Jialing Zhang, Li Chen, Weiwei Yao, Yupeng Cai, Wensheng Hou
Flowering time and drought resistance are two pivotal agronomic traits in soybean. Elucidating coregulatory modules that link soybean flowering and drought response is essential for constructing comprehensive molecular maps of trait coupling. In this study, we identified that MORN-MOTIF REPEAT PROTEIN REGULATING FLOWERING LIKE (GmMRFL) gene functions as a bifunctional regulator that concurrently promotes floral transition by upregulating the expression of Flowering Locus T (FT) genes and enhances drought resilience through stomatal adjustment, accompanied by abscisic acid (ABA) signaling and reactive oxygen species (ROS) suppression. In addition, the transcription factor AP2/ETHYLENE-RESPONSIVE FACTOR 011 (GmERF011) specifically binds to and activates the Hap1 promoter variant of GmMRFL, thereby promoting the upregulation of GmMRFL expression. Phenotypic analyses of hairy roots validated the role of GmERF011 in enhancing drought tolerance in soybean. Integrated molecular analyses revealed that GmMRFL interacts with ANKYRIN REPEAT DOMAIN PROTEIN 2 (GmANK2). These findings demonstrate that GmMRFL serves as a molecular hub that coordinately modulates photoperiod-dependent flowering regulation and drought adaptation, thereby establishing it as a prime target for multi-trait engineering in precision crop breeding.
开花时间和抗旱性是大豆的两个关键农艺性状。阐明大豆开花与干旱响应之间的共调控模块，是构建全面的性状偶联分子图谱的基础。本研究发现，MORN-MOTIF REPEAT PROTEIN REGULATING blossom LIKE （GmMRFL）基因作为双功能调控因子，通过上调开花位点T （FT）基因的表达促进开花转变，同时通过调节气孔增强抗旱性，并伴随脱落酸（ABA）信号和活性氧（ROS）抑制。此外，转录因子AP2/乙烯响应因子011 （GmERF011）特异性结合并激活GmMRFL的Hap1启动子变体，从而促进GmMRFL表达上调。毛状根表型分析证实了GmERF011对大豆抗旱性的增强作用。综合分子分析显示，GmMRFL与ANKYRIN REPEAT DOMAIN PROTEIN 2 （GmANK2）相互作用。这些发现表明，GmMRFL作为协调调节光周期依赖的开花调节和干旱适应的分子枢纽，从而使其成为作物精准育种中多性状工程的主要靶点。
{"title":"The GmERF011- GmMRFL regulatory module integrates floral transition and drought stress adaptation in soybean","authors":"Jialing Zhang, Li Chen, Weiwei Yao, Yupeng Cai, Wensheng Hou","doi":"10.1093/plphys/kiag042","DOIUrl":"https://doi.org/10.1093/plphys/kiag042","url":null,"abstract":"Flowering time and drought resistance are two pivotal agronomic traits in soybean. Elucidating coregulatory modules that link soybean flowering and drought response is essential for constructing comprehensive molecular maps of trait coupling. In this study, we identified that MORN-MOTIF REPEAT PROTEIN REGULATING FLOWERING LIKE (GmMRFL) gene functions as a bifunctional regulator that concurrently promotes floral transition by upregulating the expression of Flowering Locus T (FT) genes and enhances drought resilience through stomatal adjustment, accompanied by abscisic acid (ABA) signaling and reactive oxygen species (ROS) suppression. In addition, the transcription factor AP2/ETHYLENE-RESPONSIVE FACTOR 011 (GmERF011) specifically binds to and activates the Hap1 promoter variant of GmMRFL, thereby promoting the upregulation of GmMRFL expression. Phenotypic analyses of hairy roots validated the role of GmERF011 in enhancing drought tolerance in soybean. Integrated molecular analyses revealed that GmMRFL interacts with ANKYRIN REPEAT DOMAIN PROTEIN 2 (GmANK2). These findings demonstrate that GmMRFL serves as a molecular hub that coordinately modulates photoperiod-dependent flowering regulation and drought adaptation, thereby establishing it as a prime target for multi-trait engineering in precision crop breeding.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"44 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146110533","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}
Hiruni N Weerasooriya, Isaiah C M Pabuayon, Xiaozhuo Wang, Himanshu S Mehra, Nidhi Kulkarni, Nicholas Ferrari, David J Longstreth, James V Moroney
