Plant Physiology and Biochemistry最新文献

{"title":"Identification of a nuclear transcription factor-Y (NF-Y) gene family in pepper and analysis of a CaNF-YC1-CaTIFY7-CaCBF1a/b transcription factor module under cold stress","authors":"Xiaodi Wang ,&nbsp;Kai Xu ,&nbsp;Ning Li ,&nbsp;Xiaohui Zhan ,&nbsp;Shenghua Gao ,&nbsp;Yanxu Yin ,&nbsp;Weiling Yuan ,&nbsp;Weifang Chen ,&nbsp;Minghua Yao ,&nbsp;Fei Wang","doi":"10.1016/j.plaphy.2025.110517","DOIUrl":"10.1016/j.plaphy.2025.110517","url":null,"abstract":"<div><div>Pepper is a popular crop, and its preference for a warm environment means that the frequent occurrence of extreme cold weather worldwide has increased the urgency to breed cold-tolerant vegetable varieties. The exploration of cold-tolerant genes has become a key research focus. However, research on transcription factors that regulate low-temperature tolerance in pepper plants remains limited. In this study, whole-genome analysis of the nuclear transcription factor Y (NF-Y) family was performed, revealing cold stress induction and transcription activation activity of <em>CaNF-YC1</em>. Silencing <em>CaNF-YC1</em> increased the H₂O₂ content and decreased the activities of the POD, SOD, and CAT enzymes under cold stress, significantly reducing the plants' low-temperature tolerance. Conversely, overexpression of <em>CaNF-YC1</em> enhanced pepper plants’ tolerance to low-temperature stress. RT-qPCR, yeast one-hybrid, and analysis of GUS/LUC reporter genes in tobacco demonstrated that the <em>CaNF-YC1</em> transcription factor activates expression of <em>CaCBF1a</em> and <em>CaCBF1b</em> by binding to their promoters<em>.</em> Yeast and tobacco assays revealed that the CaNF-YC1-CaTIFY7 complex finely regulates activation of the <em>CaCBF1a</em> and <em>CaCBF1b</em> promoters. The results indicate a crucial role for the CaNF-YC1-CaTIFY7-CaCBF1a/b molecular module in regulating core factors of low-temperature signaling in peppers.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110517"},"PeriodicalIF":5.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145207281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome-wide analysis of CNGC genes in grapevine (Vitis vinifera L.) and functional characterization of VvCNGC14 in silique development in Arabidopsis","authors":"Hongsheng Gao ,&nbsp;Hongying Wang ,&nbsp;BoHui Yu ,&nbsp;Xueting Wang ,&nbsp;Pan Guo ,&nbsp;Shiyang Li ,&nbsp;Jieshan Cheng ,&nbsp;Chunyan Yu ,&nbsp;Hongxia Zhang","doi":"10.1016/j.plaphy.2025.110526","DOIUrl":"10.1016/j.plaphy.2025.110526","url":null,"abstract":"<div><div>The cyclic nucleotide-gated channels (CNGCs) play a central role in calcium uptake. However, their accurate functions in the growth and response to biological stress in woody plants are still largely unknown. In this study, 22 <em>VvCNGC</em> genes were identified in the grapevine genome using BLAST and HMMER. Genome-wide association study (GWAS) analysis revealed a locus near VvCNGC14 associated with bunch weight and showing a suggestive trend with fertility. Further expression analysis showed that <em>VvCNGC14</em> was highly expressed in floral organ, particularly pollen. Heterologous expression of <em>VvCNGC14</em> in <em>Arabidopsis cngc18</em> mutant restored silique length, seed size, and seed weight to near wild-type levels, supporting a role in silique and seed development. Phylogenetic analysis revealed that <em>VvCNGC14</em> and <em>VvCNGC20</em> were closely related due to a segmental duplication event. Protein interaction network prediction identified a number of VvCNGC14-interacting proteins, including CDPK family members. GO analysis of predicted protein interaction highlighted enrichment in calcium-dependent protein kinase activity, protein autophosphorylation, adenyl ribonucleotide binding, and gibberellin 3-beta-dioxygenase activity. Expression analysis indicated strong pollen-specific expression of <em>VvCDPK7</em>, <em>VvCDPK8</em> and <em>VvCDPK16</em>, suggesting potential regulatory associations between <em>VvCNGC14</em> and these genes. Furthermore, salt and cold conditions elicited robust <em>VvCNGC2</em>/<em>7</em> induction (qRT-PCR), aligning with stress-responsive promoter motifs. Together, these findings highlight roles of CNGCs in development and abiotic-stress responses.