafficking process. Immunolocalization studies showed that the HSP70-9 was localized in mitochondria, vesicles and cytoplasm. Next, we found that HSP70-9 and clathrin light chain 1 (CLC1) were co-localized in cells, and there was a strong interaction between the two. HSP70-9 knockout led to an increase in CLC1-related vesicle number in root cells, and the application of protein synthesis inhibitor cycloheximide did not significantly inhibit the increase in the hsp70-9a mutant. HSP70-9 knockout did not significantly affect the level of CLC1 mRNA, but reduced the abundance of free CLC1 protein in root cells, indicating that HSP70-9 might be involved in the stability of CLC1-related vesicles. Moreover, our data indicated that the introduction of the exogenous CLC1 gene rescued the hsp70-9a seedlings likely through promoting PIN trafficking. Furthermore, the role of HSP70-9 in the depolymerization of clathrin-coated vesicles (CCVs) was investigated in vitro. The result indicated that the addition of HSP70-9 promoted the release of CLC1 from the CCVs. Taken together, our data suggest that HSP70-9 affects PIN trafficking likely through facilitating the release of CLC1 from the CCVs.

{"title":"Molecular chaperon HSP70-9 is required for clathrin-mediated endocytosis in Arabidopsis","authors":"Tingting Shen,&nbsp;Zimeng Liu,&nbsp;Jiaoteng Bai,&nbsp;Xiaoting Zhai,&nbsp;Shanshan Wei,&nbsp;Wenyan Xu,&nbsp;Ning Jia,&nbsp;Bing Li","doi":"10.1111/tpj.17265","DOIUrl":"https://doi.org/10.1111/tpj.17265","url":null,"abstract":"<div>\u0000 \u0000 <p>Clathrin-mediated endocytosis (CME) plays a key role in the internalization of plasma membrane-localized proteins, lipids and extracellular substances; however, the regulatory mechanism of CME in plants is unclear. In this work, we demonstrated that molecular chaperon HSP70-9 is required for CME in <i>Arabidopsis thaliana</i>. Knocking out the <i>HSP70-9</i> gene led to auxin-related phenotypes and the accumulation of auxin efflux carrier PINs in root cells, indicating that HSP70-9 is involved in the PIN trafficking process. Immunolocalization studies showed that the HSP70-9 was localized in mitochondria, vesicles and cytoplasm. Next, we found that HSP70-9 and clathrin light chain 1 (CLC1) were co-localized in cells, and there was a strong interaction between the two. <i>HSP70-9</i> knockout led to an increase in CLC1-related vesicle number in root cells, and the application of protein synthesis inhibitor cycloheximide did not significantly inhibit the increase in the <i>hsp70-9a</i> mutant. <i>HSP70-9</i> knockout did not significantly affect the level of <i>CLC1</i> mRNA, but reduced the abundance of free CLC1 protein in root cells, indicating that HSP70-9 might be involved in the stability of CLC1-related vesicles. Moreover, our data indicated that the introduction of the exogenous <i>CLC1</i> gene rescued the <i>hsp70-9a</i> seedlings likely through promoting PIN trafficking. Furthermore, the role of HSP70-9 in the depolymerization of clathrin-coated vesicles (CCVs) was investigated <i>in vitro</i>. The result indicated that the addition of HSP70-9 promoted the release of CLC1 from the CCVs. Taken together, our data suggest that HSP70-9 affects PIN trafficking likely through facilitating the release of CLC1 from the CCVs.</p>\u0000 </div>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143431505","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Chromosome engineering points to the cis-acting mechanism of chromosome arm-specific telomere length setting and robustness of plant phenotype, chromatin structure and gene expression
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-02-17 DOI: 10.1111/tpj.70024
Ondřej Helia, Barbora Matúšová, Kateřina Havlová, Anna Hýsková, Martin Lyčka, Natalja Beying, Holger Puchta, Jiří Fajkus, Miloslava Fojtová

The study investigates the impact of targeted chromosome engineering on telomere dynamics, chromatin structure, gene expression, and phenotypic stability in Arabidopsis thaliana. Using precise CRISPR/Cas-based engineering, reciprocal translocations of chromosome arms were introduced between non-homologous chromosomes. The subsequent homozygous generations of plants were assessed for phenotype, transcriptomic changes and chromatin modifications near translocation breakpoints, and telomere length maintenance. Phenotypically, translocated lines were indistinguishable from wild-type plants, as confirmed through morphological assessments and principal component analysis. Gene expression profiling detected minimal differential expression, with affected genes dispersed across the genome, indicating negligible transcriptional impact. Similarly, ChIPseq analysis showed no substantial alterations in the enrichment of key histone marks (H3K27me3, H3K4me1, H3K56ac) near junction sites or across the genome. Finally, bulk and arm-specific telomere lengths remained stable across multiple generations, except for minor variations in one translocation line. These findings highlight the remarkable genomic and phenotypic robustness of A. thaliana despite large-scale chromosomal rearrangements. The study offers insights into the cis-acting mechanisms underlying chromosome arm-specific telomere length setting and establishes the feasibility of chromosome engineering for studies of plant genome evolution and crop improvement strategies.

{"title":"Chromosome engineering points to the cis-acting mechanism of chromosome arm-specific telomere length setting and robustness of plant phenotype, chromatin structure and gene expression","authors":"Ondřej Helia,&nbsp;Barbora Matúšová,&nbsp;Kateřina Havlová,&nbsp;Anna Hýsková,&nbsp;Martin Lyčka,&nbsp;Natalja Beying,&nbsp;Holger Puchta,&nbsp;Jiří Fajkus,&nbsp;Miloslava Fojtová","doi":"10.1111/tpj.70024","DOIUrl":"https://doi.org/10.1111/tpj.70024","url":null,"abstract":"<p>The study investigates the impact of targeted chromosome engineering on telomere dynamics, chromatin structure, gene expression, and phenotypic stability in <i>Arabidopsis thaliana</i>. Using precise CRISPR/Cas-based engineering, reciprocal translocations of chromosome arms were introduced between non-homologous chromosomes. The subsequent homozygous generations of plants were assessed for phenotype, transcriptomic changes and chromatin modifications near translocation breakpoints, and telomere length maintenance. Phenotypically, translocated lines were indistinguishable from wild-type plants, as confirmed through morphological assessments and principal component analysis. Gene expression profiling detected minimal differential expression, with affected genes dispersed across the genome, indicating negligible transcriptional impact. Similarly, ChIPseq analysis showed no substantial alterations in the enrichment of key histone marks (H3K27me3, H3K4me1, H3K56ac) near junction sites or across the genome. Finally, bulk and arm-specific telomere lengths remained stable across multiple generations, except for minor variations in one translocation line. These findings highlight the remarkable genomic and phenotypic robustness of <i>A. thaliana</i> despite large-scale chromosomal rearrangements. The study offers insights into the <i>cis</i>-acting mechanisms underlying chromosome arm-specific telomere length setting and establishes the feasibility of chromosome engineering for studies of plant genome evolution and crop improvement strategies.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"121 4","pages":""},"PeriodicalIF":6.2,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.70024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143431502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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The Plant Journal
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