The physiological role of chloroplast carbonic anhydrases (CAs) has long been debated, particularly in the context of photosynthesis. While early hypotheses proposed that CAs enhance CO₂ assimilation by rapidly accessing the HCO3- pool, direct evidence has been lacking. In this study, we examined Arabidopsis (Arabidopsis thaliana) mutants lacking both chloroplast-localized βCA1 (AT3G01500) and βCA5 (AT4G33580) to assess their impact on plant growth and photosynthetic performance. Our results show that plants deficient in chloroplast CA activity are unable to grow under ambient CO₂ conditions (400 μL·L⁻¹) but can complete their life cycle under elevated CO₂ levels (≥12,000 μL·L⁻¹). However, CO₂ assimilation rates and ΦII measurements in CA-deficient plants were comparable to those in wild-type plants under 0.04% (400 μL·L⁻¹), 0.4% (4,000 μL·L⁻¹), and 4% CO₂ (40,000 μL·L⁻¹) concentrations, indicating that chloroplast CAs are not essential for photosynthetic CO₂ fixation. Instead, our findings suggest that chloroplast CA activity is critical for supporting other metabolic pathways, namely amino acid, nucleic acid, and fatty acid biosynthesis. Expression of the Chlamydomonas (Chlamydomonas reinhardtii) bicarbonate transporter LCIA in chloroplast CA mutants partially rescued the growth phenotype under near-ambient CO₂ conditions. These LCIA-complemented lines showed no difference in photosynthesis, further supporting the role of CAs in non-photosynthetic reactions. This work provides direct evidence that while chloroplast CAs are dispensable for photosynthesis, they are essential for plant growth and development under ambient CO₂ due to their role in increasing the bicarbonate concentration for specific anaplerotic pathways.
{"title":"Plastid carbonic anhydrases are essential for growth of Arabidopsis on ambient air but not for photosynthesis","authors":"Hiruni N Weerasooriya, Isaiah C M Pabuayon, Xiaozhuo Wang, Himanshu S Mehra, Nidhi Kulkarni, Nicholas Ferrari, David J Longstreth, James V Moroney","doi":"10.1093/plphys/kiag048","DOIUrl":"https://doi.org/10.1093/plphys/kiag048","url":null,"abstract":"The physiological role of chloroplast carbonic anhydrases (CAs) has long been debated, particularly in the context of photosynthesis. While early hypotheses proposed that CAs enhance CO₂ assimilation by rapidly accessing the HCO3- pool, direct evidence has been lacking. In this study, we examined Arabidopsis (Arabidopsis thaliana) mutants lacking both chloroplast-localized βCA1 (AT3G01500) and βCA5 (AT4G33580) to assess their impact on plant growth and photosynthetic performance. Our results show that plants deficient in chloroplast CA activity are unable to grow under ambient CO₂ conditions (400 μL·L⁻¹) but can complete their life cycle under elevated CO₂ levels (≥12,000 μL·L⁻¹). However, CO₂ assimilation rates and ΦII measurements in CA-deficient plants were comparable to those in wild-type plants under 0.04% (400 μL·L⁻¹), 0.4% (4,000 μL·L⁻¹), and 4% CO₂ (40,000 μL·L⁻¹) concentrations, indicating that chloroplast CAs are not essential for photosynthetic CO₂ fixation. Instead, our findings suggest that chloroplast CA activity is critical for supporting other metabolic pathways, namely amino acid, nucleic acid, and fatty acid biosynthesis. Expression of the Chlamydomonas (Chlamydomonas reinhardtii) bicarbonate transporter LCIA in chloroplast CA mutants partially rescued the growth phenotype under near-ambient CO₂ conditions. These LCIA-complemented lines showed no difference in photosynthesis, further supporting the role of CAs in non-photosynthetic reactions. This work provides direct evidence that while chloroplast CAs are dispensable for photosynthesis, they are essential for plant growth and development under ambient CO₂ due to their role in increasing the bicarbonate concentration for specific anaplerotic pathways.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"5 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146101701","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}