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110526"},"PeriodicalIF":5.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145104816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcription factor PmNAC90 delays flower induction through IAA regulation in Prunus mume","authors":"Chengdong Ma ,&nbsp;Pengyu Zhou ,&nbsp;Yuying Lu ,&nbsp;Ximeng Lin ,&nbsp;Ziqi Wang ,&nbsp;Silas Segbo ,&nbsp;Kenneth Omondi Ouma ,&nbsp;Xiao Huang ,&nbsp;Ting Shi ,&nbsp;Zhaojun Ni ,&nbsp;Zhihong Gao","doi":"10.1016/j.plaphy.2025.110537","DOIUrl":"10.1016/j.plaphy.2025.110537","url":null,"abstract":"<div><div>NAC transcription factors play a pivotal role in plant development and stress responses, but their functional dynamics in flower induction remain unexplored in woody plant species. This study identified a NAC module, PmNAC90-PmNAC32, that controls flower induction in <em>Prunus mume</em>. Comprehensive interaction assays confirmed the direct physical binding between PmNAC90 and PmNAC32. Transcriptional profiling revealed that <em>PmNAC90</em> had significantly higher expression in late-flowering cultivar ‘Qingfeng’, and its expression was positively regulated by exogenous IAA treatment. <em>PmNAC90</em> overexpression in <em>Arabidopsis</em> significantly delayed the flowering time through transcriptional repression of core floral integrators and reduced endogenous IAA levels. Mechanistically, PmNAC90 directly activated transcription of the floral repressor <em>PmTFL1</em> and auxin glycosyltransferase <em>PmUGT84B</em>. PmNAC32 had antagonistic effects through dual regulatory mechanisms: transcriptional repression of <em>PmNAC90</em> expression and protein interaction-mediated inhibition of the transactivation activity of PmNAC90 on downstream targets. Our findings reveal a NAC-mediated regulatory switch that integrates protein interaction and hormonal signaling to fine-tune flower induction in perennial woody plants, providing critical insights into the evolutionary diversification of NAC functions in flowering control.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110537"},"PeriodicalIF":5.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aged and nano-sized polylactic acid enhanced arsenic accumulation and phytotoxicity in rice seedlings: Compared with micro-sized polylactic acid","authors":"Qiuying An ,&nbsp;Zhuo Zhen ,&nbsp;Fanyu Lin ,&nbsp;Changzhou Yan","doi":"10.1016/j.plaphy.2025.110535","DOIUrl":"10.1016/j.plaphy.2025.110535","url":null,"abstract":"<div><div>Rice is significantly impacted by the combined pollution of arsenic and plastic during cultivation, raising serious food safety concerns. However, the role of micro(nano)plastics in arsenic accumulation and the underlying toxic mechanisms remains unclear. Therefore, this research employed fluorescent labeling and laser denudation techniques to observe the in-situ distribution of polylactic acid (PLA) and arsenic in rice seedlings, aiming to elucidate the carrier impact of PLA on plant arsenic accumulation. It examined the effects of varying aged and sized PLA on total arsenic accumulation in plants, and investigated the effects of their combined pollution on the metabolic pathways of rice seedlings by untargeted metabolomics to elucidate potential toxic mechanism. The results indicated that PLA was an effective carrier, which co-transported with arsenic through xylem and accumulated in the veins of rice seedlings, thereby increasing total arsenic accumulation. Their combined pollution targeted the TCA cycle and phenylalanine biosynthesis as critical metabolic hubs, thus influenced downstream metabolites associated with oxidative stress resistance and impacted the growth of rice seedlings. This study confirmed the co-transport mechanism of PLA and arsenic via in-situ detection, which will enhance understanding of the mechanisms on which micro(nano)plastics increase the accumulation of heavy metals in plants and their health risks.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110535"},"PeriodicalIF":5.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrative LC-MS/MS and trait-based assessment of durum wheat genotypes: Insights into quality and stress tolerance from Algerian germplasm","authors":"Lamia Bourouh ,&nbsp;Nabila Souilah ,&nbsp;Mouad Boulacel ,&nbsp;Badis Aouzal ,&nbsp;Tahar Hazmoune ,&nbsp;Ibrahim Demirtas ,&nbsp;Ilyas Yildiz ,&nbsp;Fehmi Boufahja ,&nbsp;Hassan A. Rudayni ,&nbsp;Stefania Garzoli ,&nbsp;Hamdi Bendif","doi":"10.1016/j.plaphy.2025.110533","DOIUrl":"10.1016/j.plaphy.2025.110533","url":null,"abstract":"<div><div>Durum wheat (<em>Triticum durum</em> Desf.) constitutes a strategic cereal crop for Mediterranean agroecosystems, including Algeria. Enhancing its productivity and quality necessitates exploiting available genetic diversity. This investigation comprehensively evaluated eighteen durum wheat genotypes under subhumid field conditions in Skikda, Algeria, to quantify genetic variability through morpho-physiological traits, biochemical composition, and amino acid profiling via LC-MS/MS. Morpho-physiological assessments revealed disparities in germination capacity with <em>Ovidio</em> and <em>Cirta</em> showing 96 %, cold tolerance (min. 66.66 % in <em>Oued El Bared</em>), leaf relative water content (82.14 % in <em>Sigus</em>), and leaf area. Biochemical analyses demonstrated substantial ranges in wet gluten (26.64–34.52 %) and dry gluten (7.56–12.8 %), identifying <em>‘Cirta’</em> as superior for these quality determinants. Differential accumulation of osmolytes (soluble sugars, proline) and variations in photosynthetic pigments were observed. Phytochemical screening highlighted <em>‘El Tayeb’</em> for its markedly elevated total polyphenol concentration (28.17 mg GAE/g DE). High-resolution LC-MS/MS quantification of 16 amino acids delineated distinct compositional profiles, exemplified by high L-Alanine in <em>‘Oued El Bared</em>’ (1103.48 mg/100g) and L-Proline in <em>‘Bousselam’</em> (1333.44 mg/100g). This study quantifies significant genetic diversity within the evaluated germplasm, identifying genotypes with potentially superior agronomic adaptation, processing quality (e.g., <em>Cirta</em>), stress tolerance indicators (e.g., <em>Sigus</em> and <em>El Tayeb</em>), and nutritional profiles. These results provide a critical foundation for targeted germplasm utilization in Algerian durum wheat breeding programs aimed at improving yield stability, end-use quality, and resilience in subhumid environments.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110533"},"PeriodicalIF":5.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095166","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Coordinated regulation of enzymes and gene expression reveals thermotolerance mechanisms in Pleurotus giganteus","authors":"Yang Yang ,&nbsp;Frederick Leo Sossah ,&nbsp;Yongru Pian ,&nbsp;Zhu Lu ,&nbsp;Huan Wang ,&nbsp;Lianyan Bu ,&nbsp;Hanting Cheng ,&nbsp;Qinfen Li","doi":"10.1016/j.plaphy.2025.110532","DOIUrl":"10.1016/j.plaphy.2025.110532","url":null,"abstract":"<div><div>Thermotolerance is critical for the commercial cultivation of <em>Pleurotus giganteus</em> in tropical regions, especially under the impact of global climate change. This