She Men,Fen Zhao,Xiangnan Zhang,Zeyuan Guan,Jiawen Xu,Yuan Xue,Haozhe Tan,Xiao Chang,Guannan Zhao,Chunmiao Li,Zhonghua Liu,Xianlong Zhang,Ping Yin,Lili Tu
Expansins are pivotal cell wall loosening proteins that facilitate turgor-driven extension of plant cell walls. Expansin-like A (EXLA) proteins represent a subfamily, but their interaction with polysaccharides remains poorly understood during primary cell wall growth, hindered by challenges in achieving active heterologous expression for in vitro analysis. Using an insect secretion-based expression system, we successfully expressed and purified EXLA proteins. Screening eight different polysaccharides showed that EXLAs exhibit a preference for binding to negatively charged polygalacturonic acid (PGA) and rhamnogalacturonan I (RG-I), pivotal components of pectin in the primary cell wall matrix. The crystal structure of EXLA1 was resolved at 2.5 Å resolution, revealing three crucial positively charged surfaces for pectin electrostatic interaction, and mutating these basic amino acids to alanine significantly reduced the binding ability. Moreover, recombinant EXLA1 promoted the extension of heat-inactivated cucumber hypocotyl walls under acidic conditions, indicating its intrinsic wall-loosening activity in vitro. EXLA1 overexpression resulted in a remodeled cell wall structure, suggesting EXLAs affect cell wall growth. These findings unveil EXLAs function during cell wall development by binding pectin through electrostatic interactions.
扩张蛋白是关键的细胞壁松动蛋白，促进植物细胞壁膨胀驱动的延伸。Expansin-like A （EXLA）蛋白代表一个亚家族，但在原代细胞壁生长过程中，它们与多糖的相互作用仍然知之甚少，这受到体外分析中实现活性异源表达的挑战的阻碍。利用基于昆虫分泌物的表达系统，我们成功地表达和纯化了EXLA蛋白。对8种不同多糖的筛选表明，EXLAs倾向于与带负电荷的聚半乳糖醛酸（PGA）和鼠李糖半乳糖醛酸I （RG-I）结合，后者是初代细胞壁基质中果胶的关键成分。在2.5 Å分辨率下对EXLA1的晶体结构进行了解析，揭示了果胶静电相互作用的三个关键正电荷表面，将这些碱性氨基酸突变为丙氨酸显著降低了结合能力。此外，重组EXLA1在酸性条件下促进了热失活黄瓜下胚轴壁的延伸，表明其在体外具有内在的松壁活性。EXLA1过表达导致细胞壁结构重塑，提示EXLA1影响细胞壁生长。这些发现揭示了EXLAs通过静电相互作用结合果胶在细胞壁发育过程中的作用。
{"title":"Expansin-like A binds to pectin via electrostatic forces and remodels the plant cell wall.","authors":"She Men,Fen Zhao,Xiangnan Zhang,Zeyuan Guan,Jiawen Xu,Yuan Xue,Haozhe Tan,Xiao Chang,Guannan Zhao,Chunmiao Li,Zhonghua Liu,Xianlong Zhang,Ping Yin,Lili Tu","doi":"10.1093/plphys/kiag029","DOIUrl":"https://doi.org/10.1093/plphys/kiag029","url":null,"abstract":"Expansins are pivotal cell wall loosening proteins that facilitate turgor-driven extension of plant cell walls. Expansin-like A (EXLA) proteins represent a subfamily, but their interaction with polysaccharides remains poorly understood during primary cell wall growth, hindered by challenges in achieving active heterologous expression for in vitro analysis. Using an insect secretion-based expression system, we successfully expressed and purified EXLA proteins. Screening eight different polysaccharides showed that EXLAs exhibit a preference for binding to negatively charged polygalacturonic acid (PGA) and rhamnogalacturonan I (RG-I), pivotal components of pectin in the primary cell wall matrix. The crystal structure of EXLA1 was resolved at 2.5 Å resolution, revealing three crucial positively charged surfaces for pectin electrostatic interaction, and mutating these basic amino acids to alanine significantly reduced the binding ability. Moreover, recombinant EXLA1 promoted the extension of heat-inactivated cucumber hypocotyl walls under acidic conditions, indicating its intrinsic wall-loosening activity in vitro. EXLA1 overexpression resulted in a remodeled cell wall structure, suggesting EXLAs affect cell wall growth. These findings unveil EXLAs function during cell wall development by binding pectin through electrostatic interactions.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"261 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146073127","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}
Amid global climate change, extreme high-temperature events have become increasingly frequent, posing significant threats to ecosystems and agricultural production. NAM, ATAF, and CUC transcription factors (NAC TFs) play a key role in responding to environmental stresses such as drought, salt stress, and temperature changes. This study demonstrates that SlNAC72, a member of the NAC transcription factor family, directly targets and regulates LIPOXYGENASE 10 (SlLOX10), ALLENE OXIDE SYNTHASE 1 (SlAOS1), and ALLENE OXIDE CYCLASE (SlAOC) in tomato (Solanum lycopersicum), inhibiting jasmonic acid (JA) biosynthesis and thereby negatively regulating heat tolerance. Overexpression of SlAOS1 notably increased JA accumulation under high-temperature stress and mitigated excessive reactive oxygen species (ROS) accumulation, whereas slaos1 knockout resulted in the opposite phenotype, indicating the positive role of SlAOS1 in JA biosynthesis and high-temperature stress tolerance. Further investigations revealed that SlNAC72 interacts with the E3 ubiquitin ligase MYB30-INTERACTING E3 LIGASE 1 (SlMIEL1) and that SlMIEL1 promotes JA accumulation by mediating the ubiquitination and degradation of SlNAC72, ultimately enhancing high-temperature tolerance in tomato. Additionally, as a key transcription factor in the JA signaling pathway, MYELOCYTOMATOSIS 2 (SlMYC2) directly bound to SlNAC72 and suppressed its expression. This study uncovers the central role of the SlMYC2-SlNAC72-SlMIEL1 module in regulating JA biosynthesis and elucidates how this module contributes to the molecular mechanisms underlying tomato's response to high-temperature stress via regulating JA accumulation.
{"title":"The SlMYC2-SlNAC72-SlMIEL1 module contributes to high-temperature tolerance in tomato by regulating jasmonic acid biosynthesis.","authors":"Xiangguang Meng,Zhen Kang,Guo Chen,Yue Feng,Yong Zhang,Guobin Li,Songshen Hu,Changan Zhu,Tianlai Li,Xiaohui Hu","doi":"10.1093/plphys/kiag028","DOIUrl":"https://doi.org/10.1093/plphys/kiag028","url":null,"abstract":"Amid global climate change, extreme high-temperature events have become increasingly frequent, posing significant threats to ecosystems and agricultural production. NAM, ATAF, and CUC transcription factors (NAC TFs) play a key role in responding to environmental stresses such as drought, salt stress, and temperature changes. This study demonstrates that SlNAC72, a member of the NAC transcription factor family, directly targets and regulates LIPOXYGENASE 10 (SlLOX10), ALLENE OXIDE SYNTHASE 1 (SlAOS1), and ALLENE OXIDE CYCLASE (SlAOC) in tomato (Solanum lycopersicum), inhibiting jasmonic acid (JA) biosynthesis and thereby negatively regulating heat tolerance. Overexpression of SlAOS1 notably increased JA accumulation under high-temperature stress and mitigated excessive reactive oxygen species (ROS) accumulation, whereas slaos1 knockout resulted in the opposite phenotype, indicating the positive role of SlAOS1 in JA biosynthesis and high-temperature stress tolerance. Further investigations revealed that SlNAC72 interacts with the E3 ubiquitin ligase MYB30-INTERACTING E3 LIGASE 1 (SlMIEL1) and that SlMIEL1 promotes JA accumulation by mediating the ubiquitination and degradation of SlNAC72, ultimately enhancing high-temperature tolerance in tomato. Additionally, as a key transcription factor in the JA signaling pathway, MYELOCYTOMATOSIS 2 (SlMYC2) directly bound to SlNAC72 and suppressed its expression. This study uncovers the central role of the SlMYC2-SlNAC72-SlMIEL1 module in regulating JA biosynthesis and elucidates how this module contributes to the molecular mechanisms underlying tomato's response to high-temperature stress via regulating JA accumulation.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"65 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146070012","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}
Xin Liu,Hao Yang,Siqi Liu,Qi Zhang,Jian Lin,Jiao Zhan,Li Yuan,Mingkun Yang,Feng Ge