study investigated the thermotolerance mechanisms of two <em>P. giganteus</em> strains with contrasting heat responses: the heat-tolerant PG46 and heat-sensitive PG79. Phenotypic assessments, antioxidant and metabolic enzyme assays, and comparative transcriptome analyses were performed under heat stress at 40 °C. Both strains exhibited post-growth rings, reduced recovery, mycelial and membrane damage, which was more severe in PG79. Activities of antioxidant enzymes (superoxide dismutase, peroxidase, glutathione reductase, and catalase) and metabolic enzyme (laccase, cellulase, xylanase) initially increased and then declined, with PG46 consistently showing higher activity. Transcriptome analysis identified differentially expressed genes (DEGs) co-regulated in glycolysis, MAPK signaling, steroid biosynthesis, and protein processing pathways. In contrast, strain-specific expression patterns were observed in genes involved in ribonucleotide binding, zinc fingers, protein kinases, ubiquitination, cytochrome P450s, multicopper oxidases, MFS transporters, and 50S ribosome-binding GTPases, suggesting distinct molecular responses. Weighted Gene Co-expression Network Analysis (WGCNA) identified six gene modules associated with heat exposure duration and seven with thermotolerance. Key genes involved in signal transduction, protein processing, transferase activity, and macromolecule metabolism were identified. qRT-PCR validation of six selected genes confirmed RNA-seq reliability. These findings provided comprehensive insights into thermotolerance mechanisms in <em>P. giganteus</em>, offering specific gene targets and regulatory pathways for genetic improvement, precision breeding, and optimized cultivation strategies under heat stress conditions.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110532"},"PeriodicalIF":5.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ABA orchestrates molecular networks linking antioxidant defense, sugar metabolism and circadian clock for combined stress tolerance in Quinoa (Chenopodium quinoa Willd.)","authors":"Qiuwei Lu ,&nbsp;Shiting Bi ,&nbsp;Suxuan Dong ,&nbsp;Xueying Wang ,&nbsp;Fuye Guo ,&nbsp;Wenting Wang","doi":"10.1016/j.plaphy.2025.110536","DOIUrl":"10.1016/j.plaphy.2025.110536","url":null,"abstract":"<div><div>Quinoa (<em>Chenopodium quinoa</em> Willd.), an emerging model species for abiotic stress tolerance due to its exceptional environmental resilience, exhibits unique molecular adaptations to combined low temperature-drought stress. However, the mechanism by which ABA enhances combined stress tolerance in quinoa remains elusive. This study characterized the abscisic acid (ABA)-mediated regulatory network in quinoa using integrated physiological and transcriptomics analyses. We demonstrated that combined stress triggers ABA accumulation via significant up-regulating <em>NCED</em> - ABA biosynthesis genes (log₂(Fold Change) = 2.37 to 4.98, <em>p</em> &lt; 0.05) while suppressing <em>CYP707A</em> - ABA catabolism genes (log₂(Fold Change) = −1.81 to −2.99, <em>p</em> &lt; 0.05). Exogenous ABA application under low temperature-drought stress activated the PYR/PYL-PP2C-SnRK2 cascade, thereby enhancing antioxidant defenses (SOD, POD, CAT) and increasing the ratio of soluble sugars to starch, which mitigated oxidative damage. The ABA-responsive antioxidant module (MEwhite, 110 genes), identified via WGCNA, coordinates redox homeostasis and carbohydrate metabolism. This module relies on HSF-centered transcriptional networks to enhance stress tolerance. This module integrates ERF and WRKY transcription factors to coordinate stress adaptation, concurrently enhancing antioxidant enzyme activity for ROS scavenging while maintaining metabolic equilibrium under combinatorial stress. Crucially, circadian rhythm genes (<em>CRY</em>, <em>GI</em>, <em>PRR5</em>) coordinates both ABA-dependent and ABA-independent pathways enhance stress adaptation, revealing crosstalk between stress signaling and biological clocks. Fluridone-mediated ABA suppression exacerbated the inhibition of oxidoreductase activity and soluble sugar content, confirming ABA's pivotal role. These findings provide mechanistic insights into ABA-driven combinatorial stress adaptation, offering targets for breeding climate-resilient crops.