Lysine acetylation (Kac) is a critical post-translational modification that regulates photosynthesis and carbon metabolism in cyanobacteria. However, the diversity and functional roles of lysine acetyltransferases (KATs) beyond the well-characterized cGNAT2 remain poorly defined. This study identifies and functionally characterizes the previously unannotated protein A0096, designated here as cKAT, demonstrating its capacity to catalyze Kac both in vivo and in vitro in Synechococcus sp. PCC 7002 (Synechococcus). Deletion of cKAT significantly impaired cellular growth and photosynthetic efficiency in Synechococcus. Utilizing label-free quantitative (LFQ) acetylome profiling, we identified 171 endogenous Kac sites across 137 proteins targeted by cKAT. These target proteins participate in diverse metabolic and photosynthetic pathways, indicating a broad regulatory role for cKAT in cellular physiology. Notably, ChpX, a key component of CO₂-concentrating mechanisms, was prominently acetylated by cKAT both in vivo and in vitro. We further established that cKAT specifically mediates acetylation at residue K88 of ChpX, a modification that directly modulates CO₂ uptake efficiency. This regulatory mechanism consequently influences photosynthetic performance and cellular growth in Synechococcus. Collectively, these findings establish cKAT as a central regulator of cyanobacterial carbon fixation. This work expands the known repertoire of photosynthetic acetyltransferases and provides mechanistic insights into the Kac-dependent regulation of photosynthetic processes.
{"title":"cKAT acetylation of the CO₂ hydration protein ChpX regulates the CO₂ concentrating mechanism in cyanobacteria.","authors":"Xin Liu,Hao Yang,Siqi Liu,Qi Zhang,Jian Lin,Jiao Zhan,Li Yuan,Mingkun Yang,Feng Ge","doi":"10.1093/plphys/kiag033","DOIUrl":"https://doi.org/10.1093/plphys/kiag033","url":null,"abstract":"Lysine acetylation (Kac) is a critical post-translational modification that regulates photosynthesis and carbon metabolism in cyanobacteria. However, the diversity and functional roles of lysine acetyltransferases (KATs) beyond the well-characterized cGNAT2 remain poorly defined. This study identifies and functionally characterizes the previously unannotated protein A0096, designated here as cKAT, demonstrating its capacity to catalyze Kac both in vivo and in vitro in Synechococcus sp. PCC 7002 (Synechococcus). Deletion of cKAT significantly impaired cellular growth and photosynthetic efficiency in Synechococcus. Utilizing label-free quantitative (LFQ) acetylome profiling, we identified 171 endogenous Kac sites across 137 proteins targeted by cKAT. These target proteins participate in diverse metabolic and photosynthetic pathways, indicating a broad regulatory role for cKAT in cellular physiology. Notably, ChpX, a key component of CO₂-concentrating mechanisms, was prominently acetylated by cKAT both in vivo and in vitro. We further established that cKAT specifically mediates acetylation at residue K88 of ChpX, a modification that directly modulates CO₂ uptake efficiency. This regulatory mechanism consequently influences photosynthetic performance and cellular growth in Synechococcus. Collectively, these findings establish cKAT as a central regulator of cyanobacterial carbon fixation. This work expands the known repertoire of photosynthetic acetyltransferases and provides mechanistic insights into the Kac-dependent regulation of photosynthetic processes.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"296 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146069994","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}
The shoot apical meristem (SAM) determines plant architecture, but the key components of its regulatory network remain elusive in rapeseed (Brassica napus L.). Here, we integrated transcriptomic profiling of three multilocular silique mutants (Bnaclv1, Bnaclv2, Bnaclv3) across key SAM development stages (IM, stage6 and stage8) with large-scale CRISPR/Cas9 functional screening to identify regulators of SAM maintenance. Differential gene expression and GO enrichment highlighted genes significantly associated with meristem development processes. Weighted Gene Co-expression Network Analysis (WGCNA) of stage-specific transcriptomes identified 42 candidate genes potentially related to SAM development. To enable systematic functional screening, we established a high-throughput multiplex CRISPR/Cas9 pipeline, simultaneously targeting 198 sites across 42 candidate genes through optimized sgRNA design and pooled transformation. We successfully obtained mutants for 25 genes with homozygous mutants for 9 genes. Phenotypic analysis demonstrated that mutants of BnaSCPL family genes (SCPL29, SCPL44, SCPL45) exhibited a multi-stem phenotype and disrupted SAM organization. Mechanistic studies revealed that BnaSCPL mutations disrupt the canonical CLV3/WUS feedback loop, uncovering their roles in SAM homeostasis. Additionally, knockout of BnaLFY homologs caused permanent vegetative state and sterility, demonstrating their conserved role in floral meristem identity in Brassica napus. Collectively, our study not only elucidates the critical function of BnaSCPLs in SAM maintenance but also establishes a regulatory framework for understanding meristem phase transitions in B. napus, providing potential targets for crop architecture improvement.
{"title":"Integrating transcriptomics and high-throughput gene editing uncovers shoot apical meristem regulators in Brassica napus.","authors":"Kaidi Yu,Huailin Li,Yuzhe Hu,Yalun Yu,Songyue Deng,Yang Yang,Mixia Guo,Mengting Li,Meiling Zhe,Hanzi He,Chuchuan Fan","doi":"10.1093/plphys/kiag032","DOIUrl":"https://doi.org/10.1093/plphys/kiag032","url":null,"abstract":"The shoot apical meristem (SAM) determines plant architecture, but the key components of its regulatory network remain elusive in rapeseed (Brassica napus L.). Here, we integrated transcriptomic profiling of three multilocular silique mutants (Bnaclv1, Bnaclv2, Bnaclv3) across key SAM development stages (IM, stage6 and stage8) with large-scale CRISPR/Cas9 functional screening to identify regulators of SAM maintenance. Differential gene expression and GO enrichment highlighted genes significantly associated with meristem development processes. Weighted Gene Co-expression Network Analysis (WGCNA) of stage-specific transcriptomes identified 42 candidate genes potentially related to SAM development. To enable systematic functional screening, we established a high-throughput multiplex CRISPR/Cas9 pipeline, simultaneously targeting 198 sites across 42 candidate genes through optimized sgRNA design and pooled transformation. We successfully obtained mutants for 25 genes with homozygous mutants for 9 genes. Phenotypic analysis demonstrated that mutants of BnaSCPL family genes (SCPL29, SCPL44, SCPL45) exhibited a multi-stem phenotype and disrupted SAM organization. Mechanistic studies revealed that BnaSCPL mutations disrupt the canonical CLV3/WUS feedback loop, uncovering their roles in SAM homeostasis. Additionally, knockout of BnaLFY homologs caused permanent vegetative state and sterility, demonstrating their conserved role in floral meristem identity in Brassica napus. Collectively, our study not only elucidates the critical function of BnaSCPLs in SAM maintenance but also establishes a regulatory framework for understanding meristem phase transitions in B. napus, providing potential targets for crop architecture improvement.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"55 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146070010","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}
SHOOT-MERISTEMLESS defines and serves as a marker for productive shoot progenitor cells, facilitating de novo shoot regeneration from callus in tissue culture.