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110536"},"PeriodicalIF":5.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Redirecting flavonoid flux in purple Chinese cabbage via Cas9-mediated BrDFR knockout","authors":"Sangkyu Park ,&nbsp;Hyo Lee ,&nbsp;Jaeeun Song ,&nbsp;Eun-Hye Kim ,&nbsp;Chan Ju Lim ,&nbsp;Jinpyo Oh ,&nbsp;Saet Buyl Lee ,&nbsp;Jin A. Kim ,&nbsp;Beom-Gi Kim","doi":"10.1016/j.plaphy.2025.110534","DOIUrl":"10.1016/j.plaphy.2025.110534","url":null,"abstract":"<div><div>Purple varieties of Chinese cabbage (<em>Brassica rapa</em> subsp. <em>pekinensis</em>) predominantly accumulate cyanidin-based anthocyanins. Although dihydroflavonol 4-reductase (DFR) is a key enzyme in anthocyanin biosynthesis, the function of the putative <em>B. rapa DFR</em> gene <em>(Bra027457</em>) remained unverified. We isolated and sequenced the coding region of <em>Bra027457</em> from four <em>B. rapa</em> inbred lines with either green or purple phenotypes and detected no sequence variation. <em>Bra027457</em> expression correlated with anthocyanin accumulation, and <em>in vitro</em> assays confirmed its ability to reduce all three dihydroflavonol substrates. Using CRISPR/Cas9, we knocked out <em>Bra027457</em> in the purple line 8267 and obtained transgene-free, homozygous <em>BrDFR</em>-KO plants. These exhibited a green phenotype due to complete anthocyanin loss, verifying <em>Bra027457</em> as the authentic <em>BrDFR</em> gene. Metabolite profiling of <em>BrDFR</em>-KO heads revealed significant increases in quercetin (Q), isorhamnetin (IR), and dihydroquercetin (DHQ). LC/MS analysis further identified five flavonol glycosides and one DHQ glycoside, of whch Q 3,7-di-<em>O</em>-glucoside and IR 3-<em>O</em>-(2‴-<em>O</em>-feruloyl)sophoroside-7-<em>O</em>-glucoside were predominant. These findings advance our understanding of flavonoid biosynthesis in <em>Brassica</em> species and provide valuable genetic resources for Chinese cabbage improvement.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110534"},"PeriodicalIF":5.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145095246","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment of a forward genetic screening system to discover new proteins associated with DNA demethylation pathway","authors":"Chong Lu ,&nbsp;Tiange Hu ,&nbsp;Jie Dai ,&nbsp;Yan Liu ,&nbsp;Yifei Sun ,&nbsp;Qianqian Wang ,&nbsp;Liran Sang ,&nbsp;Yi Zhou ,&nbsp;Mengqi Yang ,&nbsp;Yumei La ,&nbsp;Shaoxia Zhou ,&nbsp;Fei Chen ,&nbsp;Joseph Ndirangu Munyuru ,&nbsp;Honggui La","doi":"10.1016/j.plaphy.2025.110527","DOIUrl":"10.1016/j.plaphy.2025.110527","url":null,"abstract":"<div><div>Active DNA demethylation mediated by Repressor of Silencing 1 (ROS1) plays a major role in counteracting gene silencing and spreading of DNA methylation. However, the molecular mechanisms behind the ROS1-mediated DNA demethylation pathway remain largely unclear. To gain a deeper understanding of this process, we established a novel <em><u>H</u>TC<u>2</u>-p<u>U</u>bq10-Ω-Kozak</em>::<em><u>L</u>UC-1</em>-based (H2UL-1-based) forward genetic screening system to identify more protein factors involved in such a pathway. H2UL-1 transgenic line was created by introducing a <em>HTC2-pUbq10-Ω-Kozak</em>::<em>LUC</em> construct into Col-0 plants, and the H2UL-1 seedlings emitted strong luminescence in the presence of luciferase's substrate