{"title":"Stm is required for fate establishment of productive shoot progenitor cells in Arabidopsis tissue culture","authors":"Ning Zhai, Dixiang Xie, Lin Xu","doi":"10.1093/plphys/kiaf671","DOIUrl":"https://doi.org/10.1093/plphys/kiaf671","url":null,"abstract":"SHOOT-MERISTEMLESS defines and serves as a marker for productive shoot progenitor cells, facilitating de novo shoot regeneration from callus in tissue culture.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"7 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146071708","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}
Plant grafting, a series of tissue reunion processes, exhibits varying levels of compatibility across different species. Despite extensive research, the response of the scion to various compatible rootstocks remains poorly understood. In this study, we utilized transcriptomic analyses and gene functional validation experiments to investigate the role of xyloglucan endotransglucosylase/hydrolase (XTH) genes and their products in graft healing, specifically examining their effects on callus proliferation and graft survival in response to rootstocks with differing compatibilities across multiple species. Our results indicated that the less compatible bottle gourd rootstocks stimulate increased callus proliferation at the graft junctions with melon (Cucumis melo) scions. Virus-induced gene silencing of a highly expressed XTH gene, CmXTH9, in melon led to lower survival rates and reduced callus proliferation at the graft boundary. Furthermore, grafting accompanied upregulation of 25-55% of XTH family genes in the grafts of Arabidopsis thaliana, Nicotiana benthamiana, and Oryza sativa, which are distributed across different phylogenetic branches. Successful heterografts typically induced more family genes with greater upregulation than unsuccessful grafts. Consistently, an Arabidopsis Atxth4;Atxth7 mutant decreased grafting success rates and diminished callus proliferation at the wound site. These results underscore the conserved function of XTHs in graft union development and highlight their role in graft healing.
{"title":"Xyloglucan endotransglucosylase/hydrolase family genes are required for the plant graft union formation through callus proliferation.","authors":"Mu Xiong,Ting Zhang,Xin Qian,Akebaierjiang Kadeer,Ken-Ichi Kurotani,Ling Li,Changjin Liu,Xiangshuai Wu,Zhilong Bie,Michitaka Notaguchi,Yuan Huang","doi":"10.1093/plphys/kiag030","DOIUrl":"https://doi.org/10.1093/plphys/kiag030","url":null,"abstract":"Plant grafting, a series of tissue reunion processes, exhibits varying levels of compatibility across different species. Despite extensive research, the response of the scion to various compatible rootstocks remains poorly understood. In this study, we utilized transcriptomic analyses and gene functional validation experiments to investigate the role of xyloglucan endotransglucosylase/hydrolase (XTH) genes and their products in graft healing, specifically examining their effects on callus proliferation and graft survival in response to rootstocks with differing compatibilities across multiple species. Our results indicated that the less compatible bottle gourd rootstocks stimulate increased callus proliferation at the graft junctions with melon (Cucumis melo) scions. Virus-induced gene silencing of a highly expressed XTH gene, CmXTH9, in melon led to lower survival rates and reduced callus proliferation at the graft boundary. Furthermore, grafting accompanied upregulation of 25-55% of XTH family genes in the grafts of Arabidopsis thaliana, Nicotiana benthamiana, and Oryza sativa, which are distributed across different phylogenetic branches. Successful heterografts typically induced more family genes with greater upregulation than unsuccessful grafts. Consistently, an Arabidopsis Atxth4;Atxth7 mutant decreased grafting success rates and diminished callus proliferation at the wound site. These results underscore the conserved function of XTHs in graft union development and highlight their role in graft healing.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"1 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146073184","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}
Ultraviolet-B (UV-B) irradiation is an effective elicitor for the biosynthesis of pharmaceutically relevant monoterpenoid indole alkaloids (MIAs). Uncaria rhynchophylla (UR) produces numerous valuable MIAs, such as rhynchophylline and isorhynchophylline, which have huge chemotherapeutic potential. However, the mechanism underlying UV-B-induced MIAs remains elusive in MIA-producing plants. Here, we performed integrative transcriptome and metabolome analyses and found that UV-B distinctly induced MIA accumulation in UR leaves. Furthermore, we report that two UV-B-responsive transcription factors, UrWRKY40 and ELONGATED HYPOCOTYL 5 (UrHY5), cooperatively promote UV-B-induced MIA biosynthesis by activating MIA structural genes (UrSTR1, UrCPR1) via binding to their promoters (W-box and T/G-box elements). Comparative interactomics and dual-luciferase assays demonstrated that UrWRKY40 physically interacts with UrHY5 and represses its transcriptional activity under normal white light conditions. However, UV-B disrupted the formation of the UrWRKY40-UrHY5 complex and attenuated the repressive effects of UrWRKY40 on UrHY5 activity, thereby enhancing UrHY5-driven transactivation of downstream MIA structural genes. In addition, UV-B stimulated limited ABA production, which partially repressed UrWRKY40 expression, but not enough to override its induction by UV-B. In the presence of ABA, the UrWRKY40-UrHY5 interaction dissolved, which in turn released UrHY5 from repression, allowing it to activate MIA biosynthesis. These findings uncover a mechanism by which the UrWRKY40-UrHY5 module positively regulates UV-B-induced MIA biosynthesis by coordinating UV-B and ABA signaling, and provide a strategic framework for enhancing high-value MIA production through genetic manipulation.
{"title":"The UrWRKY40-UrHY5 module regul*ates the biosynthesis of UV-B-induced monoterpene indole alkaloids in Uncaria rhynchophylla.","authors":"Jia-Shun Yang,Hao-Cheng Lou,Hong Zhang,Xiao-Jun Pan,Xi Bao,Yu-Wen Qin,Yang-Ping Yang,Ren-Juan Qian,Pei-Long Wang,Jin-Guo Cheng,Zhi-Gang Wu","doi":"10.1093/plphys/kiag039","DOIUrl":"https://doi.org/10.1093/plphys/kiag039","url":null,"abstract":"Ultraviolet-B (UV-B) irradiation is an effective elicitor for the biosynthesis of pharmaceutically relevant monoterpenoid indole alkaloids (MIAs). Uncaria rhynchophylla (UR) produces numerous valuable MIAs, such as rhynchophylline and isorhynchophylline, which have huge chemotherapeutic potential. However, the mechanism underlying UV-B-induced MIAs remains elusive in MIA-producing plants. Here, we performed integrative transcriptome and metabolome analyses and found that UV-B distinctly induced MIA accumulation in UR leaves. Furthermore, we report that two UV-B-responsive transcription factors, UrWRKY40 and ELONGATED HYPOCOTYL 5 (UrHY5), cooperatively promote UV-B-induced MIA biosynthesis by activating MIA structural genes (UrSTR1, UrCPR1) via binding to their promoters (W-box and T/G-box elements). Comparative interactomics and dual-luciferase assays demonstrated that UrWRKY40 physically interacts with UrHY5 and represses its transcriptional activity under normal white light conditions. However, UV-B disrupted the formation of the UrWRKY40-UrHY5 complex and attenuated the repressive effects of UrWRKY40 on UrHY5 activity, thereby enhancing UrHY5-driven transactivation of downstream MIA structural genes. In addition, UV-B stimulated limited ABA production, which partially repressed UrWRKY40 expression, but not enough to override its induction by UV-B. In the presence of ABA, the UrWRKY40-UrHY5 interaction dissolved, which in turn released UrHY5 from repression, allowing it to activate MIA biosynthesis. These findings uncover a mechanism by which the UrWRKY40-UrHY5 module positively regulates UV-B-induced MIA biosynthesis by coordinating UV-B and ABA signaling, and provide a strategic framework for enhancing high-value MIA production through genetic manipulation.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"117 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146070011","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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