luciferin; however, introduction of a <em>ros1-4</em> mutation into the H2UL-1 background (designated H2UL-1/<em>ros1-4</em> line) caused marked transcriptional silencing of <em>Luciferase</em> (<em>LUC)</em> gene. A terminator sequence downstream of <em>LUC</em> (named <em>Ter</em>^LUC) was highly methylated in H2UL-1/<em>ros1-4</em> but not in H2UL-1 line, which may account for the silencing of <em>LUC</em> in the former. To test whether H2UL-1 line was suitable for identifying new candidate proteins, it was used as the starting material for mutant screening and subsequent gene cloning. Eventually, three genes were obtained, two of which were <em>DMETER-Like 2</em> (<em>DML2</em>) and <em>APEX1-LIKE</em> (<em>APE1L</em>) (which are well-known for involvement in the ROS1-mediated DNA demethylation pathway) and one of which was <em><u>R</u>epressor of <u>G</u>ene <u>S</u>ilencing 3</em> (<em>RGS3</em>), an SFMBT-like (SL) protein likely engaging in the modulation of chromatin conformation. Thus, the H2UL-1-based screening system appears to be effective in identifying candidate mutants (including new ones) and have the capacity to recover novel protein factors involved in ROS1-mediated DNA demethylation.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110527"},"PeriodicalIF":5.7,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145158006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"BZR1 coordinates multiple pathways to promote axillary bud outgrowth in model M. truncatula and forage yield in alfalfa","authors":"Yin Yanling,&nbsp;Liu Siyu,&nbsp;Wang Yumiao,&nbsp;Wu Jiarui,&nbsp;Xu Xiao,&nbsp;Fan Shugao","doi":"10.1016/j.plaphy.2025.110531","DOIUrl":"10.1016/j.plaphy.2025.110531","url":null,"abstract":"<div><div>Brassinosteroids (BRs) regulate plant architecture through the transcription factor BZR1, yet the molecular mechanisms underlying its role in axillary bud (AB) outgrowth and branching remain poorly characterized in <em>M</em>. <em>truncatula</em>. Here, we investigated the function of MtBZR1 using a null mutant (<em>mtbzr1</em>) through integrated phenotypic, transcriptomic, and hormonal analyses. Phenotypic comparisons revealed that the <em>mtbzr1</em> mutant exhibited a 22.9 % reduction in primary branches, shorter ABs, and significantly lower biomass accumulation than the wild type (WT). Treatment with 24-epibrassinolide (EBR) promoted AB elongation in isolated stem segments of WT, whereas this response was severely attenuated in <em>mtbzr1</em>, confirming the essential role of MtBZR1 in BR-mediated AB outgrowth. Transcriptomic profiling identified 27,624 differentially expressed genes (DEGs) in the ABs of <em>mtbzr1</em> compared to WT, with pronounced suppression of photosynthesis-related genes and cell cycle regulators. Hormone pathway analysis revealed transcriptional repression of auxin biosynthesis genes, coupled with elevated expression of strigolactone (SL) and abscisic acid (ABA) biosynthetic/signaling components. Quantification of endogenous hormones confirmed reduced indole-3-acetic acid (IAA) and elevated ABA levels in the ABs of <em>mtbzr1</em>. Overexpression of the homologous gene MsBZR1 (74.0 % protein identity to MtBZR1) in alfalfa (<em>Medicago sativa</em> L.) resulted in a 43.1–59.8 % increase in branching, consequently improving biomass, highlighting its yield promotion potential in forage crop. Collectively, these findings delineate the MtBZR1-mediated regulatory network controlling AB development in <em>M. truncatula</em> and provide strategic genetic targets for forage productivity enhancement.</div></div>","PeriodicalId":20234,"journal":{"name":"Plant Physiology and Biochemistry","volume":"229 ","pages":"Article 110531"},"PeriodicalIF":5.7,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145092310","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}